Eggs from men, sperm from women: Stem cell therapy may just turn reproduction upside down!  The Economic Times

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Eggs from men, sperm from women: Stem cell therapy may just turn reproduction upside down! - The Economic Times

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1976 ROBERT ETTINGER FOUNDS THE CRYONICS INSTITUTE

Then in 1976 a separate organization was formed: the Cryonics Institute, to offer cryostasis services: careful preparation, cooling, and long term patient care in liquid nitrogen.

Our goal was maximum reliability and affordability. And we achieved it. The Cryonics Institute offers clear-cut advantages over all other providers. Such as:

Our prices are lower than any other organization in fact, the most affordable prices anywhere in the world. Our minimum whole-body suspension fee is $28,000. (For members at a distance, transportation costs and local help will be additional.) Our $28,000 fee is a one-time only payment, with no subsequent charges. Its easily funded by insurance or other means, and funds the best care available for our member patients. (For last-minute cases, where the patient was not signed up beforehand, we ordinarily charge $35,000 rather than $28,000, if arrangements can be worked out at all.)

Does that lower fee mean lower quality patient care or services? No. The major part of other organizations fees are earmarked for investment provisions totally unrelated to patient care and preparation. Methods and research differ, but overall we believe our procedures and policies give a better chance for patient survival than any other organizations and this web site will show you the detailed reasons why.

See for yourself. Read our FAQ and see The CI Advantage that compares the different cryonics organizations and why we think CI gives you and those you love the best possible chance for future survival. Remember: most CI members can afford the higher prices of other organizations for themselves and their families and often do give more, in bequests and donations. But weve chosen CI because we know its our best bet. And yours.

We have a unique, proven track record of financial security and stability. Price stability too. CI is the only organization with no debt, no stockholders, and no landlords. We own our patient care facilities outright, and all our officers and directors donate their services voluntarily. Were one of the oldest cryonics organizations in existence and the only such organization that has never raised its prices, even in high-inflation times like the late 70s and early 80s. Adjusting for inflation, our prices have actually steadily declined, and we expect this to continue.

Financially, we are the soundest cryonics organization in existence.

We have a uniquely flexible and rapid system of emergency patient care based on universally available networks of mortuary assistance (and often medical assistance). This means that in the critical early stages, we can bring qualified professionals to you faster than any other system to you, and to travelers, vacationers, and members throughout most of the world. In particular, London-based F.A. Albin & Sons funeral directors are trained, practiced, equipped, and prepared to fly a team anywhere in Europe on short notice to help European CI members or tourists and business travelers.

And finally, we provide a comprehensive source of information here on CIs website. The site youre reading will lead you to everything you need to know about the subject of cryonics, and more. It offers you free information, free books, the latest news, hundreds of links to thousands of sources covering health, science, cutting-edge medicine, nanotechnology, financial help and resources, and supportive people and organizations. And if thats not enough? We personally will answer any question you might have about cryonics or the Cryonics Institute directly by email, or direct you to someone who can. In the world of cryonics, this is the source to visit, and the place to be.

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History Timeline - The Cryonics Institute

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The environment was something of a shift for Drake, who had spent the previous seven years as the medical response director of the Alcor Life Extension Foundation. Though it was the longtime leader in cryonics, Alcor was still a small nonprofit. It had been freezing the bodies and brains of its members, with the idea of one day bringing them back to life, since 1976.

The foundation, and cryonics in general, had long survived outside of mainstream acceptance. Typically shunned by the scientific community, cryonics is best known for its appearance in sci-fi films like 2001: A Space Odyssey. But its adherents have held on to a dream that at some point in the future, advances in medicine will allow for resuscitation and additional years on Earth. Over decades, small, tantalizing developments in related technology, as well as high-profile frozen test subjects like Ted Williams, have kept the hope alive. Today, nearly 200 dead patients are frozen in Alcors cryogenic chambers at temperatures of 196 C, including a handful of celebrities, who have paid tens of thousands of dollars for the goal of possible revival and ultimately reintegration into society.

But its the recent involvement of Yinfeng that signals something of a new era for cryonics. With impressive financial resources, government support, and scientific staff, its one of a handful of new labs focused on expanding the consumer appeal of cryonics and trying anew to bring credibility to the long-disputed theory of human reanimation. Just a year after Drake came on board as research director of the Shandong Yinfeng Life Science Research Institute, the subsidiary of the Yinfeng Biological Group overseeing the cryonics program, the institute performed its first cryopreservation. Its storage vats now hold about a dozen clients who are paying upwards of $200,000 to preserve the whole body.

Still, the field remains rooted in faith rather than any real evidence that it works. Its a hopeless aspiration that reveals an appalling ignorance of biology, says Clive Coen, a neuroscientist and professor at Kings College London.

Even if one day you could perfectly thaw a frozen human body, you would still just have a warm dead body on your hands.

The cryonics process typically goes something like this: Upon a persons death, a response team begins the process of cooling the corpse to a low temperature and performs cardiopulmonary support to sustain blood flow to the brain and organs. Then the body is moved to a cryonics facility, where an organ preservation solution is pumped through the veins before the body is submerged in liquid nitrogen. This process should commence within one hour of deaththe longer the wait, the greater the damage to the bodys cells. Then, once the frozen cadaver is ensconced in the cryogenic chamber, the hope of the dead begins.

Since its beginnings in the late 1960s, the field has attracted opprobrium from the scientific community, particularly its more respectable cousin cryobiologythe study of how freezing and low temperatures affect living organisms and biological materials. The Society for Cryobiology even banned its members from involvement in cryonics in the 1980s, with a former society president lambasting the field as closer to fraud than either faith or science.

In recent years, though, it has grabbed the attention of the libertarian techno-optimist crowd, mostly tech moguls dreaming of their own immortality. And a number of new startups are expanding the playing field. Tomorrow Biostasis in Berlin became the first cryonics company in Western Europe in 2019, for example, and in early 2022, Southern Cryonics opened a facility in Australia.

More researchers are open to longer-term, futuristic topics than there might have been 20 years ago or so, says Tomorrow Biostasis founder Emil Kendziorra.

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Why the sci-fi dream of cryonics never died - MIT Technology Review

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International Cryonics Museum has been chosen as the Sight of the Week by the editors of Roadside America.  Estes Park Trail-Gazette

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International Cryonics Museum has been chosen as the Sight of the Week by the editors of Roadside America. - Estes Park Trail-Gazette

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In case you missed it, link below to the CBS Sunday Morning segment about the Cryonics Museum  Estes Park Trail-Gazette

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In case you missed it, link below to the CBS Sunday Morning segment about the Cryonics Museum - Estes Park Trail-Gazette

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Testosterone replacement therapy may not increase risk of prostate cancer among men with hypogonadism  Medical Dialogues

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Testosterone replacement therapy may not increase risk of prostate cancer among men with hypogonadism - Medical Dialogues

Recommendation and review posted by Bethany Smith

Lesson 10: Could Science Really Extend The Human Lifespan?

/en/thenow/what-is-labgrown-meat/content/

Despite advances in medicine and nutrition, science has had difficulty extending the human lifespan beyond a certain age. Nevertheless, scientists are researching how to push beyond these limits and increase the duration and quality of a persons life. However, the big question remains: Could science really extend the human lifespan?

The science of life extension has one main goal, which is to prolong the human lifespan while maintaining youthful health. Since antiquity, people have pursued this goal through countless medicines, diets, and scientific procedures. But now, science has the knowledge to possibly make life extension a reality.

Life extension has been in the news lately because many entrepreneurs, especially within Silicon Valley, have publicly advocated and funded life extension research. Many of these advocates are incredibly optimistic because they see human aging not as inevitable but as an obstacle that will eventually be overcome.

Currently, there is no proven method of delaying or reversing the aging process. However, there are plenty of products available that will try to convince you otherwise.

Always be wary of any drug, food, or supplement that makes anti-aging claims. Also keep in mind that in the United States, the FDA only reviews supplements for safety, not for effectiveness. This means a supplement that claims to prevent aging could actually be useless. As for cosmetics that make anti-aging claims, these products are typically designed to hide the effects of aging, not reverse aging itself.

Essentially, if an anti-aging product seems too good to be true, it probably is.

Although life extension is being researched from numerous angles, genetic engineering has shown the most promise. Some methods involve replacing damaged cells with new ones, while others alter the mechanisms of DNA. In 2017, Harvard scientists discovered how to reverse aging in lab mice by changing how their DNA repairs itself. However, it's unclear if scientists will be able to duplicate this feat in humans.

While it can't delay aging itself, replacing worn organs could help people survive the more deteriorating effects of aging. Scientists have already grown kidneys and windpipes in laboratory settings using stem cells, and theyre researching how to create more complex organs like the heart and liver. There has also been significant progress with artificial organs, partially thanks to new technologies like 3D printing.

Currently, life extension is mostly research and educated guesses. Some experts believe we are not even close to breaking through the natural barriers of aging, while others believe some form of life extension will be widely available by the middle of the 21st century.

Almost everyone wants to live longer, healthier lives, but science has a long way to go before we overcome the natural limits of the human lifespan. Although life extension technology will continue to develop, major discoveries will take decades to realize, if they are realized at all.

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The Now: Could Science Really Extend The Human Lifespan? - GCFGlobal.org

Recommendation and review posted by Bethany Smith

In 2013, two biochemists published a paper proclaiming theyd discovered a potentially game-changing method of manipulating genes. CRISPR which sounds like a veggie-forward gastro pub won them each a Nobel Prize.

In the years since, CRISPR (or Clustered Regularly Interspaced Short Palindromic Repeats) has lived up to the hype. Its altered the global scientific landscape and raised questions about what kinds of revolutionary changes scientists and healthcare providers could and should pursue.

What if we could make foods allergy-free and crops drought-resistant? What if we could eliminate invasive species and protect against infectious diseases like malaria? What if we could revive extinct species? What if we could remove or repair mutations that cause inherited conditions? Or create custom immunotherapies to treat an individuals cancer?

The prospects are that exciting.

If your understanding of genetics starts and ends with high school biology or the (very fictional) Jurassic Park movies youre not alone. This stuff is complicated. Thats why we asked genomics and immunotherapy expert Timothy Chan, MD, PhD, to break CRISPR down for us, so we can better understand why, over a decade later, its still got researchers so excited.

Before we jump into CRISPR, lets start with the concept of gene editing.

Gene editing is the process of altering genetic material (DNA). That could mean changing a few individual genes or an entire sequence. Research has been ongoing for more than a decade thats looking at using gene editing on mutations that cause serious health conditions in people. The goal of this gene editing research is to eliminate or correct the mutation thats causing the health condition, or has the potential to cause one, such as certain cancers. In other research studies, gene editing is being explored so a mutation isnt passed down to children at birth.

For example, the U.S. Food and Drug Administration (FDA) approved a gene therapy in late 2022 that introduces a gene needed for blood clotting into people with hemophilia B. Its one of several cellular and gene therapy products currently in use today.

There are many different techniques and applications for gene editing. CRISPR is one approach to gene editing thats showing promise in ongoing clinical trials.

Now that were clear on what gene editing is, lets focus on a specific approach: CRISPR.

Clustered Regularly Interspaced Short Palindromic Repeats, otherwise known as CRISPR, was originally identified in bacteria, as a bacterial defense system, says Dr. Chan.

Thats right. Bacteria have immune systems, too.

CRISPR contains spacers sequences of DNA left over from unfriendly viruses or other entities as well as repeating sections of genetic material. Those sequences provide acquired immunity, and form the building blocks of the gene editing system or process. It creates a sort of blueprint that allows enzymes in genetic material to make changes to sequences of DNA in living cells. One of the best-known enzymes used for this purpose is called Cas9, which is why youll sometimes hear people talk about CRISPR-Cas9.

