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This how to get levitra online year was our most diverse yet. From interactive events with patients to online webinars and workshops in schools – members pulled out all the stops to raise public awareness of biomedical science and demonstrate the value of the profession and its role in the prevention, diagnosis and treatment of s and disease. The IBMS activity fund provided grants of up to £500 for members to develop their biomedical science-related activities and events. The grants were able to be used to support a wide range of activities and resources, including experiments, exhibition space at careers events, marketing and communications materials, incentives and giveaways how to get levitra online. 'Guardians of your health', Manchester University NHS Foundation Trust Manchester Univerisity NHS Foundation Trust invited patients attending appointments to 'spin the wheel' The Diagnostic Cytopathology biomedical science team held the 'Guardians of your health' to celebrate Biomedical Science Day and raise awareness of the work done at Manchester University NHS Foundation Trust.

The day involved a nationwide webinar organised in partnership with Health Education England (HEE) and several teams from the field across the country attended. The event how to get levitra online leads to a great network opportunity for the department working with HEE to develop a training programme for aspiring biomedical scientists to join the department. The trust invited patients attending appointments to play 'spin the wheel'. The game involved spinning a wheel and answering straightforward multiple-choice questions related to biomedical science for a chance to win a prize. Katharine Hayden how to get levitra online FRCPath commented.

"Thank you all for a fabulous day – so great to see how many people you managed to engage in the event. All down to all of your hard work, well done!. " Zara how to get levitra online Pain HR Director said. "Absolutely amazing and enjoying coffee from the mug I won!. " Another attendee commented.

"It was a fantastic interactive method of raising awareness of the work we do and change the how to get levitra online perception that patients have that Biomedical Scientists only stay in the laboratory. It also provided the opportunity for the public to learn what is a biomedical scientist." Biomedical Science Promotional Video, Jersey Victoria Atherton and the team at the Jersey General Hospital, with the help of the government of Jersey, used the funding to produce a video to help promote the profession. [embedded content] Divine Azange, St Germans Academy School Pupils at St Germans Academy became 'scientists for the day' with Divine Azange Divine Azange used his funds to purchase science kits to hold a 'be a biomedical scientist for a day' event for year four school pupils. Eighteen pupils how to get levitra online received a science kit which included a lab coat, pH meter and Petri dishes. Each pupil ran experiments of magnetic cereal and urinalysis testing - they also conducted the UV glow germ experiment, which looked at hand washing.

This was particularly exciting for the pupils and highlighted the importance of handwashing to curb the spread of erectile dysfunction treatment. They finished by looking at the 'SuperLab' at how to get levitra online the heart of the healthcare comic. Pupils then picked a favourite Superlab hero character and had to guess which superhero Divine was. The pupils did a blood grouping picture panel recognition to finish the day with the winners keeping the UV touch used in the glow germ experiment. Divine is passionate about inspiring how to get levitra online the next generation and believes Year 4 was a great year to highlight the importance biomedical science plays in the patient pathway.

Divine told the IBMS. "It was more about raising curiosity and inspiring minds. The attendees enjoyed being dressed in a lab coat and having a how to get levitra online go at doing their own experiments. The importance of iron in cereal was particularly useful and the pupils found the colour change of urinalysis and pH paper exciting!. " Great British Bake Off, Histology, Royal Wolverhampton Wolverhampton NHS trust Royal Wolverhampton held a Biomedical bake-off competition - yum!.

Ready, Set, how to get levitra online Bake!. The Histology department at Wolverhampton Histology swapped Pipettes for whisks as event lead Amy Bednall organised a laboratory bake-off competition!. Wolverhampton Histology Great Biomedical Bake Off was held in the department to promote the IBMS and to set people a challenge of maybe doing something outside their comfort zone, giving them a chance to be creative. The trust intended to show the great work of the department as it how to get levitra online reconfigures its services and raise awareness of the profession. Everyone in the histology department was invited to join in, including all biomedical scientists, laboratory assistants and pathologists.

Attendee Megan Evans commented. "I took part because how to get levitra online I love to be creative and it's great to feel part of something fun. It is so good for team morale, plus everyone loves a bit of cake afterwards!. !.

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Elon Musk on Friday unveiled a coin-sized prototype of a brain implant developed by his startup Neuralink to enable people who are paralyzed to operate smartphones and robotic limbs with their thoughts — and said the company had worked to “dramatically simplify” the device since presenting an earlier version sildenafil vs cialis vs levitra last summer.In an event live-streamed on YouTube to more than 150,000 viewers at one point, the company staged a demonstration in which it trotted out a pig named Gertrude that was said to have had the company’s device implanted in its head two months ago. The live stream showed what Musk claimed to be Gertrude’s real-time brain activity as it sniffed around a pen. At no point, though, did he provide evidence that the signals — rendered in beeps and bright blue wave sildenafil vs cialis vs levitra patterns on screen — were, in fact, emanating from the pig’s brain.A pig presented at a Neuralink demonstration was said to have one of the company’s brain implants in its head. YouTube screenshot“This is obviously sounding increasingly like a Black Mirror episode,” Musk said at one point during the event as he responded affirmatively to a question about whether the company’s implant could eventually be used to save and replay memories. €œThe future’s going to be weird.”advertisement Musk said that in sildenafil vs cialis vs levitra July Neuralink received a breakthrough device designation from the Food and Drug Administration — a regulatory pathway that could allow the company to soon start a clinical trial in people with paraplegia and tetraplegia.

The big reveal came after four former Neuralink employees told STAT that the company’s leaders have long fostered an internal culture characterized by rushed timelines and the “move fast and break things” ethos of a tech company — a pace sometimes at odds with the slow and incremental pace that’s typical of medical device development. Advertisement Friday’s event began, 40 minutes late, with a glossy video about the company’s work — and then panned to Musk, standing in front of a blue curtain beside a gleaming new version of the company’s surgical “sewing machine” robot that could easily have sildenafil vs cialis vs levitra been mistaken for a giant Apple device. Musk described the event as a “product demo” and said its primary purpose was to recruit potential new employees. It was unclear whether the demonstration was taking place at the company’s sildenafil vs cialis vs levitra Fremont, Calif., headquarters or elsewhere. Musk proceeded to reveal the new version of Neuralink’s brain implant, which he said was designed to fit snugly into the top of the skull.

Neuralink’s technological design has changed significantly since its last big update sildenafil vs cialis vs levitra in July 2019. At that time, the company’s brain implant system involved a credit-card sized device designed to be positioned behind the back of a person’s ear, with several wires stretching to the top of the skull. After demonstrating the pig’s brain activity at Friday’s event, Musk showed video footage of a pig walking on a treadmill and said Neuralink’s sildenafil vs cialis vs levitra device could be used to “predict the position of limbs with high accuracy.” That capability would be critical to allowing someone using the device to do something like controlling a prosthetic limb, for example.Neuralink for months has signaled that it initially plans to develop its device for people who are paralyzed. It said at its July 2019 event that it wanted to start human testing by the end of 2020. Receiving the breakthrough device designation sildenafil vs cialis vs levitra from the FDA — designed to speed up the lengthy regulatory process — is a step forward, but it by no means guarantees that a device will receive a green light, either in a short or longer-term time frame.

After Musk’s presentation, a handful of the company’s employees — all wearing masks, but seated only inches apart — joined him to take questions submitted on Twitter or from the small audience in the room.In typical fashion for a man who in 2018 sent a Tesla Roadster into space, Musk didn’t hesitate to use the event to cross-promote his electric car company. Asked whether sildenafil vs cialis vs levitra the Neuralink chip would allow people to summon their Tesla telepathically, Musk responded. €œDefinitely — of course.”Matthew MacDougall, the company’s head neurosurgeon, appearing in scrubs, said the company had so far only implanted its technology into the brain’s cortical surface, the coaster-width layer enveloping the brain, but added that it hoped to go deeper in the future. Still, Musk said sildenafil vs cialis vs levitra. €œYou could solve blindness, you could solve paralysis, you could solve hearing — you can solve a lot just by interfacing with the cortex.”Musk and MacDougall said they hoped to eventually implant Neuralink’s devices — which they referred to on stage simply as “links” — in the deeper structures of the brain, such as in the hypothalamus, which is believed to play a critical role in mental illnesses including depression, anxiety, and PTSD.There were no updates at the event of Neuralink’s research in monkeys, which the company has been conducting in partnership with the University of California, Davis since 2017.

At last July’s event, Musk said — without providing evidence — that a monkey had controlled a computer with its brain.At that same July 2019 event, Neuralink released a preprint paper — published a few months later — that claimed to show that a series of Neuralink electrodes implanted in the brains of rats could record sildenafil vs cialis vs levitra neural signals. Critically, the work did not show where in the brain the implanted electrodes were recording from, for how long they were recording, or whether the recordings could be linked to any of the rats’ bodily movements.In touting Friday’s event — and Neuralink’s technological capabilities — on Twitter in recent weeks, Musk spoke of “AI symbiosis while u wait” and referenced the “matrix in the matrix” — a science-fiction reference about revealing the true nature of reality. The progress the company reported sildenafil vs cialis vs levitra on Friday fell far short of that. Neuralink’s prototype is ambitious, but it has yet to show evidence that it can match up to the brain-machine interfaces developed by academic labs and other companies. Other groups have shown sildenafil vs cialis vs levitra that they can listen in on neural activity and allow primates and people to control a computer cursor with their brain — so-called “read-out” technology — and have also shown that they can use electrical stimulation to input information, such as a command or the heat of a hot cup of coffee, using “write-in” technology.

Neuralink said on Friday that its technology would have both read-out and write-in capabilities.Musk acknowledged that Neuralink still has a long way to go. In closing the event after sildenafil vs cialis vs levitra more than 70 minutes, Musk said. €œThere’s a tremendous amount of work to be done to go from here to a device that is widely available and affordable and reliable.”Following the news this week of what appears to have been the first confirmed case of a erectile dysfunction treatment re, other researchers have been coming forward with their own reports. One in Belgium, another in the Netherlands sildenafil vs cialis vs levitra. And now, one in Nevada.What caught experts’ attention about the case of the 25-year-old Reno man was not that he appears to have contracted erectile dysfunction (the name of the levitra that causes erectile dysfunction treatment) a second time.

Rather, it’s that his second bout was more serious than his first.Immunologists had expected that if the immune response generated after an initial sildenafil vs cialis vs levitra could not prevent a second case, then it should at least stave off more severe illness. That’s what occurred with the first known re case, in a 33-year-old Hong Kong man.advertisement Still, despite what happened to the man in Nevada, researchers are stressing this is not a sky-is-falling situation or one that should result in firm conclusions. They always presumed people would become vulnerable to erectile dysfunction treatment again some time after recovering from an initial case, based on how our immune sildenafil vs cialis vs levitra systems respond to other respiratory levitraes, including other erectile dysfunctiones. It’s possible that these early cases of re are outliers and have features that won’t apply to the tens of millions of other people who have already shaken off erectile dysfunction treatment.“There are millions and millions of cases,” said Michael Mina, an epidemiologist at Harvard’s T.H. Chan School of sildenafil vs cialis vs levitra Public Health.

The real question that should get the most focus, Mina said, is, “What happens to most people?. €advertisement But with more re reports likely to make it into the scientific sildenafil vs cialis vs levitra literature soon, and from there into the mainstream press, here are some things to look for in assessing them.What’s the deal with the Nevada case?. The Reno resident in question first tested positive for erectile dysfunction in April after coming down with a sore throat, cough, and headache, as well as nausea and diarrhea. He got sildenafil vs cialis vs levitra better over time and later tested negative twice. But then, some 48 days later, the man started experiencing headaches, cough, and other symptoms again.

Eventually, he became so sick that he had to be hospitalized and was found to have pneumonia.Researchers sequenced levitra samples from both of his s and found they were different, providing evidence that this was a new distinct from the first. What happens when we get erectile dysfunction treatment in the first sildenafil vs cialis vs levitra case?. Researchers are finding that, generally, people who get erectile dysfunction treatment develop a healthy immune response replete with both antibodies (molecules that can block pathogens from infecting cells) and T cells (which help wipe out the levitra). This is what happens after other viral s.In addition to fending off the levitra the first time, that immune response also creates memories of the levitra, should it try sildenafil vs cialis vs levitra to invade a second time. It’s thought, then, that people who recover from erectile dysfunction treatment will typically be protected from another case for some amount of time.

With other erectile dysfunctiones, protection is thought to last for perhaps sildenafil vs cialis vs levitra a little less than a year to about three years.But researchers can’t tell how long immunity will last with a new pathogen (like erectile dysfunction) until people start getting reinfected. They also don’t know exactly what mechanisms provide protection against erectile dysfunction treatment, nor do they know what levels of antibodies or T cells are required to signal that someone is protected through a blood test. (These are sildenafil vs cialis vs levitra called the “correlates of protection.”) Why do experts expect second cases to be milder?. With other levitraes, protective immunity doesn’t just vanish one day. Instead, it sildenafil vs cialis vs levitra wanes over time.

Researchers have then hypothesized that with erectile dysfunction, perhaps our immune systems might not always be able to prevent it from getting a toehold in our cells — to halt entirely — but that it could still put up enough of a fight to guard us from getting really sick. Again, this is what happens with other respiratory pathogens.And it’s why some researchers actually looked at the sildenafil vs cialis vs levitra Hong Kong case with relief. The man had mild to moderate erectile dysfunction treatment symptoms during the first case, but was asymptomatic the second time. It was a demonstration, experts said, of what you would want your immune system to sildenafil vs cialis vs levitra do. (The case was only detected because the man’s sample was taken at the airport when he arrived back in Hong Kong after traveling in Europe.)“The fact that somebody may get reinfected is not surprising,” Malik Peiris, a virologist at the University of Hong Kong, told STAT earlier this week about the first re.

€œBut the re didn’t cause disease, so that’s the first point.”The Nevada case, then, provides sildenafil vs cialis vs levitra a counterexample to that. What kind of immune response did the person who was reinfected generate initially?. Earlier, we described the robust immune response that most people who have erectile dysfunction treatment seem to sildenafil vs cialis vs levitra mount. But that was a generalization. s and the immune responses they induce in different people are “heterogeneous,” said Sarah Cobey, an epidemiologist sildenafil vs cialis vs levitra and evolutionary biologist at the University of Chicago.Older people often generate weaker immune responses than younger people.

Some studies have also indicated that milder cases of erectile dysfunction treatment induce tamer immune responses that might not provide as lasting or as thorough of a defense as stronger immune responses. The man in Hong Kong, for example, did not generate antibodies to the levitra after his sildenafil vs cialis vs levitra first , at least to the level that could be detected by blood tests. Perhaps that explains why he contracted the levitra again just about 4 1/2 months after recovering from his initial .In the Nevada case, researchers did not test what kind of immune response the man generated after the first case.“ is not some binary event,” Cobey said. And with re, “there’s going sildenafil vs cialis vs levitra to be some viral replication, but the question is how much is the immune system getting engaged?. €What might be broadly meaningful is when people who mounted robust immune responses start getting reinfected, and how severe their second cases are.

Are people who have erectile dysfunction treatment a second sildenafil vs cialis vs levitra time infectious?. As discussed, immune memory can prevent re. If it sildenafil vs cialis vs levitra can’t, it might stave off serious illness. But there’s a third aspect of this, too.“The most important question for re, with the most serious implications for controlling the levitra, is whether reinfected people can transmit the levitra to others,” Columbia University virologist Angela Rasmussen wrote in Slate this week.Unfortunately, neither the Hong Kong nor the Reno studies looked at this question. But if most people who sildenafil vs cialis vs levitra get reinfected don’t spread the levitra, that’s obviously good news.

What happens when people broadly become susceptible again?. Whether it’s six months after the first or nine months or a year or longer, at some point, protection for most people who sildenafil vs cialis vs levitra recover from erectile dysfunction treatment is expected to wane. And without the arrival of a treatment and broad uptake of it, that could change the dynamics of local outbreaks.In some communities, it’s thought that more than 20% of residents have experienced an initial erectile dysfunction treatment case, and are thus theoretically protected from another case for some time. That is still below the point of sildenafil vs cialis vs levitra herd immunity — when enough people are immune that transmission doesn’t occur — but still, the fewer vulnerable people there are, the less likely spread is to occur.On the flip side though, if more people become susceptible to the levitra again, that could increase the risk of transmission. Modelers are starting to factor that possibility into their forecasts.A crucial question for which there is not an answer yet is whether what happened to the man in Reno, where the second case was more severe than the first, remains a rare occurrence, as researchers expect and hope.

As the Nevada researchers wrote, “the generalizability of this finding is unknown.”An advocacy group has asked the Department of Defense to investigate what it called “an apparent failure” by Moderna (MRNA) sildenafil vs cialis vs levitra to disclose millions of dollars in awards received from the Defense Advanced Research Projects Agency in patent applications the company filed for treatments.In a letter to the agency, Knowledge Ecology International explained that a review of dozens of patent applications found the company received approximately $20 million from the federal government in grants several years ago and the funds “likely” led to the creation of its treatment technology. This was used to develop treatments to combat different levitraes, such as Zika and, later, the levitra that causes erectile dysfunction treatment.In arguing for an investigation, the advocacy group maintained Moderna is obligated under federal law to disclose the grants that led to nearly a dozen specific patent applications and explained the financial support means the U.S. Government would have certain rights sildenafil vs cialis vs levitra over the patents. In other words, U.S. Taxpayers would have sildenafil vs cialis vs levitra an ownership stake in treatments developed by the company.advertisement “This clarifies the public’s right in the inventions,” said Jamie Love, who heads Knowledge Ecology International, a nonprofit that tracks patents and access to medicines issues.

