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CRISPR Gene Therapy Pain Relieves Better than Opiates

In 2006, a case of a rather rare genetic disease was described. One boy, originally from Pakistan, showed a very interesting deviation: he could feel the warmth, cold, texture of objects, easily recognize touch, but did not feel pain. Unfortunately, this boy is no longer alive, since he believed in his “invulnerability” so much that he jumped from the roof in an attempt to prove it. However, a group of experts managed to conduct some research on the genome of a young Pakistani, and these data today will help to createpainkillers that will act at the genetic level. They will work better than opiates and will not be addictive.

How to relieve pain

As you know, in order to develop such an anesthetic, scientists had to use the CRISPR genome editor . A new approach to pain relief imitates a rare mutation of a Pakistani boy. And he’s already tested on mice in the Prashant Mali lab at the University of California at San Diego. 

A study in which CRISPR was usedto temporarily block the pain, it took place in July, but the scientists did not want to give any comments until all the data of their research passed a full check. The main idea is to introduce virus particles into the cerebrospinal fluid that carry a modified version of CRISPR designed to interrupt pain signals. Modification (or rather temporary blocking) is subjected to a gene called SCN9A, which is responsible for the production of a particular molecule (it was called Nav1.7). It, in turn, is a key factor in the passage of a pain impulse into the brain.

This is interesting: Using CRISPR, for the first time, it was possible to edit several genes simultaneously.

Evidence of the dominant role of this gene and its corresponding molecule was obtained from people with strange hereditary diseases that cause a decrease in the pain threshold. The Pakistani boy already mentioned at the beginning of the article was a member of a family with a mutation that completely turned off the SCN9A gene. Thanks to kollingrx.com, it was possible to study this mutation quite well. But so far, attempts to mimic this mutation have not been successful, but researchers from California have succeeded.

The data obtained during experiments on animals suggest that it is not yet possible to completely remove the pain syndrome. But to significantly reduce its manifestation is already quite capable. The effect of CRISPR therapy is comparable to the use of opiates. The only exception is that CRISPR is not addictive. This is a huge step forward.

Innovation: CRISPR Gene Therapy Pain Relieves Better than Opiates
Computer simulation of how CRISPR works when DNA fragments are “cut out”

At the moment, scientists plan to start full-fledged research involving people. Experts say that such gene therapy will help to relieve a huge number of people from chronic pain. For example, patients in advanced stages of cancer or people with congenital neuralgia. Also, one cannot exclude the more “standard” use of a new type of pain medication: the use of CRISPR drugs during operations or in patients with serious injuries, for example, after accidents or in war zones. 

For faster entry into the market, scientists have created the company Navega Therapeutics, which will work with the FDA (U.S. Food and Drug Administration) and a number of research centers to test the health of the new drug. 

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Superinfection, antibiotics and apocalypse models

Bacteria, not cancer, terrorists and disasters claimed the lives of a huge number of people. Yersiniapestisalone (plague wand) resulted in the death of 150 million people. Taking antibiotics is the main method of combating bacterial infections. But what if the drugs stop working?

This is not a rhetorical question and not an introductory one for modeling a hypothesis, but a real situation in the foreseeable future against the background of increased bacterial resistance. The World Health Organization (WHO) and national services have a common plan of action, but no state has a detailed algorithm for preventing a bacterial pandemic.

We will talk about the causes of the impending catastrophe, as well as about attempts to find a way out of the medical impasse with the help of related disciplines – genomics, mathematics and virology.

Against life

Knowledge is power !: Superinfection, antibiotics and apocalypse models
c )

Even in antiquity, doctors used to treat purulent wounds with moldy bread. Of the hundreds of potions that existed in ancient Egypt, stale bread was one of the few effective remedies – an antibiotic from mold did relieve inflammation.

Having no idea about the growth and development of microorganisms, the Egyptians acted blindly. In the Middle Ages, doctors completely forgot about successful practice and treated someone in what a great deal, especially fond of bloodletting.

Only at the end of the XIX century. several researchers independently began to study molds from the genus Penicillium , paving the way for the appearance of penicillin. It soon became clear that the substances obtained from the fungus are active against many pathogenic microorganisms.

The British bacteriologist Alexander Fleming announced the discovery of penicillin on September 30, 1928. In 1938, scientists isolated pure penicillin in Oxford, and already in 1941 the drug was used to treat bacterial infections. In 1945, Fleming received the Nobel Prize for his services in the field of medicine.

