MXPA06004527A - Methods, compositions and devices for inducing stasis in cells, tissues, organs, and organisms - Google Patents

Abstract

The present invention concerns the use of oxygen antagonists for inducing stasis in cells, tissues, and/or organs in vivo or in an organism overall. It includes methods and apparatuses for achieving stasis in any of these biological materials, so as to preserve and/or protect them. In specific embodiments, therapeutic methods and apparatuses for organ transplantation, hyperthermia, wound healing, hemorrhagic shock, cardioplegia for'bypass surgery, neurodegeneration, hypothermia, and cancer is provided.

Description

METHODS, COMPOSITIONS AND DEVICES FOR THE INDUCTION OF STASYS IN CELLS, TISSUES, ORGANS AND ORGANISMS BACKGROUND OF THE INVENTION This application claims priority to the provisional patent application serial number 60 / 513,458, filed on October 22, 2003, provisional patent application serial number 60 / 548,150, filed on February 26, 2004, and provisional patent application serial number 60 / 557,942, filed June 8, 2004, which are incorporated herein by reference. The government may have rights in the present invention pursuant to grant number GM048435 of the National Institute of General Medical Sciences (NIGMS).

FIELD OF THE INVENTION The present invention relates in general to the field of cell biology. More particularly, it relates to methods and apparatuses for the induction "of stasis in cells, tissues, organs and organisms, using a substance that competes with oxygen.In certain modalities, there are methods and apparatuses for the treatment, prevention and diagnosis of diseases. and conditions in a subject exposed to an oxygen antagonist.

DESCRIPTION OF THE RELATED TECHNIQUE Stasis is a Latin term meaning "static." In the context of stasis in living tissues, the most common forms of stasis refer to the preservation of tissues for transplantation or reinsertion. Typically, said tissues are immersed in a physiological fluid, such as saline solution, and put in cold to reduce biochemical processes that lead to cellular damage. This stasis is incomplete, and can not be depended on for extended periods. In fact, the success of organ transplantation and the reinsertion of limbs is inversely related to the time that the organ or limb is out of contact with the intact organism. A more extreme version of stasis, includes the position of an entire organism in what is colloquially known as "suspended animation." Although still considered primarily within the realm of science fiction, it has achieved some notoriety when healthy individuals have been sought to be cryopreserved after death, in the hope that future medical discoveries will allow their rebirth, and cure their fatal diseases. Supposedly, more than a hundred people have been cryopreserved since the first attempt in 1967, and more than a thousand people have made legal and financial arrangements for cryonics with one of several organizations, for example, Alcor Life Extension Foundation. Such methods include the administration of anti-ischemic drugs, preservation at low temperature, and methods that perfuse whole organisms with cryosuspension fluids. However, it has not yet been proven that this form of organismal stasis is reversible. The utility of stasis induction in biological matter as contemplated by the compositions, methods or articles of manufacture described herein, is characterized by induction or stasis initiation, followed by a period in which stasis is maintained, followed then by reversion to a normal or quasi-normal physiological state, or a state that the person skilled in the art would recognize as a state that is better than the state than that of the biological matter that had never suffered stasis in whole or in part. Stasis can also be defined as what it is not. Organismal stasis is not any of the following states: sleep, comatose state, death, anesthesia or convulsion for great evil. There are numerous reports of individuals who have survived the apparent cessation of pulse and respiration after exposure to hypothermic conditions, usually in cold water immersion. Although not fully understood by scientists, the ability to survive such situations is probably derived from what is called the "immersion or dive reflection of mammals." It is thought that this reflex stimulates the vagal nervous system, which controls the lungs, heart, larynx and esophagus, to protect vital organs. Perhaps, the stimulation of nervous receptors on the skin by cold water, causes deviation of the blood towards the brain and towards the heart, and away from the skin, the gastrointestinal tract and the extremities. At the same time, protective reflex bradycardia, or delayed heartbeat, retains the diminished oxygen supplies within the body. Unfortunately, the expression of this reflex is not the same in all people, and it is thought that it is a factor in only 10 to 20% of the cases of immersion in cold water. Compositions and methods that do not completely or in any way depend on hypothermia and / or oxygen, may be useful in the context of organ preservation, as well as for the preservation of tissues or cells. Cells and tissues are commonly preserved using hypothermia, often at temperatures substantially below freezing, such as in liquid nitrogen. However, temperature dependence can be problematic, since apparatuses and agents for producing such low temperatures may not be readily available when needed, or may require replacement. For example, tissue culture cells are frequently stored for periods in tanks containing liquid nitrogen; however, these tanks frequently require that the liquid nitrogen in the unit be periodically replaced because otherwise it becomes depleted, and the temperature is not maintained. In addition, damage to cells and tissues occurs as a result of the freeze / thaw procedure. In this way, improved techniques are needed.

In addition, the lack of capacity to control the cellular and physiological metabolism in whole organisms subject to traumas such as amputation and hypothermia, is a key deficiency in the medical field. On the other hand, the anecdotal evidence discussed above strongly suggests that if properly understood and regulated, it is possible to induce stasis in entire cells, tissues and organisms. Thus, there is a great need for improved methods for the control of metabolic processes under traumatic conditions.

BRIEF DESCRIPTION OF THE INVENTION Therefore, the present invention provides methods, compositions, articles of manufacture and apparatus for inducing stasis in cells, tissues and organs located within an organism or derivatives thereof, as well as in the organism itself. Said methods, compositions, articles of manufacture and apparatuses can be used to protect biological matter, as well as to prevent, treat or diagnose diseases and conditions in the organism. Details of such applications and other uses are described below. The invention is based on studies with compounds that were determined to have a protective function, and thus, serve as protective agents. In addition, the general results of studies that include different compounds indicate that compounds with an available electron donor center are particularly effective in the induction of stasis. In addition, these compounds induce reversible stasis, meaning that they are not so toxic to the particular biological material that the material dies or decomposes. The present invention includes exposing biological matter to an amount of an agent, to achieve stasis of biological matter. In some embodiments, the present invention relates to methods for the induction of stasis in biological matter in vivo, comprising: a) identifying an organism in which stasis is desired; and b) exposing the organism to an effective amount of an oxygen antagonist to induce stasis in biological matter in vivo. The induction of "stasis" in biological matter means that matter is alive, but is characterized by one or more of the following: at least a double reduction in the rate or amount of carbon dioxide production by biological matter; at least a double reduction in the regime or amount of oxygen consumption by biological matter; and at least a 10% decrease in movement or mobility (applies only to cells or tissues that move, such as sperm cells or a heart or limb, or when stasis is induced in the entire organism) (referred to as a whole) as "cellular respiration indicators"). In the methods of the invention, stasis is temporary and / or reversible, meaning that the biological material does not exhibit the stasis characteristics any later in time. The term "biological matter" refers to any living biological material (mammalian biological material in preferred embodiments) that includes cells, tissues, organs and / or organisms, and any combination thereof. It is contemplated that stasis may be induced in a part of an organism (such as in cells, in tissues and / or in one or more organs), if that part remains within the organism or is removed from the organism, or the entire organism will be put in place. a state of stasis. The term "biological material in vivo" refers to biological material that is in vivo, that is, still inside or attached to an organism. In addition, the term "biological matter" will be understood as synonymous with the term "biological material". An organism that needs stasis is an organism in which the stasis of the whole organism, or part of it, can give direct or indirect physiological benefits. For example, a patient at risk of hemorrhagic shock may be considered to need stasis, or a patient who will undergo coronary artery bypass surgery may benefit from protection of the heart from ischemic / reperfusion injury. Other applications are discussed throughout the application .. In some cases, it is identified or determined that an organism needs stasis based on one or more tests, selections or evaluations that indicate a condition or disease, or the risk of a condition or disease that can be prevented or treated suffering from stasis. Alternatively, considering a patient's family medical or medical history (patient interview) can give information that an organism needs stasis.

The term "oxygen antagonist" refers to a substance that competes with oxygen as far as it is used by a biological material that requires oxygen to be alive ("biological material that uses oxygen"). Oxygen is typically used or required by several cellular processes that create the primary source of readily usable energy from biological matter. An oxygen antagonist effectively reduces or eliminates the amount of oxygen that is available for the biological material that uses oxygen, and / or the amount of oxygen that can be used by the biological material that uses oxygen. Thus, in some embodiments, an oxygen antagonist inhibits or reduces the amount of cellular respiration that occurs in cells, for example, by binding sites in the cytochrome c oxidase that would otherwise bind to oxygen. The cytochrome c oxidase binds specifically to oxygen, and then converts it to water. Preferably, the binding to cytochrome c oxidase by the oxygen antagonist is specific. In some embodiments, said binding to cytochrome c oxidase is preferably releasable and reversible binding (eg, it has an in vitro dissociation constant, Kd, of at least 10"z, 10" 3 or 10"4 M, and has an in vitro dissociation constant, Kd, not greater than 10"6, 10 ~ 7.10" 8, 10"9, 10" 10 or 10"11 M). In some embodiments, an oxygen antagonist is evaluated by measuring the production of ATP and / or carbon dioxide. The term "effective amount" means an amount that can achieve the established result. In the methods of the invention, an "effective amount" is, for example, an amount that induces stasis in biological matter that needs stasis. It will be understood that when stasis is induced in a tissue or organ, an effective amount is one that induces stasis in the tissue or organ, as determined by the collective amount of cellular respiration of the tissue or organ. Accordingly, for example, if the level of oxygen consumption by a heart (in conjunction with respect to heart cells) is decreased by at least about 2 times after exposure to a particular amount of a certain oxygen antagonist, it is You will understand that there was an effective amount that induced stasis in the heart. Also, an effective amount of an agent that induces stasis in an organism is one that is evaluated with respect to the collective or aggregate level of a particular stasis parameter. It will also be understood that when stasis is induced in an organism, an effective amount is one that induces stasis in general of the entire organism, unless a particular part of the organism is targeted. The concept of an effective amount of a particular compound refers to how much usable oxygen is available for biological matter. In general, stasis can be induced when there is approximately 100,000 ppm or less of oxygen in the absence of any oxygen antagonist (ambient air has approximately 210,000 ppm of oxygen). The oxygen antagonist serves to alter how much oxygen is actually available. In this way, while the actual concentration of oxygen at which the biological material is exposed may be greater, even much higher than 10 ppm, stasis may be induced due to the competitive effect of an oxygen antagonist with oxygen by binding to essential proteins. that metabolize oxygen in biological matter. In other words, an effective amount of an oxygen antagonist reduces the effective concentration of oxygen to a point where the oxygen that is present can not be used. This will occur when the amount of an oxygen antagonist reduces the effective oxygen concentration below the Km of oxygen binding to essential proteins that metabolize oxygen (ie, comparable to 10 ppm of oxygen). Accordingly, in some embodiments, an oxygen antagonist reduces the effective oxygen concentration by approximately 2., 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300 , 350, 400, 450, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600 , 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 5000, 6000, 7000, 8000, 9000 or 10000 times or more, or any scale derived in these numbers. It is understood that this is another way of indicating a decrease in cellular respiration. In addition, the effective amount can be expressed as a concentration with or without a rating in duration of exposure time. In some embodiments, it is generally contemplated that to induce stasis, the biological material is exposed to an oxygen antagonist by about, at least about, or at most about 5, 10, 15, 20, 25, 30, 35, 40 , 45, 50, 55, 60 seconds, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 , 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3 , 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 , 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more years, and any combination or scale derivable in these numbers. Then, the biological material may continue to be exposed to an oxygen antagonist or, in other embodiments of the invention, the biological material may cease to be exposed to the oxygen antagonist. This last step can be achieved by removing or effectively removing the oxygen antagonist from the presence of the biological material in which stasis was desired, or the biological material can be removed from an environment containing the oxygen antagonist. Therefore, in some embodiments of the invention, stasis is induced, and an additional step in the methods of the invention, is to keep the relevant biological matter in a state of stasis. This can be achieved by continuing to expose the biological matter to an oxygen antagonist, and / or by exposing the biological matter to a non-physiological temperature. Alternatively, the biological material can be placed in a preservative agent or solution, or it can be exposed to normoxic or hypoxic conditions. It is contemplated that the biological matter can be maintained in stasis for approximately at least approximately, or at most approximately 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45 , 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 , 11, 12 months, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more years, and any combination or scale derivable in these numbers. It will be appreciated that "stasis" with respect to an entire animal, and "stasis" with respect to cells or tissues, may require different durations of stasis time. Thus, with respect to human subjects, for example, subjects undergoing surgical treatment, treatment of malignant hyperthermia or who are victims of trauma, a stasis time of up to 12, 18 or 24 hours is generally contemplated. With respect to non-human animal subjects, for example, non-human animals sent or stored for commercial purposes, stasis is contemplated for a period of 2 or 4 days, 2 or 4 weeks, or more. The term "exposure" is used according to its ordinary meaning to indicate that biological matter is subjected to an oxygen antagonist. This can be achieved in some embodiments by contacting the biological matter with an oxygen antagonist. In the case of cells, tissues or organs in vivo"Exposure" can also mean "exposing" these materials, so that they can come into contact with an oxygen antagonist. This can be done, for example, surgically. The exposure of the biological material to an oxygen antagonist can be by incubation in or with (includes immersion) the antagonist, perfusion or infusion with the antagonist, injection of biological material with an oxygen antagonist, or application of an oxygen antagonist to the biological matter. In addition, if stasis of the whole organism is desirable, inhalation or ingestion of the oxygen antagonist, or any other route of pharmaceutical administration for use with the oxygen antagonists is contemplated. In some embodiments, an effective amount is characterized as a sublethal dose of the oxygen antagonist. In the context of the stasis induction of cells, tissues or organs (not the entire organism), a "sub-lethal dose" means a single administration of the oxygen antagonist that is less than half the amount of the oxygen antagonist that would make at least a majority of the cells in a biological material die within 24 hours of administration. If stasis of the whole organism is desired, then a "sub-lethal dose" means a single administration of the oxygen antagonist that is less than half the "amount of the oxygen antagonist that would cause the organism to die within 24 hours of administration. In other embodiments, an effective amount is characterized as an almost lethal dose of the oxygen antagonist.Also, in the context of the stasis induction of cells, tissues or organs (not the entire organism), an "almost lethal dose" means a single administration of the oxygen antagonist that is within 25% of the amount of inhibitor that would cause at least a majority of the cells to die within 24 hours of administration.If stasis of the whole organism is desired, then an "almost "lethal" means a single administration of the oxygen antagonist that is within 25% of the amount of the inhibitor that would cause the organism to die within 24 hours of the In some embodiments, a sublethal dose is provided by administering a predetermined amount of the oxygen antagonist to the biological material. In some embodiments, an effective amount is administered by monitoring, alone or in combination, the amount of oxygen antagonist administered, monitoring the duration of administration of the oxygen antagonist, monitoring a physiological response (e.g., pulse, respiration, pain response, movement or mobility, etc.) of the biological material to the administration of the oxygen antagonist, and reducing, interrupting or stopping the administration of the oxygen antagonist when a predetermined minimum or maximum is measured for a change in that response, etc. In addition, these steps can be used additionally in any method of the invention. In certain embodiments, the biological material is exposed to an amount of an oxygen antagonist that reduces the rate or amount of carbon dioxide production by the biological material, at least 2 times, but also by approximately, at least approximately , or at most approximately 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 200, 300, 400, 500 times or more, or any differentiable scale in these numbers. In other embodiments, the biological material is exposed to an amount of an oxygen antagonist that reduces the rate or amount of oxygen consumption by the biological material, at least 2 times, but also by approximately, at least approximately, or at most about 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 200, 300, 400, 500 times or more, or any Derivative scale in these numbers. In other modalities, biological matter is exposed to an amount of an oxygen antagonist that decreases movement or mobility by at least 10%, but also by approximately, at least approximately, or at most approximately 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99 or 100%, or any scale derivable in these numbers. As with other modalities, these characteristics and parameters are in the context of any biological matter that is induced in a state of stasis. In this way, if stasis is induced in the heart in an organism, these parameters would be evaluated for the heart, and not the entire organism. In the context of organisms, a reduction in oxygen consumption of the order of about 8 times, is a type of stasis referred to as "hibernation". Furthermore, it will be understood in this application that a reduction in oxygen consumption of the order of about 1000 times can be considered as "suspended animation". It will be understood that modalities of the invention can be achieved with respect to stasis, at the level of hibernation or suspended animation, if appropriate.

In addition, in some embodiments of the invention, methods are provided for the reduction of cellular respiration, which may or may not be as high as that necessary to reach stasis. A reduction in oxygen consumption by approximately, at least approximately, or at most approximately 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100%, is provided in the methods of the invention. It can also be expressed and evaluated in terms of any cell respiration indicator. It is contemplated that biological matter may be exposed to one or more oxygen antagonists more than once. It is contemplated that the biological material may be exposed to one or more oxygen antagonists 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more times, meaning when a biological material is exposed multiple times, that there are intermediate rest periods (with respect to exposure to the oxygen antagonist). In some cases, a sub-lethal collective dose or a non-lethal collective dose is administered to the biological material. As discussed above, with respect to the induction of stasis in biological matter that is not a whole organism, a "sub-lethal collective dose" means a quantity of multiple administrations of the oxygen antagonist, which together is less than half the amount of the oxygen antagonist that would cause at least a majority of the cells to die within 24 hours of one of the administrations. In other embodiments, an effective amount is characterized as an almost lethal dose of the oxygen antagonist.

Likewise, a "non-lethal collective dose" means a quantity of multiple administrations of the oxygen antagonist that is within 25% of the amount of the oxygen antagonist that would cause at least a majority of the cells to die within 24 hours of one of the administrations. It is contemplated that multiple doses may be administered to induce stasis in the entire organism. The definition for "sub-lethal collective dose" and "almost lethal collective dose" can be extrapolated on the basis of the individual doses initially discussed for stasis in whole organisms. It is contemplated that the biological material for use in the context of the invention includes any biological material comprising a cell that uses oxygen. The cell can be eukaryotic or prokaryotic. In certain modalities, the cell is eukaryotic. More particularly, in - some embodiments, the cell is a mammalian cell. Mammalian cells contemplated for use with the invention include, but are not limited to, those of: human, monkey, mouse, rat, rabbit, har, goat, pig, dog, cat, ferret, cow, sheep and horse. In addition, the cells of the invention can be diploid, but in some cases, the cells are haploid (sex cells). In addition, the cells can be polyploid, aneuploid or anucleated. The cell can be from a particular tissue or organ, such as one from the group consisting of: heart, lung, kidney, liver, bone marrow, pancreas, skin, bone, vein, artery, cornea, blood, small intestine, large intestine , brain, spinal cord, smooth muscle, skeletal muscle, ovary, testicle, uterus and umbilical cord. In addition, the cell can also be characterized as one of the following cell types: platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, skeletal muscle cell, endocrine cell, glial cell, neuron, secretory cell, cell with barrier function, contractile cell, absorption cell, mucosal cell, limb cell (cornea), stem cell (totipotent, pluripotent or multipotent), fertilized or unfecundated oocyte, or sperm. The biological material may be exposed to, or may be contacted with, more than, an oxygen antagonist. Biological matter can be exposed to at least one oxygen antagonist, including 2, 3, 4, 5, 6, 7, 8, 9, 10 or more oxygen antagonists, or any scale derivable in these numbers. With multiple oxygen antagonists, the term "effective amount" refers to the collective amount of oxygen antagonists. For example, the biological material can be exposed to a first oxygen antagonist, and can then be exposed to a second oxygen antagonist. Alternatively, the biological material can be exposed to more than one oxygen antagonist at the same time or in an overlapping form. In addition, it is contemplated that more than one oxygen antagonist may be included or mixed together, such as in an individual composition at which biological matter is exposed. Methods and apparatuses of the invention include a protective agent, which in some embodiments, is an oxygen antagonist. In other embodiments, the oxygen antagonist is a reducing agent. In addition, the oxygen antagonist can be characterized as a chalcogenide compound. In certain embodiments, the chalcogenide compound comprises sulfur, while in others, it comprises selenium, tellurium or polonium. In certain embodiments, a chalcogenide compound contains one or more sulfide exposed groups. It is contemplated that this chalcogenide compound contains 1, 2, 3, 4, 5, 6 or more exposed sulfide groups, or any scale derivable in these numbers. In particular embodiments, said sulfur-containing compound is CS2 (carbon disulfide). Furthermore, in some methods of the invention, stasis is induced in cells, exposing the cells to a reducing agent having a chemical structure of: where X is N, O, Po, S, Se or Te; where Y is N or O; wherein R- is H, C, lower alkyl, lower alcohol, or CN; wherein R2 is H, C, lower alkyl, or a lower alcohol, or CN; where n is 0 or 1; where m is 0 or 1; wherein k is 0, 1, 2, 3 or 4; and where p is 1 or 2. The terms "lower alkyl" and "lower alcohol" are used according to their ordinary meanings, and the symbols are those used to refer to chemical elements. This chemical structure will be referred to as the "reducing agent structure", and any compound having this structure will be referred to as a reducing agent structure compound. In additional embodiments, k is 0 in the reducing agent structure. In addition, in other embodiments, the Ri and / or R2 groups may be an amine or lower alkylamine. In others, R-, and / or R2 may be a short chain alcohol or a short chain ketone. In addition, R-i and R2 can be bridged, and / or the compound can be a cyclic compound. In other embodiments, X can also be a halogen. The term "lower" means that it refers to 1, 2, 3, 4, 5 or 6 carbon atoms, or any scale derivable in these numbers. In addition, Ri and / or R2 may be other small organic groups including, C2-C5 esters, amides, aldehydes, ketones, carboxylic acids, ethers, nitriles, anhydrides, halides, acyl halides, sulfides, sulfones, sulphonic acids, sulfoxides and / or thiols. Said substitutions are clearly contemplated with respect to Ri and / or R2. In some other embodiments, R-i and / or R2 may be short chain versions of the small organic groups discussed above. "Short chain" means 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon molecules, or any derivable scale in these numbers.

It is contemplated that the reducing agent structure compound may be a chalcogenide compound in some cases. In certain embodiments, the chalcogenide compound has an alkyl chain with an exposed chalcogenide. In others, the chalcogenide compound has a chalcogenide that becomes exposed once it is absorbed by the biological material. In this regard, the chalcogenide compound is similar to a prodrug as an oxygen antagonist. Therefore, one or more molecules of sulfur, selenide, oxygen, tellurium, polonium or unndhexium in the compound become available after exposure of the biological material to the chalcogenide compound. In this context, "available" means that sulfur, selenide, oxygen, tellurium, polonium or unndhexium will retain an electron. In other embodiments, the reducing agent structure compound is selected from the group consisting of H2S, H2Se, H2Te and H2Po. In some cases, the reducing agent structure has an X that is an S. In others, X is Se, or X is Te, or X is Po, or X is O. Additionally, k in the reducing agent structure is 0 or 1 in some modalities. In certain embodiments, the reducing agent structure compound is dimethyl sulfoxide (DMSO), dimethyl sulfide (DMS), carbon monoxide, methyl mercaptan (CH3SH), mercarptoethanol, thiocyanate, hydrogen cyanide, methanethiol (MeSH) or CS2. In particular embodiments, the oxygen antagonist is H2S, H2Se, CS2, MeSH or DMS. Particularly contemplated are compounds of! order of the size of these molecules (ie, within 50% of their average molecular weights). Furthermore, it will be generally understood that any compound discussed herein as an oxygen antagonist may be provided in the form of a prodrug to the biological material, meaning that the biological material or other substances in the environment of the biological material alter the prodrug in its active form, that is, in an oxygen antagonist. The oxygen antagonist is provided to the biological material in a state that allows it to compete with oxygen. The oxygen antagonist can be a gas, semi-solid liquid (such as a gel or paste), liquid or gas.

It is contemplated that the biological material may be exposed to more than one oxygen antagonist and / or an oxygen antagonist in more than one state. In certain embodiments, the oxygen antagonist is a gas. In particular embodiments, the gaseous oxygen antagonist includes carbon monoxide, nitrogen, sulfur, selenium, tellurium or polonium, or a mixture thereof. In addition, it is specifically contemplated that an oxygen antagonist is a chalcogenide compound as a gas. In some embodiments, the oxygen antagonist is in a gas mixture comprising more than one gas. The other gas or the other gases are a non-toxic gas and / or a non-reactive gas in some embodiments. In some embodiments, the other gas is a noble gas (helium, neon, argon, krypton, xenon, radon or unndoctium), nitrogen, nitrous oxide, hydrogen, or a mixture thereof.