€œThe disclosure (also) changes the narrative about who has financed the inventive activity, often the most risky part of development.” advertisement One particular patent assigned to Moderna concerns methods and compositions that can be used specifically against erectile dysfunctiones, including erectile dysfunction treatment. The patent names a Moderna scientist and a former Moderna scientist as inventors, both of which acknowledged performing work under the DARPA awards in two academic papers, according to the report by the advocacy group.The group examined the 126 patents assigned to Moderna or ModernaTx as well as 154 patent applications. €œDespite the evidence that multiple inventions were sildenafil vs cialis vs levitra conceived in the course of research supported by the DARPA awards, not a single one of the patents or applications assigned to Moderna disclose U.S. Federal government funding,” the report stated.We asked Moderna and the Department of Defense for comment and will update you accordingly.The missive to the Department of Defense follows a recent analysis by Public Citizen, another advocacy group, indicating the National Institutes of Health may own mRNA-1273, the Moderna treatment candidate for erectile dysfunction treatment. The advocacy group noted the federal government filed multiple patents covering the treatment and two patent applications, in particular, sildenafil vs cialis vs levitra list federal scientists as co-inventors.The analyses are part of a larger campaign among advocacy groups and others in the U.S.

And elsewhere to ensure that erectile dysfunction treatment medical products are available to poor populations around the world. The concern reflects the unprecedented global demand for therapies and treatments, sildenafil vs cialis vs levitra and a race among wealthy nations to snap up supplies from treatment makers. In the U.S., the effort has focused on the extent to which the federal government has provided taxpayer dollars to different companies to help fund their discoveries. In some cases, advocates argue sildenafil vs cialis vs levitra that federal funding matters because it clarifies the rights that the U.S. Government has to ensure a therapy or treatment is available to Americans on reasonable terms.One example has been remdesivir, the Gilead Sciences (GILD) treatment being given to hospitalized erectile dysfunction treatment patients.

The role played by the U.S sildenafil vs cialis vs levitra. Government in developing remdesivir to combat erectile dysfunctiones involved contributions from government personnel at such agencies as the U.S. Army Medical Research Institute of Infectious Diseases.As for the Moderna treatment, earlier this month, the company was awarded a $1.525 sildenafil vs cialis vs levitra billion contract by the Department of Defense and the Department of Health and Human Services to manufacture and deliver 100 million doses of its erectile dysfunction treatment. The agreement also includes an option to purchase another 400 million doses, although the terms were not disclosed. In announcing the agreement, the government said it would ensure Americans receive the erectile dysfunction treatment at no cost, although they may be charged by health care providers for administering a shot.In this instance, however, Love said the sildenafil vs cialis vs levitra “letter is not about price or profits.

It’s about (Moderna) not owning up to DARPA funding inventions. If the sildenafil vs cialis vs levitra U.S. Wants to pay for all of the development of Moderna’s treatment, as Moderna now acknowledges, and throw in a few more billion now, and an option to spend billions more, it’s not unreasonable to have some transparency over who paid for their inventions.”This is not the first time Moderna has been accused of insufficient disclosure. Earlier this month, Knowledge Ecology International and Public Citizen maintained the company failed to disclose development costs in a $955 million contract awarded by BARDA for its sildenafil vs cialis vs levitra erectile dysfunction treatment. In all, the federal government has awarded the company approximately $2.5 billion to develop the treatment.The coming few weeks represent a crucial moment for an ambitious plan to try to secure erectile dysfunction treatments for roughly 170 countries around the world without the deep pockets to compete for what will be scarce initial supplies.Under the plan, countries that want to pool resources to buy treatments must notify the World Health Organization and other organizers — Gavi, the treatment Alliance, as well as the Coalition for Epidemic Preparedness Innovations — of their intentions by Monday.

That means it’s fish-or-cut-bait time for the so-called COVAX sildenafil vs cialis vs levitra facility.Already, wealthy countries — the United States, the United Kingdom, Japan, Canada, and Australia, among others, as well as the European Union — have opted to buy their own treatment, signing bilateral contracts with manufacturers that have secured billions of doses of treatment already. That raises the possibility that less wealthy countries will be boxed out of supplies.advertisement And yet Richard Hatchett, the CEO of CEPI, insists there is a path to billions of doses of treatment for the rest of the world in 2021. STAT spoke with Hatchett sildenafil vs cialis vs levitra this week. A transcript of the conversation, lightly edited for clarity and length, follows. You said this is a critical time sildenafil vs cialis vs levitra for CEPI.

Can you explain what needs to happen between now and mid-September for this joint purchasing approach to be a success?. Advertisement The critical moment is now for countries to commit to the COVAX facility, because that will enable us to secure ample quantities of treatment and then to be able to convey when that treatment is likely to become available based on current information.What we’re now here asking countries to do is to indicate their intent to participate by sildenafil vs cialis vs levitra Aug. 31, and to make a binding commitment by Sept. 18. And to provide funds in support of that binding commitment by early October.

Our negotiations with companies are already taking place and it will be important for us from a planning purpose that countries indicate their intent to participate.Those binding commitments we think will be sufficient to allow us to then secure the advance purchase agreements, particularly with those companies that don’t have a prior contractual obligation to COVAX. And then obviously, we need the funds to live up to those advance purchase agreements.Is it possible this thing could still fall apart?. There appears to be some concern COVAX has been boxed out by rich countries. There was always a possibility that there wouldn’t be sufficient uptake. But I think we’re very encouraged at this point by the level of commitment, both from countries that would be beneficiaries of the advance market commitment — that’s the lower-income, lower-middle-income countries — as well as the self-financing countries.

To have over 170 countries expressing interest in participating — they see the value.We’re much more encouraged now that it’s not going to fall apart. We still need to bring it off to maximize its value. And we’re right at the crunch moment where countries are going to have to make these commitments. So, the next month is really absolutely critical to the facility. I am confident at this point that the world recognizes the value and wants it to work.I’ve been keeping tabs on advance purchase agreements that have been announced.

And at this point, a small number of rich countries have nailed down a lot of treatment — more than 3 billion doses. How hard does that make your job?. The fact that they’re doing it creates anxiety among other countries. And that in itself can accelerate the pace. So, I’m not going to say that we’re not watching that with concern.I will say that for COVAX and the facility, this is absolutely critical moment.

I think we still have a window of opportunity between now and mid-September — when we’re asking that the self-financing countries to make their commitments — to make the facility real and to make it work. Between doses that are committed to COVAX through the access agreements and other agreements — these are discussions with partners that CEPI has funded as well as partners that CEPI has not funded — we still see a pathway for COVAX to well over 3 billion doses in 2021.I think it’s really important to bear in mind is that there are at least a few countries — and I think the U.S. And the U.K. Most publicly — that may be in a situation of significant oversupply. I believe the U.S.

And U.K. Numbers, if you add them together, would result in enough treatment for 600 million people to receive two doses of treatment each. And, you know, there is no possible way that the U.S. Or the U.K. Can use that much treatment.So, there may be a lot of extra supply that looks like it’s been tied up sloshing around later.

I don’t think that the bilateral deals that have been struck are going to prevent COVAX from achieving its goals.But if so much treatment has been pre-ordered by rich countries, can countries in the COVAX pool get enough for their needs?. One of the things that we’ve argued through COVAX is that to control the levitra or to end the acute phase of the levitra to allow normalcy to start to reassert itself, you don’t have to vaccinate 100% of your population.You need to vaccinate those at greatest risk for bad outcomes and you need to vaccinate certain critical workers, particularly your health care workforce. And if you can achieve that goal, which for most countries means vaccinating between 20% and maybe 30% of the population, then you can transform the levitra into something that is much more manageable. Then you can buy yourself time to vaccinate everybody who wants to be vaccinated.We’ve argued the COVAX facility really offers the world the best shot at doing that globally in the fastest possible way, as well as providing for equitable access. This is a case where doing the equitable thing is also doing the efficient thing.CEPI has provided funding to nine treatments.

Is it true that all those manufacturers aren’t required to provide the COVAX facility with treatment?. That is correct. One of the things that we did, and I think it was an important role that CEPI played early on, was that we moved money very, very quickly, in small increments. You know, some of the early contracts were only $5 million or $10 million, to get programs up and running while we potentially put in place much larger-scale, longer-term contracts.If you were doing it over again, would you have given money without strings attached?. Yes, I think I would have.

I think that was critically important to initiating programs.Our contract with Moderna was established in about 48 hours. And that provided critical funding to them to manufacture doses that got them into clinical trials within nine weeks of the genetic sequences [of the erectile dysfunction levitra] being released.And if you look at the nine programs that we’ve invested in, seven are in clinical trials. Two — the AstraZeneca program now and the Moderna program — are among the handful in Phase 3 clinical trials. And, I think the number of projects that that we funded initially, which started in kind of a biotech or academic phase that have now been picked up by large multinational corporations, there’s at least four. The Themis program being picked up by Merck, Oxford University by AstraZeneca, the University of Queensland by CSL, and Clover being in partnership with GSK, I think that speaks to the quality of the programs that we selected.So, I think that combination of rapid review, speed of funding, getting those programs started, getting them oriented in the right direction, I think all of that is critical to where we are now.Companies that got money from CEPI to build out production capacity — that money came with strings attached, right?.

Yes, exactly. So, where CEPI has made investments that create manufacturing, or secure manufacturing capacity, the commitment has been that the capacity that is attributable to the CEPI investment is committed — at least right of first refusal — to the global procurement facility.WASHINGTON — The Trump administration removed a top Food and Drug Administration communications official from her post on Friday in the wake of several controversial agency misstatements, a senior administration official confirmed to STAT.The spokeswoman, Emily Miller, had played a lead role in defending the FDA commissioner, Stephen Hahn, after he misrepresented data regarding the use of blood plasma from recovered erectile dysfunction treatment patients. The New York Times first reported Miller’s ouster. Miller’s tenure at as the top FDA spokeswoman lasted only 11 days. Her appointment was viewed with alarm by agency officials who felt her presence at the agency was emblematic of broader political pressure from the Trump administration, STAT first reported earlier this week.advertisement Before joining the FDA, Miller had no experience in health or medicine.

Her former role as assistant commissioner for media affairs is typically not an appointment filled by political appointees. The FDA’s communications arm typically maintains a neutral, nonpolitical tone.Miller’s appointment particularly alarmed FDA staff and outside scientists given her history in right-wing political advocacy and conservatism journalism. Her résumé included a stint as a Washington Times columnist, where she penned columns with titles that include “New Obamacare ads make young women look like sluts,” and a 2013 book on gun rights titled “Emily Gets Her Gun. But Obama Wants to Take Yours.”advertisement She also worked as a reporter for One America News Network, a right-wing cable channel that frequently espouses conspiracy theories and has declared an open alliance with President Trump.Miller quickly made her presence known at the FDA. In the wake of Hahn’s misstatements on blood plasma, she aggressively defended the commissioner, falsely claiming in a tweet that the therapy “has shown to be beneficial for 35% of patients.” An FDA press release on blood plasma, issued less than a week after her appointment, similarly alarmed agency insiders by trumpeting the emergency authorization as “Another Achievement in Administration’s Fight Against [the] levitra.”.

Elon Musk on Friday unveiled a coin-sized prototype of a brain implant developed by his startup Neuralink to enable people who are paralyzed to operate smartphones and robotic limbs with their thoughts — and said the company had worked to “dramatically simplify” the device since presenting an earlier version last summer.In an event live-streamed on YouTube to more than 150,000 viewers at one point, the company staged a demonstration in which it trotted out a pig named Gertrude that was said to have had how to get levitra online the can you buy levitra at walmart company’s device implanted in its head two months ago. The live stream showed what Musk claimed to be Gertrude’s real-time brain activity as it sniffed around a pen. At no point, though, did he provide evidence that the signals — rendered in beeps and bright blue wave patterns on screen — were, in fact, emanating from the pig’s brain.A pig presented at a Neuralink demonstration was said how to get levitra online to have one of the company’s brain implants in its head. YouTube screenshot“This is obviously sounding increasingly like a Black Mirror episode,” Musk said at one point during the event as he responded affirmatively to a question about whether the company’s implant could eventually be used to save and replay memories.

€œThe future’s going to be weird.”advertisement Musk said that in July Neuralink received a breakthrough device designation from the Food and Drug Administration — a regulatory pathway that could allow the company to soon start a clinical trial in people with paraplegia and how to get levitra online tetraplegia. The big reveal came after four former Neuralink employees told STAT that the company’s leaders have long fostered an internal culture characterized by rushed timelines and the “move fast and break things” ethos of a tech company — a pace sometimes at odds with the slow and incremental pace that’s typical of medical device development. Advertisement Friday’s event began, 40 minutes late, with a how to get levitra online glossy video about the company’s work — and then panned to Musk, standing in front of a blue curtain beside a gleaming new version of the company’s surgical “sewing machine” robot that could easily have been mistaken for a giant Apple device. Musk described the event as a “product demo” and said its primary purpose was to recruit potential new employees.

It was unclear whether the demonstration was taking place at the company’s Fremont, Calif., headquarters how to get levitra online or elsewhere. Musk proceeded to reveal the new version of Neuralink’s brain implant, which he said was designed to fit snugly into the top of the skull. Neuralink’s technological design has changed significantly since its last big how to get levitra online update in July 2019. At that time, the company’s brain implant system involved a credit-card sized device designed to be positioned behind the back of a person’s ear, with several wires stretching to the top of the skull.

After demonstrating the pig’s brain activity at Friday’s event, Musk showed video footage of a pig walking on a treadmill and said Neuralink’s device could be used to “predict the position of limbs with high accuracy.” That capability would be critical to allowing someone using the device to do something like controlling a prosthetic limb, for example.Neuralink for how to get levitra online months has signaled that it initially plans to develop its device for people who are paralyzed. It said at its July 2019 event that it wanted to start human testing by the end of 2020. Receiving the breakthrough device designation from the FDA — designed to speed up the how to get levitra online lengthy regulatory process — is a step forward, but it by no means guarantees that a device will receive a green light, either in a short or longer-term time frame. After Musk’s presentation, a handful of the company’s employees — all wearing masks, but seated only inches apart — joined him to take questions submitted on Twitter or from the small audience in the room.In typical fashion for a man who in 2018 sent a Tesla Roadster into space, Musk didn’t hesitate to use the event to cross-promote his electric car company.

Asked whether the Neuralink chip would allow people to summon their Tesla telepathically, Musk responded how to get levitra online. €œDefinitely — of course.”Matthew MacDougall, the company’s head neurosurgeon, appearing in scrubs, said the company had so far only implanted its technology into the brain’s cortical surface, the coaster-width layer enveloping the brain, but added that it hoped to go deeper in the future. Still, Musk how to get levitra online said. €œYou could solve blindness, you could solve paralysis, you could solve hearing — you can solve a lot just by interfacing with the cortex.”Musk and MacDougall said they hoped to eventually implant Neuralink’s devices — which they referred to on stage simply as “links” — in the deeper structures of the brain, such as in the hypothalamus, which is believed to play a critical role in mental illnesses including depression, anxiety, and PTSD.There were no updates at the event of Neuralink’s research in monkeys, which the company has been conducting in partnership with the University of California, Davis since 2017.

At last July’s event, Musk said — how to get levitra online without providing evidence — that a monkey had controlled a computer with its brain.At that same July 2019 event, Neuralink released a preprint paper — published a few months later — that claimed to show that a series of Neuralink electrodes implanted in the brains of rats could record neural signals. Critically, the work did not show where in the brain the implanted electrodes were recording from, for how long they were recording, or whether the recordings could be linked to any of the rats’ bodily movements.In touting Friday’s event — and Neuralink’s technological capabilities — on Twitter in recent weeks, Musk spoke of “AI symbiosis while u wait” and referenced the “matrix in the matrix” — a science-fiction reference about revealing the true nature of reality. The progress the company reported on Friday fell far short how to get levitra online of that. Neuralink’s prototype is ambitious, but it has yet to show evidence that it can match up to the brain-machine interfaces developed by academic labs and other companies.

Other groups have shown that they can listen in on neural activity and allow primates and people to control a computer cursor with their brain — so-called “read-out” technology — and have also shown that they can use electrical stimulation to input information, such as a command or the heat of how to get levitra online a hot cup of coffee, using “write-in” technology. Neuralink said on Friday that its technology would have both read-out and write-in capabilities.Musk acknowledged that Neuralink still has a long way to go. In closing the event after more than 70 minutes, how to get levitra online Musk said. €œThere’s a tremendous amount of work to be done to go from here to a device that is widely available and affordable and reliable.”Following the news this week of what appears to have been the first confirmed case of a erectile dysfunction treatment re, other researchers have been coming forward with their own reports.

One in how to get levitra online Belgium, another in the Netherlands. And now, one in Nevada.What caught experts’ attention about the case of the 25-year-old Reno man was not that he appears to have contracted erectile dysfunction (the name of the levitra that causes erectile dysfunction treatment) a second time. Rather, it’s that his second bout was more serious than how to get levitra online his first.Immunologists had expected that if the immune response generated after an initial could not prevent a second case, then it should at least stave off more severe illness. That’s what occurred with the first known re case, in a 33-year-old Hong Kong man.advertisement Still, despite what happened to the man in Nevada, researchers are stressing this is not a sky-is-falling situation or one that should result in firm conclusions.

They always presumed people would become vulnerable to erectile dysfunction treatment again some time after recovering how to get levitra online from an initial case, based on how our immune systems respond to other respiratory levitraes, including other erectile dysfunctiones. It’s possible that these early cases of re are outliers and have features that won’t apply to the tens of millions of other people who have already shaken off erectile dysfunction treatment.“There are millions and millions of cases,” said Michael Mina, an epidemiologist at Harvard’s T.H. Chan School how to get levitra online of Public Health. The real question that should get the most focus, Mina said, is, “What happens to most people?.

