Greys Anatomy, NCIS, The Voice, and other top shows have been promoting their season premieres for the past few weeks. ABCs hit reality show Shark Tank kicks off Season 11 this Sunday, September 29, with the infamous Mr. Wonderful putting a little bait in the water for viewers.

This season is already gearing up to showcase competitive waters, with several new and returning guest sharks. Among them are three women who have launched massively successful businesses and are now making their first venture in the tank.

Anne Wojcicki, CEO andco-founder of 23andMe,graduated from Yale University and became a Wall Street analyst. The business owner left finance to start medical school before she co-founded her now highly successful genetics company.

Katrina Lake, founder andCEO of Stitch Fix, created her company in 2011 with the goal of providing consumers with an innovative way to find their ideal clothes,according to ABC News. She succeeded, where now more than 3 million people utilize her styling service.

Known for her powerful athleticism on the court, world-renowned tennis champ Sharapova is also a savvy business owner. Creating thepremium candy line Sugarpova, the tennis star is an advocate of women in business, investing in several female-owned companies and mentoring women entrepreneurs.

In addition to the fierce females in the tank,founder and executive of KIND Daniel Lubetzkywill also be joining the sharks, making his first appearance in the season premiere.

Millionaire investor and fixture on Shark Tank Kevin OLeary, also facetiously called Mr. Wonderful, has landed some of the shows most successful deals. Known for his brash demeanor and love of all things money related, OLeary is extremely selective with which entrepreneurs he will partner.

So its all the more tantalizing for his followers to see his latest tweet promoting the shows premiere. Were baaack kiddies! @ABCSharkTank premieres THIS SUNDAY, OLeary posted on Twitter. I cant tell you much, but I make what could be my favorite deal in Shark History in the first episode! You dont want to miss this. #noseriously#setyourPVR.

Fans of the show realize that any deal that ranks as a favorite of Mr. Wonderful means the potential to make big profits, though the Shark Tank investor is always aware of risk. Ive had many, many successes on Shark Tank and many, many failures, he told CNBC last year. Thats the nature of venture investing.

Last month, the millionaire business owner made news due to a boating crash involving him and his wife Linda on Lake Joseph in Ontario, Canada. Two people were killed and three injured.

Late Saturday night I was a passenger in a boat that had a tragic collision with another craft that had no navigation lights on and then fled the scene of the accident, OLeary said in a statement on August 28,according to Us Weekly. I am fully cooperating with authorities. Out of respect for the families who have lost loved ones and to fully support the ongoing investigation, I feel it is inappropriate to make further comments at this time. My thoughts are with all the families affected.

Earlier this week, Linda OLeary was charged with Careless Operation of a Vessel following an investigation by the Ontario Provincial Police, according to Entertainment Tonight. She is scheduled to appear in the Provincial Offences Court in Parry Sound, Ontario,on October 29.

Now that the authorities have concluded their thorough investigation, I have no further comment other than to say that our thoughts and prayers are with the friends and families of those who lost loved ones in this awful tragedy, a spokesperson for OLeary told Entertainment Tonight. Our hearts go out to them.

The outcome of the October court date is sure to be detailed in upcoming news headlines.

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'Shark Tank's' Kevin O'Leary Just Teased the Show's Season Premiere With This Interesting Tweet - Showbiz Cheat Sheet

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Fordecades, it has been known that the mutations in the tumor suppressor genes BRCA1 and BRCA2 are associated with high risks of breast and ovarian cancer,but a new prospective cohort study provides the strongest evidence to date thatthese mutations are associated with the development of prostate cancer (PCa).

TheBRCA2 mutation appears to be morestrongly associated with PCa development than the BRCA1 mutation. In addition, the increased risk of PCa varies byfamily history of the malignancy and the location of the mutation within thegenes.

Ourstudy is unique in that we have recruited healthy men across the UK and Irelandwho have hereditary BRCA1 or BRCA2 mutations, and then followed themprospectively for up to 17 years to see if they would develop prostate cancer,said corresponding author Tommy Nyberg, a PhD candidate at the Centre forCancer Genetic Epidemiology at the University of Cambridge in the UK.

Thestudy, which was published in EuropeanUrology, included 376 male BRCA1 mutationcarriers and 447 male BRCA2 mutation carrierswho were identified in clinical genetics centers in the United Kingdom and Ireland.Of these, 16 BRCA1 and 26 BRCA2carriers were diagnosed with PCa during follow-up. Nyberg and his colleagues foundthat the risk of PCa is more strongly influenced by BRCA2 than BRCA1 mutations.The BRCA2 mutation is associated witha nearly 4.5-fold increased risk of PCa, whereas the BRCA1 mutation is associated with an approximately 2.4-foldincreased risk. This translates into estimated absolute lifetime risks fordeveloping prostate cancer of 60% for BRCA2and 29% for BRCA1 mutation carriers.We also found an association with more aggressive prostate cancer for men with BRCA2, but not BRCA1 mutations, Nyberg told Renal& Urology News.

Forthe men with BRCA1/2 mutations, therisk was greater for those from families where several family members had beendiagnosed with PCa than for those without such a family history. This probablyreflects the complex genetic landscape of prostate cancer susceptibility, withseveral genetic variants besides BRCA1/2mutations being known to influence the risk, Nyberg said.

Amongcarriers of the BRCA2 mutation, therisk of PCa increased nearly 1.7-fold with each relative diagnosed with PCa.Compared with the general population, BRCA2mutations in the so-called ovarian cancer cluster region (bounded by positionsc.2831 and c.6401) were associated with a nearly 2.5-fold higher incidence ofPCa, a lower risk increase than for mutations elsewhere in the BRCA2 gene. BRCA2 mutations outside this region were associated with a 5.9-foldrelative risk of PCa. Additionally, the BRCA2mutation was associated with a 5-fold increased incidence of Gleason score 7PCa and 3-fold increased incidence of Gleason 6 or less PCa. The mutation alsowas associated with almost 3.9-fold increased incidence of PCa mortality.

Isee the primary clinical application of our research as facilitating geneticcounseling and the early detection of prostate cancer, Nyberg said. Men whoare discovered to carry a hereditary BRCA2mutation, even if currently healthy, are at considerable risk of developingprostate cancer during their lifetime. A greater understanding of genetic riskvariants is continuously occurring, and consequently genetic counseling forprostate cancer is getting more and more accurate.

AnthonyV. DAmico, MD, PhD, Chief of the Division of Genitourinary Radiation Oncologyat Dana-Farber Cancer Institute and Professor of Radiation Oncology at HarvardMedical School in Boston said drugs already are available that target BRCA2 mutations. Studies are needed inmen who harbor BRCA mutations to investigate whether these drugs such as PARPinhibitors and platnium based chemotherapy can reduce the risk of metastasisand death from prostate cancer, Dr DAmico said.

Moreover,the new findings support recommendations that men with a significant familyhistory for PCa, especially those with multiple first-degree relatives with PCa,undergo genetic testing for the BRCA2mutation and then to be seen by a genetics counselor to be considered for screening at an earlier age than recommendedin standard guidelines.

Themajor implication here is that men with BRCA2in particular are at a significantly increased risk of developing clinicallymeaningful prostate cancer, and this risk might be influenced by factors suchas family history and the type of mutation that is inherited, said Amar U.Kishan, MD, Assistant Professor of Radiation Oncology at the David GeffenSchool of Medicine of UCLA.

Althoughthe therapeutic implications of the new findings are unclear, it istheoretically possible that men with mutations in DNA repair genes may derivebenefit from drugs such as poly (ADP-ribose) polymerase (PARP) inhibitors, butthe data to support such a strategy are limited to patients with advanced,metastatic castration-resistant PCa. In this setting, olaparib and rucaparibare approved for men with BRCA1/2mutant-tumors, though these mutations can be either inherited or restricted tothe tumor, Dr Kishan said. Whether men with an inherited BRCA2 mutation, whodevelop an aggressive but early stage prostate cancer, would benefit from thistype of therapy, in combination with surgery or radiotherapy, is not known.Several studies are investigating this concept.

ToddMorgan, MD, Associate Professor of Urology and Chief of the Division ofUrologic Oncology at the University of Michigan in Ann Arbor, said the new studyadds important data to help guide patient counseling and may allow for improvedearly detection strategies in men with BRCA1/2 mutations. At his institution, DrMorgan and his colleagues have implemented an early detection clinic for menwith BRCA1 or BRCA2 mutations, which is modeled after similar clinics for femalecarriers of these mutations (https://www.rogelcancercenter.org/cancer-genetics/prostate-cancer-risk-clinic).

Medicaloncologist David Wise, MD, PhD of NYU Langone Health in New York, said the newfindings may change the conversation for men carrying the BRCA2 germline mutation. Based on this study and others, newguidelines are needed to personalize prostate cancer screening for men carryingthe BRCA2 germline mutation. Clinicaltrials testing PARP inhibitors, already FDA approved for ovarian and breastcancer, are ongoing in BRCA2-associatedprostate cancer. Based on promising data from these clinical trials, the FDAhas granted breakthrough therapy status for two PARP inhibitors, rucaparib andolaparib, for men with castration-resistant prostate cancer, Dr Wise said.

Reference

NybergT, Frost D, Barrowdale D, et al. Prostate cancer risks for male BRCA1 andBRCA2 mutation carriers: A prospective cohort study [published online September5, 2019]. Eur Urol. https://doi.org/10.1016/j.eururo.2019.08.025

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Prospective Study Characterizes PCa Risk Linked to BRCA1, BRCA2 Mutations - Renal and Urology News

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Three wolves found a new home at Isle Royale National Park a remote island cluster on Lake Superior by the Upper Peninsula bringing the islands wolf population to 17.

The goal of the Isle Royale wolf translocation effort is to have at least 20 new wolves moved to the island over a three to five year period. The park is in year two.

Last year, park officials translocated wolves from Minnesota and Ontario to repopulate the island.

Isle Royale National Park Public Information Officer Liz Valencia says this year seven wolves were caught in the Upper Peninsula for translocation, but only four were deemed fit for life on the island. Researchers conduct field tests while catching the wolves to determine if they are a good match, she says.

The four, two males and two females, were brought to the park during the week of Sept. 9, but one female died within the first few days on Isle Royale.

Valencia says the researchers wanted to bring in wolves from Michigan to widen the genetic pool on the island, which would help with the long-term sustainability of the population.

Once the wolves are on the island there is limited opportunity for wolves to come and go. And also limited opportunity to bring new genetics in, so they recommended bringing in wolves from a variety of different places, she says.

Researchers plan to study the wolves over the next few months with a focus on the social organization and reproduction of the wolves, Valenciasays.

The rest is here:
3 more wolves added to Isle Royale on Lake Superior - Interlochen

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Seedstock producers might have one of the toughest jobs in the beef industry as theyre tasked with raising quality animals that can go down several different avenues. Whether it be into commercial herds or registered, the resulting animals need to be able to perform.

Several influencers sat down on a panel at the first Cattle U event in Dodge City, Kansas, at the United Wireless Arena July 31. Cattle U was sponsored by High Plains Journal. The seedstock panel included Glen Klippenstein of Missouri; Lee Leachman, Leachman Cattle of Colorado; Rick Pfortmiller, Neogen; Kelli Retallick, director of genetic services at Angus Genetics Inc.; and Tom Strahm, commercial marketing director for the American Gelbvieh Association. The panel was moderated by Lorna Marshall of Select Sires.

What are the top factors that you think influence bull purchases the most, today and in the next five years?

Journal photo byKylene Scott.

Strahm said a couple of traits come to mind for him.

Well, I think two of them would be calving ease and growth right now, he said.

Pfortmiller, who works for a genetics company, said choosing the most important traits depends on what the beef producer is going to do with their resulting calves.

As we interact with customers, we hear a couple of things, he said. One would depend on where theyre kind of coming from, in terms of their cowherd, what their focus is.

If the producer is strictly in the business to sell calves right off the cow, traits like longevity and docility rise to the top.

Docility is one that we get thats justified as more of our customers are getting a little longer in the tooth, Pfortmiller said. Then the other piece that I think is just all that is an increase of predictability. People are looking at any of the information thats coming in, and how can I reduce my risk?

Leachman said producers are paying attention to indexes that help predict profitability in the cattle.

Thats probably the No. 1 driver, maybe even more than any other component traits, Leachman said. They probably have gone to worrying less about how they get there, and more about how much profit they generate.

And that surprises Leachman, because if you look at the industry as a whole, yearling weight tends to be a big driver.

But in our model, its not a big driver, he said. Our customers are pretty much not worried about buying lower growth cattle, which I found surprising, and that we put more emphasis on explaining the difference between the maternal index and just went profitability from birth to weaning versus weaning to yearling, weaning to harvest.

What technology is going to be most applicable for commercial cow-calf producers in the next five years?

Pfortmiller is both a commercial producer and works in genomics and believes the holy grail is efficiencyand just not feedlot efficiency.

The notion of cow efficiency out there on the range, the things that I think were going to learn about animal behavior, and adaptability and all of that, Pfortmiller said.

However, theres not many good phenotypic databases out there with the genomic companies.

We get approached nearly every day by groups, small breeds, other groups wanting us to find a DNA test, he said.

Journal photo byKylene Scott.

But that requires a couple thousand phenotypic records of cows who are all the same age, raised the same way and raised in the same environment. Thats nearly impossible.

Strahm agrees. He knows of a commercial producer whos selected traits that are important to him and his production system and environment. He selected certain traits and had done a considerable amount of DNA testing, which he used to select replacements. His home-raised replacements go back into the herd and the others are fed out. The producer has a very extensive data set for his Balancer bulls and cows.

Recently, he took 10 years worth of data from feeding his own cattle, put it all on one grid to keep the value and price constant, Strahm said. And he increased the value of those cattle $300 per head for a 10-year period, and primarily focused on improving marbling, improving growth, and maximizing carcass growth.

Strahm was impressed he was able to market those cattle and get top dollar.

Then to be able to market those cattle in the end and keep everything in check, and take a balanced approach so that you can keep those replacement females in his herd, he said.

From Marshalls perspective at Select Sires, she thinks producers need to use the tools that are already available.

I think sometimes we leave a lot of money on the table, we dont pay attention to dollar values or selection indexes, or we dont pay attention to the EPDs, maybe as much as we should have, Marshall said. Im taking advantage of things even as simple as parentage testing.

In pastures with multiple potential sires and trying to avoid inbreeding, it gets hard to determine which bulls arent doing their job. Using a genetic test to determine exactly which animals are from each bull can be really valuable to the commercial producer.

When we think about things like Gene Max Advantage, and some of the other commercial heifer replacement tests that are out there, theres things out there right now that we can capitalize on, Marshall said. I know when we talked about the cost per test, $28, it seems like enough of an upfront cost test for your replacement females to find the best ones. But when we think about how much it costs to actually build that replacement female, right to get her into production, you know, $28 actually seems like a pretty incremental piece of cash.

Leachman sees the benefit of DNA tests to help predict which cows are going to stay in the herd for a long period of time.

If our DNA tests can predict that, thats something you should go home and use, because right now, you pick those heifers and there are differences in productivity, Leachman said.

There are tests that can help predict calf performance and weaning weights on the cows progeny, but its still hard to predict a cows longevity.

What we cant predict really well is whether theyre going to stay there and be there, Leachman said. I think we all know that when we go out in our herd, and we look at these 10-year-old cows that have had nine calves, that theyre making us money. We also know that when we go out after preg check, and we get rid of those young females that are open, they didnt make us money.

Leachman believes a tool is close that will help producers find out which heifers have a higher propensity to end up in the 10-year-old category instead of the open two and three and four.

I think that we need to be testing that tool and putting it to work, Leachman said.

What would be the most important thing a commercial cow-calf producer needs to look for in their seedstock provider?

Klippenstein shared a quote, in life, business, and politics its one third motion two-thirds promotion. And in order to succeed in the beef industry, one has to promote.

Otherwise, youd never get a chance, he said. Which is the most important thing to develop a positive reputation.

Klippenstein said he always aimed to raise good bulls that would make his customers money.

The sum total of all kinds of things that all of us, any one of us, who are in the cow business can do, if we want to, Klippenstein said.

In his time in the business, hes seen some magnificent herds from guys who just went through high school, but knew exactly what kind of cattle they wanted and spent time with the herds.

Now weve got all these tools, he said. Lets go to the people who have the reputation and understand the use of the tools and say this is what I want to do with my genetics with bulls I buy from you.

Pfortmiller said it comes down to one thingintegrity.

Find a seedstock supplier that is managing their cattle in a way that youre trying to manage them, he said. Theres all kinds of tools that you can use, but if you start with those two things youre going to be pointed in a pretty good direction.

In the future, Pfortmiller believes transparency will stay at the top of mind.

I think the technologies we hear aboutthe block chain and all of thatI think were going to have a greater opportunity to transfer that transparency to all the related parties.

Leachman echoed those sentiments, and believes the biggest thing cattle producers are going to have to deal with in the next three years, is something thats been lurking in the shadows for a while.

Maybe we dont think about it enough, but seems like the last time the dairy herd had to liquidate we had a significant disruption in the market, Leachman said.

With the alleged animal abuse videos coming from large dairies, Leachman thinks it will influence how consumers see beef producers even though the industries are different.

Were not dairies, but those videos are what people think about us, he said. Theyre coloring how they think we treat our animals. Its a big problem for us, because we are going to get judged by the rotten apples that are lying in the grass. And I think its a big risk right now.

He believes were dangerously close to seeing a massive reshuffle in the dairy industry, that could hurt the beef industry in the short run.

It could change the way calves are being raised on dairies and theyre precarious enough at this point that this would push a bunch of dairies out of business, Leachman said.

For Retallick, picking a seedstock supplier that aligns with breeding goals and objectives is vital.

If you are selling your calves and weaning and finding those seedstock producers that are really developing their bulls and aiming their genetic potential towards that specific environment versus are you retaining ownership on those animals through the feedlot and hanging them on the rail, she said. And really trying to approach your seedstock provider as a person who really aligns with your own personal goals.

Also create your own breeding objective first, then knowing where you want to go and create that plan.

I think one of the biggest things we have to do is stick with the plan, Retallick said. I see it in the seedstock industry more probably than in the commercial industry, but in the seedstock industry it seems like we can really chase things. We can chase the biggest number.

Cattle producers should stay disciplined and believes some of the most successful seedstock producers are the ones who have stuck to their goals and objectives for their operations.

It doesnt matter what your breeding goal or breeding objective is as long as its making you profit and its making your customers profit, she said. But make sure you have a breeding goal, and stay disciplined to it. Stick with it.

She said when incorporating genetic or tech tools, remain disciplined and carry through with themeven when the data doesnt show what a producer was wanting or expecting.

Then they dont reap the full benefits of something like that and so if you can remain consistent I think and disciplined, I think thats one of the biggest things we can kind of do in all segments of the industry, Retallick said.

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Influencers eye the future of seedstock industry | Livestock - High Plains Journal

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TONING is not real.

It is believed to be a process of exercise, which will result in a toned body a fit-looking body that is not excessively muscular.

This process is supposedly brought about by exercising the muscles, including those in trouble areas, with the expressed goal of not making them bigger but tighter and firmer.

This concept is a physiological impossibility. The idea that muscle will become tight and less jiggly is not how muscles develop.

Muscles will not become firmer in your day-to-day life because of a few exercises. The development of muscles have a few variances, which are perceivable:

Bigger or smaller through hypertrophy (growth) or hypotrophy (shrinkage);

Stronger or weaker;

More conditioned (accommodate for shorter recovery time, greater endurance ability and speed) or less conditioned;

More effective stretching or shortening.

Toning was born out of the fear expressed by women, that although they didn't want to have unwanted jiggling, they did not want to become muscular and manly-looking, or lift heavy weights either.

Gym owners, exercise instructors, trainers, and coaches assured these women that they could tighten their muscles without making them bulky, by using lighter weights, body weight, bands with light to medium resistance, higher repetitions, and at times incorporating more movement, perhaps aerobic or dance.

The facts

Flexed muscle is firm. Relaxed muscle, being mostly water, is jiggly. Fat is jiggly, tight skin is tight, and stretched out or weakly elastic skin is loose. Finally, bone is rigid.

So ultimately it comes down to what you are made of between your bones and skin and the elasticity of your skin.

If you have little body fat:

And have never stretched out your skin, your body will appear tight (non-jiggly, toned), even if you have very little muscle;

And large muscles and your skin is tight, you will appear tight;

But have stretched skin and little muscle, you will appear jiggly and loose.

If you have excess body fat and have damaged the connection between your skin and your muscle's fascia (the sheet of connective tissue which acts like a glue between skin and muscle) then:

If your skin is still elastic and tight, you may not appear jiggly;

If your muscles are smaller, you may appear more jiggly;

If your muscles are larger, you will appear less loose and jiggly.

If you are losing fat, lessening the fat under the skin:

Less muscle will naturally mean less utilised volume between the skin and more, resulting in a loose jiggly appearance;

Increased muscle will be needed to occupy as much of that space left from the loss of fat to avoid a loose jiggly appearance and feel.