Over the years, people have discovered that specific enzymes that allow CRISPR to work Cas9 is one of them.But there are other ones, and they can be tailored to target sequences of interest in the DNA for specific cuts to be made, Dr. Chan explains.

You can think of the underlying mechanism of CRISPR gene editing as being similar to the way magnetic shapes are drawn to each other or the way Lego blocks fit together.

The segments in CRISPR are transcribed into RNA. This RNA includes a guide sequence, which is a match to existing DNA in a persons body.

That guide sequence can be tailored to whatever you want, Dr. Chan says. And as a result, you can make specific alterations or mutations in a part of the genome that you are targeting with a high degree of accuracy.

Along for the ride with this guide sequence is an enzyme like Cas9.

When the guide sequence and enzyme find the desired DNA to edit, the enzyme can then get down to business. It attaches itself to this DNA and makes changes, whether thats a cut or alteration.

CRISPR technology has come a long way, Dr. Chan says. The first generation of CRISPR was a great way to inactivate genes. It only made a break in genes. Then, the DNA would get filled up with natural repair enzymes.

But new versions of CRISPR like CRISPR prime or CRISPR HD are more advanced.

These can allow actual replacements to occur, Dr. Chan continues. You can even very accurately replace one sequence one of the letters in the genome with another letter. And you can make specific mutations.

CRISPRs ability to make very specific, very small cuts has the potential to transform how healthcare providers can address certain genetic diseases.

Dr. Chan is optimistic about the future of CRISPR based on the success of ongoing clinical trials in human subjects. For any type of genetic diseases caused by a single mutated gene, you can use CRISPR to mutate it and make it normal. Thats why its useful. Its a way for us to change errors in the genome.

Right now, CRISPR is geared toward correcting a single change in genes, he adds. While combinations may be possible in the future, were just not there yet.

While gene editing is already in use, CRISPR is still in the clinical trials phase, Dr. Chan says. Its used all the time in research laboratories and industries, he notes. Many clinical trials are testing CRISPR in the setting of genetic diseases and cancer.

Interestingly, CRISPR can be used to detect certain diseases. The best-known example is the Sherlock CRISPR SARS-CoV-2 Kit: A COVID-19 test that received emergency use authorization (EAU) from the FDA in 2020.

But theres no FDA-approved CRISPR therapy right now. The clinical trials are ongoing, he says.

These include trials looking at CRISPR to correct genetic diseases such as cystic fibrosis, Huntingtons disease and muscular dystrophy.

Dr. Chan adds that there are also major clinical trials in process for blood disorders, where CRISPR is being used to correct the gene alteration that causes the condition. As one example, he cites a promising trial looking at CRISPR-Cas9 gene editing for sickle cell disease and -thalassemia, written about in an early 2021 issue of the New England Journal of Medicine. -thalassemia is an inherited blood disorder that impacts the bodys ability to create hemoglobin an iron-dense protein that serves as the primary ingredient in red blood cells.

There are also clinical trials looking to see if CRISPR can be used to treat certain cancers. Dr. Chan notes that chimeric antigen receptor (CAR)T-cell therapy is one of the first gene therapies approved for leukemias. Current research is looking at whether CRISPR technology can make this treatment even more effective.

In CAR T-cell therapy, you take out T-cells from someone and put in a receptor a new way for these cells to target something on cancer cells and then put these cells back in the patient, he explains. Researchers are running trials now where they use CRISPR to alter those T-cells to make them even more active.

CRISPR therapies can take on many different forms. CRISPR has been inserted directly into the body before. It was famously injected into the eyes of seven people with a rare hereditary blindness disorder in 2020, two of whom later told NPR that they regained some ability to see colors. There are human trials in process right now that deliver CRISPR through gels and creams, through food or drink, skin grafts or injections. Ex-vivo delivery is also common: Thats when CRISPR is used to modify a cell outside the body. The cells are then re-inserted into the body using a harmless virus.

The results have been promising so far. I do believe in the next three to five years possibly even sooner were going to see approval to treat some diseases, Dr. Chan states.

With any type of CRISPR therapy, Dr. Chan says theres a risk of getting off-target effects or unexpected side effects.

Whenever youre altering something as fundamental as DNA, you just dont know what might happen he explains. Theres always a chance for the unexpected. You can potentially have effects on your DNA that were not intended.

At the moment, he doesnt have any specific examples of what these effects might be and he notes that existing research suggests the risk is pretty low. Still, data from future research might tell a different story.

Dr. Chan nevertheless sees a lot of potential for CRISPR in the coming years.

The field is moving very quickly, he says. Were seeing continual improvement of the actual CRISPR tools being used.

Its getting more accurate and more flexible in terms of what you can do. There are various engineered modified variants of CRISPR now that are allowing very specific, very accurate changes with fewer off-target effects. So, I think the future is very bright.

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What Is CRISPR Gene Editing and How Does It Work?

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Stephanie is professional, personable, and a joy to be around. She puts you right at ease. Explains all procedures. She is very knowledgeable in all aspects of her profession.

(Injectable and hormone patient TS from Awendaw)

I really bonded with Stephanie! She explained things so well. I am looking forward to seeing her again

(Hormone patient- MS from James Island)

Stephanies the best!

(hormone and injectable patient BR from Mount Pleasant)

Stephanie is a life changer! I know my body well, and going through changes she makes me feel like I am not loosing my mind. Going to my regular OB doctor in regards to hormonal issues I have been told that what I am going through is not pre- menopausal. Wrong!!! Stephanie has me set up on hormone replacement therapy that has been a game changer. I feel great, I have a ton of energy and I most importantly I feel balanced all day long. Her knowledge and expertise is top notch! I highly recommend her. Not only is she knowledgeable, she goes the extra mile to explain things, she also gives you options to what you as a patient are comfortable with. I love that she supports the route (you as a patient) decides to do. I highly recommend Stephanie at Charleston Hormone Replacement and Aesthetic Services.

Hormone Patient

Stephanie Donovan is the absolute best at what she does. She takes her time to review your labs and to talk about any aesthetic procedures in detail so that you feel completely comfortable with what your goal is. Shes knowledgeable about the latest technology and knows how to customize the right solution just for you. I trust my face, lips, and everything with her!

Hormone Patient

Stephanie is amazing! Love the new office. She has helped me improve my quality of life so much I cant even explain how good I feel all the time now. Super friendly, professional, and always caring and super helpful. Cant recommend her enough!

Hormone Patient

Stephanie is amazing at what she does! She has changed my life with educating me on the importance of hormone therapy and has provided excellent care. She has a heart for what she does and truly cares about her patients.

Hormone Patient

Cannot say enough good things about my experience with Stephanie! She thoroughly takes the time to listen and care for her patients. The best in Charleston for injectables and hormones!

Hormone and Cosmetic Patient

So happy I finally found Steph again!!! She is the BEST in Charleston. Definitely give her a try for all your aesthetic needs as well womens health!! Hormone guru!!! Highly recommend !!!!

Aesthetic patient from Isle of Palms, SC

Everyone should get their levels checked at one or more points in life to optimize our health and general feeling of wellness and this is a great place to do it! The initial consult was very attentive and detailed. Stephanie spent a lot of time explaining my results so that I felt confident in the drivers of my symptoms and next steps I needed to take. Definitely recommend. 🙂

-Hormone Patient from Charleston, SC

I went today to have a pellet as Im visiting from out of town and it was time. I was nervous because it was my first time here and I wanted to make sure it would be pain-free. Stephanie was fantastic. Put my mind at ease. It was not painful at all and she was wonderful to deal with!

-Hormone Patient from Delray Beach, FL

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Home - CHS Hormone

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My teenage son was indoctrinated by the trans cult from the classroom to the clinic: Mother left 'grieving' fo  Daily Mail

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My teenage son was indoctrinated by the trans cult from the classroom to the clinic: Mother left 'grieving' fo - Daily Mail

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Blood tests to measure hormone levels

An evaluation usually begins by measuring blood levels of the hormones that the pituitary gland produces (typically, thyroid-stimulating hormone, prolactin, luteinizing hormone, and follicle-stimulating hormone) and at the same time measuring levels of the hormone produced by the target organs (typically, thyroid hormone, testosterone in men, and estrogen in women).

For example, a person with hypothyroidism due to failure of the pituitary gland has low levels of thyroid hormone and low or inappropriately normal levels of thyroid-stimulating hormone, which is produced by the pituitary gland. In contrast, a person with hypothyroidism due to failure of the thyroid gland itself has low levels of thyroid hormone and high levels of thyroid-stimulating hormone.

Growth hormone production by the pituitary is difficult to evaluate because no single blood level accurately reflects it. The body usually produces growth hormone in several bursts a night, and the hormone is quickly used. Thus, the blood level at any given moment does not indicate whether production is normal over the course of a day. Instead, doctors measure the levels of insulin-like growth factor 1 (IGF-1) in the blood. Production of IGF-1 is controlled by growth hormone, and the level of IGF-1 tends to change slowly in proportion to the overall amount of growth hormone produced by the pituitary. In infants and young children, doctors may instead measure levels of a similar substance, IGF-binding protein type 3. Measurement of IGF-1 is not sufficient to make the diagnosis of growth hormone deficiency in adults, because people with normal levels may still have growth hormone deficiency. In many cases, a stimulation test to try to make the pituitary secrete growth hormone is used.

Because the levels of luteinizing hormone and follicle-stimulating hormone fluctuate with the menstrual cycle, their measurement in women may be difficult to interpret. However, in postmenopausal women who are not taking estrogen, luteinizing hormone and follicle-stimulating hormone levels normally are high. When they are found to be low, this can be an indication of pituitary damage or insufficiency of other hormones.

Production of ACTH is usually assessed by measuring the levels of its target hormone (cortisol) in response to stimuli, such as an injection of synthetic ACTH (ACTH stimulation test) or a low level of sugar in the blood after an insulin injection (insulin tolerance test). If the level of cortisol does not change and the level of ACTH in the blood is normal or low, a deficiency of ACTH production is confirmed.

Read more here:
Hypopituitarism - Hormonal and Metabolic Disorders - Merck Manuals ...

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In 2013, two biochemists published a paper proclaiming theyd discovered a potentially game-changing method of manipulating genes. CRISPR which sounds like a veggie-forward gastro pub won them each a Nobel Prize.

Cleveland Clinic is a non-profit academic medical center. Advertising on our site helps support our mission. We do not endorse non-Cleveland Clinic products or services. Policy

In the years since, CRISPR (or Clustered Regularly Interspaced Short Palindromic Repeats) has lived up to the hype. Its altered the global scientific landscape and raised questions about what kinds of revolutionary changes scientists and healthcare providers could and should pursue.

What if we could make foods allergy-free and crops drought-resistant? What if we could eliminate invasive species and protect against infectious diseases like malaria? What if we could revive extinct species? What if we could remove or repair mutations that cause inherited conditions? Or create custom immunotherapies to treat an individuals cancer?

The prospects are that exciting.

If your understanding of genetics starts and ends with high school biology or the (very fictional) Jurassic Park movies youre not alone. This stuff is complicated. Thats why we asked genomics and immunotherapy expert Timothy Chan, MD, PhD, to break CRISPR down for us, so we can better understand why, over a decade later, its still got researchers so excited.

Before we jump into CRISPR, lets start with the concept of gene editing.

Gene editing is the process of altering genetic material (DNA). That could mean changing a few individual genes or an entire sequence. Research has been ongoing for more than a decade thats looking at using gene editing on mutations that cause serious health conditions in people. The goal of this gene editing research is to eliminate or correct the mutation thats causing the health condition, or has the potential to cause one, such as certain cancers. In other research studies, gene editing is being explored so a mutation isnt passed down to children at birth.