€advertisement But with more re how to get levitra online reports likely to make it into the scientific literature soon, and from there into the mainstream press, here are some things to look for in assessing them.What’s the deal with the Nevada case?. The Reno resident in question first tested positive for erectile dysfunction in April after coming down with a sore throat, cough, and headache, as well as nausea and diarrhea. He got better over time and later tested negative how to get levitra online twice. But then, some 48 days later, the man started experiencing headaches, cough, and other symptoms again.

Eventually, he became so sick that he had to be hospitalized and was found to have pneumonia.Researchers sequenced levitra samples from both of his s and found they were different, providing evidence that this was a new distinct from the first. What happens when we get erectile dysfunction treatment in how to get levitra online the first case?. Researchers are finding that, generally, people who get erectile dysfunction treatment develop a healthy immune response replete with both antibodies (molecules that can block pathogens from infecting cells) and T cells (which help wipe out the levitra). This is what happens after other viral s.In addition to fending off the levitra the first time, that immune response also how to get levitra online creates memories of the levitra, should it try to invade a second time.

It’s thought, then, that people who recover from erectile dysfunction treatment will typically be protected from another case for some amount of time. With other erectile dysfunctiones, protection is thought to last for how to get levitra online perhaps a little less than a year to about three years.But researchers can’t tell how long immunity will last with a new pathogen (like erectile dysfunction) until people start getting reinfected. They also don’t know exactly what mechanisms provide protection against erectile dysfunction treatment, nor do they know what levels of antibodies or T cells are required to signal that someone is protected through a blood test. (These are how to get levitra online called the “correlates of protection.”) Why do experts expect second cases to be milder?.

With other levitraes, protective immunity doesn’t just vanish one day. Instead, it how to get levitra online wanes over time. Researchers have then hypothesized that with erectile dysfunction, perhaps our immune systems might not always be able to prevent it from getting a toehold in our cells — to halt entirely — but that it could still put up enough of a fight to guard us from getting really sick. Again, this how to get levitra online is what happens with other respiratory pathogens.And it’s why some researchers actually looked at the Hong Kong case with relief.

The man had mild to moderate erectile dysfunction treatment symptoms during the first case, but was asymptomatic the second time. It was a demonstration, experts how to get levitra online said, of what you would want your immune system to do. (The case was only detected because the man’s sample was taken at the airport when he arrived back in Hong Kong after traveling in Europe.)“The fact that somebody may get reinfected is not surprising,” Malik Peiris, a virologist at the University of Hong Kong, told STAT earlier this week about the first re. €œBut the re didn’t cause how to get levitra online disease, so that’s the first point.”The Nevada case, then, provides a counterexample to that.

What kind of immune response did the person who was reinfected generate initially?. Earlier, we described the robust immune response that most people who have erectile dysfunction treatment how to get levitra online seem to mount. But that was a generalization. s and the immune responses they induce in different people are “heterogeneous,” said Sarah Cobey, an epidemiologist and evolutionary biologist how to get levitra online at the University of Chicago.Older people often generate weaker immune responses than younger people.

Some studies have also indicated that milder cases of erectile dysfunction treatment induce tamer immune responses that might not provide as lasting or as thorough of a defense as stronger immune responses. The man in Hong Kong, for example, did not generate antibodies to the levitra after his first , at least to the level that how to get levitra online could be detected by blood tests. Perhaps that explains why he contracted the levitra again just about 4 1/2 months after recovering from his initial .In the Nevada case, researchers did not test what kind of immune response the man generated after the first case.“ is not some binary event,” Cobey said. And with re, “there’s going to be how to get levitra online some viral replication, but the question is how much is the immune system getting engaged?.

€What might be broadly meaningful is when people who mounted robust immune responses start getting reinfected, and how severe their second cases are. Are people who have erectile dysfunction treatment a second time how to get levitra online infectious?. As discussed, immune memory can prevent re. If it can’t, it how to get levitra online might stave off serious illness.

But there’s a third aspect of this, too.“The most important question for re, with the most serious implications for controlling the levitra, is whether reinfected people can transmit the levitra to others,” Columbia University virologist Angela Rasmussen wrote in Slate this week.Unfortunately, neither the Hong Kong nor the Reno studies looked at this question. But if most people how to get levitra online who get reinfected don’t spread the levitra, that’s obviously good news. What happens when people broadly become susceptible again?. Whether it’s six months after the first how to get levitra online or nine months or a year or longer, at some point, protection for most people who recover from erectile dysfunction treatment is expected to wane.

And without the arrival of a treatment and broad uptake of it, that could change the dynamics of local outbreaks.In some communities, it’s thought that more than 20% of residents have experienced an initial erectile dysfunction treatment case, and are thus theoretically protected from another case for some time. That is still below the point of herd immunity — when enough people are immune that transmission doesn’t occur — but still, the fewer vulnerable people there are, the less likely spread is to occur.On the flip side though, if more people become susceptible to the levitra again, how to get levitra online that could increase the risk of transmission. Modelers are starting to factor that possibility into their forecasts.A crucial question for which there is not an answer yet is whether what happened to the man in Reno, where the second case was more severe than the first, remains a rare occurrence, as researchers expect and hope. As the Nevada researchers wrote, “the generalizability of this finding is unknown.”An advocacy group has asked the Department of Defense to investigate what it called “an apparent failure” by Moderna (MRNA) to disclose millions of dollars in awards received from the Defense Advanced Research Projects Agency in patent applications the company filed for treatments.In a letter to the agency, Knowledge Ecology International explained that a review of dozens of patent applications found the company received approximately $20 million from the federal government in grants how to get levitra online several years ago and the funds “likely” led to the creation of its treatment technology.

This was used to develop treatments to combat different levitraes, such as Zika and, later, the levitra that causes erectile dysfunction treatment.In arguing for an investigation, the advocacy group maintained Moderna is obligated under federal law to disclose the grants that led to nearly a dozen specific patent applications and explained the financial support means the U.S. Government would have how to get levitra online certain rights over the patents. In other words, U.S. Taxpayers would have an ownership stake in treatments developed by the company.advertisement “This clarifies the public’s right in the inventions,” said Jamie Love, who heads Knowledge Ecology International, a nonprofit that tracks patents and access to medicines issues how to get levitra online.

€œThe disclosure (also) changes the narrative about who has financed the inventive activity, often the most risky part of development.” check that advertisement One particular patent assigned to Moderna concerns methods and compositions that can be used specifically against erectile dysfunctiones, including erectile dysfunction treatment. The patent names a Moderna scientist and a former Moderna scientist as inventors, both of which acknowledged performing work under the DARPA awards in two academic papers, according to the report by the advocacy group.The group examined the 126 patents assigned to Moderna or ModernaTx as well as 154 patent applications. €œDespite the evidence that multiple inventions how to get levitra online were conceived in the course of research supported by the DARPA awards, not a single one of the patents or applications assigned to Moderna disclose U.S. Federal government funding,” the report stated.We asked Moderna and the Department of Defense for comment and will update you accordingly.The missive to the Department of Defense follows a recent analysis by Public Citizen, another advocacy group, indicating the National Institutes of Health may own mRNA-1273, the Moderna treatment candidate for erectile dysfunction treatment.

The advocacy group noted the federal government filed multiple patents covering the treatment and two patent applications, how to get levitra online in particular, list federal scientists as co-inventors.The analyses are part of a larger campaign among advocacy groups and others in the U.S. And elsewhere to ensure that erectile dysfunction treatment medical products are available to poor populations around the world. The concern reflects the unprecedented global demand for therapies and treatments, and a race among wealthy nations to snap up supplies how to get levitra online from treatment makers. In the U.S., the effort has focused on the extent to which the federal government has provided taxpayer dollars to different companies to help fund their discoveries.

In some cases, advocates argue that federal funding matters because it clarifies the rights that how to get levitra online the U.S. Government has to ensure a therapy or treatment is available to Americans on reasonable terms.One example has been remdesivir, the Gilead Sciences (GILD) treatment being given to hospitalized erectile dysfunction treatment patients. The role played by the U.S how to get levitra online. Government in developing remdesivir to combat erectile dysfunctiones involved contributions from government personnel at such agencies as the U.S.

Army Medical Research how to get levitra online Institute of Infectious Diseases.As for the Moderna treatment, earlier this month, the company was awarded a $1.525 billion contract by the Department of Defense and the Department of Health and Human Services to manufacture and deliver 100 million doses of its erectile dysfunction treatment. The agreement also includes an option to purchase another 400 million doses, although the terms were not disclosed. In announcing the agreement, the government said it would ensure Americans receive the erectile dysfunction treatment at no cost, although they how to get levitra online may be charged by health care providers for administering a shot.In this instance, however, Love said the “letter is not about price or profits. It’s about (Moderna) not owning up to DARPA funding inventions.

If the how to get levitra online U.S. Wants to pay for all of the development of Moderna’s treatment, as Moderna now acknowledges, and throw in a few more billion now, and an option to spend billions more, it’s not unreasonable to have some transparency over who paid for their inventions.”This is not the first time Moderna has been accused of insufficient disclosure. Earlier this month, how to get levitra online Knowledge Ecology International and Public Citizen maintained the company failed to disclose development costs in a $955 million contract awarded by BARDA for its erectile dysfunction treatment. In all, the federal government has awarded the company approximately $2.5 billion to develop the treatment.The coming few weeks represent a crucial moment for an ambitious plan to try to secure erectile dysfunction treatments for roughly 170 countries around the world without the deep pockets to compete for what will be scarce initial supplies.Under the plan, countries that want to pool resources to buy treatments must notify the World Health Organization and other organizers — Gavi, the treatment Alliance, as well as the Coalition for Epidemic Preparedness Innovations — of their intentions by Monday.

That means it’s fish-or-cut-bait time for the so-called COVAX facility.Already, wealthy countries — the United States, the United Kingdom, Japan, Canada, and Australia, among others, as well as the European Union — have opted to buy their how to get levitra online own treatment, signing bilateral contracts with manufacturers that have secured billions of doses of treatment already. That raises the possibility that less wealthy countries will be boxed out of supplies.advertisement And yet Richard Hatchett, the CEO of CEPI, insists there is a path to billions of doses of treatment for the rest of the world in 2021. STAT spoke with how to get levitra online Hatchett this week. A transcript of the conversation, lightly edited for clarity and length, follows.

You said this is a critical time for how to get levitra online CEPI. Can you explain what needs to happen between now and mid-September for this joint purchasing approach to be a success?. Advertisement The how to get levitra online critical moment is now for countries to commit to the COVAX facility, because that will enable us to secure ample quantities of treatment and then to be able to convey when that treatment is likely to become available based on current information.What we’re now here asking countries to do is to indicate their intent to participate by Aug. 31, and to make a binding commitment by Sept.

18. And to provide funds in support of that binding commitment by early October. Our negotiations with companies are already taking place and it will be important for us from a planning purpose that countries indicate their intent to participate.Those binding commitments we think will be sufficient to allow us to then secure the advance purchase agreements, particularly with those companies that don’t have a prior contractual obligation to COVAX. And then obviously, we need the funds to live up to those advance purchase agreements.Is it possible this thing could still fall apart?.

There appears to be some concern COVAX has been boxed out by rich countries. There was always a possibility that there wouldn’t be sufficient uptake. But I think we’re very encouraged at this point by the level of commitment, both from countries that would be beneficiaries of the advance market commitment — that’s the lower-income, lower-middle-income countries — as well as the self-financing countries. To have over 170 countries expressing interest in participating — they see the value.We’re much more encouraged now that it’s not going to fall apart.

We still need to bring it off to maximize its value. And we’re right at the crunch moment where countries are going to have to make these commitments. So, the next month is really absolutely critical to the facility. I am confident at this point that the world recognizes the value and wants it to work.I’ve been keeping tabs on advance purchase agreements that have been announced.

And at this point, a small number of rich countries have nailed down a lot of treatment — more than 3 billion doses. How hard does that make your job?. The fact that they’re doing it creates anxiety among other countries. And that in itself can accelerate the pace.

So, I’m not going to say that we’re not watching that with concern.I will say that for COVAX and the facility, this is absolutely critical moment. I think we still have a window of opportunity between now and mid-September — when we’re asking that the self-financing countries to make their commitments — to make the facility real and to make it work. Between doses that are committed to COVAX through the access agreements and other agreements — these are discussions with partners that CEPI has funded as well as partners that CEPI has not funded — we still see a pathway for COVAX to well over 3 billion doses in 2021.I think it’s really important to bear in mind is that there are at least a few countries — and I think the U.S. And the U.K.

Most publicly — that may be in a situation of significant oversupply. I believe the U.S. And U.K. Numbers, if you add them together, would result in enough treatment for 600 million people to receive two doses of treatment each.

And, you know, there is no possible way that the U.S. Or the U.K. Can use that much treatment.So, there may be a lot of extra supply that looks like it’s been tied up sloshing around later. I don’t think that the bilateral deals that have been struck are going to prevent COVAX from achieving its goals.But if so much treatment has been pre-ordered by rich countries, can countries in the COVAX pool get enough for their needs?.

One of the things that we’ve argued through COVAX is that to control the levitra or to end the acute phase of the levitra to allow normalcy to start to reassert itself, you don’t have to vaccinate 100% of your population.You need to vaccinate those at greatest risk for bad outcomes and you need to vaccinate certain critical workers, particularly your health care workforce. And if you can achieve that goal, which for most countries means vaccinating between 20% and maybe 30% of the population, then you can transform the levitra into something that is much more manageable. Then you can buy yourself time to vaccinate everybody who wants to be vaccinated.We’ve argued the COVAX facility really offers the world the best shot at doing that globally in the fastest possible way, as well as providing for equitable access. This is a case where doing the equitable thing is also doing the efficient thing.CEPI has provided funding to nine treatments.

Is it true that all those manufacturers aren’t required to provide the COVAX facility with treatment?. That is correct. One of the things that we did, and I think it was an important role that CEPI played early on, was that we moved money very, very quickly, in small increments. You know, some of the early contracts were only $5 million or $10 million, to get programs up and running while we potentially put in place much larger-scale, longer-term contracts.If you were doing it over again, would you have given money without strings attached?.

Yes, I think I would have. I think that was critically important to initiating programs.Our contract with Moderna was established in about 48 hours. And that provided critical funding to them to manufacture doses that got them into clinical trials within nine weeks of the genetic sequences [of the erectile dysfunction levitra] being released.And if you look at the nine programs that we’ve invested in, seven are in clinical trials. Two — the AstraZeneca program now and the Moderna program — are among the handful in Phase 3 clinical trials.

And, I think the number of projects that that we funded initially, which started in kind of a biotech or academic phase that have now been picked up by large multinational corporations, there’s at least four. The Themis program being picked up by Merck, Oxford University by AstraZeneca, the University of Queensland by CSL, and Clover being in partnership with GSK, I think that speaks to the quality of the programs that we selected.So, I think that combination of rapid review, speed of funding, getting those programs started, getting them oriented in the right direction, I think all of that is critical to where we are now.Companies that got money from CEPI to build out production capacity — that money came with strings attached, right?. Yes, exactly. So, where CEPI has made investments that create manufacturing, or secure manufacturing capacity, the commitment has been that the capacity that is attributable to the CEPI investment is committed — at least right of first refusal — to the global procurement facility.WASHINGTON — The Trump administration removed a top Food and Drug Administration communications official from her post on Friday in the wake of several controversial agency misstatements, a senior administration official confirmed to STAT.The spokeswoman, Emily Miller, had played a lead role in defending the FDA commissioner, Stephen Hahn, after he misrepresented data regarding the use of blood plasma from recovered erectile dysfunction treatment patients.

The New York Times first reported Miller’s ouster. Miller’s tenure at as the top FDA spokeswoman lasted only 11 days. Her appointment was viewed with alarm by agency officials who felt her presence at the agency was emblematic of broader political pressure from the Trump administration, STAT first reported earlier this week.advertisement Before joining the FDA, Miller had no experience in health or medicine. Her former role as assistant commissioner for media affairs is typically not an appointment filled by political appointees.

The FDA’s communications arm typically maintains a neutral, nonpolitical tone.Miller’s appointment particularly alarmed FDA staff and outside scientists given her history in right-wing political advocacy and conservatism journalism. Her résumé included a stint as a Washington Times columnist, where she penned columns with titles that include “New Obamacare ads make young women look like sluts,” and a 2013 book on gun rights titled “Emily Gets Her Gun. But Obama Wants to Take Yours.”advertisement She also worked as a reporter for One America News Network, a right-wing cable channel that frequently espouses conspiracy theories and has declared an open alliance with President Trump.Miller quickly made her presence known at the FDA. In the wake of Hahn’s misstatements on blood plasma, she aggressively defended the commissioner, falsely claiming in a tweet that the therapy “has shown to be beneficial for 35% of patients.” An FDA press release on blood plasma, issued less than a week after her appointment, similarly alarmed agency insiders by trumpeting the emergency authorization as “Another Achievement in Administration’s Fight Against [the] levitra.”.

