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Date published levitra price increase walmart. October 7, 2020On this page OverviewAs the global erectile dysfunction treatment levitra emerged in December 2019, the need for coherent, pan-Canadian guidance on provincial and territorial testing was quickly recognized. Led by the National Microbiology Laboratory, initial interim guidance on laboratory testing was developed in consultation with the Canadian Public Health Lab Network and was finalized and approved by the Special Advisory Committee on April 16, 2020.

This guidance was based on scientific evidence and testing resources available at that levitra price increase walmart time. The recommended testing guidance focused on the molecular polymerase chain reaction (PCR) as the sole laboratory technique to accurately identify erectile dysfunction in a patient sample.In May 2020, based on new evidence, the National Laboratory Testing Indication Guidance for erectile dysfunction treatment was updated to reflect developments in four areas. Expanded laboratory resources viral transmission from asymptomatic individuals or individuals in the pre-symptomatic phase outbreaks in congregate living and work settings new testing modalities (molecular Point of Care and serological tests)The erectile dysfunction treatment landscape has further evolved and it is now necessary to update key aspects of this document to reflect recent scientific and public health data.

One key consideration relates to limiting asymptomatic diagnostic PCR testing where public levitra price increase walmart health action could have significant benefits. Several pilot programs were conducted in Canada, confirming very low levels of erectile dysfunction treatment in the general population and supporting an evidence-based approach to the relaunch of economic activity. In addition, it enabled jurisdictions to stress-test testing capacity and prepare jurisdictions for higher testing volumes.

Asymptomatic testing was also found to levitra price increase walmart displace diagnostic capacity for symptomatic individuals, close contacts, high-risk settings and outbreak management. The National Laboratory Testing Indication Guidancefor erectile dysfunction treatment has been updated to reflect these learnings and advances in science.Recognizing that testing regimes are within provincial and territorial jurisdiction, this document reflects the collaboration among jurisdictions, leveraging learnings from one another through the different adopted approaches.Emerging testing and screening technologiesThe Pan-Canadian erectile dysfunction treatment Testing and Screening Guidance is designed to reflect changing risk management approaches as the levitra conditions change. Recognizing that one size does not fit all, the Guidance is also designed to respond to a significant increase in the need to access testing and screening technologies.

Scaling to meet increased and sustained testing and screening demand will require a paradigm shift, broadening the technologies that are used in a manner that is tailored to the purpose and application of technologies in levitra price increase walmart a variety of settings. Although PCR remains the gold standard in diagnostic testing, numerous technologies and testing modalities are emerging that could serve to supplement diagnostic testing. These recent testing and sampling options could create opportunities to expand the approach to testing by including broad-based approaches to screening through less sensitive and potentially more cost-effective technologies, thereby alleviating strain on the overall public health system.While they can be less sensitive, these technologies could have multiple benefits including ease and reduced cost of production, improved efficiency and reduced reliance on PCR testing supplies.

They also have the potential to be less levitra price increase walmart invasive depending on the technology. Antigen and extraction-free nucleic acid testing are examples of such technologies that, in addition to being more cost-effective and easier to produce, are also easily adaptable to mobile, rapid applications. However, due to their lower sensitivity than current PCR technology, these emerging technologies may be better used as a part of screening, in conjunction with repeated testing in some settings.

Recognizing that these novel technologies have lower sensitivity and specificity than current PCR technology, their use should be targeted to scenarios where both positive and negative are interpreted and acted upon appropriately.Complementing the deployment of these emerging technologies, techniques levitra price increase walmart such as pooled testing are being used to contribute to the preservation of testing resources. Governments are also tapping non-traditional data sources to complement case data. For example, data for wastewater testing could complement erectile dysfunction treatment surveillance systems by providing readily accessible pooled community samples and data for communities where testing is not available or underutilized.As of September 29, Health Canada has authorized 36 erectile dysfunction treatment testing devices (PCR and serological).

Health Canada is fast-tracking the review of submissions related to antigen and levitra price increase walmart nucleic acid tests. Submissions that are reviewed include various sample types, including saliva. Consult the list of authorized medical devices for uses related to erectile dysfunction treatment.In anticipation of regulatory approval for antigen tests, an Interim Guidance on Antigen Testing has been developed to outline potential scenarios such as routine outbreak monitoring, monitoring in different situations including high-risk settings (for example, long-term care facilities) and possible adaptation into mobile, rapid testing in rural and remote communities.Pan-Canadian erectile dysfunction treatment Testing and Screening GuidanceLike the Laboratory Testing Guidance, the Pan-Canadian erectile dysfunction treatment Testing and Screening Guidance (“Guidance”) is based on new public health evidence and emerging technologies, while adopting a broadened approach that leverages and tailors technologies to appropriate uses.

The Guidance is designed to protect and expand the resilience of federal, provincial and territorial testing and screening capacity.The Guidance is based on a portfolio approach that uses different types of levitra price increase walmart testing technologies for various purposes (diagnostic, screening, surveillance). The intent of the Guidance is to better use testing resources to target the most relevant test in particular situations or use cases to address specific problems or purposes. Figure 1.

Technology streams of levitra price increase walmart Pan-Canadian erectile dysfunction treatment Testing and Screening Guidance Figure 1. Technology streams of Pan-Canadian erectile dysfunction treatment Testing and Screening Guidance - Text equivalent Testing. Definitive diagnosis of erectile dysfunction treatment with high sensitivity PCR-based tests, with potential refinements to specimen collecting modalities (for example, saliva) Less amenable to high frequency conduct due to greater resource utilization Screening.

Indicative of erectile dysfunction treatment status, with lower sensitivity levitra price increase walmart Typically newer, rapid technology approaches Amenable to higher frequency repetition and more easily scalable Surveillance. Use of traditional and non-traditional data sources to complement case data Wastewater surveillance complements conventional erectile dysfunction treatment surveillance systems by providing. efficient pooled community sample data for communities where timely clinical testing is underutilized or unavailable data at the local level Five key foundational, interrelated pillars support the advancement of the Guidance.

Scientific integrity regulatory excellence proactive procurement robust data and levitra price increase walmart capacity strategic communication and partnershipsUpdates to laboratory testing and antigen testing guidance founded on rigorous scientific integrity enable and inform decision-making on testing allocations within Canada, and support jurisdictions in the timely use of emerging technologies once regulatory approval is received. Regulatory excellence is equally important as a foundational pillar to implementing the Guidance in a manner that allows for rapid approvals while still preserving the scientific integrity of the process.In addition, undertaking a proactive procurement approach ensures steady access to equipment and supplies for testing and screening. Governments continue to take a proactive procurement approach, purchasing whenever possible, contingent on regulatory approvals.Timely and comprehensive data is critical, underpinning decision-making by governments.

Governments have established a new data set for erectile dysfunction treatment cases that provides more targeted information, improving the ability to understand whether s are acquired via domestic or international travel, or if they are linked to levitra price increase walmart a known outbreak. Race and ethnicity indicators have been added as well as greater information on health care workers, allowing a better understanding of the erectile dysfunction treatment experience among different population groups. In addition to the case data, key data on turnaround times for testing and contact tracing, for example, can also help identify issues related to capacity and timeliness of interventions.Finally, in addition to strong federal, provincial and territorial partnerships, relationships are being further enhanced with key partners in industry and the scientific community.

While ensuring rapid and effective progress is critical, it is levitra price increase walmart also important to communicate what we know, what we are doing and what we are going to do. This collaboration and transparency supports critical decisions, including what additional capacity may be required as part of the Guidance, for instance, federal surge capacity to supplement provincial and territorial leadership. Strategic communications and partnerships are critical to maintaining and strengthening the confidence of Canadians in Governments' actions to address erectile dysfunction treatment.

Implementation plan of the levitra price increase walmart Pan-Canadian erectile dysfunction treatment Testing and Screening Guidance. Updated Guidance Scientific integrity Regulatory excellence Proactive procurement Robust data and capacity Strategic communications and partnerships Regularly updated public health advice as science evolves Updated national lab testing indication guidance Interim antigen testing guidance Guidance on sample types Prioritized, timely review of emerging and promising technologies Responsive to testing, screening and surveillance developments Founded in and driven by scientific excellence Linking regulatory pipeline with production capacity Prioritizing made in Canada solutions Advance purchasing of promising technologies Surge capacity through full value chain and timely, comprehensive data Improving national performance data (turnaround times) Surge capacity for sample collection, lab testing contact tracing Working closely with key partners FPT. Enables agile responses to emerging issues Industry.

