The death of Lenin - what Vladimir Ilyich really died from. Cause of death

Soviet fighter pilot Valentin Bondarenko could become the first man in space. In any case, he had a chance of this... But he simply did not live to see his first flight: he died by a stupid accident during testing.

First squad

Valentin Vasilyevich Bondarenko was born on February 16, 1937 in Kharkov. His father, the head of a workshop at the Kharkov fur factory, went to the front in the first days of the war. Together with his mother and older brother, Valentin survived the German occupation. As a high school student, he began studying at the Kharkov flying club. In 1954, after graduating from school, he entered the Voroshilovgrad Military Aviation School, after its disbandment he transferred to Grozny, and then to the Armavir School, from which he graduated with honors in 1957. Bondarenko served in the aviation units of the Air Force of the Baltic Military District.

On April 28, 1960, Valentin’s cherished dream came true: after careful selection, he was enrolled in the first squad of Soviet cosmonauts. Out of several thousand applicants, only 29 people were selected.

Initially, Valentin was not one of the six candidates for space flight on the Vostok spacecraft. But for various reasons, several selected future cosmonauts dropped out of action, and Bondarenko was brought in for training.

Ridiculous death

The tests included a ten-day stay in a pressure chamber, the purpose of which was to test the reaction to the absence of external stimuli. It was believed that the conditions in the chamber were close to the conditions inside the spacecraft. It was located at the Air Force Research Institute-7 (now the Institute of Aviation and Space Medicine).

At the end of the experiment, Valentin was informed that he could remove the medical sensors attached to his body. There were red marks left at the attachment points, which Bondarenko wiped with a cotton swab soaked in alcohol. After that, the young man, without looking, threw the cotton wool towards the trash can. But by an unfortunate accident, it landed on the spiral of a hot electric stove and immediately burst into flames... Since the chamber was filled with almost pure oxygen, the flame quickly spread. Bondarenko's woolen training suit caught fire.

It was not possible to open the chamber quickly due to the large pressure drop. When it was finally opened, the cadet was still alive. Doctors at the Botkin Hospital fought for his life for 8 hours. He died on March 23, 1961, just 19 days before Gagarin’s flight, who, together with his squad comrades, spent several hours in his hospital... The cause of death was burn shock.

Secret Hero

The death of Senior Lieutenant Valentin Bondarenko was not reported anywhere: in those days, everything related to space was strictly classified. However, on June 17, 1961, by decree of the Presidium of the Supreme Soviet of the USSR, Bondarenko was posthumously awarded the Order of the Red Star “for the successful completion of the government’s assignment.”

At Bondarenko’s grave, located in Kharkov at the Filippovsky cemetery, an obelisk was erected with the inscription: “In blessed memory from fellow pilots.” Only in the 80s did the postscript appear: “-cosmonauts of the USSR.”

Bondarenko left behind his wife Anna and son Alexander. For some time they continued to live in Star City, where Anna worked at the Cosmonaut Training Center, then they left for Kharkov, where they had relatives. Until Sasha’s 16th birthday, he was paid a pension of 100 rubles for his father - quite decent money at that time. Subsequently, Alexander Bondarenko followed in his father’s footsteps and became a military pilot.

Only in 1980 did the Western press begin to write about the death of Valentin Bondarenko. In the USSR, an article about him was first published in 1986 in Izvestia. In 1991, one of the lunar craters was named after Bondarenko, and in July 2013, the name of the cosmonaut was given to school No. 93 in the city of Kharkov, where he once studied.

It would seem that the death of Valentin Bondarenko has nothing to do with the space flights themselves - it is simply a tragic accident. However, without such mistakes and tragedies, astronautics could not develop. By the way, this story forced engineers and scientists to reconsider the design of the test pressure chamber, in particular, changing the composition and pressure of the atmosphere, which was taken into account in the further development of manned spacecraft.

Today, in the Museum of the First Flight, located in the city of Gagarin, Smolensk region, in the small homeland of the world’s first man to go into space, you can see a deaf chamber, which is an exact copy of the one in which Valentin Bondarenko died.

It is obvious that after the death of the USSR and socialism, the death of Russia itself is inevitable. It is the matter of time. The West, which destroyed the USSR with the help of traitors from the top of the Politburo of the CPSU Central Committee, will destroy Russia. By the way, also with the help of traitors: the Yeltsins, Chubais, Gaidars, Putins, Medvedevs, Nemtsovs, Yavlinskys and other Navalnys. It will destroy because Russia in its current form, large and numerous with nuclear weapons and missiles, is the main geopolitical enemy in the world. And there is always a danger that Russia will throw off its power - the proxies of the West, who have ruled Russia since 1991, and begin its revival. Since the Russians still have genetic memory and a thirst for justice, they can again revive the USSR or its analogue. Therefore, the West not only wants to deprive Russia of missiles and nuclear weapons, but also to split it into parts that conflict with each other. How the Russian Federation and Ukraine are now in conflict with each other, but once they were one. . Therefore, the West, with the help of its proteges who have ruled Russia for 27 years, is doing everything to prevent Russia from being revived and continuing to degrade. Until quantitative degradation turns into qualitative degradation, that is, into decay. Which will happen under the control of the West, just as the collapse of the USSR occurred. In new regions, the right guys will be put in new positions, who will faithfully serve the West. If necessary, the West, at the request of the Kremlin, will send in its troops to control the process and pacify the natives, that is, us.

