skip to Main Content

Low cost vibramycin

There’s a reason for that, too low cost vibramycin. For the past few weeks I’ve seen Facebook posts daily from former nursing colleagues in metro Detroit, one of the hardest hit areas in the country, as they provide front-line care to patients with COVID-19. It makes me very proud to call these nurses my friends. As a former emergency department low cost vibramycin nurse, I recall the feeling of satisfaction knowing that I’ve helped someone on the worst day of their life.

One of the best parts of being a nurse is knowing you matter to the only person in health care that truly matters. The patient. Several years ago I made the difficult decision to low cost vibramycin no longer perform bedside nursing and become a nurse administrator. The biggest loss from my transition is the feeling that what I do matters to the patient.

COVID-19 has forced a lot of us to rethink the role we play in health care and what the real priority should be. Things that were top priorities three months ago have been rightfully cast aside to either care for patients in low cost vibramycin a pandemic or prepare for the unknown future of, “When is our turn?. € For me, COVID-19 has reignited the feeling that what I do matters as virtual care has become a powerful tool on the forefront of care during this crisis. It has also shown that many of the powerful rules and regulations that limit virtual care are not needed and should be discarded permanently.

When I became the director of virtual care at our organization in 2015 I knew nothing about telehealth low cost vibramycin. Sure, I had seen a stroke robot in some Emergency Departments, and I had some friends that told me their insurance company lets them FaceTime a doctor for free (spoiler alert. It’s not FaceTime). I was tech-savvy from a consumer perspective and a tech novice from an IT low cost vibramycin perspective.

Nevertheless, my team and I spent the next few years learning as we built one of the higher volume virtual care networks in the state of Michigan. We discovered a lot of barriers that keep virtual care from actually making the lives of patients and providers better and we also became experts in working around those barriers. But, there were low cost vibramycin two obstacles that we could not overcome. Government regulation and insurance provider willingness to cover virtual visits.

These two barriers effectively cripple most legitimate attempts to provide value-added direct-to-consumer virtual care, which I define as using virtual care technologies to provide care outside of our brick-and-mortar facilities, most commonly in the patient home. The need to social distance, cancel appointments, close provider offices, keep from overloading emergency departments and urgent low cost vibramycin cares and shelter in place created instant demand for direct-to-consumer virtual care. In all honesty, I’ve always considered direct-to-consumer virtual care to be the flashy, must-have holiday gift of the year that organizations are convinced will be the way of the future. If a health system wants to provide on-demand access to patients for low-complexity acute conditions, they will easily find plenty of vendors that will sell them their app and their doctors and put the health system’s logo on it.

What a health system will struggle with is to find is enough patient demand low cost vibramycin to cover the high cost. Remember my friends from earlier that told me about the app their insurance gave them?. Nearly all of them followed that up by telling me they’ve never actually used it. I am fortunate that I low cost vibramycin work for an organization that understands this and instead focuses on how can we provide care that our patients actually want and need from the doctors they want to see.

Ironically, this fiscal year we had a corporate top priority around direct-to-consumer virtual care. We wanted to expand what we thought were some successful pilots and perform 500 direct-to-consumer visits. This year has been one of the hardest of my leadership career because, frankly, up until a month ago I was about to fail on this top priority low cost vibramycin. With only four months left, we were only about halfway there.

The biggest problem we ran into was that every great idea a physician brought to me was instantly dead in the water because practically no insurance company would pay for it. There are (prior to COVID-19) a plethora of rules around virtual care billing but the low cost vibramycin simplest way to summarize it is that most virtual care will only be paid if it happens in a rural location and inside of a health care facility. It is extremely limited what will be paid for in the patient home and most of it is so specific that the average patient isn’t eligible to get any in-home virtual care. Therefore, most good medical uses for direct-to-consumer care would be asking the patient to pay cash or the physician to forgo reimbursement for a visit that would be covered if it happened in office.

Add to that the massive capital and operating expenses it takes to build a virtual care network and you can low cost vibramycin see why these programs don’t exist. A month ago I was skeptical we’d have a robust direct-to-consumer program any time soon and then COVID-19 hit. When COVID-19 started to spread rapidly in the United States, regulations and reimbursement rules were being stripped daily. The first change that had major impact is when the Centers for Medicare and Medicaid Services (CMS) announced that they would temporarily begin reimbursing for virtual visits conducted in the patient’s home for COVID-19 and non-COVID related low cost vibramycin visits.

We were already frantically designing a virtual program to handle the wave of COVID-19 screening visits that were overloading our emergency departments and urgent cares. We were having plenty of discussions around reimbursement for this clinic. Do we attempt to bill insurances knowing they will likely deny, do we do a cash clinic model or do we do this as a community benefit and eat the low cost vibramycin cost?. The CMS waiver gave us hope that we would be compensated for diverting patients away from reimbursed visits to a virtual visit that is more convenient for the patient and aligns with the concept of social distancing.

Realistically we don’t know if we will be paid for any of this. We are holding all of the bills for low cost vibramycin at least 90 days while the industry sorts out the rules. I was excited by the reimbursement announcement because I knew we had eliminated one of the biggest direct-to-consumer virtual care barriers. However, I was quickly brought back to reality when I was reminded that HIPAA (Health Insurance Portability and Accountability Act) still existed.

I had this crazy idea that during a pandemic we should make it as easy as possible for people to receive virtual care and that the best way to do that was to meet the low cost vibramycin patient on the device they are most comfortable with and the application (FaceTime, Facebook, Skype, etc.) that they use every day. The problem is nearly every app the consumer uses on a daily basis is banned by HIPAA because “it’s not secure.” I’m not quite sure what a hacker stands to gain by listening into to my doctor and me talk about how my kids yet again gave me strep throat but apparently the concern is great enough to stifle the entire industry. Sure, not every health care discussion is as low-key as strep throat and a patient may want to protect certain topics from being discussed over a “non-secure” app but why not let the patient decide through informed consent?. Regulators could also abandon this all-or-nothing approach and lighten regulations low cost vibramycin surrounding specific health conditions.

The idea that regulations change based on medical situation is not new. For example, in my home state of Michigan, adolescents are essentially considered emancipated if it involves sexual health, mental health or substance abuse. Never mind that this same information is freely low cost vibramycin given over the phone by every office around the country daily without issue, but I digress. While my job is to innovate new pathways for care, our lawyer’s job is to protect the organization and he, along with IT security, rightfully shot down my consumer applications idea.

A few days later I legitimately screamed out loud in joy when the Department of Health and Human Services announced that it would use discretion on enforcing HIPAA compliance rules and specifically allowed for use of consumer applications. The elimination of billing restrictions and HIPAA regulations changed what is possible for health care organizations to low cost vibramycin offer virtually. Unfortunately both changes are listed as temporary and will likely be removed when the pandemic ends. Six days after the HIPAA changes were announced, we launched a centralized virtual clinic for any patient that wanted a direct-to-consumer video visit to be screened by a provider for COVID-19.

It allows patients to call in without a referral and most patients are on-screen within five minutes of low cost vibramycin clicking the link we text them. They don’t have to download an app, create an account or even be an established patient of our health system. It saw over 900 patients in the first 12 days it was open. That is 900 real patients low cost vibramycin that received care from a physician or advanced practice provider without risking personal exposure and without going to an already overwhelmed ED or urgent care.

To date, 70 percent of the patients seen by the virtual clinic did not meet CDC testing criteria for COVID-19. I don’t believe we could have reached even half of these patients had the consumer application restrictions been kept. A program like this almost certainly wouldn’t exist if not for the regulations being lifted and even low cost vibramycin if it did, it would have taken six to 12 months to navigate barriers and implement in normal times. Sure, the urgency of a pandemic helps but the impact of provider, patients, regulators and payors being on the same page is what fueled this fire.

During the virtual clinic’s first two weeks, my team turned its attention to getting over 300 providers across 60+ offices virtual so they could see their patients at home. Imagine being an immunocompromised cancer patient right now and being asked to leave your home and be exposed to other people in order to low cost vibramycin see your oncologist. Direct-to-consumer virtual care is the best way to safely care for these patients and without these temporary waivers it wouldn’t be covered by insurance even if you did navigate the clunky apps that are HIPAA compliant. Do we really think the immunocompromised cancer patient feels any more comfortable every normal flu season?.

Is it any more appropriate to low cost vibramycin ask them to risk exposure to the flu than it is to COVID-19?. And yet we deny them this access in normal times and it quite possibly will be stripped away from them when this crisis is over. Now 300 to 400 patients per day in our health system are seen virtually by their own primary care doctor or specialist for non-COVID related visits. Not a single one of these would have been reimbursed one month ago and I am highly skeptical I would have gotten approval to use the software low cost vibramycin that connects us to the patient.

Lastly, recall that prior to COVID-19, our system had only found 250 total patients that direct-to-consumer care was value-added and wasn’t restricted by regulation or reimbursement. COVID-19 has been a wake-up call to the whole country and health care is no exception. It has put priorities in perspective and shined low cost vibramycin a light on what is truly value-added. For direct-to-consumer virtual care it has shown us what is possible when we get out of our own way.

If a regulation has to be removed to allow for care during a crisis then we must question why it exists in the first place. HIPAA regulation cannot go back to its antiquated practices if we are truly going to shift the focus to patient low cost vibramycin wellness. CMS and private payors must embrace value-added direct-to-consumer virtual care and allow patients the access they deserve. COVID-19 has forced this industry forward, we cannot allow it to regress and be forgotten when this is over.

Tom Wood is the director of trauma and virtual care for MidMichigan Health, a low cost vibramycin non-profit health system headquartered in Midland, Michigan, affiliated with Michigan Medicine, the health care division of the University of Michigan. The views and opinions expressed in this commentary are his own.When dealing with all of the aspects of diabetes, it’s easy to let your feel fall to the bottom of the list. But daily care and evaluation is one of the best ways to prevent foot complications. It’s important to identify your risk factors and take the proper steps in low cost vibramycin limiting your complications.

Two of the biggest complications with diabetes are peripheral neuropathy and ulcer/amputation. Symptoms of peripheral neuropathy include numbness, tingling and/or burning in your feet and legs. You can slow the progression of developing neuropathy by making it a point to manage your blood sugars and keep them low cost vibramycin in the normal range. If you are experiencing these symptoms, it is important to establish and maintain a relationship with a podiatrist.

Your podiatrist can make sure things are looking healthy and bring things to your attention to monitor and keep a close eye on. Open wounds or ulcers can develop secondary to trauma, pressure, low cost vibramycin diabetes, neuropathy or poor circulation. If ulcerations do develop, it’s extremely important to identify the cause and address it. Ulcers can get worse quickly, so it’s necessary to seek immediate medical treatment if you find yourself or a loved one dealing with this complication.

Untreated ulcerations often lead to amputation and can be avoided if proper medical attention low cost vibramycin is sought right away. There are important things to remember when dealing with diabetic foot care. It’s very important to inspect your feet daily, especially if you have peripheral neuropathy. You may have a cut or a sore on your feet that you can’t feel, so your body doesn’t alarm you to check your feet.

Be gentle when bathing your feet. Moisturize your feet, but not between your toes. Do not treat calluses or corns on your own. Wear clean, dry socks.

Never walk barefoot, and consider socks and shoes made specifically for patients with diabetes. Kristin Raleigh, D.P.M., is a podiatrist who sees patients at Foot &.

How to get prescribed vibramycin

Vibramycin
Chloromycetin
Flagyl
Levaquin
Online price
Online
Online
Online
No
How long does stay in your system
RX pharmacy
RX pharmacy
At walmart
Nearby pharmacy
Dosage
At walmart
Drugstore on the corner
At walmart
At walmart
Buy with credit card
100mg 180 tablet $259.95
250mg 60 tablet $103.95
200mg 90 tablet $47.95
250mg 120 tablet $121.55
Without prescription
No
No
Yes
No
Best price for brand
Canadian pharmacy only
250mg
In online pharmacy
In online pharmacy

IntroductionThe lymphatic system is a network of vessels how to get prescribed vibramycin important for whole body fluid homeostasis, lipid absorption and immune cell trafficking.1 2 Lymphoedema is caused by lymphatic dysfunction, which leads to a build-up of interstitial fluid within the tissues. This manifests with swelling of the extremities, usually of the legs but may involve other regions or segments of the body such as the upper limbs, face, trunk or genital area. There is an increased risk of infection due to disturbances in immune cell trafficking within the segment of compromised lymph drainage.3 Lymphatic dysfunction within the thorax and abdomen, here referred to as systemic/internal involvement (but can be referred to as visceral or central involvement), may present with pleural or pericardial effusions or ascites, any of which may be chylous, as well as intestinal or pulmonary lymphangiectasia, protein losing enteropathy or chylous reflux.The International Society for the Study of Vascular Anomalies (ISSVA) updated their classification for vascular anomalies in 2018.4 The vascular malformations are subgrouped into ‘combined’, which include more than one type of vessel, ‘simple’ (only involving one vessel type), and those ‘associated with other anomalies’.Lymphoedema due to a presumed genetic developmental fault in the structure or function of lymph conducting pathways is called primary lymphoedema.5 Some developmental faults can lead to overt structural defects of the lymph conducting pathways and how to get prescribed vibramycin are called lymphatic malformations. Such malformations if interfering with lymph drainage cause lymphoedema (truncal malformations) but some lymphatic malformations remain as isolated anomalies with no connection to main lymph drainage pathways and do not cause lymphoedema (non-truncal malformations).6 A primary lymphatic anomaly is an umbrella term referring to all lymphatic abnormalities arising from a developmental fault.For a long time, the diagnosis of primary lymphoedema was based largely on the age of presentation of the swelling, congenital, pubertal and late onset, with limited differentiation between the phenotypes. The discovery of the first causal gene, vascular endothelial growth factor receptor 3 for Milroy disease, indicated that a molecular diagnosis was possible.7 The first St George’s how to get prescribed vibramycin classification algorithm of primary lymphoedema and other primary lymphatic disorders was an attempt to guide a clearer categorisation of phenotypes and enable the discovery of further causal genes.8 Age of onset remained a key criterion, but the sites affected and associated features, for example, dysmorphology, distichiasis (aberrant eyelashes), varicose veins, vascular malformations and limb overgrowth were also considered, as was internal or systemic involvement, for example, fetal hydrops, intestinal lymphangiectasia, pleural and pericardial effusions and chylous reflux.

A family history of lymphoedema with determination of the mode of inheritance was considered useful.More rigorous phenotyping facilitated the identification of subgroups of patients with the same broad category of primary lymphatic anomaly. These cohorts were then used for molecular studies to how to get prescribed vibramycin identify more causal genes. Once the genotype was known then crosschecking of the clinical characteristics, natural history and inheritance patterns was possible and an accurate phenotype defined. Investigations such as lymphoscintigraphy helped to refine the phenotype further how to get prescribed vibramycin and give insight into the mechanisms for the development of the lymphatic disorder. A first update of the classification was published in 2013.9The St George’s classification algorithm is intended to help clinicians categorise their patients and guide testing towards, where possible, a molecular diagnosis.

This algorithm is criteria matching, that is, using certain key findings for classification through a multistep process of history taking, how to get prescribed vibramycin examination findings, mutation testing, etc. The next step using the information gathered is to advise on natural history, prognosis and risk (including genetic counselling) and to guide management. While a molecular diagnosis should provide the most specific and accurate diagnosis, it can be seen particularly with the postzygotic mosaic disorders that one genotype can be clinically very heterogenous so there will probably always be a place for good clinical phenotyping supported by investigation to guide management.Here, we present a second update of the St George’s classification algorithm to include newly discovered genes and to bring it in-line with the 2018 ISSVA classification for vascular anomalies.4 The results of an audit, the purpose of which was to determine how well the algorithm was performing as a diagnostic aid to classify patients with primary lymphatic anomalies and guide molecular how to get prescribed vibramycin testing are also presented.MethodsSt George’s classification algorithm of primary lymphatic anomaliesThe St George’s classification algorithm was updated (figure 1) and then applied, retrospectively, to all patients presenting to the national multidisciplinary ‘Primary and Paediatric Lymphoedema’ Clinic held at St George’s Hospital over a 1-year period. Careful phenotyping was undertaken both on clinical grounds and after selective investigations, for example, lymphoscintigraphy. Where possible and appropriate, targeted genetic testing was performed (this how to get prescribed vibramycin was prior to the introduction of a lymphoedema gene panel in our unit) for some of the genes listed in table 1.St George’s classification algorithm for primary lymphatic anomalies.

The five main groupings (colour coded) with their various clinical subtypes of disease. Primary lymphoedema is the major clinical feature in how to get prescribed vibramycin the green, pink and purple sections. Text in red indicates the suggested genetic test and/or differential diagnosis for the subgroup, however, the indicated genes do not explain the cause of disease in all patients in each grouping. For example, only 70% how to get prescribed vibramycin of patients with Milroy disease are explained by mutations in FLT4/VEGFR3.33 FH, family history. +ve, positive.

ˆ’ve, negative how to get prescribed vibramycin. (Image shared by St George’s Lymphovascular Research Group under the CC BY-SA 4.0 International licence on Wikimedia Commons)." data-icon-position data-hide-link-title="0">Figure 1 St George’s classification algorithm for primary lymphatic anomalies. The five main groupings (colour coded) with their various clinical subtypes how to get prescribed vibramycin of disease. Primary lymphoedema is the major clinical feature in the green, pink and purple sections. Text in red indicates the suggested genetic test and/or differential diagnosis for the subgroup, however, the indicated genes do not explain the cause of disease in all patients in each grouping how to get prescribed vibramycin.

For example, only 70% of patients with Milroy disease are explained by mutations in FLT4/VEGFR3.33 FH, family history. +ve, positive how to get prescribed vibramycin. ˆ’ve, negative. (Image shared by St George’s Lymphovascular Research Group under the CC BY-SA 4.0 International licence on Wikimedia Commons).View this table:Table 1 An overview of genetic disorders with primary lymphoedema as a frequent and dominant feature, categorised by inheritance and age of onsetWithin the St George’s classification algorithm (figure 1), there are five main categories of primary lymphatic anomalies. These are presented how to get prescribed vibramycin in the form of colour-coded sections with the individual subtypes (including genotypes) within the categories.

For definitions of some of the terms used, see Glossary of Terms (see online supplementary section).Supplemental materialFirst, the yellow section includes the ‘vascular malformations associated with other anomalies’ and the ‘lymphatic malformations’ (as defined in the ‘Introduction’ section).Second, the patient is assessed for syndromes that have lymphoedema as a non-dominant feature (blue section), for example, the patient is dysmorphic with learning difficulties and possibly has other abnormalities.Then if not obviously syndromic, and the lymphatic problems are the dominant feature, further assessment and investigations for systemic/internal lymphatic dysfunction or central conducting anomalies (eg, chylothoraces, chylopericardial effusions, ascites or protein losing enteropathy) are undertaken (pink section). These include a careful how to get prescribed vibramycin medical history asking specifically about prenatal history (eg, hydrothoraces, fetal hydrops), chronic diarrhoea, abdominal bloating or discomfort with fatty foods, weight loss or faltering growth (in a child) or shortness of breath on exertion. Blood investigations (including serum albumin, immunoglobulins, lymphocyte subsets, faecal levels of calprotectin or alpha-1-antitrysin), echocardiograms and chest radiographs are helpful if central lymphatic dysfunction is suspected.Where none of the above features is present, then the age of onset is used to determine the grouping. The green section deals with congenital-onset primary lymphoedema (includes syndromes where lymphoedema is the dominant clinical problem, and which is present at birth or develops within the first year of life but is not associated with how to get prescribed vibramycin systemic/internal lymphatic dysfunction). The purple section addresses late-onset primary lymphoedema (ie, lymphoedema that is the dominant clinical problem, and which develops after the first year of life but is not associated with systemic/internal lymphatic dysfunction).

It was decided not to differentiate between pubertal onset (praecox) and later onset in how to get prescribed vibramycin life (tarda) when it was discovered that one genotype such as FOXC2 can cause both.It is important to note that the specific diagnosis may be difficult in a neonate presenting with isolated congenital primary lymphoedema. A baby born with lymphoedema may later present with developmental delay, systemic involvement, progressive segmental overgrowth or a vascular malformation, which could suggest a diagnosis in one of the other categories. It should how to get prescribed vibramycin also be emphasised that each colour-coded section is not exclusive. Some somatic overgrowth anomalies may possess significant internal involvement. Also, lymphoedema distichiasis syndrome is allocated to the purple late-onset lymphoedema how to get prescribed vibramycin section because the dominant feature is the late-onset lymphoedema not the associated features, which make it a syndrome.

The blue ‘syndromic’ section refers to conditions with a collection of features where lymphoedema is not the main characteristic. The algorithm how to get prescribed vibramycin is intended to guide a clinical diagnosis and target gene testing.Genetic methodologyFor the purposes of the audit, targeted genetic testing of FOXC2, VEGFR3, CCBE1, SOX18, RASopathy genes and PIK3CA was performed by Sanger sequencing of DNA extracted from lymphocytes or skin fibroblasts in patients in whom a specific genetic diagnosis was suspected. This was before the introduction of a lymphoedema gene panel. Some patients, who were either negative for the targeted genes or did not fit the relevant phenotypes of those genes, were included in Whole Exome Sequencing (WES) cohorts after classification, which then led to the identification of new disease genes such as EPHB4, GATA2, PIEZO1, GJC2 how to get prescribed vibramycin and FAT4.Retrospective audit of the St George’s Clinic for 2016A 12-month retrospective audit for the year 2016 (1 January 2016–31 December 2016) was performed. The aim of the audit was to look at the proportion of patients in each category of the classification algorithm and to look at the success of making a molecular diagnosis through use of the algorithm.

The audit criteria required the patients to be seen in our specialist clinic, at any age, with a diagnosis of a primary lymphatic anomaly with data collected from medical records and laboratory results.ResultsResults of the retrospective auditOver a 12-month period in 2016, 227 patients were seen how to get prescribed vibramycin (age range 2 weeks to 70 years), 25.6% (n=58/227) of which were new patients. Over one-third (38%) of patients seen in the clinic had a family history of primary lymphoedema.Few patients had received genetic testing prior to referral to the clinic. Targeted genetic testing was completed in 63% (n=143) of how to get prescribed vibramycin the patients seen. At that time, a lymphoedema gene panel was not available, patients were only tested if the clinician felt there was a reasonable chance of finding a molecular cause, that is, testing was targeted.Of those tested, the underlying genetic cause was identified in 41% (n=59/143). Overall, a how to get prescribed vibramycin molecular diagnosis was made in 26% (59/227) of all the patients seen in 2016.Vascular malformations with associated anomalies and lymphatic malformations (yellow)This group presents with malformations in the structure and organisation of blood and lymphatic vessels with a patchy, segmental distribution.

Lymphoedema may develop in combination with vascular malformations and segmental overgrowth (or occasionally, undergrowth) of tissues within the swollen limb, for example, muscle, skeletal or adipose tissues (figure 2A). The combination of lymphatic and vascular malformations in this group reflects the mutual embryological origins of the two vascular systems.A graphic representation of the 227 audited patients seen in clinic in 2016 and their distribution across the five categories from figure 1 how to get prescribed vibramycin (pie chart). (A–G) Images show features of each category. (A) Patients with postzygotic mutations often present with asymmetrical swelling and segmental overgrowth as this patient, who is mosaic for a mutation in KRAS how to get prescribed vibramycin. (B) Webbed neck in Noonan syndrome.

(C) In rare cases, swellings can be widespread affecting all segments of the body such as in this child with biallelic CCBE1 mutations. (D) In milder forms, often just the dorsum of the foot is affected how to get prescribed vibramycin as in this baby with a VEGFR3 mutation. (E, F) Lower limb swelling and distichiasis (arrowheads in F) in a patient with a FOXC2 mutation. (G) Lymphoedema is a major cause of skin disease and affected patients suffer from severe and recurrent episodes of cutaneous infection, especially HPV-associated warts as seen in patients with how to get prescribed vibramycin GATA2 mutations. GLD, generalised lymphatic dysplasia." data-icon-position data-hide-link-title="0">Figure 2 A graphic representation of the 227 audited patients seen in clinic in 2016 and their distribution across the five categories from figure 1 (pie chart).

(A–G) Images show features of how to get prescribed vibramycin each category. (A) Patients with postzygotic mutations often present with asymmetrical swelling and segmental overgrowth as this patient, who is mosaic for a mutation in KRAS. (B) Webbed neck in Noonan how to get prescribed vibramycin syndrome. (C) In rare cases, swellings can be widespread affecting all segments of the body such as in this child with biallelic CCBE1 mutations. (D) In milder forms, often just the dorsum of how to get prescribed vibramycin the foot is affected as in this baby with a VEGFR3 mutation.

(E, F) Lower limb swelling and distichiasis (arrowheads in F) in a patient with a FOXC2 mutation. (G) Lymphoedema is a major cause of skin disease and affected patients suffer from severe and recurrent episodes of cutaneous infection, especially HPV-associated warts as seen in patients with GATA2 mutations how to get prescribed vibramycin. GLD, generalised lymphatic dysplasia.These conditions are usually due to postzygotic mutations, for example, PIK3CA-related overgrowth spectrum (PROS)). Exceptions to this are capillary malformation-arteriovenous malformation (MIM 608354) such as Parkes-Weber syndrome, which may be caused by heterozygous, germline mutations how to get prescribed vibramycin in RASA1.10Of the 227 patients seen in 2016, 17% (n=39) had lymphoedema associated with vascular malformations and/or segmental overgrowth (or undergrowth) (figure 2, pie chart) in comparison with 15% in 2010.8 It has been shown that postzygotic, gain of function mutations in PIK3CA may be responsible for many of the mosaic segmental overgrowth spectrum disorders.11 Postzygotic mutations are rarely identified in blood samples and therefore require a skin biopsy of the affected region. In the 2016 cohort, only 10 patients (26%) provided skin biopsies for genetic analysis, producing just one molecular diagnosis.

More research in this field is required how to get prescribed vibramycin to identify the genetic basis for some of the conditions in this category. However, since the last revision, we have gained a much better understanding of the classification of some of these postzygotic mosaic conditions, therefore a brief review of the latest developments in this area is given in the online supplementary section.Syndromic lymphoedema (blue)Syndromes associated with primary lymphatic anomalies are listed in table 2 and include chromosomal abnormalities, single gene disorders and imprinting disorders. Patients attending the clinic with syndromic primary lymphoedema made up 13% (n=29) (figure 2, pie chart), similar to the 15% reported by Connell et al.8 Nearly three-quarters (72%, n=21) how to get prescribed vibramycin of this cohort had a molecular or chromosomal diagnosis. The most frequently seen syndromes were Noonan syndrome (n=8) (figure 2B), Turner syndrome (n=4) and Phelan McDermid syndrome (n=3).View this table:Table 2 An overview of ‘Known Syndromes’ with primary lymphoedema as a non-dominant association as referred to in the St George’s classification algorithm (figure 1, blue section)Lymphoedema with prenatal or postnatal systemic involvement (pink)In some conditions, lymphoedema may be associated with internal (systemic or visceral) disturbances of the lymphatic system within thorax or abdomen, for example, fetal hydrops, intestinal lymphangiectasia (presenting as protein-losing enteropathy), pulmonary lymphangiectasia or with pericardial and/or pleural effusions (often chylous), or chylous reflux (often into the genitalia). Broadly, there how to get prescribed vibramycin are two types of lymphoedema with systemic involvement.

(A) ‘widespread’ swelling affecting all segments of the body (figure 2C), such as that seen in generalised lymphatic dysplasia (GLD). Due to faulty development, the structural or functional abnormality of the lymphatic system is affecting the how to get prescribed vibramycin whole body. One type is Hennekam-lymphangiectasia-lymphoedema syndrome12. (B) ‘patchy’ areas of swelling, for example, left arm and right leg, which have been named ‘multisegmental how to get prescribed vibramycin lymphatic dysplasia’ (MLD) (figure 1).Prenatally, these conditions may present with pleural effusions (hydrothoraces), or as non-immune fetal hydrops (the accumulation of fluid in at least two compartments of a fetus such as the abdominal cavity, pleura or subcutaneous oedema). Fifteen per cent of non-immune cases of hydrops are the result of lymphatic disorders, and approximately 20% are idiopathic, some of which may be due to, as yet, unidentified lymphatic abnormalities.13In our audit, this cohort accounted for 12% (n=27) of patients (figure 2, pie chart), slightly higher than the 8% reported in 2010.8 Molecular testing was carried out in 17 patients.

Nine of those tested had GLD, and pathogenic variants were identified in seven (78%) how to get prescribed vibramycin. Five had biallelic variants in the PIEZO1 gene and one each with biallelic variants in FAT4 and SOX18. Interestingly, two of the families described by Connell et al, cases 3 and 4, have subsequently been found to be caused by biallelic variants in the PIEZO1 gene.8 14None of the eight patients, who presented with ‘patchy’ distribution of lymphoedema (MLD), had an identifiable molecular diagnosis. It is suspected that these patients could have a postzygotic mosaic mutation or WILD syndrome.15Since the last revision of the St George’s classification how to get prescribed vibramycin algorithm was published,9 five new causal genes associated with GLD and/or non-immune fetal hydrops have been identified. ADAMTS3,16 EPHB4,17 FAT4,18 FBXL719 and PIEZO114 20 and are reviewed in the online supplementary section.Congenital onset lymphoedema (green)In this category, congenital onset is defined as lymphoedema that is present at birth or develops within the first year of life.

Bilateral lower limb swelling is the most frequent presentation (figure how to get prescribed vibramycin 2D), but the swelling may be unilateral and/or involve the arms, genitalia and/or face, depending on the underlying cause. There are a number of different genetic disorders presenting with congenital lymphoedema (table 1). Milroy disease how to get prescribed vibramycin (ORPHA79452. OMIM 153100) is the most common form, occurring as a result of pathogenic variants in FLT4/VEGFR3.21 22 The mutation may occur de novo, so a family history is not essential for this diagnosis. The lymphoedema is always confined to how to get prescribed vibramycin the lower limbs but may be unilateral, and may (rarely) involve the genitalia.