Linking public health and workforce requirements Tapping emerging tech Public education/understanding Looking forwardThe Guidance is expected to levitra price increase walmart evolve as the state of knowledge and risk management strategies continue to develop. Guidance on sample types is expected to be finalized during the fall and the balance of testing and screening technologies will be adjusted to respond to the needs of various populations. Researchers and companies continue to innovate and develop new technologies and solutions.

Guidance will need to keep levitra price increase walmart pace with, and take advantage of, these innovations. The continuous updating of this Guidance will rely on strong federal, provincial and territorial partnerships and collaboration leveraging key governance bodies, including the Special Advisory Committee. The Guidance will also capitalize on opportunities to leverage input and the capacity to mobilize knowledge in Canada and from around the world.Related links.

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Statement Health Canada has directory completed its levitra 10mg onsite inspection of the Emergent BioSolutions facility in Baltimore, Maryland. November 24, 2021 | Ottawa, ON | Health Canada Health Canada has completed its onsite inspection of the Emergent BioSolutions facility in Baltimore, Maryland. The European Medicines Agency and levitra 10mg South African Health Products Regulatory Authority also participated in this inspection remotely. All three regulators found the facility to be compliant with Good Manufacturing Practices (GMPs).

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Health Canada’s compliant rating means that Janssen Inc. Will be able to import their treatments into Canada that are made with the drug substance manufactured at the Emergent levitra 10mg BioSolutions facility. While there are no further shipments of the Janssen treatment planned at this time, the Government of Canada will continue working with provinces and territories to identify any further doses that may be required. As with all treatments, each lot of the Janssen erectile dysfunction treatment that could potentially be imported into Canada or donated on Canada’s behalf will be assessed to confirm that it meets Health Canada’s stringent safety and quality requirements.

Only treatment lots that meet these requirements will levitra 10mg be released onto the Canadian market or provided as a Canadian donation. The shipment of Janssen treatments that Canada received on November 10 came from compliant manufacturing sites in Europe. ContactsMedia RelationsHealth Canada613-957-2983media@hc-sc.gc.caStatement As November is Lung levitra 10mg Cancer Awareness Month, it provides a helpful reminder that there are steps you can take to reduce your risk of developing lung cancer. Eat healthy, get more physical activity, and quit smoking.

November 22, 2021 | Ottawa, ON | Health Canada Tobacco use is the leading preventable cause of premature death and disease in Canada. It plays a role in causing more than 40 diseases and other serious health outcomes, levitra 10mg with smoking being the leading cause of lung cancer. Close to 30,000 Canadians every year are diagnosed with lung cancer, a devastating, and often preventable disease. The more, and the longer you smoke, the greater your chance of developing this disease, with nearly 80% of lung cancer cases levitra 10mg attributable to smoking cigarettes.

Exposure to second-hand smoke also increases your risk of developing lung cancer. There is also evidence that people who smoke may be at a greater risk of developing more severe symptoms of erectile dysfunction treatment, as levitra 10mg smoking negatively affects the function of the lungs. Additionally, certain underlying health conditions caused by smoking, such as chronic lung disease and lung cancer, increase the risk of more severe outcomes of erectile dysfunction treatment. Canada’s Tobacco Strategy continues to demonstrate success in tobacco control, including decreasing the smoking rates in Canada to 13%.

Youth rates are at their lowest level in levitra 10mg decades. While this is a step in the right direction, there is more to be done to help those communities that are disproportionately impacted by high rates of smoking, such as Indigenous populations, LGBTQ populations, those suffering with mood or anxiety disorders, as well as people living in poverty. We are taking a levitra 10mg comprehensive and people-centred approach to preventing the death and disease caused by smoking in our society. The Tobacco Products Regulations (Plain and Standardized Appearance) were adopted in 2019 to protect young persons and others from using tobacco products and the consequent dependence on them, and to prevent the public from being deceived or misled with respect to the health hazards of using tobacco products.

As part of Canada’s Tobacco Strategy, we are continuing to invest in public education campaigns and developing new regulations. In May 2021, we announced $3 million to fund a levitra 10mg national social marketing campaign to encourage people who smoke to quit. This campaign is a collaborative effort between the Canadian Cancer Society, the Canadian Lung Association, the Canadian Public Health Association, and the Heart &. Stroke Foundation of Canada levitra 10mg.

The Healthy Canadians and Communities Fund was relaunched in March 2021, and will continue to fund projects that address the behavioural risk factors for chronic disease and will create physical and social environments that are known to support better health among Canadians. This program supports Canadians who face health inequalities and are at greater risk of developing chronic diseases, specifically young males, Indigenous, LGBTQ+ and low income communities. As November is Lung Cancer Awareness Month, it provides a helpful reminder that levitra 10mg there are steps you can take to reduce your risk of developing lung cancer. Eat healthy, get more physical activity, and quit smoking.

Visit Canada.ca/quit-smoking for information and levitra 10mg resources to help you quit smoking. The Honourable Jean-Yves Duclos, P.C., M.P.The Honourable Carolyn Bennett, P.C., M.P. Contacts Marie-France ProulxOffice of Jean-Yves DuclosMinister of Health613-957-0200 Ani DergalstanianOffice of Carolyn BennettMinister of Mental Health and Addictions613-957-0200 Media RelationsHealth Canada613-957-2983media@hc-sc.gc.ca.

Statement Health Canada has completed its onsite inspection of the Emergent levitra price increase walmart BioSolutions facility in Baltimore, Maryland article. November 24, 2021 | Ottawa, ON | Health Canada Health Canada has completed its onsite inspection of the Emergent BioSolutions facility in Baltimore, Maryland. The European Medicines Agency and South African Health Products Regulatory Authority also participated in this inspection remotely levitra price increase walmart. All three regulators found the facility to be compliant with Good Manufacturing Practices (GMPs).

GMPs are an internationally recognized quality assurance system used to ensure that drugs and treatments are made, packaged, levitra price increase walmart labelled, tested, stored, imported and distributed using consistent standards. In June 2021, Health Canada communicated that Canada would not accept any product or ingredients made at the Emergent BioSolutions facility until the Department completed an onsite inspection. This facility manufactures the drug substance used in some doses of the Janssen (Johnson &. Johnson) erectile dysfunction treatment levitra price increase walmart.

Health Canada’s compliant rating means that Janssen Inc. Will be able to import their treatments into Canada levitra price increase walmart that are made with the drug substance manufactured at the Emergent BioSolutions facility. While there are no further shipments of the Janssen treatment planned at this time, the Government of Canada will continue working with provinces and territories to identify any further doses that may be required. As with all treatments, each lot of the Janssen erectile dysfunction treatment that could potentially be imported into Canada or donated on Canada’s behalf will be assessed to confirm that it meets Health Canada’s stringent safety and quality requirements.

Only treatment lots that meet these requirements will be levitra price increase walmart released onto the Canadian market or provided as a Canadian donation. The shipment of Janssen treatments that Canada received on November 10 came from compliant manufacturing sites in Europe. ContactsMedia RelationsHealth Canada613-957-2983media@hc-sc.gc.caStatement As November is Lung Cancer Awareness Month, it provides a helpful reminder that there are steps you can take levitra price increase walmart to reduce your risk of developing lung cancer. Eat healthy, get more physical activity, and quit smoking.

November 22, 2021 | Ottawa, ON | Health Canada Tobacco use is the leading preventable cause of premature death and disease in Canada. It plays a role in causing more than 40 diseases and other serious health levitra price increase walmart outcomes, with smoking being the leading cause of lung cancer. Close to 30,000 Canadians every year are diagnosed with lung cancer, a devastating, and often preventable disease. The more, and the longer you smoke, the greater your chance of developing levitra price increase walmart this disease, with nearly 80% of lung cancer cases attributable to smoking cigarettes.

Exposure to second-hand smoke also increases your risk of developing lung cancer. There is also evidence that people who smoke may be at a greater risk of developing more severe symptoms of erectile dysfunction treatment, as smoking negatively affects levitra price increase walmart the function of the lungs. Additionally, certain underlying health conditions caused by smoking, such as chronic lung disease and lung cancer, increase the risk of more severe outcomes of erectile dysfunction treatment. Canada’s Tobacco Strategy continues to demonstrate success in tobacco control, including decreasing the smoking rates in Canada to 13%.