In some cases, the gas mixture also contains oxygen. An oxygen antagonist gas is mixed with oxygen to form an oxygen gas mixture in other embodiments of the invention. Specifically contemplated, it is a mixture of oxygen gas in which the amount of oxygen in the oxygen gas mixture is less than the total amount of all gas or gases in the mixture. In some embodiments, the oxygen antagonist gas is carbon monoxide, and the amount of carbon monoxide is almost the same amount or exceeds any amount of oxygen in the oxygen gas mixture. In particular embodiments, carbon monoxide is used with biological material free of blood. The term "blood free biological matter" refers to cells and organs whose oxygenation does not depend on, or ceases to depend on, the vasculature, such as an organ for transplantation. Preferably, the atmosphere will be 100% CO, but as will be apparent to those skilled in the art, the amount of CO can be balanced with different oxygen gases, as long as the amount of usable oxygen is reduced to a level that prevents cellular respiration. . In this context, the ratio of carbon monoxide to oxygen is preferably 85:15 or higher, 199: 1 or greater or 399: 1 or greater. In certain embodiments, the ratio is approximately, at least approximately, or at most approximately 1: 1, 2: 1, 2.5: 1, 3: 1, 4: 1, 5: 1, 6: 1, 7: 1, 8: 1, 9: 1, 10: 1, 15: 1, : 1, 25: 1, 30: 1, 35: 1, 40: 1, 45: 1, 50: 1, 55: 1, 60: 1, 65: 1, 70: 1, 75: 1, 80: 1, 85: 1, 90: 1, 95: 1, 100: 1, 110: 1, 120: 1, 130: 1, 140: 1, 150: 1, 160: 1, 170: 1, 180: 1, 190: 1, 200: 1, 210: 1, 220: 1, 230: 1, 240: 1, 250: 1, 260: 1, 270: 1, 280: 1, 290: 1, 300: 1, 310: 1, 320: 1, 330: 1, 340: 1, 350: 1, 360: 1, 370: 1, 380: 1, 390: 1, 400: 1, 410: 1, 420: 1, 430: 1, 440: 1, 450: 1, 460: 1, 470: 1, 480: 1, 490: 1, 500: 1 or more, or any derivable scale in these numbers. In other embodiments, the above numbers pertain to the ratio of carbon monoxide to a mixture of oxygen and one or more other gases. In some cases, it is contemplated that the other gas is a non-reactive gas such as nitrogen (N2). Thus, in other embodiments of the invention, the foregoing numbers apply to carbon monoxide ratios to a combination of oxygen and nitrogen (O / N2) that can be used in the methods and apparatus of the invention. Accordingly, it will be understood that other gases may or may not be present. In some embodiments, the CO: oxygen ratio is balanced with one or more other gases (gases other than carbon monoxide and oxygen). In particular modalities, the CO: oxygen ratio is balanced with nitrogen. In other embodiments, the amount of CO is a ratio of CO compared to ambient air, as described by the preceding numbers. In some cases, the amount of carbon monoxide is relative to the amount of oxygen, while in others, it is an absolute amount. For example, in some embodiments of the invention, the amount of oxygen is in terms of "parts per million (ppm)", which is a measurement of the volume parts of oxygen in one million parts of air at standard temperature and pressure. of 20 ° C and pressure of one atmosphere, and the gas volume balance is made with carbon monoxide. In this context, the amount of carbon monoxide to oxygen is related in terms of parts per million of oxygen balanced with carbon monoxide. It is contemplated that the atmosphere to which the biological material is exposed or incubated may be at least 0, 50, 100, 200, 300, 400, 500 ,. 1000 or 2000 parts per million (ppm) of oxygen balanced with carbon monoxide and, in some cases, carbon monoxide mixed with a non-toxic and / or non-reactive gas. The term "environment" refers to the immediate environment of biological matter, that is, the environment with which it is in direct contact. Thus, the biological material must be directly exposed to carbon monoxide, and it is insufficient for a sealed tank of carbon monoxide to be in the same place as the biological material, and it is considered that an "environment" will be incubated in accordance with the invention. Alternatively, the atmosphere can be expressed in terms of kPa. It is generally understood that 1 million parts = 101 kPa at 1 atmosphere. In embodiments of the invention, the environment in which a biological material is incubated or exposed is approximately, at least approximately, or at most approximately 0.001, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08 , 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.30, 0.35, 0.40, 0.45 , 0.50, 0.55, 0.60, 0.65, EYE, 0.75, 0.80, 0.85, 0.90, 0.95, 1.0 kPa or more O2, or any derivable scale in these numbers. As described above, said levels can be balanced with carbon monoxide and / or other non-toxic and / or non-reactive gases. Also, the atmosphere can be defined in terms of CO levels in units of kPa. In certain embodiments, the atmosphere is approximately, at least approximately, or at most approximately 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 101, 101.3 kPa of CO, or any scale derivable in these numbers. In particular embodiments, the partial pressure is about or at least about 85, 90, 95, 101, 101.3 kPa of CO, or any scale derivable in these numbers. The amount of time the sample is incubated or exposed to carbon monoxide may also vary in the modalities of the invention.

In some embodiments, the sample is incubated or exposed to carbon monoxide by approximately, by at least approximately, or by at most approximately 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60 or more minutes and / or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, and / or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more days. The biological material is exposed to gas in a closed container in some embodiments of the invention. In some cases, the closed container can maintain a particular environment, or it can modulate the environment as desired. The environment refers to the amount of oxygen antagonist to which the biological material is exposed, and / or the temperature of the environment. In some cases, the biological material is placed under a vacuum before, during or after exposure to an oxygen antagonist. In addition, in some cases, biological matter is exposed to a normoxic environment after being exposed to an oxygen antagonist. In addition, in other embodiments, the environment containing the biological material passes through a cycle at least once at a different amount or concentration than that of the oxygen antagonist, wherein the difference in quantity or concentration is at least one difference in percentage. The environment may cycle back and forth between one or more amounts or concentrations of the oxygen antagonist, or may gradually increase or decrease the amount or concentration of an oxygen antagonist. In some cases, the different amount or concentration is between about 0 and 99.9% of the amount or concentration of the oxygen antagonist to which the biological material was initially exposed. It is contemplated that the difference in amount and / or concentration is approximately, at least approximately, or at most approximately 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 07, 0.8, 0.9, 1, 2, 3, 4, 5 , 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 , 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55 , 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 , 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any scale derivable in these numbers.

The methods of the invention may also include a step of subjecting biological matter to a temperature controlled environment. In certain embodiments, biological matter is exposed to a temperature that is a "non-physiological temperature environment," which refers to a temperature at which biological matter can not live for more than 96 hours. The temperature controlled environment may have a temperature of about, at least about, or at most about -210, -200, -190, -180, -170, -160, -150, -140, -130, -120. , -110, -100, -90, -80, -70, -60, -50, -40, -30, -20, -10, -5, 0, 1, 2, 3, 4, 5, 6 , 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56 , 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81 , 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106 , 107, - 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 1 65, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 195, 197, 198, 199, 200 ° C or more , or any differentiable scale in these numbers. The biological material can also be exposed to an oxygen antagonist at room temperature, which means a temperature between about 20 ° C and about 25 ° C. Furthermore, it is contemplated that the biological material reaches a temperature of the central part of any quantity or scale of quantities discussed. It is contemplated that the biological material may be subjected to a non-physiological temperature environment or a controlled temperature environment during or after exposure to the oxygen antagonist. In addition, in some embodiments, the biological material is subjected to a non-physiological temperature environment or a temperature controlled environment for a period between about one minute and about one year. The amount of time may be approximately, at least approximately, or at most approximately 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months , 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more years, and any combination or scale derivable in these numbers. In addition, there may also be a step of increasing the ambient temperature relative to the reduced temperature. In addition, it is contemplated that the temperature may be altered or that it may pass through a cycle during the procedure. In some embodiments, the temperature of the biological material can be reduced first before it is placed in the oxygen antagonist environment, while in others, the biological material can be cooled by placing it in the oxygen antagonist environment, which is below the temperature of biological matter. The biological material and / or the environment can be cooled or heated gradually, so that the temperature of the biological material or the environment starts at a temperature, but then reaches another temperature. In certain modalities, methods include modulating environmental oxygen levels or removing biological material from an oxygen-containing environment. Operationally, exposure of the biological material to an environment in which oxygen is diminished or absent can mimic the exposure of the biological material to an oxygen antagonist. In the methods of the invention, there is also a step of evaluating the level of the oxygen antagonist and / or the oxidative phosphorylation in the biological material in which the stasis was induced. Compositions, methods and articles of manufacture of the invention can be used in biological material that will be transferred back into the donor organism from which it was derived (autologous) or a different receptor (heterologous) subject. In some modalities, biological matter is obtained directly from a donor organism. In others, the biological matter is put into culture before exposure to an oxygen antagonist. In some situations, biological matter is obtained from a donor organism that was given extracorporeal membrane oxygenation before the recovery of biological matter, which is a technique implemented to facilitate the preservation of biological matter. In addition, the methods include the administration or implantation of the biological material in which stasis was induced, to a living receptor organism. The methods of the invention also relate to the induction of stasis in biological matter in vivo, which comprises incubating biological matter with an oxygen antagonist which creates hypoxic conditions for an effective amount of time for the biological matter to enter stasis. In addition, other embodiments of the invention include methods for reducing the oxygen demand in biological material in vivo., which comprise contacting the biological material with an effective amount of an oxygen antagonist, to reduce its oxygen demand. It is contemplated that the oxygen demands be reduced by at least about, or at most approximately, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%, or any scale derivable in these numbers, with respect to the amount of oxygen demand in cells of biological matter or a representative sample of cells of biological material not exposed or that has been left exposed to the oxygen antagonist. Other aspects of the invention relate to methods for the preservation of biological material in vivo, which comprise exposing the biological material in vivo to an effective amount of an oxygen antagonist, to preserve the biological material in vivo. The present invention also relates to a method for delaying the effects of trauma on or in an organism, which comprises exposing the biological matter at risk of trauma, to an effective amount of an oxygen antagonist. In other aspects of the invention, there are methods of treating or preventing hemorrhagic shock in a patient, comprising exposing the patient to an effective amount of an oxygen antagonist. Methods to reduce the heart rate in an organism, are They also include as part of the invention. Such methods include contacting the biological sample or organism with an effective amount of an oxygen antagonist. One embodiment of the invention relates to a method of inducing hibernation in a mammal, comprising contacting the mammal with an effective amount of an oxygen antagonist. In another embodiment, there is a method for anesthetizing an organism, which comprises exposing the biological material in which anesthesia is desired, to an effective amount of an oxygen antagonist. It is contemplated that anesthesia may be similar to local or general anesthesia. The present invention further includes methods for protecting a mammal from radiotherapy or chemotherapy, comprising contacting the mammal with an effective amount of an oxygen antagonist before or during radiation therapy or chemotherapy. With the local administration of cancer therapy, it is specifically contemplated that the oxygen antagonist may also be administered locally to the affected organ, tissue and / or cells. In further embodiments, there are methods of treating a hyperproliferative disease (e.g., cancer) in a mammal, comprising contacting the mammal with an effective amount of an oxygen antagonist, and subjecting the mammal to hyperthermia therapy. Although the methods of the invention can be applied to the preservation of organs for transplantation, other aspects of the invention relate to the recipient organism. In some embodiments, there are methods of inhibiting the rejection of an organ transplant in a mammal, which comprise providing the mammal with an effective amount of an oxygen antagonist. Regulation of temperature in organisms can be achieved by using oxygen antagonists. In some embodiments, there is a method of treating a subject with hypothermia, comprising (a) contacting the subject with an effective amount of an oxygen antagonist, and then (b) subjecting the subject to an ambient temperature above the of the subject. In other embodiments, the present invention includes a method of treating a subject with hyperthermia, comprising (a) contacting the subject with an effective amount of an oxygen antagonist. In some cases, the treatment of hyperthermia also includes. (b) subjecting the subject to an ambient temperature that is at least about 20 ° C below that of the subject. As discussed above, exposure of the subject to a non-physiological or controlled temperature environment can be used in additional modalities. In some cases, the invention relates to a method for inducing cardioplegia in a patient undergoing bypass surgery, which comprises administering to the patient an effective amount of an oxygen antagonist. It is contemplated that the administration may be local to the heart to protect it. Other aspects of the invention relate to a method for preventing hematological shock in a patient, comprising administering to the patient an effective amount of an oxygen antagonist. In addition, there are methods for promoting wound healing in an organism, which comprise administering to the organism or wound an effective amount of an oxygen antagonist. In addition, the present invention encompasses a method for preventing or treating neurodegeneration in a mammal, which comprises administering to the mammal an effective amount of an oxygen antagonist. In cases where the biological material is being protected from damage or further damage, it is contemplated that the biological material may be exposed to an oxygen antagonist within approximately, within at least approximately, or within a maximum of approximately 30 seconds. , 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more years, and any combination or scale derivable in these numbers, after the occurrence of Initial damage (trauma or injury or degeneration). Thus, in additional modalities of the invention, the methods include an initial assessment of any damage, trauma, injury, or degeneration. The methods of the invention may include the use of an apparatus or system that maintains the environment in which biological matter is placed or exposed. The invention includes an apparatus in which an oxygen antagonist is delivered, in particular as a gas. In some embodiments, the apparatus includes a container with a sample chamber for containing the biological material, wherein the container is connected to a gas supply comprising the oxygen antagonists. It is specifically contemplated that the container may be a solid container, or that it may be flexible, such as a bag. In some embodiments, the invention is an apparatus for the preservation of cells, the apparatus comprising: a container having a sample chamber with a volume of no more than 775 liters; and a first gas supply in fluid communication with the sample chamber, the first gas supply including carbon monoxide. In other embodiments, the apparatus also includes a cooling unit that regulates the temperature within the sample chamber and / or a gas regulator that regulates the amount of oxygen antagonist in the chamber or the amount of oxygen antagonist in a solution that is in the camera. It is contemplated that there may be a gas supply for a second gas or additional gas, or a second supply of gas or additional gas for the oxygen antagonist. The second gas supply may be connected to the sample chamber, or may be connected to the first gas supply. The additional gas, as discussed above, can be a non-toxic and / or non-reactive gas. A gas regulator forms part of the apparatus in some embodiments of the invention. One, two, three or more gas regulators can be used. In some cases, the gas regulator regulates the gas supplied to the sample chamber of the first gas supply. Alternatively, regulate the gas supplied to the sample chamber or first gas supply of the second gas supply, or there may be a regulator for the first and second gas supplies. It is further contemplated that any gas regulator may be programmed to control the amount of gas supplied to the sample chamber and / or to another gas supply. The regulation may or may not be for a specified period. There may be a gas regulator, which may or may not be programmable, for any gas supply directly or indirectly connected to the sample chamber. In some cases, the gas regulator is electronically programmable.

In some cases, the pressure and / or temperature inside the chamber can be regulated with a pressure regulator or temperature regulator, respectively. As with the gas regulator, these regulators can be electronically programmable. The apparatus of the invention may also have a cooling and / or heating unit to reach the temperatures discussed above. The unit may or may not be electronically programmable. In additional embodiments, the apparatus includes a wheeled cart on which the container rests, or may have one or more handles. It is specifically contemplated that the invention includes an apparatus for cells, tissues, organs and even whole organisms, in which the apparatus has: a container having a sample chamber; a first gas supply in fluid communication with the sample chamber, the first gas supply including the oxygen antagonists; and an electronically programmable gas regulator that regulates the gas supplied to the sample chamber from the first gas supply. In some embodiments, the apparatus also has a structure configured to provide a vacuum within the sample chamber. In addition, any oxygen antagonist described in this application is contemplated for use with the apparatus of the invention. In specific embodiments, carbon monoxide can be administered using this apparatus. In other cases, a chalcogenide compound or a compound having the structure of reducing agent can be administered. In addition, the present invention relates to selection tests. In some embodiments, a candidate substance is selected for its ability to act as an oxygen antagonist. This can be done using any test described herein, such as measuring the production of carbon dioxide. Any substance that has been identified has the characteristics of an oxygen antagonist, it can also be characterized or tested. Furthermore, it is contemplated that said substance may be administered to biological matter to induce stasis, or that it may be manufactured afterwards. In fact, it is understood that any method of treatment can be used in the context of a preparation of a medicament for the treatment of, or protection against, the specified disease or condition. This includes, but is not limited to, the preparation of a medication for the treatment of hemorrhagic or haematological shock, wounds and tissue damage, hyperthermia, hypothermia, neurodegeneration, sepsis, cancer and trauma. In addition, the invention includes, but is not limited to, the preparation of a medicament for a treatment to prevent shock, trauma, rejection of organs or tissues, damage by cancer therapy, neurodegeneration and injury or tissue damage. Any modality discussed with respect to one aspect of the invention also applies to other aspects of the invention.

It is understood that the embodiments in the examples section are embodiments of the invention, which are applicable to all aspects of the invention. The use of the term "or" in the claims is to indicate "and / or", unless it is explicitly stated that it refers only to alternatives, or that the alternatives are mutually exclusive, although the description supports a definition that refers only alternatives "and / or". Throughout this application, the term "approximately" is used to indicate that a value includes the standard error deviation for the device or method that is being used to determine the value. Following the durable patent law, the terms "a" and "an", when used in conjunction with the term "comprising" in the claims or in the specification, denote one or more, unless specifically indicated otherwise. way. Other objects, features and advantages of the present invention will become apparent from the following detailed description. However, it should be understood that the detailed description and specific examples, while indicating specific embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to the inventors. skilled in the art from this detailed description.

BRIEF DESCRIPTION OF THE FIGURES The following figures are part of the present specification, and are included to better demonstrate certain aspects of the present invention. The invention can be better understood by referring to one or more of these figures, in combination with the detailed description of specific embodiments presented herein. Figure 1 - Human keratinocytes survive exposure to 100% CO. Cells were visually inspected, using an inverted phase contrast microscope. Quantification of the number of viable keratinocytes estimated by staining with tripane blue, which is an indicator of cell death. Figure 2 - Discontinuity of survival in hypoxia. Viabilities were tested to adulthood, after exposure to 24 hours of anoxia (pure N2), intermediate hypoxia (0.01 kPa of O2, 0.05 kPa of 02 or 0.1 kPa of O2) or moderate hypoxia (0.5 kPa of 02) in wild type embryos. All data points are the result of at least 3 independent experiments, and the values that could not be explained, were subtracted from the total. Figure 3 - Carbon monoxide protects against hypoxia. Viabilities were tested to adulthood, after exposure to 24 hours of pure carbon monoxide, 0.05 kPa O2 / N or 0.05 kPa of 02 / CO in wild-type embryos. All data points are the result of at least 3 independent experiments, and the values that could not be explained were subtracted from the total. Figure 4A - The metabolic rate decreases before the temperature of the central part of the body is reached when the mice are exposed to hydrogen sulfide. The exposure of mice at 80 ppm (at 0 minutes on the X axis), results in an approximately 3-fold decrease in CO2 production (black line) in less than five minutes. This precedes the decrease in temperature of the central part of the animal to the ambient temperature (gray line). Figure 4B - Temperature of mice exposed to hydrogen sulfide. Each trace represents a continuous measurement of the temperature of the central part of the body of individual mice exposed to 80 ppm of H2S, or to ambient air. The numbers on the vertical axis are temperatures in degrees Celsius. On the horizontal axis, the numbers reflect the time in hours. The experiments were carried out for 6 hours, followed by recovery records. The starting point is at 1:00, and at the end of the 6-hour treatment, it is approximately 7:00. Figure 5A-5C - Exposure to 80 ppm of hydrogen sulfide causes the temperature of the central part of a mouse body to approach room temperature. The gas tap was opened and the temperature was lowered starting at 0:00. The atmosphere was changed back to ambient air at 6:00. The triangles indicate the temperature of the central part of the mouse body, determined by radiotelemetry. This was approximately 39 ° C at 0:00. The diamonds indicate the ambient temperature that was reduced from 23 ° C to 13 ° C in the first 3 hours of the experiment, and then increased again to 23 ° C from 6:00 o'clock, stabilizing around 9:00 o'clock. Figure 5A shows the change in temperature of the central part of the body as a result of hydrogen sulfide. Figure 5B is an enlarged version of Figure 5A. Figure 5C shows the control animals. Figure 6 - The rate of decrease of the temperature of the central part of the body, depends on the concentration of hydrogen sulfide administered to the mice. All the lines represent the temperature of the central part of the body of an individual mouse determined by radiotelemetry. Mice subjected to 20 ppm and 40 ppm H2S exhibit minor decreases in core body temperature. Exposure to 60 ppm induced a substantial decrease in temperature, starting at approximately 4:00. The mouse exposed to 80 ppm exhibited a substantial decrease in temperature, starting at approximately 2:00. Figure 7 - Lower temperature of the central part of the body. The lowest temperature of the central part of the body recorded for a mouse exposed to 80 ppm of hydrogen sulfide was 10J ° C. The triangles indicate the temperature of the central part of the mouse body determined by radiotelemetry, which was started at approximately 39 ° C at time 0. The diamonds indicate the ambient temperature that started at approximately 23 ° C, and decreased to less than 10 ° C. ° C around the midpoint of the experiment, after which it then increased again to room temperature. Figure 8 A - Endogenous levels of hydrogen sulfide are increased in mice acclimatized at warm temperatures. The gray bars (two bars on the left) indicate endogenous H2S concentrations of two individual mice acclimated at 4 ° C; the black bars (two bars on the right) indicate the endogenous H2S concentrations of two individual mice acclimated at 30 ° C. Concentration of hydrogen sulfide determined by GC / MS. Figure 8B - Effects of ambient temperature on the temperature decrease dependent on hydrogen sulfide. The rate of decrease of the temperature of the central part of the body (expressed in degrees centigrade) due to exposure to hydrogen sulfide, depends on the acclimatization temperature. The mice were exposed to the gas at 1:00 am. The triangles indicate the temperature of the central part of the mouse body, acclimated at 12 ° C, determined by radiotelemetry. The squares indicate the temperature of the central part of the body of the animal acclimated at 30 ° C. Figure 9 is a block diagram illustrating a breathing gas supply system in accordance with embodiments of the present invention.

Figure 10 is a schematic drawing illustrating a breathing gas supply system in accordance with embodiments of the present invention. Figure 11 is a schematic drawing illustrating a breathing gas supply system in accordance with other embodiments of the present invention. Fig. 12 is a flow chart illustrating operations in accordance with embodiments of the present invention. Fig. 13 is a schematic drawing illustrating a gas supply system for treating fabrics in accordance with embodiments of the present invention. Figure 14 is a flow chart illustrating operations in accordance with embodiments of the present invention. Figure 15 - Metabolic inhibition protects against death induced by hypothermia in nematodes. Nematodes exposed to low temperatures (4 ° C), are unable to survive after 24 hours. However, if they are kept in anoxic conditions during the hypothermia period (and for a period of 1 hour before and after), a substantial proportion of the nematodes survive.

DETAILED DESCRIPTION OF THE ILLUSTRATIVE MODALITIES Stasis In "stasis" or "suspended animation", a cell, tissue or organ or organism (referred to collectively as "biological material") is alive, but cellular functions necessary for cell division, progression of development and metabolic status are delayed or even stopped. This state is desirable in many contexts. Stasis can be used as a preservation method on its own, or it can be induced as part of a cryopreservation regimen. Biological materials can be preserved for research use, for transportation, for transplantation, for therapeutic treatment (such as ex vivo therapy), and to prevent the onset of trauma, for example. Stasis with respect to whole organisms has similar uses. For example, the transportation of organisms could be facilitated if they had entered into stasis. This could reduce the physical and physiological damage to the body, reducing or eliminating stress or physical injury. These modalities are discussed in more detail below. Stasis can be beneficial by decreasing the need for biological material by oxygen and, therefore, blood flow. It can extend the period that biological material can be isolated from an environment that sustains life and can be exposed to an environment that induces death. Although recovery from accidental hypothermia has been reported for a relatively long period (Gilbert et al., 2000), there has been recent interest in intentionally inducing suspended animation in organisms (the discussion of any reference will not be considered as an admission that the reference In fact, some references discussed in the present would not be prior art with respect to priority applications). Controlled hyperthermia has been explored, as well as the administration of a cold jet of a solution in the aorta (Tisherman, 2004), induction of cardiac arrest (Behringer et al., 2003) or suspended animation induced by nitric oxide (Teodoro et al. ., 2004). An organism in stasis is distinguishable from an organism under general anesthesia. For example, an organism in moderate stasis (decrease in cellular respiration between approximately 2 and approximately 5 times) that is exposed to ambient air will begin to shiver, while an organism under anesthesia will not. Likewise, it is anticipated that an organism in moderate stasis responds to the squeeze of a toe, while an organism under anesthesia usually does not. Therefore, stasis is not the same as being under anesthesia, as is commonly practiced. The present invention is based on the observation that certain types of compounds effectively induce reversible stasis in biological matter.