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Among people with Medicare, Black beneficiaries are more likely to have cost-related problems with their health care than White beneficiaries, finds a new KFF analysis, with the racial disparity persisting among beneficiaries in both traditional Medicare and Medicare Advantage plans.While 17 percent of all Medicare beneficiaries, or about 1 in 6, reported health care cost-related problems in 2018, the rate among Black beneficiaries buy levitra canada was double that among White beneficiaries (28% vs. 14%), according to the analysis of data from the 2018 Medicare Current Beneficiary Survey (MCBS).Among Medicare Advantage enrollees, the rate of cost-related problems among Black beneficiaries was also higher than among White beneficiaries (32% vs buy levitra canada. 16%), the analysis finds.Among Black beneficiaries specifically, a larger share of those in Medicare Advantage reported cost-related problems buy levitra canada than those in traditional Medicare (32% vs. 24%). The rate of cost-related problems was lower still among the subset of Black beneficiaries in traditional Medicare who had Medicaid or other sources of supplemental insurance (20%).Cost-related problems were defined in buy levitra canada the analysis as trouble getting care due to cost, a delay in care due to cost, or problems paying medical bills.Across all Medicare beneficiaries, a somewhat smaller share of those in traditional Medicare than in Medicare Advantage reported cost-related problems (15% vs.

19%), with a lower rate among beneficiaries buy levitra canada in traditional Medicare with supplemental coverage (12%). The analysis also shows that, overall and across racial and ethnic groups, the Medicare beneficiaries who are most likely to experience cost-related problems are those in traditional Medicare without supplemental coverage – 30 percent of whom reported cost-related problems in 2018.Rates of cost-related problems were even higher among Black beneficiaries in fair or poor self-reported health, where half (50%) of those in Medicare Advantage experienced cost-related problems and one-third (34%) of those in traditional Medicare.The analysis finds that enrollees in Medicare Advantage, who now account for more than 4 in 10 beneficiaries overall, do not generally receive greater protection against cost-related problems than beneficiaries in traditional Medicare with supplemental coverage, despite requirements for Medicare Advantage plans to have buy levitra canada out-of-pocket limits. Differences in cost-related problems between Medicare Advantage and traditional Medicare with supplemental coverage are not fully explained by differences in the characteristics of beneficiaries, such as income and health status.The new findings are noteworthy in that half of all Black beneficiaries are enrolled in Medicare Advantage (compared to just over one third of White beneficiaries).However, the analysis does not estimate actual differences in average out-of-pocket spending among these groups, because it is not possible to derive comparable and accurate estimates of spending for Medicare Advantage enrollees using the Medicare Current Beneficiary Survey, as can be done for traditional Medicare beneficiaries.For more data and analyses about Medicare and racial equity and health policy, visit kff.org.

Among people with Medicare, Black beneficiaries are more likely to have cost-related problems with their health care than blog link White beneficiaries, finds a new KFF analysis, with the racial disparity persisting among beneficiaries in both traditional Medicare and Medicare Advantage plans.While 17 percent of all Medicare beneficiaries, or about 1 in 6, reported health care cost-related problems in 2018, the how to get levitra online rate among Black beneficiaries was double that among White beneficiaries (28% vs. 14%), according to how to get levitra online the analysis of data from the 2018 Medicare Current Beneficiary Survey (MCBS).Among Medicare Advantage enrollees, the rate of cost-related problems among Black beneficiaries was also higher than among White beneficiaries (32% vs. 16%), the analysis finds.Among Black beneficiaries specifically, a larger share of those in Medicare Advantage reported cost-related problems than those in traditional how to get levitra online Medicare (32% vs. 24%). The rate of cost-related problems was lower still among the subset of Black beneficiaries in traditional Medicare who had Medicaid or other sources of supplemental insurance (20%).Cost-related problems were defined in the analysis as trouble getting care due to cost, a delay in care due to how to get levitra online cost, or problems paying medical bills.Across all Medicare beneficiaries, a somewhat smaller share of those in traditional Medicare than in Medicare Advantage reported cost-related problems (15% vs.

19%), with a lower rate among beneficiaries in traditional Medicare with how to get levitra online supplemental coverage (12%). The analysis how to get levitra online also shows that, overall and across racial and ethnic groups, the Medicare beneficiaries who are most likely to experience cost-related problems are those in traditional Medicare without supplemental coverage – 30 percent of whom reported cost-related problems in 2018.Rates of cost-related problems were even higher among Black beneficiaries in fair or poor self-reported health, where half (50%) of those in Medicare Advantage experienced cost-related problems and one-third (34%) of those in traditional Medicare.The analysis finds that enrollees in Medicare Advantage, who now account for more than 4 in 10 beneficiaries overall, do not generally receive greater protection against cost-related problems than beneficiaries in traditional Medicare with supplemental coverage, despite requirements for Medicare Advantage plans to have out-of-pocket limits. Differences in cost-related problems between Medicare Advantage and traditional Medicare with supplemental coverage are not fully explained by differences in the characteristics of beneficiaries, such as income and health status.The new findings are noteworthy in that half of all Black beneficiaries are enrolled in Medicare Advantage (compared to just over one third of White beneficiaries).However, the analysis does not estimate actual differences in average out-of-pocket spending among these groups, because it is not possible to derive comparable and accurate estimates of spending for Medicare Advantage enrollees using the Medicare Current Beneficiary Survey, as can be done for traditional Medicare beneficiaries.For more data and analyses about Medicare and racial equity and health policy, visit kff.org.

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As a world-leading authority http://johnmcdonaghconsulting.com/cheap-levitra/ on can i buy levitra over the counter adult congenital heart disease (ACHD), Professor Michael A. Gatzoulis believes the field is a success story of modern medicine with an ever-increasing number of patients able to reach and enjoy adulthood, including those with complex disease who previously had a guarded prognosis.Yet he also concedes there is still a long way to go to improve the care for this group of patients with life-long disease. In addition to more evidence-based practice for this global can i buy levitra over the counter disease affecting approximately 1% of new-borns, he advocates a personalized ACHD approach with patient education and empowerment at its heart, and better use of technology.A Consultant Cardiologist at the Royal Brompton &.

Harefield NHS Trust in London, and the Academic Head of ACHD and Pulmonary Arterial Hypertension (PAH), his key objective has been ‘to promote the needs of patients with congenital heart disease, including delivery of best care, translation research, training, and education’.Prof. Gatzoulis was born into an ‘open-minded and supportive’ medical family in Drama, a city in the north-east of Greece, where his older brother Konstantinos—currently Professor of Cardiology at the University of Athens—and younger sister Thalia (now a successful artist) also studied medicine. His father, Athanasios, was a paediatrician.‘I was very rebellious as a teenager’, he recalled, ‘my old friends hardly recognize me can i buy levitra over the counter now’.

Indeed, he failed to enter the Aristotelian University of Thessaloniki at the first attempt but was accepted the following year in 1977. Following graduation and his national service as a soldier and a year’s provincial service as a young doctor, he left his homeland for London in 1987 ‘for a new challenge’, securing a paediatric Senior House Officer role working long hours. €˜I liked the environment and the responsibility, can i buy levitra over the counter though in the beginning it was challenging.

For the first few months I was coming home totally exhausted, but it got better, and I have no regrets’.With an interest in the heart, he felt the natural next step was paediatric cardiology, so he seized the opportunity when consultant paediatric cardiologist Dr Michael Rigby asked him to join the Royal Brompton Hospital in 1992 (Figure 1). Figure 1Royal Brompton Hospital, South Block, Circa 1880.Figure 1Royal Brompton can i buy levitra over the counter Hospital, South Block, Circa 1880.With his PhD mentor there, Prof. Andrew Redington, he began studying the right ventricle (RV) in adult patients with Tetralogy of Fallot, which led to several important publications.Right ventricular diastolic function, he added, was linked for the first time to arrhythmia and propensity to sudden cardiac death, the ‘mechano-electric concept’, which had implications on prognostication and led to a proactive approach towards pulmonary valve replacement (Figure 2).1 Figure 2ECG with broad QRS complex and a CMR of a dilated RV with pulmonary regurgitation (PR).

ECG with broad QRS complex and 1st degree heart block of a patient with repaired Tetralogy of Fallot presenting with sustained VT. Note QRS can i buy levitra over the counter >. 180 ms.

Composite shows cardiac MRI from the patient with (A) and (B) moderate to severe pulmonary regurgitation, (C) marked dilatation and can i buy levitra over the counter some hypertrophy of the right ventricle with secondary tricuspid regurgitation and (D) Right pulmonary artery (RPA) stenosis at the site of a previous Blalock–Taussig Shunt. Patient underwent surgical PV implantation, relief of RPA stenosis, and AICD implantation.Figure 2ECG with broad QRS complex and a CMR of a dilated RV with pulmonary regurgitation (PR). ECG with broad QRS complex and 1st degree heart block of a patient with repaired Tetralogy of Fallot presenting with sustained VT.

Note QRS > can i buy levitra over the counter. 180 ms. Composite shows cardiac MRI from the patient with (A) and (B) moderate to severe pulmonary regurgitation, (C) marked dilatation and some hypertrophy of the right ventricle with secondary tricuspid regurgitation and (D) Right pulmonary artery (RPA) stenosis at the site of a previous Blalock–Taussig Shunt.

Patient underwent surgical PV implantation, relief of RPA stenosis, and can i buy levitra over the counter AICD implantation.‘It was a golden era for paediatric cardiology at the Brompton’, said Prof. Gatzoulis, ‘and I could clearly see that ACHD was an area of growth and need’.Having completed his post-graduate training in London by the end of 1996, and smitten by clinical research, he decided not to go back to Greece but moved instead to Canada to work with Gary Webb at the Toronto General Hospital.Returning to London in 1999, he became head of the GUCH (Grown-up CHD) unit at Royal Brompton Hospital, succeeding Prof. Jane Somerville, to run and expand one of the world’s largest ACHD clinical, training and research programmes.Today, Prof.

Gatzoulis is the academic head of the Adult Congenital Heart Centre and the Centre for Pulmonary Hypertension and clinical lead for ACHD at the Royal Brompton Hospital and a Professor of Cardiology and CHD at the National Heart and Lung can i buy levitra over the counter Institute, Imperial College, London. Together with his colleagues, he looks after more than 10 000 ACHD and 1000 PAH patients, including those with the greatest disease complexity.Among his influences was his father, ‘an amazing person ahead of his time’, who after a successful career as a paediatrician retired at 60 and embarked on a new calling on nature preservation and community work. Prof.

Redington taught him how to conduct and report research, while Dr Webb was inspiring with his ‘inclusivity and painstaking work on databases’. But he also reflects on the influence of obstetrician Prof. Phil Steer from the Chelsea &.

Westminster Hospital, ‘for his patient-centred holistic approach and team building, while maintaining academic rigour and output’.Prof. Gatzoulis’ research focus has been on mechanisms and prevention of heart failure (HF) and sudden cardiac death in CHD and the treatment of PAH. He said.

€˜We have improved the outlook for CHD patients a great deal, but for the most part we have not fixed it’. Prof. Gatzoulis recalls how Prof.

Andrew Coats, then head of research at the Royal Brompton, was supportive of his early descriptive work on heart failure markers and exercise intolerance in ACHD, which have now become standard practice (Figure 3). €˜Our work reinforced that we have not cured CHD and at the same time we have opened new therapeutic opportunities’. Figure 3Heart failure and transplantation teams from the Royal Brompton and Harefield NHS Trust at their regular Multi-disciplinary Team (MDT) meeting at the Brompton site.Figure 3Heart failure and transplantation teams from the Royal Brompton and Harefield NHS Trust at their regular Multi-disciplinary Team (MDT) meeting at the Brompton site.The Brompton’s designation as a national centre for ACHD and PAH in 2002 was a significant step forward, delivering greater patient numbers for his team to understand the pathophysiology and try novel therapies.

€˜Patients with Eisenmenger Syndrome (ES), the extreme end of the CHD-PAH spectrum, were either neglected or mismanaged by dogma and we have done a lot of work on pathophysiology of their condition and advanced therapies (Figure 4), which has transformed their lives and relevant practice’. Figure 4Peripheral cyanosis in a patient with ES PDA, Graph showing improved 6 minute-walk-distance (MWD) and survival from disease targeting therapy (DTT), Composite Figure. Right panel.

Peripheral cyanosis. Only possible diagnosis is a Patent Ductus Arteriosus and Eisenmenger Syndrome (ES. Take the patient’s socks off).

Left panel. (A, B) Improvement on pulmonary vascular resistance index (PVRi) and the 6 MWD in patients with ES after 16 weeks of Bosentan therapy versus placebo, BREATHE 5 study, (C) Improvement in symptoms and QoL after 16 weeks of intention to treat patients with ES with iron supplementation and (D) Survival benefit of patients with ES on PAH advanced therapies. From Gatzoulis et al.

IJC 2014, permission granted.Figure 4Peripheral cyanosis in a patient with ES PDA, Graph showing improved 6 minute-walk-distance (MWD) and survival from disease targeting therapy (DTT), Composite Figure. Right panel. Peripheral cyanosis.

Only possible diagnosis is a Patent Ductus Arteriosus and Eisenmenger Syndrome (ES. Take the patient’s socks off). Left panel.

(A, B) Improvement on pulmonary vascular resistance index (PVRi) and the 6 MWD in patients with ES after 16 weeks of Bosentan therapy versus placebo, BREATHE 5 study, (C) Improvement in symptoms and QoL after 16 weeks of intention to treat patients with ES with iron supplementation and (D) Survival benefit of patients with ES on PAH advanced therapies. From Gatzoulis et al. IJC 2014, permission granted.

Figure 5Paul Wood Textbook cover ‘Diseases of the Heart and Circulation’.Figure 5Paul Wood Textbook cover ‘Diseases of the Heart and Circulation’.He believes his original work on pulmonary regurgitation/right ventricular function, with his serendipitous ECG observations, the mechano-electric concept, together with the groundwork on HF and the clinical trials on PAH in the context of CHD, as among his most important.Asked how he thinks his work has advanced the field, Prof. Gatzoulis replied. €˜There was a major impact from our research on ACHD practice with our proactive approach.

A lot of the recent focus of mainstream cardiology—for example, the right ventricle, the pulmonary vascular bed, and even transaortic valve implantation (TAVI)—relate to original research or innovations originating from CHD.‘Overall, we have made progress, but we cannot be complacent. There is clearly more to do. More evidence is needed to inform our practice and we must work more collaboratively to achieve this.

CHD is a very heterogeneous disease. And we are not doing a good enough job in empowering patients to lead independent and full lives.‘Now is the time to move to a more patient-centred, holistic approach, where we are truly the patient’s advocate. Education is central to this and merits further investment.2 Better use of technology, including Artificial Intelligence3 and remote monitoring are also due and have come to the fore due to the self-isolation protocols of the erectile dysfunction treatment levitra'.Prof.

Gatzoulis is particularly proud of the 150+ ACHD Fellows that trained with him at the Brompton. €˜The number one asset for me is the patient, but number two—and close behind—are the Fellows who come to train with us in ACHD. The fact is that I learn from them—and from the patients—more than they learn from me!.

The ACHD Fellows are now all over the world practising ACHD and I am immensely proud of them’.A former president of the International Society for Adult Congenital Heart Disease, a council member of the ACHD WG of the ESC, and recipient of multiple awards including the prestigious Aristotle Medal for the Year for Science and Politics (2019), he is the author of over 380 peer-reviewed publications, edited or co-edited 10 cardiology textbooks, is an incoming Deputy Editor of the EHJ, Associate Editor of the International Journal of Cardiology and is launching a new journal with a CHD and PAH focus.A father of two teenage boys, away from medicine he enjoys tennis, watersports and cycling, food markets/cooking/restaurants, museums/arts, and travel.Advice he would give young researchers as they set out on a path toward success within the field is. €˜Pursue your dreams and seek the right environment. Work hard and stay close to the patient and to your research, and do not be put off by obstacles’.Prof.

Gatzoulis, as one of the leading pioneers, points to future challenges in the ‘continuously evolving cardiovascular subspecialty’ of ACHD. Understanding better the late course of the disease. Optimizing therapies.

Ensuring patient access to tertiary care. Training of the new generation of professionals to serve ACHD patients. And securing resources (Figure 5).4‘Visionary heads of cardiology always had space for this unique cardiovascular subspecialty for the wealth of its anatomic spectrum, the intriguing physiology, the wonderful clinical signs and the deserving patients.

The examples are multiple, from the late Paul Wood at the Brompton, to Eugene Braunwald in Boston, to Pavlos Toutouzas in Athens and many others.‘The number of adult ACHD patients has long exceeded the number of children with CHD’, he said, ‘yet the provision of care for the former is lacking. Furthermore, patients and their families have not been educated and empowered enough regarding their CHD, lifestyle issues and outlook and, yet they navigate their lives with a positive attitude, despite uncertainty, multiple operations, and physical disability in some. For me, the patients are the true heroes in this journey, and a daily inspiration’.

ReferencesReferences are available as supplementary material at European Heart Journal online.Conflict of interest. None declared. Published on behalf of the European Society of Cardiology.

All rights reserved. © The Author(s) 2020. For permissions, please email.

As a world-leading authority on adult congenital heart disease how to get levitra online (ACHD), Professor Michael Cheap levitra A. Gatzoulis believes the field is a success story of modern medicine with an ever-increasing number of patients able to reach and enjoy adulthood, including those with complex disease who previously had a guarded prognosis.Yet he also concedes there is still a long way to go to improve the care for this group of patients with life-long disease. In addition to more evidence-based practice for this how to get levitra online global disease affecting approximately 1% of new-borns, he advocates a personalized ACHD approach with patient education and empowerment at its heart, and better use of technology.A Consultant Cardiologist at the Royal Brompton &. Harefield NHS Trust in London, and the Academic Head of ACHD and Pulmonary Arterial Hypertension (PAH), his key objective has been ‘to promote the needs of patients with congenital heart disease, including delivery of best care, translation research, training, and education’.Prof.