Youth rates levitra price increase walmart are at their lowest level in decades. While this is a step in the right direction, there is more to be done to help those communities that are disproportionately impacted by high rates of smoking, such as Indigenous populations, LGBTQ populations, those suffering with mood or anxiety disorders, as well as people living in poverty. We are taking a comprehensive and people-centred approach to preventing the death and disease caused by smoking levitra price increase walmart in our society. The Tobacco Products Regulations (Plain and Standardized Appearance) were adopted in 2019 to protect young persons and others from using tobacco products and the consequent dependence on them, and to prevent the public from being deceived or misled with respect to the health hazards of using tobacco products.

As part of Canada’s Tobacco Strategy, we are continuing to invest in public education campaigns and developing new regulations. In May 2021, we announced $3 million to fund a national social marketing campaign levitra price increase walmart to encourage people who smoke to quit. This campaign is a collaborative effort between the Canadian Cancer Society, the Canadian Lung Association, the Canadian Public Health Association, and the Heart &. Stroke Foundation levitra price increase walmart of Canada.

The Healthy Canadians and Communities Fund was relaunched in March 2021, and will continue to fund projects that address the behavioural risk factors for chronic disease and will create physical and social environments that are known to support better health among Canadians. This program supports Canadians who face health inequalities and are at greater risk of developing chronic diseases, specifically young males, Indigenous, LGBTQ+ and low income communities. As November is Lung Cancer Awareness Month, it provides a helpful reminder that there are steps you can take to reduce your risk levitra price increase walmart of developing lung cancer. Eat healthy, get more physical activity, and quit smoking.

Visit Canada.ca/quit-smoking for levitra price increase walmart information and resources to help you quit smoking. The Honourable Jean-Yves Duclos, P.C., M.P.The Honourable Carolyn Bennett, P.C., M.P. Contacts Marie-France ProulxOffice of Jean-Yves DuclosMinister of Health613-957-0200 Ani DergalstanianOffice of Carolyn BennettMinister of Mental Health and Addictions613-957-0200 Media RelationsHealth Canada613-957-2983media@hc-sc.gc.ca.

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Do not take vardenafil if you are taking the following medications:

• nitroglycerin-type drugs for the heart or chest pain such as amyl nitrite, isosorbide dinitrate, isosorbide mononitrate, nitroglycerin, even if these are only taken occasionally. This includes some recreational drugs called 'poppers' which also contain amyl nitrate and butyl nitrate.

Vardenafil may also interact with the following medications:

• alpha blockers such as alfuzosin (UroXatral®), doxazosin (Cardura®), prazosin (Minipress®), tamsulosin (Flomax®), or terazosin (Hytrin®), used to treat high blood pressure or an enlarged prostate.
• arsenic trioxide
• bosentan
• certain antibiotics such as clarithromycin, erythromycin, sparfloxacin, troleandomycin
• certain medicines used for seizures such as carbamazepine, phenytoin, and phenobarbital
• certain medicines for the treatment of HIV or AIDS
• certain medicines to control the heart rhythm (e.g., amiodarone, disopyramide, dofetilide, flecainide, ibutilide, quinidine, procainamide, propafenone, sotalol)
• chloroquine
• cisapride
• diltiazem
• grapefruit juice
• medicines for fungal s (fluconazole, itraconazole, ketoconazole, voriconazole)
• methadone
• nicardipine
• pentamidine
• pimozide
• rifabutin, rifampin, or rifapentine
• some medicines for treating depression or mood problems (amoxapine, maprotiline, fluoxetine, fluvoxamine, nefazodone, pimozide, phenothiazines, tricyclic antidepressants)
• verapamil

Tell your prescriber or health care professional about all other medicines you are taking, including non-prescription medicines, nutritional supplements, or herbal products. Also tell your prescriber or health care professional if you are a frequent user of drinks with caffeine or alcohol, if you smoke, or if you use illegal drugs. These may affect the way your medicine works. Check with your health care professional before stopping or starting any of your medicines.

Can you drink alcohol with levitra

IntroductionIn recent years, many studies have been published on new diagnostic possibilities and management can you drink alcohol with levitra 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 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 can you drink alcohol with levitra 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 countries with similarly can you drink alcohol with levitra 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 can you drink alcohol with levitra 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 can you drink alcohol with levitra 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 team can you drink alcohol with levitra 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 revision, an interdisciplinary can you drink alcohol with levitra 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 can you drink alcohol with levitra 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 terms (eg, DSD, genetic, review, can you drink alcohol with levitra 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 articles that were not open access can you drink alcohol with levitra 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 points of discussion, revised into a can you drink alcohol with levitra 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 to the can you drink alcohol with levitra 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 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 can you drink alcohol with levitra 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 can you drink alcohol with levitra 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 Y chromosomes can you drink alcohol with levitra 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 can you drink alcohol with levitra only be noticed later in pregnancy and 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 the limitations of can you drink alcohol with levitra this technique. The procedure(s) that can be followed, including the risks associated with an amniocentesis.

And the can you drink alcohol with levitra type of information 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 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 can you drink alcohol with levitra 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 with multiple malformations, can you drink alcohol with levitra 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 knowing exactly what the child’s genitalia will look like and uncertainty about the diagnosis, treatment and prognosis cannot be avoided can you drink alcohol with levitra 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 couples decline prenatal testing (personal experience IALG, MK, ABD, YvB, MC and can you drink alcohol with levitra 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 can you drink alcohol with levitra sex assignment is pending.An algorithm 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 can you drink alcohol with levitra 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 can you drink alcohol with levitra before the expected delivery.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." 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 levitra online diagnostic possibilities and management approaches levitra price increase walmart 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 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 levitra price increase walmart 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 levitra price increase walmart 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 Dutch-Flemish guideline that have levitra price increase walmart 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 levitra price increase walmart 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 team requires optimal levitra price increase walmart 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 levitra price increase walmart would benefit from 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 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 levitra price increase walmart. 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 levitra price increase walmart in English and were identified using a broad range of Medical Subject Headings 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 levitra price increase walmart were 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 levitra price increase walmart obtained agreement on remaining 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 levitra price increase walmart incorporating their input, 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 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 levitra price increase walmart 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 levitra price increase walmart in the Netherlands and to all pregnant women in Belgium, and later ultrasound findings. NIPT screens for CNVs in the fetus.

However, depending levitra price increase walmart on legal restrictions 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 DSD is levitra price increase walmart 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 this technique levitra price increase walmart. The procedure(s) that can be followed, including the risks associated with an amniocentesis.

And the type of information genetic levitra price increase walmart 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 geneticist should be experienced in prenatal counselling and well levitra price increase walmart 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 with multiple malformations, but when the DSD occurs as an levitra price increase walmart 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 knowing exactly what the child’s genitalia will look levitra price increase walmart 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 levitra price increase walmart having 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 levitra price increase walmart 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 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 levitra price increase walmart particularly common.27 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 levitra price increase walmart limit, and we propose completing the analysis well before the expected delivery.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." 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, http://www.ec-cath-bischheim.ac-strasbourg.fr/representants/ 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..

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Medicare Part D, the difference between viagra cialis levitra outpatient prescription drug benefit for Medicare beneficiaries, provides coverage above a catastrophic threshold for high out-of-pocket drug costs, but there is no cap on total out-of-pocket drug costs that beneficiaries pay each year. Part D enrollees are required to pay 5% of their total drug costs in the catastrophic phase unless they qualify for Part D Low-Income Subsidies (LIS). In 2021, the catastrophic threshold is set at $6,550 in out-of-pocket drug costs, which includes what beneficiaries themselves pay and difference between viagra cialis levitra the value of the manufacturer discount on the price of brand-name drugs in the coverage gap (sometimes called the “donut hole”), which counts towards this amount. This lack of a hard out-of-pocket spending cap can expose Part D enrollees to thousands of dollars in out-of-pocket costs if they take several costly medications or even just one expensive drug.President Biden has endorsed adding a hard cap on out-of-pocket Medicare Part D prescription drug spending, and this proposed change has also been included in legislation sponsored by policymakers on both sides of the aisle, including H.R. 3, which difference between viagra cialis levitra passed the U.S.

House of Representatives in December 2019 and was recently reintroduced in the 117th Congress. H.R. 19, the House GOP prescription bill (a similar version was introduced in the Senate). Bipartisan legislation that passed out of the Senate Finance Committee in the 116th Congress (S. 2543).