The action of the first antibiotics (literally from the Greek “anti” – against, “bios” – life) is based on the suppression of the synthesis of molecules involved in the construction of the outer cell membrane of the bacterium.

The video shows how bacteria (on the left side of the screen) literally explode when penicillin is added – antibiotic-thinned cell walls cannot contain intracellular pressure. Fortunately, the structure of the membrane of human and animal cells is different from the bacterial one.

Efficiency trap

Knowledge is power !: Superinfection, antibiotics and apocalypse models
The daily dose of antibiotics per 1 thousand people, distributed across countries with different income levels. 

The potential danger of antibiotic resistance, that is, bacterial resistance to antibiotics, was pointed out by Alexander Fleming himself, but for decades the therapy has been uncontrolled. The “heady efficacy” became the fault – new drugs treated bronchitis, otitis media, conjunctivitis, skin infections, pharyngitis, diarrhea and plague.

Today, antibacterial drugs defeat gastritis, sexually transmitted diseases and even some tumors . Accordingto WHO experts, in developed countries, antibiotics have added approximately 20 years to the average life expectancy of a person. It is not surprising that in America alone today they produceabout 17 thousand tons of antibiotics.

Humanity seemed to have set itself the goal of defeating all the microbes on Earth, but the battle in this world war is not in our favor. Antibiotics have been around for less than a century, and bacteria have been living on the planet for 3 billion years, and create new generations every 20 minutes.

In 2014, the project “Review of Antimicrobial Resistance”, created under the patronage of British Prime Minister David Cameron, published estimates of global losses from antibiotic resistance after 30 years: 10 million deaths annually is more than cancer.

US Center for Disease Dynamics report showsdisturbing results in the development of resistance to last-line drugs prescribed in the absence of a reaction to other drugs. As people increasingly take antibiotics uncontrollably, treatment in medical facilities is losing their effectiveness.

The reports from hospitals in all regions of the world, especially in developing countries, which indicate the occurrence of incurable or almost incurable infections, are very worrying. Simple diseases that were easily treated with affordable drugs in the past become fatal.

According to the WHO, antibacterial resistance in the XXI century. is one of the most serious threats to human health, food security and the global development of civilization.

Infectious foci

Knowledge is power !: Superinfection, antibiotics and apocalypse models
Distribution of methicillin-resistant Staphylococcus aureus (Methicillin-resistant Staphylococcus aureus, MRSA), the causative agent of complex diseases, such as sepsis and pneumonia, resistant to most antibiotics (in percent). 

Sustainability is a natural response of evolution, an attempt to find a way out of the impasse inherent in natural conditions not only for microorganisms. Weeds “learn” to survive under the attack of pesticides, insects are looking for a way to eat plants with a built-in gene of protein toxins, and a person fights pathogens with his mind – this powerful process is associated with the adaptation of all living things to each other.

The problem is that the length of the generation of bacteria is several days or hours, and a person lives for decades. We adapt too slowly. Even taking control over the course of evolution with the help of science, we did not take into account the human factor in the distribution of antibiotics, therefore:

  • the maximum possible dose is taken in a long course;
  • the drug is not used for its intended purpose (for the treatment of viral infections);
  • massive use of the same drugs is carried out without making a reserve.

More than 50% of antibiotics in many countries are used improperly. Among the “culprits” are both ordinary people who violate all conceivable instructions, and hospital staff, contrary to sanitary rules, contributing to the spread of infections between patients with open wounds and a weakened immune system.

Back in the XIX century. in hospitals, they chopped onions finely, killing some bacteria in polluted air. Today gloves, dressing gowns and masks, although mandatory protection, should be given a conditional “zero” level of security corresponding to the class of a medical institution in Central Africa.

A modern hospital can be equipped with HEPA filters (High Efficiency Particulate Arrestance) for air purification, unidirectional air distribution systems, ultraviolet disinfection lamps, and photocatalytic filters with natural oxidizing agents.

The hygienic requirements for the design, equipment and operation of hospitals, maternity hospitals and other medical hospitals in Russia are rather modest , but are still violated sometimes: the staff does not have time or forgets to sanitize the premises, although it is enough to use an ultraviolet lamp and bleach.

Millions of people are catching new infections in hospitals or taking antibiotics to treat the flu and colds. In addition, the use of antibacterial drugs in livestock and agriculture is growing.

Knowledge is power !: Superinfection, antibiotics and apocalypse models
The global consumption of antibiotics in livestock (in milligrams per 10 km²). 