Take-away message

a. Little muscle, little body fat but stretched skin, you will appear jiggly and loose.

b. Excess fat, less muscle, you will appear more jiggly.

c. Larger muscles, less jiggly (this is why tummy tucks are often the only solution for the waist. Often, a lot of fat is lost, but the muscles of the waist do not grow and occupy a much greater relative volume.

When losing fat, increased muscle will be needed to occupy as much of that space left to avoid a loose jiggly appearance and feel.

More facts

The only solutions to avoid jigglyness are:

1. Tighter skin, which only youth, genetics and surgery can significantly provide. Do not waste time and money believing that products will help.

2. More subcutaneous fat: Worst solution ever fated to be a long-term disaster.

3. More muscle: A solution filled with a world of benefits.

There is only one effective way to significantly increase muscle and this is through progressive resistance training. You have to activate the muscle under tension, resulting in (perfectly natural) microscopic tears, which will heal with nutrition and rest, resulting in muscle hypertrophy (growth).

It is proven that this is most effectively attained with resistance, which the individual will consider heavy. So heavy, in fact, that instead of dancing or hopping around with the weight, they may only be able to accomplish three to 10, perhaps 12 repetitions, essentially training like a body-builder, as, of course, that is what is needed for muscle building.

Do not fear, if your goal is to avoid a manly, bulky appearance, rest easy. That appearance takes more time and dedication than you can likely find, and often still, only with the use of anabolic steroids.

If you are a female, you can rest even easier, that bulky look requires a level of testosterone that the very great majority of women can never have.

So, as hard as you try, at best you will simply look the way one should athletic, healthy and fit.

Fitz-George Rattray is the director of Intekai Academy, which is focused on helping people live a healthy lifestyle through nutrition and weight management. If you are interested in losing weight or living a healthier lifestyle, give them a call at 876-863-5923, or visit their website at intekaiacademy.org.

Now you can read the Jamaica Observer ePaper anytime, anywhere. The Jamaica Observer ePaper is available to you at home or at work, and is the same edition as the printed copy available at http://bit.ly/epaperlive

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That toning myth - Jamaica Observer

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The idea of paying a company hundreds of thousands of dollars to freeze your body or brain in the hope that they will one day bring you back to life initially may seem farcical.

The mere fact that this notion has featured in several movies and television shows suggests this is make-believe, something that would probably not happen in real life.

Who could forget that fantastic scene from Austin Powers when doctors successfully thaw out Mike Myers so that he can live in another era?

Well, this is no movie. And no joke.

And interestingly, Alcor CEO Max More isn't bothered by cryonics being trivialised for entertainment.

"I don't think it stigmatises it, I think it actually makes it seem more plausible," More says when I asked him about this at his Arizona Headquarters.

He shares the building with about 170 frozen bodies and brains all suspended upside down in liquid nitrogen-filled vats.

As I walked through the room where all these cryopreserved people are, it feels eerie and weird, quite frankly.

But there are more than 1250 Alcor members who've signed up for the same fate from all over the world Australia, Canada, Germany, Japan, Mexico, Portugal, the UK, the US, Italy, Israel and more.

These members all have the choice of freezing their whole body for about A$300,000 or just their brain for A$118,000.

That is, needless to say, a lot of cash to hand over for a service the company can't even promise will eventuate.

"There's no guarantee but it seems like it's worth a shot," More says.

Alcor founder Linda Chamberlain explains that members sign a nine-page consent form.

"We tell people everything that could possibly go wrong and that's extremely important because what we are doing is still experimental and people need to know that," she says.

"The Alcor Life Extension Foundation is the world leader in cryonics, cryonics research, and cryonics technology," their website reads.

"Cryonics is the practice of using ultra-cold temperature to preserve a human body with the intent of restoring good health when the technology becomes available to do so."

The hope or as they prefer to call it, "probability" is that one day science will have been developed that is capable of reviving frozen patients.

They would come back to life as the same person, with the same DNA.

I was wildly sceptical about this upon arriving at their Scottsdale building in the hot desert of Arizona.

I was worried members may be vulnerable to handing over thousands of dollars on false promises.

But it was quickly evident that those running this business whole-heartedly believe the plausibility of cryonics recreating life in the future.

The reality is not many people can afford to become a member and embark on such a thing.

But, if you were a millionaire, even if you thought this was questionable, what have you got to lose? When you're dead, you're dead.

Or are you? That's the question.

Even the masterminds behind this concept don't have the answer. Hence the consent form.

Continued here:
Inside the lab where frozen bodies wait for a second lease on life - 9News

Recommendation and review posted by Bethany Smith

The House Science, Space and Technology Committee said it is drafting its own nuclear energy bill, which two people familiar with the matter said will imperil passage of the Nuclear Energy Leadership Act in its current form. But people in talks with the committee say that it is likely to draw heavily from the premier advanced nuclear energy legislation in Congress.

House Science Committee majority staff said in an email Thursday that the committee is still in the process of drafting its legislation. Our bill wont be identical to NELA, but we expect it will comprehensively authorize and guide DOE nuclear energy R&D activities, the committee staff said.

Jeremy Harrell, managing director of policy at the conservative energy nonprofit ClearPath, said the science committees move does not necessarily extend the timeline or complicate passage of much of NELA, which would call on the federal government to establish a 10-year national strategy for nuclear energy and to demonstrate multiple advanced nuclear reactor concepts over the next several years, among other things.

He said the committee appears supportive of NELAs reactor demonstrations provision, for example, through which the Energy Department would need to demonstrate between four and seven advanced nuclear reactor designs by the end of 2035.

I think when push comes to shove, Science and Tech wants to make sure that they have their own bill within their circuit, Harrell said. Theyve got their own ideas; theyre not going to just take something from Lisa Murkowski, the Alaska lawmaker who chairs the Senate Energy and Natural Resources Committee and sponsored the bill in the Senate.

Science moving forward with its own bill means that another vehicle will need to be found to enact an industry-favored provision of NELA to extend the federal power purchase agreement length from 10 to 40 years for nuclear, since the provision falls outside the committees jurisdiction.

I cannot envision any legislation on power purchase agreements coming out of the House any time soon, said a person familiar with the matter, who said there has been no sign from Energy and Commerce that they intend to work on power purchase agreement legislation.

The person, who said the plan appears for the committee to circulate a working draft of the bill to stakeholders before Thanksgiving, said that the science panel is considering including some language on high-assay low-enriched uranium fuel which multiple advanced reactors would require to operate from NELA as well as from the McNerney-Flores HALEU bill that recently passed the House.

The House Science measure, which is intended to lie under the panels sole jurisdiction, means that movement on NELA in the House is now more unlikely, according to two people familiar with the developments.

Alyse Huffman, the committees American Nuclear Society Glenn T. Seaborg congressional fellow, is the lead author of the new legislation. It remains unclear whether the panels nuclear bill will be folded into a broader energy package out of the committee.

In a brief interview Friday morning, House Science EnergySubcommittee Chairman Conor Lamb (D-Pa.) said the committee is considering a possible parallel to the Senate version of NELA, while somewhat separately considering legislation for advanced nuclear research in areas such as the national labs and ARPA-E program. He did not yet have a sense of when the committees legislation might come out and said that talks were ongoing with House Science Chairwoman Eddie Bernice Johnson (D-Texas).

Were looking at things for life extension of the existing plants, which is important, Lamb added, as NELA instead focuses on newer reactor concepts.

It is not yet clear whether the House Energy and Commerce Committee will take separate action on NELA. The bill has also been referred to the House Armed Services and Oversight and Reform committees.

The only thing Im aware of is the positive news that Representative Clyburn is very interested in advancing it, Rep. Elaine Luria (D-Va.) said in a brief interview Friday morning, referring to the Democratic representative from South Carolina. Luria introduced NELA in the House on June 18. Were working with his staff to help that happen.

Lamb said Luria had asked him to champion NELA within the science committee.

An Energy and Commerce Committee spokesperson said in an email Friday that there have been staff-level conversations between it, Luria and others.

David Blee, chief executive of the U.S. Nuclear Industry Council, cautioned that NELA has not yet passed the Senate a difficult feat in the current political climate.

Harrell said that the current expectation is that senators will try to pass NELA as part of a larger energy package, with potential floor time in January or February.

Blee said that the preference is usually to have it as part of a package, since both parties want action across the energy and environmental space, and the House will likely want balancing legislation for renewables and other energy sources if it moves on a nuclear bill.

To the extent the House Science panel comes up with that recipe, thats helpful, Blee added.

Even if NELAs provisions are not enactedquickly, there are promising signs from the Senate that bolstering federal support and development of advanced nuclear technologies is a priority. The Senate Energy and Water appropriations subcommittee, led by NELA co-sponsor Lamar Alexander (R-Tenn.), has slated key funding for provisions within NELA without cutting other federal nuclear energy R&D efforts, as had been feared. Harrell said the House is also likely to be supportive of nuclear energy funding.

But a lot remains up in the air on the budget and appropriationsas a whole, and it remains useful to pass legislation authorizing the provisions of NELA outright, said Harrell, especially since the demonstration portion runs through 2035.

We think its critical that were demonstrating new technologies in that time frame, Harrell said. Otherwise, nuclears going to get left behind.

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Path Shifts for Advanced Nuclear Legislation in the House - Morning Consult

Recommendation and review posted by Bethany Smith

Growing emphasis on the food safety and longer shelf life has played an important role in the development of ingredients that aid in food preservation. These ingredients vary from simple water content to salt or sugar to chemicals like antioxidants and are used to prevent growth of microorganisms, thereby delaying the spoilage process. In terms of origin, food safety and shelf life extension ingredients can be synthetic or natural in nature.

Food preserving ingredients have been an integral part of kitchen aisles in the form of lemon, ginger, vinegar, spices, salt and sugar. Their traditional utilization was replaced by synthetic ingredients with increasing commercialization of the food industry in past decades. However, with the dissemination of knowledge related to harmful effects of synthetic ingredients, currently, the industry is witnessing a prominent shift toward natural ingredients for food safety and shelf life extension.

Shelf Life Extension Ingredients Market Notable Developments In January 2019, Kemin, a global nutritional ingredient company launched NaturCEASE Dry, a new clean-label and natural food safety solution. The product is a combination of natural plant extract and buffered vinegar for the use in the preservation of processed meat products. Researchers from the Penn State University studied a class of Alkylresorcinils (AR) found in grain plants such as barley, rye and wheat. Scientists revealed that these compounds can act as food preservative owing to their antioxidant and mold and bacteria prevention properties. The European Court of Auditors (ECA) announced publication of the report 'Chemical hazards in our food: EU food safety policy protects us but face challenges on 15 January 2019. The conclusion of the report focuses on updating current EU legal framework and implementing and enforcing new issues that compromise consumer protection from chemical hazard in food.

Shelf Life Extension Ingredients Market Dynamics

Clean-Label Trend Fuels Synthetic to Natural Transition in Food Ingredient Landscape

Browse more detail information about this report at https://www.tmrresearch.com/shelf-life-extension-ingredients-market

Naturally sourced ingredients have gained significant traction as consumer preference for natural products continues to surge. In terms of effectiveness, natural preservatives are superior in delivering greater protection and longer shelf life. As they work with equivalent efficiency and are healthful in nature, adoption of naturally sourced ingredients is increasing consistently as compared to the synthetic options.

Natural ingredients such as antimicrobials or antioxidants have additional potential health benefits also. Well aware of the increasing consumer demand for natural food products that are without artificial ingredients, manufacturers in the food ingredient market are introducing bio-based or naturally sourced food safety ingredients.

Frozen Foods Drive Demand for Specialized Food Safety Ingredients

Ranging from salads to sauces or ready meals to rice, a plethora of food products are available in frozen forms. As the demand for fresh and frozen foods increase across the globe, food manufacturers are seeking innovative ways to introduce novel food safety ingredients to extend the shelf life of frozen foods.

Manufacturers in the food safety and shelf life extension ingredient market are introducing ingredients specific to refrigerated products. Along with providing safety, these ingredients are label friendly and help in reducing sodium content while enhancing consumers sensory experience.

Shelf Life Extension Ingredients Market Regional Outlook

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North America presents lucrative opportunities for the Shelf Life Extension Ingredients Market on the back of buoyancy in regions the food and beverage industry and presence of leading F&B companies.

The market is likely to witness increasing opportunities in the developing countries of Asia pacific. These countries are witnessing huge demand for frozen foods, RTD food and beverages and processed food, thereby presenting higher potential for the market in the future.

Shelf Life Extension Ingredients Market Segmentation

The Shelf Life Extension Ingredients Market is segmented into following,

Based on type, Shelf Life Extension Ingredients Market can be segmented in, Natural Synthetic

Based in function, Shelf Life Extension Ingredients Market can be segmented in, Anti-oxidant Anti-microbial Others

Based on application, Shelf Life Extension Ingredients Market can be segmented in, Dairy Products Snacks Meat & Poultry Products Beverages Bakery Products Others

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Shelf Life Extension Ingredients Market To Grow in the Coming Years, New Research explores Factors Responsible - NewsStoner

Recommendation and review posted by Bethany Smith

Fall is one of the biggest seasons for publishers, so there are always lots of new books to get excited about. But how many of these books exist outside of the white, cisgender, heterosexual norm? The answer continues to be, not enough.

However, there are still a lot of compelling titles being released over the next few months from LGBTQIA+ writers for those of us who seek broader representation in literature. Theres a sports anthology that bursts with poetry and a decolonization narrative; there are memoirs about aging; short stories; novels that take place in Uruguay and Portland, Oregon,and Brooklyn, New York; new books from debut authors and old favorites of the queer literary canon. Its an exciting time for queer readers: Although we have a long way to go toward diversifying the publishing industry, more of our stories are being given space to be told. Here are some highlights coming up.

Elissa Altman, Motherland(Ballantine, 8/6)

Elissa Altman is the author of two previous memoirs, including Poor Mans Feast: A Love Story of Comfort, Desire, and the Art of Simple Cooking, born of her James Beard Award-winning blog of the same name. In this new memoir, Altman explores her codependent relationship with her mother, a glamorous Manhattan singer named Rita. Growing up, Elissa didnt fit her mothers high-femme standards of makeup, dresses, and generally keeping up appearances. Her identity as a lesbian was also a source of tension between them. As an adult, Elissa moved to New England, where she lived with her wife, Susan, forging her own identity away from her mother. But when Rita hasa fall that results in physical disability, Elissa finds herself taking care of her mother again and coming to terms with the reality of caring for an aging parent. With poignant and often very funny prose, Motherland makes for a wonderful addition to the narrow canon of stories about queer women caring for their aging mothers. Another great memoir in this vein is Cherre Moragas Native Country of the Heart (FSG, 2019).

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Kimberly King Parsons, Black Light (Vintage, 8/13)

If you pay attention to best-of book lists, Black Light has probably come across your radar.The prose in this debut short-story collection is richly detailed in describing both the characterswho are weird, flawed, and so humanand the settings theyre in; its lyricism feels like a delicate dance and a gut-punch at once. The book isnt marketed as queer, but there is a lot of queer content here; Parsons is particularly strong in writing about the complicated intimacies between teenage girls.

Carolina de Robertis, Cantoras (Knopf, 9/3)

The new novel from the author of The Gods of Tango (which also features a protagonist who explores their fluid gender and sexuality), Cantoras begins in 1970s Uruguay as five young queer women come together on an island for a getaway. Over the course of about 40 years, it follows their lives together and apart, including under the Uruguayan dictatorship during the 70s and early 80s. This is another multi character-driven epic, both painful and beautiful to read, with vivid prose. Also, it has one of the best opening paragraphs this reviewer has seen in a while.

Carley Moore, The Not Wives (Feminist Press, 9/10)

In this debut novel, Carley Moore (author of the essay collection 16 Pills) transports the reader right into the heart of the Occupy Wall Street movement. Brimming with details that will bring anyone who remembers the early 2010s right back to that era (from fashion, to the way people talked about politics and queerness), this story follows a sprawling cast of characters, almost all of whom are queer: the protagonist, Stevie, her young daughter Sasha, her friends Mel and Jenny, a young runaway named Johanna, and a number of others. Moore deftly captures the atmosphere of Zuccotti Park as well as the political atmosphere around the movement, which spurred people all over the world to renew discussions about class, race, gender, and social structures.

Gabby Rivera, Juliet Takes a Breath (Penguin Teen re-release, 9/19)

This debut novel from Gabby Riverawho also wrote Marvels Americawas originally published in 2016. An instant queer YA classic, it is now getting the wide distribution it deserves with a reprint from Penguin Teen. It follows maybe one of the best characters ever written, Juliet Milagros Palante, a young queer Latinx woman from the Bronx who spends a summer interning for fictional white feminist icon author Harlowe Brisbane in Portland, Oregon. The ways in which Juliet grapples with coming out, unlearning colonialist and white supremacist ideas, and discovering her own values give this novel teeth. But the real joy of this story is Juliet herself, a character whose self-doubt is rivaled by her self-confidence; whose big heart and book-nerd brain envelop the reader; and whose relationships with the fascinating characters around her make for one of this falls most satisfying reads.

Jacqueline Woodson, Red at the Bone (Riverhead, 9/17)

Jaqueline Woodson is an award-winning author of many YA novels, including Another Brooklyn and Brown Girl Dreaming. Shes now bringing her signature spare and poetic prose to this adult novel set in Brooklyn. The book opens with the seemingly lighthearted scene of 16-year-old Melodys coming-of-age ceremony, which happens to the orchestral tunes of Prince. From there, the story moves back and forth in time, spanning the events faced by three generations of Melodys family. Red at the Bone has one explicitly queer character, but isnt really about being queer. In that way, its a beautiful example of queerness being a part of everyday life, in families and friendships. The book touches on class, gender, sexuality, age, race in the United Statesall within a tender story of love in its many forms. Its a perfect one- or two-sitting read for a crisp fall day, one that will stay with you long after its over.

Jeanette Winterson, Frankissstein (Grove Atlantic, 10/1)

Jeanette Winterson is well-known to many queer readers from her prolific work, notably Oranges Are Not the Only Fruit, Written on the Body, The Passion, and Why Be Happy When You Can Be Normal? Her latest novel delivers a reinterpretation of Mary Shelleys Frankenstein. Multiple Frankensteins are being produced and experimented with in this novel:Shelley is creating her novel in the fictionalized chapters about her; scientist Victor Stein is creating an AI in the very near future, while trans nonbinary scientist Ry is falling in love with him; a creepy foundation in Arizona called Alcor Life Extension is scheming to create live humans from body parts; and a man named Ron Lord is creating the next generation of sex dolls. Its a wacky ride about what it means to be human and what it means to be alive, and it will not disappoint Winterson fans.

Natalie Daz & Hannah Ensor, eds., Bodies Built for Game (University of Nebraska Press, 10/1)

You might recognize Natalie Dazname from her professional basketball career, or her poetry collection When My Brother Was an Aztec (shes also got one forthcoming in March called Postcolonial Love Poem); and Hannah Ensors from her collection Love Dream With Television or her recent Lambda Literary 2019 Judith A. Markowitz Award for Emerging Writers win. But if you dont, rest assured that they are a dream team for bringing together this unique and innovative anthology of sports writing. With an array of writers (including Danez Smith, Fatimah Asghar, and Hanif Abdurraqib, among others), this book picks apart the meaning of sport from the perspective of people who are not cis straight white men. From participation in sports (team or otherwise) to sports fandom; from analyzing the metaphor of sport to examining modern sports oppressive, exploitative, and colonialist roots; from exploring the intersections of identity (including gender and sexuality) and sports to grappling with the corporeal demands of athletics; his anthology breaks all kinds of barriers. And although it includes essay, fiction, and creative nonfiction, the book centers poetry as the dominant form, which is unusual for sports anthologies. This is definitely a must-read before that NFL game on Thanksgiving Day.

Edie Windsor and Joshua Lyon, A Wild and Precious Life (St. Martins, 10/8)

Edie Windsors case against the United States led to the U.S. Supreme Court ruling that overturned the Defense of Marriage Act. This, in turn,paved the way for Obergefell v. Hodges, which legalized marriage equality nationwide in 2015. But Windsor was more than these headlines, and more than her constant presence at rallies and protests. This memoir, begun by Windsor and completed by her co-writer Joshua Lyon after her death in 2017, chronicles her life beginning with her childhood in Philadelphia and moving through her young adulthood in 1950s Greenwich Village, her life with her partner of 44 years, Thea Spyer, and her fascinating rise in the ranks of computing at IBM. Windsor was a queer woman who believed in her right to take up space and be seen, which makes for an uplifting and inspiring story in which Lyons does an excellent job of making sure Windsors stirring and joyful voice shines through.