Approximately 10% of mutation carriers do not have lymphoedema. Fetuses with Milroy disease may present antenatally with pedal oedema in the third trimester, and, in a few cases, with bilateral hydrothoraces, which resolve before birth.Pathogenic variants in VEGFC, the ligand for VEGFR3, have also been identified in association with congenital primary lymphoedema of Gordon (OMIM 615907), also affecting the lower limbs.23–26The congenital category how to get prescribed vibramycin represents 21% (n=47) of the patients seen in 2016 (figure 2, pie chart) compared with 24% in 2010.8 A pathogenic variant was identified in 19 of the 47 (40%) patients genetically tested in this category. The majority (n=18) had pathogenic variants identified in FLT4/VEGFR3 and, in one patient, a pathogenic variant in the GJC2 gene. A GJC2 mutation in a patient presenting with lymphoedema at birth is unusual but shows the variability of the phenotype.Many of the conditions listed under the other categories in the how to get prescribed vibramycin classification algorithm may initially present with congenital lymphoedema but systemic involvement, progressive overgrowth or vascular malformation may present later and are so reclassified. Likewise, some syndromic forms may present with congenital lymphoedema before any other manifestations, making diagnosis difficult at times.

Thus, the diagnosis of ‘isolated’ congenital primary how to get prescribed vibramycin lymphoedema may be difficult in a neonate presenting with pedal oedema. Therefore, a molecular diagnosis in the neonatal period is clinically very useful in the management of these patients.Late-onset lymphoedema (purple)‘Late-onset’ lymphoedema is defined as presenting after the first year of life. Swelling can range from being unilateral, bilateral or can involve all four limbs and how to get prescribed vibramycin can present from early childhood up to adulthood (figures 1 and 2E). Some may present with unilateral swelling, but the contralateral limb may become involved later or show abnormalities on lymphoscintigram even when clinically uninvolved. The phenotypes also range from mild to how to get prescribed vibramycin severe.

There are currently five genes known to be associated with late-onset lymphoedema. FOXC2 (figure 2F),27 GJC2,28 29 GATA2 how to get prescribed vibramycin (figure 2G),30 HGF31 and CELSR132 (table 1). For many patients the molecular cause remains elusive, particularly in those patients with Meige disease and late-onset (usually pubertal) unilateral lower limb lymphoedema.Late-onset primary lymphoedema accounted for 37% (n=85) in 2016 (figure 2, pie chart) comparable to the 36% reported in 2010.8 This category has a low number of molecular diagnoses (n=12. 14%) as there are currently no causative genes for Meige disease, which made up 36% (n=31) of patients in this category.DiscussionThis review presents an updated St George’s classification algorithm of primary lymphatic anomalies and brings it in-line with the how to get prescribed vibramycin ISSVA classification for vascular anomalies. It cites eight new causative genes since the last publication and highlights the areas where the genetic basis is still not known.

This rapidly evolving field demonstrates that primary lymphoedema and vascular malformations are highly heterogenous.The audit reports an overall successful molecular diagnosis in 26% of patients seen in the clinic, but 41% of those how to get prescribed vibramycin patients selected for molecular testing. This is a considerable improvement on the rate of a molecular diagnosis since the algorithm was first published in 2010. Only two how to get prescribed vibramycin causal genes were known at that time. We can conclude from the audit that the algorithm works well in targeting mutation testing. Furthermore, use of the algorithm has led to the discovery of a number of causal genes.

While it could be argued that the introduction of the lymphoedema gene panel obviates any need for targeted gene tests, we believe that matching a phenotype to a likely gene reduces wasteful testing and helps enormously in the interpretation of variants of unknown significance, which are becoming an increasing problem in the era of next-generation sequencing.Although providing a molecular diagnosis in one-quarter of how to get prescribed vibramycin all the patients with primary lymphoedema represents a considerable improvement from when the algorithm was last reviewed, the molecular diagnosis is still not identified in the majority of patients seen in the St George’s Clinic. In the diagnostic setting, the introduction of next-generation sequencing with a targeted (virtual) ‘lymphoedema gene panel’ may improve the diagnostic rate and broaden the phenotypic spectrum of many of the known genetic disorders. Understanding of the natural history of the disorder will enable appropriate surveillance of, for example, leukaemia in Emberger syndrome (GATA2), and allow investigations for how to get prescribed vibramycin known associated problems, for example, congenital heart disease in patients with lymphoedema distichiasis syndrome (FOXC2). Prenatal diagnosis for the more serious conditions also becomes possible. Knowledge of causal genes, and mechanisms how to get prescribed vibramycin of pathophysiology, provide an opportunity for new, improved treatments (personalised medicine) (eg, mammalian target of rapamycin inhibitors for progressive overgrowth disorders).In conclusion, the St George’s classification algorithm for primary lymphatic anomalies has been further refined.

With this review, we have provided insight into the most recently discovered genotypes and how this algorithm can be used in the clinic to guide management of patients with primary lymphoedema.IntroductionTriphalangeal thumb (TPT) is a rare congenital hand anomaly in which the thumb has three phalanges instead of two. TPT is usually inherited in an autosomal dominant trait and how to get prescribed vibramycin is therefore commonly seen in affected families. In 1994, Heutink et al located the pathogenic locus of TPT at chromosome 7q36.1 Subsequently, Lettice et al determined that point mutations in the zone of polarising activity regulatory sequence (ZRS) causes TPT and preaxial polydactyly.2 The ZRS is a long-range regulatory element residing in intron 5 of LMBR1 and regulates Sonic Hedgehog (SHH) expression in the embryonic limb bud. Since the identification of the ZRS region, 18 different point mutations in the ZRS have been reported in TPT how to get prescribed vibramycin families.3There is broad phenotypical variability among different point mutations in the ZRS. For example, variants on locations 323 and 739 in the ZRS cause mild presentations of isolated TPT.2 4 Alternatively, severe anomalies such as TPT accompanied with tibial hypoplasia have been observed in families with variants on position 404 and 406 in the ZRS.2 5–9 In mildly affected phenotypes, reduced penetrance is regularly observed.

In families who are more severely affected however, no reports of reduced penetrance have been made.Identifying and reporting new variants in the ZRS how to get prescribed vibramycin is important for genotype-phenotype correlations in TPT families. Additionally, it will also help to further elucidate the exact molecular mechanism of the role of the ZRS in the regulation of SHH expression in the embryonic limb.We therefore report two families with variants in the ZRS. These variants were identified in how to get prescribed vibramycin Dutch families with isolated TPT. Additionally, unaffected family members shared these variants with affected family members. Although this observation suggests that the genotype is not fully penetrant, how to get prescribed vibramycin minor anomalies within these presumed unaffected family members indicate subclinical expression of a TPT phenotype rather than reduced penetrance of the genotype.

We define subclinical phenotypes as anomalies that are not recognised by affected family members since they do not cause functional constraints in daily life, but can be recognised during clinical workup by experienced physicians.MethodsClinical evaluationFamilies 1 and 2 were identified at the outpatient clinic for Congenital Hand and Upper Limb Anomalies at the Sophia Children’s Hospital in Rotterdam, The Netherlands. The family members were clinically examined and how to get prescribed vibramycin consulted by a clinical geneticist. In family 1, peripheral blood samples were collected from the index patient, the mother and the grandfather of the index patient (figure 1). No blood samples were obtained from the brother of this patient as he was clinically unaffected and was below adult age.Overview of Dutch TPT how to get prescribed vibramycin family 1. (A) Pedigree of the Dutch TPT family 1.

The index how to get prescribed vibramycin patient is patient III-2. (B) X-ray image of the hand of the index patient. An additional deltaphalanx is present in both how to get prescribed vibramycin thumbs. (C) X-ray image of the thumbs of patient III-2. Although there is how to get prescribed vibramycin no triphalangism present, the thumbs are remarkably broad.

TPT, triphalangeal thumb." data-icon-position data-hide-link-title="0">Figure 1 Overview of Dutch TPT family 1. (A) Pedigree of the Dutch TPT family 1. The index how to get prescribed vibramycin patient is patient III-2. (B) X-ray image of the hand of the index patient. An additional deltaphalanx is present in both how to get prescribed vibramycin thumbs.

(C) X-ray image of the thumbs of patient III-2. Although there how to get prescribed vibramycin is no triphalangism present, the thumbs are remarkably broad. TPT, triphalangeal thumb.In family 2, the index patient (III-2) visited the outpatient clinic for Congenital Hand and Upper Limb Anomalies at the Sophia Children’s Hospital in Rotterdam with his parents. The other family members were how to get prescribed vibramycin visited as part of a field study. Included family members were clinically evaluated by a clinical geneticist, photographs were obtained and peripheral blood samples were collected (Figure 2, online supplementary figure 1).

No radiographs were obtained during the field study.Supplemental materialOverview of Dutch how to get prescribed vibramycin TPT family 2. (A) Outtake of pedigree of the Dutch TPT family 2. (B) Images of patient III-2 and his father (II-2), showing triphalangism of both thumbs how to get prescribed vibramycin with one additional ray on the left hand. (C) Images of patients II-4 and I-1, showing no triphalangism but lack of thumb opposition and mild thenar hypoplasia. TPT, triphalangeal thumb." data-icon-position data-hide-link-title="0">Figure 2 Overview of how to get prescribed vibramycin Dutch TPT family 2.

(A) Outtake of pedigree of the Dutch TPT family 2. (B) Images of patient III-2 how to get prescribed vibramycin and his father (II-2), showing triphalangism of both thumbs with one additional ray on the left hand. (C) Images of patients II-4 and I-1, showing no triphalangism but lack of thumb opposition and mild thenar hypoplasia. TPT, triphalangeal thumb.ZRS sequencingDNA samples were isolated from peripheral blood how to get prescribed vibramycin. The fragments were amplified using standard PCR.

An 834 bp fragment covering the ZRS how to get prescribed vibramycin (774 bp) was sequenced in family members of both families (UCSC Genome Browser, hg19, chr7:156583766–156584600). Sequencing of PCR products was executed using Big Dye Terminator 3.1. Fragments were loaded on an ABI 3130 Sequence analyser and genetic analysis was performed with SeqScape Software (V.3.0).ResultsClinical report​Family 1Family 1 (figure 1A) consists of a how to get prescribed vibramycin nuclear family containing two affected patients with TPT. The index patient had a bilateral isolated TPT with an additional deltaphalanx (figure 1B). No other congenital how to get prescribed vibramycin hand or other anomalies were present.

The mother of the index patient was born with a TPT accompanied with a rudimentary additional thumb on both hands, without any other hand or congenital anomaly (data not shown). The maternal grandfather of the index patient did not have a TPT how to get prescribed vibramycin or preaxial polydactyly. However, clinical examination of the hands revealed remarkable broadness of both thumbs and mild thenar hypoplasia. Although the X-ray image of the grandfather shows no duplication of the thumb or triphalangism, the broadness of the distal phalanges is striking (figure 1C).​Family 2Family 2 comprises a large seven-generation family (Figure 2A, online supplementary figure 1). The index patient (III-2) had bilateral TPT with preaxial how to get prescribed vibramycin polydactyly on the left hand.

The father of the index patient (II-1) had bilateral TPT without preaxial polydactyly (figure 2B). All other family members reported they were how to get prescribed vibramycin not affected. Although the thumbs of family members I-1 and II-2 did not show clear features of triphalangism, further examination revealed that both family members had mild thenar hypoplasia and were unable to oppose both thumbs (figure 2C). No other congenital anomalies were present in family 2.Mutation analysisSequence analysis of the 774 bp ZRS, in intron 5 of LMBR1, revealed the presence of a how to get prescribed vibramycin heterozygous A to G transition in members of family 1 (g.156584405A>G, GRCh37/Hg19). Following the more commonly used nomenclature for loci of ZRS variants, introduced by Lettice et al,2 this variant can be defined as a 165A>G variant.2 This variant was present in the affected family members.

Patient I-1 of family one also carried a 165A>G how to get prescribed vibramycin variant in the ZRS, despite not having TPT on either hand. This variant was not present in public databases dbSNP, Clinvar and HGMD. Additionally, this variant was not present in locally available WGS data sets (GoNL, Wellderly, Public54).10–12In family 2, we identified a 295T>variant how to get prescribed vibramycin in the ZRS (g.156584535T>C, GRCh37/Hg19). Two family members who did not have TPT carried the 295T>C variant. This variant has previously been reported in a British family with mild cases of TPT and reduced penetrance of how to get prescribed vibramycin the genotype.13 Additionally, transgenic enhancer assays in mice showed that the 295T>C variant causes ectopic expression in the embryonic limb and therefore confirms the pathogenicity of this variant.DiscussionIn this brief report, we describe two TPT families with either a 165A>G or 295T>C variant in the ZRS.

The aim of this paper was to show that these observations of reduced penetrance in TPT families are in retrospect caused by mild and subclinical limb phenotypes without the presence of triphalangism and therefore raise awareness for thorough clinical examination in members of TPT families who are presumed to be unaffected.Ever since the identification of ZRS by Lettice et al in 2003, 18 variants in ZRS have been published in the literature.2 4 6–9 13–20 These variants are generally fully penetrant and have been found in families with either TPT or TPT with preaxial polydactyly. Exceptions to the above how to get prescribed vibramycin are point mutations on positions 105, 404 and 406 in ZRS, which cause more severe phenotypes like tibial hypoplasia and polysyndactyly.2 5–9 21Although most variants in ZRS are considered fully penetrant, reduced penetrance has been reported in several TPT families with variants on positions 295, 334, 463 and 739 in ZRS.13 14 16 17The first aim of this paper is to hypothesise that some of these observations might not be caused by reduced penetrance of the genotype, but by a subclinical expression of the phenotype. We base our hypothesis on two arguments. First, family members who were initially presumed unaffected do show minor anomalies or altered hand function when examined how to get prescribed vibramycin appropriately. In family 1 of this study, the grandfather did not have TPT but had evident broadness of the thumb.

In family 2, patients with initially normal thumbs lacked the ability of opposition, which is caused by abnormal developmental patterning how to get prescribed vibramycin of the thumb. Although this observation is based on three patients from two families, we believe that these examples clearly illustrate our postulated hypothesis.Second, reports of non-penetrance are consistently associated with mild phenotypes in TPT families and not with severe TPT phenotypes, like tibial hypoplasia and polysyndactyly. This indicates that these observations only occur in TPT families where how to get prescribed vibramycin SHH expression is only slightly disrupted. In these families, the variability in the phenotypical spectrum is apparently broad enough that family members with variants in ZRS can present with subclinical phenotypes instead of TPT. However, it remains unclear why the disruption of SHH causes TPT in one family member how to get prescribed vibramycin and a subclinical phenotype in another.

One example of how intrafamilial variability can be explained is based on a reported family, where different degrees of somatic mosaicism were associated with various phenotypes in affected family members.22 As the regulatory function of ZRS on SHH is extremely delicate and affected by timing, location and level of activity, it is plausible that the slightest alteration of one of these factors can cause this interindividual phenotypical variation.The second aim of this paper is to underline the importance of two aspects when clinically examining and counselling patients with an inherited type of TPT. First, it is important to clinically investigate the presumed unaffected family how to get prescribed vibramycin members, as these patients might not encounter functional problems in their daily life and will report they are unaffected. However, a distinct broadness of the thumb, a double flexion fold in the thumb or a duplicated lunula might indicate a discrete inclination for duplication of the thumb or the presence of an additional phalanx. Additionally, functional limitations regarding thumb strength or lack of opposition should be evaluated as well how to get prescribed vibramycin. Second, presumed unaffected family members should only be informed that their future offspring have a population-wide probability of having TPT or polydactyly after genetic evaluation.

For complete reassurance, genetic evaluation of ZRS is also indicated for unaffected family members of mildly affected patients to verify whether they share the same disease-causing variant with their affected family members..

IntroductionThe lymphatic system is a network of vessels important for whole body fluid homeostasis, lipid absorption and immune cell trafficking.1 2 Lymphoedema is caused by lymphatic dysfunction, which low cost vibramycin leads to a build-up of interstitial fluid within the tissues. This manifests with swelling of the extremities, usually of the legs but may involve other regions or segments of the body such as the upper limbs, face, trunk or genital area. There is an increased risk of low cost vibramycin infection due to disturbances in immune cell trafficking within the segment of compromised lymph drainage.3 Lymphatic dysfunction within the thorax and abdomen, here referred to as systemic/internal involvement (but can be referred to as visceral or central involvement), may present with pleural or pericardial effusions or ascites, any of which may be chylous, as well as intestinal or pulmonary lymphangiectasia, protein losing enteropathy or chylous reflux.The International Society for the Study of Vascular Anomalies (ISSVA) updated their classification for vascular anomalies in 2018.4 The vascular malformations are subgrouped into ‘combined’, which include more than one type of vessel, ‘simple’ (only involving one vessel type), and those ‘associated with other anomalies’.Lymphoedema due to a presumed genetic developmental fault in the structure or function of lymph conducting pathways is called primary lymphoedema.5 Some developmental faults can lead to overt structural defects of the lymph conducting pathways and are called lymphatic malformations.

Such malformations if interfering with lymph drainage cause lymphoedema (truncal malformations) but some lymphatic malformations remain as isolated anomalies with no connection to main lymph drainage pathways and do not cause lymphoedema (non-truncal malformations).6 A primary lymphatic anomaly is an umbrella term referring to all lymphatic abnormalities arising from a developmental fault.For a long time, the diagnosis of primary lymphoedema was based largely on the age of presentation of the swelling, congenital, pubertal and late onset, with limited differentiation between the phenotypes. The discovery of the first causal gene, vascular endothelial growth factor receptor 3 for Milroy disease, indicated that a molecular diagnosis was possible.7 The first St George’s classification algorithm of primary low cost vibramycin lymphoedema and other primary lymphatic disorders was an attempt to guide a clearer categorisation of phenotypes and enable the discovery of further causal genes.8 Age of onset remained a key criterion, but the sites affected and associated features, for example, dysmorphology, distichiasis (aberrant eyelashes), varicose veins, vascular malformations and limb overgrowth were also considered, as was internal or systemic involvement, for example, fetal hydrops, intestinal lymphangiectasia, pleural and pericardial effusions and chylous reflux. A family history of lymphoedema with determination of the mode of inheritance was considered useful.More rigorous phenotyping facilitated the identification of subgroups of patients with the same broad category of primary lymphatic anomaly.

These cohorts were then used for molecular studies to identify more causal genes low cost vibramycin. Once the genotype was known then crosschecking of the clinical characteristics, natural history and inheritance patterns was possible and an accurate phenotype defined. Investigations such as lymphoscintigraphy helped to refine the phenotype further and give insight into the mechanisms for the development of the lymphatic disorder low cost vibramycin.

A first update of the classification was published in 2013.9The St George’s classification algorithm is intended to help clinicians categorise their patients and guide testing towards, where possible, a molecular diagnosis. This algorithm is criteria matching, that is, using low cost vibramycin certain key findings for classification through a multistep process of history taking, examination findings, mutation testing, etc. The next step using the information gathered is to advise on natural history, prognosis and risk (including genetic counselling) and to guide management.

While a molecular diagnosis should provide the most specific and accurate diagnosis, it can be seen particularly with the postzygotic mosaic disorders that one genotype can be clinically very heterogenous so there will probably always be a place for good clinical phenotyping supported by investigation to guide management.Here, we present a second update of the St George’s classification algorithm to include newly discovered genes and to bring it in-line with the 2018 ISSVA classification for vascular anomalies.4 The results of an audit, the low cost vibramycin purpose of which was to determine how well the algorithm was performing as a diagnostic aid to classify patients with primary lymphatic anomalies and guide molecular testing are also presented.MethodsSt George’s classification algorithm of primary lymphatic anomaliesThe St George’s classification algorithm was updated (figure 1) and then applied, retrospectively, to all patients presenting to the national multidisciplinary ‘Primary and Paediatric Lymphoedema’ Clinic held at St George’s Hospital over a 1-year period. Careful phenotyping was undertaken both on clinical grounds and after selective investigations, for example, lymphoscintigraphy. Where possible and appropriate, targeted genetic testing low cost vibramycin was performed (this was prior to the introduction of a lymphoedema gene panel in our unit) for some of the genes listed in table 1.St George’s classification algorithm for primary lymphatic anomalies.

The five main groupings (colour coded) with their various clinical subtypes of disease. Primary lymphoedema low cost vibramycin is the major clinical feature in the green, pink and purple sections. Text in red indicates the suggested genetic test and/or differential diagnosis for the subgroup, however, the indicated genes do not explain the cause of disease in all patients in each grouping.

For example, only 70% of patients with Milroy disease are explained by low cost vibramycin mutations in FLT4/VEGFR3.33 FH, family history. +ve, positive. ˆ’ve, negative low cost vibramycin.

(Image shared by St George’s Lymphovascular Research Group under the CC BY-SA 4.0 International licence on Wikimedia Commons)." data-icon-position data-hide-link-title="0">Figure 1 St George’s classification algorithm for primary lymphatic anomalies. The five main groupings (colour coded) low cost vibramycin with their various clinical subtypes of disease. Primary lymphoedema is the major clinical feature in the green, pink and purple sections.

Text in red indicates the suggested genetic test and/or differential diagnosis for the subgroup, however, the indicated genes do not explain the low cost vibramycin cause of disease in all patients in each grouping. For example, only 70% of patients with Milroy disease are explained by mutations in FLT4/VEGFR3.33 FH, family history. +ve, positive low cost vibramycin.

ˆ’ve, negative. (Image shared by St George’s Lymphovascular Research Group under the CC BY-SA 4.0 International licence on Wikimedia Commons).View this table:Table 1 An overview of genetic disorders with primary lymphoedema as a frequent and dominant feature, categorised by inheritance and age of onsetWithin the St George’s classification algorithm (figure 1), there are five main categories of primary lymphatic anomalies. These are presented in the form of colour-coded sections low cost vibramycin with the individual subtypes (including genotypes) within the categories.

For definitions of some of the terms used, see Glossary of Terms (see online supplementary section).Supplemental materialFirst, the yellow section includes the ‘vascular malformations associated with other anomalies’ and the ‘lymphatic malformations’ (as defined in the ‘Introduction’ section).Second, the patient is assessed for syndromes that have lymphoedema as a non-dominant feature (blue section), for example, the patient is dysmorphic with learning difficulties and possibly has other abnormalities.Then if not obviously syndromic, and the lymphatic problems are the dominant feature, further assessment and investigations for systemic/internal lymphatic dysfunction or central conducting anomalies (eg, chylothoraces, chylopericardial effusions, ascites or protein losing enteropathy) are undertaken (pink section). These include low cost vibramycin a careful medical history asking specifically about prenatal history (eg, hydrothoraces, fetal hydrops), chronic diarrhoea, abdominal bloating or discomfort with fatty foods, weight loss or faltering growth (in a child) or shortness of breath on exertion. Blood investigations (including serum albumin, immunoglobulins, lymphocyte subsets, faecal levels of calprotectin or alpha-1-antitrysin), echocardiograms and chest radiographs are helpful if central lymphatic dysfunction is suspected.Where none of the above features is present, then the age of onset is used to determine the grouping.

The green section deals with congenital-onset primary lymphoedema (includes syndromes where lymphoedema is the dominant low cost vibramycin clinical problem, and which is present at birth or develops within the first year of life but is not associated with systemic/internal lymphatic dysfunction). The purple section addresses late-onset primary lymphoedema (ie, lymphoedema that is the dominant clinical problem, and which develops after the first year of life but is not associated with systemic/internal lymphatic dysfunction). It was decided not to differentiate between pubertal low cost vibramycin onset (praecox) and later onset in life (tarda) when it was discovered that one genotype such as FOXC2 can cause both.It is important to note that the specific diagnosis may be difficult in a neonate presenting with isolated congenital primary lymphoedema.

A baby born with lymphoedema may later present with developmental delay, systemic involvement, progressive segmental overgrowth or a vascular malformation, which could suggest a diagnosis in one of the other categories. It should also be emphasised that each colour-coded low cost vibramycin section is not exclusive. Some somatic overgrowth anomalies may possess significant internal involvement.

Also, lymphoedema distichiasis syndrome is allocated to the purple late-onset lymphoedema section because the dominant feature is the late-onset lymphoedema not the associated features, which make it a low cost vibramycin syndrome. The blue ‘syndromic’ section refers to conditions with a collection of features where lymphoedema is not the main characteristic. The algorithm is intended to guide a clinical diagnosis and target gene testing.Genetic methodologyFor the purposes of the audit, targeted genetic testing of FOXC2, VEGFR3, CCBE1, SOX18, RASopathy genes and PIK3CA was performed by Sanger sequencing of DNA low cost vibramycin extracted from lymphocytes or skin fibroblasts in patients in whom a specific genetic diagnosis was suspected.

This was before the introduction of a lymphoedema gene panel. Some patients, who were either negative for the low cost vibramycin targeted genes or did not fit the relevant phenotypes of those genes, were included in Whole Exome Sequencing (WES) cohorts after classification, which then led to the identification of new disease genes such as EPHB4, GATA2, PIEZO1, GJC2 and FAT4.Retrospective audit of the St George’s Clinic for 2016A 12-month retrospective audit for the year 2016 (1 January 2016–31 December 2016) was performed. The aim of the audit was to look at the proportion of patients in each category of the classification algorithm and to look at the success of making a molecular diagnosis through use of the algorithm.

The audit criteria required the patients to be seen in our specialist clinic, at any age, with a low cost vibramycin diagnosis of a primary lymphatic anomaly with data collected from medical records and laboratory results.ResultsResults of the retrospective auditOver a 12-month period in 2016, 227 patients were seen (age range 2 weeks to 70 years), 25.6% (n=58/227) of which were new patients. Over one-third (38%) of patients seen in the clinic had a family history of primary lymphoedema.Few patients had received genetic testing prior to referral to the clinic. Targeted genetic testing was completed in low cost vibramycin 63% (n=143) of the patients seen.

At that time, a lymphoedema gene panel was not available, patients were only tested if the clinician felt there was a reasonable chance of finding a molecular cause, that is, testing was targeted.Of those tested, the underlying genetic cause was identified in 41% (n=59/143). Overall, a molecular diagnosis was made in 26% (59/227) of all the patients seen in 2016.Vascular malformations with associated anomalies and lymphatic malformations (yellow)This group presents with malformations in the structure and organisation low cost vibramycin of blood and lymphatic vessels with a patchy, segmental distribution. Lymphoedema may develop in combination with vascular malformations and segmental overgrowth (or occasionally, undergrowth) of tissues within the swollen limb, for example, muscle, skeletal or adipose tissues (figure 2A).

The combination low cost vibramycin of lymphatic and vascular malformations in this group reflects the mutual embryological origins of the two vascular systems.A graphic representation of the 227 audited patients seen in clinic in 2016 and their distribution across the five categories from figure 1 (pie chart). (A–G) Images show features of each category. (A) Patients with postzygotic mutations often present with asymmetrical swelling and segmental overgrowth as this patient, who is mosaic low cost vibramycin for a mutation in KRAS.

(B) Webbed neck in Noonan syndrome. (C) In rare cases, swellings can be widespread affecting all segments of the body such as in this child with biallelic CCBE1 mutations. (D) In milder forms, often just the dorsum of low cost vibramycin the foot is affected as in this baby with a VEGFR3 mutation.

(E, F) Lower limb swelling and distichiasis (arrowheads in F) in a patient with a FOXC2 mutation. (G) Lymphoedema is a major cause of low cost vibramycin skin disease and affected patients suffer from severe and recurrent episodes of cutaneous infection, especially HPV-associated warts as seen in patients with GATA2 mutations. GLD, generalised lymphatic dysplasia." data-icon-position data-hide-link-title="0">Figure 2 A graphic representation of the 227 audited patients seen in clinic in 2016 and their distribution across the five categories from figure 1 (pie chart).

(A–G) Images show low cost vibramycin features of each category. (A) Patients with postzygotic mutations often present with asymmetrical swelling and segmental overgrowth as this patient, who is mosaic for a mutation in KRAS. (B) Webbed neck in Noonan low cost vibramycin syndrome.

(C) In rare cases, swellings can be widespread affecting all segments of the body such as in this child with biallelic CCBE1 mutations. (D) In milder forms, often just the dorsum of the foot is affected as in this baby with a VEGFR3 low cost vibramycin mutation. (E, F) Lower limb swelling and distichiasis (arrowheads in F) in a patient with a FOXC2 mutation.

(G) Lymphoedema is a major cause of skin disease and affected patients suffer from severe and recurrent episodes of low cost vibramycin cutaneous infection, especially HPV-associated warts as seen in patients with GATA2 mutations. GLD, generalised lymphatic dysplasia.These conditions are usually due to postzygotic mutations, for example, PIK3CA-related overgrowth spectrum (PROS)). Exceptions to this are capillary malformation-arteriovenous malformation (MIM 608354) such as Parkes-Weber syndrome, which may be caused by heterozygous, germline mutations in RASA1.10Of the 227 patients seen in 2016, 17% (n=39) had lymphoedema associated with vascular malformations and/or segmental overgrowth (or undergrowth) (figure 2, pie chart) in comparison with 15% in 2010.8 It has been shown that postzygotic, gain of function mutations in PIK3CA may low cost vibramycin be responsible for many of the mosaic segmental overgrowth spectrum disorders.11 Postzygotic mutations are rarely identified in blood samples and therefore require a skin biopsy of the affected region.

In the 2016 cohort, only 10 patients (26%) provided skin biopsies for genetic analysis, producing just one molecular diagnosis. More research in this field is required to identify the genetic basis for some of low cost vibramycin the conditions in this category. However, since the last revision, we have gained a much better understanding of the classification of some of these postzygotic mosaic conditions, therefore a brief review of the latest developments in this area is given in the online supplementary section.Syndromic lymphoedema (blue)Syndromes associated with primary lymphatic anomalies are listed in table 2 and include chromosomal abnormalities, single gene disorders and imprinting disorders.

Patients attending the clinic low cost vibramycin with syndromic primary lymphoedema made up 13% (n=29) (figure 2, pie chart), similar to the 15% reported by Connell et al.8 Nearly three-quarters (72%, n=21) of this cohort had a molecular or chromosomal diagnosis. The most frequently seen syndromes were Noonan syndrome (n=8) (figure 2B), Turner syndrome (n=4) and Phelan McDermid syndrome (n=3).View this table:Table 2 An overview of ‘Known Syndromes’ with primary lymphoedema as a non-dominant association as referred to in the St George’s classification algorithm (figure 1, blue section)Lymphoedema with prenatal or postnatal systemic involvement (pink)In some conditions, lymphoedema may be associated with internal (systemic or visceral) disturbances of the lymphatic system within thorax or abdomen, for example, fetal hydrops, intestinal lymphangiectasia (presenting as protein-losing enteropathy), pulmonary lymphangiectasia or with pericardial and/or pleural effusions (often chylous), or chylous reflux (often into the genitalia). Broadly, there are two low cost vibramycin types of lymphoedema with systemic involvement.

(A) ‘widespread’ swelling affecting all segments of the body (figure 2C), such as that seen in generalised lymphatic dysplasia (GLD). Due to faulty development, the structural or functional abnormality of the low cost vibramycin lymphatic system is affecting the whole body. One type is Hennekam-lymphangiectasia-lymphoedema syndrome12.