Gatzoulis was born into an ‘open-minded and supportive’ medical family in Drama, a city in the north-east of Greece, where his older brother Konstantinos—currently Professor of Cardiology at the University of Athens—and younger sister Thalia (now a successful artist) also studied medicine. His father, Athanasios, how to get levitra online was a paediatrician.‘I was very rebellious as a teenager’, he recalled, ‘my old friends hardly recognize me now’. Indeed, he failed to enter the Aristotelian University of Thessaloniki at the first attempt but was accepted the following year in 1977. Following graduation and his national service as a soldier and a year’s provincial service as a young doctor, he left his homeland for London in 1987 ‘for a new challenge’, securing a paediatric Senior House Officer role working long hours.

€˜I liked the environment and the responsibility, though in how to get levitra online the beginning it was challenging. For the first few months I was coming home totally exhausted, but it got better, and I have no regrets’.With an interest in the heart, he felt the natural next step was paediatric cardiology, so he seized the opportunity when consultant paediatric cardiologist Dr Michael Rigby asked him to join the Royal Brompton Hospital in 1992 (Figure 1). Figure 1Royal Brompton Hospital, how to get levitra online South Block, Circa 1880.Figure 1Royal Brompton Hospital, South Block, Circa 1880.With his PhD mentor there, Prof. Andrew Redington, he began studying the right ventricle (RV) in adult patients with Tetralogy of Fallot, which led to several important publications.Right ventricular diastolic function, he added, was linked for the first time to arrhythmia and propensity to sudden cardiac death, the ‘mechano-electric concept’, which had implications on prognostication and led to a proactive approach towards pulmonary valve replacement (Figure 2).1 Figure 2ECG with broad QRS complex and a CMR of a dilated RV with pulmonary regurgitation (PR).

ECG with broad QRS complex and 1st degree heart block of a patient with repaired Tetralogy of Fallot presenting with sustained VT. Note QRS how to get levitra online >. 180 ms. Composite shows cardiac MRI from the patient with (A) how to get levitra online and (B) moderate to severe pulmonary regurgitation, (C) marked dilatation and some hypertrophy of the right ventricle with secondary tricuspid regurgitation and (D) Right pulmonary artery (RPA) stenosis at the site of a previous Blalock–Taussig Shunt.

Patient underwent surgical PV implantation, relief of RPA stenosis, and AICD implantation.Figure 2ECG with broad QRS complex and a CMR of a dilated RV with pulmonary regurgitation (PR). ECG with broad QRS complex and 1st degree heart block of a patient with repaired Tetralogy of Fallot presenting with sustained VT. Note QRS > how to get levitra online. 180 ms.

Composite shows cardiac MRI from the patient with (A) and (B) moderate to severe pulmonary regurgitation, (C) marked dilatation and some hypertrophy of the right ventricle with secondary tricuspid regurgitation and (D) Right pulmonary artery (RPA) stenosis at the site of a previous Blalock–Taussig Shunt. Patient underwent surgical how to get levitra online PV implantation, relief of RPA stenosis, and AICD implantation.‘It was a golden era for paediatric cardiology at the Brompton’, said Prof. Gatzoulis, ‘and I could clearly see that ACHD was an area of growth and need’.Having completed his post-graduate training in London by the end of 1996, and smitten by clinical research, he decided not to go back to Greece but moved instead to Canada to work with Gary Webb at the Toronto General Hospital.Returning to London in 1999, he became head of the GUCH (Grown-up CHD) unit at Royal Brompton Hospital, succeeding Prof. Jane Somerville, to run and expand one of the world’s largest ACHD clinical, training and research programmes.Today, Prof.

Gatzoulis is the academic head of the Adult Congenital Heart Centre how to get levitra online and the Centre for Pulmonary Hypertension and clinical lead for ACHD at the Royal Brompton Hospital and a Professor of Cardiology and CHD at the National Heart and Lung Institute, Imperial College, London. Together with his colleagues, he looks after more than 10 000 ACHD and 1000 PAH patients, including those with the greatest disease complexity.Among his influences was his father, ‘an amazing person ahead of his time’, who after a successful career as a paediatrician retired at 60 and embarked on a new calling on nature preservation and community work. Prof. Redington taught him how to conduct and report research, while Dr Webb was inspiring with his ‘inclusivity and painstaking work on databases’.

But he also reflects on the influence of obstetrician Prof. Phil Steer from the Chelsea &. Westminster Hospital, ‘for his patient-centred holistic approach and team building, while maintaining academic rigour and output’.Prof. Gatzoulis’ research focus has been on mechanisms and prevention of heart failure (HF) and sudden cardiac death in CHD and the treatment of PAH.

He said. €˜We have improved the outlook for CHD patients a great deal, but for the most part we have not fixed it’. Prof. Gatzoulis recalls how Prof.

Andrew Coats, then head of research at the Royal Brompton, was supportive of his early descriptive work on heart failure markers and exercise intolerance in ACHD, which have now become standard practice (Figure 3). €˜Our work reinforced that we have not cured CHD and at the same time we have opened new therapeutic opportunities’. Figure 3Heart failure and transplantation teams from the Royal Brompton and Harefield NHS Trust at their regular Multi-disciplinary Team (MDT) meeting at the Brompton site.Figure 3Heart failure and transplantation teams from the Royal Brompton and Harefield NHS Trust at their regular Multi-disciplinary Team (MDT) meeting at the Brompton site.The Brompton’s designation as a national centre for ACHD and PAH in 2002 was a significant step forward, delivering greater patient numbers for his team to understand the pathophysiology and try novel therapies. €˜Patients with Eisenmenger Syndrome (ES), the extreme end of the CHD-PAH spectrum, were either neglected or mismanaged by dogma and we have done a lot of work on pathophysiology of their condition and advanced therapies (Figure 4), which has transformed their lives and relevant practice’.

Figure 4Peripheral cyanosis in a patient with ES PDA, Graph showing improved 6 minute-walk-distance (MWD) and survival from disease targeting therapy (DTT), Composite Figure. Right panel. Peripheral cyanosis. Only possible diagnosis is a Patent Ductus Arteriosus and Eisenmenger Syndrome (ES.

Take the patient’s socks off). Left panel. (A, B) Improvement on pulmonary vascular resistance index (PVRi) and the 6 MWD in patients with ES after 16 weeks of Bosentan therapy versus placebo, BREATHE 5 study, (C) Improvement in symptoms and QoL after 16 weeks of intention to treat patients with ES with iron supplementation and (D) Survival benefit of patients with ES on PAH advanced therapies. From Gatzoulis et al.

IJC 2014, permission granted.Figure 4Peripheral cyanosis in a patient with ES PDA, Graph showing improved 6 minute-walk-distance (MWD) and survival from disease targeting therapy (DTT), Composite Figure. Right panel. Peripheral cyanosis. Only possible diagnosis is a Patent Ductus Arteriosus and Eisenmenger Syndrome (ES.

Take the patient’s socks off). Left panel. (A, B) Improvement on pulmonary vascular resistance index (PVRi) and the 6 MWD in patients with ES after 16 weeks of Bosentan therapy versus placebo, BREATHE 5 study, (C) Improvement in symptoms and QoL after 16 weeks of intention to treat patients with ES with iron supplementation and (D) Survival benefit of patients with ES on PAH advanced therapies. From Gatzoulis et al.

IJC 2014, permission granted. Figure 5Paul Wood Textbook cover ‘Diseases of the Heart and Circulation’.Figure 5Paul Wood Textbook cover ‘Diseases of the Heart and Circulation’.He believes his original work on pulmonary regurgitation/right ventricular function, with his serendipitous ECG observations, the mechano-electric concept, together with the groundwork on HF and the clinical trials on PAH in the context of CHD, as among his most important.Asked how he thinks his work has advanced the field, Prof. Gatzoulis replied. €˜There was a major impact from our research on ACHD practice with our proactive approach.

A lot of the recent focus of mainstream cardiology—for example, the right ventricle, the pulmonary vascular bed, and even transaortic valve implantation (TAVI)—relate to original research or innovations originating from CHD.‘Overall, we have made progress, but we cannot be complacent. There is clearly more to do. More evidence is needed to inform our practice and we must work more collaboratively to achieve this. CHD is a very heterogeneous disease.

And we are not doing a good enough job in empowering patients to lead independent and full lives.‘Now is the time to move to a more patient-centred, holistic approach, where we are truly the patient’s advocate. Education is central to this and merits further investment.2 Better use of technology, including Artificial Intelligence3 and remote monitoring are also due and have come to the fore due to the self-isolation protocols of the erectile dysfunction treatment levitra'.Prof. Gatzoulis is particularly proud of the 150+ ACHD Fellows that trained with him at the Brompton. €˜The number one asset for me is the patient, but number two—and close behind—are the Fellows who come to train with us in ACHD.

The fact is that I learn from them—and from the patients—more than they learn from me!. The ACHD Fellows are now all over the world practising ACHD and I am immensely proud of them’.A former president of the International Society for Adult Congenital Heart Disease, a council member of the ACHD WG of the ESC, and recipient of multiple awards including the prestigious Aristotle Medal for the Year for Science and Politics (2019), he is the author of over 380 peer-reviewed publications, edited or co-edited 10 cardiology textbooks, is an incoming Deputy Editor of the EHJ, Associate Editor of the International Journal of Cardiology and is launching a new journal with a CHD and PAH focus.A father of two teenage boys, away from medicine he enjoys tennis, watersports and cycling, food markets/cooking/restaurants, museums/arts, and travel.Advice he would give young researchers as they set out on a path toward success within the field is. €˜Pursue your dreams and seek the right environment. Work hard and stay close to the patient and to your research, and do not be put off by obstacles’.Prof.

Gatzoulis, as one of the leading pioneers, points to future challenges in the ‘continuously evolving cardiovascular subspecialty’ of ACHD. Understanding better the late course of the disease. Optimizing therapies. Ensuring patient access to tertiary care.

Training of the new generation of professionals to serve ACHD patients. And securing resources (Figure 5).4‘Visionary heads of cardiology always had space for this unique cardiovascular subspecialty for the wealth of its anatomic spectrum, the intriguing physiology, the wonderful clinical signs and the deserving patients. The examples are multiple, from the late Paul Wood at the Brompton, to Eugene Braunwald in Boston, to Pavlos Toutouzas in Athens and many others.‘The number of adult ACHD patients has long exceeded the number of children with CHD’, he said, ‘yet the provision of care for the former is lacking. Furthermore, patients and their families have not been educated and empowered enough regarding their CHD, lifestyle issues and outlook and, yet they navigate their lives with a positive attitude, despite uncertainty, multiple operations, and physical disability in some.

For me, the patients are the true heroes in this journey, and a daily inspiration’. ReferencesReferences are available as supplementary material at European Heart Journal online.Conflict of interest. None declared. Published on behalf of the European Society of Cardiology.

All rights reserved. © The Author(s) 2020. For permissions, please email. Journals.permissions@oup.com..

Buy levitra vardenafil

The transpopulation represents a vulnerable population her latest blog segment both socially and medically, with a buy levitra vardenafil higher incidence of mental health issues. During the erectile dysfunction treatment outbreak, transgender persons have faced additional social, psychological and physical difficulties.1 2 In Italy and in several other buy levitra vardenafil countries access to healthcare has been difficult or impossible thereby hindering the start or continuation of hormonal and psychological treatments. Furthermore, several planned gender-affirming buy levitra vardenafil surgeries have been postponed. These obstacles may have caused an additional psychological burden given the positive effects of medical and surgical treatments on well-being, directly and indirectly, reducing stressors such as workplace discrimination and social inequalities.3 Some organisational aspects buy levitra vardenafil should also be considered.

Binary gender policies may worsen inequalities and marginalisation of transgender subjects potentially increasing the risk of morbidity and mortality.As with the general population, during the lockdown, the Internet and social media were useful in reducing isolation and, in this particular population, were also relevant for buy levitra vardenafil keeping in touch with associations and healthcare facilities with the support of telemedicine services.4 Addressing the role of the telemedicine in the transpopulation, between May and June 2020 we conducted an anonymous web-based survey among transgenders living in Italy (ClinicalTrials.gov Identifier NCT04448418). Among the 108 respondents, with a mean age of 34.3±11.7 years, 73.1% were transmen and 26.9% transwomen and 88.9% were undergoing gender-affirming hormonal treatment (GAHT). One in four subjects (24.1%) presented a moderate-to-severe impact of the levitra event (Impact of Event Scale buy levitra vardenafil score ≥26). The availability of telematic endocrinological visit was associated with better Mental Health Scores in the buy levitra vardenafil 12-items Short Form Health Survey(SF-12) (p=0.030) and better IES (p=0.006).Our survey suggests a positive effect of telemedicine as the availability of telematic endocrinological consultations may have relieved the distress caused by the levitra by offering the opportunity to avoid halting GAHT.

In fact, deprivation of GAHT may result in several negative effects such as the increase buy levitra vardenafil in short-term self-medication and in depression and suicidal behaviour not only for those waiting for the start of treatment but also for those already using hormones.5 In conclusion, particular attention should be paid to vulnerable groups like the transpopulation who may pay a higher price during the levitra. The use of telemedicine for continuation and monitoring of GAHT may be an effective tool for mitigating the negative effects of the levitra.AcknowledgmentsThe authors thank Julie Norbury for English copy editing.The British Medical Association recently published their report on the impact of erectile dysfunction treatment on mental health in England, highlighting the urgent need for investment in mental health services and further recruitment of mental health staff.1 Like many others, they have predicted a substantial increase in demand buy levitra vardenafil on mental health services in the coming months. Their recommendations include a call for detailed workforce planning at local, national and system buy levitra vardenafil levels. This coincides with the publication of the ‘NHS People Plan’ which also emphasised the need to maximise staff potential.2 The message from both is clear, it is time for Trusts to revise and improve how they use their multidisciplinary workforce, including non-medical prescribers (NMPs).Pharmacists have been able to register as independent prescribers since 20063 and as such, can work autonomously to prescribe any medicine for any medical condition within their areas of competency.4 There has been a slow uptake of pharmacists into this role5 and while a recent General Pharmaceutical Council survey found only buy levitra vardenafil a small increase between the number of active prescribers from 2013 (1.094) to 2019 (1.590), almost a quarter of prescribers included mental health within their prescribing practice.6 More recently, we have started to see increasing reports of the value of pharmacist independent prescribers in mental health services.7 8Pharmacists bring a unique perspective to patient consultation.

Their expertise in pharmacology and medicine use means they are ideally placed to help patients optimise their medicines treatment4 and to ensure that patients are involved in decisions about their medicines, taking into account individual views and preferences. This approach is consistent with the guidance on medicines optimisation from the National Institute for Health and Care Excellence9 and the Royal Pharmaceutical Society,10 and the Department of Health’s drive to involve patients actively in clinical decisions.11 An increased focus on precision psychiatry in urging clinicians to tailor medicines to patients according to evidence about individualised risks and benefits.12 13 However, it takes buy levitra vardenafil time to discuss medicine choices and to explore individual beliefs about medicines. This is especially relevant buy levitra vardenafil in Psychiatry, where a large group of medicines (eg, antipsychotics) may have a wide range of potential side effects. Prescribing pharmacists could provide leadership and support in tailoring medicines for patients, as part of the wider multidisciplinary team.10The recent news that Priadel, the most commonly used brand of lithium in the UK, is planned to be discontinued14 is buy levitra vardenafil another example where a new and unexpected burden on psychiatric services could be eased by sharing the workload with prescribing pharmacists.

The Medicines and Healthcare Products Regulatory Agency recommends that patients should have an individualised medication review in order to switch from one brand of lithium to another.14 This is work that can be done by prescribing pharmacists who have an in-depth knowledge of the pharmacokinetics of lithium formulations.Importantly, this is a role that can buy levitra vardenafil be delivered using telepsychiatry and enhanced by the use of digital tools. Patients can meet pharmacists from buy levitra vardenafil the comfort of their own home using video conferencing. Pharmacists can upload and share medicines information on the screen while discussing the benefits, risks and individual medication needs with each client. Increasingly organisations are using technology whereby prescriptions can be prepared electronically and sent securely to patients or their medicines providers.15We know from systematic reviews that NMPs in general are considered to provide a responsive, efficient and convenient service5 and to deliver similar prescribing outcomes as doctors.16 Medical professionals who have worked with NMPs have found that this support permits them to concentrate on clinical issues that require medical expertise.5 A patient survey buy levitra vardenafil carried out in 2013 indicated that independent non‐medical prescribing was valued highly by patients and that generally there were few perceived differences in the care received from respondents’ NMP and their usual doctor.17 The literature also suggests that an NMP’s role is more likely to flourish when linked to a strategic vision of NMPs within an National Health Service (NHS) Trust, along with a well-defined area of practice.18Mental health trusts are being asked to prepare for a surge in referrals and as part of this planning, they will need to ensure that they get the most out of their highly skilled workforce.

There are active pharmacist prescribers in many trusts, however, this role is not yet commonplace.19 Health Education England has already identified that this is an important area of transformation for pharmacy and has called on mental health pharmacy teams to develop and share innovative ways of working.19 The ‘NHS People Plan’ outlines a commitment to train 50 community-based specialist mental health pharmacists within the next 2 years, along with a plan to extend the pharmacy foundation training to create a sustainable supply of prescribing pharmacists in future years.2We suggest that Mental Health Trusts should urgently develop prescribing roles for specialist mental health pharmacists, which are integrated buy levitra vardenafil within mental health teams. In these roles, prescribing pharmacists can actively support their multidisciplinary buy levitra vardenafil colleagues in case discussion meetings. Furthermore, they should buy levitra vardenafil host regular medication review clinics, where patients can be referred to discuss their medicine options and, as advancements in precision therapeutics continue, have their treatment individually tailored to their needs. This is the way forward for a modern and patient-oriented NHS in the UK..