Jaquira Daz, Ordinary Girls (Algonquin, 10/29)

Every once in a while a truly electric debut memoir comes along, and this fall, Ordinary Girls is it. As a young person, Dazlife was upended many times by physical and emotional violence. Longing for family and sometimes finding itin her sister, her grandmother, her friendsDaz writes her story in a musical prose that plays with future and past tense, putting the reader in the narrators shoes as she navigates unreliable memories and unknown futures. Daz narrative starts in Puerto Rico, goes to Miami Beach, into schools and juvenile detention centers, into stronger-than-blood relationships with friends, lovers and family; it goes into the navy, into familial legacy and colonial history. Its the story of an ordinary girl; its the story of all of the extraordinary girls. Daz is a skilled writer who strongly layers micro details with the macro structures of identity, white supremacy, colonialism, and brown, queer, and femme resilience and resistance.

Cyrus Grace Dunham, A Year Without a Name (Little, Brown, 10/15)

A Year Without a Name, an honest account of one persons journey with gender, identity, and mental illness, is Cyrus Grace Dunhams snapshot of a recent two-year span in which they become a changing adult in a changing world. Its a quick read, but punchynearly every sentence is sharp, full of importance, at once deeply intellectual and ethereal. Dunham navigates how confusing gender is: how useless it can be while also existing as an essential facet of identity. Theyre extremely self-aware, which at times feels like more of a burden than a gift. Dunham stays true to their unfinished story by packing a lot of meaning into just 176 pages but never reaching concrete conclusions. But the concrete would be antithetical to the story; Dunham lives in the truth that all of us are unfinished, forever growing and learning. This in itself is a very queer frame of thought.

Carmen Maria Machado, In the Dream House (Graywolf, 11/5)

Carmen Maria Machado became everyones gay aunt, thank goodness, when she Mary Poppinsd into our hearts with her genre-bending, brutal, gorgeous, queer, wild debut story collection, Her Body and Other Parties, in 2017. Her debut memoir, In The Dream House, is a firecracker of a follow-upa story that is as scary as it is playful, as intellectual as it is emotional, as personal as it is universal. Its the story of Machados relationship with an abusive partner and how it, along with her upbringing and beliefs imposed by society, affected her growth as a person. It brings to the page something that is sorely lacking in mainstream literature: the reality of abuse in queer relationships, especially lesbian ones. Machado uses her characteristic wit and fearlessness to shed light on love, terror, history, culture, narrative structure, and representation from multiple angles. Its a must-read, haunting story for the dark months ahead.

Tommy Pico, Feed (Tin House, 11/5)

The final installment of Tommy Picos Teebs tetrology (preceeded by IRL, Nature Poem, and Junk), this collection from the superstar queer Indigenous writer-hero returns us to the character of Teebs, a kind of alter-ego. Like his previous volumes, Feed is full of irreverent humor, razor-sharp stanzas, and stream-of-consciousness philosophy. Pico explores uncertainty and seasonality, the horror and ubiquity of todays headlines, friendship and dating, food and aliens, and of course, a few signature dick jokes. Nothing is quite like the experience of reading Picos workits a simultaneously heady and grounding adventure.

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13 Books by LGBTQ Writers to Read This Fall - Rewire.News

Recommendation and review posted by Bethany Smith

This story from the Australian Aviation archives comes from October 2012, when Ellis Taylor wrote about the Bombardier Dash 8 operations in Australia.

A Skippers Dash 8-300. (Paul Sadler)

Since the late 1980s, the Bombardier Dash8 has been a staple of regional aviation in Australia, with various iterations of the turboprop in service across the country.

Today, the 84 Dash 8 aircraft registered in the country serve a number of regional communities, both on scheduled services and in charters, while some have also been modified for special missions.

Those form part of a global fleet of over 1,000 Dash 8s of all types which fly in practically every corner of the globe.

Development of the aircraft started in the late 1970s when De Havilland Canada (DHC) found there to be large demand for a 30-40 seat commuter aircraft. Part of that requirement was to replace the venerable Fokker F27 Friendship, while it also provided a step up from smaller 19seat regional aircraft such as the Fairchild Metroliner.

It also followed the technically successful but sales-challenged Dash 7. That aircraft, which featured four turboprops and a capacity for 50 passengers, was legendary for its short takeoff and landing (STOL) abilities, capable of operating from airstrips as small as 610m.

At the time, it was envisioned that networks of STOL-ports could be established across Europe and North America close to major city centres.

However, the Dash 7 was a relative sales flop, with only 113 produced. While the STOL capabilities were impressive, many commuter airlines in North America and other markets simply did not require it. The higher fuel and maintenance costs of the four engines also made it uneconomical against other types.

That led to the development of the Dash8-100, which made its first flight in 1983 and entered service in 1984.

Powered by two Pratt & Whitney Canada PW120 turboprops driving four-bladed propellers, the Dash 8 promised the capability of carrying up to 39 passengers in an airliner-style cabin. And although it did not have the STOL capability of the Dash7, the aircrafts high wing and still respectable landing performance made it well suited to the regional environment.

The aircraft quickly became a success in the market, with many sold to a range of operators around the world.

Over time, DHC also added tweaks to the 100 series Dash 8. In 1990, it introduced the -101 which had additional headroom in the cabin and introduced PW120A engines. This was followed in 1992 with the -106 variant which introduced more powerful PW121 engines capable of producing 1,605kW (2,150shp) of power, and improved the aircrafts climb and short field performance.

Following the early success of the Dash8 in the 37-seat category, DHC pressed ahead in 1986 with plans to stretch the basic airframe and create a variant capable of accommodating 50 passengers.

That resulted in the launch of the Dash8300, which featured a 3.43m stretch to the fuselage with a stretch to the basic airframe to accommodate 50 seats and more powerful PW123 turboprops.

The first Dash 8-300 made its first flight in May 1987, with first delivery in May 1989.

The underside of a Skippers Dash 8-300. (Les Bushell)

As with the -100 series before it, DHC would launch improved variants of the -300, utilising different engine combinations to deliver more performance. The -311 was introduced in 1990 with new PW123A engines and a revised interior, while the -300A allowed for increased payload.

Then in 1992 DHC announced plans to add further enhancements to the basic -100 design that became the -200 series. Using the same fuselage and wing design as the -100, this variant introduced PW123 engines as used on the -300 series, giving it better range and performance in hot and high conditions. The type entered service in 1995, and most of the 105 produced remain in service today.

A Skytrans Dash 8 Q300 at Sydney Airport in August 2013. (Rob Finlayson)

For a number of years, Dash 8 production and development continued on while DHC underwent a series of changes. In 1986 the company was sold by the Canadian government to Boeing, which later tried to use it to win a significant aircraft order from Air Canada. Following the loss of that competition to Airbus, Boeing immediately put the company up for sale, and in 1992 it was sold to transport conglomerate Bombardier.

Despite the changes in ownership during the late 1980s and early 1990s, DHC/Bombardier continued to work on further developments to the Dash 8. This included the addition of an ingenious active noise and vibration suppression system being added across the lineup in 1996, and resulted in the Q designations being added to aircraft after that.

In total, Bombardier delivered 299 Dash8-100s, 105 Q200s and 267 Q300s before production of those types ceased in 2009 due to the backlog for the models being filled, and production switching solely to the 74-seat Q400.

A file image of a QantasLink Q200 at Mt Hotham. (Australian Aviation archive)

Q400

As with the -300/Q300 before it, the Q400 was largely based on the basic Dash8 design but with a further stretch. That necessitated a number of modifications to the aircrafts tail and wings.

The other major change was to the engines, with the model introducing the PW150A engines capable of producing 3,781kW (5,071shp) and driving six-bladed Dowty composite propellers. These engines also introduced full authority digital engine control (FADEC) to the Dash 8.

In the cockpit, Bombardier added a Thales digital avionics suite, making the cockpit more aligned to modern airliners than the previous Dash 8 models.

The Q400 also introduced two entry doors at the forward and aft ends of the aircraft, while earlier Dash 8s only featured front access doors.

The aircraft had its first flight in January 1998, with Canadian certification awarded in 1999. The first delivery to launch customer SAS Commuter occurred in January 2000.

The Q400 is able to cruise at 360kt, putting it closer towards the speeds of regional jets. Added to that, the aircraft has a ceiling of 27,000ft, allowing it to fly higher than other regional turboprops.

Despite those capabilities, the Q400s reputation became tarnished in 2007 following a series of landing gear failures in Europe. Launch customer SAS even publicly stated that it would dispose of its fleet as a result of the accidents, but the airline later ordered more aircraft after reaching an agreement on compensation with Bombardier.

A file image of an Air Niugini Q400. (Australian Aviation archive)

The Q400 has also had some troubles locally with landing gear issues. Following a manufacturer-mandated inspection in August 2010, QantasLink was forced to ground five Q400s temporarily to replace certain parts of the landing gear after cracks were found.

While it has been the mainstay of production for a number of years, Bombardier has more recently transitioned across to solely produce the upgraded Q400 NextGen.

Compared to the standard Q400, the NextGen has an updated cabin with improved lighting, larger overhead bins and revised windows, while it has also delivered some improvements in maintenance costs and fuel burn.

Last year Bombardier announced that it would offer a two-class configuration on the Q400 NextGen which would also be offered as a retrofit for existing Q400 operators. United Express is the first carrier to operate a Q400 with the three-abreast business class configuration.

Despite the general turn back towards operating turboprops over regional jets, the Q400 has lagged in the sales race, with the rival ATR 72 stealing a number of orders.

On a global scale, the Q400s sales success has been somewhat muted until recently. A number of larger competitions saw orders go to the rival ATR 72, which some analysts say has a lower acquisition price and lower fuel burn.

Nevertheless, Bombardier has recently had significant orders from Canadian carrier WestJet and Polands EuroLot, which have helped to boost its backlog to 54 aircraft.

QantasLink Q400 named Darling Downs at Wellcamp Airport. (Wellcamp)

Local operations

With regional services such a large part of Australias aviation network, it was inevitable that the Dash 8 would have a long career flying in southern skies.

Eastern Australia Airlines received its first Dash 8-100 in 1988. That was followed in 1998 by the arrival of Sunstate Airlines first -200 series aircraft. Sunstate later took delivery of its first Q300 in January 2000, while the Q400 was added to its fleet from January 2006.

Today QantasLink operates 27 Q400s, 16 Q300s and five Dash 8-200s, with one Q400 still to be delivered. The aircraft now operate in almost all states, with routes ranging from Sydney-Canberra right up to the much longer Cairns-Port Moresby sector. The -200 series aircraft are also used to maintain the link between Sydney and Lord Howe Island.

A QantasLink Q300. (Australian Aviation archive)

QantasLink disposed of its last -100 series aircraft in 2010, with most of them joining the other major Dash 8 operators, Cairns-based Skytrans and Perth-based Skippers Aviation.

Skytrans now operates 11 Dash 8-100s and three Dash 8-300s, while Skippers has a fleet of four Dash 8-100s and six Dash8-300s.

Both operators primarily use the aircraft on mining charter services, where the aircrafts range and hot and high capabilities have been especially valued, as well as its ability to operate from unpaved airstrips when serving remote mine sites. The carriers also use them on their limited regular public transport (RPT) networks to remote centres to great effect.

For Skippers, adding the -300 to its fleet has allowed it to ride the resources boom, with the aircraft allowing for bigger shift changes at some of the mines it flies to. Similarly, the aircraft has also given it the option to up-gauge some of its RPT services in WA to meet demand.

Cobham Aviation Services has also been a longtime Dash 8 operator, having leased in -100 and -300 series aircraft to meet the needs of its resource charter clients.

Cobhams Surveillance Australia is also a notable Dash 8 operator, flying six Dash8-200s and four Dash 8-300s, which have been modified to carry a specialised sensor system and larger fuel tanks to enable it to carry out maritime surveillance missions on behalf of Customs.

A file image of a Dash 8 aircraft operated by Cobham conducting aerial surveillance. (Cobham Aviation Services)

Across the Tasman, now-defunct carriers Ansett New Zealand and Origin Pacific operated Dash 8-100s and -300s.

Air New Zealand also ordered Q300s to replace the Saab 340s operated by Air Nelson. The first one was delivered in Montreal in July 2005 and today the airline operates 23.

The Air New Zealand order was something of a coup for Bombardier at the time, especially as one of the carriers other regional subsidiaries, Mount Cook Airline, already operated ATR 72s.

Further north, Dash 8s have been long-term residents of Papua New Guinea, having been flown for many years by AirNiugini and Airlines PNG, providing vital internal connections and also flying between Port Moresby and Cairns.

Air New Zealand Q300 ZK-NEP at Wellington. (Gary Hollier)

Future Developments

Despite the earliest model Dash 8s now having been out of production for three years, demand for the -100, -200 and -300 series remains high and overall plenty of life remains in the fleet, particularly with the manufacturer continuing to support the oldest aircraft still flying.

Bombardier announced in 2009 that it would extend the life of the -100 series out to 120,000 cycles from the previous 80,000 cycles through an Extended Service Program.

The company says that on current usage trends the effective life of the aircraft could be extended for up to 12 years.

One operator Australian Aviation spoke to said that the life extension had meant that the values of Dash 8-100s had increased and that it was getting hard to find aircraft available. As a result, some operators with a requirement for more aircraft are moving up to the larger -300 which are easier to source on the global market.

Another problem for Dash 8 operators has been the lack of new aircraft in the market, especially in the 37-seat category.

The Q400 introduced a Thales digital avionics suite. (Bombardier)

Further support is coming from Fokker Services, which last year signed an agreement with Bombardier to provide spares and logistical services to Dash 8 operators. The service builds on its ABACUS program which helps to support legacy Fokker types.

At the same time, Bombardier has also given every indication that it plans to further extend the Dash 8 design up into a new 90-seat aircraft in future.

In 2007 Bombardier started talking of a Q400X, which would use the basic airframe but feature a stronger wing and upgraded engines and would be aimed at operating on short but dense regional routes. Originally, the manufacturer had indicated that the Q400X could be developed to enter service as early as 2013.

The Q400X talk has also prompted ATR to closely examine a larger aircraft, with the two manufacturers playing a game of cat and mouse over the last few years.

More recently engine manufacturers General Electric and Pratt & Whitney have indicated that they are working on developing more powerful turboprops for both Bombardier and ATR.

However, this year Bombardier has cooled on attacking the 90-seat market, with recent comments from its marketing team indicating that any move on that segment would likely come after 2016.

In the meantime though, the Dash 8 looks set to continue its legacy in Australia of being a flexible and hardy regional workhorse.

VIDEO: A look at a QantasLink Dash 8 Q200 taking off from Lord Howe Island Airport from the wiiwheel64 YouTube channel, which highlights the excellent field performance of the aircraft.

Postscript: In November 2018, Bombardier announced the sale of the Q400 program, as well as all assets, intellectual property and type certificates associated with the out-of-production Dash 8 series, to Longview Aviation Capital. It established a new company with a famous name De Havilland Aircraft of Canada Ltd for manufacturing the Q400 turboprop.

This story first appeared in the October 2012 edition of Australian Aviation. To read more stories like this, subscribe here.

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From the archives: Regional workhorse: Bombardier Dash 8 in Australia - Australian Aviation

Recommendation and review posted by Bethany Smith

From the time of the Babylonian Exile, Jews have been spread far and wide, carrying with us mementos of our ancient past in our blood, spit and the microscopic double helix of our DNA. Among the best tools the Chosen People have for finding a link to antiquity is bleeding-edge technology that analyzes our genes and while that tech is new, its largely confirmed a familiar story of shared roots in the Middle East.

Through advancements in mapping the human genome and the study of traditionally diseases like Tay-Sachs, scientists and historians are closer than ever before to learning where the Jewish people originated, and where we ended up.

The science involved in genetic study of Jews has advanced immensely since its early days, when physical anthropologists Joseph Jacobs and Maurice Fishberg studied outward markers like stature, head size and pigmentation, said Harry Ostrer, the director of Genetic and Genomic Testing at Montefiore Medical Center and the author of Legacy: A Genetic History of the Jewish People (2012). Working in the late 19th and early 20th Century, when Fishberg published The Jews: A Study of Race and Environment (1911), his and Jacobs primitive research which had a troubling analogue in the measurements used by Nazi race science was surpassed by a new effort that emerged, fittingly, in Mandatory Palestine.

After moving to Palestine in the 1930s, Chaim Sheba and other physician investigators observed that the Jewish communities flocking to the land from across the globe did not much resemble each other. Sheba and colleagues believed these differences ought to be studied. This work culminated in 1961, when Sheba and geneticist Elisabeth Goldschmidt held the Conference on Human Population Genetics in Jerusalem.

By the 1970s, studies using blood groups conducted by researchers like Arthur Mourant and Batsheva Bonne-Tamir advanced our understanding of Jewish common ancestry, as did studies in the 1990s focused on matrilineal mitochondrial DNA and patrilineal Y chromosomes. But it wasnt until Jewish genomes, an individuals entire genetic code, started being analyzed in the 2000s, following the breakthroughs of the Human Genome Projects push to map all human genes, that this knowledge became less abstract.

Before genome-wide analyses, There was a real lack of precision, Ostrer told the Forward over the phone. The Y chromosomal and mitochondrial studies supported the idea, for instance, for European Jews, that they had both Middle Eastern and European origins. But it just wasnt with the degree of accuracy or precision that happened when the genome-wide studies were done.

Ostrers initiative, the Jewish HapMap Project, is one of a number of studies propelled by advancements in genomic technology, which can now determine the order 20,000 genes (and 3 billion nucleotide base pairs) at one time in around one to two months. In recent years, these efforts have yielded insights into the inter-connectedness of the Jewish people.

Part of why that research is so fascinating is its telling us about aspects of Jewish history that are not recorded in texts or reflected in archaeology, said Steven Weitzman, the Ella Darivoff director of the Katz Center of Advanced Judaic Studies at the University of Pennsylvania and the author of The Origin of the Jews: The Quest for Roots in a Rootless Age (2017).

Weitzman, who collaborated with Stanford geneticists in his research, noted a few early landmark studies that seemed to confirm traditional narratives, that link Jewish Diasporic communities to common tribal ancestors in modern day Israel. One of the earliest, from 1997, was a Y-chromosomal analysis of Kohanim, which indicated a common male ancestor some 3,000 years ago.

That doesnt mean that ancestor is Aaron as Jewish tradition would indicate and its since been challenged but it was really striking that they could see reflected in the genetic code of these people that they did share a common male ancestor somewhere thousands of years ago.

Since the completion of the Human Genome Project in 2003, more ambitious studies have been mounted, Weitzman said, including Doron Behars 2010 attempt to draw a genome-wide map of the Jewish people a goal shared by Ostrers HapMap Project by sampling DNA from 14 global communities. The study found that many of the communities differed genetically from their non-Jewish neighbors and had roots in the Middle East. While that tracks with conventional history, other research, such as a 2013 study by Martin B. Richards, concluded that the matrilineal line of Ashkenazic Jewish descent has its roots in native European women. Behar, who, with Karl Skorecki, had concluded years earlier that 40% of Ashkenazi Jews can trace their origins to four women of Middle Eastern extraction, disagreedwith this assessment.

[Genetics] doesnt necessarily corroborate the traditional Jewish understanding, Weitzman said, but it often does. And thats why some people are a little skeptical. Its been reaffirming a very conservative understanding of Jewish history.

While the majority of scientists and historians support the view that Jews originated in the Middle East, there are vocal skeptics, notably Israeli geneticist Eran Elhaik and Israeli historian Shlomo Sand. Sand and Elhaik subscribe to the Khazar Hypothesis, popularized by author Arthur Koestler, which argues that Ashkenazi Jews are descended from a Turkic people called the Khazars, who converted. Ostrer and the broader scientific community have dismissed this theory, and Elhaiks specific research, as agenda-driven science. Weitzman agrees that the theory presents a weak argument and has been used to discredit Jews and Zionism by distancing Ashkenazim from claims to a Middle Eastern homeland.

The preponderance of evidence, based on a number of studies, indicates a conventional view of Jewish diaspora, Ostrer claims. Jews appear to have migrated east along the Silk Road and across the Arabian Sea and westward into Europe and Northern Africa by way of the Greek and Roman empires. These migrants intermingled with locals sometimes converting them to the faith along the way. Despite this dispersion, Jews remained largely endogamous, marrying within our ethnic group, relative to native populations.

The same endogamy has historically resulted in genetic diseases, which, by medical necessity, have made Jewish genes among the most studied. While Ostrer cautions that some consumer-available genetic tests can be deceptive, appearing to indicate closer kinship between Jews who share deep ancestral lines, he maintains that the testing for potential disease carriers has improved tremendously with companies like JScreen, Invitae and Myriad.

Currently, those with Sephardic and Mizrahi backgrounds (roughly 10% of American Jews) dont have the advantage of seeing that heritage listed in most popular genetic testing service results. But these populations are sufficiently studied by geneticists and often analysis of these groups can lead to interesting places. Recently, disease mutations like BRCA 1 and 2 and hereditary ovarian and breast cancer typically found in Jews have been surfacing among Latino populations in the United States.