At the moment, science and industry have already been virtually destroyed in Russia. They made medicine inaccessible, which has completely degraded in 27 years. First of all, due to the decline in the quality of doctors. Since education in the country has completely degraded. All food products in supermarkets and convenience stores are counterfeit. All without exception. What does not add health to people. But the main thing is the destruction of science and industry. While they are still in the country. Thanks to the legacy of the USSR. But the legacy ends. And soon we will have to forget about space, missiles and nuclear weapons. The country will forget how to make them. The machine tool and electronics industries have already been destroyed. Factories producing bearings were destroyed. Factories producing high-tech products that produced watches, cameras, optics, domestic televisions, radios, refrigerators, and household appliances were deliberately bankrupted and closed. Russia produces mainly low-value products. And at the few remaining enterprises producing high value-added products, the share of imports exceeds 50%. Moreover, the most complex parts and assemblies: engines and their components, electronics. And the few parts that Russia produces, stamped iron, are produced using imported equipment. At AvtoVAZ in Tolyatti, the share of imports is 55% and continues to grow. At the plant producing Superjets, the share of imports is 75-80%. Moreover, almost all other aircraft factories in Russia were destroyed. The personnel training system for production was also destroyed. Old staff are retiring due to age, and there is no replacement for them. Over 27 years, about 80% of factories were closed. Most of it under Putin. And they keep closing. Hundreds of thousands of scientists have left Russia for the West over the years, since since 1992, science in the country has actually not been funded. Let me remind you that the Kremlin is doing all this deliberately on instructions from the West.

Here is what the wonderful Russian writer Valentin Rasputin wrote about this: “A state that deliberately kills itself - this has never happened in the world.” For me, everything has long been obvious, like twice two. Russia is essentially an occupied country that was defeated during the Cold War. Perestroika was a joint special operation by the CIA and the top of the CPSU to destroy the USSR and socialism. As a result of perestroika, the West was able to place its own agents at the head of our country, which were specially trained to govern Russia starting in the mid-70s. Prepared by traitors from the top of the CPSU Central Committee. At the head of the anti-Soviet conspiracy were Andropov and Kosygin. Head of the KGB and Prime Minister. Both are members of the Politburo. Traitors - the modern Russian elite is held tightly by the West by the balls, since all the money of the elite, trillions of dollars stolen from Russia, is stored in Western banks. We are ruled by the Vlasovites of the 21st century, ruled by the administration of the occupied territory, which does everything the West wants. No wonder the three-striped “Vlasov flag” proudly flies over the Kremlin. Controlling a colony with the help of a corrupt elite is a method long ago worked out by the Anglo-Saxons in their colonies. Particularly in India. By the way, Russia is now also a colony. And not only raw materials.

Now let’s assume that the West doesn’t care about Russia. That he doesn’t care what happens to her, will Russia be reborn then? No. But the process of death and decay will drag on. Why will this happen?

Capitalism in the world is divided into capitalism of metropolitan countries and peripheral capitalism. For capitalism to flourish in metropolitan countries, markets are needed, someone needs to be robbed, beads and mirrors need to be exchanged for natural resources, for cheap labor. Someone needs to snag dollars, euros, pounds sterling. He printed beautiful pieces of paper, the price of which is pennies, and with them you can buy whatever you want in the states of peripheral capitalism. Including the leadership of these countries. So that it implements in its states the economic policy that was prescribed in the metropolitan countries. Which is dictated by the IMF, the World Bank and the WTO - surrogates that created metropolitan countries so that the states of peripheral capitalism live by certain rules, according to which they will never be able to get out of hunger and poverty. For example, the countries of Latin America, in which capitalism is more than 150 years old, and they have not advanced one step towards the developed capitalism of the metropolitan countries.

The main wealth in the world is natural resources. And there are not enough of them for everyone to live well. It is the metropolitan countries that rob states with peripheral capitalism, taking away their raw materials through deception, buying them for pennies so that they can live beautifully themselves. The same USA is 20% of world GDP, and they consume 35% of world resources. Moreover, the laws of capitalism, which are good for metropolitan countries, are completely unsuitable for states with peripheral capitalism. But the latter, with the help of the IMF, the World Bank, and the WTO, force them to comply with the laws of capitalism that apply in the metropolitan countries. But the same Japan performed its “Japanese miracle” by violating these laws; the Japanese took the Soviet economy of the 30s as a model. But the Japanese were allowed to do this, which is why I will write below.

Sometimes the question arises: is there capitalism in Russia now? Yes, capitalism, typical peripheral capitalism. Which is fundamentally different from the capitalism of the metropolitan countries. I have already explained above why this happens. There are not enough resources for everyone. If Russia begins to live richly, then the West will have to live poorer and consume less. Let's take Moscow. Why is it fattening, its standard of living is the same as in the Czech Republic, and in the rest of Russia, like in Africa? So Moscow robs the entire country, lives off it, produces nothing, but the elite, owners of companies and factories live in it, pay taxes in it.

Similar relations are developing between metropolitan countries and states with peripheral capitalism. TNCs have taken over most of the enterprises and companies in the states of peripheral capitalism and are transferring all profits to the metropolitan countries. They invest in their countries, in their countries they pay taxes on the money they earned in Russia and other states of peripheral capitalism. That's where a lot of money and high salaries come from. Local businessmen do the same when they transfer money to the West. Many people pay taxes there.

Why is almost no money invested in business development in states of peripheral capitalism? Everyone in the world cannot live richly. The rich are rich because 90% are poor. Therefore, the states of peripheral capitalism are deliberately prevented from developing industry with the help of the IMF, the World Bank and the WTO. Which impose suicidal economic policies on these states. Therefore, in these states conditions have been deliberately created that make it unprofitable to create new enterprises. High loans, high taxes on literally everything, bribe-taking officials, racketeering. And in Russia, for example, there is also state racketeering if other obstacles do not work. The state just comes and takes your business away from you if you don’t understand it in an amicable way. So, in essence, they took away Euroset from Chichvarkin and Magnit from Galitsky, buying them for pennies. By the way, Magnit eventually went to Lavrov’s son-in-law. Who is our Minister of Foreign Affairs.