A. Thermoregulation Stasis in a warm-blooded animal will affect thermoregulation. Thermoregulation is a characteristic of so-called "warm blood" animals, which allows the body to maintain a relatively constant core temperature when exposed to significantly altered environmental temperatures (cold or heat). The ability to control thermoregulation by stasis induction is an aspect of the invention, and allows uses similar to those discussed above. The thermal regulation can be facilitated by placing organisms, extremities or organs or tissues isolated in cameras / devices, whose temperature can be controlled. For example, hot chamber-like devices or devices similar to hyperbaric chambers can encompass an entire organism, and can be connected to thermoregulatory devices. Smaller devices such as blankets, sleeves, cuffs or gloves are also contemplated (e.g., CORE CONTROL cooling system, by AVAcore Technologies, Palo Alto, CA, U.S. Patent No. 6,602,277). Said devices / cameras can be used to increase or decrease the ambient temperatures.

B. Biological material The biological material contemplated for use with the present invention includes material derived from invertebrates and vertebrates, including mammals; biological materials include organisms. In addition to humans, the invention may be used with respect to mammals of veterinary or agricultural importance, including those of the following classes: canine, feline, equine, bovine, ovine, murine, porcine, caprine, rodent, lagomorph, lupine and ursine. . The invention also extends to fish and birds. Other examples are described below. In addition, the type of biological matter varies. It can be cells, tissues and organs, as well as organisms for which different compositions, methods and devices have relevance. The patent applications of E.U.A. not provisional, titled "Methods, Compositions and Devices for Inducing Stasis in Cells" and "Methods, Compositions and Devices for Inducing Stasis in Tissues and Organs", in the name of Mark. B. Roth, filed on October 22, 2004, are hereby incorporated by reference in their entirety. 1. Different sources The following are examples of sources from which biological matter can be obtained. The embodiments of the invention include, but are not limited to, these examples. to. Mammals In certain aspects of the invention, the mammal is of the Order Monotremata, Marsupiaüa, Insectívora, Macroscelldia, Dermoptera, Chiroptera, Scandentia, Primates, Xenarthra, Pholidota, Tubulidentata, Lagomorpha, Rodentia, Cetacea, Carnivora, Proboscidea, Hyracoidea, Sirenia, Perissodactyla or Artiodactyla. Examples of the Monotremata Order include the Tachyglossidae families (eg, echidnas) and Omithorhynchidae (eg, Platypus). Examples of the Marsupialia Order include the Didelphidae (for example, possums), Microbiotheriidae (for example, mountain monkey), Caenolestidae (for example, rat possums), Dasyuridae (for example, marsupial mouse), Myrmecobiidae (for example, numbat) families. ), Thylacinidae (for example, Thylacine), Peramelidae (for example, bandicoots), Thylacomyidae (for example, rabbit bandicuts), Notoryctidae (for example, marsupial moles), Phalangeridae (for example, couscous), Petauridae (for example, cacomiztles) , gliders), Burramyidae (for example, pygmy possums), Macropodidae (for example, kangaroos, wallabies), Tarsipedidae (for example, Tarsipes spencerae), Vombatidae (for example, wombats) and Phascolarctidae (for example, koalas). Insectivorous Order includes, for example, families Solenodontidae (eg, airs), Tenrecidae (eg, tenrecs, shrews otter), Chrysochloridae (eg, golden moles), Erinaceidae (eg, hedgehogs, Echin-osorex gymnurus), Soricidae (eg, shrews) and Talpidae (for example, moles, outrages). The Macroscelidia Order includes the Macroscelidia family (for example, elephant shrews). The Scandentia Order includes the family Tupaiidae (for example, arboreal shrews). The Dermoptera Order includes the Cynocephalidea family (eg, flying lemurs). The Chiroptera Order includes the families Pteropodidae (eg, fruit bats, flying foxes), Rinopomatidae (eg, mouse tailed bats), Craseonycteridae (eg, bat-nosed bat or bumblebee bat), Emballonuridae (eg, Pod-tail bats), Nycteridae (eg, slot-face bats), Megadermatidae (eg, false vampire bats), Rhinolophidae (eg, horseshoe bats), Noctilionidae (eg, buldog bats, fishing bats), Mormoopidae , Phyllostomidae (for example, New World leaf nose bats), Natalidae, Furipteridae, Thyropteridae, Myzapodidae, Vespertilionidae (for example, common bats), Mystacinidae (for example, short-tailed bats) and Molossjdae (for example, bats). free tail). The Primates Order includes the families Lemuridae (for example, lemurs), Cheirogaleidae (for example, mouse lemurs), Indriidae (for example, indri, woolly lemur), Daubentoniidae (for example, aye-aye), Lorisidae (for example, lorises , Galago, gálagos), Tarsiidae (for example, tarseros), Cebidae (for example, New World monkeys, marmosets, tamarins), Hylobatidae (for example, gibbons), Pongidae (for example, apes) and Hominidae (for example, man). Examples of the Xenarthra Order include families Myrmecophagidae (for example, tamandúas), Bradypodidae (for example, three-toed sloths), Megalonychidae (for example, two-toed sloths) and Dasypodidae (for example, armadillos). Examples of the Pholidota Order include the Manidae family (eg, pangolins). Examples of the Tubulidentata Order include the Orycteropodidae family (eg, swine). Examples of the Order Lagomorpha include the families Ochotonidae (for example, pikas) and Leporidae (for example, hares and rabbits). The Rodentia Order includes the families Aplodontidae (for example, mountain beavers), Scluridae (for example, squirrels, marmots, chipmunks), Geomyidae (for example, pocket terrestrial squirrels), Heteromyidae (for example, pocket mice, kangaroo rats), Castoridae (for example, beaver), Anomaluridae (for example, scaly-tailed squirrels), Pedetidae (for example, jumping hare), Muridae (for example, rats and mice), Gliridae (for example, dormice), Selevinüdae (for example, desert dormouse), Zapodidae (for example, jumping mice), Dipodidae (for example, gerbils), Hystricidae (for example, Old World porcupines), Erethizontidae (for example, New World porcupines) , Caviidae (for example, guinea pigs, maras), Hydrochaeridae (for example, capibara), Dinomyidae (for example, false paca), Agoutidae (for example, pacas), Dasyproctidae (for example, agouti), Chinchillidae (for example, , chinchillas, vizcachas), Capromyidae (por example, hutias), Myocastoridae (eg, otter), Ctenomyidae (eg, tuco-tucos), Octodontidae (e.g., octodon, Octodon), Abrocomidae (e.g., chinchilla rats), Echimyidae (e.g., spiny rats) , Thryonomyidae (for example, cane rats), Petromyidae (for example, rat of African rocks), Bathyergidae (for example, mole rat) and Ctenodactylidae (for example, gundis).

The Cetacean Order includes the Iniidae families (for example, Amazon porpoise), Lipotidae, Platanistidae, Pontoporiidae, Ziphiidae (for example, bill whales), Physeteridae (for example, sperm whales), Monodontidae (for example, beluga, narwhal), Delphinidae (for example, marine dolphins, oreas), Phocoenidae (for example, porpoises), Balaenopteridae (for example, rorquals), Balaenidae (for example, true whales) and Eschrichtiidae (for example, gray whales). Carnivorous Order includes Canidae families (eg, dogs, foxes, wolves, jackals, coyotes), Ursidae (eg, bears), Procyonidae (eg, raccoons, coatis, kinkayus, lesser pandas), Ailuropodidae (eg, giant pandas), Mustelidae (for example, weasels, skunks, badgers, otters), Viverridae (for example, civets, jinetas), Herpestidae (for example, mongooses), Protelidae (for example, South African hyena), Hyaenidae (for example, hyenas), Felidae (for example, cats), Otariidae (for example, ear seals, sea lions), Odobenidae (for example, walruses) and Phocidae (for example, seals without ears). The Proboscidea Order includes the Elephantidae family (for example, elephants). The Hyracoidea Order includes the family Procaviidae (for example, hiráceos). The Sirenia Order includes the families Dugongidae (for example, dugong) and Trichechidae (for example, manatees). The Perissodactyla Order includes the Equidae families (for example, horses, donkeys, zebras), Tapiridae (for example, tapirs) and Rhinocerotidae (for example, rhinoceroses). The Artiodactyla Order includes the Suidae families (eg, pigs, babirusa), Tayassuidae (eg, peccaries), Hippopotamidae (eg, hippos), Camelidae (eg, camels, llamas, vicuñas), Tragulidae (eg, chevrotaines) ), Moschidae (for example, musk deer), Cervidae (for example, deer, elk, suede), Giraffidae (for example, giraffe, okapi), Antilocapridae (for example, pronghorn) and Bovidae (for example, cattle, sheep, antelopes, goats). b. Reptiles In certain modalities, the biological material is a reptile, or it is derived from a reptile. The reptile can be of the Order Chelonia, Pleurodira, Squamata, Rhynchocephalia or Crocodylia. A reptile of the Order Chelonia can be, for example, a member of the families Carettochelyidae, Chelydridae (for example, biting turtles), Cheloniidae (for example, biting turtles, green turtles), Dermatemydidae (for example, lutes), Emydidae (for example, painted turtles, pond sliders, pond turtles, snail-eating turtles, chest turtles), Kinostemidae (eg musk turtles), Saurotypidae, Testudinidae (eg, Galápagos tortoises, desert tortoises, turtles of Aldabra, spu-thighed turtles, Hermann's turtles), Trionychidae (for example, soft shell turtles of China, spiny soft-shell turtles) or Platystemidae. A reptile of the Pleurodira Order can be, for example, a member of the Chelidae families (for example, snake-neck turtles) or Pelomedusidae (for example, armored turtles).

A reptile of the Squamata Order may be, for example, a member of the Agamidae families (for example, rainbow lizards, bearded dragons, leeches from India, spiny-tailed lizards), Chamaeleontdidae (for example, chameleons), Iguanidae (e.g. , anolis, basilisks, collar lizards, guanas, horned lizards, chacahualas, mugwort lizards, side-blotched lizards), Gekkonidae (for example, geckos), Pygopodidae, Teüdae (for example, lizards listed, Tupinambis), Lacertidae (for example, sand lizards, ocellated lizards, viviparous lizards, wall lizards, long-tailed lizards), Xantuslidae, Scincidae (for example, estincos), Cordylidae (for example, sungazers), Dibamidae, Xenosauridae, Anguidae (e.g. , lution, alligator lizards, sheltopusick, crystal lizards), Helodermatidae (for example, Gila monster), Lanthanotidae, Varanidae (for example, monitors), Leptotyphlopidae, Typhlopidae, Anomalepididae, Anili idae (eg, tube snakes), Uropeitidae, Xenopeltidae, Boidae (eg, boas, anacondas, - pitones from rocks), Acrochordidae (eg, warty snakes), Colubridae (eg, mangrove snakes, whip snakes) , smooth snakes, egg-eating snakes, boomslangs, rat snakes ,. Aesculapius snakes, four-line serpents, oriental snake, tentacled serpents, heterodons, king snakes, Montpelier snakes, grass serpents, water snakes, snakelike snakes, twig snakes, backbone snakes), Elapidae (for example, Australian vipers, kraits, mambas, coralillos, cobras, copperhead snakes, desert vipers), Viperidae (for example, vipers, true common vipers, rattlesnakes, rattlesnakes, snakes), Hydrophiidae (eg, sea brait), Amphisbaenidae (for example, lizard worm, Bipedidae or Trogonophidae (for example, lizard of the burrows) A reptile of the Order Rhynchocephalia can, for example, a member of the family Sphenodontidae (for example, tuataras) .A reptile of the Order Crocodylia can be, for example, a member of the families Alllgatoridae (eg, lizards, alligator), Crocodylidae (eg, crocodiles) or Gavialidae (eg, gaviales). c. Amphibians The biological material of the present invention can be an amphibian, or it can be derived from an amphibian. Amphibians can be, for example, a frog or a frog. The frog or frog may be, for example, a member of the families Anthroleptidae (e.g., chilling frogs), Ascaphidae (e.g., frogs with tails), Brachycephalldae (e.g., golden frogs and shield toads), Bufonidae ( for example, true toads), Centrolenidae (eg, glass frogs and leaf frogs), Dendrobatidae (eg poison dart frogs), Discoglossidae (eg, fire-bellied toads), Heleophrynidae (eg, ghost frogs) , Hemisotidae (for example, shovel nose frogs), Hylidae (for example, New World tree frogs), Hyperoliidae (for example, African tree frogs), Leiopelmatidae (for example, frogs from New Zealand), Leptodactylidae (for example, Neotropical frogs), Megophryidae (e.g., frogs from South Asia), Microhylidae (e.g., microhy- droid frogs), Myobatrachidae (e.g., Australian frogs), Pelobatidae (e.g., shovel-toed toads), Pelodytidae (e.g. frogs parsley), Pipidae (for example, mute frogs), Pseudidae (for example, paradox frogs), Ranidae (for example, river frogs and true frogs), Rhacophoridae (for example, Old World tree frogs), Rhinodermatidae (for example, Darwln frogs) , Rhinophrynidae (for example, toads from burrows), Sooglossidae (for example, frogs from Seychelle), Caudata (for example, salamanders) or Gymnophiona (for example, Cecilias). The amphibian can be a salamander. The salamander can be, for example, a member of the Ambystomatidae families (for example, mole salamanders), Amphiumidae (eg, Congo salamander), Cryptobranchidae (eg, giant salamanders and giant water salamanders), Dicamptodontidae (eg, giant salamanders of the Pacific), Hynobiidae (eg, Asian salamanders), Plethodontidae ( for example, non-lung salamanders), Proteidae (for example, axolotls and water dogs), Rhyacotritonidae (for example, salamanders of the torrents), Salamandridae (for example, tritons and salamanders) or Sirenidae (for example, sirens). Alternatively, the amphibian may be a member of the Caecilian family. The member of the Caecilian family can be, for example, a member of the families Caeciliidae (for example, Cecilias), Ichthyophiidae (for example, Asian Cecilia tail), Rhinatrematidae (for example, Cecilia de cola neotropicales), Scolecomorphidae (por example, African Cecilia), Typhlonectidae (eg, Cecilia aquatica) or Uraeotyphlidae (eg Cecilia from India). d. Birds The biological material of the present invention can be a bird, or it can be derived from a bird. The bird may be, for example, a member of the Order Anseriforme (for example, aquatic bird), Apodiforme (for example, hummingbirds and swifts), Cap imulgiforme (for example, nocturnal birds), Charadriiforme (for example, birds of the coast ), Ciconiiforme (for example, storks), Coliiforme (for example, collús), Columbiforme (for example, pigeons and pigeons), Coraciiforme (for example, kingfisher), Craciforme (for example, chachalacas, guacos, guans, megápodos) , Cuculiforme (for example, cuckoos, hoactzin, turacos), Falconiformes (for example, diurnal birds of prey), Galliforme (for example, chicken-like birds), Gavüforme (for example, loons), Guiforme (for example, negretas, cranes , rails), Passeriformes (for example, percher rats), Pelecaniformes (for example, pelicans), Phoenicopteriformes (for example, flamingos), Piciformes (for example, woodpeckers), Podicipediforme (for example, loons), Procellariiforme (for example, , pipe-nosed birds), Psi ttaciform (eg, parrots), Sphenisciform (eg, penguins), Strigiform (eg, owls), Struthioniforme (eg cassowaries, emus, ki is, ostriches, rheas), Tinamiforme (eg, tinamou), Trogoniforme (for example, quetzales) or Turniciforme (for example, button quail). and. Fish The biological material of the present invention can be a fish, or it can be derived from a fish. The fish can be, for example, a member of the Order Acipenseriforme (for example, leaffish, spoonfish and sturgeon), Polypteriformes (for example, bichiers, birchers, lobe-fingered pike and reed fish), Atheriniforme (for example, fish rainbow and silvery-flanked fish), Beloniform (eg, half-spiked fish and needle fish), Beryciform, Channiform, Cyprinodontiforme (eg, gobies), Dactylopteriforme (eg, flying fish), Gasterosteiforme (eg. sea and spiny fish), Mugiliforme (for example, smooth), Pegasiforme (for example, dragon fish and sea dragons), Perciform (for example, perch-type fish), Pleuronectiforme (for example, flounders, flounders and soles), Scorpaeniforme (for example, scorpion fish and rockfish), Stephanoberyciform, Synbranchiforme (eg, swamp eels), Tetraodontiforme (eg, boxfishes, triggerfish, Oligoplites, puffer fish, triggerfish and chapines), Zeiforme (e.g. example, wild boar fish, rooster fish and ceos), Atherinomorpha, Clupeiforme (eg, anchovetas and herring), Aulopiforme, Albuliforme, Anguilliforme (eg, eels), Elopiforme (eg, tarpon), Notacanthiformes (eg, spiny eels) and tapir fish), Saccopharyngiformes, Lampridiforme (for example, moonfish and sea tails), Characiform (for example, clefts and piranhas), Cypriniforme (for example, cyprinids, catostomids, zebrafish), Gonorhynchiformes (for example, Chanos chanos and shellears), Gymnotiform, Siluriform (eg, catfish), Aphredoderiforme (eg, cave fish and pirate perch), Batrachoidiforme, Gadiforme (eg, cod and hake), Gobiesociforme, Lophiiforme (eg, pejesapos) , Ophidiiforme, Percopsiforme (for example, trouts and perches), Polymixiiforme (for example, bearded fish), Cetomimlforme, Ctenothrissiforme, Esociforme (for example, carps of the mud and pikes), Osmeriforme (for example, Argentine) s and espértanos), Salmoniformes (for example, salmon), Myctophiforme (for example, fish of Latern), Ateleopodiforme, Stomiiforme, Amiiforme (for example, amias), Semyonotiforme (for example, needles of sea), Syngnathiforme (for example, needles) of sea and littoral fish), Ceratodontiforme (eg, Australian Lungfish), Lepidosirenform (eg, Lungfish from South Africa and Lungfish from Africa) or Coelacanthiforme (eg, celacanths).

F. Invertebrates The biological material can be an invertebrate, or it can be derived from an invertebrate. The invertebrate can be, for example, a member of the Phylum Porifera (for example, sponges), Cnidaria (for example, jellyfish, hydras, sea anemones, Portuguese frigate and corals), Platyhelminthes (for example, worms that include planada, trematodes and tapeworms), Nematoda (for example, round worms, which include rotifers and nematodes), Mollusca (for example, mollusks, snails, slugs, octopus, squids), Annelida (for example, segmented worms, which include earthworms, leeches and marine worms), Echinodermata (eg, starfish, sea cucumbers, flat sea urchins, sea urchins), Phoronida (eg, horseshoe worms), Tardigrada (eg, water bears), Acanthocephala (eg example, thorny-headed worms), Ctenophora (for example, sea combs) or Arthropoda (for example, arachnids, crustaceans, millipedes (millipedes), centipedes (centipedes), insects). An arthropod can be, for example, a member of the following Orders and Classes: Coleoptera (for example, beetles), Diptera (for example, true flies), Hymenoptera (for example, ants, bees, wasps), Lepidoptera (e.g. , butterflies, moths), Mecoptera (for example, scorpion flies), Megaloptera, Neuroptera (for example, ensopas and relatives), Siphonaptera (for example, fleas), Strepsiptera (for example, parasitic insects and parasites of braided wings), Trichoptera (for example, fríganos); Anoplura (for example, sucking lice), Hemiptera (for example, true bugs and their close friends), Mallophaga (for example, biting lice), Psocoptera (for example, psocids), Thysanoptera (for example, thrips), Orthoptera (for example, , grasshoppers, locusts), Dermaptera (for example, earwigs), Dictyoptera, Embioptera (for example, weavers of nets), Grylloblattodea, Mantophasmatodea (for example, gladiators), Plecoptera (for example, plecópteros), Zoraptera (for example, zorápteros ), Ephemeroptera (for example, Mayflies), Odonata (for example, damselflies and dragonflies or damselflies), Phasmatoptera (for example, stick insects), Thysanura (for example, silverfish), Archaeognatha, Collembola (for example, snow flies and spinytails), Chilopoda (for example, centipedes), Diplopoda (for example, millipedes), Pauropoda (for example, pauropods and progonatos), Symphyla (for example, pseudo-centipedes and synfites), Malacostraca (for example, crabs , kr III, scale insects, shrimp), Maxillopoda, Branchiopoda (for example, branchiopods), Cephalocarida, Ostracoda (for example, ostracods), Remipedia, Branchiura, Cirripedia (for example, barnacles), Arachnida (for example, arachnids, which they include ambligidids, spiders, typists, reapers, microscorps, scorpions from books, false scorpions, pseudoscorpions, scorpions, solpúgidos, sun and uropigid spiders), Merostomata (for example, bayonet crabs) or Pycnogonida (for example, sea spiders). g. Fungi The biological material of the present invention can be a fungus, or it can be derived from a fungus. The fungus may be, for example, a member of the class Ascomycota (sack fungi), Basidiomycota (hat fungi), Chytridiomycota (chytridios), Deuteromycota or Zygomycota. The fungus can be of the genera Rhizopus, Pilobolus, Arthrobotrys, Aspergillus, Allomyces, Chytridium, Agaricus, Amanita, Cortinarius, Neurospora, Morchella, Saccharomyces, Pichia, Candida and Schizosaccharomyces, or ergot of rye. In particular embodiments, the fungus may be of the species Saccharomyces cerevisiae, Schizosaccharomyces pombe, Candida albicans or Pichia pastoris. h. Plants The biological material of the present invention can be a plant, or it can be derived from a plant. The plant can be a bryophyte (for example, mosses, liverworts, ceratophiles), licophytes (for example, lycopods, pinillos), sphenophytes (for example, horsetails), pteridophyta (for example, ferns), cicadofites (for example, cicadas), gnenophyta (for example, Gnetum, Ephedra, Welwitschia), coniferophyta (for example, conifers), ginkgophyta (for example, ginkgo) or anthophyte (for example, flowering plants). The anthophyte can be a monocot or a dicot. Non-limiting examples of monocotyledonous plants include wheat, corn, rye, rice, grass for turf, sorghum, millet, sugarcane, lily, iris, agave, Aloe, orchids, bromeliads and palms. Non-limiting examples of dicotyledonous plants include tobacco, tomato, potato, soybean, sunflower, alfalfa, cañola, rose, Arabidopsis, coffee, citrus fruits, beans, alfalfa and cotton. i. Protists The biological material of the present invention can be a protist, or it can be derived from a protist. The protist may be a rhodophyte (e.g., red algae), pheophyte (e.g., brown algae, kelp), chlorophytes (e.g., green algae), eugleophytes (e.g., euglenoids), myxomycete (e.g., mucilaginous molds). , oomycete (for example, water molds, powdery mildew, potato blight) or bacillaryophyte (for example, diatoms). j. Prokaryotes In certain aspects of the invention, the biological material is a prokaryote, or is derived from a prokaryote. In certain modalities, the prokaryote is from the Archaea family (archaebacteria). Archaebacteria may be, for example, of the Crenarchaeota, Euryarchaeota, Korarchaeota or Nanoarchaeota class. In certain aspects, the member of the class Euryarchaeota is a member of the groups of: Halobacteria, Methanobacteria, Methanococci, Methanomicrobia, Methanosarcinae, Methanopyri, Archeoglobi, Thermoplasmata or Thermococci. Specific non-limiting examples of archaebacteria include: Aeropyrum pernix, Methanococcus jannaschii, Halobacterium marismortui and Thermoplasma acidophilum. In certain modalities, the prokaryote is a eubacterium. Eubacteria can be, for example, a member of the groups of: Actinobacteria, Aquificae, Bacteroidetes, green sulfur bacteria, Chlamydiae, Verrucomicrobia, Chloroflexi, Chrysiogenetes, Cyanobacteria, Deferribacters, Deinococcus-Thermus, Dictyoglomi, Fibrobacters / Acidobacteria, Firmicutes, Fusobacteria, Gemmatimonadetes, Nitrospirae, Omnibacteria, Planctomycetes, Proteobacteria, Spirochaetes, Thermodesulfobacteria or Thermotogae. Non-limiting examples of actinobacteria include bacteria of the genera Actinomyces, Arthrobacter, Corynebacterium, Frankia, Micrococcus, Micromonospora, Mycobacterium, Propionibacterium and Streptomyces. Specific examples of actinobacteria include Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium avium, Corynebacterium glutamicum, Propionibacterium acne and Rhodococcus equi. Non-limiting examples of Aquificae include bacteria from the genera Aquifex, Hydrogenivirga, Hydrogenobacter, Hydrogenobaculum, Thermocrinis, Hydrogenothermus, Persephonella, Sulfurihydrogenibium, Balnearium, Desulfurobacterium and Thermovibrio. Non-limiting examples of Firmicutes include bacteria of the genera Bacillus, Clostridium and Molecutes. Specific examples of Firmicutes include: Listeria innocua, Listeria monocytogenes, Bacillus subtilis, Bacillus anthracis, Bacillus thuringiensis, Staphylococcus aureus, Clostridium acetobutylicum, Clostridium difficile, Clostridium perfringens, Mycoplasma genitalium, Mycoplasma pneumoniae, Mycoplasma pulmonis, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus mutans , Lactococcus lactis and Enterococcus faecalis. Non-limiting examples of Chlamydlae / Verrucomicrobia, include bacteria such as Chlamydia trachomatis, Chlamydia pneumoniae and Chlamydia psittaci. Non-limiting examples of Deinococcus-Thermus include bacteria of the genera Deinococcus and Thermus.