In a 2011 paper, Ostrer and his colleague Christopher Velez argued that these genetic predispositions were carried to the New World by Converso Jews in the late 15th century. New research from 2018 appeared to confirm this, finding that a wide swath of Latin Americans have ancestry from newly-converted Christians.

Overall, DNA testing and studies have enriched the Jewish historical record beyond potsherds and texts dating back to the Greek and Roman period. But while these science-based tools are often less ambiguous than previous historical documents, they have not been embraced by all historians. The reason, ironically enough, is recent history.

Theres a lot of resistance to [genetic research] within the field of Jewish studies, Weitzman said. A lot of people remember or have in mind the role of race science in Nazism. So the idea that Jewish scholars would look in any way to genetics to understand Jewish identity or Jewish history and origins can make people concerned.

PJ Grisar is the Forwards culture fellow. He can be reached at Grisar@Forward.com.

This story "How Genetics Paint A Picture Of The Jewish Past" was written by PJ Grisar.

Continued here:
How Genetics Paint A Picture Of The Jewish Past - Forward

Recommendation and review posted by Bethany Smith

The mechanism isnt clear but abnormalities on the Y chromosome are in the frame

Poor sperm function (male infertility) causes nearly half of all infertility. The observation that poor sperm function is commonly associated with developmental genitourinary defects such as cryptorchidism suggests that male infertility could be a risk marker for later disease. Unfortunately, we know little about the natural history of male infertility beyond reproductive life, owing to historical social stigma experienced by affected men. Studies shedding light on the future health of infertile men should be welcomed, and in the linked paper (doi:10.1136/bmj.l5214) Al-Jebari and colleagues report analyses of registry data from the whole population of Sweden over 20 years. Their results provide the strongest evidence to date that risk of prostate cancer may be increased in infertile men. Importantly though, a causal relation cannot be assumed.1

Al-Jebari and colleagues included 1.2 million men in Sweden who fathered a first born child during the study period, either spontaneously (97% of men) or following one of the two leading assisted reproductive technologiesintra-cytoplasmic sperm injection (ICSI) and in vitro fertilisation (IVF). The study found that men becoming fathers through IVF and ICSI had a significantly higher risk of prostate cancer than men who fathered children naturally. The risk of early onset prostate cancer (diagnosed before age 55 years) was particularly high for men fathering children through ICSI, a technology used for men with the most severe forms of infertility.

The use of ICSI and IVF as proxies for poor sperm function is reasonable in a population study of this size, although it has obvious limitations; IVF is often given to couples with infertile female partners and male partners with entirely normal sperm function. Men who had assisted reproduction were older and educated for longer than fathers conceiving naturally. In view of that, the researchers adjusted for both paternal age and education level in their analysis. Furthermore, to prevent bias, men with previous cancer or testosterone replacement were excluded.

Previous studies have also observed an association between male infertility and subsequent prostate cancer.23 A population based study of 22562 infertile men reported that men with male infertility were 2.6 times more likely to develop a high grade prostate cancer than were their age matched controls.3 More recent observational research found a correlation between lower semen quality and worse scores on an index of general health. Cancer diagnoses were included in the index.4 The evidence is not entirely consistent, however. Some studies do not support an association between infertility and risk of prostate cancer.56 Others have concluded that men never fathering children actually have a reduced risk compared with men with at least one child,78910 although the absence of children is a poor surrogate for infertility.

These authors acknowledge the limitations of their study.1 Firstly, the study did not include infertile men who were unable to father children. These men might be expected to have a higher risk of prostate cancer than infertile men who managed to father children. Secondly, the mean age at follow-up was 45 years, so these findings are unlikely to quantify risk of prostate cancer over a lifetime. Lastly, the incidence of prostate specific antigen testing in population groups might have provided direct evidence that infertile men were not subjected to enhanced cancer screening.

How male infertility could be linked biologically to risk of prostate cancer is not yet clear. Possibilities include a genetic association between microdeletions in the Y chromosome, which are known to cause severe male infertility, and genes on the same chromosome known to be associated with prostate cancer.11 Mutations in DNA repair genes and epigenetic and environmental modulators have also been suggested to link male infertility and prostate cancer.1213

Middle aged men are often interested in their risk of prostate cancer; age, family history, and Afro-Caribbean ethnicity are major risk factors. However, screening is controversial owing to lack of survival benefit and the harms from overdiagnosis and overtreatment that can follow a positive screening test.14 In the absence of a plausible mechanism of action or proof of causation, justifying screening for prostate cancer in all infertile men is difficult. However, further research on the possible future complications of male infertility would be welcomed by patients and will help clinicians to counsel all infertile men about their future health.

We are grateful to our patient representative, James Tyerman, for providing valuable comments on the manuscript.

Visit link:
Male infertility linked to risk of prostate cancer - The BMJ

Recommendation and review posted by Bethany Smith

Zambia, a country in southeast Africa, has approximately 1,200 lions, one of the largest lion populations on the continent. More than 40% of the U-shaped country is protected land, with over 120,000 square miles of national parks, sanctuaries and game management areas for lions to roam.

Zambian lions are split into two subpopulations, with one in the Greater Kafue Ecosystem in the west and the other in the Luangwa Valley Ecosystem in the east. Between these two geographically different regions lies Lusaka, Zambias largest city, which is surrounded by farmland.

People had assumed that the two groups of lions did not even could not mix. After all, theyre separated by a geographical barrier: the two regions feature different habitats, with the east an offshoot of the Great Rift Valley system and the west part of the southern savannas. The lions are also separated by whats called an anthropogenic barrier: a big city that lacks wildlife protection, making it seemingly unsuitable for lions.

So my colleagues and I were surprised when we found that a small number of lions are in fact moving across the area in between presumed to be uninhabitable by lions. These sneaky lions and their mating habits are causing the high levels of genetic diversity we found in the entire Zambian lion population.

Working with the Zambian Wildlife Authority, biologist Paula White collected hundreds of biological samples from lions across Zambia between 2004 and 2012. Eventually a box of this hair, skin, bone and tissue, meticulously packaged and labeled with collection notes and sampling locations, arrived at my lab at Texas A&M University.

Our goal was to investigate genetic diversity and the movement of various genes across Zambia by extracting and analyzing DNA from the lion samples.

From 409 lions found inside and outside of protected lands, I looked at two kinds of genes, mitochondrial and nuclear. You inherit mitochondrial DNA only from your mom, while you inherit nuclear DNA from both of your parents. Because of these differences, mitochondrial and nuclear genes can tell different genetic stories that, when combined, paint a more complete picture of how a population behaves.

My mitochondrial analysis verified that, genetically, there are two isolated subpopulations of lions in Zambia, one in the east and one in the west. However, by also looking at the nuclear genes, we found evidence that small numbers of lions are moving across the unsuitable habitat. Including nuclear genes provided a more complex picture that tells us not only which lions were moving but also where.

The amount of variation from alternate forms of genes found within a population is known as genetic diversity. Genetic diversity is important for a wildlife population because more genetic options give animals a greater chance for adaptation in a changing environment. Genetic diversity can also tell biologists about ways a population can fluctuate.

To a geneticist, migration, also referred to as gene flow, is the movement of genes from one geographical place to another. Mitochondrial DNA, inherited from the mother, can only tell researchers where genes from mom have been.

In the lion mating system, males travel long distances to find new prides, while females remain in or close to the pride they were born in. So, for the lion, its primarily males that are responsible for the movement of genes between prides. This male-mediated gene flow explains the lack of gene flow seen in mitochondrial genes compared to that of nuclear genes female lions arent making the journey, but they do mate with new males who come from far away.

Male-mediated gene flow has helped keep the lions of Zambia genetically healthy, increasing genetic diversity by introducing new genes to new areas as male lions move between subpopulations. The eastern and western subpopulations each have high levels of genetic diversity; since only a few lions move between the groups each generation, the subpopulations stay genetically distinct.

My colleagues and I were also able to determine where the lions are moving based on which individuals are more genetically similar to each other. Lions in the North and South Luangwa National Parks, part of the eastern subpopulation, appear completely separated from the western subpopulation. Gene flow is occurring through the southern regions of the eastern subpopulation.

Lions are most likely traveling a route between the Lower Zambezi National Park and eastern corridor to the Kafue National Park in the west, possibly along the Kafue River. We cant tell which way theyre moving, but by looking at where lions are more closely related, we can see where genes are being moved.

Human-lion conflict is a big issue in Zambia, particularly outside of protected land. If lions were moving across human dominated areas, youd think theyd be seen and reported. But these lions are sneaking through virtually undetected until we look at their genes.

As a large, charismatic carnivore, lion research and conservation influences many other species that share their habitat.

Wildlife managers can use these findings to help with lion conservation and other wildlife management in and around Zambia. Now that we generally know where lions are moving, managers can focus on these areas to find the actual route the big cats are taking and work to maintain or even increase how many lions can move across these areas. One of the ways of doing this is by creating more protected land, like corridors, to better connect suitable habitat.

[ Youre smart and curious about the world. So are The Conversations authors and editors. You can read us daily by subscribing to our newsletter. ]

Link:
Sneaky lions in Zambia are moving across areas thought uninhabitable for them - The Conversation - US

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Images of child sex abuse have reached a crisis point on the internet, spreading at unprecedented rates in part because tech platforms and law enforcement agencies have failed to keep pace with the problem. But less is understood about the issue underlying it all: What drives people to sexually abuse children?

Science in recent years has begun to provide some answers. One thing most pedophiles have in common: They discover, usually as teenagers, that their sexual preferences have not matured like everyone elses. Most get stuck on the same-age boys or girls who first attracted them at the start of puberty, though some retain interest in far younger children.

People dont choose what arouses them they discover it, said Dr. Fred Berlin, director of the Johns Hopkins Sex and Gender Clinic. No one grows up wanting to be a pedophile.

[Read The New York Timess investigation into the spread of online child sex abuse.]

Over the past generation, psychologists, forensic specialists and others have studied pedophilia, a disorder characterized by recurrent, intense arousing fantasies, urges or behaviors involving sexual activity with a prepubescent child, according to psychiatrys diagnostic manual. These experts have interviewed patients in depth, piecing together life histories and performing a variety of psychological and anatomical measures.

While no study offers a complete picture, a portrait is emerging one that helps elucidate the mental dynamic behind the surge in abuse images and the deepening depravity they depict. These findings also defy common stereotypes about what pedophilia is, and what the risks are for engaging in physical abuse.

A majority of convicted offenders are men who prey on children ages 6 to 17. But women also commit hands-on offenses; rough estimates put the rate of pedophilic attraction at 1 to 4 percent in both men and women. Studies suggest that a small subset of male and female pedophiles have an interest in toddlers, or even infants.

As scientists seek to understand how the disorder develops, there is growing consensus that the origin is largely biological. This view is based in part on studies pointing to subtle physical traits that have a higher incidence among pedophiles.

The biological clues attached to pedophilia demonstrate that its roots are prenatal, said James Cantor, director of the Toronto Sexuality Center. These are not genetic; they can be traced to specific periods of development in the womb.

Psychological and environmental factors may also contribute, though it is not yet clear what those are or how they interact with developmental conditions.

By contrast, the common presumption that pedophiles were themselves abused as children now has less support. Child victims are at far greater risk of future substance abuse, depression, persistent traumatic stress or criminal aggression than of becoming molesters. The vast majority of offenders deny any sex abuse in their childhood, even though they could garner sympathy in court by doing so, experts say. A chaotic childhood increases the likelihood of a chaotic adulthood, of any kind, Dr. Cantor said.

The relationship between viewing or collecting images and committing hands-on abuse is a matter of continuing debate among some experts, and one that is critical to evaluating the risk an offender poses. Until recently, the prevailing view was that only a minority of people caught viewing such images, between 5 and 20 percent, also committed physical abuse.

That perception began to change in 2007, when a pair of psychologists at the Federal Bureau of Prisons reported that 85 percent of convicted online offenders acknowledged in therapy that they had raped or otherwise sexually abused children.

That finding circulated widely before the study was formally published, creating an uproar among therapists, researchers and law enforcement specialists. The prisons bureau balked at publishing it at all, and withdrew it from a peer-reviewed journal close to its release date.

Many cited concerns that the study sample was biased: It was based on the confessions of 155 convicts who had sought out therapy in prison, not on a representative sample of pedophiles, a much broader group with diverse habits.

It was what we call a convenience sample that was a legitimate criticism, said Michael L. Bourke, a co-author of the study with Andres E. Hernandez, in a telephone interview. Dr. Bourke is now chief of the behavioral analysis unit of the United States Marshals.

Since then, several other studies have supported the prison finding, if not precisely the 85 percent number. In one, inspectors from an array of government agencies interviewed 127 online offenders shortly after their arrests. Less than 5 percent admitted to previously molesting at least one child.

When agents followed up with more in-depth, polygraph-assisted methods, another 53 percent admitted to hands-on offenses, for a total of nearly 60 percent.

This was not a convenience sample; these were offenders, some of whom had downloaded just a single image, with no known history, from all over the country, interviewed by people from different agencies, Dr. Bourke said. They had zero incentive to admit to a previous offense very much the opposite.

The high rate of previous, hands-on offending undermines another common assumption about pedophiles. We shouldnt assume that viewing online images leads to abuse of a child victim in person, said Joe Sullivan, a specialist in sex crimes against children in Ireland and Britain. In my clinical experience, its the other way around. Most of these men have already committed hands-on offenses.

From this point of view, downloading abuse images and especially connecting with groups of like-minded pedophiles online does not awaken latent desires. The desires are very much awake and, in many cases, have already been acted on. But the images and online communities can help erode inhibitions further, drawing pedophiles into more frequent or more aggressive acts, Dr. Bourke said.

What you see, in their search histories, he said, is that they learn how to evade law enforcement, they become more confident and they begin to use cognitive distortions to overcome their moral inhibitions.

Some therapists and researchers say these findings from law enforcement threaten to unfairly tar people who never act on their desires. This group certainly exists theyre sometimes called virtuous pedophiles but in an era of increasing alarm over the proliferation of online abuse, they are going only further underground.

That is a shame, a tragedy, Dr. Cantor said. That is the group we need to learn about. Thats the kind of person wed like our clients to become, a person whos aware of the urges and learns to effectively manage them.

Learning to manage a drive as visceral, and often consuming, as sexual desire is possible, therapists say, but it cannot be shut off; nor can it be replaced, the way heroin can be swapped for methadone. Treatment can require drugs that reduce circulating testosterone and software that limits online browsing habits.

Often, therapy addresses substance abuse as well. Studies suggest that at least 40 percent of sex offenders were using drugs or alcohol when they committed their crimes.

The important thing, I think, is that people know that treatment is possible, Dr. Berlin said. Theres a subgroup out there, they refer themselves here, and they are quite convinced that they do not want real-life sex with children.

Original post:
Preying on Children: The Emerging Psychology of Pedophiles - The New York Times

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Like the deserts of the Antarctic, or the deepest parts of the sea, Mono Lake in California is an inhospitable place for most life forms. Apart from bacteria and algae, it appears only brine shrimp and diving flies can put up with its super-salty waters.

But there's more to this body of water than meets the eye. Researchers at the California Institute of Technology have recently discovered eight more species of microscopic worm thriving in and around the lake, and one of them is a brand new kind of weird.

One of the newly-discovered species of nematode - for now called Auanema sp. - has not one, not two, but three different sexes, the team reports, and it can survive a dose of arsenic 500 times what is humanly possible.

When it comes to differentiation of the sexes, nematode species usually keep it simple, dividing into hermaphrodites and males. But Auanema sp. also has worms of the female sex. Furthermore, they have other interesting sex characteristics, as the researchers note "the arrangement of genital papillae in Auanema sp. males is unique in the genus."

As if that's not radical enough, the team says this microscopic worm also gives birth to live offspring, a unique approach in the typically egg-laying nematode world.

It's an extreme creature in an extreme place, and that's probably not a coincidence. The team thinks this worm's strange characteristics are part of what keeps it alive in the hyper-salty, alkaline waters of Mono Lake.

"Extremophiles can teach us so much about innovative strategies for dealing with stress," says Pei-Yin Shih.

"Our study shows we still have much to learn about how these 1,000-celled animals have mastered survival in extreme environments."

Comparing the strange new species of nematode to others in the same genus, the researchers found a similarly high arsenic resistance among two sister species.

And yet the curious thing was, none of these creatures actually lived in environments with high arsenic levels. There had to be another reason for this astonishing tolerance.

"Previous Auanema species were isolated from rich soils and dung, which can contain high concentrations of phosphate," the authors suggest.

"Since arsenic uptake occurs adventitiously via phosphate transporters, it is conceivable that adaptation to high levels of phosphate in the environment could lead to increased arsenic resistance as well."

In other words, nematodes might be pre-adapted to life as an extremophile. They could have a genetic resiliency and flexibility that makes it easier for them to live in harsh places like Mono Lake.

Before this study, only two other species had been found in this lake - which is three times as salty as the ocean and has an alkaline pH greater than baking soda. Yet even still, the discovery of eight more species wasn't all that surprising to researchers.

Nematodes are the most abundant type of animal on the planet, so even in the harsh environment of Mono Lake, there's a good chance you'll find them.

Over the course of two years, researchers from Caltech isolated nematodes from across the lake and found multiple niches in which these nematodes were thriving. And their numbers included microbe grazers, parasites and predators.

"Thus, nematodes are the dominant animals of Mono Lake in species richness," the authors conclude.

"Phylogenetic analysis suggests that the nematodes originated from multiple colonisation events, which is striking, given the young history of extreme conditions at Mono Lake."

To say these creatures are opportunistic is an understatement. For every human on Earth, there are roughly 57 billion nematodes, and in barely no time at all, these creatures can set up shop in some of the most extreme places on Earth.

"It's just so cool finding another place where there are lots of nematodes and not much else," geoscientist Tullis Onstott who was not involved in the study told The Scientist.

And who knows, maybe when there's not much else, there will still be nematodes.

The findings were published in Current Biology.

The rest is here:
A Worm With Three Sexes Has Been Discovered Thriving in a Nearly Lifeless Lake - ScienceAlert

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Fans greeted the news that Sam Neill, Laura Dern and Jeff Goldblum will join the cast of Jurassic World 3 with a roar of approval. Is this a shot in the arm the franchise needs after the relative disappointment of Fallen Kingdom, or is this Universal shamelessly pandering to nostalgia?

Maybe its both. The return of the beloved principals of the first Jurassic Park is certainly welcome, but how effective that return is depends on how well they are used.

The first Jurassic World in 2015 was a huge success, making $652 million here and $1.7 billion worldwide, making it the franchise leader before you adjust for inflation. Although some critics knocked it for being sexist and too derivative of Jurassic Park, it was generally well reviewed, with 72% on Rotten Tomatoes.

Fallen Kingdom came out three yearslater, and something was missing enthusiasm. The sequel made $417 million here, a disappointment somewhat salvaged by making $1.3 billion worldwide, but the reviews hit a franchise low of 48%. Fans and critics alike noted the dour tone, with a structure that seemed too close to The Lost World. Both the 1997 and 2018 contrived reasons to return to dinosaur islands failed to capture the wonder of their predecessors. Both movies also unleashed the dinosaurs on the mainland.

One aspect of the sequel that burned some fans was that the trailers highlighted an appearance by Goldblums Ian Malcolm, but he turned out only to have a cameo, with almost his entire performance contained in the trailer. One hopes Universal isnt making the same mistake again with Dern and Neill, although it sounds like all three will be more central to the story this time.

Jurassic World 3, due out in June 2021, will deal with the aftermath of the dinosaurs reaching the mainland. The Lost Worlds third act dealt with this too, but it cleaned up the mess relatively quickly. That movie had one dinosaur stomping around, while this one will have a whole bunch.

Colin Trevorrow, returning to the directors chair after only co-writing Fallen Kingdom, told EW the storywill be focused storytelling with dinosaurs all over the world. We really wanted this technology, this genetic power, to go open-source at the end of the film. What were suggesting is not just that these specific animals that we care about that were in captivity were freed, but also that the ability to create these animals has gone a little bit wider than our friend Dr. Wu. The open-sourcing of any technology, like nuclear power, thats the scary side for me.

In other words, it sounds like we get not only your traditional T-Rexs and velociraptors, but your mutant hybrid dinosaurs that caused so much trouble in the previous movies. Those greedy humans just never learn, do they? Chris Pratt and Bryce Dallas Howard return as well.

Many fans were delighted to see Neill, Dern and Goldblum back in the mix, with one person on Instagram saying, BEST DAY EVER! YOU GUYS KNOW HOW TO MAKE HAPPY THE FANDOM! A more cynical commenter on EW said, The original title of this story was: Universal resorts to Disney tactics by playing on Gen X nostalgia. (and Millennials disdain for watching ancient 90s movies.)