They rob us and do not allow us to be reborn. They do not give through political and economic methods. And sometimes they use military force against the dissatisfied, as well as conspiracies, sabotage, they organize counter-revolutions, “color revolutions”, coups: Chile - 1973, Arab countries of North Africa - 2010, Ukraine - 2014. And personal hostility has nothing to do with it. Just BUSINESS.

That is, Russia, as a country of peripheral capitalism, will simply not be allowed to develop, will not be allowed to emerge from a state of eternal degradation. And is it destined, like other states of peripheral capitalism, to degrade for centuries? No, it's not meant to be. What is not so dangerous for other states is deadly for Russia. Russia is an empire, the largest state in the world, home to hundreds of peoples. And all weak empires fall apart. This is an axiom. Just as the Great Ancient Empires of past centuries fell apart, when they weakened: Egypt, Greece, Carthage, Rome, the Ottoman Empire, Austria-Hungary, so Russia will also fall apart. The inevitability that miraculously did not happen in 1917, when Russia was saved by the Bolsheviks, will happen in the 21st century. Here is what the liberal and dissident Andrei Amalrik wrote about this in 1978: “Just as the adoption of Christianity delayed the death of the Roman Empire, but did not save it from the inevitable end, so the Marxist doctrine delayed the collapse of the Russian Empire - the third Rome - but was unable to avert it ". It may be objected that in 1991 the breakup had already occurred. I answer. The disintegration of the empire occurs until individual self-sufficient nations become separate states. That is, the collapse of 1991 was not final. Alas.

From the two parts I wrote above, it follows that in any situation under capitalism, Russia will have the same outcome - collapse. Who is to blame - I wrote. What to do? Only under socialism can Russia be raised from its knees. Therefore, we need to think about how to restore the USSR at a new stage. Is this real? No. The West has everything under control. He will not make the mistakes of 1917. He suffered too many troubles because of the October Revolution. Moreover, with the modern level of science and technology, television, it is much easier to control and zombie the masses. So it's too late to drink Borjomi. The collapse of Russia will happen in any case. At the same time, the Russians themselves will not find it enough. You have to pay for stupidity! The collapse of the USSR and the death of socialism could not be allowed. And now the train has left. But this does not mean that you have to stop fighting. You have to fight to the end in order to remain human, not to become a slave, a stupid chomping man in the street.

PS. Often, when I explain to people that states of peripheral capitalism will not be allowed to become metropolitan countries, they tell me about Japan and South Korea. And post-war Germany is sometimes cited as an example. The thing is that after WWII the Anglo-Saxons could not do without allies. The USSR was too strong and powerful. And socialism began to raise its head in Europe. The Anglo-Saxons needed an outpost that would prevent socialism from spreading further in Europe. Therefore, Germany was helped to revive with the help of the Marshall Plan. They opened their markets to Germany. Moreover, Germany became a colony of the Anglo-Saxons. There are tons of US military bases in it. Similarly, an outpost was needed in the east. It became another Anglo-Saxon colony - Japan. As a counterbalance to the USSR and China. It also has many US military bases. To boost the Japanese economy, an analogue of the Marshall Plan was used. The USA, England and other Western countries opened their markets for it. At the same time, Japan was allowed to use the experience of the USSR in the 30s. To accelerate the growth of your economy, to achieve a breakthrough, when in ten years you need to do what other countries took a century. They similarly helped South Korea, as an outpost in the confrontation with the DPRK. By the way, the standard of living in the DPRK in 1970 exceeded the standard of living in South Korea. In 1975, their standard of living was equal. In the second half of the 70s, stagnation began in the USSR. Which also affected the DPRK. Therefore, in 1980, South Korea managed to overtake the DPRK in terms of GDP per person. And after the collapse of the USSR and the death of socialism in Europe, the economy of the DPRK collapsed. Because the economy of North Korea was focused on the socialist countries in Europe and the USSR. And the West imposed an economic embargo on North Korea. Therefore, GDP per person in the DPRK in 1995 fell four times compared to 1987. North Korea's GDP fell accordingly by three times over the same period.

At all times, people have been interested in: why does a person die? In fact, this is quite an interesting question, to answer which we can consider several theories that can shed light on this situation. There are many different opinions on this topic, but in order to understand what death is and why a person is susceptible to it, it is necessary to uncover the mystery of old age. At the moment, a large number of scientists are struggling to solve this problem; completely different theories are being put forward, each of which, one way or another, has the right to life. But, unfortunately, none of these theories have been proven at the moment, and this is unlikely to happen in the near future.

Theories related to aging

As for opinions on the question “Why does a person die?”, they are all as diverse as they are similar. What these theories have in common is that natural death always comes with old age. A certain circle of scientists is of the opinion that old age as such begins at the moment of the emergence of life. In other words, as soon as a person is born, the invisible clock begins its reverse movement, and when the dial goes to zero, the person’s presence in this world will also cease.

There is an opinion that until a person reaches maturity, all processes in the body occur in the active stage, and after this moment they begin to fade away, along with this the number of active cells decreases, which is why the aging process occurs.

As for immunologists and some gerontologists who tried to find an answer to the question “Why does a person die?”, then, from their point of view, with age, autoimmune phenomena intensify in a person against the background of a decrease in the reaction of cells, which, in essence, leads to that the body’s immune system begins to “attack” its own cells.

Genetics, naturally, say that the whole problem lies in genes, while doctors argue that human death is inevitable due to body defects that accumulate throughout a person’s life.