Proteobacteria are Gram-negative bacteria. Non-limiting examples of proteobacteria include bacteria of the genera Escherichia, Salmonella, Rickettsia, Agrobacterium, Brucella, Rhizobium, Neisseria, Bordetella, Burkholderi, Buchnera, Yersinia, Klebsiella, Proteus, Shigella, Haemophilus, Pasteurella, Actinobacillus, Legionella, Mannheimia, Coxiella , Aeromonas, Francisella, Moraxella, Pseudomonas, Campylobacter and Helicobacter. Specific examples of proteobacteria include: Rickettsia conorii, Rickettsia prowazekii, Rickettsia typhi, Ehrlichia Boris, Agrobacterium tumefaciens, Brucella melitensis, Rhizobium rhizogenes, Neisseria meningitides, Bordetella parapertussis, Bordetella pertussis, Burkholderi mallei, Burkholderi pseudomallei, Neisseria gonorrhoeae, Escherichia coli, Salmonella enterica , Salmonella typhimurium, Yersinia pestis, Klebsiella pneumoniae, Yersinia enterocolitica, Proteus vulgaris, Shigella flexneri, Shigella sonnei, Shigella dysenterica, Haemophilus influenzae, Pasteurella multocida, Actinobacillus actinomycetemcomitans, Actinobacillus pleuropneumoniae, Haemophilus somnus, Legionella pneumophila, Mannheimia haemolytica, Vibrio cholerae, Vibrio parahaemolyticus, Coxiella burnetii, Aeromonas hydrophila, Aeromonas salmonicida, Francisella tularesis, Moraxella catarrhalis, Pseudomonas aeruginosa, Pseudomonas putida, Campylobacter jejuny and Helicobacter pylori. Non-limiting examples of spirochetes include bacteria from the families Brachyspiraceae, Leptospiraceae and Spirochaetaceae. Specific examples of spirochetes include Borrelia burgdri and Treponema pallidum. 2. Different types of biological matter The methods and apparatus of the invention can be applied to organisms. Stasis of the organism can be induced, or stasis can be induced within cells, tissues and / or organs of the organism. The biological material in which stasis can be induced and which is contemplated for use with the methods and apparatuses of the invention, is limited only in that it comprises cells that use oxygen to produce energy. Stasis can be induced in cells, tissues or organs that include the heart, lung, kidney, liver, bone marrow, pancreas, skin, bone, vein, artery, cornea, blood, small intestine, large intestine, brain, spinal cord, smooth muscle , skeletal muscle, ovary, testicle, uterus and umbilical cord. In addition, stasis can be induced in cells of the following type: platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, skeletal muscle cell, endocrine cell, glial cell, neuron, secretory cell, cell with function as barrier, contractile cell, absorption cell, mucosal cell, limb (cornea) cell, stem cell (totipotent, pluripotent or multipotent), fertilized or unfertilized oocyte, or sperm. In addition, stasis can be induced in plants or parts of plants, which include fruit, flowers, leaves, stems, seeds and cuttings. The plants can be agricultural, medicinal or decorative. The induction of stasis in plants can intensify the storage path or the resistance of the whole plant or part of it to pathogens. The methods and apparatus of the invention can be used to induce stasis in biological matter in vivo. This can serve to protect and / or preserve the biological material of the organism itself, or to prevent damage or injury (or more damage or injury) to them, or the organism in general. 3. Tests Stasis can be measured by many forms including quantifying the amount of oxygen consumed by a biological sample, the amount of carbon dioxide produced by the sample (indirect measurement of cellular respiration), or characterizing mobility. To determine the rate of oxygen consumption or the rate of production of carbon dioxide, the biological material is put in a chamber that is sealed with two openings; for gas entry and exit. Gas (ambient air or other gases) is passed into the chamber at a given flow rate and out of the exit orifice, to maintain approximately 1 atmosphere of pressure in the chamber. Before and after exposure to the chamber, the gas is passed through a carbon dioxide detector and / or an oxygen detector to measure (every second) the amount of each compound in the gas mixture. The comparison of these values with time gives the regime of oxygen consumption or production of carbon dioxide.

II. Oxygen Antagonists Oxygen metabolism is a fundamental requirement for life in aerobic metazoans. Aerobic respiration accounts for the vast majority of energy production in most animals, and also serves to maintain the redox potential necessary to carry out important cellular reactions. In hypoxia, decreased oxygen availability results in inefficient transfer of electrons to molecular oxygen in the final step of the electron transport chain. This inefficiency results in a decrease in the production of aerobic energy, and an increase in the production of harmful free radicals, mainly due to the premature release of electrons in the III complex and the formation of 02"by cytochrome oxidase (Semenza, 1999 Limited energy supplies and damage by free radicals can interfere with essential cellular processes such as protein synthesis and maintenance of membrane polarity (Hochachka et al., 1996), and will eventualead to cell death. .

A. Carbon monoxide Carbon monoxide (CO) is a colorless, odorless and tasteless gas that can be toxic to animals, including humans. According to the Center for Disease Control, more than 450 people die unintentionafrom carbon monoxide every year. It can be toxic to organisms whose blood has oxygen that sustains its survival. It can be poisonous by entering the lungs through normal breathing, and by displacing oxygen from the bloodstream. The interruption of the normal supply of oxygen endangers the functions of the heart, brain and other vital functions of the body. However, the use of carbon monoxide for medical applications is being explored (Ryter et al., 2004). At amounts of 50 parts per million (ppm), carbon monoxide does not present symptoms for humans exposed to it. However, at 200 ppm, within two to three hours, carbon monoxide can cause a slight headache; at 400 ppm, within one to two hours, it can cause a frontal headache that can become widespread within three hours; and at 800 ppm, it can cause dizziness, nausea and / or seizures within 45 minutes, and make the subject insensitive within two hours. At levels of about 1000 ppm, an organism can expire after exposure for more than about 1 to 2 minutes. Due to the well-known and well-documented toxic effects of carbon monoxide for an organism, it is thus surprising and unexpected that carbon monoxide can be used to induce stasis of living biological samples and / or to help preserve them. Thus, it is contemplated that carbon monoxide can be used for the induction of stasis in isolated biological material, such as biological material free of blood (due to the effects that carbon monoxide has with respect to hemoglobin, which is a separate path different from the one that intervenes in the induction of stasis). In addition to exposure to carbon monoxide to induce stasis or to limit or prevent any damage caused by a stasis-inducing agent, the invention contemplates that carbon monoxide may be used in combination with agents or methods that assist in the preservation and preservation process. / or transplant / grafting of biological materials.

B. Chalcogenide compounds Compounds that contain a chalcogen element, those of group 6 of the periodic table, but which exclude oxides, are commonly referred to as "chalcogenides" or "chalcogenide compounds (used reciprocain the present.) These elements are sulfur ( S), selenium (Se), tellurium (Te) and polonium (Po) .Half-co-commons contain one or more of S, Se and Te, in addition to other elements.Charlcogenide compounds can be used as reducing agents. not limited by the following theory, thinks that the ability of chalcogenides to induce stasis in cells, and to allow the modulation of the temperature of the central part of the body in animals, is based on the union of these molecules to cytochrome oxidases. If so, chalcogenides inhibit or reduce the activity of oxidative phosphorylation. It is thought that the ability of chalcogenides to block autonomous thermoregulation, that is, to allow temperatures in the central part of the body of "warm-blooded" animals to be manipulated through the control of ambient temperatures, is based on the same mechanism discussed above - binding to cytochrome oxidase, and blocking or reducing the activity of oxidative phosphorylation. Chalcogenides can be supplied in liquid forms, as well as gaseous forms. Chalcogenides can be toxic, and at certain lethal levels, for mammals. In accordance with the present invention, it is anticipated that chalcogenide levels should not exceed lethal levels in the proper environment. Fatal levels of chalcogenides can be found, for example, in material safety data sheets for each chalcogenide, or from information sheets available from the Occupational Safety and Health Administration (OSHA) of the United States government. Although carbon monoxide and chalcogenide compounds can induce stasis by acting as an oxygen antagonist, they have different toxic effects that are separated from their ability to induce stasis. In addition, the concentrations needed to mediate a stasis effect are different, due to the different affinities of the cytochrome oxidase. While the affinity of cytochrome oxidase for oxygen is about 1: 1 compared to carbon monoxide, the affinity for H2S appears to be of the order of about 300: 1 compared to oxygen. This firmly fixes what toxic effects are observed with a concentration that induces stasis. In this way, it is contemplated that chalcogenide compounds are particularly suitable for the induction of stasis of biological matter in whole organisms and whole organisms. It can also prove useful in providing additional stimuli to a biological material before the chalcogenide is removed. In particular, it is envisioned that an animal can be subjected to an increased room temperature before the chalcogenide source is removed. 1. H2S Hydrogen sulfide (H2S) is a potentially toxic gas that is often associated with petrochemical and natural gas, sewage, paper pulp, leather tanning and food processing. The primary effect, at the cellular level, appears to be inhibition of cytochrome oxidase and other oxidative enzymes, resulting in cellular hypoxia. Exposure to extreme levels (500 ppm) results in sudden collapse and unconsciousness, a so-called "knock-down" effect, followed by recovery. Post-exposure effects may persist for years, and include loss of coordination, memory loss, motor dysfunction, personality changes, hallucination and insomnia. However, most contact with H2S occurs well below these acute toxicity levels. However, there is general interest in long-term contact at subacute levels. There are some reports that indicate persistent impairments in balance and memory, as well as altered motor sensory functions that can occur in humans after chronic exposure to low levels of H2S. See Kilburn and Warshaw (1995); Kilburn (1999). Others have reported that perinatal exposure of rats at low levels (20 or 50 ppm) of H2S for 7 hours per day of gestation through day 21 post-natal, resulted in longer dendritic branches with reduced arborization of Purkinje cells cerebelares. Other neurological defects associated with relatively low levels of H2S, include altered brain neurotransmitter concentrations, and altered neurological responses, such as increased theta hippocampal EEG activity. Functional toxicity was studied in rats exposed to moderate levels of H2S. The results showed that the H2S inhibits discriminated abstinence responses immediately after the end of the exposure (Higuchi and Fukamachi, 1997), and also interferes with the ability of rats to learn a primed task in the labyrinth of radial arms (Partió et al ., 2001). In another perinatal study using 80 ppm H2S, no neuropathological effects or altered motor activity, passive abstinence or startle acoustic response were observed in exposed rat pups. See Dorman et al., (2000). Finally, Struve ei al. (2001) exposed rats to H2S by gas at various levels for 3 hours per day in five consecutive days. Significant reductions in motor activity, water maze performance and body temperature were observed after exposure to 80 ppm H2S or more. Taken together, these reports indicate that H2S may have a variety of effects on the biochemistry of mammalian tissues, but there is no clear response pattern in terms of behavior. Typical levels of hydrogen sulfide contemplated for use in accordance with the present invention include values of about 1 to about 150 ppm, about 10 to about 140 ppm, about 20 to about 130 ppm and about 40 to about 120 ppm, or the oral, intravenous or transdermal equivalent dosage thereof. Other relevant scales include about 10 to about 80 ppm, about 20 to about 80 ppm, about 10 to about 70 ppm, about 20 to about 70 ppm, about 20 to about 60 ppm and about 30 to about 60 ppm, or oral dosage , intravenous or transdermal equivalent thereof. It is also contemplated that, for a given animal in a given period, the chalcogenide atmosphere should be reduced to avoid a potentially lethal accumulation of chalcogenide in the subject. For example, an initial environmental concentration of 80 ppm can be reduced after 30 minutes to 60 ppm, followed by further reductions at 1 hour (40 ppm) and 2 hours (20 ppm). 2. H? Se, H? Te and H? Po Hydrogen selenide (H2Se) is a key metabolite, formed from inorganic sodium selenite (oxidation state +4) via selenodiglutation (GSSeSG), through reduction by thiols and reductases dependent on NADPH, and released from selenocysteine by lyase action (Ganther, 1999). Hydrogen selenide provides selenium for the synthesis of selenoproteins after activation to selenophosphate. Hydrogen telluride (H2Te) exists as an unstable gas. 3. Other chalcogenides In certain embodiments, the reducing agent structure compound is dimethyl sulfoxide (DMSO), dimethyl sulfide (DMS), methyl mercaptan (CH3SH), mercaptoethanol, thiocyanate, hydrogen cyanide, methanethiol (MeSH) or CS2. In particular embodiments, the oxygen antagonist is CS2, MeSH or DMS. Particularly contemplated are compounds of the order of the size of these molecules (ie, within about 50% of their molecular weights). Additional compounds that are contemplated as being useful for the induction of stasis include, but are not limited to, the following structures, many of which are readily available-easily and are known to those skilled in the art (identified by CAS number): 104376 -79-6 (sodium salt of ceftriaxone); 105879-42-3; 1094-08-2 (etopropatine hydrochloride); 1098-60-8 (triflupromazine hydrochloride); 111974-72-2; 113-59-7; 113-98-4 (penicillin G potassium); 115-55-9; 1179-69-7; 118292-40-3; 119478-56-7; 120138-50-3; 121123-17-9; 121249-14-7; 1229-35-2; 1240-15-9; 1257-78-9 (prochlorperazine edisilate salt); 128345-62-0; 130-61-0 (thioridazine hydrochloride); 132-98-9 (penicillin V potassium); 13412-64-1 (dicloxacillin sodium hydrate); 134678-17-4; 144604-00-2; 146-54-3; 146-54-5 (fluphenazine dihydrochloride); 151767-02-1; 159989-65-8; 16960-16-0 (fragment 1-24 of adrenocorticotropic hormone); 1982-37-2; 21462-39-5 (clindamycin hydrochloride); 22189-31-7; 22202-75-1; 23288-49-5 (Probucol); 23325-78-2; 24356-60-3 (cephapirin); 24729-96-2 (clindamycin); 25507-04-4; 26605-69-6; 27164-46-1 (sodium cefazolin); 2746-81-8; 29560-58-8; 2975-34-0; 32672-69-8 (mesoridazine benzenesulfonate); 32887-01-7; 33286-22-5 ((+) - cis -diltiazem hydrochloride); 33564-30-6 (cefoxitin sodium); 346-18-9 3485-14-1; 3511-16-8; 37091-65-9 (sodium aziocillin); 37661-08-8; 3819-00-9 38821-53-3 (cephradine); 41372-02-5; 42540-40-9 (cefamandole naphtha) 4330-99-8 (heme salt - (+) - trimeprazine tartrate); 440-17-5 (trifluoperazine dihydrochloride); 4697-14-7 (disodium ticarcillin); 4800-94-6 (disodium carbenicillin); 50-52-2; 50-53-3; 5002-47-1; 51481-61-9 (cimetidine); 52239-63-1 (6-propyl-2-thiouracil); 53-60-1 (promazine hydrochloride); 5321-32-4; 54965-21-8 (albendazole); 5591-45-7 (thiothixene); 56238-63-2 (sodium cefuroxime); 56796-39-5 (sodium cefmetazole); 5714-00-1; 58-33-3 (promethazine hydrochloride); 58-38-8; 58-39-9 (perphenazine); 58-71-9 (sodium cephalothin); 59703-84-3 (sodium piperacillin); 60-99-1 (salt of methotrimeprazine maleate); 60925-61-3; 61270-78-8; 6130-64-9 (hydrated procaine penicillin G salt); 61318-91-0 (sulconazole nitrate salt); 61336-70-7 (amoxicillin trihydrate); 62893-20-3 (cefoperazone sodium); 64485-93-4 (cefotaxime sodium); 64544-07-6; 64872-77-1; 64953-12-4 (sodium moxalactam); 66104-23-2 (salt of pergolide mesylate); 66309-69-1; 66357-59-3 (ranitidine hydrochloride); 66592-87-8 (cefodroxil); 68401-82-1; 69-09-0 (chlorpromazine hydrochloride); 69-52-3 (ampicillin sodium); 69-53-4 (ampicillin); 69-57-8 (penicillin G sodium); 70059-30-2; 70356-03-5; 7081-40-5; 7081-44-9 (aqueous sodium cloxacillin); 7177-50-6 (aqueous sodium nafcillin); 7179-49-9; 7240-38-2 (aqueous sodium oxacillin); 7246-14-2; 74356-00-6; 74431-23-5; 74849-93-7; 75738-58-8; 76824-35-6 (famotidine); 76963-41-2; 79350-37-1; 81129-83-1; 84-02-6 (prochlorperazine dimaleate salt); 87-08-1 (phenoxymethylpenicillanic acid); 87239-81-4; 91-33-8 (benzthiazide); 91832-40-5; 94841-17-5; 99294-94-7; 154-42-7 (6-thioguanine); 36735-22-5; 536-33-4 (ethionamide); 52-67-5 (D-penicillamine); 304-55-2 (meso-2,3-dimercaptosuccinic acid); 59-52-9 (2,3-dimercapto (+) propanol); 6112-76-1 (6-mercaptopurine); 616-91-1 (N-acetyl-L-cysteine); 62571-86-2 (captopril); 52-01-7 (spironolactone); and 80474-14-2 (fluticasone propionate).

D. Other antagonists 1. Hypoxia and anoxia Hypoxia is a common natural tension, and there are several well-preserved responses that facilitate cellular adaptation to hypoxic environments. To compensate for the decrease in capacity for the production of aerobic energy in hypoxia, the cell must increase the production of anaerobic energy or decrease the demand for energy (Hochachka et al., 1996). Examples of these responses are common in metazoans, and the particular response used depends, in general, on the amount of oxygen available to the cell. In moderate hypoxia, oxidative phosphorylation is still partially active, so that some production of aerobic energy is possible. The cellular response to this situation, which is mediated in part by the hypoxia-inducible transcription factor, HIF-1, is to supplement the production of reduced aerobic energy by up-regulating genes involved in the production of anaerobic energy, such as glycolytic enzymes and transporters. of glu (Semenza, 2001; Guillemin et al., 1997). This response also promotes the upregulation of antioxidants such as catalase and superoxide dismutase, which protect against damage induced by free radicals. As a result, the cell is able to maintain almost normoxic activity levels in moderate hypoxia. In an extreme form of hypoxia, referred to as "anoxia" -defined here as < 0.001 kPa of 02 -, the oxidative phosphorylation ceases, and in this way the capacity to generate energy is drastically reduced. To survive in this environment, the cell must decrease the demand for energy by reducing cell activity (Hochachka et al., 2001). For example, in turtle hepatocytes deprived of oxygen, a cell-directed effort to limit activities such as protein synthesis, activity of ion channels and anabolic pathways, results in a 94% reduction in the demand for ATP (Hochachka ei al., 1996). In zebrafish embryos (Danio rerio), exposure to anoxia leads to complete arrest of heartbeat, movement, cell cycle progression and progression of development (Padilla et al., 2001). Likewise, C. elegans responds to anoxia by entering suspended animation, in which all observable movement ceases, including cell division and progression of development (Padilla et al., 2002, Van Voorhies et al., 2000). C. elegans can remain in suspended animation for 24 hours or more and, after returning to normoxia, will recover with high viability. This response allows C. elegans to survive hypoxic stress, decreasing the speed of energy-costly processes, and preventing the occurrence of damage, irreversible events such as aneuploidy (Padilla et al., 2002; Nystul et al., 2003). A newly discovered response is the generation of carbon monoxide induced by hypoxia by hem oxygenase-1 (Dulak et al., 2003). Carbon monoxide produced endogenously can activate signaling cascades that mitigate hypoxic damage through anti-apoptotic (Brouard et al., 2003) and anti-inflammatory (Otterbein et al., 2000) activity, and similar cryoprotective effects can be achieved in transplant models by perfusion with exogenous carbon monoxide (Otterbein et al., 2003; Amersi et al., 2002). At higher concentrations, carbon monoxide competes with oxygen for binding to iron-containing proteins, such as mitochondrial cytochromes and hemoglobin (Gorman et al., 2003), although the cytoprotective effect that this activity may have on hypoxia, does not It has been investigated. Despite the existence of these sophisticated defense mechanisms against hypoxic damage, hypoxia is still often a damaging strain. For example, mammals have hem oxygenase-1 and HIF-1, and some evidence suggests that suspended animation in mammals is also possible (Bellamy et al., 1996; Alam et al., 2002). However, hypoxic damage due to trauma such as heart attack, stroke or loss of blood is a major cause of death. Understanding the limitations of the two fundamental strategies for surviving hypoxic stress, remaining animated or in suspension animation, is impeded by the fact that it has been based on studies in a variety of systems under a variety of conditions. "Hypoxia" occurs when normal physiological levels of oxygen are not delivered to a cell or tissue. "Normoxia" refers to normal physiological levels of oxygen for the cell type, cell condition or tissue in particular. "Anoxia" is the absence of oxygen. "Hypoxic conditions" are those that lead to cellular hypoxia; These conditions depend on the type of cell, and the specific architecture or position of a cell within a tissue or organ, as well as the metabolic state of the cell. For purposes of the present invention, hypoxic conditions include conditions in which the oxygen concentration is at, or is less than, normal atmospheric conditions, i.e., less than 20.8, 20, 19, 18, 17, 16 , 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0%; alternatively, these numbers could represent the atmosphere percent at pressure 1 atmosphere (101.3 kPa). A zero percent oxygen concentration defines anoxic conditions. In this way, hypoxic conditions include anoxic conditions, although in some modalities, hypoxic conditions of no less than 0.5% are implemented. As used herein, "normoxic conditions" constitute oxygen concentrations of about 20.8% or greater. Standard methods to achieve hypoxia or anoxia are well established, and include the use of environmental chambers that depend on chemical catalysts that remove oxygen from the chamber. Said chambers are commercially available, for example, from BD Diagnostic Systems (Sparks, MD), as disposable casings of hydrogen + GASPAK carbon dioxide or BIO-BAG environmental chambers. Alternatively, oxygen may be depleted by exchanging air in a chamber with a gas other than oxygen, such as nitrogen. The oxygen concentration / concentration can be determined, for example, using an FYRITE oxygen analyzer (Bacharach, Pittsburgh PA). It is contemplated that the methods of the invention may use a combination of exposure to oxygen antagonists and alteration of oxygen concentrations, as compared to ambient air. In addition, the oxygen concentration of the environment containing the biological matter can be approximately, at least approximately, or at most approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 , 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37 , 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62 , 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87 , 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or 100%, or any scale derivable in these numbers. In addition, it is contemplated that a change in concentration may be any of the percentages or scales above, in terms of a decrease or increase compared to ambient air or a controlled environment. lll. Therapeutic or preventive applications A. Trauma In certain modalities, the present invention may find use in the treatment of patients who are suffering from trauma, or who are susceptible to it. Trauma can be caused by external trauma, such as burns, wounds, amputations and injuries from a firearm, or surgical trauma, internal trauma, such as stroke or heart attack resulting in acute reduction in circulation, or reductions in circulation due to at non-invasive tension, such as exposure to cold or radiation. On a cellular level, trauma often results in exposure of cells, tissues and / or organs to hypoxia, resulting in the induction of programmed cell death, or "apoptosis". Systemically, trauma leads to the induction of a series of biochemical processes, such as coagulation, inflammation or hypotension, and can lead to shock, which if persisted, can lead to organ dysfunction, irreversible cell damage, and death. Biological processes are designed to defend the body against traumatic attacks; however, they can lead to a sequence of events that proves to be dangerous and, in some cases, fatal. Therefore, the present invention contemplates the laying of tissues, organs, extremities and even entire organisms in stasis, as a way of protecting them from the damaging effects of the trauma. In a specific scenario, where medical care is not readily available, the induction of stasis in vivo or ex vivo, alternatively together with reduction in the temperature of the tissue, organ or organism, can "gain time" for the subject , taking medical attention to the subject, or transporting the subject to medical attention. The present invention also contemplates methods for the induction of tissue regeneration and wound healing, by preventing / delaying biological processes that can result in wound healing and regeneration of delayed tissues. In this context, in scenarios in which there is a substantial wound to the limb or organism, the induction of stasis in vivo or ex vivo, alternatively in conjunction with reduction in the temperature of the tissue, organ or organism, can help in the process of tissue regeneration and wound healing, controlling the biological processes that inhibit healing and regeneration. In addition to the wound healing and hemorrhagic shock that is discussed below, methods of the invention that prevent or treat trauma such as cardiac arrest or stroke can be implemented. The invention is of particular importance with respect to the risk of trauma by emergency surgical procedures, such as thoracotomy, laparotomy and splenic transection. 1. Wound healing In many cases, wounds and tissue damage are intractable, or take excessive periods to heal. Examples are chronic open wounds (diabetic foot ulcers and stage 3 and 4 pressure ulcers), acute and traumatic wounds, flaps and grafts, and subacute wounds (ie, open incisions). This can also be applied to other tissue damage, for example, burns and lung damage from smoking or from inhalation of hot air. Previous experiments show that hibernation to be protective against injury (for example, nails in the brain), can therefore have healing effects. Therefore, this technology can be useful in the control of wound healing processes, putting the tissue in a metabolically controlled environment. More particularly, the length of time that the cells or tissues are maintained in stasis can vary, depending on the injury. In some embodiments of the invention, the biological material is exposed to an oxygen antagonist for about at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, or more. 2. Hematological chogue (hemorrhagic chogue) This is a condition of profound hemodynamic and metabolic disorder characterized by the impossibility of the circulatory system to maintain adequate perfusion of vital organs. It can result from inadequate blood volume (hypovolemic shock), inadequate cardiac function (cardiogenic shock) or inadequate vasomotor tone (neurogenic shock, septic shock). This often results in rapid patient mortality. Whole body hibernation was induced in mice, and there was an immediate decrease in the overall metabolic state (measured by the production of C02). This was reversible, and the mice appeared to function normally, even after repeated exposures. Accordingly, the invention relates to the induction of a whole body hibernate state using H2S (or another oxygen antagonist), to preserve the life and vital organs of the patient. This will allow transportation to a controlled environment (eg, surgery), where the initial cause of the shock can be recorded, and then the patient can be brought back to normal function in a controlled manner. For this indication, the first hour after the injury, referred to as the "golden hour", is crucial for a successful outcome. The stabilization of the patient in this period is the main purpose, and the transport to a critical care unit (for example, emergency room, surgery, etc.), where the injury can be appropriately recorded. In this way, it would be ideal to keep the patient in stasis to allow this and record immediate problems, such as source of shock, supply of blood loss and reestablishment of homeostasis. Although this will vary significantly, in most cases, the amount of time that stasis will remain is between approximately 6 and approximately 72 hours after the injury. In some embodiments of the invention, the biological material is exposed to an oxygen antagonist for about at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 , 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days or more, and any scale or combination in these numbers.