In 2018, Roskontrol discovered antibiotics and antimicrobial agents in all tested chicken samples.

In the United States, approximately 80% of antibiotics are used by Fildena store: https://fildena.mobi. It is not surprising that antibiotics are associated with a high frequency of resistant bacteria in the intestinal flora of chickens, pigs and other food animals.

Antibiotics enter the bloodstream through a hospital dropper, in tablet form, along with a piece of steak, even from a fresh loaf of rye bread, since antibiotics are used to combat some plant diseases, and medicines consumed by animals enter the soil and plants with feces urine.

The threat affects everyone – globalization has made the world vulnerable to pandemics. People die from super infections that are resistant to all available antibiotics.

According to a report by the US Centers for Disease Control and Prevention, millions of people in America come into contact with antibiotic-resistant bacteria every year , and at least 23,000 die from infections.

On a global scale, the figure is amazing – 700 thousand victimsannually .

Mathematical models of epidemics

Knowledge is power !: Superinfection, antibiotics and apocalypse models
For 1990—2016. 146 models were created for the analysis of infections caused by the most common types of pathogens: HIV, flu, malaria, MRSA, tuberculosis (the size of the pie chart is proportional to the number of studies). 

Mathematicians and Data Science experts use special predictive models to develop new algorithms to deal with highly lethal rapidly spreading infectious diseases.

A few years ago, a student team of physicists at Leicester University (England) proposed one of these models to calculate the scale of the zombie epidemic.

Apparently, it will be difficult to survive in the apocalypse: 100 days after the outbreak of the epidemic, zombies that turn into undead daily with a 90% probability of at least one person will not be able to catch only 300 people around the world.

The basis of this study was the real epidemiological model of SIR, since 1927 used to describe the spread of diseases among the population. S → I → R , where S (t) is susceptible , the number of susceptible healthy (not yet ill) at time t , I (t) is infected , the number of patients at time t , R (t) is recovered , the number of immune to a disease (e.g., ill) at time t.

Also, the parameter N is sometimes used – number of people , the total number of people covered by the model: S (t) + I (t) + R (t) .

Knowledge is power !: Superinfection, antibiotics and apocalypse models
A sharp outbreak quickly depletes the susceptible pool. The days are indicated on the X axis, and the percentage of the population on the Y axis , where 1.0 = 100%. 

The model divides the population into infected, infected, and dead / recovered. SIR also takes into account the frequency of spread and extinction when individuals in a population come in contact with each other.

Knowledge is power !: Superinfection, antibiotics and apocalypse models
SEIR model with an 8-day incubation period. 

Researchers modified SIR many times . For example, a modelSEIR received the incubation parameter of infected but not yet infectious patients (Exposed population). SEIR is good at simulating outbreaks of tuberculosis caused by the bacterium Mycobacterium tuberculosis .

Knowledge is power !: Superinfection, antibiotics and apocalypse models
In this model, 17% of the population is constantly infected. 

SIS shows that the survivors do not acquire immunity, but return to the susceptible group. Because people remain susceptible after infection, the disease reaches a steady state in the population.

Fortunately, the distribution schedule for most resistant bacteria cannot be directly compared to the zombie epidemic model. In simulation models, most often do not take into account human behavior, its interaction with the environment and bacterial biology.

To create realistic antibiotic-resistant bacteria distribution schedules, millions of different scenarios must be taken into account – the process is remotely similar to modern meteorology and in terms of complexity will give odds to many well-known mathematical problems.

Knowledge is power !: Superinfection, antibiotics and apocalypse models
Simulation of the spread of resistant bacteria across Orange County, California: squares – no specific control measures; black lines – inconsistent control measures when threshold values ​​are reached; gray lines – coordinated control at threshold values. 

The illustration above is an example of a good model that calculates the likelihood of infection spreading to 28 hospitals and 74 nursing homes in a typical urban district. In the model, all infected people move around the healthcare ecosystem: they interact with doctors, nurses, beds, chairs and doors hundreds of millions of times.

The model shows that without enhanced control measures, including regular testing of patients for infection immunity, and quarantine for all carriers, the infection will circulate in the ecosystem on an ongoing basis – in almost every hospital for a decade.

When collecting data for models in the epidemiology of resistance, it will not be superfluous to consider antivirals. The vaccination movement arose shortly after the creation of the first smallpox vaccine by the British physician Edward Jenner . In 1885, 62 years after the explorer’s death, about 100 thousand. Protesters held a march, carrying banners antivaktsinatorskie, children’s coffins and effigies Jenner.