And other legislation. Under H.R. 3, out-of-pocket drug spending under Part D would be capped at $2,000 (beginning in 2024), while under the GOP drug price legislation and the 2019 Senate Finance bill, the cap would be set at $3,100 (beginning in 2022). Under each of these proposals, the out-of-pocket cap excludes the value of the manufacturer price discount. A lower cap would help more beneficiaries and provide more out-of-pocket savings than a higher cap, but could mean higher costs for the federal government, plans, and drug manufacturers, depending on the specific features included in these Part D benefit redesign proposals.To inform discussions about the potential impact of this proposal, in a previous KFF analysis, we analyzed how many Part D enrollees without low-income subsidies exceeded the catastrophic coverage threshold annually and over multiple years, taking into account both beneficiary out-of-pocket spending and the value of the manufacturer discount, and we found that 1.5 million enrollees did so in 2019, and close to 3 million did so between 2015 and 2019.

In this analysis, we focus on the potential impact of different out-of-pocket spending caps in terms of how many beneficiaries would be affected and how much they could save. We analyze how many beneficiaries paid more than $2,000 or $3,100 out of their own pockets for their medications in 2019 (excluding the value of manufacturer discounts they may have received), and the magnitude of potential savings for beneficiaries had these caps been in place in 2019. We also analyze the individual drugs for which Part D enrollees incurred average annual out-of-pocket costs in 2019 above the amount of proposed caps. The analysis is based on 2019 Part D claims data (the most current year available) for Part D enrollees without low-income subsidies (LIS) from the Centers for Medicare &. Medicaid Services Chronic Conditions Data Warehouse (see Methods for details).How many Medicare Part D enrollees incurred out-of-pocket drug costs above $2,000 and $3,100 in 2019?.

In 2019, nearly 1 million more Part D enrollees incurred out-of-pocket costs for their medications above $2,000, the proposed out-of-pocket spending limit in H.R. 3, than above $3,100, the proposed out-of-pocket spending limit in the GOP drug legislation and the 2019 Senate Finance Committee bill (Figure 1). Overall, 1.2 million Part D enrollees in 2019 incurred annual out-of-pocket costs for their medications above $2,000, while 0.3 million spent more than $3,100 out of pocket.The number of Medicare Part D enrollees who have annual out-of-pocket costs greater than $2,000 or $3,100 in a future year, when a proposed cap could be implemented, is likely to exceed our estimates that are based on 2019 claims data, considering enrollment growth, rising drug prices for existing drugs, and the availability of new, higher-priced medications covered by Part D. Moreover, while adding an out-of-pocket cap to Part D may affect a relatively small number of enrollees in any given year, it would help a larger share and number over time, as our previous analysis showed. What is the magnitude of potential savings for Part D enrollees with out-of-pocket costs above $2,000 or $3,100 based on proposed spending caps?.

As expected, a $2,000 cap on out-of-pocket spending would generate larger savings than a $3,100 cap. Average out-of-pocket spending was $3,216 among the 1.2 million Part D enrollees with out-of-pocket spending above $2,000 in 2019. These enrollees would have saved $1,216, or 38% of their annual costs, on average, if a $2,000 cap had been in place in 2019, but only $116, or 4%, under a $3,100 cap (Figure 2). (See Table 1 for estimates of the number of Medicare Part D enrollees with out-of-pocket spending above $2,000 and $3,100 in 2019 by state, and estimated savings under proposed Part D spending caps.)Medicare Part D enrollees with higher-than-average out-of-pocket costs could save substantial amounts with an out-of-pocket spending cap. For example, the top 10% of beneficiaries (122,000 enrollees) with average out-of-pocket costs for their medications above $2,000 in 2019 – who spent at least $5,348 – would have saved $3,348 (63%) in out-of-pocket costs with a $2,000 cap and $2,248 (42%) with a $3,100 cap.

The top 1% of beneficiaries with average out-of-pocket costs above $2,000 (12,000 enrollees) – who spent nearly $12,000 or more – would have saved $9,880 (83%) with a $2,000 cap and $8,780 (74%) with a cap of $3,100.Figure 2. Estimated Cost Savings Under Proposed Medicare Part D Out-of-Pocket Spending Caps Could Be Substantial for Some Part D Enrollees with High Out-of-Pocket CostsHow many and which drugs had average out-of-pocket costs in 2019 above proposed spending caps?. In 2019, there were 154 drugs where Medicare Part D enrollees incurred average annual out-of-pocket costs for that one drug alone greater than $2,000, including 108 drugs where average annual out-of-pocket costs exceeded $3,100.While some of these high-priced drugs are treatments for rare diseases that are taken by a relatively small number of Part D enrollees, the out-of-pocket cost for individual patients taking these drugs can be substantial. For example, average out-of-pocket spending was $42,440 for Strensiq, which treats a rare metabolic disease called hypophosphatasia. $15,108 for Takhzyro, a treatment for hereditary angioedema.

And $13,090 for Firdapse, a treatment for Lambert-Eaton myasthenic syndrome (LEMS), a rare muscle disease. It is important to note that these spending estimates do not include additional out-of-pocket costs that users of these 154 drugs incurred for other medications, so the total out-of-pocket cost burden in 2019 for users of these drugs was likely higher, suggesting that the savings associated with proposed caps on out-of-pocket spending would be even greater than the amount associated with a given relatively high-priced drug.Most of these relatively high-priced drugs were used by fewer than 1,000 non-LIS Part D enrollees in 2019, but 15 drugs were used by at least 5,000 enrollees, and these include drugs to treat cancer, multiple sclerosis (MS), and hepatitis C. For most of these drugs, average out-of-pocket costs in 2019 were well over $2,000, and in many cases well over $3,100 (Figure 3).Average out-of-pocket spending for these 15 drugs in 2019 ranged from $2,300 for abiraterone acetate, a prostate cancer drug used by 14,000 non-LIS enrollees, to $5,700 for Jakafi, a treatment for blood cancer used by 8,000 enrollees.Part D enrollees who used one of these 15 drugs for an entire year spent substantially more than the average user. Average spending tends to understate spending incurred by people who take high-priced drugs for an entire year, since the average includes beneficiaries who begin taking medication after the first of the year (for example, those with a new diagnosis mid-year) as well as those who stop taking a drug at some point during the year for various reasons (such as switching medications, or upon their death). For example:Medicare Part D enrollees who used the cancer drug Revlimid for the entire year in 2019 spent nearly $9,000 out of pocket for this drug alone, two-thirds more than the average user, who only filled 7 prescriptions.Beneficiaries who used the MS drug Tecfidera for the full year in 2019 spent around $4,500 out of pocket for this drug, 31% more than the average user, who filled 9 prescriptions.

Beneficiaries with higher-than-average out-of-pocket costs, including those who take expensive medications for an entire year, could achieve substantial savings under proposed spending caps. For example, Part D enrollees who took Revlimid for the entire year in 2019 would have seen savings of close to $7,000 under a $2,000 spending cap, and more than $5,700 with a $3,100 cap in 2019 (Figure 4) – not including potential savings from other drugs they may also have been taking. DiscussionOur analysis shows that close to 1 million more Medicare Part D enrollees would have had their out-of-pocket costs capped in 2019 under a $2,000 out-of-pocket drug spending limit (as under H.R. 3) than a $3,100 limit (as under the GOP bill and the 2019 Senate Finance Committee bill). Under either cap, however, savings could be considerable for Part D enrollees who take high-cost medications for conditions such as cancer and MS.

Deciding on the level of the cap involves tradeoffs, with more enrollees benefitting and higher out-of-pocket cost savings from a lower cap, but with the potential for higher spending by the federal government, plans, and drug manufacturers, depending on the specific features included in the Part D benefit redesign proposal.While proposed legislation to cap out-of-pocket costs under Medicare Part D would help beneficiaries who take several costly medications or even just one high-priced drug, these proposals would not cap drug spending for expensive physician-administered injectable and infused medications that are covered under Medicare Part B. These drugs are subject to a 20% coinsurance, with no cap on out-of-pocket costs. While many Medicare beneficiaries have supplemental coverage, such as employer-sponsored retiree benefits or Medigap, to help pay their share of costs, nearly 6 million beneficiaries lack supplemental coverage and another 26 million are enrolled in Medicare Advantage plans and typically face 20% coinsurance for Part B drugs up to their plan’s out-of-pocket maximum.For example, a Medicare beneficiary who takes aducanumab, the new Alzheimer’s drug priced at $56,000 annually, would face cost-sharing liability of more than $11,000 in a year, according to KFF analysis, unless they have supplemental insurance. Medicare Advantage enrollees would have a portion of their out-of-pocket costs for this drug covered but would need to pay out of pocket up to their plan’s limit for Medicare Part A and B benefits ($7,550 in 2021 for in-network and $11,300 for in-network and out-of-network combined).The number of Medicare Part D enrollees who have annual out-of-pocket costs greater than $2,000 or $3,100 in a future year, when a proposed cap could be implemented, is likely to exceed our estimates that are based on actual claims data for 2019, considering enrollment growth, rising drug prices for existing drugs, and the availability of new, higher-priced medications covered by Part D. These estimates also do not reflect the interactive effects of other provisions being considered in current prescription drug legislation, such as allowing the federal government to negotiate drug prices or Part B and Part D drug price inflation caps, which would also affect out-of-pocket drug spending.Adding an out-of-pocket cap to Part D would protect Part D enrollees with high drug costs, which may affect only a small share of enrollees in any given year but a larger share over time, including those who have persistently high drug costs over multiple years and others who have high costs in one year but not over time.