Law of nature

Thanks to scientists from the USA who conducted research on this issue, it became known that people die while in the “kingdom of Morpheus”, mainly due to respiratory arrest. This occurs mainly in older people due to the loss of cells that control the breathing process, sending signals to the body to contract the lungs. In principle, such a problem can occur among a lot of people, its name is obstructive apnea, and this problem is the main one. But there cannot be such a cause of death as obstructive apnea. This is due to the fact that a person experiencing oxygen starvation (lack of oxygen) wakes up. And the cause of death is central sleep apnea. It should be noted that a person may even wake up, but still die due to lack of oxygen, which will result from a stroke or cardiac arrest. But, as mentioned earlier, this disease mainly affects older people. But there are also those who die before reaching old age. Therefore, a very reasonable question arises: why do people die young?

Death of the Young

It’s worth starting with the fact that recently, approximately 16 million girls in the age category from 15 to 19 years have become pregnant. At the same time, the risks of infant death are much higher than those of those girls who crossed the 19-year-old barrier. These problems are caused by both physiological and psychological factors.

Not the least reason is poor nutrition, and this is due to both obesity and problems associated with anorexia.

Smoking. Drugs. Alcohol

As for bad habits, such as abuse of alcohol, nicotine, and even more so drugs, this problem every year affects younger and younger segments of the population, who not only put their future children at risk, but also themselves.

Still, the most common cause of death among the young population is unintentional injuries. The reason for this can also be alcohol and drugs, not counting youthful maximalism, which cannot be discounted. Therefore, until teenagers reach adulthood, all responsibility for moral and psychological education lies entirely with the parents.

How does a person feel at the moment of death?

In fact, the question of a person’s feelings after death has worried all of humanity throughout its existence, but only recently have they begun to say with confidence that all people at the moment of death experience definitely the same feelings. This became known thanks to people who experienced clinical death. Most of them claimed that even lying on the operating table, being immobilized, they continued to hear and sometimes see everything that was happening around them. This is possible due to the fact that the brain is the last thing to die, and this happens mainly due to lack of oxygen. Of course, there are stories about a tunnel at the end of which there is a bright light, but there is virtually no reliability of this particular information.

Finally

Having delved into the problem and understood it, we can confidently answer the question: why does a person die? Quite often people ask themselves similar questions, but you should not devote your entire life to the problem of death, because it is so short that there is no time to spend it on understanding those problems for which humanity is not yet ready.

EXPERIMENTAL ARTICLES

UDC 577.15:576.367

Acadesine causes non-apoptotic death of tumor cells

V. A. Glazunova1*, K. V. Lobanov2, R. S. Shakulov2, A. S. Mironov2, A. A. Shtil1 "N.N. Blokhin Russian Oncology Research Center of the Russian Academy of Medical Sciences, 115478, Moscow, Kashirskoe sh. ., 24

■State Research Institute of Genetics and Selection of Industrial Microorganisms, 117545, Moscow, Dorozhny pr-d, 1 *E-mail: [email protected] Received by the editor December 27, 2012

ABSTRACT The effect of acadesine (5-aminoimidazole-4-carboxamide-1-0-O-ribofuranoside) on tumor and non-tumor cells of various species and tissue origin was studied. It has been established that acadesine causes non-apoptotic death of tumor cells; The sensitivity of non-tumor cells to the action of this compound is significantly lower. Acadesine causes the death of tumor cells with a drug resistance phenotype due to the expression of the P-glycoprotein transporter and inactivation of the proapoptotic protein p53. The activity of adenosine transporters is a necessary condition for cell death, whereas the function of AMP-activated protein kinase is not required. The predominant death of tumor cells under the influence of acadesine and the peculiarities of the mechanism of its cytotoxicity make this compound promising as an antitumor agent. Key words acadesine, cell death, tumor cells.

introduction

Acadesine (5-aminoimidazole-4-carboxamide-1-P-O-ribofuranoside, AICAR) is being tested in clinical trials as a drug for the treatment of chronic lymphocytic leukemia. An important property of acadesine is its primary toxicity to tumor cells with less pronounced damage to non-tumor cells. Previously, it was shown that acadesine is capable of stimulating AMP-activated protein kinase (AMPK), an important regulator of cell energy balance that controls the oxidation of fatty acids, glucose metabolism, and the synthesis of proteins, fatty acids and cholesterol. The mechanism of action of acadesine is due to its phosphorylation by adenosine kinase to form ZMP (5-amino-4-imidazolecarboxamide ribotide), an intermediate in the de novo synthesis of purine bases. ZMP, by mimicking the metabolic effects of AMP, is able to activate AMPK. The antitumor effect of acadesine is associated with the induction of apoptosis. At the same time, there is evidence of non-apoptotic cell death and an AMPK-independent mechanism of action of acadesine on tumor cells.