The transpopulation represents a vulnerable population segment both socially and how to get a levitra prescription from your doctor medically, how to get levitra online with a higher incidence of mental health issues. During the erectile dysfunction treatment outbreak, transgender persons have faced additional social, psychological how to get levitra online and physical difficulties.1 2 In Italy and in several other countries access to healthcare has been difficult or impossible thereby hindering the start or continuation of hormonal and psychological treatments. Furthermore, several planned gender-affirming surgeries have been how to get levitra online postponed. These obstacles may have caused an additional psychological burden given the positive effects of medical and surgical treatments on well-being, how to get levitra online directly and indirectly, reducing stressors such as workplace discrimination and social inequalities.3 Some organisational aspects should also be considered.

Binary gender policies may worsen inequalities and marginalisation of transgender subjects potentially increasing the risk of morbidity and mortality.As with the general population, during the lockdown, the Internet and social media were useful in reducing isolation and, in this particular population, were also relevant how to get levitra online for keeping in touch with associations and healthcare facilities with the support of telemedicine services.4 Addressing the role of the telemedicine in the transpopulation, between May and June 2020 we conducted an anonymous web-based survey among transgenders living in Italy (ClinicalTrials.gov Identifier NCT04448418). Among the 108 respondents, with a mean age of 34.3±11.7 years, 73.1% were transmen and 26.9% transwomen and 88.9% were undergoing gender-affirming hormonal treatment (GAHT). One in four subjects (24.1%) presented a moderate-to-severe impact of the levitra how to get levitra online event (Impact of Event Scale score ≥26). The availability of telematic endocrinological visit was associated with better Mental how to get levitra online Health Scores in the 12-items Short Form Health Survey(SF-12) (p=0.030) and better IES (p=0.006).Our survey suggests a positive effect of telemedicine as the availability of telematic endocrinological consultations may have relieved the distress caused by the levitra by offering the opportunity to avoid halting GAHT.

In fact, deprivation of GAHT may result in several negative effects such as the increase in short-term self-medication and in depression and suicidal behaviour not only for those waiting for the start of treatment but also for those already using hormones.5 In conclusion, particular attention should be how to get levitra online paid to vulnerable groups like the transpopulation who may pay a higher price during the levitra. The use of how to get levitra online telemedicine for continuation and monitoring of GAHT may be an effective tool for mitigating the negative effects of the levitra.AcknowledgmentsThe authors thank Julie Norbury for English copy editing.The British Medical Association recently published their report on the impact of erectile dysfunction treatment on mental health in England, highlighting the urgent need for investment in mental health services and further recruitment of mental health staff.1 Like many others, they have predicted a substantial increase in demand on mental health services in the coming months. Their recommendations include a call for detailed workforce how to get levitra online planning at local, national and system levels. This coincides with the publication of the ‘NHS People Plan’ which also emphasised the need to maximise staff potential.2 The message from both is clear, it is time for Trusts to revise and improve how they use their multidisciplinary workforce, including non-medical prescribers (NMPs).Pharmacists have been able to register as independent prescribers since 20063 and as how to get levitra online such, can work autonomously to prescribe any medicine for any medical condition within their areas of competency.4 There has been a slow uptake of pharmacists into this role5 and while a recent General Pharmaceutical Council survey found only a small increase between the number of active prescribers from 2013 (1.094) to 2019 (1.590), almost a quarter of prescribers included mental health within their prescribing practice.6 More recently, we have started to see increasing reports of the value of pharmacist independent prescribers in mental health services.7 8Pharmacists bring a unique perspective to patient consultation.

Their expertise in pharmacology and medicine use means they are ideally placed to help patients optimise their medicines treatment4 and to ensure that patients are involved in decisions about their medicines, taking into account individual views and preferences. This approach is consistent with the guidance on medicines optimisation from the National Institute for Health and Care Excellence9 and the Royal Pharmaceutical Society,10 and the Department of Health’s drive to involve patients actively in clinical decisions.11 An increased focus on precision psychiatry in urging clinicians to tailor medicines to patients according to evidence about individualised risks and benefits.12 13 However, it takes time to discuss medicine choices and to explore how to get levitra online individual beliefs about medicines. This is especially relevant in Psychiatry, where a large group of medicines (eg, antipsychotics) may have how to get levitra online a wide range of potential side effects. Prescribing pharmacists could provide leadership and support in tailoring medicines for patients, as part of the wider multidisciplinary team.10The recent news that Priadel, the most commonly used brand of lithium in the how to get levitra online UK, is planned to be discontinued14 is another example where a new and unexpected burden on psychiatric services could be eased by sharing the workload with prescribing pharmacists.

The Medicines and Healthcare Products Regulatory Agency recommends that patients should have an individualised medication review in order to switch from one brand of lithium to another.14 This how to get levitra online is work that can be done by prescribing pharmacists who have an in-depth knowledge of the pharmacokinetics of lithium formulations.Importantly, this is a role that can be delivered using telepsychiatry and enhanced by the use of digital tools. Patients can meet pharmacists from the comfort of their own home using how to get levitra online video conferencing. Pharmacists can upload and share medicines information on the screen while discussing the benefits, risks and individual medication needs with each client. Increasingly organisations are using technology whereby prescriptions can be prepared electronically and sent securely to patients or their medicines providers.15We know from systematic reviews that NMPs in general are considered to provide a responsive, efficient and convenient service5 and to deliver similar prescribing outcomes how to get levitra online as doctors.16 Medical professionals who have worked with NMPs have found that this support permits them to concentrate on clinical issues that require medical expertise.5 A patient survey carried out in 2013 indicated that independent non‐medical prescribing was valued highly by patients and that generally there were few perceived differences in the care received from respondents’ NMP and their usual doctor.17 The literature also suggests that an NMP’s role is more likely to flourish when linked to a strategic vision of NMPs within an National Health Service (NHS) Trust, along with a well-defined area of practice.18Mental health trusts are being asked to prepare for a surge in referrals and as part of this planning, they will need to ensure that they get the most out of their highly skilled workforce.

There are active pharmacist prescribers in many trusts, however, this role is not yet commonplace.19 Health Education England has already identified that this is an important area of transformation for pharmacy and has called on mental health pharmacy teams to develop and share innovative ways of working.19 The ‘NHS People Plan’ outlines a commitment to train 50 community-based specialist mental health pharmacists within the next 2 years, along with a plan to extend the pharmacy foundation training to create a sustainable supply of prescribing pharmacists in future years.2We suggest that Mental Health Trusts should urgently how to get levitra online develop prescribing roles for specialist mental health pharmacists, which are integrated within mental health teams. In these roles, prescribing pharmacists can actively support their multidisciplinary colleagues in case discussion how to get levitra online meetings. Furthermore, they should host regular medication review clinics, where patients can be referred to discuss their medicine options and, as advancements in how to get levitra online precision therapeutics continue, have their treatment individually tailored to their needs. This is the way forward for a modern and patient-oriented NHS in the UK..

Levitra prospecto

IntroductionGLI-Kruppel family member 3 (GLI3) encodes for a zinc finger http://whiterockboatclub.com/home/ transcription factor which plays a key role in the sonic hedgehog (SHH) signalling pathway essential in both limb and craniofacial development.1 2 In hand development, SHH is levitra prospecto expressed in the zone of polarising activity (ZPA) on the posterior side of the handplate. The ZPA expresses SHH, creating a gradient of SHH from the posterior to the anterior side of the handplate. In the presence of SHH, full length GLI3-protein is produced (GLI3A), whereas absence of SHH causes cleavage of GLI3 into its levitra prospecto repressor form (GLI3R).3 4 Abnormal expression of this SHH/GLI3R gradient can cause both preaxial and postaxial polydactyly.2Concordantly, pathogenic DNA variants in the GLI3 gene are known to cause multiple syndromes with craniofacial and limb involvement, such as. Acrocallosal syndrome5 (OMIM.

200990), Greig levitra prospecto cephalopolysyndactyly syndrome6 (OMIM. 175700) and Pallister-Hall syndrome7 (OMIM. 146510). Also, in non-syndromic polydactyly, such as preaxial polydactyly-type 4 (PPD4, OMIM.

174700),8 pathogenic variants in GLI3 have been described. Out of these diseases, Pallister-Hall syndrome is the most distinct entity, defined by the presence of central polydactyly and hypothalamic hamartoma.9 The other GLI3 syndromes are defined by the presence of preaxial and/or postaxial polydactyly of the hand and feet with or without syndactyly (Greig syndrome, PPD4). Also, various mild craniofacial features such as hypertelorism and macrocephaly can occur. Pallister-Hall syndrome is caused by truncating variants in the middle third of the GLI3 gene.10–12 The truncation of GLI3 causes an overexpression of GLI3R, which is believed to be the key difference between Pallister-Hall and the GLI3-mediated polydactyly syndromes.9 11 Although multiple attempts have been made, the clinical and genetic distinction between the GLI3-mediated polydactyly syndromes is less evident.

This has for example led to the introduction of subGreig and the formulation of an Oro-facial-digital overlap syndrome.10 Other authors, suggested that we should not regard these diseases as separate entities, but as a spectrum of GLI3-mediated polydactyly syndromes.13Although phenotype/genotype correlation of the different syndromes has been cumbersome, clinical and animal studies do provide evidence that distinct regions within the gene, could be related to the individual anomalies contributing to these syndromes. First, case studies show isolated preaxial polydactyly is caused by both truncating and non-truncating variants throughout the GLI3 gene, whereas in isolated postaxial polydactyly cases truncating variants at the C-terminal side of the gene are observed.12 14 These results suggest two different groups of variants for preaxial and postaxial polydactyly. Second, recent animal studies suggest that posterior malformations in GLI3-mediated polydactyly syndromes are likely related to a dosage effect of GLI3R rather than due to the influence of an altered GLI3A expression.15Past attempts for phenotype/genotype correlation in GLI3-mediated polydactyly syndromes have directly related the diagnosed syndrome to the observed genotype.10–12 16 Focusing on individual hand phenotypes, such as preaxial and postaxial polydactyly and syndactyly might be more reliable because it prevents misclassification due to inconsistent use of syndrome definition. Subsequently, latent class analysis (LCA) provides the possibility to relate a group of observed variables to a set of latent, or unmeasured, parameters and thereby identifying different subgroups in the obtained dataset.17 As a result, LCA allows us to group different phenotypes within the GLI3-mediated polydactyly syndromes and relate the most important predictors of the grouped phenotypes to the observed GLI3 variants.The aim of our study was to further investigate the correlation of the individual phenotypes to the genotypes observed in GLI3-mediated polydactyly syndromes, using LCA.

Cases were obtained by both literature review and the inclusion of local clinical cases. Subsequently, we identified two subclasses of limb anomalies that relate to the underlying GLI3 variant. We provide evidence for two different phenotypic and genotypic groups with predominantly preaxial and postaxial hand and feet anomalies, and we specify those cases with a higher risk for corpus callosum anomalies.MethodsLiterature reviewThe Human Gene Mutation Database (HGMD Professional 2019) was reviewed to identify known pathogenic variants in GLI3 and corresponding phenotypes.18 All references were obtained and cases were included when they were diagnosed with either Greig or subGreig syndrome or PPD4.10–12 Pallister-Hall syndrome and acrocallosal syndrome were excluded because both are regarded distinct syndromes and rather defined by the presence of the non-hand anomalies, than the presence of preaxial or postaxial polydactyly.13 19 Isolated preaxial or postaxial polydactyly were excluded for two reasons. The phenotype/genotype correlations are better understood and both anomalies can occur sporadically which could introduce falsely assumed pathogenic GLI3 variants in the analysis.

Additionally, cases were excluded when case-specific phenotypic or genotypic information was not reported or if these two could not be related to each other. Families with a combined phenotypic description, not reducible to individual family members, were included as one case in the analysis.Clinical casesThe Sophia Children’s Hospital Database was reviewed for cases with a GLI3 variant. Within this population, the same inclusion criteria for the phenotype were valid. Relatives of the index patients were also contacted for participation in this study, when they showed comparable hand, foot, or craniofacial malformations or when a GLI3 variant was identified.

Phenotypes of the hand, foot and craniofacial anomalies of the patients treated in the Sophia Children's Hospital were collected using patient documentation. Family members were identified and if possible, clinically verified. Alternatively, family members were contacted to verify their phenotypes. If no verification was possible, cases were excluded.PhenotypesThe phenotypes of both literature cases and local cases were extracted in a similar fashion.

The most frequently reported limb and craniofacial phenotypes were dichotomised. The dichotomised hand and foot phenotypes were preaxial polydactyly, postaxial polydactyly and syndactyly. Broad halluces or thumbs were commonly reported by authors and were dichotomised as a presentation of preaxial polydactyly. The extracted dichotomised craniofacial phenotypes were hypertelorism, macrocephaly and corpus callosum agenesis.

All other phenotypes were registered, but not dichotomised.Pathogenic GLI3 variantsAll GLI3 variants were extracted and checked using Alamut Visual V.2.14. If indicated, variants were renamed according to standard Human Genome Variation Society nomenclature.20 Variants were grouped in either missense, frameshift, nonsense or splice site variants. In the group of frameshift variants, a subgroup with possible splice site effect were identified for subgroup analysis when indicated. Similarly, nonsense variants prone for nonsense mediated decay (NMD) and nonsense variants with experimentally confirmed NMD were identified.21 Deletions of multiple exons, CNVs and translocations were excluded for analysis.

A full list of included mutations is available in the online supplementary materials.Supplemental materialThe location of the variant was compared with five known structural domains of the GLI3 gene. (1) repressor domain, (2) zinc finger domain, (3) cleavage site, (4) activator domain, which we defined as a concatenation of the separately identified transactivation zones, the CBP binding domain and the mediator binding domain (MBD) and (5) the MID1 interaction region domain.1 6 22–24 The boundaries of each of the domains were based on available literature (figure 1, exact locations available in the online supplementary materials). The boundaries used by different authors did vary, therefore a consensus was made.In this figure the posterior probability of an anterior phenotype is plotted against the location of the variant, stratified for the type of mutation that was observed. For better overview, only variants with a location effect were displayed.

The full figure, including all variant types, can be found in the online supplementary figure 1. Each mutation is depicted as a dot, the size of the dot represents the number of observations for that variant. If multiple observations were made, the mean posterior odds and IQR are plotted. For the nonsense variants, variants that were predicted to produce nonsense mediated decay, are depicted using a triangle.

Again, the size indicates the number of observations." data-icon-position data-hide-link-title="0">Figure 1 In this figure the posterior probability of an anterior phenotype is plotted against the location of the variant, stratified for the type of mutation that was observed. For better overview, only variants with a location effect were displayed. The full figure, including all variant types, can be found in the online supplementary figure 1. Each mutation is depicted as a dot, the size of the dot represents the number of observations for that variant.

If multiple observations were made, the mean posterior odds and IQR are plotted. For the nonsense variants, variants that were predicted to produce nonsense mediated decay, are depicted using a triangle. Again, the size indicates the number of observations.Supplemental materialLatent class analysisTo cluster phenotypes and relate those to the genotypes of the patients, an explorative analysis was done using LCA in R (R V.3.6.1 for Mac. Polytomous variable LCA, poLCA V.1.4.1.).

We used our LCA to detect the number of phenotypic subgroups in the dataset and subsequently predict a class membership for each case in the dataset based on the posterior probabilities.In order to make a reliable prediction, only phenotypes that were sufficiently reported and/or ruled out were feasible for LCA, limiting the analysis to preaxial polydactyly, postaxial polydactyly and syndactyly of the hands and feet. Only full cases were included. To determine the optimal number of classes, we fitted a series of models ranging from a one-class to a six-class model. The optimal number of classes was based on the conditional Akaike information criterion (cAIC), the non adjusted and the sample-size adjusted Bayesian information criterion (BIC and aBIC) and the obtained entropy.25 The explorative LCA produces both posterior probabilities per case for both classes and predicted class membership.

Using the predicted class membership, the phenotypic features per class were determined in a univariate analysis (χ2, SPSS V.25). Using the posterior probabilities on latent class (LC) membership, a scatter plot was created using the location of the variant on the x-axis and the probability of class membership on the y-axis for each of the types of variants (Tibco Spotfire V.7.14). Using these scatter plots, variants that give similar phenotypes were clustered.Genotype/phenotype correlationBecause an LC has no clinical value, the correlation between genotypes and phenotypes was investigated using the predictor phenotypes and the clustered phenotypes. First, those phenotypes that contribute most to LC membership were identified.

Second those phenotypes were directly related to the different types of variants (missense, nonsense, frameshift, splice site) and their clustered locations. Quantification of the relation was performed using a univariate analysis using a χ2 test. Because of our selection criteria, meaning patients at least have two phenotypes, a multivariate using a logistic regression analysis was used to detect the most significant predictors in the overall phenotype (SPSS V.25). Finally, we explored the relation of the clustered genotypes to the presence of corpus callosum agenesis, a rare malformation in GLI3-mediated polydactyly syndromes which cannot be readily diagnosed without additional imaging.ResultsWe included 251 patients from the literature and 46 local patients,10–12 16 21 26–43 in total 297 patients from 155 different families with 127 different GLI3 variants, 32 of which were large deletions, CNVs or translocations.