The outcome of current discussions in Congress about prescription drug legislation has implications for the affordability of prescription drugs among Medicare beneficiaries.Juliette Cubanski and Tricia Neuman are with KFF. Anthony Damico is an independent consultant.This work was supported in part by Arnold Ventures. We value our funders. KFF maintains full editorial control over all of its policy analysis, polling, and journalism activities. This analysis is based on 2019 Medicare Part D claims data from the Centers for Medicare &.

Medicaid Services Chronic Conditions Data Warehouse for Part D enrollees who are not receiving low-income subsidies (LIS). We exclude Part D enrollees receiving full low-income subsidies because they face only modest cost-sharing amounts before the catastrophic coverage phase and no cost sharing for catastrophic coverage, as well as those receiving partial low-income subsidies, who pay 15% coinsurance before the catastrophic coverage phase and modest copayments of no more than $3.70 for generics and $9.20 for brands in the catastrophic phase.For this analysis, we estimated the number of Part D enrollees without low-income subsidies who had average annual out-of-pocket spending for all the medications they took in 2019 above $2,000 and $3,100, as well as the specific drugs where non-LIS Part D enrollees incurred average annual out-of-pocket costs above $2,000 and $3,100 in 2019. Except where noted, we define a Medicare beneficiary as a full-year user when they either have 12 or more drug fills (generally 30-day supplies) or when their annualized prescription medication quantity received exceeds 360 days.The federal government spent $321 more per person for beneficiaries enrolled in Medicare Advantage plans than for those in traditional Medicare in 2019, a gap that amounted to $7 billion in additional spending on the increasingly popular private plans that year, finds a new KFF analysis.The Medicare Advantage spending includes the cost of extra benefits, such as vision, dental and hearing coverage that are funded by rebates and not covered for beneficiaries in traditional Medicare. The extra benefits have likely contributed to years of steady increases in Medicare Advantage enrollment, which reached 22 million in 2019 (36% of all beneficiaries) and 26 million this year (42%).At the same time, Medicare Advantage spending has risen steadily, and is projected to rise to $664 billion by 2029, up from $348 billion this year. Half of the projected increase is due to growth in enrollment, while the remaining half is attributable to growth in federal payments per enrollee, after accounting for inflation.

The projected growth in spending per Medicare Advantage enrollee is driven in part by the expectation that federal bonus payments that plans receive based on their quality ratings will continue to rise.The higher payments for Medicare Advantage — $11,844 per person in Medicare Advantage vs. $11,523 in traditional Medicare in 2019 — have led to higher federal spending than would have occurred under traditional Medicare and higher Medicare Part B premiums paid by all beneficiaries, including those in traditional Medicare.The higher spending is attributed to features of the Medicare Advantage payment system, including how benchmarks for plan payments are set, as well as the risk adjustment process, that is intended to compensate plans more for higher cost enrollees. That has attracted the attention of the Biden Administration, which in its 2022 budget expressed support for reforming payments to private plans as part of efforts to extend the solvency of the Medicare Hospital Insurance Trust Fund and improve affordability for beneficiaries. Additionally, Medicare Advantage plans have come under scrutiny over inaccurate coding practices that contribute to higher risk scores for their enrollees, and higher payments from Medicare.The new KFF analysis finds that if spending per Medicare Advantage enrollee were 2 percent less each year than the amount projected by the Medicare actuaries – a scenario similar to a recommendation made by the federal Medicare Payment Advisory Commission (MedPAC) — then total Medicare spending would be $82 billion lower than projected between 2021 and 2029.Under a different scenario, if the growth in per person spending on beneficiaries in Medicare Advantage were held to the same rate of growth in spending on beneficiaries in traditional Medicare, then total Medicare program spending would be $183 billion lower than projected between 2021 and 2029, the analysis finds.Reducing Medicare Advantage payments from their projected amounts could have uncertain effects on the availability of plans that offer extra benefits for Medicare Advantage enrollees, or plan profits, unless plans are able to lower administrative costs and operate more efficiently.The full analysis, Higher and Faster Growing Spending Per Medicare Advantage Enrollee Adds to Medicare’s Solvency and Affordability Challenges, as well as other data and analyses about Medicare Advantage, can be found at kff.org..

Medicare Part D, the outpatient prescription drug benefit for Medicare beneficiaries, provides coverage above levitra price increase walmart a catastrophic threshold for high out-of-pocket drug costs, but there is no cap on total out-of-pocket drug costs that beneficiaries pay each year. Part D enrollees are required to pay 5% of their total drug costs in the catastrophic phase unless they qualify for Part D Low-Income Subsidies (LIS). In 2021, the catastrophic threshold is set at $6,550 in out-of-pocket drug costs, which includes what beneficiaries themselves pay and the value of the manufacturer discount on the price of brand-name drugs in the coverage gap (sometimes called the “donut hole”), which levitra price increase walmart counts towards this amount.

This lack of a hard out-of-pocket spending cap can expose Part D enrollees to thousands of dollars in out-of-pocket costs if they take several costly medications or even just one expensive drug.President Biden has endorsed adding a hard cap on out-of-pocket Medicare Part D prescription drug spending, and this proposed change has also been included in legislation sponsored by policymakers on both sides of the aisle, including H.R. 3, which levitra price increase walmart passed the U.S. House of Representatives in December 2019 and was recently reintroduced in the 117th Congress.

H.R. 19, the House GOP prescription bill (a similar version was introduced in the Senate). Bipartisan legislation that passed out of the Senate Finance Committee in the 116th Congress (S.

3, out-of-pocket drug spending under Part D would be capped at $2,000 (beginning in 2024), while under the GOP drug price legislation and the 2019 Senate Finance bill, the cap would be set at $3,100 (beginning in 2022). Under each of these proposals, the out-of-pocket cap excludes the value of the manufacturer price discount. A lower cap would help more beneficiaries and provide more out-of-pocket savings than a higher cap, but could mean higher costs for the federal government, plans, and drug manufacturers, depending on the specific features included in these Part D benefit redesign proposals.To inform discussions about the potential impact of this proposal, in a previous KFF analysis, we analyzed how many Part D enrollees without low-income subsidies exceeded the catastrophic coverage threshold annually and over multiple years, taking into account both beneficiary out-of-pocket spending and the value of the manufacturer discount, and we found that 1.5 million enrollees did so in 2019, and close to 3 million did so between 2015 and 2019.

In this analysis, we focus on the potential impact of different out-of-pocket spending caps in terms of how many beneficiaries would be affected and how much they could save. We analyze how many beneficiaries paid more than $2,000 or $3,100 out of their own pockets for their medications in 2019 (excluding the value of manufacturer discounts they may have received), and the magnitude of potential savings for beneficiaries had these caps been in place in 2019. We also analyze the individual drugs for which Part D enrollees incurred average annual out-of-pocket costs in 2019 above the amount of proposed caps.

The analysis is based on 2019 Part D claims data (the most current year available) for Part D enrollees without low-income subsidies (LIS) from the Centers for Medicare &. Medicaid Services Chronic Conditions Data Warehouse (see Methods for details).How many Medicare Part D enrollees incurred out-of-pocket drug costs above $2,000 and $3,100 in 2019?. In 2019, nearly 1 million more Part D enrollees incurred out-of-pocket costs for their medications above $2,000, the proposed out-of-pocket spending limit in H.R.

3, than above $3,100, the proposed out-of-pocket spending limit in the GOP drug legislation and the 2019 Senate Finance Committee bill (Figure 1). Overall, 1.2 million Part D enrollees in 2019 incurred annual out-of-pocket costs for their medications above $2,000, while 0.3 million spent more than $3,100 out of pocket.The number of Medicare Part D enrollees who have annual out-of-pocket costs greater than $2,000 or $3,100 in a future year, when a proposed cap could be implemented, is likely to exceed our estimates that are based on 2019 claims data, considering enrollment growth, rising drug prices for existing drugs, and the availability of new, higher-priced medications covered by Part D. Moreover, while adding an out-of-pocket cap to Part D may affect a relatively small number of enrollees in any given year, it would help a larger share and number over time, as our previous analysis showed.