In this work, the effect of acadesine on mammalian cells was studied. It has been shown that acadesine causes the death of tumor cells of various tissues.

of new origin, including cells resistant to a number of antitumor agents. The mechanisms of cell death are different from apoptosis; Their important feature is the need for adenosine transport. Non-tumor cells are less sensitive to the action of acadesine. The selectivity of the cytotoxic effect and the specific mechanisms of tumor cell death may be important factors determining the prospects of using acadesine in tumor therapy.

experimental part

The following human cell lines were used in the experiments: HCT116 (colon adenocarcinoma), HCT116p53KO (isogenic subline in which p53 does not function), K562 (promyelocytic leukemia), K562/4 (subline obtained after selection for survival in the presence of doxorubicin; expressed multidrug resistance protein (MDR) P-glycoprotein; Pgp), MCF-7 (breast adenocarcinoma), MCF-7Dox (subline after selection for survival in the presence of doxorubicin; Pgp-mediated MDR phenotype), fibroblast culture PFC-2, blood lymphocytes from healthy donors, as well as mouse cells: P388 (lymphocytic leukemia) and Sp2/0 (myeloma). Reagents were purchased from PanEco, Russia (except where otherwise specified). Cells were cultured in mo-

Dulbecco's modified Eagle's medium with the addition of 5% fetal bovine serum (Bio-Whittaker, Austria), 2 mM L-glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin at 37°C, 5% CO2 in a humidified atmosphere. In the experiments, cultures in the logarithmic growth phase were used. Lymphocytes were isolated from the peripheral blood of donors by centrifugation in a Ficoll-urografin density gradient (d = 1.077 g/cm3).

Acadesine was obtained at GosNIIGenetika using a microbiological method using an original recombinant strain. In addition, the cytotoxicity of acadesine from Sigma was assessed. The same company purchased dipyridamole, an adenosine receptor inhibitor, 5-iodotubercidine, an adenosine kinase inhibitor that prevents the conversion of acadesine to ZMP, and zVAD-fmk (carbobenzoxyvalylalanyl-aspartyl-fluoromethylketone), a pan-caspase inhibitor. All compounds were dissolved in dimethyl sulfoxide or water (10-20 mM) and stored at -20°C. On the day of the experiment, dilutions of the drug were prepared in a culture medium. To assess the cytotoxicity of acadesine, we used the MTT test, staining of cells with propidium iodide and annexin V conjugated with fluorescein isothiocyanate (FITC), determination

cell cycle in flow cytometry and electrophoretic analysis of genomic DNA integrity. In some experiments, the reference drug was the alkyl cationic glycerolipid gas-P-(4-[(2-ethoxy-3-octadecyloxy)prop-1-yloxycarbonyl]butyl)-N-methylimidazolium iodide, an apoptosis inducer.

RESULTS AND DISCUSSION

Preferential sensitivity of tumor cells to acadesine

In preliminary experiments, we established that the microbiologically obtained acadesine preparation and commercial acadesine are identical in physicochemical properties, purity, storage stability and cytotoxicity (data not shown). For further studies, we used acadesine obtained by the author's method. In table. Figure 1 shows the cytotoxicity of acadesine for transformed and non-transformed cells (cultured or freshly isolated) of various species and tissue origin.

From the data presented in table. 1, it follows that the most sensitive to the action of acadesine are

Table 1. Cytotoxicity of acadesine for mammalian cells

Acadesine chains, mM

G G.125 G.25 G.5 1.G 2.G

K562 1GG* 1GG 70 46 9 G

P388 1GG 36 30 20 9 G

Sp2/0 1GG 34 29 14 G G

K562/4 1GG 1GG 72 42 8 G

MCF-7 1GG 1GG 82 50 15 2

MCF-7Dox 1GG 1GG 86 48 17 1

HCT116 1GG 1GG 50 36 23 G

HCT116p53KO 1GG 1GG 54 34 25 G

HPF-2, proliferating 1GG 1GG 1GG 96 96 86

HPF-2, non-proliferating** 1GG 1GG 1GG 1GG 95 92

Donor lymphocytes 1GG 1GG 1GG 98 94 90

Note. The results of the MTT test after 72-hour incubation of cells are presented. "The survival of cells incubated without acadesine was taken as 100%. Each value is the average of five independent experiments, standard deviation< 0%. ""Пролиферацию фибробластов останавливали культивированием клеток до монослоя (контактное торможение деления клеток).

P388 cells (mouse leukemia) and Sp2/0 (mouse myeloma): at an acadesine concentration of 0.125 mM, ~1/3 of the cell population survives. Other transformed cell lines studied also die under the action of submillimolar concentrations of acadesine. It is important that the cytotoxicity of acadesine is practically the same in the case of the K562 leukemic line and its subline with Pgp-mediated MDR (K562/4). The same is true for the MCF-7 breast adenocarcinoma line and the MDR subline (Table 1). A comparison of the cytotoxicity of acadesine against the HCT116 line and the HCT116p53KO subline (resistant to a number of DNA-damaging antitumor compounds) showed that inactivation of the proapoptotic p53 protein does not lead to an increase in cell survival in the presence of acadesine.

Equally important is the significantly higher survival of non-tumor cells in the presence of acadesine: the death of donor lymphocytes and non-transformed fibroblasts was practically absent even when exposed to acadesine in millimolar concentrations for 72 hours of continuous exposure (Table 1). Thus, acadesine causes preferential death of transformed cells (suspension and epithelial), including sublines resistant to other antitumor compounds. Non-tumor cells are damaged by acadesine to a much lesser extent. These features make it promising to use acadesine as an antitumor agent. However, for this it is important to establish the mechanisms of toxicity of acadesine to tumor cells.

Acadesine induces non-apoptotic cell death

The effect of acadesine on the distribution of ploidy of the colon adenocarcinoma cell line HCT116 was studied by flow cytometry. 24 hours after the addition of acadesine (0.25 mM), the accumulation of cells in the S phase was determined, and after 48 hours (Fig. 1), mass cell death was determined (the area to the left of the G1 peak; hypodiploid nuclei).

The accumulation of fragmented DNA may be a sign of apoptotic cell death if DNA cleavage occurs in the internucleosomal spaces, as evidenced by the formation of a set of fragments 140-170 bp long. with electrophoresis. To test this possibility, DNA integrity was determined in HCT116 cells treated with acadesine. It turned out that acadesine, unlike the reference drug - an alkyl cationic glycerolipid, does not lead to the appearance of a “ladder” of DNA fragments characteristic of apoptosis (Fig. 2).