In six local cases, the exact variant could not be retrieved by status research.The distribution of the most frequently observed phenotypes and variants are presented in table 1. Other recurring phenotypes included developmental delay (n=22), broad nasal root (n=23), frontal bossing or prominent forehead (n=16) and craniosynostosis (n=13), camptodactyly (n=8) and a broad first interdigital webspace of the foot (n=6).View this table:Table 1 Baseline phenotypes and genotypes of selected populationThe LCA model was fitted using the six defined hand/foot phenotypes. Model fit indices for the LCA are displayed in table 2. Based on the BIC, a two-class model has the best fit for our data.

The four-class model does show a gain in entropy, however with a higher BIC and loss of df. Therefore, based on the majority of performance statistics and the interpretability of the model, a two-class model was chosen. Table 3 displays the distribution of phenotypes and genotypes over the two classes.View this table:Table 2 Model fit indices for the one-class through six-class model evaluated in our LCAView this table:Table 3 Distribution of phenotypes and genotypes in the two latent classes (LC)Table 1 depicts the baseline phenotypes and genotypes in the obtained population. Note incomplete data especially in the cranium phenotypes.

In total 259 valid genotypes were present. In total, 289 cases had complete data for all hand and foot address phenotypes (preaxial polydactyly, postaxial polydactyly and syndactyly) and thus were available for LCA. Combined, for phenotype/genotype correlation 258 cases were available with complete genotypes and complete hand and foot phenotypes.Table 2 depicts the model fit indices for all models that have been fitted to our data.Table 3 depicts the distribution of phenotypes and genotypes over the two assigned LCs. Hand and foot phenotypes were used as input for the LCA, thus are all complete cases.

Malformation of the cranium and genotypes do have missing cases. Note that for the LCA, full case description was required, resulting in eight cases due to incomplete phenotypes. Out of these eight, one also had a genotype that thus needed to be excluded. Missingness of genotypic data was higher in LC2, mostly due to CNVs (table 1).In 54/60 cases, a missense variant produced a posterior phenotype.

Likewise, splice site variants show the same phenotype in 23/24 cases (table 3). For both frameshift and nonsense variants, this relation is not significant (52 anterior vs 54 posterior and 26 anterior vs 42 posterior, respectively). Therefore, only for nonsense and frameshift variants the location of the variant was plotted against the probability for LC2 membership in figure 1. A full scatterplot of all variants is available in online supplementary figure 1.Figure 1 reveals a pattern for these nonsense and frameshift variants that reveals that variants at the C-terminal of the gene predict anterior phenotypes.

When relating the domains of the GLI3 protein to the observed phenotype, we observe that the majority of patients with a nonsense or frameshift variant in the repressor domain, the zinc finger domain or the cleavage site had a high probability of an LC2/anterior phenotype. This group contains all variants that are either experimentally determined to be subject to NMD (triangle marker in figure 1) or predicted to be subject to NMD (diamond marker in figure 1). Frameshift and nonsense variants in the activator domain result in high probability for an LC1/posterior phenotype. These variants will be further referred to as truncating variants in the activator domain.The univariate relation of the individual phenotypes to these two groups of variants are estimated and presented in table 4.

In our multivariate analysis, postaxial polydactyly of the foot and hand are the strongest predictors (Beta. 2.548, p<0001 and Beta. 1.47, p=0.013, respectively) for patients to have a truncating variant in the activator domain. Moreover, the effect sizes of preaxial polydactyly of the hand and feet (Beta.

ˆ’0.797, p=0123 and −1.772, p=0.001) reveals that especially postaxial polydactyly of the foot is the dominant predictor for the genetic substrate of the observed anomalies.View this table:Table 4 Univariate and multivariate analysis of the phenotype/genotype correlationTable 4 shows exploration of the individual phenotypes on the genotype, both univariate and multivariate. The multivariate analysis corrects for the presence of multiple phenotypes in the underlying population.Although the craniofacial anomalies could not be included in the LCA, the relation between the observed anomalies and the identified genetic substrates can be studied. The prevalence of hypertelorism was equally distributed over the two groups of variants (47/135 vs 21/47 respectively, p<0.229). However for corpus callosum agenesis and macrocephaly, there was a higher prevalence in patients with a truncating variant in the activator domain (3/75 vs 11/41, p<0.001.

OR. 8.8, p<0.001) and 42/123 vs 24/48, p<0.05). Noteworthy is the fact that 11/14 cases with corpus callosum agenesis in the dataset had a truncating variant in the activator domain.DiscussionIn this report, we present new insights into the correlation between the phenotype and the genotype in patients with GLI3-mediated polydactyly syndromes. We illustrate that there are two LCs of patients, best predicted by postaxial polydactyly of the hand and foot for LC1, and the preaxial polydactyly of the hand and foot and syndactyly of the foot for LC2.

Patients with postaxial phenotypes have a higher risk of having a truncating variant in the activator domain of the GLI3 gene which is also related to a higher risk of corpus callosum agenesis. These results suggest a functional difference between truncating variants on the N-terminal and the C-terminal side of the GLI3 cleavage site.Previous attempts of phenotype to genotype correlation have not yet provided the clinical confirmation of these assumed mechanisms in the pathophysiology of GLI3-mediated polydactyly syndromes. Johnston et al have successfully determined the Pallister-Hall region in which truncating variants produce a Pallister-Hall phenotype rather than Greig syndrome.11 However, in their latest population study, subtypes of both syndromes were included to explain the full spectrum of observed malformations. In 2015, Demurger et al reported the higher incidence of corpus callosum agenesis in the Greig syndrome population with truncating mutations in the activator domain.12 Al-Qattan in his review summarises the concept of a spectrum of anomalies dependent on haplo-insufficiency (through different mechanisms) and repressor overexpression.13 However, he bases this theory mainly on reviewed experimental data.

Our report is the first to provide an extensive clinical review of cases that substantiate the phenotypic difference between the two groups that could fit the suggested mechanisms. We agree with Al-Qattan et al that a variation of anomalies can be observed given any pathogenic variant in the GLI3 gene, but overall two dominant phenotypes are present. A population with predominantly preaxial anomalies and one with postaxial anomalies. The presence of preaxial or postaxial polydactyly and syndactyly is not mutually exclusive for one of these two subclasses.

Meaning that preaxial polydactyly can co-occur with postaxial polydactyly. However, truncating mutations in the activator domain produce a postaxial phenotype, as can be derived from the risk in table 4. The higher risk of corpus callosum agenesis in this population shows that differentiating between a preaxial phenotype and a postaxial phenotype, instead of between the different GLI3-mediated polydactyly syndromes, might be more relevant regarding diagnostics for corpus callosum agenesis.We chose to use LCA as an exploratory tool only in our population for two reasons. First of all, LCA can be useful to identify subgroups, but there is no ‘true’ model or number of subgroups you can detect.

The best fitting model can only be estimated based on the available measures and approximates the true subgroups that might be present. Second, LC membership assignment is a statistical procedure based on the posterior probability, with concordant errors of the estimation, rather than a clinical value that can be measured or evaluated. Therefore, we decided to use our LCA only in an exploratory tool, and perform our statistics using the actual phenotypes that predict LC membership and the associated genotypes. Overall, this method worked well to differentiate the two subgroups present in our dataset.

However, outliers were observed. A qualitative analysis of these outliers is available in the online supplementary data.The genetic substrate for the two phenotypic clusters can be discussed based on multiple experiments. Overall, we hypothesise two genetic clusters. One that is due to haploinsufficiency and one that is due to abnormal truncation of the activator.

The hypothesised cluster of variants that produce haploinsufficiency is mainly based on the experimental data that confirms NMD in two variants and the NMD prediction of other nonsense variants in Alamut. For the frameshift variants, it is also likely that the cleavage of the zinc finger domain results in functional haploinsufficiency either because of a lack of signalling domains or similarly due to NMD. Missense variants could cause haploinsufficiency through the suggested mechanism by Krauss et al who have illustrated that missense variants in the MID1 domain hamper the functional interaction with the MID1-α4-PP2A complex, leading to a subcellular location of GLI3.24 The observed missense variants in our study exceed the region to which Krauss et al have limited the MID-1 interaction domain. An alternative theory is suggested by Zhou et al who have shown that missense variants in the MBD can cause deficiency in the signalling of GLI3A, functionally implicating a relative overexpression of GLI3R.22 However, GLI3R overexpression would likely produce a posterior phenotype, as determined by Hill et al in their fixed homo and hemizygous GLI3R models.15 Therefore, our hypothesis is that all included missense variants have a similar pathogenesis which is more likely in concordance with the mechanism introduced by Krauss et al.

To our knowledge, no splice site variants have been functionally described in literature. However, it is noted that the 15 and last exon encompasses the entire activator domain, thus any splice site mutation is by definition located on the 5′ side of the activator. Based on the phenotype, we would suggest that these variants fail to produce a functional protein. We hypothesise that the truncating variants of the activator domain lead to overexpression of GLI3R in SHH rich areas.

In normal development, the presence of SHH prevents the processing of full length GLI34 into GLI3R, thus producing the full length activator. In patients with a truncating variant of the activator domain of GLI3, thus these variants likely have the largest effect in SHH rich areas, such as the ZPA located at the posterior side of the hand/footplate. Moreover, the lack of posterior anomalies in the GLI3∆699/- mouse model (hemizygous fixed repressor model) compared with the GLI3∆699/∆699 mouse model (homozygous fixed repressor model), suggesting a dosage effect of GLI3R to be responsible for posterior hand anomalies.15 These findings are supported by Lewandowski et al, who show that the majority of the target genes in GLI signalling are regulated by GLI3R rather than GLI3A.44 Together, these findings suggest a role for the location and type of variant in GLI3-mediated syndromes.Interestingly, the difference between Pallister-Hall syndrome and GLI3-mediated polydactyly syndromes has also been attributed to the GLI3R overexpression. However, the difference in phenotype observed in the cases with a truncating variant in the activator domain and Pallister-Hall syndrome suggest different functional consequences.

When studying figure 1, it is noted that the included truncating variants on the 3′ side of the cleavage site seldomly affect the CBP binding region, which could provide an explanation for the observed differences. This binding region is included in the Pallister-Hall region as defined by Johnston et al and is necessary for the downstream signalling with GLI1.10 11 23 45 Interestingly, recent reports show that pathogenic variants in GLI1 can produce phenotypes concordant with Ellis von Krefeld syndrome, which includes overlapping features with Pallister-Hall syndrome.46 The four truncating variants observed in this study that do affect the CBP but did not result in a Pallister-Hall phenotype are conflicting with this theory. Krauss et al postulate an alternative hypothesis, they state that the MID1-α4-PP2A complex, which is essential for GLI3A signalling, could also be the reason for overlapping features of Opitz syndrome, caused by variants in MID1, and Pallister-Hall syndrome. Further analysis is required to fully appreciate the functional differences between truncating mutations that cause Pallister-Hall syndrome and those that result in GLI3-mediated polydactyly syndromes.For the clinical evaluation of patients with GLI3-mediated polydactyly syndromes, intracranial anomalies are likely the most important to predict based on the variant.

Unfortunately, the presence of corpus callosum agenesis was not routinely investigated or reported thus this feature could not be used as an indicator phenotype for LC membership. Interestingly when using only hand and foot phenotypes, we did notice a higher prevalence of corpus callosum agenesis in patients with posterior phenotypes. The suggested relation between truncating mutations in the activator domain causing these posterior phenotypes and corpus callosum agenesis was statistically confirmed (OR. 8.8, p<0.001).

Functionally this relation could be caused by the GLI3-MED12 interaction at the MBD. Pathogenic DNA variants in MED12 can cause Opitz-Kaveggia syndrome, a syndrome in which presentation includes corpus callosum agenesis, broad halluces and thumbs.47In conclusion, there are two distinct phenotypes within the GLI3-mediated polydactyly population. Patients with more posteriorly and more anteriorly oriented hand anomalies. Furthermore, this difference is related to the observed variant in GLI3.

We hypothesise that variants that cause haploinsufficiency produce anterior anomalies of the hand, whereas variants with abnormal truncation of the activator domain have more posterior anomalies. Furthermore, patients that have a variant that produces abnormal truncation of the activator domain, have a greater risk for corpus callosum agenesis. Thus, we advocate to differentiate preaxial or postaxial oriented GLI3 phenotypes to explain the pathophysiology as well as to get a risk assessment for corpus callosum agenesis.Data availability statementData are available upon reasonable request.Ethics statementsPatient consent for publicationNot required.Ethics approvalThe research protocol was approved by the local ethics board of the Erasmus MC University Medical Center (MEC 2015-679)..

IntroductionGLI-Kruppel family member 3 (GLI3) encodes for a zinc finger transcription factor which plays a key role in the sonic hedgehog (SHH) signalling pathway essential in both limb and craniofacial development.1 2 In hand development, how to get levitra online SHH is expressed in the zone of polarising activity (ZPA) on the image source posterior side of the handplate. The ZPA expresses SHH, creating a gradient of SHH from the posterior to the anterior side of the handplate. In the presence of SHH, full length GLI3-protein is produced (GLI3A), whereas absence of SHH causes cleavage of GLI3 how to get levitra online into its repressor form (GLI3R).3 4 Abnormal expression of this SHH/GLI3R gradient can cause both preaxial and postaxial polydactyly.2Concordantly, pathogenic DNA variants in the GLI3 gene are known to cause multiple syndromes with craniofacial and limb involvement, such as. Acrocallosal syndrome5 (OMIM. 200990), Greig cephalopolysyndactyly how to get levitra online syndrome6 (OMIM.

175700) and Pallister-Hall syndrome7 (OMIM. 146510). Also, in non-syndromic polydactyly, such as preaxial polydactyly-type 4 (PPD4, OMIM. 174700),8 pathogenic variants in GLI3 have been described. Out of these diseases, Pallister-Hall syndrome is the most distinct entity, defined by the presence of central polydactyly and hypothalamic hamartoma.9 The other GLI3 syndromes are defined by the presence of preaxial and/or postaxial polydactyly of the hand and feet with or without syndactyly (Greig syndrome, PPD4).

Also, various mild craniofacial features such as hypertelorism and macrocephaly can occur. Pallister-Hall syndrome is caused by truncating variants in the middle third of the GLI3 gene.10–12 The truncation of GLI3 causes an overexpression of GLI3R, which is believed to be the key difference between Pallister-Hall and the GLI3-mediated polydactyly syndromes.9 11 Although multiple attempts have been made, the clinical and genetic distinction between the GLI3-mediated polydactyly syndromes is less evident. This has for example led to the introduction of subGreig and the formulation of an Oro-facial-digital overlap syndrome.10 Other authors, suggested that we should not regard these diseases as separate entities, but as a spectrum of GLI3-mediated polydactyly syndromes.13Although phenotype/genotype correlation of the different syndromes has been cumbersome, clinical and animal studies do provide evidence that distinct regions within the gene, could be related to the individual anomalies contributing to these syndromes. First, case studies show isolated preaxial polydactyly is caused by both truncating and non-truncating variants throughout the GLI3 gene, whereas in isolated postaxial polydactyly cases truncating variants at the C-terminal side of the gene are observed.12 14 These results suggest two different groups of variants for preaxial and postaxial polydactyly. Second, recent animal studies suggest that posterior malformations in GLI3-mediated polydactyly syndromes are likely related to a dosage effect of GLI3R rather than due to the influence of an altered GLI3A expression.15Past attempts for phenotype/genotype correlation in GLI3-mediated polydactyly syndromes have directly related the diagnosed syndrome to the observed genotype.10–12 16 Focusing on individual hand phenotypes, such as preaxial and postaxial polydactyly and syndactyly might be more reliable because it prevents misclassification due to inconsistent use of syndrome definition.

Subsequently, latent class analysis (LCA) provides the possibility to relate a group of observed variables to a set of latent, or unmeasured, parameters and thereby identifying different subgroups in the obtained dataset.17 As a result, LCA allows us to group different phenotypes within the GLI3-mediated polydactyly syndromes and relate the most important predictors of the grouped phenotypes to the observed GLI3 variants.The aim of our study was to further investigate the correlation of the individual phenotypes to the genotypes observed in GLI3-mediated polydactyly syndromes, using LCA. Cases were obtained by both literature review and the inclusion of local clinical cases. Subsequently, we identified two subclasses of limb anomalies that relate to the underlying GLI3 variant. We provide evidence for two different phenotypic and genotypic groups with predominantly preaxial and postaxial hand and feet anomalies, and we specify those cases with a higher risk for corpus callosum anomalies.MethodsLiterature reviewThe Human Gene Mutation Database (HGMD Professional 2019) was reviewed to identify known pathogenic variants in GLI3 and corresponding phenotypes.18 All references were obtained and cases were included when they were diagnosed with either Greig or subGreig syndrome or PPD4.10–12 Pallister-Hall syndrome and acrocallosal syndrome were excluded because both are regarded distinct syndromes and rather defined by the presence of the non-hand anomalies, than the presence of preaxial or postaxial polydactyly.13 19 Isolated preaxial or postaxial polydactyly were excluded for two reasons. The phenotype/genotype correlations are better understood and both anomalies can occur sporadically which could introduce falsely assumed pathogenic GLI3 variants in the analysis.

Additionally, cases were excluded when case-specific phenotypic or genotypic information was not reported or if these two could not be related to each other. Families with a combined phenotypic description, not reducible to individual family members, were included as one case in the analysis.Clinical casesThe Sophia Children’s Hospital Database was reviewed for cases with a GLI3 variant. Within this population, the same inclusion criteria for the phenotype were valid. Relatives of the index patients were also contacted for participation in this study, when they showed comparable hand, foot, or craniofacial malformations or when a GLI3 variant was identified. Phenotypes of the hand, foot and craniofacial anomalies of the patients treated in the Sophia Children's Hospital were collected using patient documentation.