It almost goes without saying that Jurassic World 3 will make a ton of money. One hopes, however, that Universal isnt just handing out the rose-colored glasses of nostalgia. Sam Neill and Laura Dern came back for Jurassic Park III, and that movie in some corners is like the forgotten stepchild, so theres no guarantee their return will get the franchise back on course.

To be fair, dinosaurs roaming the mainland isnt something the franchise has played with for an entire movie, and if it fulfills the promise of the short film that came out, then Jurassic World 3 will make a good capper for the franchise provided they stop there.

See the original post here:
'Jurassic World 3': The Real Reason the Movie Needs Original Characters to Return - Showbiz Cheat Sheet

Recommendation and review posted by Bethany Smith

Global Male Hypogonadism Market: Snapshot

Hypogonadism in males refers to a condition in the male body where the testes show a significantly reduced level of functioning than normal. The overall result of male hypogonadism is a reduction in the rate of biosynthesis of male sex hormones. This state is more commonly known as interrupted stage 1 puberty. Hypoandrogenism, or the low androgen or testosterone level in a male can vary in severity from person to person. It is often the cause of partial or complete infertility. There are multiple forms of male hypogonadism and even more ways to classify them. Most endocrinologists commonly classify male hypogonadism on the basis of the level of defectiveness of the male reproductive system.

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In many cases, doctors also measure the level of gonadotropins to classify a patient between primary and secondary male hypogonadism. Primary male hypogonadism refers to the cause of the condition being due to defective gonads. There are different types of primary male hypogonadism, including Turner syndrome and Klinefelter syndrome. Secondary male hypogonadism is caused by defects in pituitary or hypothalamic glands. They include Kallmann syndrome and hypopituitarism.

Global Male Hypogonadism Market: Overview

Male Hypogonadism refers to a clinical condition, wherein the testes fail to produce enough testosterone leading to delayed puberty or incomplete development. The condition is related to impaired development of muscle mass, development of breast tissues, impaired body hair growth, and lack of deepening of the voice.

The male Hypogonadism market can be segmented by therapy, type, drug delivery, and geography.

The report presents an in-depth analysis of the global male hypogonadism market with current trends and future estimates to explain the imminent investment pockets. The quantitative analysis of the market for the forecast period from 2017 to 2025 will enable stakeholders to capitalize on the prevailing growth opportunities.

Global Male Hypogonadism Market: Trends and Opportunities

The top driver of the male hypogonadism market includes rising prevalence of testosterone deficiency among men, increasing infertility rates, and increasing awareness among individuals about hypogonadism treatment due to awareness drives organized by several governments across the world. Moreover, high risk of hypogonadism among the geriatric population with obesity and diabetes, and increasing prevalence of chronic disorders among the geriatrics are further expected to boost the markets growth.

However, factors such as high side effects of testosterone products are challenging the growth of testosterone replacement therapy market. Top players in the market are focused on research and development to introduce newer products with fewer or negligible side effects and improved results. For example, LPCN 1111, a product which is under development from Lipocine Inc., is a newer testosterone prodrug that utilizes Lipral technology for enhanced systemic absorption and for enhanced solubility of testosterone. Nevertheless, technological advancements are anticipated to extend new opportunities to the markets growth.

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Global Male Hypogonadism Market: Regional Overview

The global male Hypogonadism market can be analyzed with respect to the regional segments of North America, Asia Pacific, Europe, Latin America, and the Middle East and Africa. North America held the majority share of the global market in the recent past and is expected to retain its dominant position in the near future. This is mainly due to the rise in the number of individuals suffering from primary and secondary conditions of hypogonadism, and rising awareness among individuals about treatment options for the condition. Moreover, the presence of ultra-modern healthcare infrastructure and increasing popularity of technologically advanced products are expected to offer new opportunities for top players in this market. The region is closely followed by Europe.

Asia Pacific is expected to offer lucrative opportunities to this market due to the modernization of the healthcare infrastructure in the emerging economies of India and China and the increasing awareness about the treatment for the condition. In Asia Pacific, the increasing prevalence of hypogonadism and infertility rates along with the rising geriatric population base with diabetes and obesity are propelling the growth of this market. China, Taiwan, and Malaysia are some of the countries that display the highest rate of male hypogonadism.

Major Companies Mentioned in Report

Some of the key players in the male Hypogonadism market include AbbVie Inc., Astrazeneca plc, Eli Lilly and Company Ltd., Merck & Co. Inc., SA, Finox Biotech, Laboratories Genevrier, Teva Pharmaceutical Industries Ltd., Allergan plc, Bayer AG, Endo International plc, IBSA Institut Biochimque, and Ferring.

Key players are focused on product approval for growth considerations and to cater to the changing demand of the industry. The introduction of innovative and technologically advanced products is also the focus of key players to increase their market share and for serving patients in a better manner.

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TMR Research is a premier provider of customized market research and consulting services to business entities keen on succeeding in todays supercharged economic climate. Armed with an experienced, dedicated, and dynamic team of analysts, we are redefining the way our clients conduct business by providing them with authoritative and trusted research studies in tune with the latest methodologies and market trends.

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Male Hypogonadism Market Growth Opportunities and Dynamic Business by 2025 - NewsStoner

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Heart attacks can cause immediate death. But in survivors, the blockage of blood flow can kill so many heart muscle cells that heart failure can follow months or years afterwards. Heart disease is the leading cause of hospital admission and death in the United States.

A heart attack causes a loss of muscle and leaves the heart with a scar that does not contract and so impairs the hearts pumping function, says Tim Kamp, a professor of medicine who is co-leader of a new grant designed to attack two roadblocks that have stymied efforts to restore heart muscle with muscle cells grown from stem cells.

Kamp, who directs the Stem Cell and Regenerative Medicine Center at the University of WisconsinMadison, says, Everybody involved in treating these patients knows that this scarring often leads to a continual decline in heart function with heart failure and even death.

The UWMadison researchers used approved surgical devices to locate the damaged heart muscle, and then injected the supportive matrix and committed cardiac muscle cells. The circle outlines target zone established before surgery; black dots show the sites that were injected in this mouse study. Amish Raval, work performed at UWMadison in collaboration with Biologics Delivery Systems.

Sixteen percent of men, and 22 percent of women, develop heart failure after myocardial infarction heart attack. Coronary artery disease the category that includes stoppage of blood flow causes one in seven deaths in the United States.

Adult stem cell injections seemed a logical way to form new heart muscle cells and repair the damaged muscle. But in dozens of experiments, the cells either washed out of the heart or failed to develop into the specialized muscle cells the cardiomyocytes that power cardiac contractions. The benefits were mixed, modest at best, says Kamp.

After years of preliminary investigations, however, Kamp and Amish Raval, a professor of cardiology, researcher and entrepreneur, hope that a combination of two cutting-edge approaches would use a fabric-like material to prevent wash-out and successfully implant cardiomyocytes to damaged hearts.

Aided by a Regenerative Medicine Innovation Project grant from the National Heart, Lung, and Blood Institute, part of the National Institutes of Health, the two will lead a group to test that idea in pigs over two years.

Having committed cells could be a major advance, Raval says. The first stem-cells therapies started with cells that I call the model T. Now, we are moving to the Buick. The cells originate as induced pluripotent stem cells (iPSCs) a relative of embryonic stem cells that is based on reprogramming adult cells.

Two Madison-based businesses, and sources at the University of WisconsinMadison, also helped to fund the research. Fujifilm Cellular Dynamics Inc., one of the largest commercial sources of stem cell products, produces the committed cardiac progenitor cells that will be tested. These committed cells are ready to transform themselves into cardiomyocytes.

Fujifilm bought CDI, a company whose founders included Kamp and UWMadison stem cell pioneer James Thomson, but the operations remain in Madison. Kamp has no ownership position but is a consultant for the company.

Raval is a founder and board chair of the second commercial supporter, Cellular Logistics, Inc., which makes a freeze-dried matrix from the same proteins that naturally holds cardiomyocytes in place in the heart. The material is called extracellular matrix (ECM) because it scaffolds cells from the outside.

When the heart pumps, internal pressures often eject would-be replacement cells through lymph channels and blood vessels. Ravals group has already shown in mice that injecting extracellular matrix proteins along with new cells creates mechanical restraints that avoid the wash-out problem.

The extra-cellular matrix to be used in the NIH grant at UWMadison helped retain stem cells (yellow dots) in a pig heart. When similar cells (blue) were injected without the matrix, the cells spilled out of the heart muscle through the needle track and lymph channels.Eric Schmuck and Amish Raval, work performed at UWMadison. Eric Schmuck and Amish Raval, work performed at UWMadison

The injected scaffold may have another advantage for regenerating muscle after heart attack, Kamp notes. The ECM replenishes the scarred area to become more hospitable to the replacement cardiomyocytes. The effect may be based on chemical and mechanical signaling between the ECM and the regenerating cells.

Pigs hearts are quite close to human hearts in size and structure. The grant will cover tests on four groups of 12 pigs each following myocardial infarction:

If the combination is effective, Raval adds, We plan to proceed toward a Food and Drug Administration application for an investigational new drug, which would allow us to begin human trials.

With the passion and concern of a working cardiac surgeon, Raval says those trials would focus on patients who have not been helped by the best medical management we know today and they are not candidates for heart transplant or mechanical assist devices. The only other option is palliative or hospice care.

As Raval notes, More people are surviving heart attacks, and thats great. But many are left with a scar in the heart muscle a dead zone. That scar can enlarge, and the damage can spread. So we are seeing an increasing number of patients with heart failure. Thats why we are moving forward with this project.

This research is being funded by NIH grant 1U01HL148690-01.

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Moving beyond hype: Could one-two treatment restore damaged heart muscle? - University of Wisconsin-Madison

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Instead of testing drugs on patients directly,a cutting-edge precision medicine process is creating miniature beating hearts as primary test subjects.

Be sure to check outPart I,Part II, and Part IIIof our interview series on precision medicine!

Imagine a doctor recommends a strong medicine to a patient, a medicine that often causes cardiac problems in patients.

However, instead of testing the drug on the patient, the doctor gets a lab-grown, mini-sized replica of the patients heart.

The drugs are administered on the mini heart until the right drug is found. Only then is it administered to the patient.

Imagine a situation where you take cells from a particular patient, make these little mini hearts for that patient, and test potential therapies in the in vitro system before you subject the patient to those therapies

Before you start thinking Chromosome 6, (a reference to a Robin Cook book) see what Kevin Costa, Co-Founder and Chief Scientific Officer at Novoheart told The Sociable.

Their MyHeart platforms miniature hearts made from human tissue could be bringing in a revolution in precision medicine as well as drug discovery.

Precision medicine isnt just human-based, its individual based, and you can get increasingly precise, he says.

With a mini heart pumping away like any regular one, one can start to specialize a little bit more. For example, finding out the specifics of a disease in different ethnic backgrounds, like the Jewish population, African-American, Caucasian, or Asian.

So you can go from just having a human heart to an Asian, Caucasian, African, or whatever you want. You can also look at differences between male and female. So thats starting to get a little more precise, he explains.

So we can really get to the level of precision of an individual.

You can even make these tissues from an individual patient. Literally, weve got hearts in our laboratory that were made from a particular patients skin cells, he gushes.

Novoheart focuses on stem cell and tissue engineering for next-generation drug development and discovery. They are primarily a service company providing screening services based on their human tissue engineering technology, which they call the MyHeart platform.

Read more: Deep tech, big data, and their impact on precision medicine

The MyHeart platform consists of several different cardiac assays of human cardiomyocytes, human heart cells that are derived from human induced pluripotent stem (IPS) cells, which means human stem cells that can be differentiated into any cell type of the body.

The IPS cells are then mixed with the cardiomyocytes to make a 3D Structure and then cast into different types of tissues or layers of cells that Novoheart uses to measure electrophysiology or strips of tissue to measure contractility.

Rather than subjecting the patient to testing various cocktails of drugs, if we could get some information early on about whether a particular patient is more susceptible to a therapy, we can treat at a very granular, precise level for each patient

So its kind of like the electrical and mechanical side of how the heart works. We make these little mini hearts that pump like human hearts and give us measurements that clinicians are interested in, for example, cardiac output stroke volume he says.

Everything that Novoheart does is based on human cells. Tissue engineering has evolved to use human cells instead of rodent, and this was the basis for the original idea for Novoheart.

The company combined Costas expertise in tissue engineering in cardiac mechanics, Co-founder and CEO Ronald Lis expertise in human stem cells and cardiac electrophysiology, and Co-founder and Scientific Advisory Board member, Michelle Khines expertise in microfluidic platforms.

Drug discovery currently involves a process that starts with investigating a few thousand compounds in the laboratory, from which a couple of hundred that look promising can be impressed in an animal model.

Then you have to sort of take a leap of faith in moving from testing on animals to human patients. Thats the next step in the clinical trial process, Costa explains.

Costa says for every drug that enters a clinical trial process, 90% of them fail. Maybe, one out of ten that goes back out of several hundred is actually a go, after which clinicians consider candidates and try to get FDA approval.

Its a very inefficient and time-consuming process involving a couple of billion US dollars. Typically, to go from initial concept to approval, it can take over a decade, he says.

The Novoheart team thinks that part of the inefficiency lies in that leap of faith in going from animals to patients.

The way to help improve that process would be if we could get information in a human based heart system before actually testing on patients.

Novoheart has found a less risky way in terms of safety concerns for trying things on patients for the first time.

Also, if a compound doesnt work, you can reiterate in the laboratory and improve its safety and efficacy before moving on to clinical trials. This could ensure an increase in the success rate of clinical trials from 10% to who knows 50% or more.

According to Costa, one of the top reasons that drugs fail in the regulatory approval process is because of cardiac side effects, which is a major roadblock. That is a part of the reason Novoheart focuses on cardiac miniatures.

We focus on cardiac because thats our expertise. But the drugs that we are testing can be for any body part or disease because they all have to go through at least a cardiac safety assessment, he says.

They make two classes of heart tissue, a healthy heart tissue as well as diseased ones.

These organoids are designed thinking ahead towards that day when we will be able to have a little heart organoid, a liver organoid and a little brain organoid, all communicating with one another, kind of like a little humanoid

If you want to find a drug thats going to cure diabetes, you want to ensure it isnt going to give you heart disease in the process. So you can try it on the healthy heart tissue and see if its safe. If it causes arrhythmias or hypertrophy, it would be a problem for the patient, he says.

The other kind of tissue they make is diseased tissue.

If youre trying to develop a drug to cure heart disease, you need to have a model of that disease. So Novoheart is actively involved in that as well, he says.

Will they branch out then to other organoids? How about the liver?

Read more: Machine learning will be able to predict diseases years before onset of symptoms

Costa says Novoheart is thinking about combining different types of organoids together with the technology theyve developed. Costa paints a little picture of the future,

These organoids are designed thinking ahead towards that day when we will be able to have a little heart organoid, a liver organoid and a little brain organoid, all communicating with one another, kind of like a little humanoid.

Currently, its not particularly cost-effective to be able to do this for every single patient. However, Costa says, as the process becomes more streamlined and economical, the future is hopeful.

Imagine a situation where you take cells from a particular patient, make these little mini hearts for that patient, and test potential therapies in the in vitro system before you subject the patient to those therapies, he says.

Precision medicine isnt just human-based, its individual based, and you can get increasingly precise

This will have a major impact on medicine because, often, a cardiologist has to consider multiple therapies for a patient. In the current way of doing things, they try out and see what works on the patient. If not, they go to a second trial, second drug, and see what works best. If this testing process could instead be done on little organoids, it would be helpful.

Not just cardiac drugs, many chemotherapies have cardiac side effects. So rather than subjecting the patient to testing various cocktails of drugs, if we could get some information early on about whether a particular patient is more susceptible to a therapy, we can treat at a very granular, precise level for each patient, he says.

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How growing mini human hearts is advancing precision medicine, drug discovery - The Sociable

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Improving on Current Immunotherapies

Researchers from the Wellcome Sanger Institute, GlaxoSmithKline and Biogen, working under the Open Targets initiative, have demonstrated that thousands of DNA differences that are associated with immune diseases are also connected to the specific switching on of a subtype of immune cells. Previous research has shown that there are thousands of genetic variants common in individuals with immune diseases. The new study was published in the journal Nature Genetics.

Our study is the first in-depth analysis of immune cells and cytokine signals in the context of genetic differences linked to immune diseases, said Blagoje Soskic, lead author of the paper from the Wellcome Sanger Institute and Open Targets. We found links between the disease variants and early activation of memory T-cells, suggesting that problems with regulating this early T-cell activation could lead to immune diseases.

The research teams evaluated parts of the genome active in three types of immune cells accumulated from healthy volunteers, then compared their positions against all the genetic variants associated to different immune diseases. They also added different cytokines to their pool, which then consisted of a total of 55 different cell states that mimic immune disease inflammation. One particular cell type and cell state, early activation of memory T-cells, had the most active DNA as the same regions as the genetic variants associated with immune diseases. Cytokines, however, did not have as significant a role as suspected.

There are thousands of different cell types and states in the body, and finding the cause of autoimmune diseases is like finding a needle in a haystack, said Gosia Trynka, senior author from Wellcome Sanger Institute and Open Targets. We have identified early activation of memory T-cells as being particularly relevant to immune diseases, and will now be able to dive deeper into studying how this is regulated, to discover genes and pathways that could be used as drug targets.

Treating Heart Attacks with an Injectable Hydrogen

Researchers at the University of California San Diego, showed that use of an injectable hydrogel was able to repair damage and restore heart function in patients after a heart attack. This was a Phase I clinical trial sponsored by Ventrix, a UCSD spin-off. The gel, named VentriGel, is manufactured from cardiac connective tissue from pigs. The researchers take the tissue, strip out heart muscle cells, then freeze-dry and grind it into powder, then liquefy into a fluid that allows it to be injected into the heart muscle without surgery. At room temperate the liquid become a semi-solid, porous gel.

The Chromosome Connections Between Humans and Archaebacteria

Archaebacteria are some of the oldest-living organisms on the planet, one of the three biological domains: bacteria, eukaryotes, and archaea. Researchers at the University of Indiana found that the way DNA is organized in archaeal chromosomes has more similarities to human DNA than it does to bacteria. They believe it may help scientists study human DNA and diseases more effectively because the archaea are similar but less complex than human DNA.

How the 2 Strands of DNA are Held Together

DNA is made up of two strands of sugar and phosphate molecules, twisted into a helix. It has been generally accepted that the two strands were held together by hydrogen bondswhich now appears to be incorrect. Researchers at Chalmers University of Technology, Sweden, showed that that molecules have a hydrophobic interior and exist in an environment mostly of watermeaning that the DNA molecules nitrogen basis are hydrophobic, pushing the surrounding water away. The hydrogen bonds appear be more involved in sorting the base pairs rather than connecting the two strands together.

Unexpected Amyotrophic Lateral Sclerosis Findings

Accumulation of a protein, TDP-43, in the brain has been linked to amyotrophic lateral sclerosis (ALS). Researchers used a technique called deep mutagenesis to study all possible mutations in the TDP-43 protein with unexpected results. They developed more than 50,000 mutations of TDP-43 and tracked their toxicity to yeast cells. However, instead of finding the mutant forms to be more toxic, they were less toxic, forming unusual liquid species in the cells. Although unclear, the researchers believe its possible that aggregation of TDP-43 is actually protective, rather than damaging.

Antimicrobial Resistance is Growing Dramatically Around the World

Researchers developed a map of the world showing incidences of antimicrobial activity. The overall picture is of a dramatic increase in antimicrobial resistance, with the highest rates in animals in northeast China, northeast India, southern Brazil, Iran and Turkey. Some of this is related to improved economies, leading to increased meat consumption, linked to dramatic increases in the use of antibiotics in farm animals, but in countries with lower rates of monitoring for antibiotic resistance.

Possible New Weapon for Use in Immunotherapy for Cancer: iNKT Cells

Invariant natural killer T-cells (iNKT) are not as common as other types of immune cells, but they are generally viewed as more powerful. Researchers at UCLA, working in mice, were able to harness iNKT cells to attack cancer cells. They genetically engineered hematopoietic stem cells to develop into iNKT cells, which they then tested on mice with both human bone marrow and human cancers and multiple myeloma and melanoma models. The stem cells differentiated normally into iNKT cells and continued to produce them for the rest of the animals lives. The stem cell-derived iNKT cells also effectively suppressed tumor growth.

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Research Roundup: Improving Current Immunotherapies and More - BioSpace

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Researchers Build Microscopic Biohybrid Robots Propelled by Muscles and NervesArnold Lander posted on September 27, 2019 |

An artistic rendering of a new generation of biobotssoft robotic devices powered by skeletal muscle tissue that is stimulated by onboard motor neurons. (Image courtesy of Michael Vincent.)

Researchers at the University of Illinois have developed a biohybrid robot powered by neuromuscular tissue that responds to light. Biohybrid robots are the result of integrating synthetic material and living tissue such as muscle, nerves or bone to produce a device that is capable of independent motion. The addition of neuronal action to control muscle tissue represents a significant step forward in the quest for autonomous biobots.