Fluorescence

Rice. Fig. 1. Distribution of HCT116 cell line across phases of the cycle under the influence of 0.4 mM acadesine. A - intact cells; B - accumulation in the S phase after 24 hours; B - accumulation in the sub^1 area after 48 hours

An argument in favor of a non-apoptotic mechanism of death of HCT116 cells under the influence of acadesine is the results of cell staining with annexin U-FITC and propidium iodide (Fig. 3). Annexin U binds phosphatidylserine at the plasma membrane (translocation of phosphatidylserine from the inner lipid layer of the membrane

Rice. 2. DNA integrity in HCT116 cells.

1 - Intact cells;

2 - acadesine, 0.4 mM, 24 h;

3 - alkyl cationic glycerolipid, 6 µM, 24 h (method control)

Rice. 3. Staining of HCT116 cells with annexin V-FITC and propidium iodide. Pseudo-colors: red - intact cells; violet - acadesine (0.4 mM, 24 h); blue - alkyl cationic glycerolipid (method control; see caption to Fig. 2)

in the external is considered a sign of apoptosis). Propidium iodide is able to penetrate into cells undergoing necrosis (violation of the integrity of the plasma membrane). HCT116 cells treated with acadesine (0.4 mM, 24 h) did not stain with annexin V-FITC; on the contrary, the cells accumulated propidium iodide (Fig. 3), suggesting a necrotic component of the death mechanism. Similar results were obtained when registering necrotic cells using trypan blue (data not shown). Possibly, disruption of the integrity of the plasma membrane is a late event in acadesine-induced cell death. The reference drug, an alkyl cationic glycerolipid, induced an increase in annexin V-positive cells characteristic of apoptosis (Fig. 3).

Since apoptotic cell death suggests an active role for caspases, the effect of the pan-caspase inhibitor zVAD-fmk on acadesine cytotoxicity was studied. HCT116 cells were incubated with 200 μM zVAD-fmk for 30 min, after which acadesine was added to the cultures and incubation continued for 24 h. The presence of zVAD-fmk did not reduce cell death, which confirms the conclusion about the non-apoptotic mechanism of acadesine cytotoxicity.

Interaction with adenosine receptors is necessary for the death of tumor cells under the action of acadesine

Transfer of acadesine from the extracellular environment to cells can be carried out by adenosine transporters. We studied the effect of dipyridamole, an inhibitor of these transporters, on the cytotoxicity of acadesine in the P388 cell line. It turned out that in the presence of dipyridamole, cells are insensitive even to relatively high (up to 0.8 mM) concentrations of acadesine (Table 2).

To clarify the role of the acadesine-MP-AMPK metabolic pathway in the cytotoxicity of acadesine

Table 2. Cytotoxicity of acadesine in combinations with dipyridamole or 5-iodotubercidin

Exposure to Acadesine, mM

0 0.08 0.1 0.2 0.4 0.8

Acadezine 100* 79 З8 ЗЗ 20 18

Acadesine + dipyridamole, 5 μM 100 100 99 99 100 101

Acadesine + 5-iodotubercidin, 0.05 μM 100 76 Z9 Z1 22 16

*Survival (%) of P388 leukemia cells according to the MTT test after incubation for 72 hours.

(its phosphorylation by adenosine kinase to form ZMP and activate AMPK), cells were incubated with acadesine and the adenosine kinase inhibitor 5-iodotubercidin. The inhibitor did not affect the cytotoxicity of acadesine (Table 2). It follows from this that cell death in response to acadesine is not due to the formation of ZMP and activation of AMPK.

Thus, the study of the mechanisms of cytotoxicity of acadesine revealed a number of features indicating the non-trivial nature of the pharmacological effects of this compound. Acadesine causes the death of cultured tumor cells with a significantly less pronounced effect on non-tumor cells. Acadesine is toxic to cells with molecular determinants of drug resistance - Pgp expression and non-functioning p53. It is important to emphasize the non-apoptotic nature of tumor cell death under the influence of acadesine. These results allow us to regard acadesine as a unique reagent for studying the mechanisms of tumor cell death and a promising drug candidate.

The question remains open about the intracellular target of acadesine, the interaction with which causes the death of tumor cells. We have shown that the condition for cell death is the functioning of adenosine transporters, while activation of AMPK is not required. It is reasonable to assume that tumors expressing these adenosine transporters and receptors will be most sensitive to acadesine. The role of purine base transport in cell death is not well understood; analysis of differential expression of adenosine transporters and receptors in different types of tumors is required. It is likely that increased expression of these molecules will be a new molecular marker of tumor sensitivity to acadesine and a criterion for selecting patients for appropriate therapy.

The work was supported by the Ministry of Education and Science of the Russian Federation (State contract No. 16.N08.12.1010), and was also partially supported by the Dynasty Foundation for Non-Profit Programs.

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Sudden death occurs as a result of a fast-flowing latent or clinically pronounced painful condition. As medical practice shows, sudden death in adults often occurs due to acute coronary insufficiency, congenital or acquired cardiac and vascular pathologies. Find out what symptoms may indirectly indicate a hidden threat.

What is sudden death

According to international medical recommendations, a person’s death within 6 hours after the appearance of the first symptoms of a pathological condition is considered sudden. Instant death, or translated into English sudden death, occurs without a known cause. In addition, there are no morphological signs on the basis of which an appropriate diagnosis of the patient’s sudden death can be made at autopsy.

However, during a post-mortem examination of a person, a pathologist, having compared all available data, can make a logical conclusion about the instantaneous or violent death of the person. In most cases, instant death is supported by changes in organs in which continuation of life for the shortest period of time is impossible.