Family members were identified and if possible, clinically verified. Alternatively, family members were contacted to verify their phenotypes. If no verification was possible, cases were excluded.PhenotypesThe phenotypes of both literature cases and local cases were extracted in a similar fashion. The most frequently reported limb and craniofacial phenotypes were dichotomised. The dichotomised hand and foot phenotypes were preaxial polydactyly, postaxial polydactyly and syndactyly.

Broad halluces or thumbs were commonly reported by authors and were dichotomised as a presentation of preaxial polydactyly. The extracted dichotomised craniofacial phenotypes were hypertelorism, macrocephaly and corpus callosum agenesis. All other phenotypes were registered, but not dichotomised.Pathogenic GLI3 variantsAll GLI3 variants were extracted and checked using Alamut Visual V.2.14. If indicated, variants were renamed according to standard Human Genome Variation Society nomenclature.20 Variants were grouped in either missense, frameshift, nonsense or splice site variants. In the group of frameshift variants, a subgroup with possible splice site effect were identified for subgroup analysis when indicated.

Similarly, nonsense variants prone for nonsense mediated decay (NMD) and nonsense variants with experimentally confirmed NMD were identified.21 Deletions of multiple exons, CNVs and translocations were excluded for analysis. A full list of included mutations is available in the online supplementary materials.Supplemental materialThe location of the variant was compared with five known structural domains of the GLI3 gene. (1) repressor domain, (2) zinc finger domain, (3) cleavage site, (4) activator domain, which we defined as a concatenation of the separately identified transactivation zones, the CBP binding domain and the mediator binding domain (MBD) and (5) the MID1 interaction region domain.1 6 22–24 The boundaries of each of the domains were based on available literature (figure 1, exact locations available in the online supplementary materials). The boundaries used by different authors did vary, therefore a consensus was made.In this figure the posterior probability of an anterior phenotype is plotted against the location of the variant, stratified for the type of mutation that was observed. For better overview, only variants with a location effect were displayed.

The full figure, including all variant types, can be found in the online supplementary figure 1. Each mutation is depicted as a dot, the size of the dot represents the number of observations for that variant. If multiple observations were made, the mean posterior odds and IQR are plotted. For the nonsense variants, variants that were predicted to produce nonsense mediated decay, are depicted using a triangle. Again, the size indicates the number of observations." data-icon-position data-hide-link-title="0">Figure 1 In this figure the posterior probability of an anterior phenotype is plotted against the location of the variant, stratified for the type of mutation that was observed.

For better overview, only variants with a location effect were displayed. The full figure, including all variant types, can be found in the online supplementary figure 1. Each mutation is depicted as a dot, the size of the dot represents the number of observations for that variant. If multiple observations were made, the mean posterior odds and IQR are plotted. For the nonsense variants, variants that were predicted to produce nonsense mediated decay, are depicted using a triangle.

Again, the size indicates the number of observations.Supplemental materialLatent class analysisTo cluster phenotypes and relate those to the genotypes of the patients, an explorative analysis was done using LCA in R (R V.3.6.1 for Mac. Polytomous variable LCA, poLCA V.1.4.1.). We used our LCA to detect the number of phenotypic subgroups in the dataset and subsequently predict a class membership for each case in the dataset based on the posterior probabilities.In order to make a reliable prediction, only phenotypes that were sufficiently reported and/or ruled out were feasible for LCA, limiting the analysis to preaxial polydactyly, postaxial polydactyly and syndactyly of the hands and feet. Only full cases were included. To determine the optimal number of classes, we fitted a series of models ranging from a one-class to a six-class model.

The optimal number of classes was based on the conditional Akaike information criterion (cAIC), the non adjusted and the sample-size adjusted Bayesian information criterion (BIC and aBIC) and the obtained entropy.25 The explorative LCA produces both posterior probabilities per case for both classes and predicted class membership. Using the predicted class membership, the phenotypic features per class were determined in a univariate analysis (χ2, SPSS V.25). Using the posterior probabilities on latent class (LC) membership, a scatter plot was created using the location of the variant on the x-axis and the probability of class membership on the y-axis for each of the types of variants (Tibco Spotfire V.7.14). Using these scatter plots, variants that give similar phenotypes were clustered.Genotype/phenotype correlationBecause an LC has no clinical value, the correlation between genotypes and phenotypes was investigated using the predictor phenotypes and the clustered phenotypes. First, those phenotypes that contribute most to LC membership were identified.

Second those phenotypes were directly related to the different types of variants (missense, nonsense, frameshift, splice site) and their clustered locations. Quantification of the relation was performed using a univariate analysis using a χ2 test. Because of our selection criteria, meaning patients at least have two phenotypes, a multivariate using a logistic regression analysis was used to detect the most significant predictors in the overall phenotype (SPSS V.25). Finally, we explored the relation of the clustered genotypes to the presence of corpus callosum agenesis, a rare malformation in GLI3-mediated polydactyly syndromes which cannot be readily diagnosed without additional imaging.ResultsWe included 251 patients from the literature and 46 local patients,10–12 16 21 26–43 in total 297 patients from 155 different families with 127 different GLI3 variants, 32 of which were large deletions, CNVs or translocations. In six local cases, the exact variant could not be retrieved by status research.The distribution of the most frequently observed phenotypes and variants are presented in table 1.

Other recurring phenotypes included developmental delay (n=22), broad nasal root (n=23), frontal bossing or prominent forehead (n=16) and craniosynostosis (n=13), camptodactyly (n=8) and a broad first interdigital webspace of the foot (n=6).View this table:Table 1 Baseline phenotypes and genotypes of selected populationThe LCA model was fitted using the six defined hand/foot phenotypes. Model fit indices for the LCA are displayed in table 2. Based on the BIC, a two-class model has the best fit for our data. The four-class model does show a gain in entropy, however with a higher BIC and loss of df. Therefore, based on the majority of performance statistics and the interpretability of the model, a two-class model was chosen.

Table 3 displays the distribution of phenotypes and genotypes over the two classes.View this table:Table 2 Model fit indices for the one-class through six-class model evaluated in our LCAView this table:Table 3 Distribution of phenotypes and genotypes in the two latent classes (LC)Table 1 depicts the baseline phenotypes and genotypes in the obtained population. Note incomplete data especially in the cranium phenotypes. In total 259 valid genotypes were present. In total, 289 cases had complete data for all hand and foot phenotypes (preaxial polydactyly, postaxial polydactyly and syndactyly) and thus were available for LCA. Combined, for phenotype/genotype correlation 258 cases were available with complete genotypes and complete hand and foot phenotypes.Table 2 depicts the model fit indices for all models that have been fitted to our data.Table 3 depicts the distribution of phenotypes and genotypes over the two assigned LCs.

Hand and foot phenotypes were used as input for the LCA, thus are all complete cases. Malformation of the cranium and genotypes do have missing cases. Note that for the LCA, full case description was required, resulting in eight cases due to incomplete phenotypes. Out of these eight, one also had a genotype that thus needed to be excluded. Missingness of genotypic data was higher in LC2, mostly due to CNVs (table 1).In 54/60 cases, a missense variant produced a posterior phenotype.

Likewise, splice site variants show the same phenotype in 23/24 cases (table 3). For both frameshift and nonsense variants, this relation is not significant (52 anterior vs 54 posterior and 26 anterior vs 42 posterior, respectively). Therefore, only for nonsense and frameshift variants the location of the variant was plotted against the probability for LC2 membership in figure 1. A full scatterplot of all variants is available in online supplementary figure 1.Figure 1 reveals a pattern for these nonsense and frameshift variants that reveals that variants at the C-terminal of the gene predict anterior phenotypes. When relating the domains of the GLI3 protein to the observed phenotype, we observe that the majority of patients with a nonsense or frameshift variant in the repressor domain, the zinc finger domain or the cleavage site had a high probability of an LC2/anterior phenotype.

This group contains all variants that are either experimentally determined to be subject to NMD (triangle marker in figure 1) or predicted to be subject to NMD (diamond marker in figure 1). Frameshift and nonsense variants in the activator domain result in high probability for an LC1/posterior phenotype. These variants will be further referred to as truncating variants in the activator domain.The univariate relation of the individual phenotypes to these two groups of variants are estimated and presented in table 4. In our multivariate analysis, postaxial polydactyly of the foot and hand are the strongest predictors (Beta. 2.548, p<0001 and Beta.

1.47, p=0.013, respectively) for patients to have a truncating variant in the activator domain. Moreover, the effect sizes of preaxial polydactyly of the hand and feet (Beta. ˆ’0.797, p=0123 and −1.772, p=0.001) reveals that especially postaxial polydactyly of the foot is the dominant predictor for the genetic substrate of the observed anomalies.View this table:Table 4 Univariate and multivariate analysis of the phenotype/genotype correlationTable 4 shows exploration of the individual phenotypes on the genotype, both univariate and multivariate. The multivariate analysis corrects for the presence of multiple phenotypes in the underlying population.Although the craniofacial anomalies could not be included in the LCA, the relation between the observed anomalies and the identified genetic substrates can be studied. The prevalence of hypertelorism was equally distributed over the two groups of variants (47/135 vs 21/47 respectively, p<0.229).

However for corpus callosum agenesis and macrocephaly, there was a higher prevalence in patients with a truncating variant in the activator domain (3/75 vs 11/41, p<0.001. OR. 8.8, p<0.001) and 42/123 vs 24/48, p<0.05). Noteworthy is the fact that 11/14 cases with corpus callosum agenesis in the dataset had a truncating variant in the activator domain.DiscussionIn this report, we present new insights into the correlation between the phenotype and the genotype in patients with GLI3-mediated polydactyly syndromes. We illustrate that there are two LCs of patients, best predicted by postaxial polydactyly of the hand and foot for LC1, and the preaxial polydactyly of the hand and foot and syndactyly of the foot for LC2.

Patients with postaxial phenotypes have a higher risk of having a truncating variant in the activator domain of the GLI3 gene which is also related to a higher risk of corpus callosum agenesis. These results suggest a functional difference between truncating variants on the N-terminal and the C-terminal side of the GLI3 cleavage site.Previous attempts of phenotype to genotype correlation have not yet provided the clinical confirmation of these assumed mechanisms in the pathophysiology of GLI3-mediated polydactyly syndromes. Johnston et al have successfully determined the Pallister-Hall region in which truncating variants produce a Pallister-Hall phenotype rather than Greig syndrome.11 However, in their latest population study, subtypes of both syndromes were included to explain the full spectrum of observed malformations. In 2015, Demurger et al reported the higher incidence of corpus callosum agenesis in the Greig syndrome population with truncating mutations in the activator domain.12 Al-Qattan in his review summarises the concept of a spectrum of anomalies dependent on haplo-insufficiency (through different mechanisms) and repressor overexpression.13 However, he bases this theory mainly on reviewed experimental data. Our report is the first to provide an extensive clinical review of cases that substantiate the phenotypic difference between the two groups that could fit the suggested mechanisms.

We agree with Al-Qattan et al that a variation of anomalies can be observed given any pathogenic variant in the GLI3 gene, but overall two dominant phenotypes are present. A population with predominantly preaxial anomalies and one with postaxial anomalies. The presence of preaxial or postaxial polydactyly and syndactyly is not mutually exclusive for one of these two subclasses. Meaning that preaxial polydactyly can co-occur with postaxial polydactyly. However, truncating mutations in the activator domain produce a postaxial phenotype, as can be derived from the risk in table 4.

The higher risk of corpus callosum agenesis in this population shows that differentiating between a preaxial phenotype and a postaxial phenotype, instead of between the different GLI3-mediated polydactyly syndromes, might be more relevant regarding diagnostics for corpus callosum agenesis.We chose to use LCA as an exploratory tool only in our population for two reasons. First of all, LCA can be useful to identify subgroups, but there is no ‘true’ model or number of subgroups you can detect. The best fitting model can only be estimated based on the available measures and approximates the true subgroups that might be present. Second, LC membership assignment is a statistical procedure based on the posterior probability, with concordant errors of the estimation, rather than a clinical value that can be measured or evaluated. Therefore, we decided to use our LCA only in an exploratory tool, and perform our statistics using the actual phenotypes that predict LC membership and the associated genotypes.

Overall, this method worked well to differentiate the two subgroups present in our dataset. However, outliers were observed. A qualitative analysis of these outliers is available in the online supplementary data.The genetic substrate for the two phenotypic clusters can be discussed based on multiple experiments. Overall, we hypothesise two genetic clusters. One that is due to haploinsufficiency and one that is due to abnormal truncation of the activator.

The hypothesised cluster of variants that produce haploinsufficiency is mainly based on the experimental data that confirms NMD in two variants and the NMD prediction of other nonsense variants in Alamut. For the frameshift variants, it is also likely that the cleavage of the zinc finger domain results in functional haploinsufficiency either because of a lack of signalling domains or similarly due to NMD. Missense variants could cause haploinsufficiency through the suggested mechanism by Krauss et al who have illustrated that missense variants in the MID1 domain hamper the functional interaction with the MID1-α4-PP2A complex, leading to a subcellular location of GLI3.24 The observed missense variants in our study exceed the region to which Krauss et al have limited the MID-1 interaction domain. An alternative theory is suggested by Zhou et al who have shown that missense variants in the MBD can cause deficiency in the signalling of GLI3A, functionally implicating a relative overexpression of GLI3R.22 However, GLI3R overexpression would likely produce a posterior phenotype, as determined by Hill et al in their fixed homo and hemizygous GLI3R models.15 Therefore, our hypothesis is that all included missense variants have a similar pathogenesis which is more likely in concordance with the mechanism introduced by Krauss et al. To our knowledge, no splice site variants have been functionally described in literature.

However, it is noted that the 15 and last exon encompasses the entire activator domain, thus any splice site mutation is by definition located on the 5′ side of the activator. Based on the phenotype, we would suggest that these variants fail to produce a functional protein. We hypothesise that the truncating variants of the activator domain lead to overexpression of GLI3R in SHH rich areas. In normal development, the presence of SHH prevents the processing of full length GLI34 into GLI3R, thus producing the full length activator. In patients with a truncating variant of the activator domain of GLI3, thus these variants likely have the largest effect in SHH rich areas, such as the ZPA located at the posterior side of the hand/footplate.

Moreover, the lack of posterior anomalies in the GLI3∆699/- mouse model (hemizygous fixed repressor model) compared with the GLI3∆699/∆699 mouse model (homozygous fixed repressor model), suggesting a dosage effect of GLI3R to be responsible for posterior hand anomalies.15 These findings are supported by Lewandowski et al, who show that the majority of the target genes in GLI signalling are regulated by GLI3R rather than GLI3A.44 Together, these findings suggest a role for the location and type of variant in GLI3-mediated syndromes.Interestingly, the difference between Pallister-Hall syndrome and GLI3-mediated polydactyly syndromes has also been attributed to the GLI3R overexpression. However, the difference in phenotype observed in the cases with a truncating variant in the activator domain and Pallister-Hall syndrome suggest different functional consequences. When studying figure 1, it is noted that the included truncating variants on the 3′ side of the cleavage site seldomly affect the CBP binding region, which could provide an explanation for the observed differences. This binding region is included in the Pallister-Hall region as defined by Johnston et al and is necessary for the downstream signalling with GLI1.10 11 23 45 Interestingly, recent reports show that pathogenic variants in GLI1 can produce phenotypes concordant with Ellis von Krefeld syndrome, which includes overlapping features with Pallister-Hall syndrome.46 The four truncating variants observed in this study that do affect the CBP but did not result in a Pallister-Hall phenotype are conflicting with this theory. Krauss et al postulate an alternative hypothesis, they state that the MID1-α4-PP2A complex, which is essential for GLI3A signalling, could also be the reason for overlapping features of Opitz syndrome, caused by variants in MID1, and Pallister-Hall syndrome.

Further analysis is required to fully appreciate the functional differences between truncating mutations that cause Pallister-Hall syndrome and those that result in GLI3-mediated polydactyly syndromes.For the clinical evaluation of patients with GLI3-mediated polydactyly syndromes, intracranial anomalies are likely the most important to predict based on the variant. Unfortunately, the presence of corpus callosum agenesis was not routinely investigated or reported thus this feature could not be used as an indicator phenotype for LC membership. Interestingly when using only hand and foot phenotypes, we did notice a higher prevalence of corpus callosum agenesis in patients with posterior phenotypes. The suggested relation between truncating mutations in the activator domain causing these posterior phenotypes and corpus callosum agenesis was statistically confirmed (OR. 8.8, p<0.001).

Functionally this relation could be caused by the GLI3-MED12 interaction at the MBD. Pathogenic DNA variants in MED12 can cause Opitz-Kaveggia syndrome, a syndrome in which presentation includes corpus callosum agenesis, broad halluces and thumbs.47In conclusion, there are two distinct phenotypes within the GLI3-mediated polydactyly population. Patients with more posteriorly and more anteriorly oriented hand anomalies. Furthermore, this difference is related to the observed variant in GLI3. We hypothesise that variants that cause haploinsufficiency produce anterior anomalies of the hand, whereas variants with abnormal truncation of the activator domain have more posterior anomalies.

Furthermore, patients that have a variant that produces abnormal truncation of the activator domain, have a greater risk for corpus callosum agenesis. Thus, we advocate to differentiate preaxial or postaxial oriented GLI3 phenotypes to explain the pathophysiology as well as to get a risk assessment for corpus callosum agenesis.Data availability statementData are available upon reasonable request.Ethics statementsPatient consent for publicationNot required.Ethics approvalThe research protocol was approved by the local ethics board of the Erasmus MC University Medical Center (MEC 2015-679)..

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