Nearly one and a half centuries have passed, and things are still there – recent outbreaksmeasles from New York to Moscow are associated with an exacerbation of the anti-vaccination campaign. Measles is an extremely contagious viral disease, but many vaccines are created against bacterial infections.

Knowledge is power !: Superinfection, antibiotics and apocalypse models
Map of infection in a biological simulator . 

Even game developers have joined in the fight against human stupidity. In the simulator Plague Inc. the player must infect the virus and destroy the earth’s population in a pandemic. In 2019, anti-vaccinators were added to the game. The result was a visual model: the anti-vaccination movement accelerates the spread of epidemics around the world.

Mathematics does not answer the question of how to resist an infection escaping from quarantine. Under the conditions of an epidemiological threat, the development of new methods of counteracting bacteria becomes a step of “last hope” – in the next part we will talk about attempts to find a way out of the bacteriological impasse.

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A fundamentally new cure for migraine will soon appear on the market. How does it work?

On May 17, the US Food and Drug Administration approved a new class of drug to prevent migraines in adults. The New York Times has figured out how he will conquer pain.

Migraine is a type of headache that greatly affects performance, reduces the quality of life and is found in 10% of people . With migraines, people may have a head split, they are often annoyed by light or sounds, many begin to feel sick. 

Now there are drugs that can relieve symptoms, as well as drugs that prevent seizures, but it cannot be said that they always work effectively. For example, it is believed that existing drugs for the prevention of migraine help halve the number of headache attacks in 50-75% of patients, but there are undesirable reactions.

The new drug Aimovig (active ingredient – erenumab) is recommended for people who have four migraine attacks per month or more. Studies have shown that erenumab is able to reduce the number of days with migraine by 1-2.5 days per month. The most common adverse reactions are discomfort at the injection site (the medicine is administered using a special Fildena site: https://myfildena.com/) and constipation. 

Pharmaceutical news: A brand new cure for migraine will soon appear on the market.  How does it work?
This is what Aimovig 
Amgen Inc. looks like / AP / Scanpix / LETA

Erenumab blocks the activity of the peptide, which plays an important role in the development of migraine. The fact that this peptide contributes to the onset of seizures became known back in the 1980s. Then the researchers tried to block it with the help of special molecules, but it turned out that such drugs are toxic. Therefore, scientists decided to use antibodies that block receptors for this peptide, that is, they did not allow it to affect the body. The idea turned out to be a good one, and now several companies are developing such drugs to combat migraines. 

Amgen and Novartis for the first time managed to get government approval for the free sale of such a drug. Antibodies need to be administered independently – once a month subcutaneously in the stomach, thigh or forearm.

The development of other companies is slightly different, and, for example, Alder is researching a drug that needs to be administered once every three months, but intravenously. It is estimated that by 2019, four such drugs will be approved in the United States. 

They are expensive – 6900 dollars a year. The fact is that the monoclonal antibodies that make up Aimovig are produced using living cells, which is a complex and expensive process. 

Novartis says the European Medicines Agency is already considering an application for approval of Erenumab. A decision is expected in the coming months

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Faecal microbiota transplantation prolongs the life of prematurely aging mice

Innovation: Transplantation of fecal microbiota prolongs the life of prematurely aging mice
Bárcena et al. / Nature Medicine, 2019

Transplantation of fecal microbiota and oral administration of bacteria Akkermansia muciniphila prolonged the life of mice with progeria (premature aging) and improved their health. Scientists resorted to such interventions after studying the intestinal microflora of mice and people with progeria and comparing it with the microflora of long-livers. Article published in Cenforce 100 Medicine – http://cenforceonline.net/

A number of health effects of intestinal microbiota are currently known.. So, in mammals, it affects the feeling of fullness, can protect against pathogens, is involved in the digestion and synthesis of vitamins, secondary bile and short chain fatty acids. Its association with obesity , type 2 diabetes , cardiovascular disease , non-alcoholic fatty liver disease , cancer, and the response to anti-cancer therapy is known . 

Previously, scientists investigatedthe profile of the intestinal microbiota in long-livers, but its changes in the syndromes of premature aging and the effect on such conditions have not been described. Progeria refers to very rare genetic disorders that show signs of aging at a very young age. The most studied Hutchinson-Guildford Progeria Syndrome (HCH) is inherited in an autosomal dominant manner. 