Causes of sudden death

Statistics show that the main cause of most deaths is heart disease: ischemic pathology, the onset of ventricular fibrillation. At the same time, when answering what causes instant death, experts often name chronic illnesses that occur in a latent form for a long time, after which they suddenly worsen and lead to the unexpected death of a person. One of these deadly diseases is cancer.

In most cases, oncology develops asymptomatically and makes itself felt when the patient is often considered hopeless. Thus, malignant liver disease is the main cause of unexpected deaths in China. Another insidious disease that can lead to sudden death is AIDS, which claims millions of lives in Africa every year. In addition, it is worth mentioning separately about Mexico. This is the only country in which cirrhosis of the liver is the main cause of high mortality in the population.

In young age

Today, young men and women are exposed to the negative influence of modern lifestyle every day. From TV screens and the covers of fashion magazines, the cult of a slender (often dystrophic) body, accessibility and promiscuity is imposed on young people. Therefore, it is quite understandable that the mortality rate of people just beginning their life journey will increase over time. The main causes of instant death among boys and girls under 25 years of age are considered to be:

alcohol;
smoking;
promiscuity;
drug addiction;
malnutrition;
psychological sensitivity;
hereditary diseases;
severe congenital pathologies.

In a dream

Unexpected death in this condition occurs due to the loss of special cells responsible for the contractility of the lungs. Thus, scientists from the USA were able to prove that people die in their sleep in most cases due to central sleep apnea. In this case, a person may even wake up, but still leave this mortal world due to oxygen starvation caused by a stroke or cardiac arrest. As a rule, elderly people are susceptible to this syndrome. There are no specific treatments for central sleep apnea.

Sudden infant death

This syndrome was first described in the early 60s of the last century, although cases of instant death of infants were recorded earlier, but they were not subjected to such a thorough analysis. Young children have very high adaptive abilities and incredible resistance to a variety of negative factors, which is why the death of an infant is considered an exceptional situation. However, there are a number of external and internal reasons that can lead to sudden child death:

prolongation of the Q-T interval;
apnea (the phenomenon of periodic breathing);
deficiency of serotonin receptors;
overheat.

Risk factors

Due to the fact that the main cardiogenic cause of instant death is ischemic disease, it is quite logical to assume that the syndromes accompanying this heart pathology can be fully attributed to conditions that can increase the likelihood of sudden death. With all this, it has been scientifically proven that this connection is mediated through the underlying disease. Clinical risk factors for the development of clinical death among patients with ischemic syndrome are:

acute myocardial infarction;
post-infarction macrofocal sclerosis;
unstable angina;
heart rhythm disturbance due to ischemic changes (rigid, sinus);
ventricular asystole;
myocardial damage;
episodes of loss of consciousness;
damage to the coronary (heart) arteries;
diabetes;
electrolyte imbalance (eg, hyperkalemia);
arterial hypertension;
smoking.

How does sudden death occur?

This syndrome develops in a matter of minutes (less often hours) without any warning in the midst of complete well-being. In most cases, instant death affects young men aged 35 to 43 years. Moreover, often during the pathological examination of the deceased, vascular causes of sudden death are discovered. Thus, studying the increasing cases of instant death, experts came to the conclusion that the main provoking factor in the occurrence of this syndrome is a violation of coronary blood flow.

For heart failure

In 85% of cases, immediate death is recorded in individuals with structural abnormalities of the organ that pumps blood into the vessels. In this case, sudden cardiac death looks like a lightning-fast clinical variant of coronary disease. Medical practice shows that in a quarter of people who die instantly, bradycardia and episodes of asystole are observed before the onset of primary symptoms. Death from cardiac arrest occurs due to the launch of the following pathogenetic mechanisms:

Reducing left ventricular fractional ejection by 25-30%. This syndrome greatly increases the risk of sudden coronary death.
Ectopic focus of automatism in the ventricle (more than 10 ventricular extrasystoles per hour or unstable ventricular tachycardia), arising as a consequence of ventricular arrhythmias. The latter mostly develop against the background of acute transient myocardial ischemia. An ectopic focus of automatism is usually classified as a risk factor for sudden arrhythmic death.
The process of spasm of the blood vessels of the heart, which leads to ischemia and contributes to the deterioration of the restoration of blood flow to damaged areas.

It is worth noting that tachyarrhythmia is a particularly significant electrophysiological mechanism resulting in sudden coronary death in a person with heart failure. At the same time, timely treatment of this condition using a defibrillator with a modified pulse configuration significantly reduces the number of deaths among patients who have suffered sudden cardiac arrest.

From a heart attack

Blood enters the heart through the coronary arteries. If their lumen closes, primary foci of necrosis and ischemia form in the heart. Acute manifestation of cardiac pathology begins with damage to the vascular wall with further thrombosis and spasm of the arteries. As a result, the load on the heart increases, the myocardium begins to experience oxygen starvation, which affects its electrical activity.

As a result of a sudden coronary spasm, ventricular fibrillation occurs, a few seconds after which a complete cessation of blood circulation to the brain occurs. At the next stage, the patient experiences respiratory arrest, atony, and absence of corneal and pupillary reflexes. After 4 minutes from the onset of ventricular fibrillation and complete cessation of blood circulation in the body, irreversible changes occur in the brain cells. In general, death from a heart attack can occur in 3-5 minutes.

From a blood clot

In the venous bed, these pathological formations arise due to the uncoordinated work of the coagulation and anticoagulation systems. Thus, the onset of the appearance of a clot is caused by damage to the vascular wall and its inflammation against the background of thrombophlebitis. Perceiving the appropriate chemical signal, the coagulation system comes into action. As a result, fibrin threads form near the pathological area, in which blood cells become entangled, creating all the conditions for the blood clot to break off.