B. Hypothermia In another embodiment, the present inventor proposes the use of the present invention to treat persons with extreme hypothermia. -The method provides that patients with extreme hypothermia are administered an oxygen antagonist or are exposed to it, and are then brought gradually to normal temperature while withdrawing, in a controlled manner, the oxygen antagonist. In this way, the oxygen antagonist dampens the biological systems within the subject, so that they can be started gradually without shock (or damage) to the subject. In one embodiment, a subject suffering from hypothermia may be given an oral or intravenous dose of an oxygen antagonist. Intravenous provision may be preferred due to the lack of potential sensitivity of the subject, and the ability to provide a controlled dosage over a period. Alternatively, if available, the oxygen antagonist can be provided in a gaseous state, for example, by using a mask for inhalation or even a sealed chamber that can accommodate the entire subject. Ideally, the patient will be stabilized in terms of heart rate, respiration and temperature before any change is made. Once stable, the ambient temperature will increase, again gradually. This can be achieved simply by removing the subject from hypothermic conditions. A more regulated increase in temperature can be achieved by adding successive layers of clothing or blankets, by using a thermal wrap with gradual increase in heat, or if possible, by placing the subject in a chamber whose temperature can be gradually increased. It is preferred that the subject's vital signs are monitored during the course of the temperature increase. Likewise, in conjunction with the increase in temperature, the oxygen antagonist is removed from the environment of the subject. Treatment with the oxygen antagonist and heat is interrupted at the appropriate endpoint, judged by the medical staff monitoring the situation, but in any case at the time when the subject's temperature and other vital signs return to a normal scale . Continuous monitoring is recommended after cessation of treatment for a period of at least 24 hours.

C. Hyperthermia Under certain conditions, which may result from genetic, infectious, pharmacological or environmental causes, patients may lose homeostatic temperature regulation resulting in severe uncontrollable fever (hyperthermia). This can result in long-term mortality or morbidity, especially brain damage, if not adequately controlled. Mice that inhaled H2S at 80 ppm, were immediately hibernating. This included an inability to regulate their body temperature when ambient temperatures were decreased below room temperature. Therefore, this technology could be used to control the temperature of the whole body in certain states of hyperthermia.

This would probably include the administration of H2S (or another oxygen antagonist) through inhalation or perfused into the bloodstream to induce a state of hibernation. It would be useful to have the patient stasis for between about 6 and about 24 hours, during which time the source of the fever can be reported. In some embodiments of the invention, a patient is exposed to an oxygen antagonist for about at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30 , 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days or more, and any scale or combination in these numbers. This can be combined with some regulation of the whole body temperature (ice bath / blanket / cooling system).

D. Cardioplegia In certain embodiments, the present invention may find use as solutions for cardioplegia for cardiac bypass surgery (CABG). Cardioplegic solutions are perfused through the vessels and chambers of the heart, and cause their intrinsic beat to cease, while maintaining organ viability. Cardioplegia (heart paralysis) is desirable during open-heart surgery, and during the procurement, transportation and storage of donor hearts for use in cardiac transplant procedures.

Several different cardioplegic solutions are available, and different techniques for the use of cardioplegic solutions are known in the art. For example, cardioplegic solutions often have varying amounts of potassium, magnesium and several other minor components. Sometimes drugs are added to the cardioplegic solution to aid in muscle relaxation and protection from ischemia. Changing the temperature at which the cardioplegic solution is used can also have beneficial effects. Despite the protective effects provided by common methods for the induction of cardioplegia, there is still some degree of ischemic-reperfusion injury to the myocardium. Ischemic-reperfusion injury during cardiac bypass surgery results in poor outcomes (both morbidity and mortality), especially due to the already weakened state of the heart. Myocardial ischemia results in anaerobic myocardial metabolism. The final products of anaerobic metabolism rapidly lead to acidosis, mitochondrial dysfunction and myocyte necrosis. High energy phosphate depletion occurs almost immediately, with a 50% loss of ATP stores within 10 minutes. Reduced contractility occurs within 1 to 2 minutes, with development of ischemic contracture and irreversible injury after 30 to 40 minutes of normothermic ischemia (37 ° C). Reperfusion injury is a well-known phenomenon that follows the restoration of the coronary circulation. Reperfusion injury is characterized by abnormal oxidative myocardial metabolism. In addition to structural changes created during ischemia, reperfusion can produce cytotoxic oxygen free radicals. These oxygen-free radicals play an important role in the pathogenesis of reperfusion injury, oxidizing the phospholipids of the sarcolemma, and in this way disorganizing the integrity of the membrane. Oxidized free fatty acids are released into the coronary venous blood, and are a marker of peroxidation of the phospholipids of the myocardial membrane. Protamine induces complement activation, which activates neutrophils. Activated neutrophils and other leukocytes are an additional source of oxygen free radicals and other cytotoxic substances. The present invention provides methods and compositions for the induction of cardioplegia that will provide greater protection to the heart during bypass surgery. In certain embodiments, the present invention provides a cardioplegic solution comprising H2S dissolved in solution (or other oxygen antagonist). In some embodiments, the invention further comprises at least a first device, such as a catheter or cannula, for introducing an adequate dose of the cardioplegic solution to the heart. In certain aspects, the invention further comprises at least a second device, such as a catheter or cannula, for removal of the cardioplegic solution of the heart. Typically, bypass surgery takes 3 to 6 hours; however, complications and CABG of multiple vessels can extend the duration to 12 hours or more. It is contemplated that the heart be maintained in stasis during surgery. Thus, in some embodiments of the invention, the heart is exposed to an oxygen antagonist for about at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20 , 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 hours or more, and any scale or combination in these numbers.

E. Harm reduction by cancer therapy Cancer is a leading cause of mortality in industrialized countraround the world. The most conventional procedure for the treatment of cancer is administering a cytotoxic agent to the cancer patient (or ex vivo treatment of a tissue), so that the agent has a more lethal effect on the cancer cells than on the normal cells. The greater the dose or the more lethal the agent is, the more effective it will be; however, for the same reason, these agents are those that are more toxic (and sometimes lethal) to normal cells. Therefore, chemotherapy and radiation therapy are often characterized by severe side effects, some of which are life-threatening, for example, ulcers in the mouth, difficulty swallowing, dry mouth, nausea, diarrhea, vomiting, fatigue, hemorrhage, hair loss and infection, skin irritation and loss of energy (Curran, 1998; Brizel, 1998). Recent studsuggest that the transient and reversible decrease in the temperature of the central part of the body, or "hypothermia", can lead to improvements in the fight against cancer. It was recently found that hypothermia of 28 ° C reduces the toxicity induced by radiation, doxorubicin and cisplatin in mice. The anti-cancer activity of these drugs / treatments was not compromised when administered to cold-treated animals; rather, it improved, particularly for cisplatin (Lundgren-Erikkson et al., 2001). Based on this and other published works, the inventor proposes that a further reduction in the temperature of the central part of the body will provide benefit to patients with cancer. Thus, the present invention contemplates the use of oxygen antagonists to induce stasis in normal tissues of a cancer patient, thereby reducing the potential impact of chemotherapy or radiotherapy on those tissues. It also allows the use of higher doses of chemotherapy and radiotherapy, thus increasing the anticancer effects of these treatments. The treatment of virtually any hyperproliferative disorder, including benign and malignant neoplasms, non-neoplastic hyperproliferative conditions, preneoplastic conditions and precancerous lesions is contemplated. Such disorders include restenosis, cancer, cancer resistant to multiple drugs, primary psoriasis and metastatic tumors, angiogenesis, rheumatoid arthritis, inflammatory bowel disease, psoriasis, eczema and secondary cataracts, as well as oral hairy leukoplakia, bronchial dysplasia, carcinomas in situ, and intraepithelial hyperplasm. In particular, the present invention is directed to the treatment of human cancers that include cancers of prostate, lung, brain, skin, liver, breast, lymphoid system, stomach, testes, ovar pancreas, bone, bone marrow, gastrointestinal system, head and neck, cervix, esophagus, eye, gall bladder, kidney, suprarenal glands, heart, colon and blood. Cancers involving epithelial and endothelial cells are also contemplated for treatment. In general, chemotherapy and radiotherapy are designed to reduce the size of the tumor, reduce the growth of tumor cells, induce apoptosis in tumor cells, reduce tumor vasculature, reduce or prevent metastasis, reduce the rate of tumor growth, accelerate the death of tumor cells, and destroy tumor cells. The purposes of the present invention are not different. Thus, it is contemplated that oxygen antagonist compositions of the present invention will be combined with secondary anticancer agents (side agents) effective in the treatment of hyperproliferative disease. An "anti-cancer" agent is capable of negatively affecting the cancer in a subject, for example by destroying cancer cells, inducing apoptosis in cancer cells, reducing the rapid growth of cancer cells, reducing the incidence or the number of metastases, reducing the size of the tumor, inhibiting tumor growth, reducing the blood supply to a tumor or cancer cells, promoting a immune response against cancer cells or a tumor, preventing or inhibiting the progression of cancer, or increasing the lifespan of a subject with cancer. Secondary anticancer agents include biological agents (biotherapy), chemotherapeutic agents and radiotherapeutic agents. More generally, these other compositions are provided in a combined amount effective to destroy or inhibit the proliferation of cancer or tumor cells, while at the same time reducing or minimizing the impact of the secondary agents on normal cells. This method may include contacting or exposing the cells with an oxygen antagonist and the secondary agents at the same time. This may be achieved by contacting the cell with an individual drug formulation or formulation that includes both agents, or by contacting or exposing the cell with two different compositions or formulations, at the same time, wherein a composition includes an oxygen antagonist, and the other includes the secondary agents. Alternatively, oxygen antagonist therapy may precede or continue treatment with the secondary agent at intervals ranging from minutes to weeks. In modalities where the other agent and expression construction are applied separately to the cell, it would be generally ensured that a significant period does not expire between the time of each supply, so that the agent and expression construction are still capable of exerting an advantageously combined effect on the cell. In such cases, it is contemplated that the cell can be contacted with both modalities within about 12 to 24 hours of each other and, more preferably, within about 6 to 12 hours of each other. In some situations, it may be desirable to significantly extend the treatment period, however, where several days (2, 3, 4, 5, 6 or 7) to several weeks (1, 2, 3, 4, 5, 6, 7 or 8) pass between the respective administrations. In certain embodiments, it is anticipated that the biological material will be maintained in stasis for between about 2 and about 4 hours, while the cancer treatment is being administered. In some embodiments of the invention, the biological material is exposed to an oxygen antagonist for about at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6 hours or more, and any scale or combination in these numbers. Various combinations can be used; the oxygen antagonist is "A", and the secondary anticancer agent, such as radiotherapy or chemotherapy, is "B": A / B / A B / A B B / B / A PJ A / B A / B B B / A A A B / B / B B / A / B / B B / B / B /? B / B / A / BA / AJB / BA / B / A / BAB / B / AB / B / A / AB / A / B / AB / A / A / BA / AA / BB / A / A / AA / B / AAAAB / A l The administration of the oxygen antagonist compounds of the present invention to a patient will follow general protocols for the administration of chemotherapeutics, taking into account the toxicity of the compound, if any. It is expected that the treatment cycles are repeated as necessary. It is also contemplated that various standard therapies, as well as surgical intervention, may be applied in combination with the anticancer therapy described above. 1. Chemotherapy Cancer therapies also include a variety of combination therapies with treatments based on chemicals and radiation. Combination chemotherapies include, for example, cisplatin (CDDP), carboplatin, procarbazine, mecloretamine, cyclophosphamide, camptothecin, ifosfamide, melphalan, chlorambucil, busulfan, nitrosourea, dactinomycin, daunorubicin, doxorubicin, bleomycin, plicamycin, mitomycin, etoposide (VP16). , tamoxifen, raloxifene, estrogen receptor binding agents, taxol, gemcitabine, navelbine, farnesyl-protein transferase inhibitors, transplatino, 5-fluorouracil, vincristine, vinblastine and methotrexate, temazolomide (an aqueous form of DTIC), or any analogue or derivative or variant of the above. The combination of chemotherapy with biological therapy is known as biochemotherapy. 2. Radiation therapy Other factors that cause DNA damage and that have been used extensively include those that are commonly known as gamma rays, X-rays, and / or the targeted delivery of radioisotopes to tumor cells. Other forms of factors that damage DNA are also contemplated, such as microwave and UV irradiation. All these factors are more likely to effect a wide range of DNA damage, in DNA precursors, in DNA replication and repair, and in the assembly and maintenance of chromosomes. Dosage scales for X-rays vary from daily doses of 50 to 200 Roentgens for prolonged periods (3 to 4 weeks), up to individual doses of 2000 to 6000 Roentgens. Dosage scales for radioisotopes vary widely, and depend on the half-life of the isotope, the concentration and type of radiation emitted, and the uptake by the neoplastic cells. The terms "in contact" and "exposed", when applied to a cell, are used herein to describe the process by which a composition of the invention (e.g., a hypoxic antitumor compound) or a chemotherapeutic or radiotherapeutic agent it is supplied to a target cell, or they are placed in direct juxtaposition with the target cell. In combination therapy, to achieve cell or stasis destruction, both agents can be delivered to a cell in an effective combined amount to destroy the cell or prevent it from dividing. 3. Immunotherapy Immunotherapeutics depends, in general, on the use of immune effector cells and molecules that they target and destroy cancer cells. The immune effector may be, for example, an antibody specific for a marker on the surface of a tumor cell. The antibody can only serve as a therapy effector, or it can recruit other cells that ultimately effect the destruction of the cells. The antibody can also be conjugated to a drug or toxin (chemotherapeutic, radionuclide, ricin A chain, cholera toxin, pertussis toxin, etc.), and serve only as an agent of choice of target. Alternatively, the effector may be a lymphocyte that possesses a surface molecule that interacts, directly or indirectly, with a target tumor cell. Several effector cells include cytotoxic T cells and NK cells. Immunotherapy could also be used as part of a combination therapy. The general procedure for combination therapy is discussed below. In an aspect of immunotherapy, the tumor cell must have a marker that is subject to the choice of target, that is, it is not present in most other cells. There are many tumor markers, and any of these may be suitable to function in the choice of target in the context of the present invention. Common tumor markers include carcinoembryogenic antigen, prostate specific antigen, urinary tumor-associated antigen, fetal antigen, tyrosinase (p97), gp68, TAG-72, HMFG, sialyl Lewis antigen, MucA, MucB, PLAP, receptor estrogen, laminin receptor, erb B and p155. An alternative aspect of immunotherapy is towards anticancer effects with immune stimulating effects. There are also immune stimulation molecules that include: cytokines such as IL-2, IL-4, IL-12, GM-CSF, gamma-IFN, chemokines such as MIP-1, MCP-1, IL-8 and growth factors such as the FLT3 ligand. It has been shown that the combination of immune stimulation molecules, either as proteins or using gene delivery in combination with a tumor suppressor such as mda-7, enhances the antitumor effects (Ju et al., 2000). As discussed initially, examples of immunotherapies currently under investigation or in use, are immune adjuvants (e.g., Mycobacterium bovis, Plasmodium falciparum, dinitrochlorobenzene and aromatic compounds) (see U.S. Patent No. 5,801, 005; patent of E.U.A. No. 5,739,169; Hui and Hashimoto, 1998; Christodoulides et al., 1998), therapy with cytokines (for example, interferons, β and β, IL-1, GM-CSF and FNT) (Bukowski et al., 1998; Davidson et al., 1998; Hellstrand et al. , 1998), gene therapy (eg, FNT, IL-1, IL-2, p53) (Qin et al., 1998; Austin-Ward and Villaseca, 1998; U.S. Patent No. 5,830,880 and U.S. Patent No. 8,846,945) and monoclonal antibodies (eg, anti-ganglioside GM2, anti-HER-2, anti-p185) (Pietras et al., 1998; Hanibuchi et al., 1998). Herceptin (trastuzumab) is a chimeric monoclonal antibody (mouse-human) that blocks the HER2-neu receptor. It has antitumor activity, and has been approved for use in the treatment of malignant tumors (Dillman, 1999). It has been shown that the combination therapy of -cancer with herceptin and chemotherapy is more effective than individual therapies.- Thus, it is contemplated that one or more anticancer therapies may be used with the antitumor therapies described herein.

F. Neurodegeneration The present invention can be used to treat neurodegenerative diseases. Neurodegenerative diseases are characterized by degeneration of neuronal tissue, and are often accompanied by memory loss, loss of motor function, and dementia. With dementia diseases, higher cognitive and integrative cognitive faculties become more and more deteriorated over time. It is estimated that approximately 15% of people aged 65 or older show mild to moderate dementia. Neurodegenerative diseases include Parkinson's disease; Primary neurodegenerative disease; Huntington's disease; apoplexy and other hypoxic or ischemic processes; neurotrauma; metabolically induced neurological damage; sequelae of cerebral convulsions; hemorrhagic shock; secondary neurodegenerative disease (metabolic or toxic); Alzheimer's disease, other memory disorders; or vascular dementia, dementia due to multiple infarctions, dementia of the Lewy body, or neurodegenerative dementia. The evidence shows that the health of an organism, and especially the nervous system, depends on the oscillation between oxidative and reductive states, which are intimately linked to circadian rhythms. That is, the oxidative stress placed on the body during consciousness oscillates towards a reductive environment during sleep. It is thought that this is a big part of why sleep is so important for health. Certain states of neurodegenerative disease, such as Huntlngton's disease and Alzheimer's disease, as well as normal aging processes, have been linked to a disagreement in this oscillation pattern. There is also some evidence that H2S levels in the brain are reduced under these conditions (Eto ef al., 2002). The present invention can be used to regulate and control the oscillation between the oxidative and reduced states, to prevent or reverse the effects of diseases and neurodegenerative processes. Furthermore, it has been shown in general that reduced metabolic activity correlates with health in aged animals and humans. Therefore, the present invention would also be useful in suppressing the general metabolic function that increases longevity and health in mature age. It is contemplated that this type of treatment would probably be administered at night, during sleep for a period of approximately 6 to 10 hours a day. This may require daily treatment for extended periods of months to years.

IV. Preservation applications The present invention can be used to preserve or store whole organisms for transport and / or storage purposes.

Such organisms could be used for research purposes, such as laboratory mice (creation of mouse banks), or for consumption, such as fish. In this situation, it is contemplated that stasis can be maintained indefinitely. In addition, stasis can be induced in plants or parts of plants, which include fruit, flowers, leaves, stems, seeds and cuttings. The plants can be agricultural, medicinal or decorative. The induction of stasis in plants can intensify the life of storage or the resistance of the whole plant or part of it to pathogens. Thus, in embodiments of the invention, an organism or part thereof can be exposed to an oxygen antagonist for about at least about, or at most about 30 seconds, 1, 2, 3, 4, 5, 10 , 15, 20, 25, 30, 35, 40, 45, 50, 55 minutes, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 hours, 1, 2, 3, 4, 5, 6, 7 days, 1, 2, 3, 4, 5 weeks, 1, 2, 3 , 4, 5, 6, 7, 8, 9, 10, 11, 12 months, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more years, and any combination or scale derivable in these numbers.

A. Other preservation agents A variety of preservation solutions have been described in which the organ is surrounded or perfused with the preservation solution while it is being transported. One of the most commonly used solutions is ViaSpan® (Belzer UW), which is used with cold storage. Other examples of such solutions or components of said solutions include the St. Thomas solution (Ledingham et al., J. Thorac, Cardiovasc. Surg. 93: 240-246, 1987), Broussais solution, UW solution (Ledingham et al. al., Circulation 82 (part 2) IV 351-8, 1990), Celsior solution (Menasche et al., Eur. J. Cardio, Thorax, Surg., 8: 207-213, 1994), solution from Stanford University and solution B20 (Bernard et al., J. Thorac. Cardiovasc.

Surg. 90: 235-242, 1985), as well as those described and / or claimed in the US patents. Nos. 6,524,785; 6,492,103; 6,365,338; 6,054,261; ,719,174; 5,693,462; 5,599,659; 5,552,267; 5,405,742; 5,370,989; 5,066,578; 4,938,961; and 4,798,824. In addition to solutions, other types of materials are also known for use in the transportation of organs and tissues. These include gelatinous material, as well as other semi-solid material, such as those described, for example, in U.S. Pat. No. 5,736,397. Some of the systems and solutions for the preservation of organs, specifically involve the perfusion of oxygen in the solution or system to expose the organ to oxygen, since it is thought that the maintenance of the organ or tissue in an oxygenated environment, improves its viability . See Kuroda et al. (Transplantation 46 (3): 457-460, 1988), and the patents of E.U.A. Nos. 6,490,880; 6,046,046; 5,476,763; 5,285,657; 3,995,444; 3,881, 990; and 3,777,507. It is thought that isolated hearts that are deprived of oxygen for more than four hours, lose vigor and are not useful in the recipient, due to ischemic / reperfusion injury. See the patent of E.U.A. No. 6,054,261. In addition, many of the containers and solutions, if not all, for the preservation and transplantation of organs, involve hypothermia (temperature below room temperature, often close to 0 ° C, but not below this temperature), which It has been called the "solid basis of all useful methods of organ and tissue preservation". See the patent of E.U.A. No. 6,492,103. To improve the landscape of a successful transplant, techniques have been developed to better preserve an organ for transplantation. Two general areas of development have occurred, one in the area of preservation solutions, and the other in the area of organ containers. In the field of organ transplantation, it is thought that certain conditions are related to the condition of the organ and the prognosis for a successful transplant: 1) minimization of edema and swelling of the cells; 2) prevention of intracellular acidosis; 3) Minimization of ischemic damage; and 4) provision of substrate for the regeneration of high energy phosphate compounds and ATP during reperfusion. Ischemic / reperfusion injury in organ transplantation is especially problematic because the harvested organ is removed from the body, isolated from a blood source, and thus deprived of oxygen and nutrients for an extended period (see patent from US No. 5,912,019). In fact, one of the most critical problems in transplantation today is the relatively high incidence of delayed graft function (DGF) due to acute tubular necrosis (ATN) after surgery. Current methods still experience problems in these areas, which highlights the importance of the present invention.