In 2014 researchers developed the first self-propelled biobots powered by cardiac muscle tissue taken from rats. These early designs, modeled after sperm cells, had a single tail and could swim but could not sense their environment or make decisions.

In this new study, computational models were used to optimize the skeleton design. The previous single-tailed structure was replaced with a new two-tailed model, and the length of the tails was also adjusted. These design improvements resulted in an order of magnitude increase in swimming speed from the previous single-tailed version.

The robot was completed by applying an optogenetic cell culture derived from mouse stem cells adjacent to the muscle tissue. In this process, the neurons advanced toward the muscle and formed neural muscular junctions, with the robot assembling entirely on its own.

The biobot team: (from left) Professor Tahir Saif, graduate student Omar Aydin, graduate student Xiastian Zhang, Professor Mattia Gazzola, graduate student Gelson J. Pagan-Diaz, and Dean of Granger College of Engineering, Rashid Bashir.

The success of this study helps set the stage for the future development of engineered, multicellular living systems with the ability to respond intelligently to environmental cues. These living machines could potentially find applications in the fields of bioengineering, medicine and material science.

The paper Neuromuscular actuation of biohybrid motile bots is availablehere.

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Researchers Build Microscopic Biohybrid Robots Propelled by Muscles and Nerves - ENGINEERING.com

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Abstract

The control of stem and progenitor cell fate is emerging as a compelling urgency for regenerative medicine. Here, we propose a innovative strategy to gain optical control of endothelial colony-forming cell fate, which represents the only known truly endothelial precursor showing robust in vitro proliferation and overwhelming vessel formation in vivo. We combine conjugated polymers, used as photo-actuators, with the advantages offered by optical stimulation over current electromechanical and chemical stimulation approaches. Light modulation provides unprecedented spatial and temporal resolution, permitting at the same time lower invasiveness and higher selectivity. We demonstrate that polymer-mediated optical excitation induces a robust enhancement of proliferation and lumen formation in vitro. We identify the underlying biophysical pathway as due to light-induced activation of TRPV1 channel. Altogether, our results represent an effective way to induce angiogenesis in vitro, which represents the proof of principle to improve the outcome of autologous cell-based therapy in vivo.

In recent years, organic semiconductors have emerged as highly promising materials in biotechnology, thanks to several key-enabling features. Differently from silicon-based electronics, they support both electronic and ionic charge transport (1); they can be easily functionalized with specific excitation and sensing capabilities (24); and they are solution processable, soft, and conformable (5). They are highly biocompatible, being suitable for in vivo implantation and long-term operation, as recently reported for many different applications, including electrocorticography, precise delivery of neurotransmitters, electrocardiography, deep brain stimulation, and spinal cord injury (69). An important, distinctive feature of organic semiconductors is their sensitivity to the visible and near-infrared light. Recently, our and other groups have exploited it for optical modulation of cell electrophysiological activity, by using conjugated polymers and organic molecules as exogenous light-sensitive actuators (1012). Interesting applications have been reported in the field of artificial visual prosthesis (5), photothermal excitation or inhibition of cellular activity (13, 14), and modulation of animal behavior (15).

In this framework, the opportunity to use polymer-based phototransduction mechanisms to regulate the very early stages of living cell development has been very scarcely considered (16, 17). The possibility to selectively and precisely regulate a number of cell processes, such as adhesion, differentiation, proliferation, and migration, would be key to regenerative medicine and drug screening. The presently dominant approaches to reliably regulate stem and progenitor cell fate for regenerative purposes mainly rely on the use of chemical cues. However, irreversibility and lack of spatial selectivity represent important limitations of these methods. Whenever targeting in vivo applications, one must face the major, unsolved problem of diffusion of neurotrophic molecules by the conventional intravenous or oral routes. In addition, the therapeutic outcome of autologous cell-based therapy is often impaired by low engraftment, survival, and poor integration of stem cells within the environment of the targeted tissue. Other stimuli, mainly consisting of mechanical and electrical cues, were recently reported to have some notable effects, and recent advances in nanotechnology and material science enabled versatile, robust, and larger-scale modulation of the cell fate. In particular, carbon-based materials and conjugated polymers led to interesting results (18). However, their distinctive visible light absorption was never exploited in optically driven techniques.

Use of light actuation has been proposed either by viral transfer of light-sensitive proteins, by optogenetics tools, or by absorption of endogenously expressed light-sensitive moieties, based on low lightlevel therapies (1921). In the first case, interesting results were obtained (22); however, this approach bears all the drawbacks related to the need for viral gene transfer. Photobiomodulation led to interesting outputs as well, but overall efficiency is hampered by the limited absorption of light-responsive molecules endogenously expressed in living cells.

In this work, we propose to couple the use of conjugated polymers with visible light excitation to gain optical control of cell fate. We focus our attention on endothelial progenitor cells (EPCs) and, in particular, on endothelial colony-forming cells (ECFCs), which are currently considered the bona fide best surrogate of EPCs (23). ECFCs are mobilized from the bone marrow and vascular stem cell niche to reconstruct the vascular network destroyed by an ischemic insult and to restore local blood perfusion (24). ECFCs may be easily harvested from peripheral blood, display robust clonogenic potential, exhibit tube-forming capacity in vitro, and generate vessel-like structures in vivo (24, 25), thereby representing a promising candidate for autologous cell-based therapy of ischemic disorders (24). Manipulating the signaling pathways that drive ECFC proliferation, migration, differentiation, and tubulogenesis could represent a reliable strategy to improve the regenerative outcome of therapeutic angiogenesis in the harsh microenvironment of an ischemic tissue, such as the infarcted heart (24, 25). Intracellular Ca2+ signals play a crucial role in stimulating ECFC proliferation and tubulogenesis by promoting the nuclear translocation of the Ca2+-sensitive nuclear transcription factor B (NF-B) (2628). It has, therefore, been suggested that intracellular Ca2+ signaling could be targeted to boost the regenerative potential of autologous ECFCs for regenerative purposes (29). For the above-mentioned reasons, ECFCs represent a valuable test bed model for assessing the possibility to exploit the visible light sensitivity of conjugated polymers to gain touchless, optical modulation of cell proliferation and function.

In this framework, we demonstrate that polymer-mediated optical excitation during the first steps of ECFC growth leads to a robust enhancement of both proliferation and tubulogenesis through the optical modulation of the Ca2+-permeable transient receptor potential vanilloid 1 (TRPV1) channel and NF-Bmediated gene expression. Our results represent, to the best of our knowledge, the first report on the use of polymer photoexcitation for the in vitro modulation of ECFC fate and function, thereby representing the proof of principle to obtain direct control of progenitor cell fate.

Figure 1A shows a sketch of the bio/polymer interface developed for obtaining optical control of ECFC proliferation and network formation, together with the polymer chemical structure and the optical absorption spectrum. The material of choice for light absorption and phototransduction is a workhorse organic semiconductor, widely used in photovoltaic and photodetection applications, namely, regioregular poly(3-hexyl-thiophene) (P3HT) (6). It is characterized by a broad optical absorption spectrum, in the blue-green visible region, peaking at 520 nm. P3HT outstanding biocompatibility properties have been reported in a number of different systems, both in vitro and in vivo, including astrocytes (30), primary neurons and brain slices (14), and invertebrate models of Hydra vulgaris (15). Chronical implantation of P3HT-based devices in the rat subretinal space did not show substantial inflammatory reactions up to 6 months in vivo (10). Here, polymer thin films (approximate thickness, 150 nm) have been deposited by spin coating on top of polished glass substrates, as detailed in Materials and Methods. Both polymer-coated and glass substrates have been thermally sterilized (120C, 2 hours), coated with fibronectin, and, lastly, used as light-sensitive and control cell culturing substrates, respectively. ECFCs have been isolated from peripheral blood samples of human volunteers and seeded on top of polymer and glass substrates.

(A) P3HT polymer optical absorption spectrum. Insets show the chemical structure of the conjugated polymer and a sketch of the polymer device used for cell optical activation. ECFCs are cultured on top of P3HT thin films, deposited on glass substrates. (B) ECFC viability at fixed time points after plating (24, 48, and 72 hours). Cell cultures were kept in dark conditions at controlled temperature (37C) and fixed CO2 levels (5%). No statistically significant difference was observed between the glass and polymer substrates at any fixed time point (unpaired Students t test). (C) Experimental setup and optical excitation protocol for evaluation of polymer-mediated cell photoexcitation effects on cell fate. Polymer and control samples are positioned within a sterilized, home-designed petri holder. Light scattering effects are completely screened. The geometry and the photoexcitation protocol have been implemented to minimize overheating effects and to keep the overall extracellular bath temperature fairly unaltered. Thirty-millisecond-long green light pulses are followed by 70 ms in dark condition.

ECFC proliferation on polymer substrates has been primarily assessed in dark conditions at three different time points, namely, 24, 48, and 72 hours after plating (Fig. 1B). Polymer-coated samples, while showing from the very beginning a slightly lower number of cells as compared with control substrates, exhibit a proliferation rate fully similar to cells plated on glass substrates (slope of the linear fitting is 0.034 0.003, R2 = 0.99 and 0.034 0.005, R2 = 0.96 for control and P3HT polymer samples, respectively).

Once assessed that the P3HT polymer surface represents a nicely biocompatible substrate for ECFC seeding and proliferation in the dark, we moved to investigate the effect of polymer photoexcitation. In more detail, to evaluate the effect of optical stimulation on cell proliferation and network formation, we continuously shined light for the whole temporal window required for cell growth, and we realized an ad hoc system suitable for operation within the cell incubator. The experimental configuration and the excitation protocol are schematically represented in Fig. 1C. Optical excitation is provided by a light-emitting diode (LED) source, with maximum emission wavelength at 525 nm, incident from the substrate side. The choice of the protocol, continuously administered to the cell cultures during early seeding and proliferation stages, has been mainly dictated by the need to avoid overheating effects, with possible negative outcomes on the overall cell culture viability. On the basis of these considerations, we opted for a protocol based on 30-ms excitation pulses, followed by a 70-ms dark condition, at a photoexcitation density of 40 mW/cm2. The whole protocol is continuously repeated for a minimum of 4 up to 36 hours, depending on the type of functional assay, at controlled temperature (37C) and CO2 levels (5%).

The temporally precise and spatially localized measurement of the temperature variation upon polymer photoexcitation at the polymer/cell interface (i.e., within the cell cleft) is not straightforward because it requires the use of localized, submicrometer probes with a fast response time. However, according to the heat diffusion equation, we expect that dissipation occurs within a few milliseconds, following exponential decrease dynamics (14). Moreover, we used the well-known method of the calibrated pipette (31) to characterize the temperature variation dynamics within the extracellular bath volume, defined by the cylinder with the base area corresponding to the light spot size and the height of about 1 m. This choice is a good approximation of the overall volume occupied by a single ECFC cell; thus, it provides a realistic estimation of the average heating experienced by the cell (fig. S1A). We observe that temperature variation closely follows short optical pulse dynamics, reaching a maximum temperature at the end of the 30-ms illumination period, quickly followed by an almost complete thermal relaxation to the basal temperature during the 70-ms-long dark period. We conclude that our polymer-based system provides a highly spatially and temporally resolved method for optical excitation, making it possible, in perspective, to selectively target single cells and even cell subcompartments. Upon prolonged illumination (hours), one should also consider possible overheating effects of the whole extracellular medium volume. The average temperature of the bath for the entire duration of the long-term experiment was measured by a thermocouple immersed in the medium. Data show that an equilibrium situation is established after 5 hours and that the absolute temperature of the bath is increased by about 1.5 (fig. S1B). The adopted prolonged excitation protocol does not negatively affect overall cell culture viability (see below).

Figure 2 reports specific effects mediated by P3HT substrates and visible light stimulation on ECFC proliferation. ECFCs were plated in the presence of EGM-2 medium to facilitate the adhesion to the substrate. After 12 hours, the medium was switched to EBM-2 supplemented with 2% fetal bovine serum, and the cells were subjected to the long-term lighting protocol for 36 hours at controlled temperature (37C) and CO2 levels (5%). Under these conditions, ECFCs seeded on P3HT and subjected to light stimulation undergo a significant increase in proliferation rate, as compared with the control condition, i.e., to cells also seeded on P3HT polymer substrates but kept in dark conditions for the whole duration of the experiment (+158% versus P3HT dark; P < 0.05). No statistically significant difference in proliferation was observed among cells seeded on glass, whether they were subjected to optical excitation or not (Fig. 2A).

(A) Relative variation of the proliferation rate of ECFCs subjected to long-term optical excitation seeded on both bare glass and P3HT thin films, together with corresponding control samples kept in dark conditions. Cell proliferation was measured after 36 hours of culture in the presence of EBM-2 supplemented with 2% fetal calf serum. (B) Relative variation of the proliferation rate of ECFCs subjected to long-term optical excitation seeded on P3HT in the absence (CTRL) and presence of 10 M capsazepine (CPZ), 10 M ruthenium red (RR), 20 M RN-1734 (RN-1734), and 30 M BAPTA-AM (BAPTA). The results are represented as the means standard error of the mean (SEM) of three different experiments conducted on cells harvested from three different donors. The significance of differences was evaluated with one-way analysis of variance (ANOVA) coupled with Tukey (A) or Dunnetts (B) post hoc test. *P < 0.05.

Recent evidence demonstrated an interesting correlation between processes key to ECFC vascular regeneration, including proliferation and network formation, and activation of TRPV1 channels, which are expected to be endogenously expressed in ECFCs (32). In addition, we recently reported that polymer photoexcitation leads to selective TRPV1 activation in transfected human embryonic kidney (HEK) cell models (33). Therefore, we were prompted to evaluate whether the increase in cell proliferation is distinctively determined by a polymer-mediated photoactivation of the TRPV1 channel. To this goal, we preliminarily checked the actual expression of the TRPV1 channel in the ECFC models by carrying out electrophysiology experiments in patch-clamp configuration. Methods and results are extensively discussed in the Supplementary Materials (fig. S2 and related description). Briefly, the expression of the TRPV1 channel was confirmed, as well as the capability to selectively excite its activity through localized polymer excitation at high optical power density. To establish whether the TRPV1 channel also has a role in the observed increase in cell proliferation upon polymer excitation, we performed the experiments under light illumination upon administration of a highly specific TRPV1 antagonist [capsazepine (CPZ), 10 M], an aspecific TRPV channel inhibitor [ruthenium red (RR), 10 M], and a selective antagonist of a different temperature-sensitive channel, TRPV4, which is also endogenously expressed in ECFCs (RN-1734, 20 M) (34) (Fig. 2B). TRPV1 inactivation by CPZ and RR results in a relative, strong reduction in cell proliferation by 51 and 30%, respectively, as compared with untreated cells. Conversely, in the case of RN-1734 treatment, the proliferation increase due to polymer photoexcitation is completely unaltered.

As mentioned earlier, intracellular Ca2+ signaling has been reported to drive ECFC proliferation (26, 28). To further investigate whether TRPV1-mediated extracellular Ca2+ entry mediates the proangiogenic response to light illumination, we pretreated ECFCs with [1,2-Bis(2-aminophenoxy)ethane-N,N,N,N-tetraacetic acid tetrakis(acetoxymethyl ester) BAPTA-AM] (30 M), a membrane-permeable buffer of intracellular Ca2+ levels (26, 28). BAPTA-AM is widely used to prevent the increase in intracellular Ca2+ concentration ([Ca2+]i) induced by extracellular stimuli and inhibits the downstream Ca2+-dependent processes. For instance, BAPTA-AM represents the most suitable tool to prevent the activation of Ca2+-sensitive decoders residing within tens of nanometers from the inner pore of plasmalemmal Ca2+ channels (35). It was recently reported that, in the absence of Ca2+-mobilizing growth factors, it does not impair the low rate of ECFC growth (27). Here, however, BAPTA-AM clearly reduced the light-driven proliferation increase, thus confirming that TRPV1 stimulates ECFCs through an increase in [Ca2+]i (Fig. 2B).

We further examined the physiological outcome of chronic light stimulation by carrying out a tube formation assay within an extracellular matrix protein-based scaffold, which is a surrogate of the basement membrane extracellular matrix. This assay recapitulates many steps of the angiogenic process, including adhesion, migration, protease activity, and tubule formation (27, 28). ECFCs were plated in the presence of EBM-2 medium supplemented with 2% fetal calf serum and subjected to the long-term lighting protocol for 8 hours at controlled levels of temperature and CO2. Control experiments carried out in dark conditions, either onto glass (see Fig. 3A for a representative optical image) or onto polymer substrates (Fig. 3C), as well as control experiments carried out upon photoexcitation of cells seeded on glass substrates (Fig. 3B), do not show remarkable differences. Conversely, ECFC cultures subjected to polymer-mediated optical excitation clearly tend to assemble into an extended bidimensional capillary-like network (Fig. 3D). Cell cultures were monitored up to 24 hours after illumination onset, but results were comparable to observations reported here, after 8 hours of illumination. This qualitative observation is fully confirmed by quantitative morphological analysis (27). As depicted in the sketch of Fig. 3E, we quantitatively evaluated the main features typical of the capillary-like network formation and, in particular, the number of master segments (Fig. 3F), master junctions (Fig. 3G), and meshes (Fig. 3H). In all cases, a notable, statistically relevant difference is observed between cells subjected to polymer-mediated optical excitation and controls. Within the same considered temporal window, the combined use of polymer substrates and visible light stimuli does not lead to sizable toxicity effects or delays in cell proliferation. Conversely, it leads to enhanced cell proliferation (Fig. 2) and allows the achievement of the formation of a more extended and mature tubular network (Fig. 3).

(A to D) Representative images of in vitro tubular networks of ECFCs subjected to long-term optical excitation seeded on both bare glass and P3HT, as well as on corresponding control samples in dark conditions. Cultures were observed up to 24 hours, but their appearance did not substantially change after pictures were taken after 8-hour culture. Scale bars, 250 m. (E) Sketch representing the main features typical of the capillary-like network that were considered for the topologic analysis. Number of master segments (F), master junctions (G), and meshes (H) analyzed in the different conditions. The results are represented as the means SEM of three different experiments conducted on cells harvested from three different donors. The significance of differences was evaluated with one-way ANOVA coupled with Tukey post hoc test. **P < 0.01 and ***P < 0.001.

As evidenced for the proliferation rate, the TRPV1 channel activation emerges to play also a fundamental role in tubulogenesis (Fig. 4). The TRPV1 pharmacological blockade with the specific inhibitor CPZ deterministically leads to a marked reduction in network formation (Fig. 4A). Upon CPZ administration, a statistically significant decrease in the relative variation of the number of master segments (Fig. 4E), master junctions (Fig. 4F), and meshes (Fig. 4G) is observed. In line with the results shown in Figs. 2 and 3, RR administration resulted in a less marked but still sizable reduction in the tubular network (Fig. 4, B and E to G), probably due to the minor specificity toward TRPV1, while the protubular effect of light remained fully unaltered in the presence of the TRPV4 inhibitor RN-1734 (Fig. 4, C and E to G). Notably, the treatment with BAPTA-AM (30 M), which affected ECFC proliferation, was able to prevent also in vitro tubulogenesis, thus corroborating the key role of intracellular Ca2+ signaling in the proangiogenic response to light illumination (Fig. 4, D and E to G). Control measurements carried out in dark conditions on polymer substrates upon the considered pharmacological treatments do not show any relevant effect (fig. S4, A to C). Overall, this evidence supports the notion that TRPV1 stimulates ECFC proliferation and network formation and demonstrates that optical excitation, properly mediated by biocompatible polymer substrates, positively affects ECFC fate by spatially and temporally selective activation of the TRPV1 channel.

(A to D) Representative optical images of in vitro tubular network of ECFCs subjected to long-term optical excitation seeded either on bare glass or on P3HT thin films and treated respectively with CPZ (A), RR (B), RN-1734 (C), and BAPTA-AM (D). Scale bars, 250 m. (E to G) Relative variation of number of master segments (E), master junctions (F), and meshes (G) of ECFCs subjected to long-term optical excitation seeded on P3HT in the absence [control (CTRL)] and presence of 10 M CPZ, 10 M RR, 20 M RN-1734 (RN-1734), and 30 M BAPTA-AM (BAPTA). The results are represented as the means SEM of three different experiments conducted on cells harvested from three different donors. The significance of differences was evaluated with one-way ANOVA coupled with Dunnetts post hoc test. *P < 0.05 and **P < 0.01.

We now turn our attention to elucidating the possible mechanisms leading to optically enhanced tubulogenesis, through TRPV1 channel activation, upon prolonged polymer excitation.