(B) ‘patchy’ areas of swelling, for example, left low cost vibramycin arm and right leg, which have been named ‘multisegmental lymphatic dysplasia’ (MLD) (figure 1).Prenatally, these conditions may present with pleural effusions (hydrothoraces), or as non-immune fetal hydrops (the accumulation of fluid in at least two compartments of a fetus such as the abdominal cavity, pleura or subcutaneous oedema). Fifteen per cent of non-immune cases of hydrops are the result of lymphatic disorders, and approximately 20% are idiopathic, some of which may be due to, as yet, unidentified lymphatic abnormalities.13In our audit, this cohort accounted for 12% (n=27) of patients (figure 2, pie chart), slightly higher than the 8% reported in 2010.8 Molecular testing was carried out in 17 patients. Nine of those tested had GLD, low cost vibramycin and pathogenic variants were identified in seven (78%).

Five had biallelic variants in the PIEZO1 gene and one each with biallelic variants in FAT4 and SOX18. Interestingly, two of the families described by Connell et al, cases 3 and 4, have subsequently been found to be caused by biallelic variants in the PIEZO1 gene.8 14None of the eight patients, who presented with ‘patchy’ distribution of lymphoedema (MLD), had an identifiable molecular diagnosis. It is suspected that these patients could have a postzygotic mosaic mutation or WILD syndrome.15Since the last revision of the St George’s classification algorithm was published,9 five new causal genes associated low cost vibramycin with GLD and/or non-immune fetal hydrops have been identified.

ADAMTS3,16 EPHB4,17 FAT4,18 FBXL719 and PIEZO114 20 and are reviewed in the online supplementary section.Congenital onset lymphoedema (green)In this category, congenital onset is defined as lymphoedema that is present at birth or develops within the first year of life. Bilateral lower limb swelling is the most frequent presentation (figure 2D), but the swelling may low cost vibramycin be unilateral and/or involve the arms, genitalia and/or face, depending on the underlying cause. There are a number of different genetic disorders presenting with congenital lymphoedema (table 1).

Milroy disease low cost vibramycin (ORPHA79452. OMIM 153100) is the most common form, occurring as a result of pathogenic variants in FLT4/VEGFR3.21 22 The mutation may occur de novo, so a family history is not essential for this diagnosis. The lymphoedema is always confined to the lower limbs but may be unilateral, and may (rarely) involve the low cost vibramycin genitalia.

Approximately 10% of mutation carriers do not have lymphoedema. Fetuses with Milroy disease may present antenatally with pedal oedema in the third trimester, and, in a few cases, with bilateral hydrothoraces, which resolve before birth.Pathogenic variants in VEGFC, the ligand for VEGFR3, have also been identified in association with congenital primary lymphoedema of Gordon (OMIM 615907), also affecting the lower limbs.23–26The congenital category represents 21% (n=47) of the patients seen in 2016 (figure 2, pie chart) compared with 24% in 2010.8 A pathogenic variant was identified in 19 low cost vibramycin of the 47 (40%) patients genetically tested in this category. The majority (n=18) had pathogenic variants identified in FLT4/VEGFR3 and, in one patient, a pathogenic variant in the GJC2 gene.

A GJC2 mutation in a patient presenting with lymphoedema at low cost vibramycin birth is unusual but shows the variability of the phenotype.Many of the conditions listed under the other categories in the classification algorithm may initially present with congenital lymphoedema but systemic involvement, progressive overgrowth or vascular malformation may present later and are so reclassified. Likewise, some syndromic forms may present with congenital lymphoedema before any other manifestations, making diagnosis difficult at times. Thus, the diagnosis of ‘isolated’ congenital primary lymphoedema may be difficult in a neonate presenting with low cost vibramycin pedal oedema.

Therefore, a molecular diagnosis in the neonatal period is clinically very useful in the management of these patients.Late-onset lymphoedema (purple)‘Late-onset’ lymphoedema is defined as presenting after the first year of life. Swelling can range from being unilateral, bilateral or can involve all four limbs and low cost vibramycin can present from early childhood up to adulthood (figures 1 and 2E). Some may present with unilateral swelling, but the contralateral limb may become involved later or show abnormalities on lymphoscintigram even when clinically uninvolved.

The phenotypes also low cost vibramycin range from mild to severe. There are currently five genes known to be associated with late-onset lymphoedema. FOXC2 (figure 2F),27 GJC2,28 29 GATA2 (figure low cost vibramycin 2G),30 HGF31 and CELSR132 (table 1).

For many patients the molecular cause remains elusive, particularly in those patients with Meige disease and late-onset (usually pubertal) unilateral lower limb lymphoedema.Late-onset primary lymphoedema accounted for 37% (n=85) in 2016 (figure 2, pie chart) comparable to the 36% reported in 2010.8 This category has a low number of molecular diagnoses (n=12. 14%) as there are currently no causative genes for Meige disease, which made up 36% low cost vibramycin (n=31) of patients in this category.DiscussionThis review presents an updated St George’s classification algorithm of primary lymphatic anomalies and brings it in-line with the ISSVA classification for vascular anomalies. It cites eight new causative genes since the last publication and highlights the areas where the genetic basis is still not known.

This rapidly evolving field demonstrates that primary lymphoedema and vascular malformations are highly heterogenous.The audit reports an overall successful molecular diagnosis in 26% of patients seen in low cost vibramycin the clinic, but 41% of those patients selected for molecular testing. This is a considerable improvement on the rate of a molecular diagnosis since the algorithm was first published in 2010. Only two causal genes were known low cost vibramycin at that time.

We can conclude from the audit that the algorithm works well in targeting mutation testing. Furthermore, use of the algorithm has led to the discovery of a number of causal genes. While it could be argued that the introduction of the lymphoedema gene panel obviates any need for targeted gene tests, we believe that matching a phenotype to a likely gene reduces wasteful testing and helps enormously in the interpretation of variants of unknown significance, which are becoming an increasing problem in the era of next-generation sequencing.Although providing a molecular diagnosis in low cost vibramycin one-quarter of all the patients with primary lymphoedema represents a considerable improvement from when the algorithm was last reviewed, the molecular diagnosis is still not identified in the majority of patients seen in the St George’s Clinic.

In the diagnostic setting, the introduction of next-generation sequencing with a targeted (virtual) ‘lymphoedema gene panel’ may improve the diagnostic rate and broaden the phenotypic spectrum of many of the known genetic disorders. Understanding of the natural history of low cost vibramycin the disorder will enable appropriate surveillance of, for example, leukaemia in Emberger syndrome (GATA2), and allow investigations for known associated problems, for example, congenital heart disease in patients with lymphoedema distichiasis syndrome (FOXC2). Prenatal diagnosis for the more serious conditions also becomes possible.

Knowledge of causal genes, and mechanisms of pathophysiology, provide an opportunity for new, improved treatments (personalised medicine) low cost vibramycin (eg, mammalian target of rapamycin inhibitors for progressive overgrowth disorders).In conclusion, the St George’s classification algorithm for primary lymphatic anomalies has been further refined. With this review, we have provided insight into the most recently discovered genotypes and how this algorithm can be used in the clinic to guide management of patients with primary lymphoedema.IntroductionTriphalangeal thumb (TPT) is a rare congenital hand anomaly in which the thumb has three phalanges instead of two. TPT is usually inherited in an autosomal dominant trait and is therefore low cost vibramycin commonly seen in affected families.

In 1994, Heutink et al located the pathogenic locus of TPT at chromosome 7q36.1 Subsequently, Lettice et al determined that point mutations in the zone of polarising activity regulatory sequence (ZRS) causes TPT and preaxial polydactyly.2 The ZRS is a long-range regulatory element residing in intron 5 of LMBR1 and regulates Sonic Hedgehog (SHH) expression in the embryonic limb bud. Since the identification of the ZRS region, 18 different point mutations in the ZRS have been reported in TPT families.3There is broad phenotypical variability among different point mutations in low cost vibramycin the ZRS. For example, variants on locations 323 and 739 in the ZRS cause mild presentations of isolated TPT.2 4 Alternatively, severe anomalies such as TPT accompanied with tibial hypoplasia have been observed in families with variants on position 404 and 406 in the ZRS.2 5–9 In mildly affected phenotypes, reduced penetrance is regularly observed.

In families who are more severely affected however, no reports of reduced penetrance have been made.Identifying and reporting new variants in the ZRS is important low cost vibramycin for genotype-phenotype correlations in TPT families. Additionally, it will also help to further elucidate the exact molecular mechanism of the role of the ZRS in the regulation of SHH expression in the embryonic limb.We therefore report two families with variants in the ZRS. These variants were identified in Dutch families with isolated low cost vibramycin TPT.

Additionally, unaffected family members shared these variants with affected family members. Although this observation suggests that the genotype is not fully low cost vibramycin penetrant, minor anomalies within these presumed unaffected family members indicate subclinical expression of a TPT phenotype rather than reduced penetrance of the genotype. We define subclinical phenotypes as anomalies that are not recognised by affected family members since they do not cause functional constraints in daily life, but can be recognised during clinical workup by experienced physicians.MethodsClinical evaluationFamilies 1 and 2 were identified at the outpatient clinic for Congenital Hand and Upper Limb Anomalies at the Sophia Children’s Hospital in Rotterdam, The Netherlands.

The family members were clinically examined and low cost vibramycin consulted by a clinical geneticist. In family 1, peripheral blood samples were collected from the index patient, the mother and the grandfather of the index patient (figure 1). No blood samples were obtained from the brother of this patient as he was clinically unaffected and was below adult age.Overview low cost vibramycin of Dutch TPT family 1.

(A) Pedigree of the Dutch TPT family 1. The index patient is patient III-2 low cost vibramycin. (B) X-ray image of the hand of the index patient.

An additional deltaphalanx low cost vibramycin is present in both thumbs. (C) X-ray image of the thumbs of patient III-2. Although there is no triphalangism present, low cost vibramycin the thumbs are remarkably broad.

TPT, triphalangeal thumb." data-icon-position data-hide-link-title="0">Figure 1 Overview of Dutch TPT family 1. (A) Pedigree of the Dutch TPT family 1. The index patient is patient III-2 low cost vibramycin.

(B) X-ray image of the hand of the index patient. An additional deltaphalanx is present low cost vibramycin in both thumbs. (C) X-ray image of the thumbs of patient III-2.

Although there low cost vibramycin is no triphalangism present, the thumbs are remarkably broad. TPT, triphalangeal thumb.In family 2, the index patient (III-2) visited the outpatient clinic for Congenital Hand and Upper Limb Anomalies at the Sophia Children’s Hospital in Rotterdam with his parents. The other low cost vibramycin family members were visited as part of a field study.

Included family members were clinically evaluated by a clinical geneticist, photographs were obtained and peripheral blood samples were collected (Figure 2, online supplementary figure 1). No radiographs were obtained during the field study.Supplemental materialOverview of Dutch TPT family 2 low cost vibramycin. (A) Outtake of pedigree of the Dutch TPT family 2.

(B) Images of patient III-2 and his father (II-2), showing triphalangism of both thumbs with one additional ray on the low cost vibramycin left hand. (C) Images of patients II-4 and I-1, showing no triphalangism but lack of thumb opposition and mild thenar hypoplasia. TPT, triphalangeal thumb." data-icon-position data-hide-link-title="0">Figure 2 Overview of low cost vibramycin Dutch TPT family 2.

(A) Outtake of pedigree of the Dutch TPT family 2. (B) Images of patient III-2 and his father (II-2), showing triphalangism of low cost vibramycin both thumbs with one additional ray on the left hand. (C) Images of patients II-4 and I-1, showing no triphalangism but lack of thumb opposition and mild thenar hypoplasia.

TPT, triphalangeal thumb.ZRS sequencingDNA samples were isolated from low cost vibramycin peripheral blood. The fragments were amplified using standard PCR. An 834 bp fragment covering the ZRS (774 bp) was sequenced in family low cost vibramycin members of both families (UCSC Genome Browser, hg19, chr7:156583766–156584600).

Sequencing of PCR products was executed using Big Dye Terminator 3.1. Fragments were loaded on an ABI 3130 Sequence analyser and genetic analysis was performed with SeqScape Software (V.3.0).ResultsClinical report​Family 1Family 1 (figure 1A) consists low cost vibramycin of a nuclear family containing two affected patients with TPT. The index patient had a bilateral isolated TPT with an additional deltaphalanx (figure 1B).

No other congenital hand or other low cost vibramycin anomalies were present. The mother of the index patient was born with a TPT accompanied with a rudimentary additional thumb on both hands, without any other hand or congenital anomaly (data not shown). The maternal low cost vibramycin grandfather of the index patient did not have a TPT or preaxial polydactyly.

However, clinical examination of the hands revealed remarkable broadness of both thumbs and mild thenar hypoplasia. Although the X-ray image of the grandfather shows no duplication of the thumb or triphalangism, the broadness of the distal phalanges is striking (figure 1C).​Family 2Family 2 comprises a large seven-generation family (Figure 2A, online supplementary figure 1). The index low cost vibramycin patient (III-2) had bilateral TPT with preaxial polydactyly on the left hand.

The father of the index patient (II-1) had bilateral TPT without preaxial polydactyly (figure 2B). All other low cost vibramycin family members reported they were not affected. Although the thumbs of family members I-1 and II-2 did not show clear features of triphalangism, further examination revealed that both family members had mild thenar hypoplasia and were unable to oppose both thumbs (figure 2C).

No other congenital anomalies were present in family 2.Mutation analysisSequence analysis of the 774 bp ZRS, low cost vibramycin in intron 5 of LMBR1, revealed the presence of a heterozygous A to G transition in members of family 1 (g.156584405A>G, GRCh37/Hg19). Following the more commonly used nomenclature for loci of ZRS variants, introduced by Lettice et al,2 this variant can be defined as a 165A>G variant.2 This variant was present in the affected family members. Patient I-1 of family one also carried a 165A>G variant in the ZRS, despite not low cost vibramycin having TPT on either hand.

This variant was not present in public databases dbSNP, Clinvar and HGMD. Additionally, this variant was not present in locally available WGS data sets (GoNL, Wellderly, Public54).10–12In family 2, we identified a 295T>variant in low cost vibramycin the ZRS (g.156584535T>C, GRCh37/Hg19). Two family members who did not have TPT carried the 295T>C variant.

This variant low cost vibramycin has previously been reported in a British family with mild cases of TPT and reduced penetrance of the genotype.13 Additionally, transgenic enhancer assays in mice showed that the 295T>C variant causes ectopic expression in the embryonic limb and therefore confirms the pathogenicity of this variant.DiscussionIn this brief report, we describe two TPT families with either a 165A>G or 295T>C variant in the ZRS. The aim of this paper was to show that these observations of reduced penetrance in TPT families are in retrospect caused by mild and subclinical limb phenotypes without the presence of triphalangism and therefore raise awareness for thorough clinical examination in members of TPT families who are presumed to be unaffected.Ever since the identification of ZRS by Lettice et al in 2003, 18 variants in ZRS have been published in the literature.2 4 6–9 13–20 These variants are generally fully penetrant and have been found in families with either TPT or TPT with preaxial polydactyly. Exceptions to the above are point mutations on positions 105, 404 and 406 in ZRS, which cause more severe phenotypes like tibial hypoplasia and polysyndactyly.2 5–9 21Although most variants in ZRS are considered fully penetrant, reduced penetrance has been reported in several TPT families with variants on positions 295, 334, 463 and 739 in ZRS.13 14 16 17The first aim of this paper is to hypothesise that some of these observations might not be caused by reduced penetrance low cost vibramycin of the genotype, but by a subclinical expression of the phenotype.

We base our hypothesis on two arguments. First, family members who were low cost vibramycin initially presumed unaffected do show minor anomalies or altered hand function when examined appropriately. In family 1 of this study, the grandfather did not have TPT but had evident broadness of the thumb.

In family 2, patients with initially normal thumbs lacked the ability of opposition, which is caused low cost vibramycin by abnormal developmental patterning of the thumb. Although this observation is based on three patients from two families, we believe that these examples clearly illustrate our postulated hypothesis.Second, reports of non-penetrance are consistently associated with mild phenotypes in TPT families and not with severe TPT phenotypes, like tibial hypoplasia and polysyndactyly. This indicates that these observations only occur in TPT families where SHH expression is low cost vibramycin only slightly disrupted.

In these families, the variability in the phenotypical spectrum is apparently broad enough that family members with variants in ZRS can present with subclinical phenotypes instead of TPT. However, it low cost vibramycin remains unclear why the disruption of SHH causes TPT in one family member and a subclinical phenotype in another. One example of how intrafamilial variability can be explained is based on a reported family, where different degrees of somatic mosaicism were associated with various phenotypes in affected family members.22 As the regulatory function of ZRS on SHH is extremely delicate and affected by timing, location and level of activity, it is plausible that the slightest alteration of one of these factors can cause this interindividual phenotypical variation.The second aim of this paper is to underline the importance of two aspects when clinically examining and counselling patients with an inherited type of TPT.

First, it is important to clinically investigate the presumed unaffected family members, as these patients might low cost vibramycin not encounter functional problems in their daily life and will report they are unaffected. However, a distinct broadness of the thumb, a double flexion fold in the thumb or a duplicated lunula might indicate a discrete inclination for duplication of the thumb or the presence of an additional phalanx. Additionally, functional low cost vibramycin limitations regarding thumb strength or lack of opposition should be evaluated as well.

Second, presumed unaffected family members should only be informed that their future offspring have a population-wide probability of having TPT or polydactyly after genetic evaluation. For complete reassurance, genetic evaluation of ZRS is also indicated for unaffected family members of mildly affected patients to verify whether they share the same disease-causing variant with their affected family members..

How should I use Vibramycin?

Take Vibramycin by mouth with a full glass of water. Follow the directions on the prescription label. It is best to take Vibramycin without food, but if it upsets your stomach take it with food. Take your medicine at regular intervals. Do not take your medicine more often than directed. Take all of your medicine as directed even if you think you are better. Do not skip doses or stop your medicine early.

Talk to your pediatrician regarding the use of Vibramycin in children. Special care may be needed. While this drug may be prescribed for children as young as 8 years old for selected conditions, precautions do apply.

Overdosage: If you think you have taken too much of Vibramycin contact a poison control center or emergency room at once.

NOTE: Vibramycin is only for you. Do not share Vibramycin with others.

Buy vibramycin canada

IntroductionIn recent years, many studies have been published on new diagnostic possibilities and management approaches in cohorts of patients suspected to have a disorder/difference of sex development (DSD).1–13 Based on these studies, it has become clear that services and institutions still differ in the composition of the multidisciplinary teams buy vibramycin canada that provide care for patients who have a DSD.11 14 Several projects have now worked to resolve this variability in care. The European Cooperation in Science and Technology (EU COST) action BM1303 ‘A systematic elucidation of differences of sex development’ has been a platform to achieve European agreement on harmonisation of clinical management and laboratory practices.15–17 Another such initiative involved an update of the 2006 DSD consensus document by an international group of professionals and patient representatives.18 These initiatives have highlighted how cultural and financial aspects and the availability of resources differ significantly between countries and societies, a situation that hampers supranational agreement buy vibramycin canada on common diagnostic protocols. As only a few national guidelines have been published in international journals, comparison of these guidelines is difficult even though such a comparison is necessary to capture the differences and initiate actions to overcome them. Nonetheless, four DSD (expert) centres located in the Netherlands and Flanders (the Dutch-speaking Northern part of Belgium) have collaborated to produce buy vibramycin canada a detailed guideline on diagnostics in DSD.19 This shows that a supranational guideline can be a reasonable approach for countries with similarly structured healthcare systems and similar resources. Within the guideline there is agreement that optimisation of expertise and care can be achieved through centralisation, for example, by limiting analysis of next-generation sequencing (NGS)-based diagnostic panels to only a few centres and by centralising pathological review of gonadal tissues.

International networks such as the European Reference Network for rare endocrine conditions (EndoERN), in which DSD is embedded, may facilitate the expansion of this kind of collaboration across Europe.This paper highlights key discussion points in the buy vibramycin canada Dutch-Flemish guideline that have been insufficiently addressed in the literature thus far because they reflect evolving technologies or less visible stakeholders. For example, prenatal observation of an atypical aspect of the genitalia indicating a possible DSD is becoming increasingly common, and we discuss appropriate counselling and a diagnostic approach for these cases, including the option of using NGS-based genetic testing. So far, buy vibramycin canada little attention has been paid to this process.20 21 Furthermore, informing patients and/or their parents about atypical sex development and why this may warrant referral to a specialised team may be challenging, especially for professionals with limited experience in DSD.22 23 Therefore, a section of the Dutch-Flemish guideline was written for these healthcare providers. Moreover, this enables DSD specialists to refer to the guideline when advising a referral. Transition from the prenatal to the postnatal team and from the paediatric to the adult buy vibramycin canada team requires optimal communication between the specialists involved.

Application of NGS-based techniques may lead to a higher diagnostic yield, providing a molecular genetic diagnosis in previously unsolved cases.16 We address the timing of this testing and the problems associated with this technique such as the interpretation of variants of unknown clinical significance (VUS). Similarly, histopathological interpretation and classification of removed gonadal tissue is challenging and would benefit from international collaboration and centralisation of expertise.MethodsFor the guideline buy vibramycin canada revision, an interdisciplinary multicentre group was formed with all members responsible for updating the literature for a specific part of the guideline. Literature search in PubMed was not systematic, but rather intended to be broad in order to cover all areas and follow expert opinions. This approach is more in buy vibramycin canada line with the Clinical Practice Advisory Document method described by Burke et al24 for guidelines involving genetic practice because it is often troublesome to substantiate such guidelines with sufficient evidence due to the rapid changes in testing methods, for example, gene panels. All input provided by the group was synthesised by the chairperson (YvB), who also reviewed abstracts of papers on DSD published between 2010 and September 2017 for the guideline and up to October 2019 for this paper.

Abstracts had to be written in English and were identified using a broad range of Medical Subject Headings buy vibramycin canada terms (eg, DSD, genetic, review, diagnosis, diagnostics, 46,XX DSD, 46,XY DSD, guideline, multidisciplinary care). Next, potentially relevant papers on diagnostic procedures in DSD were selected. Case reports were excluded, as were buy vibramycin canada articles that were not open access or retrievable through institutional access. Based on this, a draft guideline was produced that was in line with the international principles of good diagnostic care in DSD. This draft was discussed by the writing committee and, after having obtained agreement on remaining buy vibramycin canada points of discussion, revised into a final draft.

This version was sent to a broad group of professionals from academic centres and DSD teams whose members had volunteered to review the draft guideline. After receiving and incorporating their input, buy vibramycin canada the final version was presented to the paediatric and genetic associations for approval. After approval by the members of the paediatric (NVK), clinical genetic (VKGN) and genetic laboratory (VKGL) associations, the guideline was published on their respective websites.19 Although Turner syndrome and Klinefelter syndrome are considered to be part of the DSD spectrum, they are not extensively discussed in this diagnostic guideline as guidelines dedicated to these syndromes already exist.25 26 However, some individuals with Turner syndrome or Klinefelter syndrome may present with ambiguous or atypical genitalia and may therefore initially follow the DSD diagnostic process.Guideline highlightsPrenatal settingPresentationThe most frequent prenatal presentation of a DSD condition is atypical genitalia found on prenatal ultrasound as an isolated finding or in combination with other structural anomalies. This usually occurs after the 20-week routine medical ultrasound for screening of congenital anomalies, but may also buy vibramycin canada occur earlier, for example, when a commercial ultrasound is performed at the request of the parents.Another way DSD can be diagnosed before birth is when invasive prenatal genetic testing carried out for a different reason, for example, due to suspicion of other structural anomalies, reveals a discrepancy between the genotypic sex and the phenotypic sex seen by ultrasound. In certified laboratories, the possibility of a sample switch is extremely low but should be ruled out immediately.

More often, the discrepancy will be due to sex-chromosome mosaicism or a true form of DSD.A situation now occurring with increasing frequency is a discrepancy between the genotypic sex revealed by non-invasive prenatal testing (NIPT), which is now available to high-risk pregnant women in the Netherlands and to all pregnant buy vibramycin canada women in Belgium, and later ultrasound findings. NIPT screens for CNVs in the fetus. However, depending on legal restrictions buy vibramycin canada and/or ethical considerations, the X and Y chromosomes are not always included in NIPT analysis and reports. If the X and Y chromosomes are included, it is important to realise that the presence of a Y-chromosome does not necessarily imply male fetal development. At the time that NIPT is performed (usually 11–13 weeks), genital development cannot be reliably appreciated by ultrasound, so any discrepancy or atypical aspect of the genitalia will only be noticed later in pregnancy and should prompt further evaluation.Counselling and diagnosticsIf a buy vibramycin canada DSD is suspected, first-line sonographers and obstetricians should refer the couple to their colleague prenatal specialists working with or in a DSD team.

After confirming an atypical genital on ultrasound, the specialist team should offer the couple a referral for genetic counselling to discuss the possibility of performing invasive prenatal testing (usually an amniocentesis) to identify an underlying cause that fits the ultrasound findings.22 23 To enable the parents to make a well-informed decision, prenatal counselling should, in our opinion, include. Information on the ultrasound findings and the limitations of buy vibramycin canada this technique. The procedure(s) that can be followed, including the risks associated with an amniocentesis. And the type of information genetic testing can and buy vibramycin canada cannot provide. Knowing which information has been provided and what words have been used by the prenatal specialist is very helpful for those involved in postnatal care.It is important that parents understand that the biological sex of a baby is determined by a complex interplay of chromosomes, genes and hormones, and thus that assessment of the presence or absence of a Y-chromosome alone is insufficient to assign the sex of their unborn child or, as in any unborn child, say anything about the child’s future gender identity.Expecting parents can be counselled by the clinical geneticist and the psychologist from the DSD team, although other DSD specialists can also be involved.

The clinical geneticist should be experienced in prenatal counselling and well informed about the diagnostic possibilities given the limited time span in which test results need to be buy vibramycin canada available to allow parents to make a well-informed decision about whether or not to continue the pregnancy. Termination of pregnancy can be buy vibramycin canada considered, for instance, in a syndromic form of DSD with multiple malformations, but when the DSD occurs as an apparently isolated condition, expecting parents may also consider termination of pregnancy, which, although considered controversial by some, is legal in Belgium and the Netherlands. The psychologist of the DSD team can support parents during and after pregnancy and help them cope with feelings of uncertainty and eventual considerations of a termination of pregnancy, as well as with practical issues, for example, how to inform others. The stress buy vibramycin canada of not knowing exactly what the child’s genitalia will look like and uncertainty about the diagnosis, treatment and prognosis cannot be avoided completely. Parents are informed that if the postnatal phenotype is different from what was prenatally expected, the advice given about diagnostic testing can be adjusted accordingly, for example, if a hypospadias is milder than was expected based on prenatal ultrasound images.

In our experience, parents appreciate having already spoken to some members of the DSD team during pregnancy and having a contact person before birth.After expert prenatal counselling, a significant number of pregnant buy vibramycin canada couples decline prenatal testing (personal experience IALG, MK, ABD, YvB, MC and HC-vdG). At birth, umbilical cord blood is a good source for (molecular) karyotyping and storage of DNA and can be obtained by the obstetrician, midwife or neonatologist. The terminology used in communication with parents should be carefully chosen,22 23 and midwives and staff of neonatal and delivery units should be clearly instructed to use gender-neutral and non-stigmatising vocabulary (eg, ‘your baby’) as long as sex assignment is pending.An algorithm for diagnostic evaluation of a suspected DSD in the prenatal situation is buy vibramycin canada proposed in figure 1. When couples opt for invasive prenatal diagnosis, the genetic analysis usually involves an (SNP)-array. It was recently estimated that >30% of individuals who have a DSD have additional structural anomalies, with cardiac and neurological anomalies and fetal growth restriction being particularly common.27 buy vibramycin canada 28 If additional anomalies are seen, the geneticist can consider specific gene defects that may underlie a known genetic syndrome or carry out NGS.

NGS-based techniques have also now made their appearance in prenatal diagnosis of congenital anomalies.29 30 Panels using these techniques can be specific for genes involved in DSD, or be larger panels covering multiple congenital anomalies, and are usually employed with trio-analysis to compare variants identified in the child with the parents’ genetics.29–31 Finding a genetic cause before delivery can help reduce parental stress in the neonatal period and speed up decisions regarding gender assignment. In such cases there is no tight time limit, and we propose completing the analysis well before the expected delivery.Disorders/differences of sex development buy vibramycin canada (DSD) in the prenatal setting. A diagnostic algorithm. *SOX9. Upstream anomalies and balanced translocations at promotor sites!.

Conventional karyotyping can be useful. NGS, next-generation sequencing." data-icon-position data-hide-link-title="0">Figure 1 Disorders/differences of sex development (DSD) in the prenatal setting. A diagnostic algorithm. *SOX9. Upstream anomalies and balanced translocations at promotor sites!.

Conventional karyotyping can be useful. NGS, next-generation sequencing.First contact by a professional less experienced in DSDWhereas most current guidelines start from the point when an individual has been referred to the DSD team,1 15 the Dutch-Flemish guideline dedicates a chapter to healthcare professionals less experienced in DSD as they are often the first to suspect or identify such a condition. Apart from the paper of Indyk,7 little guidance is available for these professionals about how to act in such a situation. The chapter in the Dutch-Flemish guideline summarises the various clinical presentations that a DSD can have and provides information on how to communicate with parents and/or patients about the findings of the physical examination, the first-line investigations and the need for prompt referral to a specialised centre for further evaluation. Clinical examples are offered to illustrate some of these recurring situations.

The medical issues in DSD can be very challenging, and the social and psychological impact is high. For neonates with ambiguous genitalia, sex assignment is an urgent and crucial issue, and it is mandatory that parents are informed that it is possible to postpone registration of their child’s sex. In cases where sex assignment has already taken place, the message that the development of the gonads or genitalia is still atypical is complicated and distressing for patients and parents or carers. A list of contact details for DSD centres and patient organisations in the Netherlands and Flanders is attached to the Dutch-Flemish guideline. Publishing such a list, either in guidelines or online, can help healthcare professionals find the nearest centres for consultations and provide patients and patient organisations with an overview of the centres where expertise is available.Timing and place of genetic testing using NGS-based gene panelsThe diagnostic workup that is proposed for 46,XX and 46,XY DSD is shown in figures 2 and 3, respectively.