For example, the U.S. Food and Drug Administration (FDA) approved a gene therapy in late 2022 that introduces a gene needed for blood clotting into people with hemophilia B. Its one of several cellular and gene therapy products currently in use today.

There are many different techniques and applications for gene editing. CRISPR is one approach to gene editing thats showing promise in ongoing clinical trials.

Now that were clear on what gene editing is, lets focus on a specific approach: CRISPR.

Clustered Regularly Interspaced Short Palindromic Repeats, otherwise known as CRISPR, was originally identified in bacteria, as a bacterial defense system, says Dr. Chan.

Thats right. Bacteria have immune systems, too.

CRISPR contains spacers sequences of DNA left over from unfriendly viruses or other entities as well as repeating sections of genetic material. Those sequences provide acquired immunity, and form the building blocks of the gene editing system or process. It creates a sort of blueprint that allows enzymes in genetic material to make changes to sequences of DNA in living cells. One of the best-known enzymes used for this purpose is called Cas9, which is why youll sometimes hear people talk about CRISPR-Cas9.

Over the years, people have discovered that specific enzymes that allow CRISPR to work Cas9 is one of them.But there are other ones, and they can be tailored to target sequences of interest in the DNA for specific cuts to be made, Dr. Chan explains.

You can think of the underlying mechanism of CRISPR gene editing as being similar to the way magnetic shapes are drawn to each other or the way Lego blocks fit together.

The segments in CRISPR are transcribed into RNA. This RNA includes a guide sequence, which is a match to existing DNA in a persons body.

That guide sequence can be tailored to whatever you want, Dr. Chan says. And as a result, you can make specific alterations or mutations in a part of the genome that you are targeting with a high degree of accuracy.

Along for the ride with this guide sequence is an enzyme like Cas9.

When the guide sequence and enzyme find the desired DNA to edit, the enzyme can then get down to business. It attaches itself to this DNA and makes changes, whether thats a cut or alteration.

CRISPR technology has come a long way, Dr. Chan says. The first generation of CRISPR was a great way to inactivate genes. It only made a break in genes. Then, the DNA would get filled up with natural repair enzymes.

But new versions of CRISPR like CRISPR prime or CRISPR HD are more advanced.

These can allow actual replacements to occur, Dr. Chan continues. You can even very accurately replace one sequence one of the letters in the genome with another letter. And you can make specific mutations.

CRISPRs ability to make very specific, very small cuts has the potential to transform how healthcare providers can address certain genetic diseases.

Dr. Chan is optimistic about the future of CRISPR based on the success of ongoing clinical trials in human subjects. For any type of genetic diseases caused by a single mutated gene, you can use CRISPR to mutate it and make it normal. Thats why its useful. Its a way for us to change errors in the genome.

Right now, CRISPR is geared toward correcting a single change in genes, he adds. While combinations may be possible in the future, were just not there yet.

While gene editing is already in use, CRISPR is still in the clinical trials phase, Dr. Chan says. Its used all the time in research laboratories and industries, he notes. Many clinical trials are testing CRISPR in the setting of genetic diseases and cancer.

Interestingly, CRISPR can be used to detect certain diseases. The best-known example is the Sherlock CRISPR SARS-CoV-2 Kit: A COVID-19 test that received emergency use authorization (EAU) from the FDA in 2020.

But theres no FDA-approved CRISPR therapy right now. The clinical trials are ongoing, he says.

These include trials looking at CRISPR to correct genetic diseases such as cystic fibrosis, Huntingtons disease and muscular dystrophy.

Dr. Chan adds that there are also major clinical trials in process for blood disorders, where CRISPR is being used to correct the gene alteration that causes the condition. As one example, he cites a promising trial looking at CRISPR-Cas9 gene editing for sickle cell disease and -thalassemia, written about in an early 2021 issue of the New England Journal of Medicine. -thalassemia is an inherited blood disorder that impacts the bodys ability to create hemoglobin an iron-dense protein that serves as the primary ingredient in red blood cells.

There are also clinical trials looking to see if CRISPR can be used to treat certain cancers. Dr. Chan notes that chimeric antigen receptor (CAR)T-cell therapy is one of the first gene therapies approved for leukemias. Current research is looking at whether CRISPR technology can make this treatment even more effective.

In CAR T-cell therapy, you take out T-cells from someone and put in a receptor a new way for these cells to target something on cancer cells and then put these cells back in the patient, he explains. Researchers are running trials now where they use CRISPR to alter those T-cells to make them even more active.

CRISPR therapies can take on many different forms. CRISPR has been inserted directly into the body before. It was famously injected into the eyes of seven people with a rare hereditary blindness disorder in 2020, two of whom later told NPR that they regained some ability to see colors. There are human trials in process right now that deliver CRISPR through gels and creams, through food or drink, skin grafts or injections. Ex-vivo delivery is also common: Thats when CRISPR is used to modify a cell outside the body. The cells are then re-inserted into the body using a harmless virus.

The results have been promising so far. I do believe in the next three to five years possibly even sooner were going to see approval to treat some diseases, Dr. Chan states.

With any type of CRISPR therapy, Dr. Chan says theres a risk of getting off-target effects or unexpected side effects.

Whenever youre altering something as fundamental as DNA, you just dont know what might happen he explains. Theres always a chance for the unexpected. You can potentially have effects on your DNA that were not intended.

At the moment, he doesnt have any specific examples of what these effects might be and he notes that existing research suggests the risk is pretty low. Still, data from future research might tell a different story.

Dr. Chan nevertheless sees a lot of potential for CRISPR in the coming years.

The field is moving very quickly, he says. Were seeing continual improvement of the actual CRISPR tools being used.

Its getting more accurate and more flexible in terms of what you can do. There are various engineered modified variants of CRISPR now that are allowing very specific, very accurate changes with fewer off-target effects. So, I think the future is very bright.

Continue reading here:
The Basics of CRISPR Gene Editing - Cleveland Clinic Health Essentials

Recommendation and review posted by Bethany Smith

Abstract

Clustered regularly interspaced short palindromic repeat (CRISPR) and their associated protein (Cas-9) is the most effective, efficient, and accurate method of genome editing tool in all living cells and utilized in many applied disciplines. Guide RNA (gRNA) and CRISPR-associated (Cas-9) proteins are the two essential components in CRISPR/Cas-9 system. The mechanism of CRISPR/Cas-9 genome editing contains three steps, recognition, cleavage, and repair. The designed sgRNA recognizes the target sequence in the gene of interest through a complementary base pair. While the Cas-9 nuclease makes double-stranded breaks at a site 3 base pair upstream to protospacer adjacent motif, then the double-stranded break is repaired by either non-homologous end joining or homology-directed repair cellular mechanisms. The CRISPR/Cas-9 genome-editing tool has a wide number of applications in many areas including medicine, agriculture, and biotechnology. In agriculture, it could help in the design of new grains to improve their nutritional value. In medicine, it is being investigated for cancers, HIV, and gene therapy such as sickle cell disease, cystic fibrosis, and Duchenne muscular dystrophy. The technology is also being utilized in the regulation of specific genes through the advanced modification of Cas-9 protein. However, immunogenicity, effective delivery systems, off-target effect, and ethical issues have been the major barriers to extend the technology in clinical applications. Although CRISPR/Cas-9 becomes a new era in molecular biology and has countless roles ranging from basic molecular researches to clinical applications, there are still challenges to rub in the practical applications and various improvements are needed to overcome obstacles.

Keywords: CRISPR, Cas-9, sgRNA, gene-editing, mechanism, applications

Genome editing is a type of genetic engineering in which DNA is deliberately inserted, removed, or modified in living cells.1 The name CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) refers to the unique organization of short, partially repeated DNA sequences found in the genomes of prokaryotes. CRISPR and its associated protein (Cas-9) is a method of adaptive immunity in prokaryotes to defend themselves against viruses or bacteriophages.2 Japanese scientist Ishino and his team accidentally found unusual repetitive palindromic DNA sequences interrupted by spacers in Escherichia coli while analyzing a gene for alkaline phosphatase first discovered CRISPR in 1987. However, they did not ascertain its biological function. In 1990, Francisco Mojica identifies similar sequences in other prokaryotes and he named CRISPR, yet the functions of these sequences were a mystery.3 Later on in 2007, a CRISPR was experimentally conferred as a key element in the adaptive immune system of prokaryotes against viruses. During the adaptation process, bacterial cells become immunized by the insertion of short fragments of viral DNA (spacers) into a genomic region called the CRISPR array. Hence, spacers serve as a genetic memory of previous viral infections.4 The CRISPR defense mechanism protects bacteria from repeated viral attacks via three basic stages: adaptation (spacer acquisition), crRNA synthesis (expression), and target interference. CRISPR loci are an array of short repeated sequences found in chromosomal or plasmid DNA of prokaryotes. Cas gene is usually found adjacent to CRISPR that codes for nuclease protein (Cas protein) responsible to destroy or cleave viral nucleic acid.5

Before the discovery of CRISPR/Cas-9, scientists were relied on two gene-editing techniques using restriction enzymes, zinc finger nucleases (ZFN) and Transcription activator-like effector nucleases (TALENs).6 ZFN has a zinc finger DNA binding domain used to bind a specific target DNA sequence and a restriction endonuclease domain used to cleave the DNA at the target site. TALENs are also composed of DNA binding domain and restriction domain like ZFN but their DNA binding domain has more potential target sequence than the ZFN gene-editing tool. In both cases, the difficulty of protein engineering, being expensive, and time-consuming were the major challenges for researchers and manufacturers.6,7 The development of a reliable and efficient method of a gene-editing tool in living cells has been a long-standing goal for biomedical researchers. After figuring out the CRISPR mechanism in prokaryotes, scientists understood that it could have beneficial use in humans, plants, and other microbes. It was in 2012 that Doudna, J, and Charpentier, E discovered CRISPR/Cas-9 could be used to edit any desired DNA by just providing the right template.8 Since then, CRISPR/Cas-9 becomes the most effective, efficient, and accurate method of genome editing tool in all living cells and utilized in many applied disciplines.9 Thus, this review aims to discuss the mechanisms of genome editing mediated by CRISPR/Cas-9 and to highlight its recent applications as one of the most important scientific discoveries of this century, as well as the current barriers to the transformation of this technology.