Reliable optical modulation of the cell activity mediated by polymer photoexcitation has been reported in several, previous reports, both in vitro, at the level of single cells, and in vivo, at the level of the whole animal, as evidenced by behavioral studies on both invertebrate and vertebrate models. Three different photostimulation mechanisms, active at the polymer/cell interface, have been proposed so far. These include (i) the creation of an interface capacitance, i.e., of a localized electric field, possibly affecting the cell membrane potential (11); (ii) photothermal processes, establishing a localized temperature increase upon polymer photoexcitation (13, 36); and (iii) photoelectrochemical reactions, mainly oxygen reduction processes, leading to a local variation of extracellular and/or intracellular pH (33) and sizable production of reactive oxygen species (ROS), at a nontoxic concentration, and intracellular calcium modulation (37).

In electrophysiological experiments, carried out at a photoexcitation density higher than the one used in chronic stimulation by about two orders of magnitude, we clearly observe TRPV1 excitation, corresponding however to a small variation of the cell membrane potential, in the order of a few millivolts (Supplementary Materials). Thus, upon much lower light intensity, the effects of either direct photothermal channel activation and of photocapacitive charging are expected to be negligible. To further corroborate this hypothesis, we carry out control experiments aimed at disentangling photoelectrical from photothermal transduction processes.

First, we use a different material as a cell-seeding substrate, characterized by optical absorption and heat conductivity similar to the ones typical of P3HT (13) but fully electrically inert (i.e., unable to sustain electronic charge generation upon photoexcitation). The material of choice is a photoresist (MicroPosit S1813). S1813 thin films are realized by spin coating, and deposition parameters are optimized to obtain optical absorbance values similar to the semiconducting polymer samples at the considered excitation wavelength. The capability of photoresist substrates to sustain ECFC proliferation was successfully assessed in a control measurement, obtaining fully comparable results with respect to the P3HT substrates (Fig. 5A). The functional effect eventually driven by photoresist optical excitation on tubulogenesis was then investigated by using the same experimental conditions and analysis technique previously adopted for polymer and glass substrates (Fig. 5B). Data show that long-term photoresist excitation does not lead to sizable enhancement of the cellular network formation, thus pointing out that a purely photothermal effect does not play a major role in boosting the tubulogenesis process at variance with semiconducting polymer substrates. In a complementary experiment, we directly assessed the occurrence of photoelectrochemical reactions at the polymer/extracellular bath interface by measuring ROS production. We previously demonstrated that P3HT polymer thin films exposed to saline electrolytes sustain efficient light-triggered charge generation and charge transfer processes, giving rise to photoelectrochemical reactions (38, 39). Moreover, we also reported that P3HT nanoparticles are efficiently internalized within the cytosol of secondary line cell models (HEK-293) and that their photoexcitation leads to the production of ROS and subsequent intracellular calcium modulation (15, 37). However, the actual capability to sustain photoelectrochemical reactions in the specific experimental conditions used in this work (polymer film deposition conditions, sterilization process, prolonged exposure to specific cellular growth medium in an incubating environment, prolonged exposure to a light excitation protocol, light wavelength, pulses duty cycle, and power density) was never assessed. In particular, direct measurement of intracellular ROS was never carried out in the presence of polymer thin films. To this goal, we realized ECFC cultures on top of polymer and glass control substrates, and we exposed them to the same optical stimulation protocol previously used in the tubulogenesis assay. ROS production was then evaluated by means of a fluorescence experiment based on the use of the well-known ROS probe 2,7-dichlorodihydrofluorescein diacetate (H2DCF-DA) (Fig. 5C). Results show that light induces an increase in ROS production both on glass and polymer substrates. Relative percentage variation amounts to +34 and +200%, respectively, thus pointing out that polymer surface photocatalytic activity plays a major role in the phototransduction phenomenon.

(A) An electrically insulating, thermally conducting material (photoresist) is successfully used as an ECFC seeding substrate. (B) Photoresist long-term photoexcitation does not lead to sizable enhancement in tubulogenesis parameters. (C) Evaluation of intracellular ROS production following long-term photoexcitation protocol of ECFC cultures on polymer and glass substrates (glass dark, n = 629; glass light, n = 656; P3HT dark, n = 686; and P3HT light, n = 583). For each panel, the results are represented as the means SEM of three different experiments conducted on cells harvested from three different donors. The significance of differences was evaluated with unpaired Students t test (A and B) or one-way ANOVA coupled with Tukey post hoc test (C). ***P < 0.001.

Altogether, data in Fig. 5 indicate that photoelectrochemical reactions induced by light at the interface between the organic semiconducting polymer and the extracellular bath play a key role in triggering the observed enhancement in cell network formation through indirect activation of the TRPV1 channel. The occurrence of faradaic phenomena at the polymer/bath interface may give rise to material degradation effects. The photostability of the polymer substrates was carefully checked by optical absorption, photoluminescence, and Raman spectra measurements. By treating the samples with the same experimental protocol used for cell tubulogenesis assays (photoexcitation density, pulses frequency, overall exposure duration, temperature, and humidity levels), no sign of irreversible polymer degradation was observed, as compared with nonilluminated samples (fig. S5).

The Ca2+-sensitive transcription factor NF-B might provide the missing link between the influx of Ca2+ through TRPV1 and the increase in proliferation and tubulogenesis observed in ECFCs upon photostimulation (26). We therefore monitored the nuclear translocation of the cytoplasmic p65 NF-B subunit via immunofluorescence staining and mRNA levels of a number of genes induced during tubulogenesis in an NF-Bdependent manner (26, 40) (Fig. 6). Our data indicate that ECFCs seeded on polymer and subjected to light stimulation have a significantly enhanced p65 NF-B nuclear translocation compared with the control conditions consisting of cells also seeded on P3HT but kept in dark conditions (+35% versus P3HT dark; P < 0.05; Fig. 6, A and B), and seeded on bare glass (+28% versus glass dark; P < 0.05; Fig. 6B). No differences were observed between samples seeded on glass, whether they were subjected to optical excitation or not (fig. S6).

ECFCs seeded on P3HT samples and glass controls are subjected to long-term photostimulation protocol. Corresponding control samples are kept in dark conditions. After photostimulation, p65 NF-B nuclear translocation (A and B) and mRNA levels of tubulogenic/angiogenic genes that have been shown to be activated downstream of NF-B (C) are evaluated. (A) Representative images of immunofluorescence staining showing p65 NF-B (green) nuclear translocation. Cell nuclei are detected by 4,6-diamidino-2-phenylindole (DAPI) (blue). Scale bars, 50 m. (B) Quantitative evaluation of p65 NF-B nuclear translocation, as evidenced by colocalization experiments. Results are expressed as means SEM of the relative percentage of p65 nucleipositively stained cells to the total number of cells (glass dark, n = 151; glass light, n = 125; P3HT dark, n = 147; and P3HT light, n = 159). Ten fields per condition are analyzed. Data are obtained from two different experiments conducted on cells harvested from two different donors. (C) mRNA levels of intercellular adhesion molecule 1 (ICAM1), selectin E (SELE), and matrix metalloproteinases (MMP1, MMP2, and MMP9) are quantified by real-time polymerase chain reaction (PCR). Data are expressed as means SEM of percentage variation with respect to cells grown in the dark (n = 6). The significance of differences was evaluated with unpaired Students t test (C) or one-way ANOVA coupled with Tukey post hoc test (B). *P < 0.05 and **P < 0.01.

In addition, we have checked the expression of nine genes whose expression is known to be induced in endothelial cells during tubulogenesis/angiogenesis in an NF-Bdependent manner. We considered intercellular adhesion molecule 1 (ICAM1); vascular adhesion molecule 1 (VCAM1); selectin E (SELE), matrix metalloproteinases (MMPs) 1, 2, and 9; vascular endothelial growth factor A (VEGFA); cyclooxygenase 2 (COX2, PTGS2); and cyclin D1 (CCND1) (40). Of these, five are significantly up-regulated by light exposure in cells grown on P3HT substrates, namely, ICAM1 (+90% versus P3HT dark; P < 0.05), SELE (+1119%; P < 0.01), MMP1 (+242%; P < 0.01), MMP2 (+467%; P < 0.05), and MMP9 (+458%; P < 0.05) (Fig. 6C). Conversely, VCAM1, VEGFA, PTGS2, and CCND1 do not show relevant variation upon light stimulation (fig. S7A). Light excitation on cells grown on bare glass substrates does not show any significant effect as compared with control samples in dark conditions (fig. S7B).

Therapeutic angiogenesis via autologous EPC transplantation represents a promising strategy to preserve or, at least, partially restore cardiac function after myocardial infarction (24, 41). Nevertheless, the regenerative outcome of EPC-based therapies in preclinical studies was rather disappointing and did not lead to sufficient neovascularization of the ischemic heart (41). This led to the proposal to boost their angiogenic activity by using emerging technologies, including tissue engineering of vascular niches, pharmacological preconditioning, or genetic and epigenetic reprogramming (42). ECFCs are regarded among the most suitable EPC subtypes to induce therapeutic angiogenesis and cardiac regeneration due to their high clonal proliferative potential and ability to assemble into capillary-like structures (23, 24). In addition, they can be easily isolated and expanded from the peripheral blood of patients and healthy donors. It has recently been suggested that their angiogenic activity could be boosted by targeting the intracellular Ca2+ toolkit (29). Here, we target ECFCs by adopting a fully different approach, i.e., by exploiting visible light as a modulation trigger and by the use of a thiophene-based conjugated polymer as the exogenous, light-responsive actuator. We demonstrate that photoexcitation of the organic material deterministically leads to robustly enhanced proliferation and tubulogenesis. Pharmacological assays, supported by electrophysiology experiments, allow the identification of TRPV1 selective excitation as a key player in the molecular pathway leading to macroscopic outcomes, as observed by quantitative analysis of the angiogenic response.

All data unambiguously show that polymer photoexcitation leads to selective activation of the TRPV1 channel, which has recently been shown to be expressed and drive angiogenesis in human ECFCs (32). TRPV1 is a polymodal Ca2+-permeable channel that integrates multiple chemical and physical cues to sense major changes in the local microenvironment of most mammalian cells (43). TRPV1 is activated by either noxious heat (>42C) and acidic solutions (pH < 6.5), whereas mild acidification (pH 6.3) of the extracellular milieus sensitizes TRPV1 to heat stimulation and results in channel activation at temperature thresholds (30 to 32C) well below the normal one (43). ROS production is also expected to further contribute to TRPV1 activation, as previously reported in mouse coronary endothelial cells (44), in which hydrogen peroxide elicits a depolarizing inward current at negative holding potentials. Likewise, ROS may stimulate TRPV1 to depolarize the membrane potential, thereby triggering trains of action potentials in airway C fibers (45, 46).

On the basis of measurements carried out in cells seeded on the photoresist substrate, as well as on direct evaluation of a limited, local temperature increase upon light stimuli during the long-term photoexcitation protocol, we infer that the excitation of the TRPV1 channel through direct photothermal transduction is not the predominant process leading to enhanced tubulogenesis.

We have previously demonstrated that polymer photoexcitation leads to generation of faradaic current, to electron transfer reactions at the polymer/electrolyte interface, and to sizable intracellular enhancement of ROS (37, 38). Briefly, optical excitation of P3HT polymer thin films leads to photoexcited species (Eq. 1), namely, singlets and charge states, which react with the oxygen dissolved in the cell medium, thus reducing oxygen (Eq. 2)P3HT+hP3HT*(1)P3HT*+O2P3HT++O2(2)

The superoxide further evolves, leading to the generation of different ROS and, lastly, ending up with hydrogen peroxide production. It has been reported that extracellular H2O2 can cross the plasma membrane through aquaporin AQP3, thereby triggering intracellular ROS signaling (47, 48). In line with our previous results, we have demonstrated here that intracellular ROS enhancement does occur in ECFCs upon photoexcitation of polymer thin films, thus contributing to TRPV1 activation.

Altogether, the evidence supports the hypothesis of a transduction mechanism mainly governed by photoelectrochemical reactions. Moreover, these same observations could explain why TRPV4, which is also expressed in ECFCs (34), is not sensitive to optical modulation. Although TRPV4 is activated by moderate heat (24 to 38C), it is supposed to be inhibited by local pH variation, although this is still a matter of debate (49, 50).

On the one hand, the role attributed in the phototransduction mechanism to the capability of the polymer to generate and transport electronic charges, as well as to its photocatalytic activity in an aqueous environment, clearly implies the need for a biocompatible, visible lightresponsive, semiconducting material. This excludes any possible implementation of the reported technique by using a thermally conducting, electrically insulating plastic substrate. Suitable cell-seeding materials have to be selected and developed within the wide arena of organic semiconducting polymers. On the other hand, the key role played by ROS raises additional issues about material photostability, cell viability, and overall safety and reliability of the technique. We extensively verified that the main polymer optoelectronic properties are not substantially altered by the exposure to light and to incubating conditions. From the biological point of view, it is very well known that high ROS levels can induce highly toxic effects and, finally, lead to cell death. We notice, however, that the established photoactivation protocol (illuminator geometry and air flow, light photoexcitation density, duty cycle, and repetition rate) has been implemented to avoid any detrimental effect. Accordingly, no toxicity effects were detected for the overall duration of the experiments, as proven by the robust increase in ECFC proliferation and tubulogenesis exposed to light. This observation is consistent with the emerging notion that appropriate ROS levels can exert a signaling role and control angiogenesis in endothelial cells (51).

The biophysical mechanisms whereby the photoactivation of TRPV1 stimulates in vitro angiogenesis in ECFCs deserve a more detailed discussion as well. Earlier work showed that TRPV1 stimulates proliferation and tube formation in vascular endothelial cells by mediating extracellular Ca2+ entry. The following increase in intracellular Ca2+ concentration ([Ca2+]i) leads to the recruitment of several downstream Ca2+-dependent decoders, such as endothelial nitric oxide synthase and Ca2+/calmodulin-dependent protein kinase II (CaMKII) (52). Recently, TRPV1 was found to induce also proliferation and tube formation in ECFCs by mediating the uptake of the endocannabinoid anandamide (32). This study, however, did not investigate whether TRPV1 activation was per se able to stimulate ECFCs by engaging Ca2+-dependent pathways. Intracellular Ca2+ signaling is a crucial determinant of ECFC fate and behavior (2628). Accordingly, light-induced ECFC proliferation and tube formation were markedly reduced by the pharmacological blockade of TRPV1-mediated Ca2+ entry with CPZ and RR and by preventing the subsequent increase in [Ca2+]i with BAPTA-AM. This finding endorses the view that optical excitation stimulates ECFCs through TRPV1-mediated extracellular Ca2+ entry, and we suggest here that this occurs via downstream activation of transcriptional factor NF-B. NF-B has previously been shown to stimulate cell proliferation and tubulogenesis in endothelial cells (53, 54) and in hepatocytes (55). Our group has shown that NF-B triggers the transcriptional program underlying the angiogenic response to extracellular Ca2+ entry in ECFCs (26). Moreover, NF-B activation in response to extracellular stimulation and Ca2+ entry through TRPV1 has also been demonstrated (56, 57). Under resting conditions, NF-B is retained in the cytoplasm by the complex with the inhibitory protein IB. An increase in [Ca2+]i results in IB degradation by ubiquitination, which is triggered upon the Ca2+-dependent phosphorylation of IB. As a consequence, the p65 NF-B subunit is released from IB inhibition and translocates into the nucleus (58) where it induces the expression of multiple proangiogenic genes (40). Consistently, we found that optical excitation significantly boosted the nuclear translocation of p65 in ECFCs cultured on the conjugated polymer compared with those not exposed to light. Robust up-regulation of several angiogenic genes, such as ICAM, SELE, MMP1, MMP2, and MMP9, which are under NF-Bdependent transcriptional control, was also consequently observed. Intriguingly, NF-B also mediates VEGFA-induced gene expression and angiogenesis in vascular endothelial cells (59, 60) through an increase in [Ca2+]i (61). These observations strongly hint at NF-B as the Ca2+-sensitive decoder that translates optical excitation into an angiogenic response in human ECFCs interfaced with the light-sensitive conjugated polymer.

Overall, our findings represent the proof of principle that optical modulation may be successfully exploited to directly control the fate of a progenitor cell population, i.e., ECFCs, which has been shown to support revascularization of ischemic tissues. The in vitro activation of ECFC angiogenic activity is made possible by the use of a biocompatible, light-sensitive polymer as the phototransduction element.

The combined use of optical excitation and organic polymer technology can open interesting perspectives for several different reasons. First, the use of light modulation allows unprecedented spatial and temporal resolution to be achieved in a fully reversible way. Light temporal and spatial patterns can be specifically designed and adapted to different in vitro cell models, allowing ideally endless combinations of possibilities, to finely tune overall output in cell proliferation and network formation. The demonstrated technology is minimally invasive, allows for massive parallelization of experiments, and can be virtually implemented in any cell therapy model in a straightforward way. In addition, the use of different polymers, with lower energy gap and in the form of nanobeads, may pave the way to the optical enhancement of therapeutic angiogenesis in vivo. Further work is needed to understand whether the pattern and/or intensity of the illumination protocol may be adjusted to further boost the angiogenic response. For instance, the optical excitation protocol consisted of 30-ms-long light pulses that were delivered at 1 Hz for 4 (tubulogenesis) up to 36 (proliferation) hours. This is likely to result in oscillations in [Ca2+]i, which are known to deliver the most instructive signal for ECFCs to undergo angiogenesis by inducing the nuclear translocation of the p65 NF-B subunit (26). As the frequency of intracellular Ca2+ oscillations can be artificially manipulated to regulate NF-Bdependent gene expression in virtually any cell type (62), we envisage an additional layer of specificity and control that could be exploited to further improve the angiogenic response to optical excitation. Future work will also be devoted to assess the outcome of optical modulation on patient-derived ECFCs. One of the main hurdles associated to autologous cell-based therapy is the impairment of the angiogenic activity of EPCs, including ECFCs harvested from cardiovascular patients (29). The therapeutic translation of our findings will require the demonstration that light-induced TRPV1 activation boosts angiogenesis also in ECFCs derived from individuals affected by severe cardiovascular disorders, such as hypertension, atherosclerosis, and heart failure. In this view, the combination of organic semiconductors and genetic manipulation to increase endogenous TRPV1 expression could be sufficient to restore the reparative phenotype of autologous ECFCs from cardiovascular patients.

Regioregular P3HT (99.995% purity; Mn 54,000 to 75,000 molecular weight) was purchased from Sigma-Aldrich and used without any further purification. The samples for cell cultures were prepared by spin coating on a square 18 mm by 18 mm glass (VWR International) substrates carefully rinsed in subsequent ultrasonic baths of ultrapure water, acetone, and isopropanol. P3HT solution was prepared in chlorobenzene at a final P3HT concentration of 20 g/liter and spin coated on the cleaned substrates with a two-step recipe: (i) 3 s at 800 rpm and (ii) 60 s at 1600 rpm. Polymer film thickness is about 150 nm.

Microposit S1813 photoresist was purchased from Shipley and used without any further purification. Photoresist thin films were prepared by spin coating on cleaned substrates with a two-step recipe: (i) 3 min at 300 rpm and (ii) 30 s at 2600 rpm. Parameters were adjusted to obtain homogeneous films and similar optical absorbance to the one of the polymer thin films, at the same excitation wavelength used in the long-term stimulation protocol (see below). All films were thermally treated in an oven at 120C for 2 hours for annealing and sterilization. To promote adhesion, samples were coated with fibronectin (from bovine plasma; Sigma-Aldrich) at a concentration of 2 mg/ml in phosphate-buffered saline (PBS) for at least 30 min at 37C and then rinsed with PBS.

ECFCs were isolated from peripheral blood and expanded as shown elsewhere (26). Blood samples (40 ml) collected in EDTA-containing tubes were obtained from healthy male human volunteers aged from 28 to 38 years. The Institutional Review Board at Istituto di Ricovero e Cura a Carattere Scientifico Policlinico San Matteo Foundation in Pavia approved all protocols and specifically approved this study. Informed written consent was obtained according to the Declaration of Helsinki of 1975 as revised in 2008. We focused on the so-called ECFCs, a subgroup of EPCs that are found in the CD34+ CD45 fraction of circulating mononuclear cells (MNCs), exhibit robust proliferative potential, and form capillary-like structures in vitro (23). To isolate ECFCs, MNCs were obtained from peripheral blood by density gradient centrifugation on lymphocyte separation medium for 30 min at 400g and washed twice in EBM-2 with 2% fetal calf serum. A median of 36 106 MNCs (range, 18 to 66) was plated on fibronectin-coated culture dishes (BD Biosciences) in the presence of the endothelial cell growth medium EGM-2 MV (Lonza) containing endothelial basal medium (EBM-2), 5% fetal bovine serum (FBS), recombinant human (rh) EGF, rhVEGF, recombinant human Fibroblast Growth Factor-Basic (rhFGF-B), recombinant human Insulin-like Growth Factor-1 (rhIGF-1), ascorbic acid, and heparin and maintained at 37C in 5% CO2 and humidified atmosphere. Nonadherent cells were discarded after 2 days, and thereafter, medium was changed three times a week. The outgrowth of ECFCs from adherent MNCs was characterized by the formation of a cluster of cobblestone-shaped cells. That ECFC-derived colonies belonged to the endothelial lineage was confirmed by staining with anti-CD31, anti-CD105, anti-CD144, anti-CD146, antivon Willebrand factor, anti-CD45, and anti-CD14 monoclonal antibodies and by assessment of capillary-like network formation in the in vitro tube formation assay.