Even with the rapidly expanding molecular possibilities, a (family) history and a physical examination remain the essential first steps in the diagnostic process. Biochemical and hormonal screening aim at investigating serum electrolytes, renal function and the hypothalamic-pituitary-gonadal and hypothalamic-pituitary-adrenal axes. Ultrasound screening of kidneys and internal genitalia, as well as establishing genotypic sex, should be accomplished within 48 hours and complete the baseline diagnostic work-up of a child born with ambiguous genitalia.1 16 32 3346,XX disorders/differences of sex development (DSD) in the postnatal setting. A diagnostic algorithm. NGS, next-generation sequencing.

CAH, Congenital adrenal hyperplasia. AMH, Anti-Müllerian Hormone." data-icon-position data-hide-link-title="0">Figure 2 46,XX disorders/differences of sex development (DSD) in the postnatal setting. A diagnostic algorithm. NGS, next-generation sequencing. CAH, Congenital adrenal hyperplasia.

AMH, Anti-Müllerian Hormone.46,XY disorders/differences of sex development (DSD) in the postnatal setting. A diagnostic algorithm. * SOX9. Upstream anomalies and balanced translocations at promotor sites!. Conventional karyotyping can be useful.

NGS, next-generation sequencing." data-icon-position data-hide-link-title="0">Figure 3 46,XY disorders/differences of sex development (DSD) in the postnatal setting. A diagnostic algorithm. *SOX9. Upstream anomalies and balanced translocations at promotor sites!. Conventional karyotyping can be useful.

NGS, next-generation sequencing.Very recently, a European position paper has been published focusing on the genetic workup of DSD.16 It highlights the limitations and drawbacks of NGS-based tests, which include the chance of missing subtle structural variants such as CNVs and mosaicism and the fact that NGS cannot detect methylation defects or other epigenetic changes.16 28 31 Targeted DNA analysis is preferred in cases where hormonal investigations suggest a block in steroidogenesis (eg, 11-β-hydroxylase deficiency, 21-hydroxylase deficiency), or in the context of a specific clinical constellation such as the often coincidental finding of Müllerian structures in a boy with normal external genitalia or cryptorchidism, that is, persistent Müllerian duct syndrome.33 34 Alternative tests should also be considered depending on the available information. Sometimes, a simple mouth swab for FISH analysis can detect mosaic XY/X in a male with hypospadias or asymmetric gonadal development or in a female with little or no Turner syndrome stigmata and a normal male molecular karyotyping profile or peripheral blood karyotype. Such targeted testing avoids incidental findings and is cheaper and faster than analysis of a large NGS-based panel, although the cost difference is rapidly declining.However, due to the genetic and phenotypic heterogeneity of DSD conditions, the most cost-effective next steps in the majority of cases are whole exome sequencing followed by panel analysis of genes involved in genital development and function or trio-analysis of a large gene panel (such as a Mendeliome).16 35–38 Pretest genetic counselling involves discussing what kind of information will be reported to patients or parents and the chance of detecting VUS, and the small risk of incidental findings when analysing a DSD panel should be mentioned. Laboratories also differ in what class of variants they report.39 In our experience, the fear of incidental findings is a major reason why some parents refrain from genetic testing.Timing of the DSD gene panel analysis is also important. While some patients or parents prefer that all diagnostic procedures be performed as soon as possible, others need time to reflect on the complex information related to more extensive genetic testing and on its possible consequences.

If parents or patients do not consent to panel-based genetic testing, analysis of specific genes, such as WT1, should be considered when appropriate in view of the clinical consequences if a mutation is present (eg, clinical surveillance of renal function and screening for Wilms’ tumour in the case of WT1 mutations). Genes that are more frequently involved in DSD (eg, SRY, NR5A1) and that match the specific clinical and hormonal features in a given patient could also be considered for sequencing. Targeted gene analysis may also be preferred in centres located in countries that do not have the resources or technical requirements to perform NGS panel-based genetic testing. Alternatively, participation by these centres in international collaborative networks may allow them to outsource the molecular genetic workup abroad.Gene panels differ between centres and are regularly updated based on scientific progress. A comparison of DSD gene panels used in recent studies can be found at https://www.nature.com/articles/s41574-018-0010-8%23Sec46.15 The panels currently used at the coauthors’ institutions can be found on their respective websites.

Given the pace of change, it is important to regularly consider repeating analysis in patients with an unexplained DSD, for example, when they transition into adult care or when they move from one centre to another. This also applies to patients in whom a clinical diagnosis has never been genetically confirmed. Confusion may arise when the diagnosis cannot be confirmed or when a mutation is identified in a different gene, for example, NR5A1 in someone with a clinical diagnosis of CAIS that has other consequences for relatives. Hence, new genetic counselling should always accompany new diagnostic endeavours.Class 3 variants and histopathological examinationsThe rapidly evolving diagnostic possibilities raise new questions. What do laboratories report?.

How should we deal with the frequent findings of mainly unique VUS or class 3 variants (ACMG recommendation) in the many different DSD-related genes in the diagnostic setting?. Reporting VUS can be a source of uncertainty for parents, but not reporting these variants precludes further investigations to determine their possible pathogenicity. It can also be difficult to prove variant pathogenicity, both on gene-level and variant-level.39 Moreover, given the gonad-specific expression of some genes and the variable phenotypic spectrum and reduced penetrance, segregation analysis is not always informative. A class 3 variant that does not fit the clinical presentation may be unrelated to the observed phenotype, but it could also represent a newly emerging phenotype. This was recently demonstrated by the identification of the NR5A1 mutation, R92W, in individuals with 46,XX testicular and ovotesticular DSD.40 This gene had previously been associated with 46,XY DSD.

In diagnostic laboratories, there is usually no capacity or expertise to conduct large-scale functional studies to determine pathogenicity of these unique class 3 VUS in the different genes involved in DSD. Functional validation of variants identified in novel genes may be more attractive in a research context. However, for individual families with VUS in well-established DSD genes such as AR or HSD17B3, functional analysis may provide a confirmed diagnosis that implies for relatives the option of undergoing their own DNA analysis and estimating the genetic risk of their own future offspring. This makes genetic follow-up important in these cases and demonstrates the usefulness of international databases and networks and the centralisation of functional studies of genetic variants in order to reduce costs and maximise expertise.The same is true for histopathological description, germ-cell tumour risk assessment in specific forms of DSD and classification of gonadal samples. Germ-cell tumour risk is related to the type of DSD (among other factors), but it is impossible to make risk estimates in individual cases.41–44 Gonadectomy may be indicated in cases with high-risk dysgenetic abdominal gonads that cannot be brought into a stable superficial (ie, inguinal, labioscrotal) position that allows clinical or radiological surveillance, or to avoid virilisation due to 5-alpha reductase deficiency in a 46,XY girl with a stable female gender identity.45 Pathological examination of DSD gonads requires specific expertise.

For example, the differentiation between benign germ cell abnormalities, such as delayed maturation and (pre)malignant development of germ cells, is crucial for clinical management but can be very troublesome.46 Centralised pathological examination of gonadal biopsy and gonadectomy samples in one centre, or a restricted number of centres, on a national scale can help to overcome the problem of non-uniform classification and has proven feasible in the Netherlands and Belgium. We therefore believe that uniform assessment and classification of gonadal differentiation patterns should also be addressed in guidelines on DSD management.International databases of gonadal tissues are crucial for learning more about the risk of malignancy in different forms of DSD, but they are only reliable if uniform criteria for histological classification are strictly applied.46 These criteria could be incorporated in many existing networks such as the I-DSD consortium, the Disorders of Sex Development Translational Research Network, the European Reference Network on Urogenital Diseases (eUROGEN), the EndoERN and COST actions.15–17 47Communication at the transition from paediatric to adult carePaediatric and adult teams need to collaborate closely to facilitate a well-organised transition from paediatric to adult specialist care.15 48–50 Both teams need to exchange information optimally and should consider transition as a longitudinal process rather than a fixed moment in time. Age-appropriate information is key at all ages, and an overview of topics to be discussed at each stage is described by Cools et al.15 Table 1 shows an example of how transition can be organised.View this table:Table 1 Example of transition table as used in the DSD clinic of the Erasmus Medical CenterPsychological support and the continued provision of information remains important for individuals with a DSD at all ages.15 22 In addition to the information given by the DSD team members, families and patients can benefit from resources such as support groups and information available on the internet.47 Information available online should be checked for accuracy and completeness when referring patients and parents to internet sites.Recommendations for future actionsMost guidelines and articles on the diagnosis and management of DSD are aimed at specialists and are only published in specialist journals or on websites for endocrinologists, urologists or geneticists. Yet there is a need for guidelines directed towards first-line and second-line healthcare workers that summarise the recommendations about the first crucial steps in the management of DSD. These should be published in widely available general medical journals and online, along with a national list of DSD centres.

Furthermore, DSD (expert) centres should provide continuous education to all those who may be involved in the identification of individuals with a DSD in order to enable these healthcare professionals to recognise atypical genitalia, to promptly refer individuals who have a DSD and to inform the patient and parents about this and subsequent diagnostic procedures.As DSD continues to be a rare condition, it will take time to evaluate the effects of having such a guideline on the preparedness of first-line and second-line healthcare workers to recognise DSD conditions. One way to evaluate this might be the development and use of questionnaires asking patients, carers and families and referring physicians how satisfied they were with the initial medical consultation and referral and what could be improved. A helpful addition to existing international databases that collect information on genetic variations would be a list of centres that offer suitable functional studies for certain genes, ideally covering the most frequently mutated genes (at minimum).Patient organisations can also play an important role in informing patients about newly available diagnostic or therapeutic strategies and options, and their influence and specific role has now been recognised and discussed in several publications.17 47 However, it should be kept in mind that these organisations do not represent all patients, as a substantial number of patients and parents are not member of such an organisation.Professionals have to provide optimal medical care based on well-established evidence, or at least on broad consensus. Yet not everything can be regulated by recommendations and guidelines. Options, ideas and wishes should be openly discussed between professionals, patients and families within their confidential relationship.

This will enable highly individualised holistic care tailored to the patient’s needs and expectations. Once they are well-informed of all available options, parents and/or patients can choose what they consider the optimal care for their children or themselves.15 16ConclusionThe Dutch-Flemish guideline uniquely addresses some topics that are under-represented in the literature, thus adding some key aspects to those addressed in recent consensus papers and guidelines.15–17 33 47As more children with a DSD are now being identified prenatally, and the literature on prenatal diagnosis of DSD remains scarce,20 21 we propose a prenatal diagnostic algorithm and emphasise the importance of having a prenatal specialist involved in or collaborating with DSD (expert) centres.We also stress that good communication between all involved parties is essential. Professionals should be well informed about protocols and communication. Collaboration between centres is necessary to optimise aspects of care such as uniform interpretation of gonadal pathology and functional testing of class 3 variants found by genetic testing. Guidelines can provide a framework within which individualised patient care should be discussed with all stakeholders.AcknowledgmentsThe authors would like to thank the colleagues of the DSD teams for their input in and critical reading of the Dutch-Flemish guideline.

Amsterdam University Center (AMC and VU), Maastricht University Medical Center, Erasmus Medical Center Rotterdam, Radboud University Medical Center Nijmegen, University Medical Center Groningen, University Medical Center Utrecht, Ghent University Hospital. The authors would like to thank Kate McIntyre for editing the revised manuscript and Tom de Vries Lentsch for providing the figures as a PDF. Three of the authors of this publication are members of the European Reference Network for rare endocrine diseases—Project ID 739543.IntroductionEndometrial cancer is the most common gynaecological malignancy in the developed world.1 Its incidence has risen over the last two decades as a consequence of the ageing population, fewer hysterectomies for benign disease and the obesity epidemic. In the USA, it is estimated that women have a 1 in 35 lifetime risk of endometrial cancer, and in contrast to cancers of most other sites, cancer-specific mortality has risen by approximately 2% every year since 2008 related to the rapidly rising incidence.2Endometrial cancer has traditionally been classified into type I and type II based on morphology.3 The more common subtype, type I, is mostly comprised of endometrioid tumours and is oestrogen-driven, arises from a hyperplastic endometrium, presents at an early stage and has an excellent 5 year survival rate.4 By contrast, type II includes non-endometrioid tumours, specifically serous, carcinosarcoma and clear cell subtypes, which are biologically aggressive tumours with a poor prognosis that are often diagnosed at an advanced stage.5 Recent efforts have focused on a molecular classification system for more accurate categorisation of endometrial tumours into four groups with distinct prognostic profiles.6 7The majority of endometrial cancers arise through the interplay of familial, genetic and lifestyle factors. Two inherited cancer predisposition syndromes, Lynch syndrome and the much rarer Cowden syndrome, substantially increase the lifetime risk of endometrial cancer, but these only account for around 3–5% of cases.8–10 Having first or second degree relative(s) with endometrial or colorectal cancer increases endometrial cancer risk, although a large European twin study failed to demonstrate a strong heritable link.11 The authors failed to show that there was greater concordance in monozygotic than dizygotic twins, but the study was based on relatively small numbers of endometrial cancers.

Lu and colleagues reported an association between common single nucleotide polymorphisms (SNPs) and endometrial cancer risk, revealing the potential role of SNPs in explaining part of the risk in both the familial and general populations.12 Thus far, many SNPs have been reported to modify susceptibility to endometrial cancer. However, much of this work predated genome wide association studies and is of variable quality. Understanding genetic predisposition to endometrial cancer could facilitate personalised risk assessment with a view to targeted prevention and screening interventions.13 This emerged as the most important unanswered research question in endometrial cancer according to patients, carers and healthcare professionals in our recently completed James Lind Womb Cancer Alliance Priority Setting Partnership.14 It would be particularly useful for non-endometrioid endometrial cancers, for which advancing age is so far the only predictor.15We therefore conducted a comprehensive systematic review of the literature to provide an overview of the relationship between SNPs and endometrial cancer risk. We compiled a list of the most robust endometrial cancer-associated SNPs. We assessed the applicability of this panel of SNPs with a theoretical polygenic risk score (PRS) calculation.

We also critically appraised the meta-analyses investigating the most frequently reported SNPs in MDM2. Finally, we described all SNPs reported within genes and pathways that are likely involved in endometrial carcinogenesis and metastasis.MethodsOur systematic review follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) collaboration 2009 recommendations. The registered protocol is available through PROSPERO (CRD42018091907).16Search strategyWe searched Embase, MEDLINE and Cumulative Index to Nursing and Allied Health Literature (CINAHL) databases via the Healthcare Databases Advanced Search (HDAS) platform, from 2007 to 2018, to identify studies reporting associations between polymorphisms and endometrial cancer risk. Key words including MeSH (Medical Subject Heading) terms and free-text words were searched in both titles and abstracts. The following terms were used.

€œendomet*”,“uter*”, “womb”, “cancer(s)”, “neoplasm(s)”, “endometrium tumour”, “carcinoma”, “adenosarcoma”, “clear cell carcinoma”, “carcinosarcoma”, “SNP”, “single nucleotide polymorphism”, “GWAS”, and “genome-wide association study/ies”. No other restrictions were applied. The search was repeated with time restrictions between 2018 and June 2019 to capture any recent publications.Eligibility criteriaStudies were selected for full-text evaluation if they were primary articles investigating a relationship between endometrial cancer and SNPs. Study outcome was either the increased or decreased risk of endometrial cancer relative to controls reported as an odds ratio (OR) with corresponding 95% confidence intervals (95% CIs).Study selectionThree independent reviewers screened all articles uploaded to a screening spreadsheet developed by Helena VonVille.17 Disagreements were resolved by discussion. Chronbach’s α score was calculated between reviewers and indicated high consistency at 0.92.

Case–control, prospective and retrospective studies, genome-wide association studies (GWAS), and both discovery and validation studies were selected for full-text evaluation. Non-English articles, editorials, conference abstracts and proceedings, letters and correspondence, case reports and review articles were excluded.Candidate-gene studies with at least 100 women and GWAS with at least 1000 women in the case arm were selected to ensure reliability of the results, as explained by Spencer et al.18 To construct a panel of up to 30 SNPs with the strongest evidence of association, those with the strongest p values were selected. For the purpose of an SNP panel, articles utilising broad European or multi-ethnic cohorts were selected. Where overlapping populations were identified, the most comprehensive study was included.Data extraction and synthesisFor each study, the following data were extracted. SNP ID, nearby gene(s)/chromosome location, OR (95% CI), p value, minor or effect allele frequency (MAF/EAF), EA (effect allele) and OA (other allele), adjustment, ethnicity and ancestry, number of cases and controls, endometrial cancer type, and study type including discovery or validation study and meta-analysis.

For risk estimates, a preference towards most adjusted results was applied. For candidate-gene studies, a standard p value of<0.05 was applied and for GWAS a p value of <5×10-8, indicating genome-wide significance, was accepted as statistically significant. However, due to the limited number of SNPs with p values reaching genome-wide significance, this threshold was then lowered to <1×10-5, allowing for marginally significant SNPs to be included. As shown by Mavaddat et al, for breast cancer, SNPs that fall below genome-wide significance may still be useful for generating a PRS and improving the models.19We estimated the potential value of a PRS based on the most significant SNPs by comparing the predicted risk for a woman with a risk score in the top 1% of the distribution to the mean predicted risk. Per-allele ORs and MAFs were taken from the publications and standard errors (SEs) for the lnORs were derived from published 95% CIs.

The PRS was assumed to have a Normal distribution, with mean 2∑βipI and SE, σ, equal to √2∑βi2pI(1−pi), according to the binomial distribution, where the summation is over all SNPs in the risk score. Hence the relative risk (RR) comparing the top 1% of the distribution to the mean is given by exp(Z0.01σ), where Z is the inverse of the standard normal cumulative distribution.ResultsThe flow chart of study selection is illustrated in figure 1. In total, 453 text articles were evaluated and, of those, 149 articles met our inclusion criteria. One study was excluded from table 1, for having an Asian-only population, as this would make it harder to compare with the rest of the results which were all either multi-ethnic or Caucasian cohorts, as stated in our inclusion criteria for the SNP panel.20 Any SNPs without 95% CIs were also excluded from any downstream analysis. Additionally, SNPs in linkage disequilibrium (r2 >0.2) with each other were examined, and of those in linkage disequilibrium, the SNP with strongest association was reported.

Per allele ORs were used unless stated otherwise.View this table:Table 1 List of top SNPs most likely to contribute to endometrial cancer risk identified through systematic review of recent literature21–25Study selection flow diagram. *Reasons. Irrelevant articles, articles focusing on other conditions, non-GWAS/candidate-gene study related articles, technical and duplicate articles. GWAS, genome-wide association study. Adapted from.

Moher D, Liberati A, Tetzlaff J, Altman DG, The PRISMA Group (2009). Preferred Reporting Items for Systematic Reviews and Meta-Analyses. The PRISMA Statement. PLoS Med 6(6). E1000097.

Doi:10.1371/journal.pmed1000097." data-icon-position data-hide-link-title="0">Figure 1 Study selection flow diagram. *Reasons. Irrelevant articles, articles focusing on other conditions, non-GWAS/candidate-gene study related articles, technical and duplicate articles. GWAS, genome-wide association study. Adapted from.

Moher D, Liberati A, Tetzlaff J, Altman DG, The PRISMA Group (2009). Preferred Reporting Items for Systematic Reviews and Meta-Analyses. The PRISMA Statement. PLoS Med 6(6). E1000097.

Doi:10.1371/journal.pmed1000097.Top SNPs associated with endometrial cancer riskFollowing careful interpretation of the data, 24 independent SNPs with the lowest p values that showed the strongest association with endometrial cancer were obtained (table 1).21–25 These SNPs are located in or around genes coding for transcription factors, cell growth and apoptosis regulators, and enzymes involved in the steroidogenesis pathway. All the SNPs presented here were reported on the basis of a GWAS or in one case, an exome-wide association study, and hence no SNPs from candidate-gene studies made it to the list. This is partly due to the nature of larger GWAS providing more comprehensive and powered results as opposed to candidate gene studies. Additionally, a vast majority of SNPs reported by candidate-gene studies were later refuted by large-scale GWAS such as in the case of TERT and MDM2 variants.26 27 The exception to this is the CYP19 gene, where candidate-gene studies reported an association between variants in this gene with endometrial cancer in both Asian and broad European populations, and this association was more recently confirmed by large-scale GWAS.21 28–30 Moreover, a recent article authored by O’Mara and colleagues reviewed the GWAS that identified most of the currently known SNPs associated with endometrial cancer.31Most of the studies represented in table 1 are GWAS and the majority of these involved broad European populations. Those having a multi-ethnic cohort also consisted primarily of broad European populations.

Only four of the variants in table 1 are located in coding regions of a gene, or in regulatory flanking regions around the gene. Thus, most of these variants would not be expected to cause any functional effects on the gene or the resulting protein. An eQTL search using GTEx Portal showed that some of the SNPs are significantly associated (p<0.05) with modified transcription levels of the respective genes in various tissues such as prostate (rs11263761), thyroid (rs9668337), pituitary (rs2747716), breast mammary (rs882380) and testicular (rs2498794) tissue, as summarised in table 2.View this table:Table 2 List of eQTL hits for the selected panel of SNPsThe only variant for which there was an indication of a specific association with non-endometrioid endometrial cancer was rs148261157 near the BCL11A gene. The A allele of this SNP had a moderately higher association in the non-endometrioid arm (OR 1.64, 95% CI 1.32 to 2.04. P=9.6×10-6) compared with the endometrioid arm (OR 1.25, 95% CI 1.14 to 1.38.

P=4.7×10-6).21Oestrogen receptors α and β encoded by ESR1 and ESR2, respectively, have been extensively studied due to the assumed role of oestrogens in the development of endometrial cancer. O’Mara et al reported a lead SNP (rs79575945) in the ESR1 region that was associated with endometrial cancer (p=1.86×10-5).24 However, this SNP did not reach genome-wide significance in a more recent larger GWAS.21 No statistically significant associations have been reported between endometrial cancer and SNPs in the ESR2 gene region.AKT is an oncogene linked to endometrial carcinogenesis. It is involved in the PI3K/AKT/mTOR pro-proliferative signalling pathway to inactivate apoptosis and allow cell survival. The A allele of rs2494737 and G allele of rs2498796 were reported to be associated with increased and decreased risk of endometrial cancer in 2016, respectively.22 30 However, this association was not replicated in a larger GWAS in 2018.21 Nevertheless, given the previous strong indications, and biological basis that could explain endometrial carcinogenesis, we decided to include an AKT1 variant (rs2498794) in our results.PTEN is a multi-functional tumour suppressor gene that regulates the AKT/PKB signalling pathway and is commonly mutated in many cancers including endometrial cancer.32 Loss-of-function germline mutations in PTEN are responsible for Cowden syndrome, which exerts a lifetime risk of endometrial cancer of up to 28%.9 Lacey and colleagues studied SNPs in the PTEN gene region. However, none showed significant differences in frequency between 447 endometrial cancer cases and 439 controls of European ancestry.33KRAS mutations are known to be present in endometrial cancer.

These can be activated by high levels of KLF5 (transcriptional activator). Three SNPs have been identified in or around KLF5 that are associated with endometrial cancer. The G allele of rs11841589 (OR 1.15, 95% CI 1.11 to 1.21. P=4.83×10-11), the A allele of rs9600103 (OR 1.23, 95% CI 1.16 to 1.30. P=3.76×10-12) and C allele of rs7981863 (OR 1.16, 95% CI 1.12 to 1.20.

P=2.70×10-17) have all been found to be associated with an increased likelihood of endometrial cancer in large European cohorts.21 30 34 It is worth noting that these SNPs are not independent, and hence they quite possibly tag the same causal variant.The MYC family of proto-oncogenes encode transcription factors that regulate cell proliferation, which can contribute to cancer development if dysregulated. The recent GWAS by O’Mara et al reported three SNPs within the MYC region that reached genome-wide significance with conditional p values reaching at least 5×10–8.35To test the utility of these SNPs as predictive markers, we devised a theoretical PRS calculation using the log ORs and EAFs per SNP from the published data. The results were very encouraging with an RR of 3.16 for the top 1% versus the mean, using all the top SNPs presented in table 1 and 2.09 when using only the SNPs that reached genome-wide significance (including AKT1).Controversy surrounding MDM2 variant SNP309MDM2 negatively regulates tumour suppressor gene TP53, and as such, has been extensively studied in relation to its potential role in predisposition to endometrial cancer. Our search identified six original studies of the association between MDM2 SNP rs2279744 (also referred to as SNP309) and endometrial cancer, all of which found a statistically significant increased risk per copy of the G allele. Two more original studies were identified through our full-text evaluation.

However, these were not included here as they did not meet our inclusion criteria—one due to small sample size, the other due to studying rs2279744 status dependent on another SNP.36 37 Even so, the two studies were described in multiple meta-analyses that are listed in table 3. Different permutations of these eight original studies appear in at least eight published meta-analyses. However, even the largest meta-analysis contained <2000 cases (table 3)38View this table:Table 3 Characteristics of studies that examined MDM2 SNP rs2279744In comparison, a GWAS including nearly 13 000 cases found no evidence of an association with OR and corresponding 95% CI of 1.00 (0.97 to 1.03) and a p value of 0.93 (personal communication).21 Nevertheless, we cannot completely rule out a role for MDM2 variants in endometrial cancer predisposition as the candidate-gene studies reported larger effects in Asians, whereas the GWAS primarily contained participants of European ancestry. There is also some suggestion that the SNP309 variant is in linkage disequilibrium with another variant, SNP285, which confers an opposite effect.It is worth noting that the SNP285C/SNP309G haplotype frequency was observed in up to 8% of Europeans, thus requiring correction for the confounding effect of SNP285C in European studies.39 However, aside from one study conducted by Knappskog et al, no other study including the meta-analyses corrected for the confounding effect of SNP285.40 Among the studies presented in table 3, Knappskog et al (2012) reported that after correcting for SNP285, the OR for association of this haplotype with endometrial cancer was much lower, though still significant. Unfortunately, the meta-analyses which synthesised Knappskog et al (2012), as part of their analysis, did not correct for SNP285C in the European-based studies they included.38 41 42 It is also concerning that two meta-analyses using the same primary articles failed to report the same result, in two instances.38 42–44DiscussionThis article represents the most comprehensive systematic review to date, regarding critical appraisal of the available evidence of common low-penetrance variants implicated in predisposition to endometrial cancer.

We have identified the most robust SNPs in the context of endometrial cancer risk. Of those, only 19 were significant at genome-wide level and a further five were considered marginally significant. The largest GWAS conducted in this field was the discovery- and meta-GWAS by O’Mara et al, which utilised 12 096 cases and 108 979 controls.21 Despite the inclusion of all published GWAS and around 5000 newly genotyped cases, the total number did not reach anywhere near what is currently available for other common cancers such as breast cancer. For instance, BCAC (Breast Cancer Association Consortium) stands at well over 200 000 individuals with more than half being cases, and resulted in identification of ~170 SNPs in relation to breast cancer.19 45 A total of 313 SNPs including imputations were then used to derive a PRS for breast cancer.19 Therefore, further efforts should be directed to recruit more patients, with deep phenotypic clinical data to allow for relevant adjustments and subgroup analyses to be conducted for better precision.A recent pre-print study by Zhang and colleagues examined the polygenicity and potential for SNP-based risk prediction for 14 common cancers, including endometrial cancer, using available summary-level data from European-ancestry datasets.46 They estimated that there are just over 1000 independent endometrial cancer susceptibility SNPs, and that a PRS comprising all such SNPs would have an area under the receiver-operator curve of 0.64, similar to that predicted for ovarian cancer, but lower than that for the other cancers in the study. The modelling in the paper suggests that an endometrial cancer GWAS double the size of the current largest study would be able to identify susceptibility SNPs together explaining 40% of the genetic variance, but that in order to explain 75% of the genetic variance it would be necessary to have a GWAS comprising close to 150 000 cases and controls, far in excess of what is currently feasible.We found that the literature consists mainly of candidate-gene studies with small sample sizes, meta-analyses reporting conflicting results despite using the same set of primary articles, and multiple reports of significant SNPs that have not been validated by any larger GWAS.

The candidate-gene studies were indeed the most useful and cheaper technique available until the mid to late 2000s. However, a lack of reproducibility (particularly due to population stratification and reporting bias), uncertainty of reported associations, and considerably high false discovery rates make these studies much less appropriate in the post-GWAS era. Unlike the candidate-gene approach, GWAS do not require prior knowledge, selection of genes or SNPs, and provide vast amounts of data. Furthermore, both the genotyping process and data analysis phases have become cheaper, the latter particularly due to faster and open-access pre-phasing and imputation tools being made available.It is clear from table 1 that some SNPs were reported with wide 95% CI, which can be directly attributed to small sample sizes particularly when restricting the cases to non-endometrioid histology only, low EAF or poor imputation quality. Thus, these should be interpreted with caution.

Additionally, most of the SNPs reported by candidate-gene studies were not detected by the largest GWAS to date conducted by O’Mara et al.21 However, this does not necessarily mean that the possibility of those SNPs being relevant should be completely dismissed. Moreover, meta-analyses were attempted for other variants. However, these showed no statistically significant association and many presented with high heterogeneity between the respective studies (data not shown). Furthermore, as many studies utilised the same set of cases and/or controls, conducting a meta-analysis was not possible for a good number of SNPs. It is therefore unequivocal that the literature is crowded with numerous small candidate-gene studies and conflicting data.

This makes it particularly hard to detect novel SNPs and conduct meaningful meta-analyses.We found convincing evidence for 19 variants that indicated the strongest association with endometrial cancer, as shown in table 1. The associations between endometrial cancer and variants in or around HNF1B, CYP19A1, SOX4, MYC, KLF and EIF2AK found in earlier GWAS were then replicated in the latest and largest GWAS. These SNPs showed promising potential in a theoretical PRS we devised based on published data. Using all 24 or genome-wide significant SNPs only, women with a PRS in the top 1% of the distribution would be predicted to have a risk of endometrial cancer 3.16 and 2.09 times higher than the mean risk, respectively.However, the importance of these variants and relevance of the proximate genes in a functional or biological context is challenging to evaluate. Long distance promoter regulation by enhancers may disguise the genuine target gene.

In addition, enhancers often do not loop to the nearest gene, further complicating the relevance of nearby gene(s) to a GWAS hit. In order to elucidate biologically relevant candidate target genes in endometrial cancer, O’Mara et al looked into promoter-associated chromatin looping using a modern HiChIP approach.47 The authors utilised normal and tumoural endometrial cell lines for this analysis which showed significant enrichment for endometrial cancer heritability, with 103 candidate target genes identified across the 13 risk loci identified by the largest ECAC GWAS. Notable genes identified here were CDKN2A and WT1, and their antisense counterparts. The former was reported to be nearby of rs1679014 and the latter of rs10835920, as shown in table 1. Moreover, of the 36 candidate target genes, 17 were found to be downregulated while 19 were upregulated in endometrial tumours.The authors also investigated overlap between the 13 endometrial cancer risk loci and top eQTL variants for each target gene.47 In whole blood, of the two particular lead SNPs, rs8822380 at 17q21.32 was a top eQTL for SNX11 and HOXB2, whereas rs937213 at 15q15.1 was a top eQTL for SRP14.