However, the present invention can be used in conjunction with other preservation compositions and methods. As discussed in the patents of E.U.A. Nos. 5,952,168, 5,217,860, 4,559,258 and 6,187,529 (incorporated herein by reference), biological materials can be preserved, for example, by maintaining for a long time transplantable or replaceable organs. Cells, tissues, organs or corpses can be given compounds that intensify or maintain the condition of the organs for transplantation. Said methods and compositions include those described in the U.S.A. Nos. 5,752,929 and 5,395,314. In addition, the methods of the present invention may include exposure of biological matter to preservation solutions, such as those discussed above, in addition to exposure to an oxygen antagonist. It is contemplated that any agent or solution that is used with a biological sample that is alive and that will be used as a living material, be pharmaceutically acceptable or pharmacologically acceptable. The phrase "pharmaceutically acceptable" or "pharmacologically acceptable" refers to molecular entities and compositions that do not produce an allergic or similar adverse reaction when administered to a human. The preparation of an aqueous composition containing a protein as an active ingredient is well known in the art. Typically, said compositions are prepared as suspensions or liquid solutions; solid forms suitable for solution in, or suspension in, liquid can also be prepared before use. Transplant organs can be monitored to assess their condition, particularly with respect to their use as a transplant. Said methods are described in the patent of E.U.A. No. 5,699,793. Many drugs can be administered to a patient after he receives an organ transplant to facilitate the recovery process. Such drugs include compounds and agents that reduce or inhibit an immune response against the donor organ. In addition, additional drugs are continually being investigated and offered for use in organ transplantation, such as those described in the U.S. Patents. No. 6,552,083 (inhibitory agent comprising N- (3,4-dimethoxycinnamoyl) anthranilyl acid) and 6,013,256 (antibodies that bind to the IL-2 receptor, such as a humanized anti-Tax antibody).

B. Preservation devices Systems or containers for the transport of organs and tissues have also been developed over the years. Any of these modalities can be combined with apparatuses of the invention, which allow the use with oxygen antagonists. Most include cooling systems for implementation, for example, those described in the US patents.

Nos. 4,292,817, 4,473,637 and 4,745,759, which employ active cooling with a cooling liquid that is pumped through the system. Several sophisticated devices have been designed including multiple chambers or double containers, as described in the U.S. Patents. Nos. 5,434,045 and 4,723,974. Some constitute a system in which an apparatus for perfusing the organ or tissue in a preservation solution is designed, as described in the U.S. Patents. Nos. 6,490,880; 6,100,082; 6,046,046; 5,326,706; 5,285,657; 5,157,930; 4,951, 482; 4,502,295; and 4,186,565.

V. Diagnostic applications Sulfites are produced by all cells in the body during the normal metabolism of sulfur-containing amino acids. Sulfite oxidase removes, and thus regulates, sulfite levels. The differential activities of these enzymes would lead to different levels of sulfites produced in tissue-specific form. In the example described above, for solid tumors under hypoxic conditions, sulfites can be produced at higher levels that provide a local protective state against tumor cells through the reduction of the metabolic state, as well as the inhibition of immune surveillance. Therefore, it would be beneficial to measure sulfite levels, and incorporate this as part of the diagnosis for various disease states such as solid tumors. In addition, since the use of sulphites is proposed for various applications, the use of some type of imaging procedure or other monitoring procedure would be useful thereafter. It is possible to measure sulphite levels in serum to obtain a total sulfite level through the use of common technology (for example, CLAR). It is useful to explore the possibility of sulfite image formation. Alternatively, a proteomic alternative may allow an understanding of how the regulation of enzymes involved in sulfite metabolism can be altered in certain disease states, which allows this alternative for diagnosis.

SAW. Applications in Selection In other embodiments, the present invention provides methods for the identification of oxygen antagonists and molecules that act in a similar manner with respect to the induction of stasis. In some cases, the oxygen antagonist being sought works as a chalcogenide compound, reducing the temperature of the central part of the body or preserving viability in hypoxic or anoxic environments that would otherwise destroy biological matter were it not for the presence of the oxygen antagonist. These tests may include random selection of large collections of candidate substances; alternatively, the tests can be used to focus on particular classes of selected compounds in search of attributes that are thought to make them more likely to act as oxygen antagonists. For example, a method generally comprises: (a) providing a candidate modulator; (b) mixing the candidate modulator with a biological material; (c) measuring one or more cellular responses characteristic of treatment with oxygen antagonists; and- (d) comparing the one or more responses with biological matter in the absence of the candidate modulator. Tests can be carried out on isolated cells, tissues and organs, or intact organisms. In fact, it will be understood that all selection methods of the present invention are useful in their own right, despite the fact that effective candidates may not be found. The invention provides methods for the selection of said candidates, not only methods to find them. However, it will also be understood that a modulator can be identified as an effective modulator according to one or more tests, indicating that the modulator appears to have some capacity to act as an oxygen antagonist, such as by inducing stasis in a biological material. The selection, in some modalities, involves the use of a test described in the examples to identify a modulator. An effective modulator can also be characterized or tested. In addition, the effective modulator can be used in an animal or animal model in vivo (as discussed below), or it can be used in other animals or animal models in vivo, which may include the same species of animals or in different animal species. Furthermore, it is contemplated that the modulator identified in accordance with the embodiments of the invention may also be manufactured after selection. Also, the biological material can be exposed to an effective modulator according to the methods of the invention, or it can be contacted therewith, in particular with respect to therapeutic or preservation modalities.

A. Modulators As used herein, the term "candidate substance" refers to any molecule that can induce stasis in biological matter, for example, by altering the temperature of the central part of the body. The candidate substance can be a protein or fragment thereof, a small molecule, or even a nucleic acid molecule. They can also be purchased, from several commercial sources, collections of small molecules that are thought to cover the basic criteria for useful drugs in an effort to achieve by "brute force" the identification of useful compounds. The selection of such collections, including combinatorially generated collections (eg, peptide collections), is a fast and efficient way of selecting a large number of related (and unrelated) compounds for activity. The combinatorial methods themselves also lead to the rapid production of potential drugs by the creation of second, third and fourth generation modeled compounds of active, but otherwise undesirable compounds. Candidate compounds may include fragments or parts of naturally occurring compounds, or may be found as active combinations of known compounds, which are otherwise inactive. It is proposed that compounds isolated from natural sources, such as animals, bacteria, fungi, plant sources, including leaves and bark, and marine sources, can be tested as candidates for the presence of potentially useful pharmaceutical agents. It will be understood that the pharmaceutical agents to be selected could be derived or synthesized also from chemical compositions or compounds synthesized by man. Thus, it is understood that the candidate substance identified by the present invention can be a peptide, polypeptide, polynucleotide, small molecule inhibitors or any other compound that can be designed through rational drug design, starting from known inhibitors or stimulators. Other suitable modulators include antisense molecules, siRNA molecules, ribozymes and antibodies (including single chain antibodies), each of which would be specific to the target molecule. Such compounds are described in more detail elsewhere in this document. For example, an antisense molecule that binds to a transcription or translation initiation site, or splicing junctions, would be an ideal candidate inhibitor.

In addition to the modulator compounds initially identified, the inventor also contemplates that other structurally similar compounds may be formulated that mimic the key portions of the structure of the modulators. Such compounds, which may include peptidomimetics of peptide modulators, can be used in the same way as the initial modulators.

B. In vitro tests In vitro tests include the use of several animal models. Due to their size, ease of handling and information on their physiology and genetic constitution, mice are a preferred modality. However, other animals are also suitable, and include rats, rabbits, hamsters, guinea pigs, gerbils, North American marmots, mice, cats, dogs, sheep, goats, pigs, cows, horses and monkeys (including chimpanzees, gibbons and baboons). Fish are also contemplated for use in in vivo tests, as well as nematodes. Tests for modulators can be carried out using an animal model derived from any of these species. In such tests, one or more candidate substances are administered to an animal, and the ability of the candidate substances to induce stasis, reduce the temperature of the central part of the body, or endow in the biological material the ability to survive hypoxic environmental conditions or anoxic, compared to an inert vehicle (negative control) and H2S (positive control), identify a modulator. Treatment of animals with test compounds will include administration of the compound, in a suitable form, to the animal. The administration of the candidate compound (gas or liquid) will be by any route that can be used for clinical or non-clinical purposes including, but not limited to, oral, nasal (inhalation or aerosol), buccal or even topical administration. Alternatively, administration can be by intratracheal instillation, bronchial instillation and intradermal, subcutaneous, intramuscular, intraperitoneal or intravenous injection. Pathways specifically contemplated are systemic intravenous injection, regional administration by means of blood or lymph supply, or directly to an affected site.

Vll. Modes of administration and pharmaceutical compositions An effective amount of a chalcogenide pharmaceutical composition is generally defined as the amount sufficient to improve, reduce, minimize or limit detectably the degree of the condition of interest. More rigorous definitions can be applied, including elimination, eradication or cure of the disease.

A. Administration Routes of administration of a chalcogenide will, of course, vary with the location and nature of the condition to be treated and include, for example, intradermal, transdermal, parenteral, intravenous, intramuscular, intranasal, subcutaneous, percutaneous, intratracheal, intraperitoneal, intratumoral, by perfusion, by inhalation, washing, direct injection and administration and oral formulation. In the case of transplantation, the present invention can be used pre-operatively and / or post-operatively to cause the host or graft materials to come to rest. In a specific embodiment, a surgical site can be injected or perfused with a formulation comprising a chalcogenide. Perfusion can be continued post-surgery, for example, leaving an implanted catheter at the site of surgery.

B. Injectable Formulations and Compositions Preferred methods for delivery of the oxygen antagonists of the present invention are inhalation, intravenous injection, perfusion of a particular area and oral administration. However, the pharmaceutical compositions described herein may alternatively be administered parenterally, intradermally, intramuscularly, transdermally or even intraperitoneally, as described in the US patent. No. 5,543,158; patent of E.U.A. No. 5,641, 515 and patent of E.U.A. No. 5,399,363 (each specifically incorporated herein by reference in its entirety). Solutions of the active compounds can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof, and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative that prevents the growth of microorganisms. Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or sterile dispersions and powders for the extemporaneous preparation of sterile injectable solutions or dispersions (see U.S. Patent No. 5,466,468, specifically incorporated herein by reference in its entirety). In all cases, the form must be sterile, and must be fluid to the extent that there is easy application by syringe. It must be stable under the conditions of manufacture and storage, and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (eg, glycerol, propylene glycol and liquid polyethylene glycol, and the like), suitable mixtures thereof, and / or vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. The prevention of the action of microorganisms can be achieved by the use of various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, - sorbic acid, thimerosal, and the like. In many cases, it will be preferred to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be achieved by the use, in the compositions, of agents that retard absorption, for example, aluminum monostearate and gelatin. For parenteral administration in an aqueous solution, for example, the solution should be adjusted to its pH properly if necessary, and the liquid diluent should first be made isotonic with enough saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous, intratumoral and intraperitoneal administration. In this regard, sterile aqueous media that can be used will be known to those skilled in the art in light of the present disclosure. For example, a dosage may be dissolved in 1 ml of isotonic NaCl solution, and it may be added to 1000 ml of hypodermoclysis fluid, or it may be injected at the proposed infusion site (see, for example, "Remington's Pharmaceutical Sciences", fifteenth edition). , pp. 1035-1038 and 1570-1580). Certain variation in the dosage will necessarily occur, depending on the condition of the subject being treated. The responsible of the administration will determine, in any case, the adequate dose for the individual subject. In addition, for human administration, the preparations must meet standards of sterility, pyrogenicity, general safety and purity, as required by FDA Office of Biologics standards. Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in the appropriate solvent, with several of the other ingredients listed above, as required, followed by filtered sterilization. In general, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle containing the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred preparation methods are vacuum drying and freeze drying techniques, which give a powder of the active ingredient plus any additional desired ingredients from a previously sterile filtered solution. of the same. As used herein, the term "carrier" includes any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic agents and absorption retarders, pH regulators, vehicle solutions, suspensions, colloids, and the like. The use of said media and agents for pharmaceutical active substances is well known in the art. Except where conventional means or agents are incompatible with the active ingredient, their use in the therapeutic compositions is contemplated. Complementary active ingredients can also be incorporated into the compositions.

C. Catheters In certain embodiments, a catheter is used to provide a protective agent to an organism. Of particular interest is the administration of said agent to the heart or vascular system. Often, a catheter is used for this purpose. Yaffe et al., 2004 discusses catheters in particular in the context of suspended animation, although the use of catheters was generally known before this publication.

D. Gas supply 1. Breathing System An example of a gas supply system 100 is illustrated in Figure 9. The delivery system 100 is suitable for the supply of respirable gases, including an active agent, to the respiratory system of a subject. The gas supply system 100 includes one or more gas sources 102. Each of the gas sources 102 is connected to a regulator 104 and a flow meter 106. The gas supply system 100 also includes a source of agent active 107, an optional vaporizer 108, an output controller 110, a scrubber 112, and an alarm / monitoring system 114. The delivery system 100 may include certain elements generally used in an anesthesia delivery machine. For example, anesthesia delivery machines generally include a high pressure circuit, a low pressure circuit, a breathing circuit and a purification circuit. As described in Figures 10 to 11, one or more of the gas sources 102, the vaporizer 108, the exit controller 110, the scrubber 112 and / or the alarm / monitoring system 114, may be provided as part of a device having a circuit of high pressure, low pressure, respiration and / or purification, and these elements may be similar to those generally used in an anesthesia delivery machine. Anesthesia delivery machines are described, for example, in the US patents. Nos. 4,034,753; 4,266,573; 4,442,856; and 5,568,910, the contents of which are hereby incorporated by reference in their entirety. The gas sources 102 can be provided by tanks of compressed gas; however, it should be understood that the gas sources 102 may be a gas or a source of liquid that is converted to a gas. For example, vaporizer 108 can be used to vaporize a source of liquid gas. Regulators 104 include valves that reduce the pressure of each of the gas sources 102. The decompressed gas then passes through one of the flow meters 106, which measures and controls the flow of gas from each of the sources of gas. 102 gas. The gas sources 102 can be vehicle gases that are used to supply the active agent 107. The vehicle gases can be selected to provide a desired ennment for a subject to which the active agent of the source 107 is supplied. For example, if the active agent is delivered to a patient as a breathable gas, the vehicle gases may include oxygen, nitrous oxide, or air in amounts sufficient to meet the needs of the patient. Other inert or active gases can be used.

In some embodiments, one of the source of gas 102 includes the source of active agent 107. The active agent of the source 107 can be a source of liquid gas that is vaporized by the vaporizer 108, or the active agent can be a source gaseous, such as a compressed gas under high pressure. The active agent can be mixed with one or more of the gas sources 102. The output controller 110 controls the amount of the gas mixture that is provided to the subject. The debugger 112 is a device or system that purifies and / or ventilates the gases that are provided to the subject. For example, if the active agent of the source 107 is provided as a breathable gas to a patient, the scrubber 112 can be used to remove the waste gases from the inhalant (such as the active agent), unused gas and exhaled carbon dioxide. . The aiarm / monitoring system 114 includes sensors that monitor the gas flow and / or gas content at one or more sites within the supply system 100. For example, the flow or amount of oxygen can be monitored when the active agent of the source 107 is provided as a breathable gas to a patient that ensures that the vehicle gases include sufficient oxygen for the patient. The alarm / monitoring system 114 also includes a user interface that is configured to provide a visual or audio alarm or monitoring of information to a user of the delivery system 100, such as a visual presentation, a light or audio alarm . The alarm / monitoring system 114 can be configured to notify the user when a predetermined condition is satisfied, and / or to provide information regarding the gas levels. With reference to Figure 10, a system 100A includes a high pressure circuit 116, a low pressure circuit 118, a breathing circuit 120 and a purification circuit 122. The high pressure circuit 116 includes the sources of compressed gas 102, which are connected to regulating valves 104b, 104a. The regulating valves 104a control the amount of gas flowing from each of the gas sources 102, and the regulating valves 104b can be opened to increase the gas pressure, for example, by providing an opening to the surrounding atmosphere. The low pressure circuit 118 includes the flow meters 106, the source of active agent 107 and the vaporizer 108. A gas mixture of the gas sources 102 is provided by the flow meters 106, which control the amount of each one of the gases from the gas source 102. As illustrated in Figure 10, the source of active agent 107 is a liquid. The source of active agent 107 is vaporized by the vaporizer 108, and added to the gas mixture. The breathing circuit 120 includes the output controller 110, two unidirectional valves 124, 126 and an absorber 128. The scrubber circuit 122 includes a valve 112a, a reservoir 112b and an output 112c. A subject 130 receives the gas mixture from the output controller 110, and the resulting gas is vented by the scrubber circuit 122. More specifically, the output controller 110 controls the amount of the gas mixture that is supplied to the subject 130 by unidirectional valve means 124. Expired gases flow through unidirectional valve 126 to valve 112a and reservoir 112b. The excess gas exits through the outlet 112c of the scrubber 112. Some of the gases can be recirculated and flow through the absorber 128 and in the breathing circuit 120. The absorber 128 can be a carbon dioxide absorber canister for the reduction of carbon dioxide gases from exhaled gases. In this configuration, the expired oxygen and / or the active agent can be recirculated and reused. One or more sensors S can be added in several positions in the system 100A. The S sensors detect and / or monitor the gases in the 100A system. For example, if one of the sources of gas 102 is oxygen, one of the sensors S may be an oxygen sensor configured and positioned to monitor the oxygen in the system 100A, so that the patient receives an adequate amount of oxygen. The sensors S are in communication with the alarm / monitoring system 114 (see figure 9). If undesirable or dangerous gas levels are present in the system 100, the alarm / monitoring system 114 can alert a user of the system 100A, so that an appropriate action can be taken, such as by increasing the oxygen levels administered to the subject. 130, or by disconnecting the subject 130 from the 100A supply system. Referring to Figure 11, there is shown a system 100B in which the source of active agent 107 is connected to two of the regulating valves 104b, 104a. If the source of active agent 107 is a source of liquid gas, an optional vaporizer 108 is provided to vaporize the source of liquid gas. If the source of active agent 107 is gaseous (e.g., a high pressure gas), then the vaporizer 108 can be omitted. The active agent of the source 107 is mixed with the other gas sources 102 in the low pressure circuit 118 in amounts that are controlled by the flow meters 106. The low pressure circuit 118 includes a gas reservoir 109 which contains any excessive flow of the gas mixture as it flows into the breathing circuit 120. It should be understood that the source of active agent 107 and / or any of the gas sources 102 , can be provided as a source of liquid gas with a vaporizer. System elements 100B illustrated in Figure 11 are essentially the same as those described above with respect to Figure 10, and will not be described in more detail. Methods of conformance with embodiments of the present invention which can be carried out using the systems 100, 100A, 100B, are illustrated in Figure 12. A mixture of one or more sources of respirable gas is provided (block 202). The sources of respirable gas can be obtained from the gas sources 102 as described with respect to Figures 9 to 11. A predetermined amount of the active agent is added to the gas mixture (block 204), as shown with respect to the source of active agent 107 in Figures 9 to 11. The gas mixture is administered to subject 120 (block 306). The exhaled gases are vented and / or recirculated (block 208), for example, by the scrubber 112. Although the methods of figure 12 are described with respect to the systems 100, 100A, 100B of figures 9 to 11, it is to be understood that any suitable system or device can be used to carry out the steps of Figure 12.. 2. Reduced Pressure Supply System Modes of a gas supply system 300 are illustrated with respect to Figure 13. The gas supply system 300 is positioned on a subject 302. The gas supply system 300 is particularly suitable for supplying an active agent in a gas mixture to the tissue of a subject 302, e.g., wound tissue. The system 300 includes a reduced pressure chamber 304 having a screen 306 that covers the treatment area of the subject 302. The reduced pressure chamber 304 is connected to a vacuum pump 310 via an outlet 310a of the pump. The reduced pressure chamber 304 includes an inlet 308a and an outlet 308b, which in turn are connected to a source of active agent 307. A controller 320 is connected to the active agent source 307 and the vacuum pump 310. Reduced pressure chambers and vacuum pump systems are discussed in US patents Nos. 5,645,081 and 5,636,643, the contents of which are hereby incorporated by reference in their entirety. The reduced pressure chamber 304 is configured to enclose an area of the subject 302 that provides a fluid-tight or gas-tight enclosure to effect treatment of the area with reduced or negative pressure, and the source of active agent 307. The pressure chamber 304 can be adhered to the subject 302 with a cover (not shown), such as a sheet of flexible, adhesive, and fluid impervious polymer. The cover may have an adhesive reinforcement that functions to cover the skin around the periphery of the area being treated, and to provide a generally gas-tight or fluid-tight seal and to keep the chamber 304 in place. The screen 306 is positioned over the treatment area of the subject 302. For example, if the treatment area of the subject 302 includes a wound, the screen 306 may be positioned over the wound to prevent its excessive growth. The size and configuration of the sieve 306 may be adjusted to suit the individual treatment area, and the sieve may be formed from a variety of porous materials. The material must be sufficiently porous to allow oxygen and other gases, such as gases from the source of active agent 307, to reach the treatment area. For example, screen 306 may be in the form of an open cell polymer foam, such as a polyurethane foam, which is sufficiently porous to allow gas to flow to and / or from the treatment area. Foams that vary in thickness and stiffness may be used, although it may be desirable to use a foamed material for patient comfort, if the patient must be on the apparatus during treatment. The foam may also be perforated to intensify gas flow and to reduce the weight of system 300. Screen 306 may be cut to a suitable shape and size to fit within the treatment area or, alternatively, screen 306 may be long enough to overlap with the surrounding skin. The vacuum pump 310 provides a suction source within the reduced pressure chamber 304. The source of active agent 307 provides an amount of the active agent to the reduced pressure chamber 304. The controller 320 controls the amount of vacuum applied to the reduced pressure chamber 304 by the vacuum pump 310, and the amount of the active agent that is supplied to the chamber 304 by the source of active agent 307. It should be understood that the controller 320 may apply a vacuum and / or the active agent in a substantially constant form, cyclically, or using various fluctuations or patterns, or any combination thereof. In some embodiments, the active agent is supplied by the active agent source 307 alternatively with the vacuum pump action of the vacuum pump 310. That is, the controller 320 alternately activates the vacuum pump 310, while deactivating the active agent source 307 and then activates the active agent source 307, while deactivating the vacuum pump 310. The pressure in the reduced pressure chamber 304 is allowed to fluctuate. In other embodiments, a substantially constant is maintained by vacuum pump 310, and active agent source 307 provides a substantially constant amount of active agent to chamber 304 in the reduced pressure environment. In some embodiments, a substantially constant pressure is maintained by the vacuum pump 310, and the amount of the active agent varies in a cyclic fashion. In other embodiments, the pressure in the reduced pressure chamber 304 is caused to fluctuate by the action of the vacuum pump 310, and the amount of active agent supplied by the source 307 also fluctuates. Fluctuations of the vacuum pump 310 and the resulting pressure in the chamber 304 or the amount of active agent supplied by the source 307 may be cyclical or non-cyclic The methods according to the embodiments of the present invention that can be carried out performed using the system 300, are illustrated in Figure 14. The chamber 304 is positioned over the treatment area of the subject 302 (block 402) The pressure is reduced in the chamber 304 by the vacuum pump 310 (block 404) A predetermined quantity of active agent from active agent source 307 is applied to the chamber (block 406). Although the methods of figure 6 are described with respect to system 300 of figures 5A-5C, it should be understood that any system or suitable device can be used to carry out the steps of figure 14. For example, the outlet 308b can be omitted, and the active agent can be supplied to the chamber 304 by the individual inlet 308a. also be added to the camera 304, for example, using a single input or an input and an output, as illustrated with respect to the active agent source 307 and the input 308a and the output 308b. In some embodiments, the vacuum pump 310 is attached to an additional collection container between the pump 310 and the chamber 304 for collecting exudates from the treatment area, for example, as described in U.S. Pat. No. 5,636,643. Negative pressure gas supply systems 300, as illustrated in Figure 13, are useful for the treatment of a variety of areas for treatment and, in particular, for the treatment of wounds. Wounds that can be treated using the 300 system include infected open wounds, decubitus ulcers, open incisions, partial thickness burns and various injuries to which flaps or grafts have been adhered. The treatment of a wound can be carried out by securing a gas supply system to the treatment site as shown and described previously, maintaining a substantially continuous or cyclic reduced pressure within the reduced pressure chamber 304, and supplying the active agent to chamber 304 in a substantially continuous or cyclic fashion, until the wound has reached a desired improved condition. A selected condition of improved condition may include sufficient granulation tissue formation for adhesion of a flap or graft, reduction of microbial infection in the wound, arrest or reversal of penetration of a burn, closure of the wound, integration of a flap or graft with the underlying wounded tissue, complete wound healing, or other stages of improvement or adequate healing for a particular wound type or wound complex. The gas supply system may be changed periodically, such as at 48-hour intervals, during treatment, particularly when using a gas supply system that incorporates a screen on or into the wound. The method can be implemented using a negative or reduced pressure ranging from 0.01 to 0.99 atmospheres, or the method can be implemented using a negative or reduced pressure ranging from 0.5 to 0.8 atmospheres. The period for the use of the method in a wound can be at least 12 hours, but can be extended, for example, by one or more days. There is no upper limit beyond which the use of the method ceases to be beneficial; the method can increase the closing speed until the time the wound actually closes. Successful treatment of various types of wounds can be achieved by using reduced pressures equivalent to about 5.08 cm to 17.78 cm of mercury under atmospheric pressure. The supply of reduced pressure to the gas supply system in an intermittent or cyclic fashion, as described above, may be useful for the treatment of wounds in the presence of the active agent. The intermittent or cyclic supply of reduced pressure to a gas supply system can be achieved by manual or automatic control of the vacuum system. A cyclic relationship, the ratio of time of "on" to "off" time in said treatment with intermittent reduced pressure, may be as low as 1: 10, or as high as 10: 1. A typical ratio is approximately 1: 1, which is usually achieved by alternating intervals of 5 minutes of supply and lack of reduced pressure supply. A suitable vacuum system includes any suction pump capable of providing at least 0.045 kg of suction to the wound, or up to 1,362 kg of suction, or up to 6,356 kg of suction. The pump can be any ordinary suction pump suitable for medical purposes, which is capable of providing the necessary suction. The dimension of the pipe that connects the pump with the reduced pressure apparatus is controlled by the capacity of the pump to provide the level of suction necessary for the operation. A 0.635 cm diameter tube may be suitable. The embodiments of the present invention also include methods of treating damaged tissue, including the steps of applying negative pressure to a wound and the active agent for a selected time and at a selected amount, sufficient to reduce the bacterial density in the wound. Open wounds are almost always contaminated with harmful bacteria. In general, a bacterial density of 105 bacterial organisms per gram of tissue is considered to produce infection. It is generally accepted that at this level of infection, the grafted tissue will not adhere to a wound. These bacteria must be destroyed, either through the natural immune response of the host or through some external method, before a wound closes. The application of negative pressure and active agent to a wound can reduce the bacterial density of the wound. It is thought that this effect may be due to the incompatibility of the bacteria with a negative pressure environment, or increased blood flow to the wound area in combination with exposure to the active agent, since the blood carries with it cells and enzymes that destroy the bacteria. The methods according to the embodiments of the present invention can be used to reduce the bacterial density in a wound by at least half. In some embodiments, they can be used to reduce the bacterial density by at least 1000 times, or by at least 1 million times. The embodiments of the present invention also include methods of treating a burn, including the steps of applying negative pressure and the active agent to the burn over an area with a predetermined reduced pressure and for a time sufficient to inhibit the formation of a burn. full thickness. A partial-thickness burn, one that has a surface layer of dead tissue and an underlying stasis zone, is often sufficiently infected, so that it will transform within 24 to 48 hours into a full-thickness burn, one in the which all the epidermal structures are destroyed. The application of negative pressure and an amount of the active agent to the wound can prevent the infection from becoming severe enough to cause the destruction of the underlying epidermal structures. The magnitude, pattern and duration of the application of pressure may vary with the individual wound. The embodiments of the present invention also include methods for enhancing the attachment of living tissue to a wound, comprising the steps of first attaching the living tissue to the wound to form a wound-tissue complex, then applying a negative or reduced pressure of selected magnitude and an amount of the active agent to the wound-tissue complex over a sufficient area, to promote the migration of subcutaneous tissues and epithelia to the complex, with negative pressure and exposure to the active agent being maintained for a selected period sufficient to facilitate wound closure. The union of living tissue to a wound is a common procedure that can take many forms. For example, a common technique is the use of a "flap", a technique in which tissue from the skin of an area adjacent to the wound is separated on three sides but remains attached in the room, and is then moved over the wound . Another technique that is often used is an open skin graft in which skin is completely separated from another surface of the skin, and grafted onto the wound. The application of negative pressure and active agent to the wound-graft complex reduces the bacterial density in the complex, and improves blood flow to the wound, thereby improving the binding of the grafted tissue.