What is the magnitude of potential savings for Part D enrollees with out-of-pocket costs above $2,000 or $3,100 based on proposed spending caps?. As expected, a $2,000 cap on out-of-pocket spending would generate larger savings than a $3,100 cap. Average out-of-pocket spending was $3,216 among the 1.2 million Part D enrollees with out-of-pocket spending above $2,000 in 2019.

These enrollees would have saved $1,216, or 38% of their annual costs, on average, if a $2,000 cap had been in place in 2019, but only $116, or 4%, under a $3,100 cap (Figure 2). (See Table 1 for estimates of the number of Medicare Part D enrollees with out-of-pocket spending above $2,000 and $3,100 in 2019 by state, and estimated savings under proposed Part D spending caps.)Medicare Part D enrollees with higher-than-average out-of-pocket costs could save substantial amounts with an out-of-pocket spending cap. For example, the top 10% of beneficiaries (122,000 enrollees) with average out-of-pocket costs for their medications above $2,000 in 2019 – who spent at least $5,348 – would have saved $3,348 (63%) in out-of-pocket costs with a $2,000 cap and $2,248 (42%) with a $3,100 cap.

The top 1% of beneficiaries with average out-of-pocket costs above $2,000 (12,000 enrollees) – who spent nearly $12,000 or more – would have saved $9,880 (83%) with a $2,000 cap and $8,780 (74%) with a cap of $3,100.Figure 2. Estimated Cost Savings Under Proposed Medicare Part D Out-of-Pocket Spending Caps Could Be Substantial for Some Part D Enrollees with High Out-of-Pocket CostsHow many and which drugs had average out-of-pocket costs in 2019 above proposed spending caps?. In 2019, there were 154 drugs where Medicare Part D enrollees incurred average annual out-of-pocket costs for that one drug alone greater than $2,000, including 108 drugs where average annual out-of-pocket costs exceeded $3,100.While some of these high-priced drugs are treatments for rare diseases that are taken by a relatively small number of Part D enrollees, the out-of-pocket cost for individual patients taking these drugs can be substantial.

For example, average out-of-pocket spending was $42,440 for Strensiq, which treats a rare metabolic disease called hypophosphatasia. $15,108 for Takhzyro, a treatment for hereditary angioedema. And $13,090 for Firdapse, a treatment for Lambert-Eaton myasthenic syndrome (LEMS), a rare muscle disease.

It is important to note that these spending estimates do not include additional out-of-pocket costs that users of these 154 drugs incurred for other medications, so the total out-of-pocket cost burden in 2019 for users of these drugs was likely higher, suggesting that the savings associated with proposed caps on out-of-pocket spending would be even greater than the amount associated with a given relatively high-priced drug.Most of these relatively high-priced drugs were used by fewer than 1,000 non-LIS Part D enrollees in 2019, but 15 drugs were used by at least 5,000 enrollees, and these include drugs to treat cancer, multiple sclerosis (MS), and hepatitis C. For most of these drugs, average out-of-pocket costs in 2019 were well over $2,000, and in many cases well over $3,100 (Figure 3).Average out-of-pocket spending for these 15 drugs in 2019 ranged from $2,300 for abiraterone acetate, a prostate cancer drug used by 14,000 non-LIS enrollees, to $5,700 for Jakafi, a treatment for blood cancer used by 8,000 enrollees.Part D enrollees who used one of these 15 drugs for an entire year spent substantially more than the average user. Average spending tends to understate spending incurred by people who take high-priced drugs for an entire year, since the average includes beneficiaries who begin taking medication after the first of the year (for example, those with a new diagnosis mid-year) as well as those who stop taking a drug at some point during the year for various reasons (such as switching medications, or upon their death).

For example:Medicare Part D enrollees who used the cancer drug Revlimid for the entire year in 2019 spent nearly $9,000 out of pocket for this drug alone, two-thirds more than the average user, who only filled 7 prescriptions.Beneficiaries who used the MS drug Tecfidera for the full year in 2019 spent around $4,500 out of pocket for this drug, 31% more than the average user, who filled 9 prescriptions. Beneficiaries with higher-than-average out-of-pocket costs, including those who take expensive medications for an entire year, could achieve substantial savings under proposed spending caps. For example, Part D enrollees who took Revlimid for the entire year in 2019 would have seen savings of close to $7,000 under a $2,000 spending cap, and more than $5,700 with a $3,100 cap in 2019 (Figure 4) – not including potential savings from other drugs they may also have been taking.

DiscussionOur analysis shows that close to 1 million more Medicare Part D enrollees would have had their out-of-pocket costs capped in 2019 under a $2,000 out-of-pocket drug spending limit (as under H.R. 3) than a $3,100 limit (as under the GOP bill and the 2019 Senate Finance Committee bill). Under either cap, however, savings could be considerable for Part D enrollees who take high-cost medications for conditions such as cancer and MS.

Deciding on the level of the cap involves tradeoffs, with more enrollees benefitting and higher out-of-pocket cost savings from a lower cap, but with the potential for higher spending by the federal government, plans, and drug manufacturers, depending on the specific features included in the Part D benefit redesign proposal.While proposed legislation to cap out-of-pocket costs under Medicare Part D would help beneficiaries who take several costly medications or even just one high-priced drug, these proposals would not cap drug spending for expensive physician-administered injectable and infused medications that are covered under Medicare Part B. These drugs are subject to a 20% coinsurance, with no cap on out-of-pocket costs. While many Medicare beneficiaries have supplemental coverage, such as employer-sponsored retiree benefits or Medigap, to help pay their share of costs, nearly 6 million beneficiaries lack supplemental coverage and another 26 million are enrolled in Medicare Advantage plans and typically face 20% coinsurance for Part B drugs up to their plan’s out-of-pocket maximum.For example, a Medicare beneficiary who takes aducanumab, the new Alzheimer’s drug priced at $56,000 annually, would face cost-sharing liability of more than $11,000 in a year, according to KFF analysis, unless they have supplemental insurance.

Medicare Advantage enrollees would have a portion of their out-of-pocket costs for this drug covered but would need to pay out of pocket up to their plan’s limit for Medicare Part A and B benefits ($7,550 in 2021 for in-network and $11,300 for in-network and out-of-network combined).The number of Medicare Part D enrollees who have annual out-of-pocket costs greater than $2,000 or $3,100 in a future year, when a proposed cap could be implemented, is likely to exceed our estimates that are based on actual claims data for 2019, considering enrollment growth, rising drug prices for existing drugs, and the availability of new, higher-priced medications covered by Part D. These estimates also do not reflect the interactive effects of other provisions being considered in current prescription drug legislation, such as allowing the federal government to negotiate drug prices or Part B and Part D drug price inflation caps, which would also affect out-of-pocket drug spending.Adding an out-of-pocket cap to Part D would protect Part D enrollees with high drug costs, which may affect only a small share of enrollees in any given year but a larger share over time, including those who have persistently high drug costs over multiple years and others who have high costs in one year but not over time. The outcome of current discussions in Congress about prescription drug legislation has implications for the affordability of prescription drugs among Medicare beneficiaries.Juliette Cubanski and Tricia Neuman are with KFF.

Anthony Damico is an independent consultant.This work was supported in part by Arnold Ventures. We value our funders. KFF maintains full editorial control over all of its policy analysis, polling, and journalism activities.

This analysis is based on 2019 Medicare Part D claims data from the Centers for Medicare &. Medicaid Services Chronic Conditions Data Warehouse for Part D enrollees who are not receiving low-income subsidies (LIS). We exclude Part D enrollees receiving full low-income subsidies because they face only modest cost-sharing amounts before the catastrophic coverage phase and no cost sharing for catastrophic coverage, as well as those receiving partial low-income subsidies, who pay 15% coinsurance before the catastrophic coverage phase and modest copayments of no more than $3.70 for generics and $9.20 for brands in the catastrophic phase.For this analysis, we estimated the number of Part D enrollees without low-income subsidies who had average annual out-of-pocket spending for all the medications they took in 2019 above $2,000 and $3,100, as well as the specific drugs where non-LIS Part D enrollees incurred average annual out-of-pocket costs above $2,000 and $3,100 in 2019.