Based on the structure and functions of Cas-proteins, CRISPR/Cas system can be divided into Class I (type I, III, and IV) and Class II (type II, V, and VI). The class I systems consist of multi-subunit Cas-protein complexes, while the class II systems utilize a single Cas-protein. Since the structure of type II CRISPR/Cas-9 is relatively simple, it has been well studied and extensively used in genetic engineering.10 Guide RNA (gRNA) and CRISPR-associated (Cas-9) proteins are the two essential components in CRISPR/Cas-9 system. The Cas-9 protein, the first Cas protein used in genome editing was extracted from Streptococcus pyogenes (SpCas-9). It is a large (1368 amino acids) multi-domain DNA endonuclease responsible for cleaving the target DNA to form a double-stranded break and is called a genetic scissor.11 Cas-9 consists of two regions, called the recognition (REC) lobe and the nuclease (NUC) lobe. The REC lobe consists of REC1 and REC2 domains responsible for binding guide RNA, whereas the NUC lobe is composed of RuvC, HNH, and Protospacer Adjacent Motif (PAM) interacting domains. The RuvC and HNH domains are used to cut each single-stranded DNA, while PAM interacting domain confers PAM specificity and is responsible for initiating binding to target DNA.12 Guide RNA is made up of two parts, CRISPR RNA (crRNA) and trans-activating CRISPR RNA (tracrRNA). The crRNA is an 1820 base pair in length that specifies the target DNA by pairing with the target sequence, whereas tracrRNA is a long stretch of loops that serve as a binding scaffold for Cas-9 nuclease. In prokaryotes, the guide RNA is used to target viral DNA, but in the gene-editing tool, it can be synthetically designed by combining crRNA and tracrRNA to form a single guide RNA (sgRNA) in order to target almost any gene sequence supposed to be edited.11

The mechanism of CRISPR/Cas-9 genome editing can be generally divided into three steps: recognition, cleavage, and repair.13 The designed sgRNA directs Cas-9 and recognizes the target sequence in the gene of interest through its 5crRNA complementary base pair component. The Cas-9 protein remains inactive in the absence of sgRNA. The Cas-9 nuclease makes double-stranded breaks (DSBs) at a site 3 base pair upstream to PAM.14 PAM sequence is a short (25 base-pair length) conserved DNA sequence downstream to the cut site and its size varies depending on the bacterial species. The most commonly used nuclease in the genome-editing tool, Cas-9 protein recognizes the PAM sequence at 5-NGG-3 (N can be any nucleotide base). Once Cas-9 has found a target site with the appropriate PAM, it triggers local DNA melting followed by the formation of RNA-DNA hybrid, but the mechanism of how Cas-9 enzyme melts target DNA sequence was not clearly understood yet. Then, the Cas-9 protein is activated for DNA cleavage. HNH domain cleaves the complementary strand, while the RuvC domain cleaves the non-complementary strand of target DNA to produce predominantly blunt-ended DSBs. Finally, the DSB is repaired by the host cellular machinery.11,15

Non-homologous end joining (NHEJ), and homology-directed repair (HDR) pathways are the two mechanisms to repair DSBs created by Cas-9 protein in CRISPR/Cas-9 mechanism.16 NHEJ facilitates the repair of DSBs by joining DNA fragments through an enzymatic process in the absence of exogenous homologous DNA and is active in all phases of the cell cycle. It is the predominant and efficient cellular repair mechanism that is most active in the cells, but it is an error-prone mechanism that may result in small random insertion or deletion (indels) at the cleavage site leading to the generation of frameshift mutation or premature stop codon.17 HDR is highly precise and requires the use of a homologous DNA template. It is most active in the late S and G2 phases of the cell cycle. In CRISPR-gene editing, HDR requires a large amount of donor (exogenous) DNA templates containing a sequence of interest. HDR executes the precise gene insertion or replacement by adding a donor DNA template with sequence homology at the predicted DSB site.16,17

In just a few years of its discovery, the CRISPR/Cas-9 genome editing tool has already being explored for a wide number of applications and had a massive impact on the world in many areas including medicine, agriculture, and biotechnology. In the future, researchers hope that this technology will continue to advance for treating and curing diseases, develop more nutritious crops, and eradicating infectious diseases.18 Highlights for some of the recent CRISPR/Cas-9 applications and clinical trials being investigated are discussed below.

More than 6000 genetic disorders have been known so far. But the majority of the diseases lack effective treatment strategies.19 Gene therapy is the process of replacing the defective gene with exogenous DNA and editing the mutated gene at its native location. It is the latest development in the revolution of medical biotechnology. From 1998 to August 2019, 22 gene therapies including the novel CRISPR/Cas-9 have been approved for the treatment of human diseases.20

Since its discovery in 2012, CRISPR/Cas-9 gene editing has held the promise of curing most of the known genetic diseases such as sickle cell disease, -thalassemia, cystic fibrosis, and muscular dystrophy.21,22 CRISPR/Cas-9 for targeted sickle cell disease (SCD) therapy and -thalassemia have been also applied in clinical trials.23 SCD is an autosomal recessive genetic disease of red blood cells, which occurs due to point mutation in the -globin chain of hemoglobin leading to sickle hemoglobin (HbS). During the deoxygenation process, HbS polymerization leads to severe clinical complications like hemolytic anemia.24 Either direct repairing the gene of hemoglobin S or boosting fetal -globin are the two main approaches that CRISPR/Cas-9 is being used to treat SCD.25 However, the most common method used in a clinical trial is based on the approach of boosting fetal hemoglobin. First bone marrow cells are removed from patients and the gene that turns off fetal hemoglobin production, called B-cell Lymphoma 11A (BCL11A) is disabled with CRISPR/Cas-9. Then, the gene-edited cells are infused back into the body.26 BCL11A is a 200 base pair gene found on chromosome 2 and its product is responsible to switch -globin into the -globin chain by repressing -globin gene expression.27 Once this gene is disabled using CRISPR/Cas-9, the production of fetal hemoglobin containing -globin in the red blood cells will increase, thereby alleviating the severity and manifestations of SCD.28

Scientists have been also investigating CRISPR/Cas-9 for the treatment of cystic fibrosis. The genetic mutation of the cystic fibrosis transmembrane conductance regulator (CFTR) gene decreases the structural stability and function of CFTR protein leading to cystic fibrosis.29 CFTR protein is an anion channel protein regulated by protein kinase-A, located at the apical surface of epithelial cells of the lung, intestine, pancreas, and reproductive tract.30 Although there is no cure for cystic fibrosis, symptom-based therapies (such as antibiotics, bronchodilators, and mucus thinning medications) and CFTR modulating drugs have become the first-line treatments to relieve symptoms and reduce the risk of complications.31 Currently, gene manipulation technologies and molecular targets are also being explored. The use of CRISPR/Cas-9 technology for genome editing has great potential, although it is in the early stages of development.32 In 2013, researchers culture intestinal stem cells from two cystic fibrosis patients and corrected the mutation at the CFTR locus resulting in the expression of the correct gene and full function of the protein. Since then, the potential utility of the application of CRISPR/Cas-9 for cystic fibrosis was established.33 Furthermore, Duchenne muscular dystrophy (DMD), which is caused by a mutation in the dystrophin gene and characterized by muscle weakness, has been successfully corrected by CRISPR/Cas-9 in patient-induced pluripotent stem cells.34 Despite considerable efforts, the treatment available for DMD remains supportive rather than curative. Currently, several therapeutic approaches (gene therapy, cell therapy, and exon skipping) have been investigated to restore the expression of dystrophin in DMD muscles.35,36 Deletion/excision of intragenic DNA and removing the duplicated exon by CRISPR/Cas-9 are the new and promising approaches in correcting the DMD gene, which restores the expression of dystrophin protein.37

Moreover, the latest researches show that the CRISPR/Cas-mediated single-base editing and prime editing systems can directly install mutations in cellular DNA without the need for a donor template. The CRISPR/Cas-base editor and prime editor system do not produce DSB, which reduces the possibility of indels that are different from conventional Cas-9.38 So far, two types of base editors have been developed: cytosine base editor (CBE) and adenine base editor (ABE).39 The CBE is a type of base editor composed of cytidine deaminase fused with catalytically deficient or dead Cas-9 (dCas-9). It is one of the novel gene therapy strategies that can produce precise base changes from cytidine (C) to thymidine (T).40 However, the target range of the CBE base editor is still restricted by PAM sequences containing G, T, or A bases. Recently, a more advanced fidelity and efficiency base editor called nNme2-CBE (discovered from Neisseria meningitides) with expanded PAM compatibility for cytidine dinucleotide has been developed in both human cells and rabbits embryos.41 The ABE uses adenosine deaminase fused to dCas-9 to correct the base-pair change from adenosine (A) to guanosine (G).38 Overall, single-base editing through the fusion of dCas-9 to cytidine deaminase or adenosine deaminase is a safe and efficient method to edit point mutations. But both base editors can only fix four-transition mutations (purine to purine or pyrimidine to pyrimidine).42 To overcome this shortcoming, the most recent member of the CRISPR genome editing toolkit called Prime Editor (PE) has been developed to extend the scope of DNA editing beyond the four types of transition mutations.43 PE contains Cas-9 nickase fused with engineered reverse transcriptase and multifunctional primer editing guide RNA (pegRNA). The pegRNA recognizes the target nucleotide sequence; the Cas-9 nickase cuts the non-complementary strand of DNA three bases upstream from the PAM site, exposing a 3-OH nick of genomic DNA. The reverse transcriptase then extends the 3 nick by copying the edit sequence of pegRNA. Hence, PE not only corrects all 12 possible base-to-base transitions, and transversion mutations but also small insertion and deletion mutations in genetic disorders.44

The first CRISPR-based therapy in the human trial was conducted to treat patients with refractory lung cancer. Researchers first extract T-cells from three patients blood and they engineered them in the lab through CRISPR/Cas-9 to delete genes (TRAC, TRBC, and PD-1) that would interfere to fight cancer cells. Then, they infused the modified T-cells back into the patients. The modified T-cells can target specific antigens and kill cancer cells. Finally, no side effects were observed and engineered T-cells can be detected up to 9 months of post-infusion.45 CRISPR/Cas-9 gene-editing technology could also be used to treat infectious diseases caused by microorganisms.46 One focus area for the researchers is treating HIV, the virus that leads to AIDS. In May 2017, a team of researchers from Temple University demonstrated that HIV-1 replication can be completely shut down and the virus eliminated from infected cells through excision of HIV-1 genome using CRISPR/Cas-9 in animal models.47 In addition to the approach of targeting the HIV-genome, CRISPR/Cas-9 technology can also be used to block HIV entry into host cells by editing chemokine co-receptor type-5 (CCR5) genes in the host cells. For instance, an in vitro trial conducted in China reported that genome editing of CCR5 by CRISPR/Cas-9 showed no evidence of toxicity (infection) on cells and they concluded that edited cells could effectively be protected from HIV infection than unmodified cells.48

As the world population continues to grow, the risk of shortage in agricultural resources is real. Hence, there is a need for new technologies for increasing and improving natural food production. CRISPR/Cas-9 is an existing addition to the field since it has been used to genetically modify foods to improve their nutritional value, increase their shelf life, make them drought-tolerant, and enhance disease resistance.18 There are generally three ways that CRISPR is solving the worlds food crisis. It can restore food supplies, help plants to survive in hostile conditions, and could improve the overall health of the plants.49

Beyond genome editing activity, CRISPR/Cas-9 can be used to artificially regulate (activate or repress) a certain target of a gene through advanced modification of Cas-9 protein.15 Researchers had performed an advanced modified Cas-9 endonuclease called dCas-9 nuclease by inactivating its HNH and RuvC domains. The dCas-9 nuclease lacks DNA cleavage activity, but its DNA binding activity is not affected. Then, transcriptional activators or inhibitors can be fused with dCas-9 to form the CRISPR/dCas-9 complex. Therefore, catalytically inactive dCas-9 can be used to activate (CRISPRa) or silence (CRISPRi) the expression of a specific gene of interest.50 Moreover, the CRISPR/dCas-9 can be also used to visualize and pinpoint where specifically the gene of interest is located inside the cell (subcellular localization) by fusing a marker such as Green Fluorescent Proteins (GFP) with dCas-9 enzyme. This enables site-specific labeling and imaging of endogenous loci in living cells for further utilization.51

Despite its great promise as a genome-editing system CRISPR/Cas-9 technology had hampered by several challenges that should be addressed during the process of application. Immunogenicity, lack of a safe and efficient delivery system to the target, off-target effect, and ethical issues have been the major barriers to extend the technology in clinical applications.52 Since the components of the CRISPR/Cas-9 system are derived from bacteria, host immunity can elicit an immune response against these components. Researchers also found that there were both pre-existing humoral (anti-Cas-9 antibody) and cellular (anti-Cas-9 T cells) immune responses to Cas-9 protein in healthy humans. Therefore, how to detect and reduce the immunogenicity of Cas-9 protein is still one of the most important challenges in the clinical trial of the system.53