In endometrial tumour, rs7579014 at 2p16.1 was found to be a top eQTL for BCL11A. This is particularly interesting because BCL11A was the only nearby/candidate gene that had a GWAS association reported in both endometrioid and non-endometrioid subtypes. The study looked at protein–protein interactions between endometrial cancer drivers and candidate target gene products. Significant interactions were observed with TP53 (most significant), AKT, PTEN, ESR1 and KRAS, among others. Finally, when 103 target candidate genes and 387 proteins were combined together, 462 pathways were found to be significantly enriched.

Many of these are related to gene regulation, cancer, obesity, insulinaemia and oestrogen exposure. This study clearly showed a potential biological relevance for some of the SNPs reported by ECAC GWAS in 2018.Most of the larger included studies used cohorts primarily composed of women of broad European descent. Hence, there are negligible data available for other ethnicities, particularly African women. This is compounded by the lack of reference genotype data available for comparative analysis, making it harder for research to be conducted in ethnicities other than Europeans. This poses a problem for developing risk prediction models that are equally valuable and predictive across populations.

Thus, our results also are of limited applicability to non-European populations.Furthermore, considering that non-endometrioid cases comprise a small proportion (~20%) of all endometrial cancer cases, much larger cohort sizes are needed to detect any genuine signals for non-endometrioid tumours. Most of the evaluated studies looked at either overall/mixed endometrial cancer subtypes or endometrioid histology, and those that looked at variant associations with non-endometrioid histology were unlikely to have enough power to detect any signal with statistical significance. This is particularly concerning because non-endometrioid subtypes are biologically aggressive tumours with a much poorer prognosis that contribute disproportionately to mortality from endometrial cancer. It is particularly important that attempts to improve early detection and prevention of endometrial cancer focus primarily on improving outcomes from these subtypes. It is also worth noting that, despite the current shift towards a molecular classification of endometrial cancer, most studies used the overarching classical Bokhman’s classification system, type I versus type II, or no histological classification system at all.

Therefore, it is important to create and follow a standardised and comprehensive classification system for reporting tumour subtypes for future studies.This study compiled and presented available information for an extensively studied, yet unproven in large datasets, SNP309 variant in MDM2. Currently, there is no convincing evidence for an association between this variant and endometrial cancer risk. Additionally, of all the studies, only one accounted for the opposing effect of a nearby variant SNP285 in their analyses. Thus, we conclude that until confirmed by a sufficiently large GWAS, this variant should not be considered significant in influencing the risk of endometrial cancer and therefore not included in a PRS. This is also true for the majority of the SNPs reported in candidate-gene studies, as the numbers fall far short of being able to detect genuine signals.This systematic review presents the most up-to-date evidence for endometrial cancer susceptibility variants, emphasising the need for further large-scale studies to identify more variants of importance, and validation of these associations.

Until data from larger and more diverse cohorts are available, the top 24 SNPs presented here are the most robust common genetic variants that affect endometrial cancer risk. The multiplicative effects of these SNPs could be used in a PRS to allow personalised risk prediction models to be developed for targeted screening and prevention interventions for women at greatest risk of endometrial cancer..

IntroductionIn recent years, many studies have been published on new diagnostic possibilities and management approaches in cohorts of patients suspected to have a disorder/difference of sex development low cost vibramycin (DSD).1–13 Based on these studies, it has become clear that services and institutions still differ in the composition of the multidisciplinary teams that provide care for patients who have a DSD.11 14 Several projects have now worked to resolve this variability in care. The European Cooperation in Science and Technology (EU COST) action BM1303 ‘A systematic elucidation of differences of sex development’ has been a low cost vibramycin platform to achieve European agreement on harmonisation of clinical management and laboratory practices.15–17 Another such initiative involved an update of the 2006 DSD consensus document by an international group of professionals and patient representatives.18 These initiatives have highlighted how cultural and financial aspects and the availability of resources differ significantly between countries and societies, a situation that hampers supranational agreement on common diagnostic protocols. As only a few national guidelines have been published in international journals, comparison of these guidelines is difficult even though such a comparison is necessary to capture the differences and initiate actions to overcome them. Nonetheless, four DSD (expert) centres located in the Netherlands and Flanders (the Dutch-speaking Northern part of Belgium) have collaborated to produce a detailed guideline on diagnostics in DSD.19 This shows that a supranational guideline can be a reasonable approach for low cost vibramycin countries with similarly structured healthcare systems and similar resources.

Within the guideline there is agreement that optimisation of expertise and care can be achieved through centralisation, for example, by limiting analysis of next-generation sequencing (NGS)-based diagnostic panels to only a few centres and by centralising pathological review of gonadal tissues. International networks such as the low cost vibramycin European Reference Network for rare endocrine conditions (EndoERN), in which DSD is embedded, may facilitate the expansion of this kind of collaboration across Europe.This paper highlights key discussion points in the Dutch-Flemish guideline that have been insufficiently addressed in the literature thus far because they reflect evolving technologies or less visible stakeholders. For example, prenatal observation of an atypical aspect of the genitalia indicating a possible DSD is becoming increasingly common, and we discuss appropriate counselling and a diagnostic approach for these cases, including the option of using NGS-based genetic testing. So far, little attention has been paid to this process.20 21 Furthermore, informing patients and/or their parents about atypical sex development and why this may warrant referral to a specialised team may be challenging, especially for professionals low cost vibramycin with limited experience in DSD.22 23 Therefore, a section of the Dutch-Flemish guideline was written for these healthcare providers.

Moreover, this enables DSD specialists to refer to the guideline when advising a referral. Transition from the prenatal low cost vibramycin to the postnatal team and from the paediatric to the adult team requires optimal communication between the specialists involved. Application of NGS-based techniques may lead to a higher diagnostic yield, providing a molecular genetic diagnosis in previously unsolved cases.16 We address the timing of this testing and the problems associated with this technique such as the interpretation of variants of unknown clinical significance (VUS). Similarly, histopathological interpretation and classification of removed gonadal tissue is challenging and would benefit from low cost vibramycin international collaboration and centralisation of expertise.MethodsFor the guideline revision, an interdisciplinary multicentre group was formed with all members responsible for updating the literature for a specific part of the guideline.

Literature search in PubMed was not systematic, but rather intended to be broad in order to cover all areas and follow expert opinions. This approach is more in line with the Clinical Practice Advisory Document method described by Burke et al24 for guidelines involving low cost vibramycin genetic practice because it is often troublesome to substantiate such guidelines with sufficient evidence due to the rapid changes in testing methods, for example, gene panels. All input provided by the group was synthesised by the chairperson (YvB), who also reviewed abstracts of papers on DSD published between 2010 and September 2017 for the guideline and up to October 2019 for this paper. Abstracts had to be written in English and were identified using a broad range of Medical Subject Headings terms (eg, DSD, genetic, review, diagnosis, low cost vibramycin diagnostics, 46,XX DSD, 46,XY DSD, guideline, multidisciplinary care).

Next, potentially relevant papers on diagnostic procedures in DSD were selected. Case reports were excluded, as were articles that were not open access or retrievable through low cost vibramycin institutional access. Based on this, a draft guideline was produced that was in line with the international principles of good diagnostic care in DSD. This draft was discussed by the writing committee and, after having obtained agreement on remaining points of discussion, revised into a low cost vibramycin final draft.

This version was sent to a broad group of professionals from academic centres and DSD teams whose members had volunteered to review the draft guideline. After receiving and incorporating their input, the final version was presented low cost vibramycin to the paediatric and genetic associations for approval. After approval by the members of the paediatric (NVK), clinical genetic (VKGN) and genetic laboratory (VKGL) associations, the guideline was published on their respective websites.19 Although Turner syndrome and Klinefelter syndrome are considered to be part of the DSD spectrum, they are not extensively discussed in this diagnostic guideline as guidelines dedicated to these syndromes already exist.25 26 However, some individuals with Turner syndrome or Klinefelter syndrome may present with ambiguous or atypical genitalia and may therefore initially follow the DSD diagnostic process.Guideline highlightsPrenatal settingPresentationThe most frequent prenatal presentation of a DSD condition is atypical genitalia found on prenatal ultrasound as an isolated finding or in combination with other structural anomalies. This usually occurs low cost vibramycin after the 20-week routine medical ultrasound for screening of congenital anomalies, but may also occur earlier, for example, when a commercial ultrasound is performed at the request of the parents.Another way DSD can be diagnosed before birth is when invasive prenatal genetic testing carried out for a different reason, for example, due to suspicion of other structural anomalies, reveals a discrepancy between the genotypic sex and the phenotypic sex seen by ultrasound.

In certified laboratories, the possibility of a sample switch is extremely low but should be ruled out immediately. More often, the discrepancy low cost vibramycin will be due to sex-chromosome mosaicism or a true form of DSD.A situation now occurring with increasing frequency is a discrepancy between the genotypic sex revealed by non-invasive prenatal testing (NIPT), which is now available to high-risk pregnant women in the Netherlands and to all pregnant women in Belgium, and later ultrasound findings. NIPT screens for CNVs in the fetus. However, depending on legal restrictions and/or ethical considerations, the X and low cost vibramycin Y chromosomes are not always included in NIPT analysis and reports.

If the X and Y chromosomes are included, it is important to realise that the presence of a Y-chromosome does not necessarily imply male fetal development. At the time that NIPT is performed (usually 11–13 weeks), genital development cannot be reliably appreciated by ultrasound, so any discrepancy or atypical aspect of the genitalia will only be noticed later in pregnancy and low cost vibramycin should prompt further evaluation.Counselling and diagnosticsIf a DSD is suspected, first-line sonographers and obstetricians should refer the couple to their colleague prenatal specialists working with or in a DSD team. After confirming an atypical genital on ultrasound, the specialist team should offer the couple a referral for genetic counselling to discuss the possibility of performing invasive prenatal testing (usually an amniocentesis) to identify an underlying cause that fits the ultrasound findings.22 23 To enable the parents to make a well-informed decision, prenatal counselling should, in our opinion, include. Information on the ultrasound findings and low cost vibramycin the limitations of this technique.

The procedure(s) that can be followed, including the risks associated with an amniocentesis. And the type of information low cost vibramycin genetic testing can and cannot provide. Knowing which information has been provided and what words have been used by the prenatal specialist is very helpful for those involved in postnatal care.It is important that parents understand that the biological sex of a baby is determined by a complex interplay of chromosomes, genes and hormones, and thus that assessment of the presence or absence of a Y-chromosome alone is insufficient to assign the sex of their unborn child or, as in any unborn child, say anything about the child’s future gender identity.Expecting parents can be counselled by the clinical geneticist and the psychologist from the DSD team, although other DSD specialists can also be involved. The clinical low cost vibramycin geneticist should be experienced in prenatal counselling and well informed about the diagnostic possibilities given the limited time span in which test results need to be available to allow parents to make a well-informed decision about whether or not to continue the pregnancy.

Termination of pregnancy can be considered, for instance, in a syndromic form of DSD low cost vibramycin with multiple malformations, but when the DSD occurs as an apparently isolated condition, expecting parents may also consider termination of pregnancy, which, although considered controversial by some, is legal in Belgium and the Netherlands. The psychologist of the DSD team can support parents during and after pregnancy and help them cope with feelings of uncertainty and eventual considerations of a termination of pregnancy, as well as with practical issues, for example, how to inform others. The stress of not low cost vibramycin knowing exactly what the child’s genitalia will look like and uncertainty about the diagnosis, treatment and prognosis cannot be avoided completely. Parents are informed that if the postnatal phenotype is different from what was prenatally expected, the advice given about diagnostic testing can be adjusted accordingly, for example, if a hypospadias is milder than was expected based on prenatal ultrasound images.

In our experience, parents appreciate having already spoken to some members of the DSD team during pregnancy and having low cost vibramycin a contact person before birth.After expert prenatal counselling, a significant number of pregnant couples decline prenatal testing (personal experience IALG, MK, ABD, YvB, MC and HC-vdG). At birth, umbilical cord blood is a good source for (molecular) karyotyping and storage of DNA and can be obtained by the obstetrician, midwife or neonatologist. The terminology used in communication with parents should be carefully chosen,22 23 and midwives and staff of neonatal and delivery units should be clearly instructed to use gender-neutral and non-stigmatising vocabulary (eg, ‘your baby’) as long as sex assignment is pending.An algorithm low cost vibramycin for diagnostic evaluation of a suspected DSD in the prenatal situation is proposed in figure 1. When couples opt for invasive prenatal diagnosis, the genetic analysis usually involves an (SNP)-array.

It was recently estimated that >30% of individuals who have a DSD have additional structural anomalies, with cardiac and neurological anomalies and fetal growth restriction being particularly common.27 28 If additional anomalies are seen, the geneticist can consider low cost vibramycin specific gene defects that may underlie a known genetic syndrome or carry out NGS. NGS-based techniques have also now made their appearance in prenatal diagnosis of congenital anomalies.29 30 Panels using these techniques can be specific for genes involved in DSD, or be larger panels covering multiple congenital anomalies, and are usually employed with trio-analysis to compare variants identified in the child with the parents’ genetics.29–31 Finding a genetic cause before delivery can help reduce parental stress in the neonatal period and speed up decisions regarding gender assignment. In such cases there is no tight time limit, and we propose completing the analysis well before the expected delivery.Disorders/differences of sex development (DSD) in the prenatal low cost vibramycin setting. A diagnostic algorithm.

*SOX9. Upstream anomalies and balanced translocations at promotor sites!. Conventional karyotyping can be useful. NGS, next-generation sequencing." data-icon-position data-hide-link-title="0">Figure 1 Disorders/differences of sex development (DSD) in the prenatal setting.

A diagnostic algorithm. *SOX9. Upstream anomalies and balanced translocations at promotor sites!. Conventional karyotyping can be useful.

NGS, next-generation sequencing.First contact by a professional less experienced in DSDWhereas most current guidelines start from the point when an individual has been referred to the DSD team,1 15 the Dutch-Flemish guideline dedicates a chapter to healthcare professionals less experienced in DSD as they are often the first to suspect or identify such a condition. Apart from the paper of Indyk,7 little guidance is available for these professionals about how to act in such a situation. The chapter in the Dutch-Flemish guideline summarises the various clinical presentations that a DSD can have and provides information on how to communicate with parents and/or patients about the findings of the physical examination, the first-line investigations and the need for prompt referral to a specialised centre for further evaluation. Clinical examples are offered to illustrate some of these recurring situations.

The medical issues in DSD can be very challenging, and the social and psychological impact is high. For neonates with ambiguous genitalia, sex assignment is an urgent and crucial issue, and it is mandatory that parents are informed that it is possible to postpone registration of their child’s sex. In cases where sex assignment has already taken place, the message that the development of the gonads or genitalia is still atypical is complicated and distressing for patients and parents or carers. A list of contact details for DSD centres and patient organisations in the Netherlands and Flanders is attached to the Dutch-Flemish guideline.

Publishing such a list, either in guidelines or online, can help healthcare professionals find the nearest centres for consultations and provide patients and patient organisations with an overview of the centres where expertise is available.Timing and place of genetic testing using NGS-based gene panelsThe diagnostic workup that is proposed for 46,XX and 46,XY DSD is shown in figures 2 and 3, respectively. Even with the rapidly expanding molecular possibilities, a (family) history and a physical examination remain the essential first steps in the diagnostic process. Biochemical and hormonal screening aim at investigating serum electrolytes, renal function and the hypothalamic-pituitary-gonadal and hypothalamic-pituitary-adrenal axes. Ultrasound screening of kidneys and internal genitalia, as well as establishing genotypic sex, should be accomplished within 48 hours and complete the baseline diagnostic work-up of a child born with ambiguous genitalia.1 16 32 3346,XX disorders/differences of sex development (DSD) in the postnatal setting.

A diagnostic algorithm. NGS, next-generation sequencing. CAH, Congenital adrenal hyperplasia. AMH, Anti-Müllerian Hormone." data-icon-position data-hide-link-title="0">Figure 2 46,XX disorders/differences of sex development (DSD) in the postnatal setting.

A diagnostic algorithm. NGS, next-generation sequencing. CAH, Congenital adrenal hyperplasia. AMH, Anti-Müllerian Hormone.46,XY disorders/differences of sex development (DSD) in the postnatal setting.

A diagnostic algorithm. * SOX9. Upstream anomalies and balanced translocations at promotor sites!. Conventional karyotyping can be useful.

NGS, next-generation sequencing." data-icon-position data-hide-link-title="0">Figure 3 46,XY disorders/differences of sex development (DSD) in the postnatal setting. A diagnostic algorithm. *SOX9. Upstream anomalies and balanced translocations at promotor sites!.

Conventional karyotyping can be useful. NGS, next-generation sequencing.Very recently, a European position paper has been published focusing on the genetic workup of DSD.16 It highlights the limitations and drawbacks of NGS-based tests, which include the chance of missing subtle structural variants such as CNVs and mosaicism and the fact that NGS cannot detect methylation defects or other epigenetic changes.16 28 31 Targeted DNA analysis is preferred in cases where hormonal investigations suggest a block in steroidogenesis (eg, 11-β-hydroxylase deficiency, 21-hydroxylase deficiency), or in the context of a specific clinical constellation such as the often coincidental finding of Müllerian structures in a boy with normal external genitalia or cryptorchidism, that is, persistent Müllerian duct syndrome.33 34 Alternative tests should also be considered depending on the available information. Sometimes, a simple mouth swab for FISH analysis can detect mosaic XY/X in a male with hypospadias or asymmetric gonadal development or in a female with little or no Turner syndrome stigmata and a normal male molecular karyotyping profile or peripheral blood karyotype. Such targeted testing avoids incidental findings and is cheaper and faster than analysis of a large NGS-based panel, although the cost difference is rapidly declining.However, due to the genetic and phenotypic heterogeneity of DSD conditions, the most cost-effective next steps in the majority of cases are whole exome sequencing followed by panel analysis of genes involved in genital development and function or trio-analysis of a large gene panel (such as a Mendeliome).16 35–38 Pretest genetic counselling involves discussing what kind of information will be reported to patients or parents and the chance of detecting VUS, and the small risk of incidental findings when analysing a DSD panel should be mentioned.

Laboratories also differ in what class of variants they report.39 In our experience, the fear of incidental findings is a major reason why some parents refrain from genetic testing.Timing of the DSD gene panel analysis is also important. While some patients or parents prefer that all diagnostic procedures be performed as soon as possible, others need time to reflect on the complex information related to more extensive genetic testing and on its possible consequences. If parents or patients do not consent to panel-based genetic testing, analysis of specific genes, such as WT1, should be considered when appropriate in view of the clinical consequences if a mutation is present (eg, clinical surveillance of renal function and screening for Wilms’ tumour in the case of WT1 mutations). Genes that are more frequently involved in DSD (eg, SRY, NR5A1) and that match the specific clinical and hormonal features in a given patient could also be considered for sequencing.

Targeted gene analysis may also be preferred in centres located in countries that do not have the resources or technical requirements to perform NGS panel-based genetic testing. Alternatively, participation by these centres in international collaborative networks may allow them to outsource the molecular genetic workup abroad.Gene panels differ between centres and are regularly updated based on scientific progress. A comparison of DSD gene panels used in recent studies can be found at https://www.nature.com/articles/s41574-018-0010-8%23Sec46.15 The panels currently used at the coauthors’ institutions can be found on their respective websites. Given the pace of change, it is important to regularly consider repeating analysis in patients with an unexplained DSD, for example, when they transition into adult care or when they move from one centre to another.

This also applies to patients in whom a clinical diagnosis has never been genetically confirmed. Confusion may arise when the diagnosis cannot be confirmed or when a mutation is identified in a different gene, for example, NR5A1 in someone with a clinical diagnosis of CAIS that has other consequences for relatives. Hence, new genetic counselling should always accompany new diagnostic endeavours.Class 3 variants and histopathological examinationsThe rapidly evolving diagnostic possibilities raise new questions. What do laboratories report?.

How should we deal with the frequent findings of mainly unique VUS or class 3 variants (ACMG recommendation) in the many different DSD-related genes in the diagnostic setting?. Reporting VUS can be a source of uncertainty for parents, but not reporting these variants precludes further investigations to determine their possible pathogenicity. It can also be difficult to prove variant pathogenicity, both on gene-level and variant-level.39 Moreover, given the gonad-specific expression of some genes and the variable phenotypic spectrum and reduced penetrance, segregation analysis is not always informative. A class 3 variant that does not fit the clinical presentation may be unrelated to the observed phenotype, but it could also represent a newly emerging phenotype.

This was recently demonstrated by the identification of the NR5A1 mutation, R92W, in individuals with 46,XX testicular and ovotesticular DSD.40 This gene had previously been associated with 46,XY DSD. In diagnostic laboratories, there is usually no capacity or expertise to conduct large-scale functional studies to determine pathogenicity of these unique class 3 VUS in the different genes involved in DSD. Functional validation of variants identified in novel genes may be more attractive in a research context. However, for individual families with VUS in well-established DSD genes such as AR or HSD17B3, functional analysis may provide a confirmed diagnosis that implies for relatives the option of undergoing their own DNA analysis and estimating the genetic risk of their own future offspring.

This makes genetic follow-up important in these cases and demonstrates the usefulness of international databases and networks and the centralisation of functional studies of genetic variants in order to reduce costs and maximise expertise.The same is true for histopathological description, germ-cell tumour risk assessment in specific forms of DSD and classification of gonadal samples. Germ-cell tumour risk is related to the type of DSD (among other factors), but it is impossible to make risk estimates in individual cases.41–44 Gonadectomy may be indicated in cases with high-risk dysgenetic abdominal gonads that cannot be brought into a stable superficial (ie, inguinal, labioscrotal) position that allows clinical or radiological surveillance, or to avoid virilisation due to 5-alpha reductase deficiency in a 46,XY girl with a stable female gender identity.45 Pathological examination of DSD gonads requires specific expertise. For example, the differentiation between benign germ cell abnormalities, such as delayed maturation and (pre)malignant development of germ cells, is crucial for clinical management but can be very troublesome.46 Centralised pathological examination of gonadal biopsy and gonadectomy samples in one centre, or a restricted number of centres, on a national scale can help to overcome the problem of non-uniform classification and has proven feasible in the Netherlands and Belgium. We therefore believe that uniform assessment and classification of gonadal differentiation patterns should also be addressed in guidelines on DSD management.International databases of gonadal tissues are crucial for learning more about the risk of malignancy in different forms of DSD, but they are only reliable if uniform criteria for histological classification are strictly applied.46 These criteria could be incorporated in many existing networks such as the I-DSD consortium, the Disorders of Sex Development Translational Research Network, the European Reference Network on Urogenital Diseases (eUROGEN), the EndoERN and COST actions.15–17 47Communication at the transition from paediatric to adult carePaediatric and adult teams need to collaborate closely to facilitate a well-organised transition from paediatric to adult specialist care.15 48–50 Both teams need to exchange information optimally and should consider transition as a longitudinal process rather than a fixed moment in time.

Age-appropriate information is key at all ages, and an overview of topics to be discussed at each stage is described by Cools et al.15 Table 1 shows an example of how transition can be organised.View this table:Table 1 Example of transition table as used in the DSD clinic of the Erasmus Medical CenterPsychological support and the continued provision of information remains important for individuals with a DSD at all ages.15 22 In addition to the information given by the DSD team members, families and patients can benefit from resources such as support groups and information available on the internet.47 Information available online should be checked for accuracy and completeness when referring patients and parents to internet sites.Recommendations for future actionsMost guidelines and articles on the diagnosis and management of DSD are aimed at specialists and are only published in specialist journals or on websites for endocrinologists, urologists or geneticists. Yet there is a need for guidelines directed towards first-line and second-line healthcare workers that summarise the recommendations about the first crucial steps in the management of DSD. These should be published in widely available general medical journals and online, along with a national list of DSD centres. Furthermore, DSD (expert) centres should provide continuous education to all those who may be involved in the identification of individuals with a DSD in order to enable these healthcare professionals to recognise atypical genitalia, to promptly refer individuals who have a DSD and to inform the patient and parents about this and subsequent diagnostic procedures.As DSD continues to be a rare condition, it will take time to evaluate the effects of having such a guideline on the preparedness of first-line and second-line healthcare workers to recognise DSD conditions.

One way to evaluate this might be the development and use of questionnaires asking patients, carers and families and referring physicians how satisfied they were with the initial medical consultation and referral and what could be improved. A helpful addition to existing international databases that collect information on genetic variations would be a list of centres that offer suitable functional studies for certain genes, ideally covering the most frequently mutated genes (at minimum).Patient organisations can also play an important role in informing patients about newly available diagnostic or therapeutic strategies and options, and their influence and specific role has now been recognised and discussed in several publications.17 47 However, it should be kept in mind that these organisations do not represent all patients, as a substantial number of patients and parents are not member of such an organisation.Professionals have to provide optimal medical care based on well-established evidence, or at least on broad consensus. Yet not everything can be regulated by recommendations and guidelines. Options, ideas and wishes should be openly discussed between professionals, patients and families within their confidential relationship.

This will enable highly individualised holistic care tailored to the patient’s needs and expectations. Once they are well-informed of all available options, parents and/or patients can choose what they consider the optimal care for their children or themselves.15 16ConclusionThe Dutch-Flemish guideline uniquely addresses some topics that are under-represented in the literature, thus adding some key aspects to those addressed in recent consensus papers and guidelines.15–17 33 47As more children with a DSD are now being identified prenatally, and the literature on prenatal diagnosis of DSD remains scarce,20 21 we propose a prenatal diagnostic algorithm and emphasise the importance of having a prenatal specialist involved in or collaborating with DSD (expert) centres.We also stress that good communication between all involved parties is essential. Professionals should be well informed about protocols and communication. Collaboration between centres is necessary to optimise aspects of care such as uniform interpretation of gonadal pathology and functional testing of class 3 variants found by genetic testing.

Guidelines can provide a framework within which individualised patient care should be discussed with all stakeholders.AcknowledgmentsThe authors would like to thank the colleagues of the DSD teams for their input in and critical reading of the Dutch-Flemish guideline. Amsterdam University Center (AMC and VU), Maastricht University Medical Center, Erasmus Medical Center Rotterdam, Radboud University Medical Center Nijmegen, University Medical Center Groningen, University Medical Center Utrecht, Ghent University Hospital. The authors would like to thank Kate McIntyre for editing the revised manuscript and Tom de Vries Lentsch for providing the figures as a PDF. Three of the authors of this publication are members of the European Reference Network for rare endocrine diseases—Project ID 739543.IntroductionEndometrial cancer is the most common gynaecological malignancy in the developed world.1 Its incidence has risen over the last two decades as a consequence of the ageing population, fewer hysterectomies for benign disease and the obesity epidemic.

In the USA, it is estimated that women have a 1 in 35 lifetime risk of endometrial cancer, and in contrast to cancers of most other sites, cancer-specific mortality has risen by approximately 2% every year since 2008 related to the rapidly rising incidence.2Endometrial cancer has traditionally been classified into type I and type II based on morphology.3 The more common subtype, type I, is mostly comprised of endometrioid tumours and is oestrogen-driven, arises from a hyperplastic endometrium, presents at an early stage and has an excellent 5 year survival rate.4 By contrast, type II includes non-endometrioid tumours, specifically serous, carcinosarcoma and clear cell subtypes, which are biologically aggressive tumours with a poor prognosis that are often diagnosed at an advanced stage.5 Recent efforts have focused on a molecular classification system for more accurate categorisation of endometrial tumours into four groups with distinct prognostic profiles.6 7The majority of endometrial cancers arise through the interplay of familial, genetic and lifestyle factors. Two inherited cancer predisposition syndromes, Lynch syndrome and the much rarer Cowden syndrome, substantially increase the lifetime risk of endometrial cancer, but these only account for around 3–5% of cases.8–10 Having first or second degree relative(s) with endometrial or colorectal cancer increases endometrial cancer risk, although a large European twin study failed to demonstrate a strong heritable link.11 The authors failed to show that there was greater concordance in monozygotic than dizygotic twins, but the study was based on relatively small numbers of endometrial cancers. Lu and colleagues reported an association between common single nucleotide polymorphisms (SNPs) and endometrial cancer risk, revealing the potential role of SNPs in explaining part of the risk in both the familial and general populations.12 Thus far, many SNPs have been reported to modify susceptibility to endometrial cancer. However, much of this work predated genome wide association studies and is of variable quality.

Understanding genetic predisposition to endometrial cancer could facilitate personalised risk assessment with a view to targeted prevention and screening interventions.13 This emerged as the most important unanswered research question in endometrial cancer according to patients, carers and healthcare professionals in our recently completed James Lind Womb Cancer Alliance Priority Setting Partnership.14 It would be particularly useful for non-endometrioid endometrial cancers, for which advancing age is so far the only predictor.15We therefore conducted a comprehensive systematic review of the literature to provide an overview of the relationship between SNPs and endometrial cancer risk. We compiled a list of the most robust endometrial cancer-associated SNPs. We assessed the applicability of this panel of SNPs with a theoretical polygenic risk score (PRS) calculation. We also critically appraised the meta-analyses investigating the most frequently reported SNPs in MDM2.

Finally, we described all SNPs reported within genes and pathways that are likely involved in endometrial carcinogenesis and metastasis.MethodsOur systematic review follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) collaboration 2009 recommendations. The registered protocol is available through PROSPERO (CRD42018091907).16Search strategyWe searched Embase, MEDLINE and Cumulative Index to Nursing and Allied Health Literature (CINAHL) databases via the Healthcare Databases Advanced Search (HDAS) platform, from 2007 to 2018, to identify studies reporting associations between polymorphisms and endometrial cancer risk. Key words including MeSH (Medical Subject Heading) terms and free-text words were searched in both titles and abstracts. The following terms were used.

€œendomet*”,“uter*”, “womb”, “cancer(s)”, “neoplasm(s)”, “endometrium tumour”, “carcinoma”, “adenosarcoma”, “clear cell carcinoma”, “carcinosarcoma”, “SNP”, “single nucleotide polymorphism”, “GWAS”, and “genome-wide association study/ies”. No other restrictions were applied. The search was repeated with time restrictions between 2018 and June 2019 to capture any recent publications.Eligibility criteriaStudies were selected for full-text evaluation if they were primary articles investigating a relationship between endometrial cancer and SNPs. Study outcome was either the increased or decreased risk of endometrial cancer relative to controls reported as an odds ratio (OR) with corresponding 95% confidence intervals (95% CIs).Study selectionThree independent reviewers screened all articles uploaded to a screening spreadsheet developed by Helena VonVille.17 Disagreements were resolved by discussion.