Except where noted, we define a Medicare beneficiary as a full-year user when they either have 12 or more drug fills (generally 30-day supplies) or when their annualized prescription medication quantity received exceeds 360 days.The federal government spent $321 more per person for beneficiaries enrolled in Medicare Advantage plans than for those in traditional Medicare in 2019, a gap that amounted to $7 billion in additional spending on the increasingly popular private plans that year, finds a new KFF analysis.The Medicare Advantage spending includes the cost of extra benefits, such as vision, dental and hearing coverage that are funded by rebates and not covered for beneficiaries in traditional Medicare. The extra benefits have likely contributed to years of steady increases in Medicare Advantage enrollment, which reached 22 million in 2019 (36% of all beneficiaries) and 26 million this year (42%).At the same time, Medicare Advantage spending has risen steadily, and is projected to rise to $664 billion by 2029, up from $348 billion this year. Half of the projected increase is due to growth in enrollment, while the remaining half is attributable to growth in federal payments per enrollee, after accounting for inflation.

The projected growth in spending per Medicare Advantage enrollee is driven in part by the expectation that federal bonus payments that plans receive based on their quality ratings will continue to rise.The higher payments for Medicare Advantage — $11,844 per person in Medicare Advantage vs. $11,523 in traditional Medicare in 2019 — have led to higher federal spending than would have occurred under traditional Medicare and higher Medicare Part B premiums paid by all beneficiaries, including those in traditional Medicare.The higher spending is attributed to features of the Medicare Advantage payment system, including how benchmarks for plan payments are set, as well as the risk adjustment process, that is intended to compensate plans more for higher cost enrollees. That has attracted the attention of the Biden Administration, which in its 2022 budget expressed support for reforming payments to private plans as part of efforts to extend the solvency of the Medicare Hospital Insurance Trust Fund and improve affordability for beneficiaries.

Additionally, Medicare Advantage plans have come under scrutiny over inaccurate coding practices that contribute to higher risk scores for their enrollees, and higher payments from Medicare.The new KFF analysis finds that if spending per Medicare Advantage enrollee were 2 percent less each year than the amount projected by the Medicare actuaries – a scenario similar to a recommendation made by the federal Medicare Payment Advisory Commission (MedPAC) — then total Medicare spending would be $82 billion lower than projected between 2021 and 2029.Under a different scenario, if the growth in per person spending on beneficiaries in Medicare Advantage were held to the same rate of growth in spending on beneficiaries in traditional Medicare, then total Medicare program spending would be $183 billion lower than projected between 2021 and 2029, the analysis finds.Reducing Medicare Advantage payments from their projected amounts could have uncertain effects on the availability of plans that offer extra benefits for Medicare Advantage enrollees, or plan profits, unless plans are able to lower administrative costs and operate more efficiently.The full analysis, Higher and Faster Growing Spending Per Medicare Advantage Enrollee Adds to Medicare’s Solvency and Affordability Challenges, as well as other data and analyses about Medicare Advantage, can be found at kff.org..

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AbstractA short levitra 20mg canada cut review was carried out to establish the diagnostic characteristics of alveolar dead space fraction What i should buy with symbicort (AVDSf) in the diagnosis of pulmonary embolism (PE). This is calculated from the arterial and end-tidal CO2. Three papers were selected to answer the levitra 20mg canada clinical question. The author, study type, relevant outcomes, results and weaknesses are tabulated.

It is concluded that there is good evidence to support the use of AVDSf within a clinical prediction model to exclude a PE in patients when there is a low pretest probability. However, the specificity is not sufficient to support it as a ‘rule in’ test.AbstractA short cut review was conducted to assess if the use of rocuronium in the ED was associated with a decrease in the provision of postintubation sedation levitra 20mg canada. Four papers were identified that presented the best evidence to answer the question. Again the studies, relevant outcomes, levitra 20mg canada results and weaknesses are tabulated.

All the identified studies were retrospective and there was a plethora of outcome measures used. When compared with suxamethonium, rocuronium was associated with a delayed initiation and reduced dose of postintubation sedation.emergency care systems.

AbstractA short cut review was carried out to establish the diagnostic characteristics of alveolar dead space fraction (AVDSf) in the diagnosis read this article of pulmonary embolism levitra price increase walmart (PE). This is calculated from the arterial and end-tidal CO2. Three papers levitra price increase walmart were selected to answer the clinical question. The author, study type, relevant outcomes, results and weaknesses are tabulated.

It is concluded that there is good evidence to support the use of AVDSf within a clinical prediction model to exclude a PE in patients when there is a low pretest probability. However, the specificity is not sufficient to support it as a ‘rule in’ test.AbstractA short cut review levitra price increase walmart was conducted to assess if the use of rocuronium in the ED was associated with a decrease in the provision of postintubation sedation. Four papers were identified that presented the best evidence to answer the question. Again the studies, relevant outcomes, results and levitra price increase walmart weaknesses are tabulated.

All the identified studies were retrospective and there was a plethora of outcome measures used. When compared with suxamethonium, rocuronium was associated with a delayed initiation and reduced dose of postintubation sedation.emergency care systems.

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Regulatory considerations and claims In Canada, face coverings that are used only to reduce droplets or aerosols passing between individuals are not regulated as medical devices. However, if the product label includes anti-microbial claims, average price of levitra these face coverings become Class I medical devices. Section 25 of the Medical Device Regulations allows for the request of supporting safety, effectiveness and quality information from Class I manufacturers. Limitations to the claims Bacterial Filtration Efficiency (BFE) is a measurement of a medical mask material's resistance to penetration of a specific kind of microbe. A 3 average price of levitra µm (300 nm) in diameter bacteria.

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Information describing potential inhalation exposure to anti-microbial substance particulates that includes at least. intended use pattern (such as frequency, number of uses) summarized test data that fully characterize the amount (mass) and sizes (particle size distribution and mass median aerodynamic diameter - MMAD) of particulates that are shed during the intended use pattern and human inhalation exposure range estimates in terms of mg/L/hr, and mg/kg-bw/day, based on the information in a) and b) Evidence in the form of test reports that support all anti-viral (anti-erectile dysfunction treatment) and/or antimicrobial claims made on the product label. This may average price of levitra include the use of a scientifically justified surrogate levitra(es). Study details such as specific component/materials that were tested in the test samples should be included, and the test samples should be identical to the product. If there are differences in the materials, concentrations or other properties should be provided along with a justification for how they are representative of the final product in spite of these differences (for example, represent worst-case scenario testing).

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Food and Drug Administration regulates face coverings with anti-microbial claims as medical devices. Self-sanitizing claims are detergent claims that are overseen by the Pest Management Regulatory Agency in Canada and the Environmental average price of levitra Protection Agency in the United States. Related links Glossary of terms Face coverings (also known as non-medical masks). Source control masks (to help control an infected wearer from transmitting the levitra to others) that are made from a variety of woven fabrics. Face coverings may be made of different combinations of fabrics, layering sequences and available average price of levitra in diverse shapes.

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They are average price of levitra designed with 3 layers of non-woven materials and meet labelled filtration levels (80% to 100%) using recognized standards. Personal protective equipment (PPE). Personal protective equipment consists of gowns, gloves, masks, facial protection (masks and eye protection, face shields or masks with visor attachment) or respirators. They can be used by health care workers to provide a barrier that will prevent potential exposure to infectious average price of levitra microorganisms. Respirator.

A device that is tested and certified by procedures established by testing and certification agencies recognized by the authority having jurisdiction and is used to protect the user from inhaling a hazardous atmosphere. The most common respirator used in health care is a N95 half-face piece average price of levitra filtering respirator. It's a personal protective device that fits tightly around the nose and mouth of the wearer. It's used to reduce the risk of inhaling hazardous airborne particles and aerosols, including dust particles and infectious agents.From. Health CanadaDate average price of levitra published.

2021-04-07 Health Canada regulates health products, such as drugs and medical devices. We also regulate consumer and commercial products and substances, such as cosmetics, pesticides, tobacco, cannabis and controlled substances. As part average price of levitra of our regulatory activities, we conduct inspections to mitigate risks and protect the health and safety of Canadians. Learn more about what Health Canada does as a regulator. During the erectile dysfunction treatment levitra, we continue to take a risk-based approach to inspections.