Safe and effective delivery of the components into the cell is essential in CRISPR/Cas-9 gene editing. Currently, there are three methods of delivering the CRISPR/Cas-9 complex into cells, physical, chemical, and viral vectors. Non-viral (physical and chemical) methods are more suitable for ex vivo CRISPR/Cas-9-based gene editing therapy.54 The physical methods of delivering CRISPR/Cas-9 can include electroporation, microinjection, hydrodynamic injection, and so on. Electroporation applies a strong electric field to the cell membrane to temporarily increase the permeability of the membrane, allowing the CRISPR/Cas-9 complex to enter the cytoplasm of the target cell. However, the main limitation of this method is that it causes significant cell death.55 Microinjection involves injecting the CRISPR/Cas-9 complex directly into cells at the microscopic level for rapid gene editing of a single cell. Nevertheless, this method also has several disadvantages such as cell damage, which is technically challenging and is only suitable for a limited number of cells.56 The hydrodynamic injection is the rapid injection of a large amount of high-pressure liquid into the bloodstream of animals, usually using the tail vein of mice. Although this method is simple, fast, efficient, and versatile, it has not yet been used in clinical applications due to possible complications.57 The chemical methods of CRISPR/Cas-9 delivery involves lipid and polymer-based nanoparticles.58 Lipid nanoparticles/liposomes are spherical structures composed of lipid bilayers membrane and are synthesized in aqueous solutions using Lipofectamine-based reagents. The positively charged liposomes encapsulated with negatively charged nucleic acids thereby facilitate the fusion of the complex across the cell membrane into cells.59 Polymeric nanoparticles, such as polyethyleneimine and poly-L-lysine, are the most widely used carriers of CRISPR/Cas-9 components. Like lipid nanoparticles, polymer-based nanoparticles can also transverse the complex in the membrane through endocytosis.60

Viral vectors are the natural experts for in vivo CRISPR/Cas-9 delivery.61 Vectors, such as adenoviral vectors (AVs), adeno-associated viruses (AAVs), and lentivirus vectors (LVs) are currently being widely used as delivery methods due to their higher delivery efficiency relative to physical and chemical methods. Among them, AAVs are the most commonly used vectors due to their low immunogenicity and non-integration into the host cell genome compared to other viral vectors.62 However, the limited virus cloning capacity and the large size of the Cas-9 protein remain a major problem. One strategy to tackle this hurdle is to package sgRNA and Cas-9 into separate AAVs and then co-transfect them into cells. Recent methods employ a smaller strain of Cas-9 from Staphylococcus aureus (SaCas-9) instead of the more commonly used SpCas-9 to allow packaging of sgRNA and Cas-9 in the same AAVs.54,61 Lately, the development of extracellular vesicles (EVs), for the in vivo delivery of CRISPR/Cas-9 to avoid some of the limitations of viral and non-viral methods has shown a great potential.63

The designed sgRNA will mismatch to the non-target DNA and can result in nonspecific, unexpected genetic modification, which is called the off-target effect.57 The CRISPR/Cas-9 target efficiency is determined by the 20-nucleotide sequences of sgRNA and the PAM sequences adjacent to the target genome. It has been shown that more than three mismatches between the target sequence and the 20-nucleotide sgRNA can result in off-target effects.64 The off-target effect can possibly cause harmful events such as sequence mutation, deletion, rearrangement, immune response, and oncogene activation, which limits the application of the CRISPR/Cas-9 editing system for therapeutic purposes.65 To mitigate the possibility of CRISPR/Cas-9 off-target effect, several strategies have been developed, such as optimization of sgRNA, modification of Cas-9 nuclease, utilization of other Cas-variants, and the use of anti-CRISPR proteins.66 Selecting and designing an appropriate sgRNA for the targeted DNA sequence is an important first step to reduce the off-target effect.67 When designing sgRNA, strategies such as GC content, sgRNA length, and chemical modifications of sgRNA must be considered. Generally speaking, studies revealed that GC content of between 40% and 60%, truncated (short length of sgRNA), and incorporation of 2-O-methyl-3-phosphonoacetate in the sgRNA ribose-phosphate backbone are the preferred methods to increase genome editing efficiency of CRISPR/Cas-9.67,68 Modifying the Cas-9 protein to optimize its nuclease specificity is another way to reduce off-target effects. For instance, mutating either one of the catalytic residues of Cas-9 nuclease (HNH and RuvC) will convert the Cas-9 into nickase that could only generate a single-stranded break instead of a blunt cleavage.69 It has been reported that the use of the inactivated RuvC domain of Cas-9 with sgRNA can reduce the off-target effect by 100 to 1500 times.70 Moreover, the nuclease Cas-12a (previously known as Cpf1) is a type V CRISPR/Cas system that provides high genome editing efficiency.71 Unlike the CRISPR/Cas-9 system, CRISPR/Cas-12a can process pre-crRNA into mature crRNA without tracrRNA, thereby reducing the size of plasmid constructs. The Cas-12a protein recognizes a T-rich (5-TTTN) PAM sequence instead of 5-NGG and provides high accuracy at the target gene loci than Cas-9.69 Recently, the use of multicomponent Class I CRISPR proteins, such as CRISPR/Cas-3 and CRISPR/Cas-10 provides better genome editing efficiency than Cas-9.72 The Cas-3 is an ATP-dependent nuclease/helicase that can delete a large part of DNA from the target site without prominent off-target effect. For instance, the DMD gene were repaired by Cas-3-mediated system in induced pluripotent stem cell.73 The Cas-10 protein does not require the PAM sequence and can identify sequences even in the presence of point mutation.72 Anti-CRISPR (Acr) proteins are phage derived small proteins that inhibit the activity of CRISPR/Cas system. They are a recently discovered method to reduce off-target effects of CRISPR/Cas-9.74 From Acr proteins, AcrIIA4 specifically targets Cas-9 nuclease. AcrIIA4 mimics DNA and binds to the Cas-9 site, making impossible to perform further cleavage in area outside the target region.75 Furthermore, CRISPR/Cas-9 gene editing has been challenged by ethics and safety all over the world. Since the technology is still in its infancy and knowledge about the genome is limited, many scientists restrain that it still needs a lot of work to increase its accuracy and make sure that changes made in one part of the genome do not have unforeseen consequences, especially in the application towards human trials.52

AAVs, adeno-associated viral vectors; ABE, adenine base editor; Acr, anti-CRSPR; AVs, adeno-viral vectors; ATP, adenosine tri-phosphate; BCL11A, B-cell lymphoma 11 A; CAS-9, CRISPR-associated protein-9; CBE, cytidine base editor; CCR5, chemokine receptor type 5; CFTR, cystic fibrosis conductance transmembrane receptor; CRISPR, clustered regularly interspaced short palindromic repeat; CrRNA, CRISPR ribonucleic acid; DMD, Duchenne muscular dystrophy; DNA, deoxyribonucleic acid; DSBs, double-stranded breaks; HDR, homology-directed repair; LVs, lentivirus vectors; NHEJ, non-homologous end Jjining; PAM, protospacer adjacent motif; PD-1, programmed cell death-1; RNA, ribonucleic acid; TALENs, transcriptionactivator like effector nucleases; TRAC, T-cell receptor alpha; TRBC, T-cell receptor beta; TracrRNA, trans-activating CRISPR ribonucleic acid; ZFNs, zinc finger nucleases.

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Mechanism and Applications of CRISPR/Cas-9-Mediated Genome Editing

Recommendation and review posted by Bethany Smith

Life Extension is a company that sells supplements and lab tests through its website. Customers can contact the customer support team at any time, but accessing a products certificate of analysis can be difficult.

Life Extension has been in business for more than 40 years. It claims that it sells science-backed products, such as vitamins, moisturizers, and functional foods that help target different health concerns, including weight gain, signs of skin aging, and digestive issues.

Regular customers can opt for the Autoship & Save program, which allows them to receive products every 30 or 60 days.

The company has a 12-month return policy. Customers who are unsatisfied with their purchase can return their item within this time frame. However, many customers claimed they found returning unwanted products very difficult.

Life Extension has a 1 out of 5 rating on Better Business Bureau (BBB) and 2.5 stars on Trustpilot. Customers wrote that they received a wrong order and some products caused frequent headaches, but they could not return them.

The company website does not provide lab test documents.

Positive reviews state that products are affordable and do not have a lot of fillers.

In 2017, the Food and Drug Administration (FDA) issued a warning letter stating that Life Extension stocks products as medications, which goes against its regulations. Life Extension suggests buying astragalus, blueberry, vitamin D, and other products from the website to treat breast cancer.

It also shared a post that mentions other items that prevent heart failure.

Life Extension may suit people looking to buy supplements that meet their daily recommended intake.

The website may serve as a one-stop shop, as it stocks vitamins, home tests, and functional foods. Individuals may use the service if they are buying different product formulations without checking multiple companies.

Vegetarians may find products that do not contain animal ingredients. However, these may still include beeswax or eggs.

Life Extension has pros and cons:

These are some of the products that a person can find on Life Extensions website:

The Female Basic Hormone Panel Blood Test measures the levels of different hormones, including estradiol, progesterone, and dehydroepiandrosterone sulfate (DHEA-S).

People do not have to fast to use this test. Life Extension recommends that people who supplement with hormones, including hormone replacement therapy (HRT), take their medication 2 hours before doing the test. The company also suggests doing the blood test between 8.0010.00 a.m.

In addition, Life Extension recommends individuals who are premenopausal take a blood sample on day 21 of their menstrual cycle. However, those who are postmenopausal can do the test on any day of the month.

This thyroid lab test provides an overview of thyroid function, as it measures the levels of:

There is no need to fast to use this test. Persons with low thyroid function may have drier skin, feel tired, and experience constipation.

Life Extension stocks a wide range of energy supplements. These are two products that people can buy to fight fatigue and feel energized.

These capsules contain is nicotinamide riboside, which may help increase energy levels and support heart health.

It also has resveratrol, which according to Life Extension, helps maintain longevity. A 2021 study notes that this may act as a life-extending agent, as it increased adult longevity in flies. However, more studies are needed.

A 2020 literature review mentions the side effects that resveratrol may cause:

The supplements are vegetarian, gluten, and GMO-free.

These capsules contain vitamin B6, sodium, carnosine, and pyrroloquinoline quinone (PQQ). PQQ is a vitamin that may aid longevity. It is naturally present in fruits and vegetables, such as green peppers, kiwi, and spinach.

A 2021 study found that PQQ has vitamin, anti-inflammatory, and antioxidant qualities that may promote well-being, control symptoms, and prevent some illnesses.

The product is GMO and gluten-free.

Life Extension offers digestive supplements for people who may have gastric discomfort. It states that these products may help aid digestion, break down macronutrients, and relieve discomfort after eating a large meal.

A person can buy softgels, capsules, or vegetarian capsules.

These are two of the top digestive supplements:

Bloat Relief softgels are for individuals who regularly experience bloating, uncomfortable digestion, or occasional gas.

Ingredients include ginger extract, fennel seed oil, and turmeric.

The researchers behind a 2019 study note that ginger has anti-inflammatory, antioxidant, and antitumor characteristics.

A 2020 review indicates that ginger may help relieve nausea and improve morning sickness in persons undergoing cancer treatment.

The National Center for Complementary and Integrative Health states that large doses of ginger can cause some side effects:

Life Extension recommends these supplements for people who follow a plant-based diet. They contain vegetarian-friendly enzymes, including lactase, lipase, and xylanase.

Lactase helps the body digest lactose, a type of sugar that is present in dairy products.

They are vegetarian and GMO-free.

Life Extension stocks products that moisturize and restore the skin by reducing the appearance of wrinkles and fine lines and increasing hydration.

These are the top skin care products available on the website:

This serum supports healthy aging and contains many ingredients, including ceramides, green tea leaf extract, and sunflower seed oil.