Chronbach’s α score was calculated between reviewers and indicated high consistency at 0.92. Case–control, prospective and retrospective studies, genome-wide association studies (GWAS), and both discovery and validation studies were selected for full-text evaluation. Non-English articles, editorials, conference abstracts and proceedings, letters and correspondence, case reports and review articles were excluded.Candidate-gene studies with at least 100 women and GWAS with at least 1000 women in the case arm were selected to ensure reliability of the results, as explained by Spencer et al.18 To construct a panel of up to 30 SNPs with the strongest evidence of association, those with the strongest p values were selected. For the purpose of an SNP panel, articles utilising broad European or multi-ethnic cohorts were selected.

Where overlapping populations were identified, the most comprehensive study was included.Data extraction and synthesisFor each study, the following data were extracted. SNP ID, nearby gene(s)/chromosome location, OR (95% CI), p value, minor or effect allele frequency (MAF/EAF), EA (effect allele) and OA (other allele), adjustment, ethnicity and ancestry, number of cases and controls, endometrial cancer type, and study type including discovery or validation study and meta-analysis. For risk estimates, a preference towards most adjusted results was applied. For candidate-gene studies, a standard p value of<0.05 was applied and for GWAS a p value of <5×10-8, indicating genome-wide significance, was accepted as statistically significant.

However, due to the limited number of SNPs with p values reaching genome-wide significance, this threshold was then lowered to <1×10-5, allowing for marginally significant SNPs to be included. As shown by Mavaddat et al, for breast cancer, SNPs that fall below genome-wide significance may still be useful for generating a PRS and improving the models.19We estimated the potential value of a PRS based on the most significant SNPs by comparing the predicted risk for a woman with a risk score in the top 1% of the distribution to the mean predicted risk. Per-allele ORs and MAFs were taken from the publications and standard errors (SEs) for the lnORs were derived from published 95% CIs. The PRS was assumed to have a Normal distribution, with mean 2∑βipI and SE, σ, equal to √2∑βi2pI(1−pi), according to the binomial distribution, where the summation is over all SNPs in the risk score.

Hence the relative risk (RR) comparing the top 1% of the distribution to the mean is given by exp(Z0.01σ), where Z is the inverse of the standard normal cumulative distribution.ResultsThe flow chart of study selection is illustrated in figure 1. In total, 453 text articles were evaluated and, of those, 149 articles met our inclusion criteria. One study was excluded from table 1, for having an Asian-only population, as this would make it harder to compare with the rest of the results which were all either multi-ethnic or Caucasian cohorts, as stated in our inclusion criteria for the SNP panel.20 Any SNPs without 95% CIs were also excluded from any downstream analysis. Additionally, SNPs in linkage disequilibrium (r2 >0.2) with each other were examined, and of those in linkage disequilibrium, the SNP with strongest association was reported.

Per allele ORs were used unless stated otherwise.View this table:Table 1 List of top SNPs most likely to contribute to endometrial cancer risk identified through systematic review of recent literature21–25Study selection flow diagram. *Reasons. Irrelevant articles, articles focusing on other conditions, non-GWAS/candidate-gene study related articles, technical and duplicate articles. GWAS, genome-wide association study.

Adapted from. Moher D, Liberati A, Tetzlaff J, Altman DG, The PRISMA Group (2009). Preferred Reporting Items for Systematic Reviews and Meta-Analyses. The PRISMA Statement.

PLoS Med 6(6). E1000097. Doi:10.1371/journal.pmed1000097." data-icon-position data-hide-link-title="0">Figure 1 Study selection flow diagram. *Reasons.

Irrelevant articles, articles focusing on other conditions, non-GWAS/candidate-gene study related articles, technical and duplicate articles. GWAS, genome-wide association study. Adapted from. Moher D, Liberati A, Tetzlaff J, Altman DG, The PRISMA Group (2009).

Preferred Reporting Items for Systematic Reviews and Meta-Analyses. The PRISMA Statement. PLoS Med 6(6). E1000097.

Doi:10.1371/journal.pmed1000097.Top SNPs associated with endometrial cancer riskFollowing careful interpretation of the data, 24 independent SNPs with the lowest p values that showed the strongest association with endometrial cancer were obtained (table 1).21–25 These SNPs are located in or around genes coding for transcription factors, cell growth and apoptosis regulators, and enzymes involved in the steroidogenesis pathway. All the SNPs presented here were reported on the basis of a GWAS or in one case, an exome-wide association study, and hence no SNPs from candidate-gene studies made it to the list. This is partly due to the nature of larger GWAS providing more comprehensive and powered results as opposed to candidate gene studies. Additionally, a vast majority of SNPs reported by candidate-gene studies were later refuted by large-scale GWAS such as in the case of TERT and MDM2 variants.26 27 The exception to this is the CYP19 gene, where candidate-gene studies reported an association between variants in this gene with endometrial cancer in both Asian and broad European populations, and this association was more recently confirmed by large-scale GWAS.21 28–30 Moreover, a recent article authored by O’Mara and colleagues reviewed the GWAS that identified most of the currently known SNPs associated with endometrial cancer.31Most of the studies represented in table 1 are GWAS and the majority of these involved broad European populations.

Those having a multi-ethnic cohort also consisted primarily of broad European populations. Only four of the variants in table 1 are located in coding regions of a gene, or in regulatory flanking regions around the gene. Thus, most of these variants would not be expected to cause any functional effects on the gene or the resulting protein. An eQTL search using GTEx Portal showed that some of the SNPs are significantly associated (p<0.05) with modified transcription levels of the respective genes in various tissues such as prostate (rs11263761), thyroid (rs9668337), pituitary (rs2747716), breast mammary (rs882380) and testicular (rs2498794) tissue, as summarised in table 2.View this table:Table 2 List of eQTL hits for the selected panel of SNPsThe only variant for which there was an indication of a specific association with non-endometrioid endometrial cancer was rs148261157 near the BCL11A gene.

The A allele of this SNP had a moderately higher association in the non-endometrioid arm (OR 1.64, 95% CI 1.32 to 2.04. P=9.6×10-6) compared with the endometrioid arm (OR 1.25, 95% CI 1.14 to 1.38. P=4.7×10-6).21Oestrogen receptors α and β encoded by ESR1 and ESR2, respectively, have been extensively studied due to the assumed role of oestrogens in the development of endometrial cancer. O’Mara et al reported a lead SNP (rs79575945) in the ESR1 region that was associated with endometrial cancer (p=1.86×10-5).24 However, this SNP did not reach genome-wide significance in a more recent larger GWAS.21 No statistically significant associations have been reported between endometrial cancer and SNPs in the ESR2 gene region.AKT is an oncogene linked to endometrial carcinogenesis.

It is involved in the PI3K/AKT/mTOR pro-proliferative signalling pathway to inactivate apoptosis and allow cell survival. The A allele of rs2494737 and G allele of rs2498796 were reported to be associated with increased and decreased risk of endometrial cancer in 2016, respectively.22 30 However, this association was not replicated in a larger GWAS in 2018.21 Nevertheless, given the previous strong indications, and biological basis that could explain endometrial carcinogenesis, we decided to include an AKT1 variant (rs2498794) in our results.PTEN is a multi-functional tumour suppressor gene that regulates the AKT/PKB signalling pathway and is commonly mutated in many cancers including endometrial cancer.32 Loss-of-function germline mutations in PTEN are responsible for Cowden syndrome, which exerts a lifetime risk of endometrial cancer of up to 28%.9 Lacey and colleagues studied SNPs in the PTEN gene region. However, none showed significant differences in frequency between 447 endometrial cancer cases and 439 controls of European ancestry.33KRAS mutations are known to be present in endometrial cancer. These can be activated by high levels of KLF5 (transcriptional activator).

Three SNPs have been identified in or around KLF5 that are associated with endometrial cancer. The G allele of rs11841589 (OR 1.15, 95% CI 1.11 to 1.21. P=4.83×10-11), the A allele of rs9600103 (OR 1.23, 95% CI 1.16 to 1.30. P=3.76×10-12) and C allele of rs7981863 (OR 1.16, 95% CI 1.12 to 1.20.

P=2.70×10-17) have all been found to be associated with an increased likelihood of endometrial cancer in large European cohorts.21 30 34 It is worth noting that these SNPs are not independent, and hence they quite possibly tag the same causal variant.The MYC family of proto-oncogenes encode transcription factors that regulate cell proliferation, which can contribute to cancer development if dysregulated. The recent GWAS by O’Mara et al reported three SNPs within the MYC region that reached genome-wide significance with conditional p values reaching at least 5×10–8.35To test the utility of these SNPs as predictive markers, we devised a theoretical PRS calculation using the log ORs and EAFs per SNP from the published data. The results were very encouraging with an RR of 3.16 for the top 1% versus the mean, using all the top SNPs presented in table 1 and 2.09 when using only the SNPs that reached genome-wide significance (including AKT1).Controversy surrounding MDM2 variant SNP309MDM2 negatively regulates tumour suppressor gene TP53, and as such, has been extensively studied in relation to its potential role in predisposition to endometrial cancer. Our search identified six original studies of the association between MDM2 SNP rs2279744 (also referred to as SNP309) and endometrial cancer, all of which found a statistically significant increased risk per copy of the G allele.

Two more original studies were identified through our full-text evaluation. However, these were not included here as they did not meet our inclusion criteria—one due to small sample size, the other due to studying rs2279744 status dependent on another SNP.36 37 Even so, the two studies were described in multiple meta-analyses that are listed in table 3. Different permutations of these eight original studies appear in at least eight published meta-analyses. However, even the largest meta-analysis contained <2000 cases (table 3)38View this table:Table 3 Characteristics of studies that examined MDM2 SNP rs2279744In comparison, a GWAS including nearly 13 000 cases found no evidence of an association with OR and corresponding 95% CI of 1.00 (0.97 to 1.03) and a p value of 0.93 (personal communication).21 Nevertheless, we cannot completely rule out a role for MDM2 variants in endometrial cancer predisposition as the candidate-gene studies reported larger effects in Asians, whereas the GWAS primarily contained participants of European ancestry.

There is also some suggestion that the SNP309 variant is in linkage disequilibrium with another variant, SNP285, which confers an opposite effect.It is worth noting that the SNP285C/SNP309G haplotype frequency was observed in up to 8% of Europeans, thus requiring correction for the confounding effect of SNP285C in European studies.39 However, aside from one study conducted by Knappskog et al, no other study including the meta-analyses corrected for the confounding effect of SNP285.40 Among the studies presented in table 3, Knappskog et al (2012) reported that after correcting for SNP285, the OR for association of this haplotype with endometrial cancer was much lower, though still significant. Unfortunately, the meta-analyses which synthesised Knappskog et al (2012), as part of their analysis, did not correct for SNP285C in the European-based studies they included.38 41 42 It is also concerning that two meta-analyses using the same primary articles failed to report the same result, in two instances.38 42–44DiscussionThis article represents the most comprehensive systematic review to date, regarding critical appraisal of the available evidence of common low-penetrance variants implicated in predisposition to endometrial cancer. We have identified the most robust SNPs in the context of endometrial cancer risk. Of those, only 19 were significant at genome-wide level and a further five were considered marginally significant.

The largest GWAS conducted in this field was the discovery- and meta-GWAS by O’Mara et al, which utilised 12 096 cases and 108 979 controls.21 Despite the inclusion of all published GWAS and around 5000 newly genotyped cases, the total number did not reach anywhere near what is currently available for other common cancers such as breast cancer. For instance, BCAC (Breast Cancer Association Consortium) stands at well over 200 000 individuals with more than half being cases, and resulted in identification of ~170 SNPs in relation to breast cancer.19 45 A total of 313 SNPs including imputations were then used to derive a PRS for breast cancer.19 Therefore, further efforts should be directed to recruit more patients, with deep phenotypic clinical data to allow for relevant adjustments and subgroup analyses to be conducted for better precision.A recent pre-print study by Zhang and colleagues examined the polygenicity and potential for SNP-based risk prediction for 14 common cancers, including endometrial cancer, using available summary-level data from European-ancestry datasets.46 They estimated that there are just over 1000 independent endometrial cancer susceptibility SNPs, and that a PRS comprising all such SNPs would have an area under the receiver-operator curve of 0.64, similar to that predicted for ovarian cancer, but lower than that for the other cancers in the study. The modelling in the paper suggests that an endometrial cancer GWAS double the size of the current largest study would be able to identify susceptibility SNPs together explaining 40% of the genetic variance, but that in order to explain 75% of the genetic variance it would be necessary to have a GWAS comprising close to 150 000 cases and controls, far in excess of what is currently feasible.We found that the literature consists mainly of candidate-gene studies with small sample sizes, meta-analyses reporting conflicting results despite using the same set of primary articles, and multiple reports of significant SNPs that have not been validated by any larger GWAS. The candidate-gene studies were indeed the most useful and cheaper technique available until the mid to late 2000s.

However, a lack of reproducibility (particularly due to population stratification and reporting bias), uncertainty of reported associations, and considerably high false discovery rates make these studies much less appropriate in the post-GWAS era. Unlike the candidate-gene approach, GWAS do not require prior knowledge, selection of genes or SNPs, and provide vast amounts of data. Furthermore, both the genotyping process and data analysis phases have become cheaper, the latter particularly due to faster and open-access pre-phasing and imputation tools being made available.It is clear from table 1 that some SNPs were reported with wide 95% CI, which can be directly attributed to small sample sizes particularly when restricting the cases to non-endometrioid histology only, low EAF or poor imputation quality. Thus, these should be interpreted with caution.

Additionally, most of the SNPs reported by candidate-gene studies were not detected by the largest GWAS to date conducted by O’Mara et al.21 However, this does not necessarily mean that the possibility of those SNPs being relevant should be completely dismissed. Moreover, meta-analyses were attempted for other variants. However, these showed no statistically significant association and many presented with high heterogeneity between the respective studies (data not shown). Furthermore, as many studies utilised the same set of cases and/or controls, conducting a meta-analysis was not possible for a good number of SNPs.

It is therefore unequivocal that the literature is crowded with numerous small candidate-gene studies and conflicting data. This makes it particularly hard to detect novel SNPs and conduct meaningful meta-analyses.We found convincing evidence for 19 variants that indicated the strongest association with endometrial cancer, as shown in table 1. The associations between endometrial cancer and variants in or around HNF1B, CYP19A1, SOX4, MYC, KLF and EIF2AK found in earlier GWAS were then replicated in the latest and largest GWAS. These SNPs showed promising potential in a theoretical PRS we devised based on published data.

Using all 24 or genome-wide significant SNPs only, women with a PRS in the top 1% of the distribution would be predicted to have a risk of endometrial cancer 3.16 and 2.09 times higher than the mean risk, respectively.However, the importance of these variants and relevance of the proximate genes in a functional or biological context is challenging to evaluate. Long distance promoter regulation by enhancers may disguise the genuine target gene. In addition, enhancers often do not loop to the nearest gene, further complicating the relevance of nearby gene(s) to a GWAS hit. In order to elucidate biologically relevant candidate target genes in endometrial cancer, O’Mara et al looked into promoter-associated chromatin looping using a modern HiChIP approach.47 The authors utilised normal and tumoural endometrial cell lines for this analysis which showed significant enrichment for endometrial cancer heritability, with 103 candidate target genes identified across the 13 risk loci identified by the largest ECAC GWAS.

Notable genes identified here were CDKN2A and WT1, and their antisense counterparts. The former was reported to be nearby of rs1679014 and the latter of rs10835920, as shown in table 1. Moreover, of the 36 candidate target genes, 17 were found to be downregulated while 19 were upregulated in endometrial tumours.The authors also investigated overlap between the 13 endometrial cancer risk loci and top eQTL variants for each target gene.47 In whole blood, of the two particular lead SNPs, rs8822380 at 17q21.32 was a top eQTL for SNX11 and HOXB2, whereas rs937213 at 15q15.1 was a top eQTL for SRP14. In endometrial tumour, rs7579014 at 2p16.1 was found to be a top eQTL for BCL11A.

This is particularly interesting because BCL11A was the only nearby/candidate gene that had a GWAS association reported in both endometrioid and non-endometrioid subtypes. The study looked at protein–protein interactions between endometrial cancer drivers and candidate target gene products. Significant interactions were observed with TP53 (most significant), AKT, PTEN, ESR1 and KRAS, among others. Finally, when 103 target candidate genes and 387 proteins were combined together, 462 pathways were found to be significantly enriched.

Many of these are related to gene regulation, cancer, obesity, insulinaemia and oestrogen exposure. This study clearly showed a potential biological relevance for some of the SNPs reported by ECAC GWAS in 2018.Most of the larger included studies used cohorts primarily composed of women of broad European descent. Hence, there are negligible data available for other ethnicities, particularly African women. This is compounded by the lack of reference genotype data available for comparative analysis, making it harder for research to be conducted in ethnicities other than Europeans.

This poses a problem for developing risk prediction models that are equally valuable and predictive across populations. Thus, our results also are of limited applicability to non-European populations.Furthermore, considering that non-endometrioid cases comprise a small proportion (~20%) of all endometrial cancer cases, much larger cohort sizes are needed to detect any genuine signals for non-endometrioid tumours. Most of the evaluated studies looked at either overall/mixed endometrial cancer subtypes or endometrioid histology, and those that looked at variant associations with non-endometrioid histology were unlikely to have enough power to detect any signal with statistical significance. This is particularly concerning because non-endometrioid subtypes are biologically aggressive tumours with a much poorer prognosis that contribute disproportionately to mortality from endometrial cancer.

It is particularly important that attempts to improve early detection and prevention of endometrial cancer focus primarily on improving outcomes from these subtypes. It is also worth noting that, despite the current shift towards a molecular classification of endometrial cancer, most studies used the overarching classical Bokhman’s classification system, type I versus type II, or no histological classification system at all. Therefore, it is important to create and follow a standardised and comprehensive classification system for reporting tumour subtypes for future studies.This study compiled and presented available information for an extensively studied, yet unproven in large datasets, SNP309 variant in MDM2. Currently, there is no convincing evidence for an association between this variant and endometrial cancer risk.

Additionally, of all the studies, only one accounted for the opposing effect of a nearby variant SNP285 in their analyses. Thus, we conclude that until confirmed by a sufficiently large GWAS, this variant should not be considered significant in influencing the risk of endometrial cancer and therefore not included in a PRS. This is also true for the majority of the SNPs reported in candidate-gene studies, as the numbers fall far short of being able to detect genuine signals.This systematic review presents the most up-to-date evidence for endometrial cancer susceptibility variants, emphasising the need for further large-scale studies to identify more variants of importance, and validation of these associations. Until data from larger and more diverse cohorts are available, the top 24 SNPs presented here are the most robust common genetic variants that affect endometrial cancer risk.

The multiplicative effects of these SNPs could be used in a PRS to allow personalised risk prediction models to be developed for targeted screening and prevention interventions for women at greatest risk of endometrial cancer..

Vibramycin cream

In early August, there was a lot of hubbub around a study that purportedly showed that wearing a neck gaiter, the sleeve-like face covering popular especially among runners, might be worse at stemming the spread of COVID-19 than not wearing a mask at all vibramycin cream. Headlines popped up spreading the news, sparking conversations far and wide and forcing many to reconsider their preferred style of face mask. A Washington Post story said “some cotton vibramycin cream cloth masks are about as effective as surgical masks, while thin polyester spandex gaiters may be worse than going maskless.” A Forbes article, referring to neck gaiters, said the study “found that one type of face covering might actually be doing more harm than good.” But the study didn’t show that, nor was it designed to. It was actually a test on how to test masks inexpensively, not to determine which one was most effective. The researchers set up a green laser beam in a dark room.

A masked subject was then vibramycin cream asked to speak so that the droplets from the speaker’s mouth showed up in the green beam. The whole process was video recorded on a cell phone, after which researchers calculated the number of droplets that showed up. The process was repeated 10 times for each mask (14 in total, one of which was a neck gaiter) and the setup cost less than $200. What was meant as a study on the pricing and efficacy of a test turned into, at least in vibramycin cream some journalistic circles, a definitive nail-in-the-coffin for gaiters. Days after the initial reports that neck gaiters might not only be useless but maybe even harmful, a new round of new reports came out saying that those initial reports were overblown and misleading.

The authors vibramycin cream of the study even held a press conference where they emphasized that their study was never meant to test the effectiveness of masks. They only tested one gaiter-style mask, which says nothing about that style of mask in general. The combination of reporting on the actual findings of the study and the direct comments from the authors seems to have abated the anti-neck gaiter fervor. But all of this this—or most of it, anyway—likely vibramycin cream could have been prevented. You could make the argument that it’s not a scientist’s job to worrying about how their science might be interpreted.

It’s their job to do the research and publish it in a scientific manuscript. Leave the communicating for someone vibramycin cream else. But that’s not how the spread of information works. Fewer and fewer newsrooms have staffers with scientific backgrounds, or who are dedicated to scientific reporting. To be vibramycin cream clear, journalists don’t need to be scientists to understand science, but reporting on science does require a certain amount of expertise.

When newsrooms ask reporters to cover more and more topic areas and this specialization decreases, an attention to detail is sometimes lost. So, the onus to help journalists (and frankly, all nonscientists) get vibramycin cream the facts straight falls to the scientists doing the science. That’s where science communication training comes in. Science communication, or scicomm as it’s known colloquially, is not a core part of coursework in a majority of degree-granting science programs at the undergraduate and graduate levels. This trend is slowly changing vibramycin cream as more institutions incorporate scicomm into their curriculums.

Outside of academia, nonprofits and scientific societies are taking up the mantle. I work for the American Geophysical Union (AGU), a society for Earth and space scientists, in the Sharing Science program, where we teach scientists to communicate with nonscientists through courses, workshops, webinars and other trainings. Aside from the AGU, there is the American Association for the Advancement of Science (AAAS), the Stony Brook–affiliated Alan Alda Center for Communicating Science and the science storytelling organization The Story Collider, to name just vibramycin cream to name a few. We teach the so-called “soft skills” that the ivory tower of science has shunned for so long but that are so necessary in effectively communicating. One thing we stress is “know your audience.” Scientists must think about how their science will be perceived, no matter how relevant or not vibramycin cream it might be to the broader public.

Science does not exist in a vacuum. It never has. But especially now, and especially with anything related to COVID-19, scientists much be hypervigilant when communicating results and try, to the best of their vibramycin cream abilities, to account for as many interpretations as possible. Yes, it is onerous, especially on top of the multitude of other responsibilities that come with being a scientist, but it is necessary. The traditional ways in which scientists communicate their results (i.e., scientific manuscripts) are not going away anytime soon.

However, and while it may be an unfair ask, scientists must not only be able to communicate their science to their peers vibramycin cream. They must always think about nonscience audiences as the lines between science and “the public” continue to blur. Training scientists to effectively communicate to, or at least think about, diverse audiences is a necessary part of science..

In early August, low cost vibramycin there was a lot of hubbub around a study that purportedly showed that wearing a neck gaiter, the sleeve-like face covering popular especially among runners, might be worse at stemming the spread of COVID-19 than not wearing a mask at all. Headlines popped up spreading the news, sparking conversations far and wide and forcing many to reconsider their preferred style of face mask. A Washington Post low cost vibramycin story said “some cotton cloth masks are about as effective as surgical masks, while thin polyester spandex gaiters may be worse than going maskless.” A Forbes article, referring to neck gaiters, said the study “found that one type of face covering might actually be doing more harm than good.” But the study didn’t show that, nor was it designed to. It was actually a test on how to test masks inexpensively, not to determine which one was most effective. The researchers set up a green laser beam in a dark room.

A masked subject was then asked to speak so that the droplets from the speaker’s mouth showed up in low cost vibramycin the green beam. The whole process was video recorded on a cell phone, after which researchers calculated the number of droplets that showed up. The process was repeated 10 times for each mask (14 in total, one of which was a neck gaiter) and the setup cost less than $200. What was meant as a study on the pricing and efficacy of a test low cost vibramycin turned into, at least in some journalistic circles, a definitive nail-in-the-coffin for gaiters. Days after the initial reports that neck gaiters might not only be useless but maybe even harmful, a new round of new reports came out saying that those initial reports were overblown and misleading.

The authors of the study even held a press conference where they emphasized low cost vibramycin that their study was never meant to test the effectiveness of masks. They only tested one gaiter-style mask, which says nothing about that style of mask in general. The combination of reporting on the actual findings of the study and the direct comments from the authors seems to have abated the anti-neck gaiter fervor. But all of this this—or most of low cost vibramycin it, anyway—likely could have been prevented. You could make the argument that it’s not a scientist’s job to worrying about how their science might be interpreted.

It’s their job to do the research and publish it in a scientific manuscript. Leave the communicating for someone else low cost vibramycin. But that’s not how the spread of information works. Fewer and fewer newsrooms have staffers with scientific backgrounds, or who are dedicated to scientific reporting. To be clear, journalists don’t need to be scientists to understand low cost vibramycin science, but reporting on science does require a certain amount of expertise.

When newsrooms ask reporters to cover more and more topic areas and this specialization decreases, an attention to detail is sometimes lost. So, the onus to help journalists (and frankly, all low cost vibramycin nonscientists) get the facts straight falls to the scientists doing the science. That’s where science communication training comes in. Science communication, or scicomm as it’s known colloquially, is not a core part of coursework in a majority of degree-granting science programs at the undergraduate and graduate levels. This trend is slowly changing as low cost vibramycin more institutions incorporate scicomm into their curriculums.

Outside of academia, nonprofits and scientific societies are taking up the mantle. I work for the American Geophysical Union (AGU), a society for Earth and space scientists, in the Sharing Science program, where we teach scientists to communicate with nonscientists through courses, workshops, webinars and other trainings. Aside from the AGU, there is the American low cost vibramycin Association for the Advancement of Science (AAAS), the Stony Brook–affiliated Alan Alda Center for Communicating Science and the science storytelling organization The Story Collider, to name just to name a few. We teach the so-called “soft skills” that the ivory tower of science has shunned for so long but that are so necessary in effectively communicating. One thing we stress low cost vibramycin is “know your audience.” Scientists must think about how their science will be perceived, no matter how relevant or not it might be to the broader public.

Science does not exist in a vacuum. It never has. But especially now, and especially with anything related to COVID-19, scientists much be hypervigilant when low cost vibramycin communicating results and try, to the best of their abilities, to account for as many interpretations as possible. Yes, it is onerous, especially on top of the multitude of other responsibilities that come with being a scientist, but it is necessary. The traditional ways in which scientists communicate their results (i.e., scientific manuscripts) are not going away anytime soon.

However, and while it may be an unfair ask, scientists must not only be able low cost vibramycin to communicate their science to their peers. They must always think about nonscience audiences as the lines between science and “the public” continue to blur. Training scientists to effectively communicate to, or at least think about, diverse audiences is a necessary part of science..

Generic vibramycin online for sale

You may be hearing about how virtual care, often described as telehealth or telemedicine, is beneficial during COVID-19 and how health systems are offering virtual generic vibramycin online for sale access like never before. There’s a reason for that, too. For the past few weeks I’ve seen Facebook posts daily from former nursing colleagues in metro Detroit, one of the hardest hit areas in the country, as they provide front-line care to patients with COVID-19.

It makes generic vibramycin online for sale me very proud to call these nurses my friends. As a former emergency department nurse, I recall the feeling of satisfaction knowing that I’ve helped someone on the worst day of their life. One of the best parts of being a nurse is knowing you matter to the only person in health care that truly matters.

The patient generic vibramycin online for sale. Several years ago I made the difficult decision to no longer perform bedside nursing and become a nurse administrator. The biggest loss from my transition is the feeling that what I do matters to the patient.

COVID-19 has forced a lot of us to rethink the generic vibramycin online for sale role we play in health care and what the real priority should be. Things that were top priorities three months ago have been rightfully cast aside to either care for patients in a pandemic or prepare for the unknown future of, “When is our turn?. € For me, COVID-19 has reignited the feeling that what I do matters as virtual care has become a powerful tool on the forefront of care during this crisis.

It has also shown that many of the powerful rules and regulations that limit virtual generic vibramycin online for sale care are not needed and should be discarded permanently. When I became the director of virtual care at our organization in 2015 I knew nothing about telehealth. Sure, I had seen a stroke robot in some Emergency Departments, and I had some friends that told me their insurance company lets them FaceTime a doctor for free (spoiler alert.

It’s not generic vibramycin online for sale FaceTime). I was tech-savvy from a consumer perspective and a tech novice from an IT perspective. Nevertheless, my team and I spent the next few years learning as we built one of the higher volume virtual care networks in the state of Michigan.

We discovered a lot generic vibramycin online for sale of barriers that keep virtual care from actually making the lives of patients and providers better and we also became experts in working around those barriers. But, there were two obstacles that we could not overcome. Government regulation and insurance provider willingness to cover virtual visits.

These two barriers effectively cripple most legitimate attempts to provide value-added direct-to-consumer virtual care, which I define as using virtual care technologies to provide care outside of our brick-and-mortar facilities, most commonly generic vibramycin online for sale in the patient home. The need to social distance, cancel appointments, close provider offices, keep from overloading emergency departments and urgent cares and shelter in place created instant demand for direct-to-consumer virtual care. In all honesty, I’ve always considered direct-to-consumer virtual care to be the flashy, must-have holiday gift of the year that organizations are convinced will be the way of the future.

If a health system wants to provide on-demand access to patients for generic vibramycin online for sale low-complexity acute conditions, they will easily find plenty of vendors that will sell them their app and their doctors and put the health system’s logo on it. What a health system will struggle with is to find is enough patient demand to cover the high cost. Remember my friends from earlier that told me about the app their insurance gave them?.

Nearly all of them followed that up by telling me they’ve never actually used generic vibramycin online for sale it. I am fortunate that I work for an organization that understands this and instead focuses on how can we provide care that our patients actually want and need from the doctors they want to see. Ironically, this fiscal year we had a corporate top priority around direct-to-consumer virtual care.

We wanted to generic vibramycin online for sale expand what we thought were some successful pilots and perform 500 direct-to-consumer visits. This year has been one of the hardest of my leadership career because, frankly, up until a month ago I was about to fail on this top priority. With only four months left, we were only about halfway there.

The biggest problem we generic vibramycin online for sale ran into was that every great idea a physician brought to me was instantly dead in the water because practically no insurance company would pay for it. There are (prior to COVID-19) a plethora of rules around virtual care billing but the simplest way to summarize it is that most virtual care will only be paid if it happens in a rural location and inside of a health care facility. It is extremely limited what will be paid for in the patient home and most of it is so specific that the average patient isn’t eligible to get any in-home virtual care.