E. Other Apparatus Within certain embodiments of the invention, it may be desirable to supplement the methods of the present invention for the treatment of patients who will be subjected to or have been subjected to trauma, with the ability to externally manipulate the temperature of the central part of the patient. Patient's body In this regard, the temperature of the central part of a patient's body can be manipulated, in combination with the methods of the present invention, by invasive or non-invasive means.

Invasive methods for manipulating the temperature of the central part of the body include, for example, the use of a cardiopulmonary pump that heats or cools the patient's blood, thereby raising or lowering the temperature of the central part of the patient's body . Non-invasive ways to manipulate the temperature of the central part of the body, include systems and devices that transfer heat into the patient's body, or outside it.

HIV Combination Therapies The compounds and methods of the present invention can be used in the context of many therapeutic and diagnostic applications. To increase the effectiveness of a treatment with the compositions of the present invention, such as oxygen antagonists, it may be desirable to combine these compositions with other effective agents in the treatment of those diseases and conditions (secondary therapy). For example, the treatment of stroke (anti-stroke treatment) typically includes an antiplatelet (aspirin, clopidogrel, dipyridamole, ticlopidine), an anticoagulant (heparin, warfarin) or a thrombolytic (tissue plasminogen activator). Various combinations can be used; for example, an oxygen antagonist, such as H2S, is "A", and secondary therapy is "B".

A B / A B / A / B B / B / A A / A / B A / B / B B / A / A A / B / B / B B / A / B / 'B B / B / 'B / A B / B / A / B A / A / B / B A / B / A / B A / B / B / A B / B / A A B / A / B / A B / A / A / B A / A / A / B B / A / A A A / B / A / A A A / B / A The administration of the oxygen antagonists of the present invention to biological material will follow general protocols for the administration of that particular secondary therapy, taking into account the toxicity, if any, of the treatment with the oxygen antagonist. It is expected that the treatment cycles are repeated as necessary. It is also contemplated that several standard therapies may be applied, as well as surgical intervention, in combination with the therapies described.

IX. Examples The following examples are included to demonstrate preferred embodiments of the invention. Those skilled in the art should appreciate that the techniques described in the following examples represent techniques discovered by the inventor that function well in the practice of the invention, and thus can be considered to constitute preferred modes for their practice. However, those skilled in the art should appreciate, in light of the present disclosure, that many changes can be made in the specific embodiments described, and that they can still achieve an equal or similar result without departing from the spirit and scope of the invention.

EXAMPLE 1 Preservation of nematodes in carbon monoxide The atmosphere contains 210,000 ppm of oxygen. Exposure to low levels of oxygen, or hypoxia, results in cell damage and death in humans. In the C. elegans nematode, oxygen concentrations between 100 ppm and 1000 ppm are also lethal. Through the critical study of the nematode response at a scale of oxygen tensions, it was found that oxygen concentrations below 10 ppm and above 5000 ppm are not lethal. In 10 ppm of oxygen balanced with nitrogen, the nematodes enter a state of reversible suspended animation in which all aspects of animation observed under the optical microscope cease (Padilla et al., 2002). At oxygen concentrations of 5000 ppm (balanced with nitrogen) and larger, nematodes progress through their life cycle normally. In a search for drugs that protect nematodes against hypoxic damage, carbon monoxide was tested. To achieve specific atmospheric conditions, the following apparatus was used: the barrel of a glass syringe that had a tip with an immobilization device such as a LUER-LOCK with the large opening of the barrel sealed with a steel and rubber accessory machined to the measure to achieve a hermetic seal, was secured by means of the immobilization device, to the inlet of an environmental chamber having an inlet and an outlet, each adapted with an immobilization device such as a LUER accessory -LOCK A defined gas was humidified and supplied to the ambient chamber, first venting the gas from a compressed tank (Byrne Specialty Gas, Seattle, WA) through a gas scrubber flask (500 ml, Kimex) filled with double distilled water. The gas washer bottle was connected to the environmental chamber beyond a gas flow meter. A gas flow meter was used to provide a regulated flow of 70 cc / min through the environmental chamber throughout the incubation for 24 hours. To test whether reversible stasis induced in C. elegans nematodes could be achieved, C. elegans embryos from 2 cells, L3 larvae or adult nematodes were collected and exposed to an environment of effectively 100% CO, an environment of 100 % of N2, an environment that comprised 500 ppm of oxygen balanced with carbon monoxide, or environments that comprised 100, 500 or 1000 ppm of oxygen balanced with nitrogen at room temperature. The nematodes were visualized using differential interference contrast microscopy (also known as Nomarski optics). Images were obtained, and analyzed using NIH image and Adobe Photoshop 5.5. The embryos were approximately 50 μm in length. The results of these experiments showed that 100% carbon monoxide was not lethal, and induced reversible suspended animation.

The nematodes did not survive 500 ppm of oxygen balanced with nitrogen; however, those treated with 500 ppm of oxygen balanced with carbon monoxide went into suspended animation and survived. See later.

EXAMPLE 2 Preservation of human skin in carbon monoxide Carbon monoxide is extraordinarily toxic to humans, because it competes strongly with oxygen for its binding to hemoglobin, the primary molecule that distributes oxygen to tissues. The fact that nematodes, which do not have hemoglobin, are resistant to carbon monoxide and still protected against hypoxic damage by this drug, suggested the possibility that carbon monoxide would protect against hypoxic damage in human tissue in situations where no blood is present, such as in blood-free surgical fields or tissue transplantation. To test this hypothesis, human skin was used. For this purpose, three human prepuce were obtained. The prepuce tissue was preserved in keratinocyte growth medium (KGM) containing insulin, EGF (0.1 ng / ml), hydrocortisone (0.5 mg / ml) and bovine pituitary extract (approximately 50 micrograms / ml protein). The foreskins were rinsed in PBS, and the excess fat tissue was removed. Each foreskin sample was divided into 2 equal pieces. Each piece was placed in a separate container containing a PBS solution with 24 mg / ml Dispase II (from EC 3.4.24.4 of Bacillus polymyxa: Roche Diagnostics Corp., Indianapolis, IN). A container (containing a piece of foreskin in PBS with Dispase II) was kept in a humid chamber in a ventilation hood. The other container (with the other half of the foreskin in PBS with Dispase II) was placed in the same ventilation hood in an environmental chamber perfused with 100% humidified CO. Both samples were kept at room temperature for 24 hours. The methods used to establish defined atmospheric conditions were identical to those used in Example 1. After 24 hours of exposure to normoxia or 100% CO, the keratinocytes were isolated from the foreskins according to the method described by Boyce et al. . (1983; 1985; citations incorporated herein by reference in their entirety). In summary, the epidermis of each foreskin sample was removed from a fresh plate containing PBS. The epidermis was comminuted and homogenized before incubation in 3 ml of trypsin at 0.05%, EDTA at 1 mM for 5 minutes, at room temperature, to separate the basal cells from the epidermis. After incubation, 6 ml of 400 μg / ml (micrograms per ml) of soybean trypsin inhibitor, 1 mg / ml of BSA was added, and the samples were centrifuged at 900 RPM. The supernatant of each sample was discarded, and the pellets of samples were resuspended in 10 ml of KGM. Each sample was divided into two 10 cm plates, each of which contained 5 ml of KGM and 100 μl of HEPES, pH 7.3 (N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid). Plates were incubated in a 37 ° C incubator perfused with 95% room air, 5% carbon dioxide, for five days. The cells were visually inspected using an inverted phase contrast microscope. Three of the keratinocyte populations exposed to normoxia showed little to no growth. Three of the keratinocyte populations exposed to 100% CO showed significant growth. The quantification of the number of viable keratinocytes, estimated by the formation of colonies, was performed for two of the three prepuces. See figure 1.

TABLE 1 Quantification of colony formation EXAMPLE 3 More information regarding example 1 The following example contains information that covers and extends the information described in Example 1.

A. Materials and methods Cameras and environmental devices. Oxygen deprivation experiments were carried out, using a custom made atmospheric chamber designed by W. Van Voorhies (Van Voorhies et al., 2000). The chamber is a 30 mL glass syringe (Fisher # 14-825-1 OB) adapted with. a custom-made steel cap that is lined with two Viton o-rings that ensure a hermetic seal. The plug is perforated through, and has a steel Luer Lock on the outer face, so that a hose carrying compressed gas can be fixed. A defined gas mixture is supplied to the chamber at a constant pressure and flow rate from compressed tanks by first passing through a rotometer (Aalborg, flow tube number 032-41 ST) or mass flow controller (Sierra Instruments # 810) that monitors the flow rate, and then through a 500 ml gas scrubber bottle (Fisher # K28200-5001) containing 250 ml of water to hydrate the gas. Nylon tubing with an outer diameter of 0.635 cm (Cole-Parmer # P-06489-06) or FEP (Cole-Parmer # A-06450-05) was used and connections were made between the pipe and the regulators and between the pipeline and the pipeline. Rotometers with brass John-Guest type accessories (Byrne Gas). All other connections were made with Microflow quick connect fittings (Cole-Parmer # A-06363-57, # A-06363-52) or standard Luer accessories (Cole-Parmer # A-06359-37, # A-06359- 17). Viability of nematodes in hypoxia. The N2 Bristol strain was continuously maintained at 20CC, taking care to ensure that the population did not die of starvation. Adult C. elegans of logarithmic phase were selected in a drop of sterile water containing 100 μg / ml of amplilin, 15 μg / ml of tetracycline and 200 μg / ml of streptomycin on a glass plate. Adults were bitten with a shaving blade, and 2-cell embryos were selected using a pipette. 30 to 60 embryos from 2 cells were transferred to a small glass pan (custom made to adjust atmospheric chambers, Avalon Glass Works, Seattle WA) filled with 3 ml of 1% agarose in M9. The troughs were then placed in a humid chamber for 2 hours, and the embryos were allowed to reach maturity, and were then placed in the environmental chamber. The environmental chambers were perfused continuously at room temperature with pure N2 (grade 4.5), 100 ppm 02 / N2, 500 ppm 02 / N2, 1000 ppm 02 / N2 or 5000 ppm 02 / N2 at 70 cc / min 24 hours. After the exposure, pieces of agarose containing the embryos were cut from the trough and placed with the embryos facing up on a NGM plate sized with medium seeded with E. coli (OP50). The embryos were scored for hatching 24 hours after exposure, and the hatched L1 steps were transferred to the surface of the NGM plate, and the course of the plates was followed until the age of admission. The animals that could not be given were subtracted from the total. All gases were provided by Byrne Gas (Seattle, WA). It was ensured that pure N2 contained less than 10 ppm of impurities, and all 02 / N2 mixtures were certified at + 2% oxygen content (for example, it was certified that 100 ppm of 02 / N2 contained between 98 ppm of 02 and 102 ppm of 02). The conversion of parts per million to kPa was based on 1 million parts = 101 kPa at 1 atmosphere. Feasibility of nematodes in atmospheres based on carbon monoxide. 30 to 60 embryos of Bristol N2 and hif-2 (ia04) strains kept continuously were harvested as described above. The environmental chambers were perfused continuously at room temperature with pure CO (CP grade) or 500 ppm 02 / CO at 70 cc / min for 24 hours. To achieve 2500 ppm 02 / CO or 2500 ppm 02 / N2, 5000 ppm 02 / N2 was mixed at a 1: 1 ratio with pure CO or pure N2 using two mass flow controllers (Sierra Instruments 810) that would monitor the flow with precision. Each gas was supplied in a 3-way valve (Cole-Parmer # A-30600-23) at 50 cc / min, and the resulting mixture was then passed through a gas washer bottle and an environmental chamber throughout of the exhibition for 24 hours. All gases were provided by Byrne Gas (Seattle, WA). The 500 ppm 02 / CO mixture was certified to ± 2% of the oxygen content, and contained 7000 ppm of N2 to ensure a consistent ratio of 02 / CO during use of the tank. Biological analysis of cells. To determine the degree of progression of development in nitrogen-based atmospheres (Table 2), embryos from 2 cells were exposed to various degrees of hypoxia as described above, and were photographed immediately, or photographed after a recovery period of 12 hours in a humid chamber. To determine if the embryos stopped their development in carbon monoxide-based atmospheres, 2-cell embryos were brought to maturity in ambient air for two hours, and photographed immediately, or put in 100% carbon monoxide or 0.05 kPa of 02 / CO for 24 hours, and were photographed immediately after exposure. In all cases, DIC microscopy was performed by placing the embryos under a coverslip on a thin pad of 1% agarose, and observation in a Zeiss axioscope. Photographs were taken using RS image and Adobe Photoshop software.

B. Results It has been previously reported that HIF-1 is required in C. elegans in moderate hypoxia (0.5 kPa of 02 (Padilla et al., 2002) and 1 kPa of 02 (Jiang et al., 2001)), and he knows that animation suspended in anoxia (> 0.001 kPa of 02) is possible (Padilla et al., 2002). To accurately define the scales in which each of these responses are active, the viability of wild-type C. elegans embryos was determined after exposure to various oxygen tensions between moderate hypoxia and anoxia for 24 hours. The embryos exposed to anoxia entered suspended animation as previously reported, and in this way they survived the exposure with high viability. The embryos in 0.5 kPa of 02 continued to be in suspended animation throughout the exhibition, and they also survived with high viability. However, the embryos exposed to an intermediate scale of oxygen tensions between moderate hypoxia and anoxia (0.1 kPa from 02 to 0.01 kPa from 02), surprisingly did not survive (Fig. 2). The embryos did not hatch during exposure to this intermediate scale of hypoxia, indicating that they did not successfully execute the HIF-1 mediated response. To determine if they appeared to be in suspended animation, it was examined whether embryos in this intermediate scale stopped their embryogenesis during exposure. The embryos in lethal oxygen tensions did not interrupt their embryogenesis, and the increased amounts of oxygen correlated with an increase in the degree of progression of development in the embryo (Table 2). After reoxygenation, most of these embryos failed to hatch, and many of those that hatched stopped as abnormal L1s. These data show that this intermediate scale of hypoxia is a unique tension in which oxygen levels are neither high enough to facilitate continuous animation, nor are they sufficiently low to induce suspended animation. Based on these findings, it was hypothesized that if carbon monoxide, a competitive inhibitor of oxygen binding, could induce suspended animation in the presence of low oxygen levels, it would provide protection against this lethal scale of hypoxia. To examine this possibility, the viability of C. elegans embryos at various concentrations of carbon monoxide was first determined. Despite the toxic effects that high levels of carbon monoxide can have on some systems, it was found that C. elegans embryos are remarkably tolerant of a wide range of carbon monoxide stresses. In fact, C. elegans embryos can withstand continuous exposure to 101 kPa of CO (100% CO) for 24 hours with high viability (81.5% survival to adulthood, Figure 3). Notably, at 101 kPa of CO, embryos did not progress through embryogenesis during exposure, indicating that they entered suspended animation. To test if carbon monoxide could protect embryos in the presence of lethal oxygen tensions, the viability of embryos exposed to 0.05 kPa of 02 balanced with carbon monoxide was determined. In contrast to embryos exposed to 0.05 kPa of 02 balanced with N2 (most of which do not survive), these embryos recovered with 96.2% viability until adulthood (figure 3). In addition, like embryos treated with 101 kPa of CO, embryos in 0.05 kPa of 02 balanced with carbon monoxide stopped their embryogenesis, indicating that they entered into suspended animation. Therefore, carbon monoxide can protect against hypoxic damage in the presence of lethal oxygen tensions, inducing suspended animation.

To better examine the scale of oxygen tensions that can be protected by excess carbon monoxide, embryos that lacked the function of HIF-1 (the strain hif-1 (ia04)) were used to record whether the protection against damage Hypoxic was also possible in moderate hlpoxia. After carrying out tests of several oxygen voltages between 0.1 kPa of 02 and 1 kPa of 02 balanced with nitrogen, it was found that the maximum requirement for HIF-1 was 0.25 kPa of 02 balanced with nitrogen. In this atmosphere, wild-type embryos progress normally through development and exhibit high viability, but hif-1 (ia04) embryos do not complete embryogenesis, and exhibit 100% lethality (Table 3). Therefore, it was examined whether carbon monoxide could protect embryos hif-1 (ia04) at 0.25 kPa from 02. At 0.25 kPa from 02 balanced with carbon monoxide, wild-type embryos and hif-1 (ia04) they entered suspended animation, and survived exposure with high viabilities (78.7% and 84.0% survival to adulthood, respectively) (Table 3). In this way, the induction of animation suspended by carbon monoxide is possible at oxygen tensions as high as 0.25 kPa of 02, and carbon monoxide may provide protection against moderate hypoxia, even in the absence of the HIF- function. 1.

TABLE 2 Quantification of the progression of development in hypoxia Wild-type embryos of 2 cells were placed in various degrees of hypoxia for 24 hours, and scored to the extent to which they progressed through embryogenesis. Exposure to atmospheres containing increased amounts of oxygen resulted in increased progression through embryogenesis. The percentage of embryos that stopped their development within a determined embryogenesis scale of 20 to 40 minutes was determined. The data is the result of 3 independent experiments.

TABLE 3 Carbon monoxide protects hif-1 embryos against ade- quate hypoxia ' Viabilities were tested to adulthood after 24-hour exposure of 0.25 kPa of 02 / N2 or 0.25 kPa of 02 / CO in wild-type embryos and hif-1 (ia04). All data points are the result of at least 3 independent experiments, and the values that could not be given were subtracted from the total.

Viability of nematodes in response to hypothermia Viability of nematodes is also sensitive to temperature, with 100% of a population dying after exposure for 24 hours at low temperature (4 ° C, Figure 15). However, if stasis is induced in the nematodes by equilibrium under anoxic conditions (<10 ppm oxygen) for 1 hour before the temperature decrease, a substantial proportion of them survive after a 24 hour exposure to 4 hours. ° C (figure 15). In this experiment, the nematodes were kept in stasis during the hypothermia period, and for 1 hour after they had been returned to room temperature. The anoxic conditions (pure N2), growth conditions and viability measurements are as described below.

EXAMPLE 4 Reduction of the temperature of the central part of the body and respiration in mice A. Materials and methods Implantation of telemetry devices. Female C57BL / 6J mice (Jackson Laboratories - Bar Harbor, Maine) were implanted with telemetry devices (PDT-4000 HR E-Mitter-MiniMitter Inc. - Bend, OR), according to the standard protocol provided by the manufacturer. The mice were allowed to recover for several weeks to allow the heart rate and body temperature signals to stabilize. The temperature of the central part of the body, heart rate and movement of the mice, were monitored continuously by means of the telemetry devices, and were recorded using the VitalView software (provided by MiniMitter). The room temperature was monitored using a HOBO (Onset Computer Corp. - Pocasset, MA), and the data was analyzed using the BoxCar software (provided by Onset Computer Corp.). Exposure of the mice to a regulated atmosphere. Each mouse was exposed to 1 L / min of (a) an atmosphere containing 500 ppm of H2S balanced with nitrogen (Byrne Specialty Gas - Seattle, Washington) mixed with ambient air (using a 3-channel gas proportioning meter, Aalborg - Orangeburg, New York), to give a final concentration of 80 ppm of H2S and 17% of 02, or (b) a nitrogen atmosphere mixed with ambient air to give a final concentration of 17% of 02. Measurements were made of H2S and 02 using an Innova GasTech GT series portable gas monitor (Thermo Gas Tech - Newark, California). Before and during exposure to testing in regulated and unregulated atmospheres, the mice were placed in a gasification chamber that included a glass cage (with drinking water and no food) adapted with inlet and outlet tubes FEP tubing from Cole-Parmer (Vernon Hílls, Illinois) for introduction and ventilation of the atmosphere. The cage was sealed with a lid using Dow Corning vacuum silicone grease (Sigma - St. Louis Missouri). The gas in each cage was vented through the discharge tube in the chemical hood. To ensure that the system is gas tight, a GasTech GT portable monitor was used to detect leaks. Respirometry In some experiments, oxygen consumption was measured by the use of a 02 PA-10a analyzer (Sable Systems), which was used according to the manufacturer's instructions. In addition, the carbon dioxide that was being produced by the animals was monosurveled using a C02 / H20 LI-7000 analyzer (Li-Cor Company) according to the manufacturer's instructions. These instruments were put in line with the environmental chambers, so that they would take samples from the gas inlet and discharge pipe. Regulation of the room temperature. The mice were housed in a Shel Lab low temperature daylight incubator (Sheldon Manufacturing Inc. - Comelius, Oregon), to regulate the temperature and light cycle (lights on at 8 AM, lights off at 8 PM. ) for mice. The mice were exposed to a regulated atmosphere as described above. When the mice were exposed to the regulated atmosphere, the temperature inside the incubator decreased to the desired temperature, for example, up to 10 ° C or 15 ° C. The mice were kept in the regulated atmosphere and at the lowered temperature for six hours. The atmosphere in the gasification chamber was replaced with ambient air, and the mice were returned to normal room temperature (22 ° C), and allowed to recover.