Ceramides have water-retaining properties that help hydrate the skin. Hydration can help reduce the appearance of fine lines that may be more visible on dry skin.

Some companies also stock sunscreens containing ceramides to help protect the skin from UV.

The serum is paraben-free and not tested on animals.

This stem cell cream contains many ingredients, including lactic acid, citric acid, lecithin, and seaweed extract. It helps protect the skin from environmental irritants.

The FDA states that lactic acid helps cleans the pores, reduce dark spots, and improve skin texture.

Life Extension provides other services:

Individuals who are interested in using Life Extensions services may consider the following:

People who wish to start taking vitamins may consider consulting a doctor to discuss their safety, as some supplements interact with other medications.

They may also see a doctor if they have symptoms of digestive problems, including:

These are commonly asked questions about Life Extension.

Life Extension has a low rating on BBB and Trustpilot. Reviews state that there are no lab tests on the company website, and returning a product may be problematic.

The company has also received a warning letter from the FDA, as it sometimes claims that its products can cure or treat health conditions.

Life Extension states that its vitamins are of high quality, and it only uses raw materials. However, products do not come with a lab test result. The company asks customers to call the customer service team to provide them with the document.

It is from the United States but gets raw materials from Japan and Europe.

According to the brand, Life Extension produces its vitamins in the U.S. in line with good manufacturing practices.

Life Extension is well-known for its supplements. It sells supplements that may help people experiencing stomach discomfort, bloating, and fatigue. It also has a range of skin care products that support healthy aging.

Lab tests, such as thyroid and hormone tests, are available to order on its website.

A person may have to consult their doctor first before buying any supplements. These may not be safe, especially for those who take other medications.

Originally posted here:
Life Extension Review 2023 - Medical News Today

Recommendation and review posted by Bethany Smith

Most people want to live a long and happy life or at least avoid a short and miserable one. If youre in that majority, then youre in luck. Over the last decade, a quiet research revolution has occurred in our understanding of the biology of ageing.

The challenge is to turn this knowledge into advice and treatments we can benefit from. Here we bust the myth that lengthening healthy life expectancy is science fiction, and show that it is instead scientific fact.

Theres plenty of evidence for the benefits of doing the boring stuff, such as eating right. A study of large groups of ordinary people show that keeping the weight off, not smoking, restricting alcohol to moderate amounts and eating at least five servings of fruit and vegetable a day can increase your life expectancy by seven to 14 years compared with someone who smokes, drinks too much and is overweight.

Cutting down calories even more - by about a third, so-called dietary restriction - improves health and extends life in mice and monkeys, as long as they eat the right stuff, though thats a tough ask for people constantly exposed to food temptation. The less extreme versions of time-restricted or intermittent fasting only eating during an eight-hour window each day, or fasting for two days every week is thought to reduce the risk of middle-aged people getting age-related diseases.

You cant outrun a bad diet, but that doesnt mean that exercise does not do good things. Globally, inactivity directly causes roughly 10% of all premature deaths from chronic diseases, such as coronary heart disease, type 2 diabetes and various cancers. If everyone on Earth got enough exercise tomorrow, the effect would probably be to increase healthy human life expectancy by almost a year.

But how much exercise is optimal? Very high levels are actually bad for you, not simply in terms of torn muscles or sprained ligaments. It can suppress the immune system and increase the risk of upper respiratory illness. Just over 30 minutes a day of moderate to vigorous physical activity is enough for most people. Not only does that make you stronger and fitter, it has been shown to reduce harmful inflammation and even improve mood.

However fit you are and well you eat, your immune system will, unfortunately, get less effective as you get older. Poor responses to vaccination and an inability to fight infection are consequences of this immunosenescence. It all starts to go downhill in early adulthood when the thymus a bowtie-shaped organ in your throat starts to wither.

That sounds bad, but its even more alarming when you realise that the thymus is where immune agents called T cells learn to fight infections. Closing such a major education centre for T cells means that they cant learn to recognise new infections or fight off cancer effectively in older people.

You can help a bit by making sure you have enough key vitamins, especially A and D. A promising area of research is looking at signals that the body sends to help make more immune cells, particularly a molecule called IL-7. We may soon be able to produce drugs that contain this molecule, potentially boosting the immune system in older people. Another approach is to use the food supplement spermidine to trigger immune cells to clear out their internal garbage, such as damaged proteins, which improves the elderly immune system so much that its now being tested as a way of getting better responses to COVID vaccines in older people.

Senescence is a toxic state that cells enter into as we get older, wreaking havoc across the body and generating chronic low-grade inflammation and disease essentially causing biological ageing. In 2009, scientists showed that middle-aged mice lived longer and stayed healthier if they were given small amounts of a drug called rapamycin, which inhibits a key protein called mTOR that helps regulate cells response to nutrients, stress, hormones and damage.

In the lab, drugs like rapamycin (called mTOR inhibitors) make senescent (aged) human cells look and behave like their younger selves. Though its too early to prescribe these drugs for general use, a new clinical trial has just been set up to test whether low-dose rapamycin can really slow down ageing in people.

Discovered in the soil of Easter Island, Chile, rapamycin carries with it significant mystique and [has been hailed] in the popular press as a possible elixir of youth. It can even improve the memory of mice with dementia-like disease.

But all drugs come with pros and cons and as too much rapamycin suppresses the immune system, many doctors are averse to even consider it to stave off age-related diseases. However, the dose is critical and newer drugs such as RTB101 that work in a similar way to rapamycin support the immune system in older people, and can even reduce COVID infection rates and severity.

Completely getting rid of senescent cells is another promising way forward. A growing number of lab studies in mice using drugs to kill senescent cells - so-called senolytics - show overall improvements in health, and as the mice arent dying of disease, they end up living longer too.

Removing senescent cells also helps people. In a small clinical trial, people with severe lung fibrosis reported better overall function, including how far and fast they could walk, after they had been treated with senolytic drugs. But this is only the tip of the iceberg. Diabetes and obesity, as well as infection with some bacteria and viruses, can lead to more senescent cells forming. Senescent cells also make the lungs more susceptible to COVID infection, and COVID makes more cells become senescent. Importantly, getting rid of senescent cells in old mice helps them to survive COVID infection.

Ageing and infection are a two-way street. Older people get more infectious diseases as their immune systems start to run out of steam, while infection drives faster ageing through senescence. Since ageing and senescence are inextricably linked with both chronic and infectious diseases in older people, treating senescence is likely to improve health across the board.

It is exciting that some of these new treatments are already looking good in clinical trials and may be available to us all soon.

More here:
Life extension: the five most promising methods so far

Recommendation and review posted by Bethany Smith

Do you want to live a better, healthier and longer life? Me too.

Lets go back to 1937, when Albert Szent-Gyrgyi won a Nobel Prize for his discovery of ascorbic acidvitamin Cthat enables the body to efficiently use carbohydrates, fats, and protein (I use it a lot during cold and flu season, you?). It was a massively consequential discovery, as it not only saved and extended countless lives, but it also contributed to the foundations of modern nutrition. Szent-Gyrgyi, himself, was blessed with a long life; he died in 1986 at the age of 93. But he might just as well be known for what he said on his 90th birthday: I wish I could be 75 again!

No doubt, that comment elicits more than a few eyerolls today. Especially since the CDC has recently downgraded American life expectancy to just 77 years. But could 75 someday be the new 40an age by which, like Szent-Gyrgyi, were only hitting our stride? Well, if the burgeoning activity of the life extension industry is any indication, we may actually be on the cusp of making it soand enjoying life to the fullest right up to the extended end. Which brings us to the morbid thought of mortalitythat end state most of us seek to delay, if not dodge.

It may strike many as common sense that most causes of death are what we have come to understand as age-related: The longer we live, the more likely we are to develop, for example, heart disease, cancer, or Alzheimers. Therefore, a reasonable thing one can do to prevent the development of age-related diseases, is to, well, not age. It turns out thats actually not as flippant as it sounds. So, is that possible and how do we get there?

In short, all strategies for life extension revolve around a combination of three factors, all working together to fortify health and wellness for a longer haul: 1) things you should stop doing (I have a list which I mostly ignore, you?), 2) things you should start doing (honestly part of my same list), and 3) adopting the contributions of new health and wellness technologies and scientific discoveries that are helping to curb aging. All three comprise parts of a budding ecosystem that is growing into a multibillion-dollar industry on an exponential trajectory to displace everything we have come to understand as modern medicine. If just what we know today were fully embraced and actualized, the global economy could also be transformedand with it, a renaissance of human flourishing.

And we could use it. When ranking countries for life expectancy, the United States often doesnt make the top 50despite having the highest healthcare costs per capita in the world, by far. In other words, maybe were doing it wrong.

Clearly, somethings got to give. Right?

None of this is lost on the investment community, which is set to plow billions into a nascent industry that is fast approaching its inflection point. Driving that inflection is a fundamental pivot from treating symptomsthe bread and butter of the rapidly collapsing medical-industrial complexto addressing the root causes of aging and disease. Its a shift that is ushering in a new and immensely disruptive paradigm that some analysts envision creating a global market approaching $300B by 2030. Its likely bigger when you add in personalized skin health and beautynot to mention food.

Again, like many such watershed moments, this is a story of convergences. Its a perfect storm comprising new and surprising discoveries about the workings of the human body, myriad innovative technologies, coupled with an ever-increasing geriatric population. Now add to this mix an explosion in chronic diseases and a growing demand for more Personalized, Precise, Preventive, and Participatory aging therapeutics (the so-called 4 Ps) and we have a movement.

Beyond its potential for profitability, though, the longevity movement benefits from a certain intrinsic appeal: it could be a tremendous positive impact for humanity. Startups and leading global organizations are diligently at work developing the next level of health, beauty and wellness products, services and treatmentsfrom delivering mass-personalization AI technology to help consumers choose at Revieve, to science-driven wellness at Thorne HealthTech, to curing Alzheimers at Genentechall with investments beginning to show promise.

Another leading catalyst in shifting the world away from the traditional disease-focused regime in favor of targeting root causes of age-advancing chronic illnesses, is Viomea life sciences company that has pioneered the field of human gut microbiome diagnostics and treatment. I took a look under the hood to learn whats behind this longevity science and how are they rebooting an industry.

The big idea behind Viomes science is simultaneously cutting edge and ancient (stay with me here as we unpack this). It was actually first articulated in the 4th century BCby Hippocrates of Kos. Let food be thy medicine, he proclaimed, and medicine be thy food. As investors in next-gen food companies, like Manna Tree, will tell youhe was onto something.

You can think of the gut microbiome as the Chief Architect of your health. The trillions of microbes that reside in your gut help you digest food, absorb nutrients, maintain a healthy weight, neutralize toxins, fight off bad bacteria, and many other functions that are responsible for keeping you alive and healthy. How the microbes in your gut respond to the food you eat creates a chain reaction that can be beneficial to you or, if you're eating the wrong foods, can promote inflammatory activity and microbial imbalance, or, dysbiosis. In other words, the very conditions that work to curtail lifespan.

It turns out that illnesses such as cancer, diabetes, heart disease, stroke, Parkinsons, Crohns, Alzheimers, arthritis, depression, and many other debilitating conditions share a common denominatorschronic inflammation for one. In fact, chronic inflammation is both a precursor and prerequisite to the onset and development of todays deadliest diseasesand aging. And all these conditions are pointing to the gut.

Its to these ends that Viomes 50+ health scores reveal ones underlying contributors to poor healthand more importantly, to identifying the microbiome-based correctives that can extend both lifespan and healthspan. Affirming both Hippocrates and Viome, Dr. Sherry A. Rogers astutely quipped, The road to health is paved with good intestines!