Therefore, most good medical uses for direct-to-consumer care would be asking the patient to pay cash or the physician to forgo reimbursement for a visit that would be covered if it happened in generic vibramycin online for sale office. Add to that the massive capital and operating expenses it takes to build a virtual care network and you can see why these programs don’t exist. A month ago I was skeptical we’d have a robust direct-to-consumer program any time soon and then COVID-19 hit.

When COVID-19 started to spread rapidly in the United States, regulations and reimbursement rules were being stripped generic vibramycin online for sale daily. The first change that had major impact is when the Centers for Medicare and Medicaid Services (CMS) announced that they would temporarily begin reimbursing for virtual visits conducted in the patient’s home for COVID-19 and non-COVID related visits. We were already frantically designing a virtual program to handle the wave of COVID-19 screening visits that were overloading our emergency departments and urgent cares.

We were having plenty of discussions around reimbursement for this generic vibramycin online for sale clinic. Do we attempt to bill insurances knowing they will likely deny, do we do a cash clinic model or do we do this as a community benefit and eat the cost?. The CMS waiver gave us hope that we would be compensated for diverting patients away from reimbursed visits to a virtual visit that is more convenient for the patient and aligns with the concept of social distancing.

Realistically we generic vibramycin online for sale don’t know if we will be paid for any of this. We are holding all of the bills for at least 90 days while the industry sorts out the rules. I was excited by the reimbursement announcement because I knew we had eliminated one of the biggest direct-to-consumer virtual care barriers.

However, I was quickly brought back to reality when I was reminded that HIPAA (Health Insurance Portability and Accountability Act) still generic vibramycin online for sale existed. I had this crazy idea that during a pandemic we should make it as easy as possible for people to receive virtual care and that the best way to do that was to meet the patient on the device they are most comfortable with and the application (FaceTime, Facebook, Skype, etc.) that they use every day. The problem is nearly every app the consumer uses on a daily basis is banned by HIPAA because “it’s not secure.” I’m not quite sure what a hacker stands to gain by listening into to my doctor and me talk about how my kids yet again gave me strep throat but apparently the concern is great enough to stifle the entire industry.

Sure, not every health care discussion is as low-key as strep throat and a patient may want generic vibramycin online for sale to protect certain topics from being discussed over a “non-secure” app but why not let the patient decide through informed consent?. Regulators could also abandon this all-or-nothing approach and lighten regulations surrounding specific health conditions. The idea that regulations change based on medical situation is not new.

For example, in my home state of generic vibramycin online for sale Michigan, adolescents are essentially considered emancipated if it involves sexual health, mental health or substance abuse. Never mind that this same information is freely given over the phone by every office around the country daily without issue, but I digress. While my job is to innovate new pathways for care, our lawyer’s job is to protect the organization and he, along with IT security, rightfully shot down my consumer applications idea.

A few days later I legitimately screamed out loud in joy when generic vibramycin online for sale the Department of Health and Human Services announced that it would use discretion on enforcing HIPAA compliance rules and specifically allowed for use of consumer applications. The elimination of billing restrictions and HIPAA regulations changed what is possible for health care organizations to offer virtually. Unfortunately both changes are listed as temporary and will likely be removed when the pandemic ends.

Six days after the HIPAA changes were announced, we launched a centralized generic vibramycin online for sale virtual clinic for any patient that wanted a direct-to-consumer video visit to be screened by a provider for COVID-19. It allows patients to call in without a referral and most patients are on-screen within five minutes of clicking the link we text them. They don’t have to download an app, create an account or even be an established patient of our health system.

It saw over 900 generic vibramycin online for sale patients in the first 12 days it was open. That is 900 real patients that received care from a physician or advanced practice provider without risking personal exposure and without going to an already overwhelmed ED or urgent care. To date, 70 percent of the patients seen by the virtual clinic did not meet CDC testing criteria for COVID-19.

I don’t believe we could have reached even half of these patients had the consumer application restrictions been generic vibramycin online for sale kept. A program like this almost certainly wouldn’t exist if not for the regulations being lifted and even if it did, it would have taken six to 12 months to navigate barriers and implement in normal times. Sure, the urgency of a pandemic helps but the impact of provider, patients, regulators and payors being on the same page is what fueled this fire.

During the virtual clinic’s first two weeks, my team turned its generic vibramycin online for sale attention to getting over 300 providers across 60+ offices virtual so they could see their patients at home. Imagine being an immunocompromised cancer patient right now and being asked to leave your home and be exposed to other people in order to see your oncologist. Direct-to-consumer virtual care is the best way to safely care for these patients and without these temporary waivers it wouldn’t be covered by insurance even if you did navigate the clunky apps that are HIPAA compliant.

Do we really think the immunocompromised cancer patient feels generic vibramycin online for sale any more comfortable every normal flu season?. Is it any more appropriate to ask them to risk exposure to the flu than it is to COVID-19?. And yet we deny them this access in normal times and it quite possibly will be stripped away from them when this crisis is over.

Now 300 to 400 patients per day in our health system are seen virtually by generic vibramycin online for sale their own primary care doctor or specialist for non-COVID related visits. Not a single one of these would have been reimbursed one month ago and I am highly skeptical I would have gotten approval to use the software that connects us to the patient. Lastly, recall that prior to COVID-19, our system had only found 250 total patients that direct-to-consumer care was value-added and wasn’t restricted by regulation or reimbursement.

COVID-19 has been a wake-up call to generic vibramycin online for sale the whole country and health care is no exception. It has put priorities in perspective and shined a light on what is truly value-added. For direct-to-consumer virtual care it has shown us what is possible when we get out of our own way.

If a regulation has to be removed to allow for care during a crisis then we must generic vibramycin online for sale question why it exists in the first place. HIPAA regulation cannot go back to its antiquated practices if we are truly going to shift the focus to patient wellness. CMS and private payors must embrace value-added direct-to-consumer virtual care and allow patients the access they deserve.

COVID-19 has forced this industry forward, we cannot allow it generic vibramycin online for sale to regress and be forgotten when this is over. Tom Wood is the director of trauma and virtual care for MidMichigan Health, a non-profit health system headquartered in Midland, Michigan, affiliated with Michigan Medicine, the health care division of the University of Michigan. The views and opinions expressed in this commentary are his own.When dealing with all of the aspects of diabetes, it’s easy to let your feel fall to the bottom of the list.

But daily care and evaluation is one of the best generic vibramycin online for sale ways to prevent foot complications. It’s important to identify your risk factors and take the proper steps in limiting your complications. Two of the biggest complications with diabetes are peripheral neuropathy and ulcer/amputation.

Symptoms of peripheral neuropathy include numbness, tingling generic vibramycin online for sale and/or burning in your feet and legs. You can slow the progression of developing neuropathy by making it a point to manage your blood sugars and keep them in the normal range. If you are experiencing these symptoms, it is important to establish and maintain a relationship with a podiatrist.

Your podiatrist can make sure things are looking generic vibramycin online for sale healthy and bring things to your attention to monitor and keep a close eye on. Open wounds or ulcers can develop secondary to trauma, pressure, diabetes, neuropathy or poor circulation. If ulcerations do develop, it’s extremely important to identify the cause and address it.

Ulcers can get worse quickly, so it’s necessary to seek immediate medical treatment if you find yourself or a loved one generic vibramycin online for sale dealing with this complication. Untreated ulcerations often lead to amputation and can be avoided if proper medical attention is sought right away. There are important things to remember when dealing with diabetic foot care.

It’s very important to inspect your feet daily, especially if you have generic vibramycin online for sale peripheral neuropathy. You may have a cut or a sore on your feet that you can’t feel, so your body doesn’t alarm you to check your feet. Be gentle when bathing your feet.

Moisturize your feet, but not between your toes. Do not treat calluses or corns on your own. Wear clean, dry socks.

Never walk barefoot, and consider socks and shoes made specifically for patients with diabetes.

For the past few low cost vibramycin weeks I’ve seen Facebook posts daily from former nursing colleagues in metro Detroit, one of the hardest hit areas in the country, as they provide front-line care to patients with COVID-19. It makes me very proud to call these nurses my friends. As a former emergency department nurse, I recall the feeling of satisfaction knowing that I’ve helped someone on the worst day of their life. One of the best parts of being a nurse low cost vibramycin is knowing you matter to the only person in health care that truly matters. The patient.

Several years ago I made the difficult decision to no longer perform bedside nursing and become a nurse administrator. The biggest loss from my low cost vibramycin transition is the feeling that what I do matters to the patient. COVID-19 has forced a lot of us to rethink the role we play in health care and what the real priority should be. Things that were top priorities three months ago have been rightfully cast aside to either care for patients in a pandemic or prepare for the unknown future of, “When is our turn?. € For me, COVID-19 has reignited the feeling that what I do matters as virtual care has become a powerful tool on the forefront of low cost vibramycin care during this crisis.

It has also shown that many of the powerful rules and regulations that limit virtual care are not needed and should be discarded permanently. When I became the director of virtual care at our organization in 2015 I knew nothing about telehealth. Sure, I had seen a stroke robot in some Emergency Departments, and low cost vibramycin I had some friends that told me their insurance company lets them FaceTime a doctor for free (spoiler alert. It’s not FaceTime). I was tech-savvy from a consumer perspective and a tech novice from an IT perspective.

Nevertheless, my team and I spent the next few years learning as we built one of the higher volume virtual care networks in the state of Michigan low cost vibramycin. We discovered a lot of barriers that keep virtual care from actually making the lives of patients and providers better and we also became experts in working around those barriers. But, there were two obstacles that we could not overcome. Government regulation and insurance provider willingness to cover low cost vibramycin virtual visits. These two barriers effectively cripple most legitimate attempts to provide value-added direct-to-consumer virtual care, which I define as using virtual care technologies to provide care outside of our brick-and-mortar facilities, most commonly in the patient home.

The need to social distance, cancel appointments, close provider offices, keep from overloading emergency departments and urgent cares and shelter in place created instant demand for direct-to-consumer virtual care. In all honesty, I’ve always considered direct-to-consumer virtual care to be the flashy, must-have holiday gift of the year that organizations are convinced will be the way of the low cost vibramycin future. If a health system wants to provide on-demand access to patients for low-complexity acute conditions, they will easily find plenty of vendors that will sell them their app and their doctors and put the health system’s logo on it. What a health system will struggle with is to find is enough patient demand to cover the high cost. Remember my friends from earlier that told me about the app low cost vibramycin their insurance gave them?.

Nearly all of them followed that up by telling me they’ve never actually used it. I am fortunate that I work for an organization that understands this and instead focuses on how can we provide care that our patients actually want and need from the doctors they want to see. Ironically, this fiscal year we had a corporate top priority around direct-to-consumer low cost vibramycin virtual care. We wanted to expand what we thought were some successful pilots and perform 500 direct-to-consumer visits. This year has been one of the hardest of my leadership career because, frankly, up until a month ago I was about to fail on this top priority.

With only four months left, we were low cost vibramycin only about halfway there. The biggest problem we ran into was that every great idea a physician brought to me was instantly dead in the water because practically no insurance company would pay for it. There are (prior to COVID-19) a plethora of rules around virtual care billing but the simplest way to summarize it is that most virtual care will only be paid if it happens in a rural location and inside of a health care facility. It is extremely limited what will be paid for in the low cost vibramycin patient home and most of it is so specific that the average patient isn’t eligible to get any in-home virtual care. Therefore, most good medical uses for direct-to-consumer care would be asking the patient to pay cash or the physician to forgo reimbursement for a visit that would be covered if it happened in office.

Add to that the massive capital and operating expenses it takes to build a virtual care network and you can see why these programs don’t exist. A month ago I was skeptical we’d have a low cost vibramycin robust direct-to-consumer program any time soon and then COVID-19 hit. When COVID-19 started to spread rapidly in the United States, regulations and reimbursement rules were being stripped daily. The first change that had major impact is when the Centers for Medicare and Medicaid Services (CMS) announced that they would temporarily begin reimbursing for virtual visits conducted in the patient’s home for COVID-19 and non-COVID related visits. We were already frantically designing a virtual program to handle the wave of COVID-19 screening visits that were overloading our emergency departments and urgent low cost vibramycin cares.

We were having plenty of discussions around reimbursement for this clinic. Do we attempt to bill insurances knowing they will likely deny, do we do a cash clinic model or do we do this as a community benefit and eat the cost?. The CMS waiver low cost vibramycin gave us hope that we would be compensated for diverting patients away from reimbursed visits to a virtual visit that is more convenient for the patient and aligns with the concept of social distancing. Realistically we don’t know if we will be paid for any of this. We are holding all of the bills for at least 90 days while the industry sorts out the rules.

I was excited low cost vibramycin by the reimbursement announcement because I knew we had eliminated one of the biggest direct-to-consumer virtual care barriers. However, I was quickly brought back to reality when I was reminded that HIPAA (Health Insurance Portability and Accountability Act) still existed. I had this crazy idea that during a pandemic we should make it as easy as possible for people to receive virtual care and that the best way to do that was to meet the patient on the device they are most comfortable with and the application (FaceTime, Facebook, Skype, etc.) that they use every day. The problem is nearly every app the consumer uses on a daily basis is banned by HIPAA because “it’s not secure.” I’m not quite sure what a hacker stands to gain by listening into to my low cost vibramycin doctor and me talk about how my kids yet again gave me strep throat but apparently the concern is great enough to stifle the entire industry. Sure, not every health care discussion is as low-key as strep throat and a patient may want to protect certain topics from being discussed over a “non-secure” app but why not let the patient decide through informed consent?.

Regulators could also abandon this all-or-nothing approach and lighten regulations surrounding specific health conditions. The idea low cost vibramycin that regulations change based on medical situation is not new. For example, in my home state of Michigan, adolescents are essentially considered emancipated if it involves sexual health, mental health or substance abuse. Never mind that this same information is freely given over the phone by every office around the country daily without issue, but I digress. While my job is to innovate new pathways for care, our lawyer’s job is to protect the low cost vibramycin organization and he, along with IT security, rightfully shot down my consumer applications idea.

A few days later I legitimately screamed out loud in joy when the Department of Health and Human Services announced that it would use discretion on enforcing HIPAA compliance rules and specifically allowed for use of consumer applications. The elimination of billing restrictions and HIPAA regulations changed what is possible for health care organizations to offer virtually. Unfortunately both changes are listed as temporary and low cost vibramycin will likely be removed when the pandemic ends. Six days after the HIPAA changes were announced, we launched a centralized virtual clinic for any patient that wanted a direct-to-consumer video visit to be screened by a provider for COVID-19. It allows patients to call in without a referral and most patients are on-screen within five minutes of clicking the link we text them.

They don’t low cost vibramycin have to download an app, create an account or even be an established patient of our health system. It saw over 900 patients in the first 12 days it was open. That is 900 real patients that received care from a physician or advanced practice provider without risking personal exposure and without going to an already overwhelmed ED or urgent care. To date, 70 percent of the patients seen by the virtual clinic did not meet CDC testing criteria low cost vibramycin for COVID-19. I don’t believe we could have reached even half of these patients had the consumer application restrictions been kept.

A program like this almost certainly wouldn’t exist if not for the regulations being lifted and even if it did, it would have taken six to 12 months to navigate barriers and implement in normal times. Sure, the low cost vibramycin urgency of a pandemic helps but the impact of provider, patients, regulators and payors being on the same page is what fueled this fire. During the virtual clinic’s first two weeks, my team turned its attention to getting over 300 providers across 60+ offices virtual so they could see their patients at home. Imagine being an immunocompromised cancer patient right now and being asked to leave your home and be exposed to other people in order to see your oncologist. Direct-to-consumer virtual care is the best way to safely care for these patients and low cost vibramycin without these temporary waivers it wouldn’t be covered by insurance even if you did navigate the clunky apps that are HIPAA compliant.

Do we really think the immunocompromised cancer patient feels any more comfortable every normal flu season?. Is it any more appropriate to ask them to risk exposure to the flu than it is to COVID-19?. And yet we deny them this access in normal times and it quite possibly will be stripped away from them when this low cost vibramycin crisis is over. Now 300 to 400 patients per day in our health system are seen virtually by their own primary care doctor or specialist for non-COVID related visits. Not a single one of these would have been reimbursed one month ago and I am highly skeptical I would have gotten approval to use the software that connects us to the patient.

Lastly, recall that prior low cost vibramycin to COVID-19, our system had only found 250 total patients that direct-to-consumer care was value-added and wasn’t restricted by regulation or reimbursement. COVID-19 has been a wake-up call to the whole country and health care is no exception. It has put priorities in perspective and shined a light on what is truly value-added. For direct-to-consumer low cost vibramycin virtual care it has shown us what is possible when we get out of our own way. If a regulation has to be removed to allow for care during a crisis then we must question why it exists in the first place.

HIPAA regulation cannot go back to its antiquated practices if we are truly going to shift the focus to patient wellness. CMS and private payors must embrace value-added direct-to-consumer virtual care and allow low cost vibramycin patients the access they deserve. COVID-19 has forced this industry forward, we cannot allow it to regress and be forgotten when this is over. Tom Wood is the director of trauma and virtual care for MidMichigan Health, a non-profit health system headquartered in Midland, Michigan, affiliated with Michigan Medicine, the health care division of the University of Michigan. The views and low cost vibramycin opinions expressed in this commentary are his own.When dealing with all of the aspects of diabetes, it’s easy to let your feel fall to the bottom of the list.

But daily care and evaluation is one of the best ways to prevent foot complications. It’s important to identify your risk factors and take the proper steps in limiting your complications. Two of the low cost vibramycin biggest complications with diabetes are peripheral neuropathy and ulcer/amputation. Symptoms of peripheral neuropathy include numbness, tingling and/or burning in your feet and legs. You can slow the progression of developing neuropathy by making it a point to manage your blood sugars and keep them in the normal range.

If you are experiencing these low cost vibramycin symptoms, it is important to establish and maintain a relationship with a podiatrist. Your podiatrist can make sure things are looking healthy and bring things to your attention to monitor and keep a close eye on. Open wounds or ulcers can develop secondary to trauma, pressure, diabetes, neuropathy or poor circulation. If ulcerations do develop, it’s extremely important to identify the cause and low cost vibramycin address it. Ulcers can get worse quickly, so it’s necessary to seek immediate medical treatment if you find yourself or a loved one dealing with this complication.

Untreated ulcerations often lead to amputation and can be avoided if proper medical attention is sought right away. There are low cost vibramycin important things to remember when dealing with diabetic foot care. It’s very important to inspect your feet daily, especially if you have peripheral neuropathy. You may have a cut or a sore on your feet that you can’t feel, so your body doesn’t alarm you to check your feet. Be gentle when low cost vibramycin bathing your feet.

Moisturize your feet, but not between your toes. Do not treat calluses or corns on your own. Wear clean, low cost vibramycin dry socks. Never walk barefoot, and consider socks and shoes made specifically for patients with diabetes. Kristin Raleigh, D.P.M., is a podiatrist who sees patients at Foot &.

Ankle Specialists of Mid-Michigan in Midland.

Vibramycin d

Air conditioning and other vibramycin d cooling systems are widely recognized as integral to protecting people from the sometimes deadly impacts of extreme heat, which are intensifying in step with climate change. Yet according to a study, published yesterday in Nature Sustainability, there remains a “global blind spot” when it comes to handling the already exorbitant demand for cooling and indoor air conditioning, which alone is projected to triple by 2050. That’s a stark reality, the report warns, given that many vibramycin d cooling systems are carbon-intensive—and contribute to global warming themselves. €œCooling is essential to human well-being and health, from the food we eat to the storage of medicine to how comfortable and productive we are at home, school or the office,” said report co-author Radhika Khosla, a principal investigator at the Oxford Martin Programme on the Future of Cooling.

So if societies do not soon begin implementing sustainable cooling solutions, Khosla added in a statement, they risk “locking the world into a deadly feedback loop, where demand for cooling energy drives further greenhouse gas emissions and results in even more global warming.” The researchers examined thousands of peer-reviewed papers related to the United Nations’ Sustainable Development Goals and concluded that greener cooling systems could help achieve all 17 goals—which include curbing global hunger, reducing gender inequality and improving human health writ large. That’s possible, the report said, because extreme heat dramatically affects vibramycin d everything from food production to water quality to students’ ability to learn and focus during school. Despite evidence that demonstrates the connection between efficient cooling systems and improved social and environmental outcomes, however, the authors argue that the “unprecedented rise in demand and the potential benefits of sustainable cooling” remain largely neglected in contemporary sustainability debates. That has major implications, vibramycin d they emphasized, for sustainable development around the world.

To close that gap, the study said technological developments, innovative business models, intentional infrastructure and regulation could be used to make cooling more accessible—and climate friendly. Cities and towns, for instance, could embed “passive and energy-efficient” cooling mechanisms in urban infrastructure to lessen the impact of extreme heat both indoors and outdoors. That could entail projects intended to reduce the prominence of “urban heat vibramycin d islands” by planting additional trees, developing new parks and building green roofs—all of which naturally cool urban spaces. Those strategies would be especially useful, the report said, given that “projections of the world’s population living in towns and cities are set to reach 66% by 2050,” making urban areas the “epicentre of cooling demand.” The authors also suggest that air-conditioning companies adopt a “cooling as a service” business model, intended to making sustainable cooling more affordable—especially in hot, low-income regions.

Rather than charging for the system itself, the companies would profit by retaining ownership of it and charging customers to operate the system and maintain a comfortable thermal environment. This would vibramycin d drive down, or even eliminate, what can be prohibitive upfront costs for cash-strapped households. In the context of a world “positioned at the brink of unprecedented cooling demand,” the report says, these interventions are among the many that offer “a way forward while being acutely aware of the extraordinary opportunity the current moment provides to use cooling as a lens to look to the sustainability of our future.” Reprinted from Climatewire with permission from E&E News. E&E provides daily coverage vibramycin d of essential energy and environmental news at www.eenews.net.Tempers are running hot in science (as they are in the U.S.

At large) as the field embarks on a long-overdue conversation about its treatment of women and people of color. In June, for example, thousands of researchers and academics across the globe—as well as the preeminent journals Science and Nature—stopped work for a day to protest racism in their ranks. The American Physical Society endorsed the effort to “shut down STEM,” declaring its commitment to “eradicating systemic racism and discrimination” in science vibramycin d. Physics exemplifies the problem.

African-Americans make up about 14 percent of the college-age population in the U.S., commensurate with their numbers in the overall population, but in physics they receive 3 to 4 percent of undergraduate degrees and less than 3 percent of Ph.D.s, and as of 2012 they composed only 2 percent of faculty. No doubt there are many reasons for this underrepresentation, but one troubling factor is vibramycin d the refusal of some scientists to acknowledge that a problem could even exist. Science, they argue, is inherently rational and self-correcting. Would that vibramycin d were true.

The history of science is rife with well-documented cases of misogyny, prejudice and bias. For centuries biologists promoted false theories of female inferiority, and scientific institutions typically barred women's participation. Historian of science and MacArthur fellow Margaret Rossiter has documented how, in the mid-19th vibramycin d century, female scientists created their own scientific societies to compensate for their male colleagues' refusal to acknowledge their work. Sharon Bertsch McGrayne filled an entire volume with the stories of women who should have been awarded the Nobel Prize for work that they did in collaboration with male colleagues—or, worse, that they had stolen by them.

(Rosalind Franklin is a well-documented example of the latter. Her photographs of the crystal structure of DNA were vibramycin d shared without her permission by one of the men who then won the Nobel Prize for elucidating the double-helix structure.) Racial bias has been at least as pernicious as gender bias. It was scientists, after all, who codified the concept of race as a biological category that was not simply descriptive but also hierarchical. Good scientists are open to competing ideas vibramycin d.

They attend to challenging data, and they listen to opposing views. But scientists are also humans, and cognitive science shows that humans are prone to bias, misperception, motivated reasoning and other intellectual pitfalls. Because reasoning is slow and difficult, we rely on heuristics—intellectual shortcuts that often vibramycin d work but sometimes fail spectacularly. (Believing that men are, in general, better than women in math is one tiring example.) It is not credible to claim that scientists are somehow immune to the biases that afflict everyone else.

Fortunately, the objectivity of scientific knowledge does not depend vibramycin d on the objectivity of individual scientists. Rather it depends on strategies for identifying, acknowledging and correcting bias and error. As I point out in my 2019 book, Why Trust Science, scientific knowledge begins as claims advanced by individual scientists, teams or laboratories that are then closely scrutinized by others, who may bring forward additional proof to sustain them—or to modify or reject them. What emerges vibramycin d as a scientific fact or established theory is rarely if ever the same as the starting claim.

It has been adjusted in light of evidence and argumentation. Science is a collective effort, and it works best when scientific communities are diverse. The reason is simple vibramycin d. Heterogeneous communities are more likely than homogeneous ones to be able to identify blind spots and correct them.

Science does not correct itself vibramycin d. Scientists correct one another through critical interrogation. And that means being willing to interrogate not just claims about the external world but claims about our own practices and processes as well. Science has an admirable record of producing reliable knowledge about the natural and social vibramycin d world, but not when it comes to acknowledging its own weaknesses.

And we cannot correct those weaknesses if we insist the system will magically correct itself. It is not ideological to acknowledge and confront bias in science. It is ideological to insist science cannot be biased vibramycin d despite empirical validation to the contrary. Given that our failings of inclusion have been known for a long time, it is high time we finally fix them.Young, healthy people will be intentionally exposed to the virus responsible for COVID-19 in a first-of-its kind ‘human challenge trial’, the UK government and a company that runs such studies announced on 20 October.

The experiment, vibramycin d set to begin in January in a London hospital if it receives final regulatory and ethical approval, aims to accelerate the development of vaccines that could end the pandemic. Human challenge trials have a history of providing insight into diseases such as malaria and influenza. The UK trial will try to identify a suitable dose of the virus SARS-CoV-2 that could be used in future vaccine trials. But the prospect of deliberately infecting people—even those at low risk of severe disease—with SARS-CoV-2, a deadly pathogen that has few proven treatments, is uncharted vibramycin d medical and bioethical territory.

Proponents of COVID-19 challenge trials have argued that they can be run safely and ethically, and that their potential to quickly identify effective vaccines outweighs the low risks to participants. But others have raised questions about the safety and value of these studies, pointing out that large-scale efficacy trials involving tens of thousands of people are expected to deliver results on several COVID-19 vaccines soon. €œDeliberately infecting volunteers with a known human pathogen is never undertaken lightly vibramycin d. However, such studies are enormously informative about a disease,” said Peter Openshaw, an immunologist at Imperial College London and investigator on the study, in a press statement.

€œIt is really vital that we move as fast as possible towards getting effective vibramycin d vaccines and other treatments for COVID-19, and challenge studies have the potential to accelerate and de-risk the development of novel drugs and vaccines.” Dose testing The planned COVID-19 challenge study will be led by a Dublin-based commercial clinical-research organization called Open Orphan and its subsidiary hVIVO, which runs challenge trials on respiratory pathogens. It will take place in the high-level isolation unit of the Royal Free Hospital in north London, says Open Orphan executive chair Cathal Friel. The UK government’s COVID-19 Vaccine Taskforce has agreed to pay the company up to £10 million (US$13 million) to conduct the trial, with the possibility of contracting Open Orphan to run several more to test various vaccines. The UK Medicines and Healthcare Regulatory Agency (MHRA), which regulates clinical trials in the United Kingdom, and an ethical review committee, still need to approve the initial trial and its vibramycin d design, and that of future studies.

The initial trial will involve an estimated 30–50 participants, says Andrew Catchpole, a virologist and the chief scientific officer at Open Orphan who is leading the work. It is open only to vibramycin d healthy adults aged 18–30. The precise design of the study has not been finalized. But it is likely that a small number of participants will receive a very low dose of a SARS-CoV-2 ‘challenge strain’ derived from a currently circulating virus and grown under stringent conditions.

If none or few of the participants become infected, the vibramycin d researchers will seek permission from an independent safety monitoring board to expose participants to higher doses. This process will be repeated until researchers identify a dose that infects most of those exposed, says Catchpole. Once an appropriate dose is identified, Open Orphan could be asked to run a series of challenge trials testing several vaccines. Catchpole says that the design of vibramycin d these trials, including which vaccines will be included, has not been determined.

He envisions that some trial participants will receive a placebo injection instead of a vaccine, but he also says that head-to-head trials comparing two or more vaccines could be run. Other vaccine studies that the company runs typically enrol 40–50 volunteers for vibramycin d each trial arm, he says. Catchpole says that his team will take every precaution against participants in the initial trial developing severe disease. Volunteers will be treated with an antiviral drug, such as remdesivir, once a nasal swab gives a positive result for SARS-CoV-2 genetic material.

In addition to age and health, participants will be screened for risk factors that have been vibramycin d associated with severe COVID-19. Selecting participants at the lowest risk is the most important safety step in running a challenge trial, says Matt Memoli, an infectious-disease physician and virologist at the US National Institute of Allergy and Infectious Diseases (NIAID) in Bethesda, Maryland. €œOnce you’ve given that virus to the person, anything’s possible,” he says. €œYou can’t control it, you can only react to it.” If Open Orphan moves on to vaccine trials, it will vibramycin d aim to recruit around 500 participants altogether, but Friel says the company will need to screen many times more people to identify suitable volunteers.

An ethical review board will determine how to compensate participants. Open Orphan vibramycin d typically pays volunteers around £4,000 for their time, says Catchpole. Ethical issues There is a concern that people will participate for the money without appreciating the risks, says Nir Eyal, a bioethicist at Rutgers University in New Brunswick, New Jersey, who has argued that COVID-19 challenge trials can be run safely and ethically. But a well-designed online course, for instance, could ensure that participants understand the risks, he says.

Ensuring that vibramycin d participants understand the limitations of challenge trials will also be important, says Seema Shah, a bioethicist at Lurie Children’s Hospital and Northwestern University in Chicago, Illinois. With phase III trials of numerous COVID-19 vaccines in the works, she thinks it unlikely that challenge trials will speed the development of the first vaccines. Instead, their payoff could lie in helping to test later-generation vaccines or laying the groundwork for fresh insights into the disease. In this context, says vibramycin d Shah, “It becomes a little bit harder to justify them, and we need to take a close look at risks.” Meagan Deming, a vaccine scientist and virologist at the University of Maryland School of Medicine in Baltimore, sees challenge trials as more appropriate for studying basic aspects of SARS-CoV-2 infection—such as the potential for reinfection or how previous exposure to cold-causing coronaviruses influences susceptibility to COVID-19—than for vetting vaccines.