In arteries, the formation of clots occurs due to narrowing of the vascular lumen. Thus, cholesterol plaques block the path of free blood flow, resulting in the formation of a lump of platelets and fibrin threads. It is important to note that in medicine a distinction is made between floating and mural thrombi. Compared to the first type, the latter has a slight chance of breaking off and causing a blockage (embolism) of the vessel. In most cases, the causes of sudden cardiac arrest from a blood clot are due to the movement of a floating thrombus.

One of the serious consequences of the separation of such a clot is blockage of the pulmonary artery, which is expressed in a strong cough and bluish skin. Often there is a violation of breathing with subsequent cessation of cardiac activity. An equally serious consequence of the separation of a thrombus is a violation of cerebral circulation against the background of embolism of the main vessels of the head.

Diagnosis of sudden death

A timely physical examination is the key to the success of further cardiopulmonary resuscitation (CPR) measures. Diagnosis of instant death is based on symptoms characteristic of the patient's natural death. Thus, absence of consciousness is determined if no external stimuli cause reactions on the part of the person being resuscitated.

Diagnosis of breathing disorders is noted when within 10-20 s. observation fails to detect coordinated movements of the sternum and the noise of the air exhaled by the patient. In this case, agonal breaths do not provide adequate ventilation of the lungs and cannot be interpreted as spontaneous breathing. During ECG monitoring, pathological changes characteristic of clinical death are detected:

ventricular fibrillation or flutter;
cardiac asystole;
electromechanical dissociation.

Clinical manifestations

In 25% of cases, sudden death occurs instantly without any warning signs. Some patients, a week before clinical death, complain of various prodromal manifestations: increased pain in the sternum, general weakness, shortness of breath. It is important to note that today there are already methods for preventing heart attacks based on early diagnosis of the warning symptoms of this condition. Immediately before the onset of sudden death, half of the patients experience an anginal attack. Clinical signs of a patient’s imminent death include:

loss of consciousness;
absence of pulse in the carotid arteries;
pupil dilation;
lack of breathing or the appearance of agonal breaths;
change in skin color from normal to gray with a bluish tint.

Medical care for sudden death

Typically, most cases of unexpected cardiac arrest occur outside the hospital. For this reason, it is extremely important to master the technique of providing emergency care in case of sudden clinical death. This is especially true for subjects of society who, due to their job responsibilities, come into contact with a large number of people. Remember, competent resuscitation actions immediately in the first minutes after the onset of symptoms of cardiac arrest will help gain time until medical workers arrive.

Urgent Care

The main problem that arises in unconscious persons is obstruction of the airways by the root of the tongue and the epiglottis due to muscle atony. It must be said that this condition develops in any position of the body, and when the head is tilted forward, it develops in 100% of cases. Therefore, the first thing that needs to be done is to ensure proper airway patency. For this purpose, you need to use P. Safar’s triple technique, consisting of the following sequential actions:

Throwing back the head;
Moving the lower jaw forward;
Opening the mouth.

Once airway patency is ensured, you should proceed to artificial pulmonary ventilation (ALV). When providing first aid, this activity is carried out using the mouth-to-mouth method. So, one hand is placed on the victim’s forehead, while the other pinches his nose. Then the resuscitator fixes his own lips around the mouth of the person being revived and blows air, while controlling the excursion of the patient's chest. When it is visible, you need to release the victim’s mouth, giving him a chance to exhale passively.

At the next stage, artificial maintenance of blood circulation is carried out, to ensure which an algorithm for performing indirect cardiac massage or chest compression is used. For this purpose, you need to correctly lay the person being resuscitated on a flat surface. Next, you should determine the compression points: by palpating the xiphoid process and moving away from it 2 transverse fingers upward.

The hand must be placed on the border of the middle and lower part of the sternum so that the fingers are parallel to the ribs. Pushes are performed with the limbs straightened at the elbows. Chest compression is performed at a frequency of 100 compressions per minute with a break for artificial ventilation. The depth of the shocks is about 4-5 cm. Measures to restore cardiac activity should be stopped if:

A pulse appeared in the main arteries.
The actions taken do not have the desired effect within 30 minutes. The exception is the following conditions that require prolongation of resuscitation:

hypothermia;
drowning;
drug overdose;
electrical injury.

Resuscitation measures

Today, the concept of CPR is based on strict rules that ensure complete safety of the activities carried out for human life. In addition, an algorithm for the resuscitator’s actions in case of sudden cardiac arrest or sudden loss of respiratory function in the injured person is presented and scientifically substantiated. In the development of these conditions, time plays a major role: only a few minutes separate a person from death. The algorithm for performing cardiopulmonary resuscitation involves performing the following actions:

Determining the condition of the victim, on the basis of which the range of measures necessary for revival is selected;
Early initiation of CPR, which involves performing two manipulations: chest compressions and artificial ventilation.
If the second stage is ineffective, they proceed to defibrillation. The procedure involves applying an electrical impulse to the heart muscle. In this case, direct current discharges should be applied only if the electrodes are correctly positioned and have good contact with the victim’s skin.
At this stage, as a rule, the victim is provided with specialized medical care, including the following early treatment measures:

artificial ventilation with tracheal intubation;
drug support, involving the use of:
catecholamines (adrenaline, atropine);
antidiuretic hormones (Vasopressin);
antiarrhythmic drugs (Cordarone, Lidocaine);
fibrinolytic agents (Streptokinase).
intravenous drip administration of electrolyte or buffer solutions (for example, sodium bicarbonate is administered for acidosis)