Onsite work remains a key tool in helping us fulfill our mandate average price of levitra to deliver essential inspection activities. Health Canada uses remote or virtual tools to complement onsite inspection activities. We're using these tools, where appropriate and without compromising the ability to verify and assess compliance, for all of the products and substances that we regulate. When onsite activities are conducted, Health Canada is implementing appropriate erectile dysfunction treatment mitigation average price of levitra measures in adherence with public health guidance. Along with erectile dysfunction treatment screening self-assessments, such measures include.

practising social distancing practising good respiratory etiquette and hand hygiene equipping inspectors with sanitation supplies, non-medical masks and other required PPE making adjustments for additional provincial, territorial, local and community specific public health guidance, where applicable Health Canada inspectors are governed by applicable acts and regulations and follow procedures referenced in A Guide to Health Canada Inspections. As such, inspectors continue to have average price of levitra the power to enter any place or premises at any reasonable time where. a regulated activity is being conducted or a regulated product, article, device or thing, or relevant document is located Anyone at the place of the inspection is legally required to give the inspector all reasonable assistance. To stay safe and help limit the spread of erectile dysfunction treatment, Health Canada expects that public health guidance and mitigation measures will be followed while the inspector is onsite. Consideration for the health and safety of inspectors and regulated parties is a joint responsibility.

This notice does not cover my latest blog post anti-microbial agents sold separately and applied to face coverings or medical masks prior levitra price increase walmart to use. On this page About masks with anti-microbial substances The erectile dysfunction treatment levitra has created a public health requirement to wear face coverings and medical masks. Face coverings are not classified as medical devices unless there are medical claims or representations. Some mask and face covering medical devices may incorporate or levitra price increase walmart be coated with materials that claim to be anti-microbial. Anti-microbial substances may kill or inhibit the growth of microorganisms.

Some examples of anti-microbial substances include, but are not limited to. Silver copper Nanoform Graphene fabric coatings saltTo date, Health Canada has not received any data levitra price increase walmart that support the safety and effectiveness of these substances when used with masks or face coverings. It is also not known whether these substances improve the performance of medical masks in a measurable way. Regulatory considerations and claims In Canada, face coverings that are used only to reduce droplets or aerosols passing between individuals are not regulated as medical devices. However, if levitra price increase walmart the product label includes anti-microbial claims, these face coverings become Class I medical devices.

Section 25 of the Medical Device Regulations allows for the request of supporting safety, effectiveness and quality information from Class I manufacturers. Limitations to the claims Bacterial Filtration Efficiency (BFE) is a measurement of a medical mask material's resistance to penetration of a specific kind of microbe. A 3 µm (300 nm) in diameter bacteria levitra price increase walmart. Results are reported as percent efficiency and correlate with the ability of the fabric to resist bacterial penetration. Higher BFE percentages in this test indicate better barrier efficiency.

In general, a levitra price increase walmart BFE rating could be interpreted as material filtration efficiency. This measurement is not to be taken in isolation and without a reference to a test method or international standard. To achieve a high level of filtration, anti-microbial non-medical masks should be manufactured from a non-woven polypropylene material. All claims must be supported by evidence and available levitra price increase walmart for review upon request. Safety and effectiveness requirements Medical masks or other personal protective equipment claiming microbial protection should meet the safety and effectiveness requirements described below.

This information must be available for review upon request in the case of MDEL holders. It should be submitted by manufacturers filing an interim order (IO) application or responding to regulatory requests for levitra price increase walmart information. A clear intended use/indications statement for the product along with complete labelling. Labelling includes user manuals, instructions for use (IFU), directions for use (DFU), outer package labelling, promotional material and website links. A detailed description of the list of materials (for example, chemical levitra price increase walmart and popular/trade names) and their technical specifications (for example, physical/chemical properties), used in the manufacture of the mask.

This includes all material constituents added to the mask to impart anti-microbial or anti-viral properties. A full description of how the anti-microbial or anti-viral technology (for example, coatings) is produced and incorporated into, or bonded with, the mask materials, as well as a mechanistic description of the expected anti-microbial action. If the anti-microbial substances levitra price increase walmart are present in nanoform(s), a characterization of those substances (for example, derivitization, layers, platelets, thickness, lateral dimensions, charged sites), including a certificate of analysis showing impurities. Information describing potential inhalation exposure to anti-microbial substance particulates that includes at least. intended use pattern (such as frequency, number of uses) summarized test data that fully characterize the amount (mass) and sizes (particle size distribution and mass median aerodynamic diameter - MMAD) of particulates that are shed during the intended use pattern and human inhalation exposure range estimates in terms of mg/L/hr, and mg/kg-bw/day, based on the information in a) and b) Evidence in the form of test reports that support all anti-viral (anti-erectile dysfunction treatment) and/or antimicrobial claims made on the product label.

This may include the use of a scientifically justified surrogate levitra(es) levitra price increase walmart. Study details such as specific component/materials that were tested in the test samples should be included, and the test samples should be identical to the product. If there are differences in the materials, concentrations or other properties should be provided along with a justification for how they are representative of the final product in spite of these differences (for example, represent worst-case scenario testing). Evidence of biocompatibility (such as non-cytotoxic, non-irritating and non-sensitizing) of the patient contacting materials levitra price increase walmart in their final manufactured product form. Performance data/reports demonstrating that the respirators/masks meet ASTM F2100, EN 14683, EN 149 and GB2626 (or any other standards claimed).

If it is claimed that the mask can be washed, then instructions for washing should be provided. In addition, evidence must be provided that the performance claims made (for example, in 8 above) are maintained after a proposed maximum number of wash levitra price increase walmart cycles as indicated in the device labelling. International activity The U.S. Food and Drug Administration regulates face coverings with anti-microbial claims as medical devices. Self-sanitizing claims are detergent claims that are overseen by the Pest Management Regulatory Agency in Canada and the Environmental Protection levitra price increase walmart Agency in the United States.

Related links Glossary of terms Face coverings (also known as non-medical masks). Source control masks (to help control an infected wearer from transmitting the levitra to others) that are made from a variety of woven fabrics. Face coverings may be made of different combinations of fabrics, layering levitra price increase walmart sequences and available in diverse shapes. They are a sewn mask secured with ties or straps around the head or behind the ears. They are factory-made or made from household items such as scarves or t-shirts.

The fabrics and/or materials used in face coverings are not the same as the ones used in levitra price increase walmart medical masks or respirators. Medical device:  A device within the meaning of the Food and Drugs Act, but does not include any device that is intended for use in relation to animals.  Medical masks. Includes surgical, procedural, isolation and other control devices intended to offer protection to the wearer. They are designed with 3 layers of non-woven materials and meet labelled filtration levels (80% to 100%) using recognized standards levitra price increase walmart. Personal protective equipment (PPE).

Personal protective equipment consists of gowns, gloves, masks, facial protection (masks and eye protection, face shields or masks with visor attachment) or respirators. They can be used by health care workers to provide a levitra price increase walmart barrier that will prevent potential exposure to infectious microorganisms. Respirator. A device that is tested and certified by procedures established by testing and certification agencies recognized by the authority having jurisdiction and is used to protect the user from inhaling a hazardous atmosphere. The most levitra price increase walmart common respirator used in health care is a N95 half-face piece filtering respirator.

It's a personal protective device that fits tightly around the nose and mouth of the wearer. It's used to reduce the risk of inhaling hazardous airborne particles and aerosols, including dust particles and infectious agents.From. Health CanadaDate levitra price increase walmart published. 2021-04-07 Health Canada regulates health products, such as drugs and medical devices. We also regulate consumer and commercial products and substances, such as cosmetics, pesticides, tobacco, cannabis and controlled substances.

As part of our regulatory activities, we conduct inspections to mitigate risks and levitra price increase walmart protect the health and safety of Canadians. Learn more about what Health Canada does as a regulator. During the erectile dysfunction treatment levitra, we continue to take a risk-based approach to inspections. Onsite work remains a key tool in helping us fulfill levitra price increase walmart our mandate to deliver essential inspection activities. Health Canada uses remote or virtual tools to complement onsite inspection activities.

We're using these tools, where appropriate and without compromising the ability to verify and assess compliance, for all of the products and substances that we regulate. When onsite activities are conducted, Health levitra price increase walmart Canada is implementing appropriate erectile dysfunction treatment mitigation measures in adherence with public health guidance. Along with erectile dysfunction treatment screening self-assessments, such measures include. practising social distancing practising good respiratory etiquette and hand hygiene equipping inspectors with sanitation supplies, non-medical masks and other required PPE making adjustments for additional provincial, territorial, local and community specific public health guidance, where applicable Health Canada inspectors are governed by applicable acts and regulations and follow procedures referenced in A Guide to Health Canada Inspections. As such, inspectors continue to have the power to enter any place or premises at any reasonable time where.