Because such trials are likely to involve only young, healthy people, they might not reveal much about how vaccines could protect those most at risk of severe disease, such as older people and those with conditions such as diabetes, Deming says. €œThere’s a reason we don’t have a lot of vaccines approved by challenge models, because they vibramycin d don’t apply to everyone and you want a vaccine to protect almost everyone,” she says. Phase III trials might not offer clear evidence of whether vaccines work in older people, because of their low participation in those trials, says Peter Smith, an epidemiologist at the London School of Hygiene and Tropical Medicine who has been involved in challenge trials. Researchers will probably need to determine vaccines’ likely effectiveness in older people, on the basis of how their immune systems respond to COVID-19 vaccines before exposure to the virus.

And compared with field trials, challenge studies are better at identifying the types of immune response that predict whether a vaccine is likely to work or vibramycin d not, adds Memoli. Other trials The United Kingdom isn’t the only country investigating COVID-19 challenge trials. Belgium’s government has committed €20 million (US$23.6 million) for facilities to host challenge trials, potentially involving COVID-19. NIAID is funding the development of two SARS-CoV-2 challenge strains by a lab at vibramycin d Colorado State University in Fort Collins, and a team led by Memoli is also laying the groundwork for such trials.

In a statement, NIAID said it was awaiting data from phase III studies before making decisions on COVID-19 challenge trials. Proponents of the trials argue that the consequences of delaying them should be taken into account, alongside vibramycin d the risks of going forward. For instance, Eyal and economists Pedro Rosa Dias and Ara Darzi at Imperial College London have calculated that speeding up the development of COVID-19 vaccines by one month would avert the loss of 720,000 years of life and prevent 40 million years in poverty, mostly in lower-income countries. But Deming thinks that challenge trials should wait until their value is clearer and the risks can be better mitigated, for instance by deploying more potent therapies.

€œWe don’t yet know enough about this disease vibramycin d to say for this person. You will not die,” she says. €œWe’ve learned so much in the past nine months vibramycin d. In a year, we will be able to do this safely.” This article is reproduced with permission and was first published on October 20 2020.For the first time ever, a NASA probe has performed a sample-snagging operation on an asteroid in deep space.

The agency’s OSIRIS-REx spacecraft spiraled down to the surface of the near-Earth asteroid Bennu this afternoon (Oct. 20) to grab material vibramycin d that mission team members hope harbors clues about the solar system’s early days and the rise of life on Earth. €œWe did it!. € OSIRIS-REx principal investigator Dante Lauretta, of the University of Arizona, said during a webcast that provided updates about today’s maneuver.

€œWe tagged the surface of the asteroid, and it’s up to Bennu now to see how the event went.” vibramycin d The goal was to collect at least 60 grams (2.1 ounces) of dirt and gravel from Bennu’s rubbly surface. It could take up to 10 days to determine if OSIRIS-REx achieved this aim, mission team members have said. And it’s vibramycin d not a disaster if the asteroid haul turns out to be a little light. The probe can go back down for two more tries if need be.

“This amazing first for NASA demonstrates how an incredible team from across the country came together and persevered through incredible challenges to expand the boundaries of knowledge,” NASA Administrator Jim Bridenstine said in a statement after the touchdown. €œOur industry, vibramycin d academic, and international partners have made it possible to hold a piece of the most ancient solar system in our hands.” Lauretta and his fellow OSIRIS-REx scientists and engineers watched over today’s asteroid sample-snatching attempt from a mission operations center at Lockheed Martin Space in Littleton, Colorado. (Lockheed Martin built the spacecraft for NASA.) And while the mood was certainly jubilant, the impact of the ongoing COVID-19 pandemic was clear. For example, everyone wore facemasks and maintained appropriate social distancing for much of the event.

While there were some hugs after news of OSIRIS-REx’s asteroid touchdown, they were few in NASA’s live webcast, with hand sanitizer clearly vibramycin d on hand after such celebrations. “This is one of those moments where we’re all aware of COVID-19,” NASA astronomer Michelle Thaller of the Goddard Space Flight Center said in the webcast just after touchdown. €œBecause I want the hugs and the high fives and everything, but we’re all going to keep each other safe.” The $800 million OSIRIS-REx vibramycin d mission launched in September 2016 and arrived at the 1,640-foot-wide (500 meters) Bennu in December 2018. The probe has been taking the asteroid’s measure ever since, mapping its surface in incredible detail to prepare for today’s maneuver.

That work has revealed a world far more rugged than the mission team had expected. House-sized boulders stud Bennu’s surface, limiting the available options for a safe sample vibramycin d grab. The team eventually homed in on a small crater called Nightingale as its top choice, because the site sports relatively fresh and fine-grained material that hasn’t been exposed to the harsh deep-space environment for long. But Nightingale is surrounded by hazards, including a big outcrop the mission team nicknamed “Mount Doom.” There are obstacles within the crater as well, so the spacecraft targeted a relatively flat, boulder-free area just 26 feet (8 m) wide—quite an ambitious goal, considering that OSIRIS-REx is the size of a 15-passenger van and the original mission plan envisioned a touchdown zone 165 feet (50 m) wide.

€œSo, for vibramycin d some perspective. The next time you park your car in front of your house or in front of a coffee shop and walk inside, think about the challenge of navigating OSIRIS-REx into one of these spots from 200 million miles away,” Mike Moreau, OSIRIS-REx deputy project manager at NASA’s Goddard Space Flight Center in Greenbelt, Maryland, said during a news conference last month. It currently takes more than 18 minutes for commands to travel from Earth to vibramycin d OSIRIS-REx, so Moreau and his colleagues cannot control the probe in real time. The craft therefore performed today’s operation autonomously.

Shortly before 2 p.m. EDT (1600 GMT) today, OSIRIS-REx fired its thrusters to get out vibramycin d of orbit around Bennu and head down toward the surface. At 6:12 p.m. EDT (2212 GMT), the probe “kissed” the asteroid for about 10 seconds with its sample-collecting mechanism, which is affixed to the end of OSIRIS-REx’s 11-foot-long (3.4 vibramycin d m) robotic arm.

During the brief touchdown, the spacecraft blasted Bennu’s surface with nitrogen gas. This stirred up dirt and rock that could then be collected by the arm’s sampling head, which mission team members have likened to an older car’s air filter. We should expect OSIRIS-REx’s first images of the operation to vibramycin d start coming down to Earth tomorrow morning (Oct. 21), mission team members said.

The OSIRIS-REx team will spend the next week or so assessing how much asteroid material was collected. The probe’s handlers have expressed vibramycin d confidence that this first attempt will succeed. OSIRIS-REx’s sampler was designed to snag at least 150 grams (5.3 ounces) and could theoretically get up to 4 kilograms (8.8 lbs.) of material if everything went perfectly. But if OSIRIS-REx is deemed to have come up short on collected material today, another attempt could be made, at a backup site known as Osprey, as vibramycin d soon as January 2021.

A third try would be possible, too, if needed. The probe carries three bottles of surface-disturbing nitrogen gas. Those are contingency plans, vibramycin d however. If things went according to plan today, OSIRIS-REx remains on course to depart Bennu in March 2021.

The collected samples are scheduled to land here on Earth, encased in a special return capsule, in September 2023. Scientists will then study the material vibramycin d in labs around the world, scrutinizing the stuff in far more detail than OSIRIS-REx, or any other single probe, could do on its own in deep space. Asteroids are building blocks left over from the planet-formation epoch, so such analyses could reveal key insights about our solar system’s very early days, NASA officials have said. €œThis was an incredible feat—and today we’ve advanced both science and engineering and vibramycin d our prospects for future missions to study these mysterious ancient storytellers of the solar system,” said Thomas Zurbuchen, NASA’s associate administrator for science missions, in the NASA statement.

€œA piece of primordial rock that has witnessed our solar system’s entire history may now be ready to come home for generations of scientific discovery, and we can’t wait to see what comes next.” In addition, Bennu is rich in hydrated minerals and carbon-containing organic compounds. Asteroids like it may have helped Earth become habitable long ago, seeding our planet with the ingredients needed for life as we know it. €œAnd also, having the samples back vibramycin d here on Earth allows us to preserve them for future generations to come and allows for future explorers to analyze the samples using techniques and instruments that haven’t been invented, and to ask questions that we don’t even know to ask yet,” Lori Glaze, director of NASA’s Planetary Science Division, said during today’s webcast. Getting these samples down to Earth is OSIRIS-REx’s top priority.

But the mission also has other goals, as indicated by its full name—“Origins, Spectral Interpretation, Resource Identification, Security, Regolith Explorer.” For example, observations the probe has made while orbiting Bennu should help scientists better understand how asteroids move through space, NASA officials have said. This information could improve trajectory projections for potentially vibramycin d hazardous asteroids, a category that includes Bennu. (There’s a 1-in-2,700 chance that Bennu will hit Earth during a close approach in the late 2100s, researchers say.) OSIRIS-REx’s sample won’t be the first pristine asteroid material brought down to Earth by a space mission. Japan’s Hayabusa probe returned some grains of the stony asteroid Itokawa in 2010, and its successor, Hayabusa2, recently grabbed pieces of the carbon-rich rock Ryugu vibramycin d.

The material from Ryugu is scheduled to land on Earth this December. The OSIRIS-REx and Hayabusa2 teams have been working together for the past few years, and that collaboration will continue after the missions’ samples touch down on Earth, NASA officials have stressed. Copyright 2020 Space.com, a Future company vibramycin d. All rights reserved.

This material may not be published, broadcast, rewritten or redistributed..

Air conditioning and other cooling systems are widely recognized as integral to protecting people from the sometimes deadly impacts of extreme low cost vibramycin heat, which are intensifying in step with climate change. Yet according to a study, published yesterday in Nature Sustainability, there remains a “global blind spot” when it comes to handling the already exorbitant demand for cooling and indoor air conditioning, which alone is projected to triple by 2050. That’s a low cost vibramycin stark reality, the report warns, given that many cooling systems are carbon-intensive—and contribute to global warming themselves.

€œCooling is essential to human well-being and health, from the food we eat to the storage of medicine to how comfortable and productive we are at home, school or the office,” said report co-author Radhika Khosla, a principal investigator at the Oxford Martin Programme on the Future of Cooling. So if societies do not soon begin implementing sustainable cooling solutions, Khosla added in a statement, they risk “locking the world into a deadly feedback loop, where demand for cooling energy drives further greenhouse gas emissions and results in even more global warming.” The researchers examined thousands of peer-reviewed papers related to the United Nations’ Sustainable Development Goals and concluded that greener cooling systems could help achieve all 17 goals—which include curbing global hunger, reducing gender inequality and improving human health writ large. That’s possible, the report said, because extreme heat dramatically affects everything from food production to low cost vibramycin water quality to students’ ability to learn and focus during school.

Despite evidence that demonstrates the connection between efficient cooling systems and improved social and environmental outcomes, however, the authors argue that the “unprecedented rise in demand and the potential benefits of sustainable cooling” remain largely neglected in contemporary sustainability debates. That has major implications, they emphasized, for sustainable development around the world low cost vibramycin. To close that gap, the study said technological developments, innovative business models, intentional infrastructure and regulation could be used to make cooling more accessible—and climate friendly.

Cities and towns, for instance, could embed “passive and energy-efficient” cooling mechanisms in urban infrastructure to lessen the impact of extreme heat both indoors and outdoors. That could entail projects intended to reduce the prominence of low cost vibramycin “urban heat islands” by planting additional trees, developing new parks and building green roofs—all of which naturally cool urban spaces. Those strategies would be especially useful, the report said, given that “projections of the world’s population living in towns and cities are set to reach 66% by 2050,” making urban areas the “epicentre of cooling demand.” The authors also suggest that air-conditioning companies adopt a “cooling as a service” business model, intended to making sustainable cooling more affordable—especially in hot, low-income regions.

Rather than charging for the system itself, the companies would profit by retaining ownership of it and charging customers to operate the system and maintain a comfortable thermal environment. This would drive down, or even eliminate, what low cost vibramycin can be prohibitive upfront costs for cash-strapped households. In the context of a world “positioned at the brink of unprecedented cooling demand,” the report says, these interventions are among the many that offer “a way forward while being acutely aware of the extraordinary opportunity the current moment provides to use cooling as a lens to look to the sustainability of our future.” Reprinted from Climatewire with permission from E&E News.

E&E provides daily coverage of essential energy and environmental news at www.eenews.net.Tempers are running hot in science (as they are in the U.S low cost vibramycin. At large) as the field embarks on a long-overdue conversation about its treatment of women and people of color. In June, for example, thousands of researchers and academics across the globe—as well as the preeminent journals Science and Nature—stopped work for a day to protest racism in their ranks.

The American low cost vibramycin Physical Society endorsed the effort to “shut down STEM,” declaring its commitment to “eradicating systemic racism and discrimination” in science. Physics exemplifies the problem. African-Americans make up about 14 percent of the college-age population in the U.S., commensurate with their numbers in the overall population, but in physics they receive 3 to 4 percent of undergraduate degrees and less than 3 percent of Ph.D.s, and as of 2012 they composed only 2 percent of faculty.

No doubt there are low cost vibramycin many reasons for this underrepresentation, but one troubling factor is the refusal of some scientists to acknowledge that a problem could even exist. Science, they argue, is inherently rational and self-correcting. Would that were low cost vibramycin true.

The history of science is rife with well-documented cases of misogyny, prejudice and bias. For centuries biologists promoted false theories of female inferiority, and scientific institutions typically barred women's participation. Historian of science and low cost vibramycin MacArthur fellow Margaret Rossiter has documented how, in the mid-19th century, female scientists created their own scientific societies to compensate for their male colleagues' refusal to acknowledge their work.

Sharon Bertsch McGrayne filled an entire volume with the stories of women who should have been awarded the Nobel Prize for work that they did in collaboration with male colleagues—or, worse, that they had stolen by them. (Rosalind Franklin is a well-documented example of the latter. Her photographs of the crystal structure of DNA were shared without her permission by one of the men who then won the Nobel Prize for elucidating the double-helix structure.) Racial bias has been at least low cost vibramycin as pernicious as gender bias.

It was scientists, after all, who codified the concept of race as a biological category that was not simply descriptive but also hierarchical. Good scientists are open to competing low cost vibramycin ideas. They attend to challenging data, and they listen to opposing views.

But scientists are also humans, and cognitive science shows that humans are prone to bias, misperception, motivated reasoning and other intellectual pitfalls. Because reasoning is slow and difficult, we rely low cost vibramycin on heuristics—intellectual shortcuts that often work but sometimes fail spectacularly. (Believing that men are, in general, better than women in math is one tiring example.) It is not credible to claim that scientists are somehow immune to the biases that afflict everyone else.

Fortunately, the objectivity low cost vibramycin of scientific knowledge does not depend on the objectivity of individual scientists. Rather it depends on strategies for identifying, acknowledging and correcting bias and error. As I point out in my 2019 book, Why Trust Science, scientific knowledge begins as claims advanced by individual scientists, teams or laboratories that are then closely scrutinized by others, who may bring forward additional proof to sustain them—or to modify or reject them.

What emerges as a scientific fact or established theory is rarely low cost vibramycin if ever the same as the starting claim. It has been adjusted in light of evidence and argumentation. Science is a collective effort, and it works best when scientific communities are diverse.

The reason low cost vibramycin is simple. Heterogeneous communities are more likely than homogeneous ones to be able to identify blind spots and correct them. Science does not correct low cost vibramycin itself.

Scientists correct one another through critical interrogation. And that means being willing to interrogate not just claims about the external world but claims about our own practices and processes as well. Science has an admirable record of producing reliable knowledge about the natural and low cost vibramycin social world, but not when it comes to acknowledging its own weaknesses.

And we cannot correct those weaknesses if we insist the system will magically correct itself. It is not ideological to acknowledge and confront bias in science. It is ideological to insist low cost vibramycin science cannot be biased despite empirical validation to the contrary.

Given that our failings of inclusion have been known for a long time, it is high time we finally fix them.Young, healthy people will be intentionally exposed to the virus responsible for COVID-19 in a first-of-its kind ‘human challenge trial’, the UK government and a company that runs such studies announced on 20 October. The experiment, set to begin in January in a London hospital if it receives final regulatory and ethical approval, aims to accelerate the development of low cost vibramycin vaccines that could end the pandemic. Human challenge trials have a history of providing insight into diseases such as malaria and influenza.

The UK trial will try to identify a suitable dose of the virus SARS-CoV-2 that could be used in future vaccine trials. But the prospect of deliberately infecting people—even those at low risk of severe disease—with SARS-CoV-2, a deadly pathogen that has few proven treatments, is low cost vibramycin uncharted medical and bioethical territory. Proponents of COVID-19 challenge trials have argued that they can be run safely and ethically, and that their potential to quickly identify effective vaccines outweighs the low risks to participants.

But others have raised questions about the safety and value of these studies, pointing out that large-scale efficacy trials involving tens of thousands of people are expected to deliver results on several COVID-19 vaccines soon. €œDeliberately infecting low cost vibramycin volunteers with a known human pathogen is never undertaken lightly. However, such studies are enormously informative about a disease,” said Peter Openshaw, an immunologist at Imperial College London and investigator on the study, in a press statement.

€œIt is really vital that we move as fast as possible towards getting effective vaccines and other treatments for COVID-19, and challenge studies have the potential to accelerate and de-risk the development of novel drugs and vaccines.” Dose testing The planned low cost vibramycin COVID-19 challenge study will be led by a Dublin-based commercial clinical-research organization called Open Orphan and its subsidiary hVIVO, which runs challenge trials on respiratory pathogens. It will take place in the high-level isolation unit of the Royal Free Hospital in north London, says Open Orphan executive chair Cathal Friel. The UK government’s COVID-19 Vaccine Taskforce has agreed to pay the company up to £10 million (US$13 million) to conduct the trial, with the possibility of contracting Open Orphan to run several more to test various vaccines.

The UK Medicines and Healthcare Regulatory Agency (MHRA), which regulates low cost vibramycin clinical trials in the United Kingdom, and an ethical review committee, still need to approve the initial trial and its design, and that of future studies. The initial trial will involve an estimated 30–50 participants, says Andrew Catchpole, a virologist and the chief scientific officer at Open Orphan who is leading the work. It is low cost vibramycin open only to healthy adults aged 18–30.

The precise design of the study has not been finalized. But it is likely that a small number of participants will receive a very low dose of a SARS-CoV-2 ‘challenge strain’ derived from a currently circulating virus and grown under stringent conditions. If none or few of the participants become infected, the researchers will seek permission low cost vibramycin from an independent safety monitoring board to expose participants to higher doses.

This process will be repeated until researchers identify a dose that infects most of those exposed, says Catchpole. Once an appropriate dose is identified, Open Orphan could be asked to run a series of challenge trials testing several vaccines. Catchpole says that the design low cost vibramycin of these trials, including which vaccines will be included, has not been determined.

He envisions that some trial participants will receive a placebo injection instead of a vaccine, but he also says that head-to-head trials comparing two or more vaccines could be run. Other vaccine studies low cost vibramycin that the company runs typically enrol 40–50 volunteers for each trial arm, he says. Catchpole says that his team will take every precaution against participants in the initial trial developing severe disease.

Volunteers will be treated with an antiviral drug, such as remdesivir, once a nasal swab gives a positive result for SARS-CoV-2 genetic material. In addition to age and health, participants low cost vibramycin will be screened for risk factors that have been associated with severe COVID-19. Selecting participants at the lowest risk is the most important safety step in running a challenge trial, says Matt Memoli, an infectious-disease physician and virologist at the US National Institute of Allergy and Infectious Diseases (NIAID) in Bethesda, Maryland.

€œOnce you’ve given that virus to the person, anything’s possible,” he says. €œYou can’t control it, you can only low cost vibramycin react to it.” If Open Orphan moves on to vaccine trials, it will aim to recruit around 500 participants altogether, but Friel says the company will need to screen many times more people to identify suitable volunteers. An ethical review board will determine how to compensate participants.

Open Orphan typically low cost vibramycin pays volunteers around £4,000 for their time, says Catchpole. Ethical issues There is a concern that people will participate for the money without appreciating the risks, says Nir Eyal, a bioethicist at Rutgers University in New Brunswick, New Jersey, who has argued that COVID-19 challenge trials can be run safely and ethically. But a well-designed online course, for instance, could ensure that participants understand the risks, he says.

Ensuring that participants understand the limitations of challenge trials will also be important, says Seema Shah, a bioethicist at Lurie Children’s Hospital and Northwestern University in Chicago, Illinois low cost vibramycin. With phase III trials of numerous COVID-19 vaccines in the works, she thinks it unlikely that challenge trials will speed the development of the first vaccines. Instead, their payoff could lie in helping to test later-generation vaccines or laying the groundwork for fresh insights into the disease.

In this context, says Shah, “It low cost vibramycin becomes a little bit harder to justify them, and we need to take a close look at risks.” Meagan Deming, a vaccine scientist and virologist at the University of Maryland School of Medicine in Baltimore, sees challenge trials as more appropriate for studying basic aspects of SARS-CoV-2 infection—such as the potential for reinfection or how previous exposure to cold-causing coronaviruses influences susceptibility to COVID-19—than for vetting vaccines. Because such trials are likely to involve only young, healthy people, they might not reveal much about how vaccines could protect those most at risk of severe disease, such as older people and those with conditions such as diabetes, Deming says. €œThere’s a reason we don’t have a lot of vaccines approved by challenge models, because they don’t apply to everyone and you want a vaccine to protect almost everyone,” she says low cost vibramycin.

Phase III trials might not offer clear evidence of whether vaccines work in older people, because of their low participation in those trials, says Peter Smith, an epidemiologist at the London School of Hygiene and Tropical Medicine who has been involved in challenge trials. Researchers will probably need to determine vaccines’ likely effectiveness in older people, on the basis of how their immune systems respond to COVID-19 vaccines before exposure to the virus. And compared with field trials, challenge studies are better at identifying the types of immune low cost vibramycin response that predict whether a vaccine is likely to work or not, adds Memoli.

Other trials The United Kingdom isn’t the only country investigating COVID-19 challenge trials. Belgium’s government has committed €20 million (US$23.6 million) for facilities to host challenge trials, potentially involving COVID-19. NIAID is funding the development of two SARS-CoV-2 challenge strains by a lab low cost vibramycin at Colorado State University in Fort Collins, and a team led by Memoli is also laying the groundwork for such trials.

In a statement, NIAID said it was awaiting data from phase III studies before making decisions on COVID-19 challenge trials. Proponents of low cost vibramycin the trials argue that the consequences of delaying them should be taken into account, alongside the risks of going forward. For instance, Eyal and economists Pedro Rosa Dias and Ara Darzi at Imperial College London have calculated that speeding up the development of COVID-19 vaccines by one month would avert the loss of 720,000 years of life and prevent 40 million years in poverty, mostly in lower-income countries.

But Deming thinks that challenge trials should wait until their value is clearer and the risks can be better mitigated, for instance by deploying more potent therapies. €œWe don’t yet know enough low cost vibramycin about this disease to say for this person. You will not die,” she says.

€œWe’ve learned low cost vibramycin so much in the past nine months. In a year, we will be able to do this safely.” This article is reproduced with permission and was first published on October 20 2020.For the first time ever, a NASA probe has performed a sample-snagging operation on an asteroid in deep space. The agency’s OSIRIS-REx spacecraft spiraled down to the surface of the near-Earth asteroid Bennu this afternoon (Oct.

20) to grab material that mission team members hope harbors clues about the solar system’s early days low cost vibramycin and the rise of life on Earth. €œWe did it!. € OSIRIS-REx principal investigator Dante Lauretta, of the University of Arizona, said during a webcast that provided updates about today’s maneuver.

€œWe tagged the surface of low cost vibramycin the asteroid, and it’s up to Bennu now to see how the event went.” The goal was to collect at least 60 grams (2.1 ounces) of dirt and gravel from Bennu’s rubbly surface. It could take up to 10 days to determine if OSIRIS-REx achieved this aim, mission team members have said. And it’s not a low cost vibramycin disaster if the asteroid haul turns out to be a little light.

The probe can go back down for two more tries if need be. “This amazing first for NASA demonstrates how an incredible team from across the country came together and persevered through incredible challenges to expand the boundaries of knowledge,” NASA Administrator Jim Bridenstine said in a statement after the touchdown. €œOur industry, academic, and international partners have made it possible to hold a piece of the most ancient solar system in our hands.” Lauretta and his fellow OSIRIS-REx scientists and engineers watched over today’s asteroid sample-snatching attempt from low cost vibramycin a mission operations center at Lockheed Martin Space in Littleton, Colorado.

(Lockheed Martin built the spacecraft for NASA.) And while the mood was certainly jubilant, the impact of the ongoing COVID-19 pandemic was clear. For example, everyone wore facemasks and maintained appropriate social distancing for much of the event. While there were some hugs after news of OSIRIS-REx’s asteroid touchdown, they were few in low cost vibramycin NASA’s live webcast, with hand sanitizer clearly on hand after such celebrations.

“This is one of those moments where we’re all aware of COVID-19,” NASA astronomer Michelle Thaller of the Goddard Space Flight Center said in the webcast just after touchdown. €œBecause I want the hugs and the high fives and everything, but we’re all going low cost vibramycin to keep each other safe.” The $800 million OSIRIS-REx mission launched in September 2016 and arrived at the 1,640-foot-wide (500 meters) Bennu in December 2018. The probe has been taking the asteroid’s measure ever since, mapping its surface in incredible detail to prepare for today’s maneuver.

That work has revealed a world far more rugged than the mission team had expected. House-sized boulders stud Bennu’s surface, low cost vibramycin limiting the available options for a safe sample grab. The team eventually homed in on a small crater called Nightingale as its top choice, because the site sports relatively fresh and fine-grained material that hasn’t been exposed to the harsh deep-space environment for long.

But Nightingale is surrounded by hazards, including a big outcrop the mission team nicknamed “Mount Doom.” There are obstacles within the crater as well, so the spacecraft targeted a relatively flat, boulder-free area just 26 feet (8 m) wide—quite an ambitious goal, considering that OSIRIS-REx is the size of a 15-passenger van and the original mission plan envisioned a touchdown zone 165 feet (50 m) wide. €œSo, for some perspective low cost vibramycin. The next time you park your car in front of your house or in front of a coffee shop and walk inside, think about the challenge of navigating OSIRIS-REx into one of these spots from 200 million miles away,” Mike Moreau, OSIRIS-REx deputy project manager at NASA’s Goddard Space Flight Center in Greenbelt, Maryland, said during a news conference last month.

It currently low cost vibramycin takes more than 18 minutes for commands to travel from Earth to OSIRIS-REx, so Moreau and his colleagues cannot control the probe in real time. The craft therefore performed today’s operation autonomously. Shortly before 2 p.m.

EDT (1600 GMT) today, OSIRIS-REx fired its thrusters to get out of orbit around Bennu and head low cost vibramycin down toward the surface. At 6:12 p.m. EDT (2212 GMT), the probe “kissed” the asteroid for low cost vibramycin about 10 seconds with its sample-collecting mechanism, which is affixed to the end of OSIRIS-REx’s 11-foot-long (3.4 m) robotic arm.

During the brief touchdown, the spacecraft blasted Bennu’s surface with nitrogen gas. This stirred up dirt and rock that could then be collected by the arm’s sampling head, which mission team members have likened to an older car’s air filter. We should expect OSIRIS-REx’s first images of the operation to start coming down to Earth tomorrow low cost vibramycin morning (Oct.

21), mission team members said. The OSIRIS-REx team will spend the next week or so assessing how much asteroid material was collected. The probe’s handlers have expressed confidence that this low cost vibramycin first attempt will succeed.

OSIRIS-REx’s sampler was designed to snag at least 150 grams (5.3 ounces) and could theoretically get up to 4 kilograms (8.8 lbs.) of material if everything went perfectly. But if OSIRIS-REx is deemed to have come up short on collected low cost vibramycin material today, another attempt could be made, at a backup site known as Osprey, as soon as January 2021. A third try would be possible, too, if needed.

The probe carries three bottles of surface-disturbing nitrogen gas. Those are contingency low cost vibramycin plans, however. If things went according to plan today, OSIRIS-REx remains on course to depart Bennu in March 2021.

The collected samples are scheduled to land here on Earth, encased in a special return capsule, in September 2023. Scientists will then study the material in labs around the world, scrutinizing the stuff in far more detail than OSIRIS-REx, or any other single probe, could do on its own low cost vibramycin in deep space. Asteroids are building blocks left over from the planet-formation epoch, so such analyses could reveal key insights about our solar system’s very early days, NASA officials have said.

€œThis was an incredible feat—and today we’ve advanced both science and engineering and our prospects for future missions to study low cost vibramycin these mysterious ancient storytellers of the solar system,” said Thomas Zurbuchen, NASA’s associate administrator for science missions, in the NASA statement. €œA piece of primordial rock that has witnessed our solar system’s entire history may now be ready to come home for generations of scientific discovery, and we can’t wait to see what comes next.” In addition, Bennu is rich in hydrated minerals and carbon-containing organic compounds. Asteroids like it may have helped Earth become habitable long ago, seeding our planet with the ingredients needed for life as we know it.

€œAnd also, having the samples back here on Earth allows us to preserve them for future generations to come and allows for future explorers to analyze the samples using techniques and instruments that haven’t been invented, and to ask questions that we don’t even know to ask yet,” Lori Glaze, director of NASA’s Planetary Science Division, said during low cost vibramycin today’s webcast. Getting these samples down to Earth is OSIRIS-REx’s top priority. But the mission also has other goals, as indicated by its full name—“Origins, Spectral Interpretation, Resource Identification, Security, Regolith Explorer.” For example, observations the probe has made while orbiting Bennu should help scientists better understand how asteroids move through space, NASA officials have said.

This information low cost vibramycin could improve trajectory projections for potentially hazardous asteroids, a category that includes Bennu. (There’s a 1-in-2,700 chance that Bennu will hit Earth during a close approach in the late 2100s, researchers say.) OSIRIS-REx’s sample won’t be the first pristine asteroid material brought down to Earth by a space mission. Japan’s Hayabusa probe returned some grains of the stony asteroid Itokawa in 2010, and its low cost vibramycin successor, Hayabusa2, recently grabbed pieces of the carbon-rich rock Ryugu.

The material from Ryugu is scheduled to land on Earth this December. The OSIRIS-REx and Hayabusa2 teams have been working together for the past few years, and that collaboration will continue after the missions’ samples touch down on Earth, NASA officials have stressed. Copyright 2020 low cost vibramycin Space.com, a Future company.

All rights reserved. This material may not be published, broadcast, rewritten or redistributed..

Back To Top