In a free-market, innovation-economy, Viome is certainly not alone in this endeavor. Life Biosciences and many others are also targeting the biology of aging. The emerging sector operates on the premise that aging is in fact modifiable, being caused by biological mechanisms that can be targeted therapeutically. Elysium is another. It is working to make practical the myriad scientific advancements in aging research that in turn are leading to compelling new developments that will, fundamentally redefine every aspect of how we think about healthcare. Sampo Parkkinen, Revieve's Founder and CEO agrees and believes that the world's leading brands and retailers can help consumers personalize their health, beauty, and wellness through relationship-driven commerce: "Bringing personalized solutions to worlds consumers must go beyond healthcare, into the daily products, services, and treatments people use, from skincare and beauty products to supplements and food, it should all be on the table where consumers shop, he comments."

The key to moving life expectancy forward is turning back the biological clock. The old clich goes, If I knew I was going to live this long, Id have taken better care of myself! Or, as one of my Moms favorites, actress Mae West, said, Youre never too old to become younger. The good news is that theres no time like the present to reboot and begin preparing for a healthy future. And lifestylemarked by a nutritionally-rich diet (most tell us to ditch the fast food, but not everyone can), proper exercise (yes, there is an inverse relationship between physical activity and mortality), adequate sleep (I dont get enough, you?), good stress management (how are you doing?)is also central to any life extension regime. We have been told all of this before, yet we still struggle.

That said, a central goal of lifestyle modification is not only to reduce the incidence of disease but promote healthy, successful aging. Its true that ones biological age need not match chronological age. Consider Viome founder Naveen Jain. I am 63 years old, he says, but my biological age is now 10 years younger. I have achieved this in the last two years, simply by understanding how my body works, using Viomes tools, which tell me what to eat and why and what to avoid and why. It then details the specific nutrients my body needs to live a disease-free life.

In addition to its AI-driven gut microbiome tests, the company is beginning to produce personalized supplements and formulas that combine the necessary nutrients into daily capsules, thus eliminating the need to stock dozens of products. Consequently, Naveen actually does anticipate reaching 75 but registering a biological age of just 40. Not that he expects to age in reverse, a la The Curious Case of Benjamin Button. Nor is this about achieving immortality. Rather, longevity science is really more about enabling people to be as healthy as possible for as long as they are alive. It's about having the strength, energy, and vitality to do what they want to do, even possibly at 100.

I know what youre thinking, Naveen is the CEO and didnt actor George Burns live to be 100 while famously smoking cigars every day? Yes, however, Ive watched his positive transformation over the years, along with Viome customer data, the results are remarkably noticeable (but alas, Im not a healthcare professional). The early industry data now has supplement manufacturers, and companies like Amway, taking note and working determine how to globally scale and deliver personalized nutrition through mass-customization. As I have said at Northwestern Universitys Kellogg School of Management, look for a variety of co-created innovation and delivery partnershipsbetween upstarts and global 1000sin this space starting next year.

Want another view on potential impacts? Longer lifespans are an effect of the measures anyone can take to improve their healthand it seems to also be an emerging societal and economic imperative. Consider, for example, if everyone in the world were to lower their biological age by even a few years, the global disease burden would be reduced dramatically. Today, according to the CDC, six in 10 U.S. adults suffer chronic health conditions; four in 10 have two or more of these diseases. Some of these diseases are both debilitating and expensive. According to estimates by the World Economic Forum, between 2010 and 2030, the total cost to the worlds health systems will approach $50T (yes trillions). Could we cut that in half? The leaders in the life extension spaceboth startups and the big-dogsbelieve so. And supporting data is mounting.

Looking for a bottom line? Me too, hows this: if you can maintain good gut health, then that good gut health will keep the rest of you healthy. If, on the other hand, the gut microbiota are in dysbiosis, then youre up for diseases caused by the chronic inflammation that inevitably resultsthe very conditions that are know to slash human life expectancy by half.

When the experts declare the upper limits of life expectancy, we believe it like some law of the universe. When someone dies at, say, 80, we accept that theyve lived a good long life. Weve become conditioned to believe it, and like so many other things connected to ones mindset, it becomes a self-fulfilling prophecy. Normal cholesterol in a society where its normal to drop dead of a heart attack really should not be considered a good thing. Right? We dont achieve more than we expect. But if we do expect more, it must begin with an understanding of whats really going on in the body. We are starting to do that now, thanks to the biological, cellular, and genetic ground truths established by the sciences and technologies advanced by the likes of Viome, Life BioSciences, Elysium, and many other emerging players the world will soon hear about.

But what about genes? you might ask. Surely, ones genetic makeup affects lifespan, right? Interestingly, we are learning that genetics may be responsible for only a very small percentage of human diseasesthe vast majority are triggered by environmental and lifestyle factors. And both are of tremendous consequence to the makeup of ones gut microbiome, which, we are now understanding, can be the chief driver of genetic expression (or epigenetics), turning genes on and off depending upon which microbes are present in the gut.

So, while some people do hit the genetic lottery, like George Burns, the rest of us can improve our luck through lifestyle choices. But the differencenow those choices need to be informed. This is where Viomes Human Gene Expression test comes in. With its origins in biodefense science developed at Los Alamos National Lab, it reveals a true state of ones overall health, the measurable potential for developing disease, and the nutritional corrective actions one can take to restore the homeostasis that is essential to healthy longevity. Again, it seems if you can improve the health of your gut microbiome, you could also take some of your genetic fate into your own hands.

In the end, though, when all is said and done, given the events of the past few years, many have learned that there is more to life than increasing its length. To this end, we would all do well to heed the words of the 14th century holy man who wrote, It is vanity to wish for length of life, and to care little that the life should be well spent. Indeed, if were to be older, well also need to be wiser.

More:
Want To Live Longer? How Life Extension Industry Will Reboot ... - Forbes

Recommendation and review posted by Bethany Smith

cryonics, the practice of freezing an individual who has died, with the object of reviving the individual sometime in the future. The word cryonics is derived from the Greek kros, meaning icy cold.

Cryonic preservation can be performed only after an individual has been declared legally dead. The process is initiated shortly after death, the body being packed in ice and shipped to a cryonics facility. There the blood is drained from the body and is replaced with antifreeze and organ-preserving compounds known as cryoprotective agents. In this vitrified state, the body is placed in a chamber filled with liquid nitrogen, where it will theoretically stay preserved at -196 C until scientists are able to find a way to resuscitate the body in the future.

A chamber filled with liquid nitrogen in which individuals are preserved at the Alcor Life Extension Foundation in Scottsdale, Arizona.(more)

Cryonic preservation is expensive, full-body preservation potentially costing hundreds of thousands of dollars. Nonetheless, by 2023 about 500 individuals had been cryonically preserved, the majority of them in the United States. Dozens of pets had also been preserved. Some individuals chose to have their entire bodies frozen, whereas others wanted only their heads preserved, a process known as neuropreservation. The option to cryonically preserve only a persons head is based on the belief by many cryonics adherents that cryonically preserved personalities may one day be downloaded into robot bodies or be transferred into entirely new bodies grown from stem cells.

The concept of cryonic preservation was popularized in The Prospect of Immortality, a book by Robert Ettinger that was initially released in 1962 and formally published in 1964. Ettinger subsequently became known as the father of cryonics. His body was cryonically preserved upon his death in 2011 and was stored at the Cryonics Institute in Clinton Township, Michigan. The first human to be cryonically preserved was James Bedford. On January 12, 1967, Bedford died from liver cancer that had metastasized to his lungs. Bedford died before all the arrangements for his cryonic preservation could be completed. As a result, his body was injected with cryoprotective agents without first draining his blood, and his body was then packed in dry ice. Bedfords body was later immersed in liquid nitrogen and transferred from one facility to another, finally ending up at the Alcor Life Extension Foundation in Scottsdale, Arizona.

The rest is here:
Cryonics | Description, Process, Popularization, & Facts

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It is most important that the cryonics patient is a CI member and has completed all of the necessary documents with their funding arrangements in place. This will allow CI to accept the patient without hesitation or delay. Time is of the essence for a successful cryopreservation and being prepared before an emergency arises can make all the difference. If a patient is near death, the Cryonics Institute should be notified immediately at 1-586-791-5961 and kept updated of any changes.

It is also suggested that, if possible, a cryonics patient relocate themselves to Michigan, near the CI facility, before death occurs. Delay in transfer to the facility can be avoided this way. If this is not an option, it is strongly advised that the CI member, or their next of kin, make arrangements with a local funeral director before the time of need. The funeral directorcan then be better prepared to act during an emergency.

Initial Cool-down and Transport.

If possible, CI Members should arrange to have a cryonics standby team standing by their bedside when they are in a terminal condition. Such a team can initiate rapid cool-down and provide other helpful stand by procedures.

The patient should be pronounced dead as soon as possible after clinical death (which usually means after cessation of heartbeat and breathing). The patient should then be cooled immediatelyespecially the headby application of ice. A slurry of ice water can cool much faster than ice cubes alone, so an inflatable basin for giving shampoos can be filled with ice and water to cool the head. Even better would be cooling the entire body of the patient in a body bag filled with ice water. The best scenario is for the patient to die at home under hospice care, with trained personnel morticians or Cryonics Institute (CI) volunteers on hand. (No guarantee is made that CI volunteers can be found.)

An anticoagulant should be injected to prevent the blood from clotting, which will help to improve the patients perfusion. When using the anticoagulant, Heparin, a dose of 30,000 units is given for patients weighing up to 200 lbs. and a dose of 40,000 units is given for patients weighing 200 lbs. or more. Once the heparin is injected, Cardio Pulmonary Support (CPS) (chest compressions) is required for at least 5 minutes to circulate the heparin throughout the body.

During transportation from the place of death to the funeral home if the patient dies outside of Michigan, or to the CI facility if the patient dies in Michigan, CPS should be given, if feasible, manually or by machine (thumper or Lucas) in order to minimize deterioration, help cool the patient, and to help distribute the heparin.

If the patient can be moved to Michigan near the CI facility before death occurs, this will eliminate transportation delays. Patients living outside of Michigan will require the services of a funeral director near them to cool the patient in ice and arrange for transportation to Michigan. The out of state funeral director will obtain the necessary transit permits and arrange for the patient to be transported to Michigan, by airline or vehicle, whichever is fastest, while keeping the patient in ice. If the patient is flown to Michigan, CI personnel will arrange to have the patient picked up from the Detroit Metro Airport (DTW) and brought back to the CI facility.

Perfusion

Upon arrival at CIs facility, the patients blood is removed as CryoProtectant Agents (CPAs, substances that prevent ice formation) are introduced to replace the patients blood and body water, a process known as Perfusion. Blood vessels are accessed and cannulated for the perfusion and a licensed funeral director performs the perfusion.

The cryoprotectant used by CI is called CI-VM-1, a Vitrification Mixture which was developed by CIs former in-house cryobiologist, Dr.Yuri Pichugin. Vitrification solutions can completely eliminate ice formation. The perfusion with vitrification solution is done at increasing concentrations, until a target concentration of 70% CI-VM-1 is reached. The patient is kept cold through the process, as the lower concentration CPAs are stored and introduced at refrigerator temperature. The 70% CI-VM-1 has been stored in a freezer, so it is below freezing temperature. Thermocouples are placed in the nasopharynx for monitoring the patients brain core temperature through the perfusion.

CI protocol is to perfuse the entire body, by way of the carotid arteries. The funeral director gains access to the blood vessels by small incisions along the clavicle. The right and left carotid arteries are carefully incised for the insertion of the cannulas required for perfusion. Both arteries are cannulated towards the heart and towards the brain to achieve a full body perfusion. To protect the integrity of the vascular system and ensure a successful perfusion, the pressure and speed at which the cryoprotectant is introduced into the patient is monitored very carefully.