For our experiments, we have mainly used endothelial cells obtained from early-passage ECFCs (P2-4, which roughly encompasses a 15- to 18-day period) with the purpose to avoid, or maximally reduce, any potential bias due to cell differentiation. However, to make sure that the phenotype of the cells did not change throughout the experiments, in the preliminary experiments, we tested the immunophenotype of ECFCs at different passages, and we found no differences. We also tested whether functional differences occurred when early (P2) and late (P6)passage ECFCs were used by testing the in vitro capacity of capillary network formation in a Cultrex assay and found no differences between early- and late-passage ECFC-derived cells (data not shown).

Electrophysiological recordings were performed using a patch-clamp setup (Axopatch 200B; Axon Instruments) coupled to an inverted microscope (Nikon Eclipse Ti). ECFCs were measured in whole-cell configuration with freshly pulled glass pipettes (3 to 6 M), filled with the following intracellular solution: 12 mM KCl, 125 mM K-gluconate, 1 mM MgCl2, 0.1 mM CaCl2, 10 mM EGTA, 10 mM Hepes, and 10 mM ATP (adenosine 5-triphosphate)Na2. The extracellular solution contained the following: 135 mM NaCl, 5.4 mM KCl, 5 mM Hepes, 10 mM glucose, 1.8 mM CaCl2, 1 mM MgCl2. Only single cells were selected for recordings. Acquisition was performed with the pCLAMP 10 software (Axon Instruments). Membrane currents were low pass filtered at 2 kHz and digitized with a sampling rate of 10 kHz (Digidata 1440 A; Molecular Devices). Data were analyzed with Clampfit (Axon Instruments) and Origin 8.0 (OriginLab Corporation).

For optical excitation of the polymer, a homemade petri cell culture illuminator, compatible with the use within the cell incubator, was designed and implemented. Its design included a black spacer made by fused filament fabrication, both to minimize overheating effects in the extracellular bath and to avoid unwanted light scattering/diffusion effects and cross-talk between different specimens. Optical excitation was provided by a green LED system, whose duty cycle, repetition rate, and intensity were set through a custom-made control circuit, comprising a microcontroller, a digital-to-analog converter, and an analog LED driver. The driver was connected to five green LEDs (SMB1N-525V-02; Roithner LaserTechnik GmbH, Vienna, Austria), with maximum emission wavelength at 525 nm, each carrying a collimator lens reducing the emission angle to 22. This way, up to five 3.5-cm petri dishes can be simultaneously treated with a homogeneous photoexcitation density of 40 mW/cm2. The long-term optical excitation protocol adopted for cell fate modulation consists of 30-ms-long pulses, followed by 70-ms-long dark conditions, continuously repeated for a minimum of 4 up to 36 hours in the case of tubulogenesis and proliferation assays, respectively.

Growth dynamics were evaluated by plating a total of 5 103 ECFC-derived cells into 10-mm fibronectin-treated cloning cylinders (5 104/cm2) in the presence of EGM-2 MV medium to facilitate the adhesion. After 12 hours, the medium was switched to EBM-2 supplemented with 2% fetal calf serum. For the pharmacological treatment, one of compounds was added to the medium: BAPTA (30 M), CPZ (10 M), RN-1734 (20 M), or RR (10 M). Cultures were incubated at 37C (in 5% CO2 and humidified atmosphere), and cell growth was assessed after 36 hours since the beginning of the long-term illumination protocol. At this point, cells were recovered by trypsinization from all the dishes, and the cell number was assessed by counting in a hemocytometer. Preliminary experiments showed no unspecific or toxic effect for each agent when used at these concentrations. Each assay was repeated in triplicate.

ECFC-derived cells from early-passage (P2 to P4) cultures were obtained by trypsinization and resuspended in EBM-2 supplemented with 2% FBS. EPC-derived cells (10 103) per well were plated in Cultrex basement membrane extract (Trevigen Inc., Gaithersburg, MD, USA) 10-mm fibronectin-treated cloning cylinders. Plates were then incubated at 37C, 5% CO2, and capillary network formation was assessed starting from 4 to 24 hours later. At least three different sets of cultures were performed every experimental point. Quantification of tubular structures was performed after 8 hours of incubation by measuring the total length of structures per field with the aid of the ImageJ software (National Institutes of Health, USA; http://rsbweb.nih.gov/ij/). To evaluate the role of TRPV1, the same protocol was repeated in the presence of the following drugs: BAPTA (30 M), CPZ (10 M), RN-1734 (20 M), or RR (10 M).

H2DCF-DA (Sigma-Aldrich) was used for the intracellular detection of ROS. ECFCs were seeded onto polymer and control substrates and subjected to the same photoexcitation protocol used for the in vitro tube formation assay. Immediately after the end of the protocol, cell cultures were incubated with the ROS probe for 30 min. After careful washout of the excess probe from the extracellular medium, the fluorescence of the probe was recorded (excitation/emission wavelengths, 490/520 nm; integration time, 70 ms for H2DCF-DA) with an inverted microscope (Nikon Eclipse Ti) equipped with an Analog-WDM Camera (CoolSNAP MYO, Teledyne Photometrics). To minimize the effects of the spectral overlap between the polymer absorption and emission spectra, and the probe emission, samples were turned upside down by using a homemade chamber with a 500-m-thick channel filled with extracellular medium. Variation of fluorescence intensity was evaluated over regions of interest covering single-cell areas, and reported values represent the average over multiple cells. See figure captions for additional details about statistical analysis. Image processing was carried out with ImageJ and subsequently analyzed with Origin 8.0.

Two sets of P3HT thin films (n = 12) were prepared as described above. The optical absorbance, the emission, and the Raman spectrum were measured immediately after fabrication. Then, all samples were exposed to ECFC growth medium (EBM-2 supplemented with 2% FBS) and incubated at 37C, 5% CO2 for 24 hours. The first set was taken in dark conditions (n = 6), and the second one was treated with the same optical excitation protocol used in the tubulogenesis assays (n = 6). After incubation, absorption, emission, and Raman spectrum were measured again in the same conditions as before. Absorption spectra were recorded by using a spectrophotometer (PerkinElmer Lambda 1040) in transmission mode. Photoluminescence spectra were acquired by using a Jobin-Yvon spectrofluorometer; the excitation wavelength was set at the polymer absorption peak wavelength (530 nm). Resonant Raman spectra were recorded by using visible light excitation at 532 nm (HORIBA Jobin-Yvon HR800 micro-Raman spectrometer system). Laser power intensity on the sample was kept at values lower than 0.03 mW to avoid laser-induced sample degradation. Spectra were typically recorded in the region 600 to 2000 cm1 and were calibrated against the 520.5 cm1 line of an internal silicon wafer. The signal-to-noise ratio was enhanced by repeated acquisitions (100). The measurements were conducted at room temperature (RT), and the resulting spectral resolution was 0.4 cm1.

To examine NF-B p65 subunit translocation into the nucleus in the individual ECFCs, the coverslips were fixed with 4% formaldehyde in PBS (20 min at RT) and permeabilized with 0.1% Triton X-100 in PBS (7 min at RT). Primary rabbit polyclonal anti-p65 antibody (Santa Cruz Biotechnology, catalog no. Sc-372) was applied at a final dilution of 1:100 for 1 hour at 37C in a humidified chamber. After three washes with PBS, secondary chicken anti-rabbit Alexa(488)-conjugated antibody (1:200; Invitrogen, catalog no. A-21441) was applied for 1 hour at RT. After washing (three times in PBS), nuclei were counterstained with 4,6-diamidino-2-phenylindole, dihydrochloride (DAPI; 1:5000 dilution in PBS; 20 min at RT; Invitrogen, catalog no. D1306). Last, the coverslips with cells were mounted on microscope glass slides using Fluoroshield mount medium (Sigma, catalog no. F6182). Fluorescence images were taken with the same fluorescence microscope used for the electrophysiology experiments, using standard DAPI and fluorescein isothiocyanate filters set for the acquisition of DAPI and Alexa(488) fluorescence emission, respectively.

Cells were lysed in 0.5 ml of TRI Reagent (Sigma, catalog no. T9424), and total RNA was extracted according to the manufacturers protocol. One microgram of total RNA was retrotranscribed using SensiFAST cDNA Synthesis Kit (Bioline, London, UK, catalog no. BIO-65054). Real-time polymerase chain reaction (PCR) was performed using iTaq qPCR master mix according to the manufacturers instructions (Bio-Rad, Segrate, Italy, catalog no. 1725124) on a SFX96 Real-Time System (Bio-Rad). As a control, S18 ribosomal subunit was used, whose expression did not change across the conditions. For each gene, Ct was calculated by using the formula Ct = 2^(Ct(gene) Ct(S18)). The data are expressed as a percentage variation between P3HT light and glass light conditions and P3HT dark and glass dark samples, respectively. Sequences of oligonucleotide primers are listed in table S1.

The significance of differences was evaluated with unpaired Students t test or one-way analysis of variance (ANOVA) coupled with Tukey or Dunnetts post hoc test, as appropriate. Data are represented as means standard error of the mean (SEM). P < 0.05 was considered statistically significant. Statistical analysis was performed using the GraphPad Prism 7 software (GraphPad Software Inc., La Jolla, CA).

Supplementary material for this article is available at http://advances.sciencemag.org/cgi/content/full/5/9/eaav4620/DC1

Fig. S1. Local and global evaluation of the extracellular bath temperature.

Fig. S2. TRPV1 is endogenously expressed in ECFCs, and it is efficiently activated by polymer photostimulation.

Fig. S3. Current clamp measurements in HEK-293 cells.

Fig. S4. Pharmacological study on ECFCs seeded on polymer substrates in the darkEvaluation of effect on tubulogenesis.

Fig. S5. Polymer photostability.

Fig. S6. p65 NF-B nuclear translocation is unaltered in ECFCs seeded on glass subjected to light-induced photostimulation.

Fig. S7. mRNA levels of proangiogenic genes downstream of NF-B signaling.

Table S1. List of oligonucleotide primers used for real-time PCR.

This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license, which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited.

Acknowledgments: We gratefully thank I. Abdel Aziz for the characterization of the homemade petri cell culture illuminator used for long-term optical excitation and P. Falvo for the constructive criticism of the manuscript and the helpful scientific discussions. Funding: This work was jointly supported by the European Research Council (ERC) under the European Unions Horizon 2020 research and innovation program LINCE, grant agreement no. 803621 (M.R.A.), the EU Horizon 2020 FETOPEN-2018-2020 Programme LION-HEARTED, grant agreement no. 828984 (F.L., F.M., and M.R.A.), the Italian Ministry of Education, University and Research (MIUR): Dipartimenti di Eccellenza Program (20182022)Department of Biology and Biotechnology L. Spallanzani, University of Pavia (F.M.), and Fondo Ricerca Giovani from the University of Pavia (F.M.). Author contributions: F.L., F.M., and M.R.A. planned the experiments. F.L. carried out the experimental measurements (electrophysiology, short- and long-term photoexcitation, evaluation of effects on proliferation, tubulogenesis, and ROS production). V.R. provided the ECFC models, took care of the cell cultures, and contributed to the tubulogenesis and proliferation experiments. G.T. prepared the polymer samples. A.D. designed, realized, and optimized the experimental setup for the long-term photoexcitation. L.T. and D.L. carried out the immunofluorescence and real-time PCR assays. P.C. contributed to the methodological discussion about gene expression. F.L. and M.R.A. wrote the main manuscript, with help from F.M. All authors contributed to the data interpretation and approved the final manuscript. Competing interests: The authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.

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Conjugated polymers optically regulate the fate of endothelial colony-forming cells - Science Advances

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Stem cells are biological cells which have the ability to distinguish into specialized cells, which are capable of cell division through mitosis. Amniotic fluid stem cells are a collective mixture of stem cells obtained from amniotic tissues and fluid. Amniotic fluid is clear, slightly yellowish liquid which surrounds the fetus during pregnancy and is discarded as medical waste during caesarean section deliveries. Amniotic fluid is a source of valuable biological material which includes stem cells which can be potentially used in cell therapy and regenerative therapies. Amniotic fluid stem cells can be developed into a different type of tissues such as cartilage, skin, cardiac nerves, bone, and muscles. Amniotic fluid stem cells are able to find the damaged joint caused by rheumatoid arthritis and differentiate tissues which are damaged. Medical conditions where no drug is able to lessen the symptoms and begin the healing process are the major target for amniotic fluid stem cell therapy. Amniotic fluid stem cells therapy is a solution to those patients who do not want to undergo surgery. Amniotic fluid has a high concentration of stem cells, cytokines, proteins and other important components. Amniotic fluid stem cell therapy is safe and effective treatment which contain growth factor helps to stimulate tissue growth, naturally reduce inflammation. Amniotic fluid also contains hyaluronic acid which acts as a lubricant and promotes cartilage growth.

With increasing technological advancement in the healthcare, amniotic fluid stem cell therapy has more advantage over the other therapy. Amniotic fluid stem cell therapy eliminates the chances of surgery and organs are regenerated, without causing any damage. These are some of the factors driving the growth of amniotic fluid stem cell therapy market over the forecast period. Increasing prevalence of chronic diseases which can be treated with the amniotic fluid stem cell therapy propel the market growth for amniotic fluid stem cell therapy, globally. Increasing funding by the government in research and development of stem cell therapy may drive the amniotic fluid stem cell therapy market growth. But, high procedure cost, difficulties in collecting the amniotic fluid and lack of reimbursement policies hinder the growth of amniotic fluid stem cell therapy market.

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The global amniotic fluid stem cell therapy market is segmented on basis of treatment, application, end user and geography: Segmentation by Treatment Allogeneic Amniotic Fluid stem cell therapy Autologous Amniotic Fluid stem cell therapy Segmentation by Application Regenerative medicines Skin Orthopedics Oncology Fetal tissue reconstruction Kidney regeneration Regeneration of neural tissue Cardiac regeneration Lung epithelial regeneration Others Drug research and development Segmentation by End User Hospital Ambulatory Surgical Centers Specialty Clinics Academic and Research Institutes Segmentation by Geography North America Latin America Europe Asia-Pacific Excluding China China Middle East & Africa

Rapid technological advancement in healthcare, and favorable results of the amniotic fluid stem cells therapy will increase the market for amniotic fluid stem cell therapy over the forecast period. Increasing public-private investment for stem cells in managing disease and improving healthcare infrastructure are expected to propel the growth of the amniotic fluid stem cell therapy market.

However, on the basis of geography, global Amniotic Fluid Stem Cell Therapy Market is segmented into six key regions viz. North America, Latin America, Europe, Asia Pacific Excluding China, China and Middle East & Africa. North America captured the largest shares in global Amniotic Fluid Stem Cell Therapy Market and is projected to continue over the forecast period owing to technological advancement in the healthcare and growing awareness among the population towards the new research and development in the stem cell therapy. Europe is expected to account for the second largest revenue share in the amniotic fluid stem cell therapy market. The Asia Pacific is anticipated to have rapid growth in near future owing to increasing healthcare set up and improving healthcare expenditure. Latin America and the Middle East and Africa account for slow growth in the market of amniotic fluid stem cell therapy due to lack of medical facilities and technical knowledge.

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Some of the key players operating in global amniotic fluid stem cell therapy market are Stem Shot, Provia Laboratories LLC, Thermo Fisher Scientific Inc. Mesoblast Ltd., Roslin Cells, Regeneus Ltd. etc. among others.

The report covers exhaustive analysis on: Amniotic Fluid Stem Cell Therapy Market Segments Amniotic Fluid Stem Cell Therapy Market Dynamics Historical Actual Market Size, 2012 2016 Amniotic Fluid Stem Cell Therapy Market Size & Forecast 2016 to 2024 Amniotic Fluid Stem Cell Therapy Market Current Trends/Issues/Challenges Competition & Companies involved Amniotic Fluid Stem Cell Therapy Market Drivers and Restraints

Regional analysis includes North America Latin America Europe Asia Pacific Excluding China China The Middle East & Africa

Report Highlights: Shifting Industry dynamics In-depth market segmentation Historical, current and projected industry size Recent industry trends Key Competition landscape Strategies of key players and product offerings Potential and niche segments/regions exhibiting promising growth A neutral perspective towards market performance

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Amniotic Fluid Stem Cell Therapy Market to Rear Excessive Growth During 2018 2026 - Herald Space

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What if CRISPR could be used not just to edit genes, but also to alter the epigenomethe vast network of chemicals and proteins that orchestrates the actions of genes? A study from biomedical engineers at Duke University describes a new CRISPR technology that could allow scientists to do just that.

The most commonly used gene editing technology, CRISPR-Cas9, uses just one Cas protein to cut DNA. In the CRISPR field, this is known as a class 2 system. Class 1 systems, by contrast, are more complicated because they rely on multiple proteins to bind to DNA and then recruit a Cas3 protein to cut it. That network of proteins is called Cascade (CRISPR-associated complex for antiviral defense).

The Duke team used a class 1 CRISPR system to edit the epigenome in cells. In a study published in Nature Biotechnology, they reported that they were able to attach gene activators to the Cascade complex and regulate levels of gene expression in cells. They also connected a repressor to Cascade to turn genes off altogether.

Synthetic Controls: Best Practices and Regulatory Perspectives

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"We have found Cascade's structure to be remarkably modular, allowing for a variety of sites to attach activators or repressors, which are great tools for altering gene expression in human cells," said Adrian Oliver, Ph.D., a postdoctoral fellow and lead author of the study, in a statement.

RELATED: A better CRISPR? RNA 'hairpins' could improve gene editing

The potential of CRISPR-Cas3 is already generating some excitement in the biopharma world. In January, Locus Biosciences inked a deal with Johnson & Johnson to develop its platform for using CRISPR-Cas3 to solve the problem of antibiotics resistance. The deal could be worth up to $818 million for Locus, a 2019 Fierce 15 company.

Meanwhile, several academic groups are investigating various ideas for improving CRISPR, including another Duke team thats focused on improving the Cas9 enzyme. In April, it described a technique for adding a short tail to the guide RNA thats used in CRISPR systems to improve the accuracy of gene cuts.

Researchers at Columbia University are hoping to sidestep DNA cutting altogether. Theyre using a transposon, or jumping gene, in a system designed to insert DNA in exact locations in the genome without cutting.

The Duke researchers working onclass 1 CRISPR technology are planning further studies to determine whether it might help solve some of the shortcomings of CRISPR-Cas9 in addressing human disease, including the risk of dangerous immune responses. They are also investigating whether the tool could be used to perform many different genome engineering tasks simultaneously.

"We know CRISPR could have a big impact on human health," said Charles Gersbach, Ph.D., Duke professor of biomedical engineering. "But we're still at the very beginning of understanding how CRISPR is going to be used, what it can do, and what systems are available to us.

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New CRISPR approach could improve gene and cell therapies - FierceBiotech

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HIV treatment has come a long way over the years, due in large part to antiretroviral drugs that stop the HIV virus from replicating in the body. This gives the immune system a chance to repair itself and stop further damage. Thanks to these amazing advances, HIV is no longer the death sentence that it was in previous decades.

However, antiretrovirals only keep HIV at bay for as long as theyre taken. Defaulting on the drugs means that the HIV virus comes back. Even worse, it can cause patients to build up resistance to the antiretrovirals so that they do not work so effectively in the future.

In other words, theres still room for improvement when it comes to treatment. Fortunately, researchers from the University of California San Diego School of Medicine are poised to provide help, courtesy of a new genetic-sequencing approach that could possibly provide a kill switch to clear out dormant HIV reservoirs inside cells.

The most exciting part of this discovery has not been seen before, Tariq Rana, professor of pediatrics and genetics at UC San Diego School of Medicine, said in a statement. By genetically modifying a long non-coding RNA, we prevent HIV recurrence in T cells and microglia upon cessation of antiretroviral treatment, suggesting that we have a potential therapeutic target to eradicate HIV and AIDS.

The work is based on the discovery of a recently emerged gene that appears to regulate HIV replication in immune cells, including macrophages, microglia, and T cells. The team refers to this as HIV-1 Enchanced LncRNA (HEAL), and it is elevated in people with HIV. By using CRISPR-Cas9 gene editing, their work suggests that it could stop HIV from recurring in the event that antiretroviral treatment is stopped.

This has the potential for [being a] cure but, [well] have to wait for animal studies, Rana told Digital Trends. As for the next steps, Rana said that future studies will determine if turning this regulator HEAL off can remove viral reservoirs, which are the key source for viral rebound when therapies are discontinued.

A paper describing the work was recently published in the journal mBio.

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CRISPR-Cas9 gene editing could one day turn off HIV virus in the body - Digital Trends

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