Clea Bárcena from the University of Oviedo in Spain and her colleagues set out to find out if there are specific dysbioses in the intestines that can be associated with progeroid syndromes, and what effect on mice with progeria may have changes in their microbiome.

First, scientists decided to figure out how relevant the indicator is the profile of the intestinal microbiota during progeria, whether it is possible to distinguish its characteristic features. To do this, they conducted studies on mice and humans. They compared the metagenomes of bacteria living in the intestines of LmnaG609G / G609G mice (SPHG model) and wild-type mice. In the first month of life, the data of both groups of mice were similar, but noticeable differences were found from the fourth. For example, in rapidly aging mice, the number of bacteria of the genera Akkermansia and Dehalobacterium decreased against the background of an increase in the number of proteobacteria and cyanobacteria. The scientists obtained similar results by repeating the study on mice Zmpste24 – / – (another model of CNG).

Further, the authors checked whether specific dysbiosis in the intestine is detected in patients with progeria. They studied the feces of four children with CNS and their healthy brothers and sisters, and also studied the fecal microbiome of a child with Nestor-Guillermo syndrome and his family. Despite the small number of samples, the results suggested that the environment has a greater effect on the intestinal microbiome than progeria. Further analysis showed a decrease in the number of bacteria of the genus Gemmige and an increase in the number of a number of clostridia. Scientists have concluded that dysbiosis worsens over time in both mice and people with progeria.

Scientists have suggested that people with exceptional lifespan may be carriers of a microbiome that can improve the health of others. Their findings are consistent with previous studies: in centenarians, they found large numbers of Akkermansia and Christensenellaceae bacteria and low levels of proteobacteria compared to controls. 

Further, scientists established whether mice with progeria will live longer if they change their microbiome. In this part of the work of Lmna-G609G micedivided into four groups of 11 individuals. Mice from the treatment group underwent transplantation of fecal microbiota (TFM) from healthy wild-type mice. In the control group were animals that were not subjected to any intervention. Mice of the third group received TFM from older LmnaG609G / G609G mice, and a microbiota transplant was simulated in the fourth group. 

Sick mice that received bacteria from healthy ones showed an improvement, progeria slowed down. This was manifested by a slowdown in weight loss, normalization of blood glucose levels, as well as a mitigation of some other symptoms. Most importantly, the mice from the treatment group lived on average 13.5 percent longer than the control (160 days versus 141 days).

Innovation: Transplantation of fecal microbiota prolongs the life of prematurely aging mice
The survival rate of LmnaG609G / G609G mice (red graph), Lmna-G609G mice after TFM from wild-type mice (green graph), and Lmna-G609G mice after TFM from old Lmna-G609G mice. 
Bárcena et al. / Nature Medicine, 2019

In the third group, transplantation did not lead to accelerated aging, but the mice showed metabolic disorders: they spent less energy and gained weight, they had higher blood glucose levels. Similar results were obtained on Zmpste24 – / – mice . 

A study of intestinal metagenome in both mice and humans has shown a decrease in the amount of A. muciniphila in premature aging syndrome. In addition, previously in animal experiments, the use of this bacterium positively affected the metabolism andimproved response to immunotherapy. Therefore, scientists found it advisable to check whether the appointment of A. muciniphila can improve the health of Lmna-G609G mice and prolong their life. The increase in life expectancy compared with the control in this case was slightly less than with TFM, on average, no more than 10 days. According to scientists, this indicates the protective role of the microorganism in accelerated aging. They explain this effect by increased expression of the Reg3g gene , thickening of the intestinal mucosa and improvement of its healing.

To clarify other possible mechanisms behind improving the health of mice and increasing their lifespan after TFM, scientists profiled the intestinal microbiota metabolome of wild-type mice, Lmna-G609G mice, and Lmna-G609G mice that received the transplant. After analyzing the differences, the authors suggested that enhancing the synthesis of secondary bile acids may play a positive role. 

Heidi Zapata from Yale University, which studied the microbiome of people with aging and age-related pathologies, calledthe study is interesting, but cautioned against prematurely projecting its results onto people. “I do not think that we are close to finding a pill that we could swallow. But given the importance of the microbiome, this portends future opportunities, ”she said. 

Earlier in a small study, scientists showed that oral administration of A. muciniphila in humans can reduce the risk of heart disease and diabetes in the metabolic syndrome. Scientists also described changes in the microbiome with fibromyalgia and demonstrated amelioration of autism symptoms in a small group of children who received TFM.