The right and left jugular veins are also cannulated for drainage and for sampling of the effluent. A refractometer is used to measure the refractive index of the effluent from the jugular veins. Perfusion continues until the refractive index of the effluents is matched with the refractive index of CI-VM-1. Once this is achieved, the perfusion is halted. Perfusion will be halted before the refractive index of CI-VM-1 is reached only if there is considerable edema evident in the patients brain.

When the perfusion is complete, the incisions are sutured by the funeral director. The patient is then sheathed with a light weight cover for dignity and placed in an insulation pouch before being moved from the operating table to a stretcher where they are secured to a backboard for support. The stretcher is used to transport the patient from the perfusion room to the computer controlled cooling unit, inside the CI facility.

Further Cool-Down and Storage

The patient is carefully placed in the cooling unit on their backboard. The insulation pouch is opened slightly to allow for consistent cooling. The appropriate program is selected to steadily cool the patient to liquid nitrogen temperature. The process of cooling the patient to -196c takes five and a half days. The cooling is done by CIs computer-controlled cooling unit. The computer-controlled cooling unit constantly monitors the temperature inside the cooling unit via a thermocouple.

At the end of the cooling process, the patient is carefully removed from the cooling unit and the insulation pouch is closed. Once ID tags are attached to the patient for identification purposes, the patient is ready to be transferred to the cryostat (long term storage unit). The patient transfer is done quickly, but carefully, while the insulation pouch is saturated with nitrogen, so there is no appreciable warm-up during transfer. The ropes that are attached to the patients backboard are secured to an electric lift and the patient is safely lowered into the cryostat. Cryostat liquid nitrogen levels are monitored daily to ensure the safety of the patients. CI staff adds liquid nitrogen from the facilitys bulk tank to the cryostats when needed.

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Guide to Cryonics Procedures - The Cryonics Institute

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The adrenal glands are part of what is called the HPA (hypothalamic-pituitary-adrenal) axis, which controls how your body reacts to stress. When faced with a stressful situation, your adrenal glands release a cascade of hormones, including adrenaline, DHEA, and cortisol as part of your fight-or-flight response. Once the threat has passed, your body processes normalizeyour heartbeat and breathing slow to their usual rate, your muscles relax, and your adrenals turn off production of those hormones until they are needed again.

In cases where stress becomes chronic, there can be a seemingly constant flow of stress hormones, that in turn, overwhelm your body and contribute to brain health/mental health issues. Chronically high cortisol levels also cause your blood sugar and insulin levels to spike. This can lead to harmful changes in the brain, including a drop in the calming neurotransmitter serotonin. High levels of cortisol are linked to mental health conditions, such as anxiety, depression, and PTSD.

In the brain, chronic stress produces more white matter and fewer neurons (gray matter) than normal, skewing their balance and disrupting communication within the brain.

More:
Hormone Evaluation and Replacement Therapy | Amen Clinics

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BackWhat Is It, Causes, Signs and Symptoms, and More

Author: Anna Hernndez, MD

Editors: Alyssa Haag, Emily Miao, PharmD, Kelsey LaFayette, DNP, RN

Illustrator: Jessica Reynolds, MS

Copyeditor: Sadia Zaman, MBBS, BSc

Hypogonadism is a clinical syndrome that occurs when the gonadstestes and ovariesproduce low levels of sex hormones due to a disruption of the hypothalamic-pituitary-gonadal (HPG) axis.

There are two main types of hypogonadism, primary and secondary. Primary hypogonadism is caused by dysfunction of the gonads, and can be acquired or congenital. Acquired causes include radiation therapy, chemotherapy, autoimmunity, trauma to the gonads, and certain infections, like mumps orchitis. On the other hand, congenital causes include genetic disorders, like Klinefelter syndrome or Turner syndrome, both of which affect gonadal function. Regardless of the cause, the result is a decrease or complete absence of sex hormones, which means there is no negative feedback on the hypothalamic-pituitary-gonadal axis. This leads to an overproduction of the LH and FSH gonadotropins, thus giving primary hypogonadism its other name, hypergonadotropic hypogonadism.

Signs and symptoms of hypogonadism vary depending on whether hypogonadism occurs before or after puberty. The most common presenting feature of hypogonadism in teenagers is a delay in puberty, which generally occurs when puberty has not started by age 13 in those assigned female at birth and age 14 in those assigned male at birth. Other clinical features of hypogonadism include a high-pitched voice, sparse body hair, poorly developed muscles, small or underdeveloped genitals, and short stature due to delayed epiphyseal closure. Another presenting feature can be primary amenorrhea, which is when an individual hasnt had their first menstruation by the age of 13 to 15.

Diagnosis of hypogonadism begins with a thorough medical history and physical exam, including assessment using the Tanner scale. The Tanner scale, or Tanner stages, consists of a predictable set of steps that individuals go through as they develop primary and secondary sex characteristics and become sexually mature. This scale centers on two, independent criteria: the appearance of pubic hair; and the increase in testicular volume and penile size and length in those assigned male at birth or with breast development in those assigned female at birth.

Treatment of hypogonadism is directed at addressing the underlying cause, when possible. In most cases, hypogonadism can be managed with hormone replacement therapy to ensure the onset and progression of puberty, as well as improve the symptoms of hypogonadism.

Certain cases of hypogonadism are transient and can be reversed, especially if they are a result of a treatable underlying disorder, such as malnutrition, excessive exercise, or stress. In addition, there have been cases of idiopathic hypogonadotropic hypogonadism that have reversed over time, although the exact reason why is still unknown.

Hypogonadism refers to a clinical syndrome that results in low levels of sex hormones. There are two types of hypogonadism: primary and secondary. Signs and symptoms of hypogonadism depend on the time of onset, and can include delayed puberty, primary amenorrhea, changes in mood and energy, infertility, decreased libido, and erectile dysfunction. Diagnosis of hypogonadism is based on the clinical manifestations along with lab tests to assess the levels of sex hormones in the body. Treatment involves long-term hormone replacement therapy and fertility treatments, as well as treatment of the underlying cause of hypogonadism, when possible.

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Hypogonadism: What Is It, Causes, Signs and Symptoms, and More - Osmosis

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Miguel ngel Perales, oncologist: What we are doing today with CAR-T cell therapy against cancer seems like science fiction  EL PAS USA

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Miguel ngel Perales, oncologist: What we are doing today with CAR-T cell therapy against cancer seems like science fiction - EL PAS USA

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Depression can hit hard during the winter months, here's how to prevent it  NewsCenterMaine.com WCSH-WLBZ

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Depression can hit hard during the winter months, here's how to prevent it - NewsCenterMaine.com WCSH-WLBZ

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As a study finds a stable weight is key to living longer, a leading nutritionist's guide on how to... Escape t  Daily Mail

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As a study finds a stable weight is key to living longer, a leading nutritionist's guide on how to... Escape t - Daily Mail

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Fortress Biotech Subsidiary Helocyte Announces Option Agreement with City of Hope for Exclusive Worldwide Rights to use a Novel Bispecific CMV/HIV CAR T Cell Therapy (optionally in combination with Triplex) for the Treatment of Adults Living with HIV-1  Yahoo Finance

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Fortress Biotech Subsidiary Helocyte Announces Option Agreement with City of Hope for Exclusive Worldwide Rights to use a Novel Bispecific CMV/HIV CAR...

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Youve probably read stories about new research using the gene editing technique CRISPR, also called CRISPR/Cas9. The scientific world is captivated by this revolutionary technology, since it is easier, cheaper and more efficient than previous strategies for modifying DNA.

The term CRISPR/Cas9 stands for Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR associated protein 9. The names reflect important features identified during its discovery, but dont tell us much about how it works, as they were coined before anyone understood what it was.

CRISPR/Cas9 is a system found in bacteria and involved in immune defence. Bacteria use CRISPR/Cas9 to cut up the DNA of invading bacterial viruses that might otherwise kill them.

Today weve adapted this molecular machinery for an entirely different purpose to change any chosen letter(s) in an organisms DNA code.

We might want to correct a disease-causing error that was inherited or crept into our DNA when it replicated. Or, in some cases, we may want to enhance the genetic code of crops, livestock or perhaps even people.

So do we just snip the unwanted gene out and replace it with a good one?

Read more: Explainer: what is genome editing?

We first have to remember that animals and plants are composed of millions of cells, and each cell contains the same DNA. There is no point editing just one cell: we would have to edit the same gene in every single cell. Wed have to snip out millions of genes and paste in millions of new ones.

And not all cells are easy to get to how could we reach cells buried in our bones or deep within a brain?

A better approach is to start at the beginning and edit the genome while there is only one cell a very early embryo.

So, all we need is a giant microscope and a tiny pair of scissors. And that is basically what we use.

Cas9 is the technical name for the virus-destroying scissors that evolved in bacteria. The CRISPR part of the name comes from repeat DNA sequences that were part of a complex system telling the scissors which part of the DNA to cut.

In order to target our Cas9 scissors, we link them to an artificial guide that directs them to the matching segment of DNA.

Remember, DNA comes in two strands, with one strand fitting alongside the other. We make a guide with a code that will line up with only one part of our 3 billion base pair long genome its like a Google search. Its truly possible for our guide to comb through vast amounts of genetic material to find the one section it matches exactly. Then our scissors can make the cut in exactly the right place.

Once the Cas9 scissors cut the DNA just where we intend, the cell will try to repair the break using any available DNA it can find. So, we also inject the new gene we want to insert.

Read more: Now we can edit life itself, we need to ask how we should use such technology

You can use a microscope and a tiny needle to inject the CRISPR/Cas9 together with the guide and the donor DNA, the new gene. Or, you can punch holes in cells with electric currents and let these things just float in, use guns to shoot them in stuck-on tiny bullets, or introduce them encapsulated in bubbles of fat that fuse with the cell membrane and release their contents inside.

But how does the new gene find the right place to embed itself? Imagine you wanted to put in the last piece of a jigsaw puzzle with 3 billion pieces, and its inside a cell, filled with goop like a passionfruit.

What youd do is fabricate a jigsaw piece of precisely the right shape and inject it into the passionfruit. Then its just a case of jiggling around until eventually the piece finds its way to the correct part of the puzzle and slots into the only place it fits.

You dont need to be able to see the DNA in our genome through the microscope its too small. And you dont really have to jiggle either random diffusion (called Brownian motion) will always deliver the jigsaw piece to the place where it fits in the end.

First, the guide will jiggle along and find the right place for the scissors to cut, and then the new donor DNA will similarly line up where it fits and will be permanently stitched into the DNA strand via natural DNA repair mechanisms.

Recently, though, new CRISPR editing systems have been created that dont even require a cut through the DNA. In this case, the CRIPSR/Cas and guide system can deliver an enzyme to a particular gene and alter it, changing perhaps an A to a G or a C to a T, rather than cutting anything out or putting anything in.

Most experiments use mouse embryos or cells grown in petri dishes in artificial liquid designed to be like blood. Other researchers are modifying stem cells that may then be re-injected into patients to repopulate damaged organs.

Only a few labs around the world are actually working with early human embryos. This research is highly regulated and carefully watched. Others work on plant cells, as whole plants can be grown from a few cells.

As we learn more, the scope of what we can do with CRISPR/Cas9 will improve. We can do a lot, but every organism and every cell is different. Whats more, everything in the body is connected, so we must think about unexpected side effects and consider the ethics of changing genes. Most of all we, as a society, should discuss and agree what we wish to achieve.

Read more: Why we can trust scientists with the power of new gene-editing technology

Read the other articles in our precision medicine series here.

Read more here:
What is CRISPR gene editing, and how does it work? - The Conversation

Recommendation and review posted by Bethany Smith