B. Results Reference line data. To determine the response of the mice to sublethal doses of hydrogen sulfide, the inventor first established baselines for core temperature, heart rate and movement, recording data over a period of one week of four mice with implanted transceivers, in the incubator maintained at room temperature and perfused with room air. The baseline data showed that the mice have a circadian rhythm with maximum activity at night right after the lights go out, and in the early morning hours just before the lights are turned on. The temperature of the central part varied from a high value of 37 ° C during its active periods, to a low value of 33.5 ° C during its inactive periods. The heart rate varied from 750 bpm (beats per minute) during their active periods, to 250 bpm during their inactive periods. It is likely that the heart rate correlates with the temperature of the central part (higher temperature, higher heart rate). Also, the obvious motor movement was higher during the evening and shortly before dawn. Exposure of mice to atmospheres regulated at room temperature. The first test of the exposure of a mouse to hydrogen sulfide, included putting the mouse first in the gasification chamber maintained at 27 ° C in the incubator for one hour. After the hour, the chamber was perfused with 80 ppm as described above in general, and the temperature of the incubator was reduced to 18 ° C throughout the duration of the experiment. Although no immediate changes in heart rate and obvious motor movement were detected, a dramatic decrease in core temperature was observed. The experiment was allowed to proceed for 90 minutes, during which time the core temperature decreased to 28.6 ° C - five degrees below the lowest record for any of the four mice in the baseline study described above. During the recovery after the chamber was perfused with ambient air, the inventor noticed that the animal was initially relatively immobile (it was easy to catch); however, within 60 minutes, he had returned to a normal scale of activity and temperature of the central part. A second mouse was exposed to the same protocol; however, this time, gasification at 80 ppm was carried out for 3 hours. During this time, the inventor noticed that the heart rate decreased significantly from 600 bpm to 250 bpm, the obvious motor movement showed almost no activity, and the temperature of the central part decreased to 18.6 ° C. Changes in respiration accompany the decrease in temperature of the central part. The exposure of the mice to 80 ppm of H2S also results in a decreased metabolic rate, as determined by measuring oxygen consumption and carbon dioxide production. For example, a mouse in which the temperature of the central part and the production of carbon dioxide had been simultaneously measured, demonstrated a rapid decrease in carbon dioxide production preceding the decrease in temperature of the central part of the animal ( 4A). The approximately three-fold reduction in carbon dioxide production established a new baseline in approximately 5 minutes after exposure to H2S. Table 4 shows the results of an experiment with concurrent measurements of O2 and C02 concentrations of mice exposed to ambient air from which CO2 impurities were extracted (hence, values of 0 for controls), with or without H2S (80 ppm). The measurements were made over a period of 15 minutes, keeping the mice in a sealed environmental chamber of 0.5 L with flow rates of 500 cc / min. Oxygen consumption is obtained by subtracting the oxygen concentration when the mouse is present, of control when the mouse is absent. Also, the production of carbon dioxide is obtained by subtracting the concentration of carbon dioxide when the mouse is present, from the control when the mouse is absent. RQ represents the respiratory quotient, and is equal to the ratio of carbon dioxide produced to produced oxygen. This result shows a 2 to 3-fold decrease in oxygen consumption in the presence of H2S, as well as a 3 to 4-fold decrease in carbon dioxide production. The change in respiratory quotient reflects the disparity of oxygen consumption and carbon dioxide production by the mice in the presence or absence of H2S.

TABLE 4 Exposure to H2S inhibits respiration in mice The different stasis parameters (reduction in oxygen consumption, decrease in carbon dioxide production or decrease in mobility) can be tested by a variety of tests and techniques. For example, probably the easiest way to measure the induction of stasis in mice that are administered H2S, is through the observation of their respiration. Of course, this covers all three parameters, because it is indicative of decreased oxygen consumption, carbon dioxide production and mobility. A normal mouse in ambient air under standard conditions will have approximately 200 breaths per minute. If H2S is administered to the mouse at 80 ppm, and the temperature of the central part is reduced to 15 ° C, the respiration decreases by at least an order of magnitude between 1 to 10 breaths per minute. In fact, a mouse was observed under these conditions that did not have a breath for a period greater than one hour, indicating that deep levels of stasis are achieved. In this way, this represents a decrease of at least about 1 to 20 times in cellular respiration (i.e., oxygen consumption and carbon dioxide production). Exposure of mice to regulated atmospheres at low ambient temperatures. To begin to define the limits of the capacity of hydrogen sulfide to reduce the activity in mice, the inventor carried out several experiments in which a mouse without telemetry was used, followed by exposure of a mouse that has telemetry to acquire the data . The first experiment was to subject a mouse without telemetry to a controlled atmosphere of H2S at 80 ppm at a reduced cabinet temperature of 10 ° C essentially as described in the materials and methods section as above, except that the mouse put in the gasification chamber for one hour at 27 ° C before exposure to the gas and reduction in room temperature. The mouse without telemetry responded well to this treatment, and recovered its activity within approximately 90 minutes after the removal of the gasification chamber. The telemetry mouse that was subjected to the same conditions also responded well to treatment, and showed a decrease in core temperature to approximately 12.5 ° C. The Inventor could not accurately determine this temperature, because the electronics failed at 15.3 ° C. The decrease in temperature to 12.5 ° C is therefore an estimate based on the slope of the decrease before the fault and the time the animal remained in the chamber after the failure of the electronics. Due to the limitation of the equipment, the Inventor then tested each of the four mice with telemetry for a period of 6 hours in the gasification chamber with a regulated atmosphere containing approximately 80 ppm of hydrogen sulfide, or with ambient air essentially as described above. The temperature of the incubator was reduced at the beginning of the experiment (exposure to the regulated atmosphere, or time 0 for mice exposed to ambient air) up to a constant temperature of 15 ° C. At the end of the 6 hour period, the mice were returned to an atmosphere of ambient air and an ambient temperature of 22 ° C as described above in general. There was a clear decrease in the temperature of the central part of the body in the four mice, which depended on the use of 80 ppm of hydrogen sulfide (Figure 4B). There was also a noticeable decrease in heart rate and obvious motor movement associated with the decrease in temperature. The mice were maintained for 4 weeks without apparent change in the behavior of the animals.

EXAMPLE 5 Studies in murines on the reduction of radiation injury A. Scientific rationale Although aspects of the radiation injury model can be evaluated and evaluated in cell culture, to test the capacity of an experimental drug to affect the injury and the healing process, requires the inclusion of all the response systems that are affected. At this point in time, the only way to achieve this is in an entire animal. The inventor is proposing the use of mice for such studies as the most appropriate model. C57BL / 6 mice have been selected for study because this strain of mouse is easily susceptible to lung injury by radiation, the level of radiation that is tolerated in this strain has been established, and the inventor has recently shown that H2S the temperature of the central part of this strain of mice decreases. Two identical experiments are planned under this protocol. Each experiment will investigate the efficacy of H2S-induced hypothermia on the development of radiation-induced lung injury. Ten mice per group will be exposed to one of four test conditions (H2S / chest irradiation of 17.5 Gy, H2S / without chest irradiation, without H2S / thoracic irradiation of 17.5 Gy, or without H2S / without thoracic irradiation), followed then for 13 weeks. Twelve animals per group will also be exposed and followed for 26 weeks (the increased n is required to compensate for the increased mortality that occurs later in the course of the disease). For these experiments, analysis of variance (ANOVA) will be used as the statistical model for data analysis. A fully crossed and randomized two-way ANOVA with 4 groups (irradiated or non-irradiated mice receiving HS or not receiving H2S) and two time intervals (13 or 26 weeks) will be used to analyze temporary changes in the number of cells inflammatory effects of bronchoalveolar lavage and total protein concentration and levels of hydroxyproline in the lung. Assuming 80% power, 5% significance and a two-tailed test, five surviving mice per group combination with injury, Intervention group and time point, will allow a detectable difference between group means greater than or equal to 1.7. times the underlying standard deviation within the group. The standard deviation within the group is expected to be approximately 25%. In this way, changes in the number of inflammatory cells or collagen content in the lung of 35 to 50% of the control values should be discernible in these experiments. Exposure to H2S and thoracic irradiation will be done in SLU AHR in a linear accelerator set. Bronchoalveolar lavage and lung collection at necropsy will be performed at the necropsy site of AHR mice. The content of bronchoalveolar lavage cells and measurements of hydroxyproline content in lungs and protein concentration will be carried out in the other laboratory (D3-255). Wild genotype C57BL / 6 mice will receive 17.5 Gy of thoracic irradiation. Mice will be anesthetized with intraperitoneal Avertin, placed in tissue closures for individual mice, and irradiated by the linear accelerator with 8.5 Gy at a dose rate of 3 Gy / min through two collimated side fields to choose as target only the thorax (total thoracic dose of 17.5 Gy).

B. Protocol Anesthesia. Wild-type C57BL / 6 mice will be anesthetized for intratracheal dosing with isoflurane. The depth of anesthesia will be monitored by respiratory rate to respond to tactile stimulation. Intraperitoneal injection of Avertin (0.4-0.7 ml / mouse, i.p.) will be used to anesthetize the animals for the thoracic irradiation procedure. The depth of anesthesia will be monitored by respiratory rate and response to tactile stimulation. Exposure to hydrogen sulfide. The mice will be placed in a closed Plexiglas gasification chamber similar to that previously used for mice (IR1606). The camera will have two holes (entry and exit). A gas containing H2S (80 ppm) balanced with ambient air will be vented through the chamber at a rate of 1 liter per minute. The site gas will be vented using the internal ventilation system with a hose that extends from the exhaust vent to the suction vent for the site. Administration of dangerous agents. The mice will be irradiated while in the gasification chamber with a total dose of 17.5 Gray using the linear accelerator. This dose of radiation will induce a subacute lung injury in mice, which progresses to fibrosis. The mice will not be radioactive or otherwise pose a risk to personnel or other animals. No monitoring, containment or special provision is required due to irradiation. Projected euthanasia. At almost weeks 13 and 26 after the thoracic irradiation, the animals will be euthanized by means of deep anesthesia (using Avertin 0.4-0.7 ml, i.p.), followed by exsanguination by puncture of the inferior vena cava. Bronchoalveolar lavage will be performed to determine the number of inflammatory cells, differential counts and protein concentration of the lavage fluid. Lung and esophagus tissue will be removed for histological evaluation and analysis of collagen content. Dying animals. Thoracic radiation is associated with a finite mortality rate in mice, with 15% dying around week 10, and 50% around week 22 post-irradiation. Researchers will monitor the animals daily for adverse effects (2 to 3 times per day initially, until they appear to be stable, then once a day until the disease begins to progress, at which point the inventor will make daily observations again multiple). If an animal is losing weight, can not groom itself and exhibits severe respiratory distress and / or clumsy or significantly decreased movement, it will be euthanized with Avertin overdose. When practical, bronchoalveolar lavage and tissue collection for histology will be performed for these non-projected euthanasia. The thoracic irradiation must produce a lesion of the lung that by itself is not painful, but may manifest itself (week 10), through increased respiratory rate, moderate loss of appetite, moderate weight loss and / or inability to groom. Researchers and the personnel in charge of the animals will monitor the animals daily for such adverse effects. If an animal does not appear to be eating, it will provide soft feed and fluid support. If the animal is perceived to be in pain, analgesia with butorphanol (0.2 mg / kg i.p.) or buprenorphine (1.0 mg / kg twice daily the next day) will be administered as needed. If an animal seems to be suffering and palliative measures do not lead to any improvement, it will be euthanized immediately. Lung and esophageal tissue will be collected for histopathological evaluation and analysis of the collagen content in the projected necropsies. Post-irradiation management. To minimize the risk of transmitting pathogens to the rest of the facility, and to protect these animals while they are a little immunocompromised, all the management work on these animals will be done first thing every day (before any other animal in the Facility), and it will be done in a biosecurity cabinet. To minimize the risk of secondary infections, the mice will have cages and bed of straw treated with an autoclave. In addition, they will be supplied with standard food for rodents that has been irradiated to destroy the pathogens. Wild genotype C57BL / 6 mice will receive 17.5 Gy of thoracic irradiation. The mice will be anesthetized with intraperitoneal Avertin, placed in individual cloth rabbits, and moved to them in a closed plexiglass gasification chamber similar to that previously used for mice (IR1606). The camera will have two holes (entry and exit). A gas containing H2S (80 ppm) balanced with ambient air will be vented through the chamber at a rate of 1 liter per minute. The site gas will be vented using the internal ventilation system with a hose that extends from the exhaust vent to the suction vent for the site. Once in the gassing chamber, the mice will be irradiated by the linear accelerator with 8.5 Gy at a dose rate of 3 Gy / min through two collimated side fields to target only the thorax (total chest dose). 17.5 Gy). After concluding the thoracic irradiation, the animals will be returned to their monitored micro-isolating cages until they recover from the anesthesia.

Necropsies projected. A group of animals will undergo necropsy at week 13 post-irradiation, to evaluate the inflammatory phase of the lesion. The second group will be euthanized in week 26 to evaluate the fibrotic phase of the lesion. The animals will be anesthetized with Avertin, and then they will be subjected to exsanguination. The lungs will be washed with 1000 μl of PBS, and the washing fluid will be kept on ice for total and differential cell count. The right lung will then be harvested for hydroxyproline content, and the left lung will be infused with 10% NBF at 25 to 30 cm of pressure through the trachea. The esophagus, trachea, left lung and heart will be immersed in NBF at 10%, and will be sent to the FHCRC histology shared resource laboratory for processing and pathological evaluation. The thoracic irradiation must produce a lesion of the lung that by itself is not painful, but may manifest itself (week 10), through increased respiratory rate, moderate loss of appetite, moderate weight loss and / or inability to groom. Researchers and the personnel in charge of the animals will monitor the animals daily for such adverse effects. If an animal does not appear to be eating, it will provide soft feed and fluid support. If it is perceived that the animal suffers pain, analgesia with butorphanol (0.2 mg / kg i.p.) or buprenorphine- (1.0 mg / kg twice daily the next day) will be administered as necessary. If an animal appears to be suffering and palliative measures do not lead to any improvement, it will be euthanized immediately by asphyxia with C02. It is likely that the primary problems are esophagitis (resulting in decreased consumption of water and food) and respiratory failure (reducing oxygen uptake). The inventor will be checking these animals 2 to 3 times a day until he is convinced that they are stable and respond well to treatment, at which point the inventor can reduce the frequency of observations up to once a day, until the disease begin to progress, at which point you will return to make multiple daily observations. Supportive care will be provided in several ways. If an animal is not eating or drinking well (evidence of weight loss and grooming problems), the inventor will provide soft feed and attempt fluid supplementation (lactated Ringer's solution, 1 to 2 ml per mouse, subcutaneously, using a needle of small hole (> 20 G), 1-2 times a day). If the animal is perceived to be in pain, analgesia with butorphanol (0.2 mg / kg i.p.) or buprenorphine (1.0 mg / kg twice daily the next day) will be administered as needed. If an animal appears to be suffering and palliative measures do not lead to any improvement, it will be subjected to immediate euthanasia by asphyxia with C02. In case an animal experiences significant pain or suffering at the time of the thoracic irradiation, the animal will be euthanized by asphyxia with C02. A third experiment consisted in subjecting a mouse with telemetry to a controlled atmosphere of H2S at 80 ppm at a reduced cabinet temperature of 10.5 ° C, essentially as described above.

During the experiment, the mouse was observed visually, and their movements were recorded by webcam, and telemetry measurements were recorded as described above. The mouse was exposed to a regulated atmosphere of 80 ppm H2S, and the cabinet temperature was reduced to a constant temperature of 10.5 ° C. At the end of a period of approximately 6 hours, heat was applied to the cabinet by adjusting the cabinet temperature to 25 ° C. The mouse was allowed to warm in the regulated H2S atmosphere until the core temperature of the mouse was between 17 ° C and 18 ° C, time after which the regulated atmosphere was replaced with ambient air. There was a clear decrease in the temperature of the central part of the mouse body to 10.5 ° C in the regulated atmosphere, accompanied by a noticeable decrease in obvious motor movement. The respiratory rate decreased to an undetectable value by visual observation for approximately one hour and fifteen minutes. After the cabinet was heated, weak breathing was observed when the temperature of the central part of the mouse body reached 14 ° C. During the warm-up phase, when the temperature of the central part of the body rose to between 17 ° C and 18 ° C and the mouse was exhibiting breathing and movement, the regulated temperature was replaced with ambient air. Normal movement and breathing were completely evident when the temperature of the central part of the body returned to 25 ° C. The mouse exhibited no apparent change in its behavior, compared to animals that were not treated.

EXAMPLE 6 Studies in cells and mammals A. Studies in canines Studies were carried out on canines with dogs surgically implanted with telemetry devices to monitor the temperature of the central part of their body. The animals will be studied in the presence or absence of a sub-lethal dose of hydrogen sulphide for 10 hours. During this time, they will be continuously monitored for vital signs by telemetry. The ambient temperature will also be reduced to 15 ° C for 30 minutes, to determine if this has any effect on the temperature of the central body part of the animals. The procedure will be done with 2 groups of 2 dogs (four in total). Due to the cost of telemetry equipment, the inventor will do these experiments in succession. If the results of the first group indicate that the hypothesis is incorrect, the study will be repeated with the second group of two dogs. If the results of the second group do not support the hypothesis, the project will be discontinued. Toxicology studies show that, while the level of H2S is above the limit of OSHA for humans (10 ppm), it has previously been shown that the exposure of rats and mice to 80 ppm of H2S for 6 hours per day, 5 days per week, for 90 days, showed no adverse effect observed. This included obvious and histopathological recognition of the bowel, lung, heart, liver, kidneys or other organs, carried out at the end of the treatment. For the knowledge of the inventor, no information is available regarding the exposure of dogs to hydrogen sulfide. A critical problem when working with H2S, will not exceed the dose (80 ppm) described by others who have published studies on rodents exposed to hydrogen sulfide and have not observed harmful effects. There is considerable experience in the available anesthetic gas sciences, and the inventor is capable of delivering the gas to the mice at the prescribed dose. Many precautions are taken to ensure that animals and researchers are not harmed. These precautions include constant monitoring of the gas mixture with alarm adjusted to OSHA limits and sensitivity to 1 ppm, and a variety of equipment that is capable of mixing and delivering gas according to specifications, without any leakage inside and outside the system. A timeline for the protocol is given in table 5.

TABLE 5 Study timeline Day Activity Detail A CBC / chemistry will be done; he will undergo fasting -1 P re-surgery to the dog in the afternoon, but you will be left free access to water. Transdermal fentanyl patch placed in the afternoon of the day before surgery for preventive anesthesia. Preoperative placement of cephalic catheter; premedication with acepromazine, buprenorphine, glycopyrrolate; induction with ketamine: diazepam or propofol to allow intubation; maintenance of anesthesia by isoflurane and oxygen. The dog will be placed on dorsal inclination, and his abdomen will be stapled / prepared and wrapped in a field. Pulse monotoring, respiratory rate, carbon dioxide of final tidal volume, inhaled percentage of anesthetic agent, SpO2, will be performed and recorded every 15 minutes or more frequently. Fluid support will occur during and after surgery. Once the dog is stable and prepared Surgery for the procedure, a laparotomy will be performed in the ventral midline, beginning caudal to the umbilicus and extending caudally 5 to 10 cm. A sterile transmitter will be placed in the peritoneal cavity. The placement will be monitored to ensure that the transmitter is able to move freely; the omentum will be replaced, and the peritoneal cavity will be closed in three layers. The dog will be monitored until it is extubated, is able to show thermoregulation, and is steeply inclined. Daily monitoring of the incision site, abdomen (by means of palpation and ultrasound, if indicated), appetite, temperature (for the first 3 to 5 days post-operative), weight and activity of the dog will be performed.

TABLE 5 (CONTINUED) B. Human Plaguetas To test the concept that the use of inhibitors of oxidative phosphorylation could be used for human benefit, the inventor induced a state of suspended animation in human tissues to protect them from lethal exposure to oxygen. In pilot experiments, the inventor put human skin in an environment of 100% CO. The inventor notes that after 24 hours, the skin cells survive 100 times better in CO than in ambient air. These results are very exciting; provide evidence that inhibitors of oxidative phosphorylation can be effective in human tissues. Another series of experiments demonstrates the protective effects of suspended animation induced on platelets. One unit of platelets was divided in half. The first half was maintained under standard storage conditions, which include keeping the platelets at room temperature (22-25 ° C) with constant agitation. The other half was placed in an anoxic environment (<10 ppm oxygen) using standard methods to remove oxygen. The two series of platelets were compared on days 0, 5 and 8. Platelets maintained under anoxic conditions responded well or better than those maintained under standard conditions on a panel of five different in vitro tests, including the ability to aggregate, cell morphology , fining with annexin-V (phosphatidyl serine backslide towards the outer membrane as an initial apoptotic marker), etc. This indicates that the control of metabolic activity, specifically oxidative phosphorylation, can be achieved by the removal of oxygen, and has a protective effect on cell function during long periods of stasis. Hydrogen sulfide is able to bind to cytochrome c oxidase, as well as to CO, and stop oxidative phosphorylation upon request. It is so powerful to prevent oxidative phosphorylation, that if a person takes a single breath in an atmosphere with 0.1% hydrogen sulfide, he will not take one more. Rather, it collapses immediately to the floor - an event commonly referred to in the industrial environment as a "demolition". It also seems to be reversible because, if the person is quickly removed to fresh air (and not injured by the fall), these individuals can sometimes be resuscitated and live again without neurological problems. Here is an agent that is not only common on the planet; Of course, it is still produced in the individual's own cells, but it is also a potent reversible inhibitor of oxidative phosphorylation that does not affect the oxygen supply.

C. Studies in murines Induction of a hibernation type state using H? S. The homeothermic animals, by definition, maintain a temperature of the central part of the body 10 to 30 ° C above the ambient temperature. For these animals to do this, they must generate heat from the energy produced by oxidative phosphorylation. The terminal enzyme complex in oxidative phosphorylation is cytochrome c oxidase. Since hydrogen sulfide inhibits this complex (Petersen, 1977; Khan et al., 1990), the inventor predicts that the exposure of a homeothermic animal to hydrogen sulphide will prevent said animal from maintaining its temperature in the central part of the body. well above room temperature. To test this hypothesis, the inventor wanted to continuously monitor the temperature of the central part of the body and the -levels of activity of a homeothermic animal (a mouse). Telemetry devices, implanted in the peritoneum of mice, can do both, and have the advantage of not introducing propensity towards readings due to the handling of mice (Briese, 1998). In addition, they can remotely monitor mice during exposure to gaseous hydrogen sulfide. Sé has previously shown that a dose of 80 parts per million (ppm) of hydrogen sulfide, is safe for mice at exposures that last up to ten weeks (CIIT 1983; Hays, 1972). Therefore, for these experiments, the inventor used a dose of 80 ppm of hydrogen sulfide to test the hypothesis. The creation of an atmosphere containing 80 ppm of hydrogen sulfide is not trivial. Over time, in the presence of oxygen, the hydrogen sulfide will be oxidized to sulfate. For that reason, for the inventor to continuously expose a mouse to an atmosphere containing 80 ppm of hydrogen sulfide, the inventor constantly mixes ambient air with a 500 ppm tank of hydrogen sulfide balanced with nitrogen. Characterization of the control of the temperature of the central part of the body. Exposure of a mouse to 80 ppm of H2S decreased its temperature from the central part to approximately two degrees Celsius above room temperature (Figure 5A). This effect was highly reproducible, since the temperature of the central part of the average body of seven mice exposed to 80 ppm of hydrogen sulfide for 6 hours followed a similar pattern (Figure 5A). The temperature of the middle part of the lowest average body of these seven mice was 15 ° C at an ambient temperature of 13 ° C. All these mice recovered successfully after reheating, when the atmosphere was changed to one containing only ambient air. As a control, the inventor replaced nitrogen with hydrogen sulfide, and did not see a substantial decrease in the temperature of the central part of the body.

Although these mice appeared to be superficially normal despite the temporary decrease in core body temperature and respiratory rate, the inventor conducted a series of behavioral tests to exclude the possibility of neurological damage from exposure to the gaseous hydrogen sulfide, the extreme reduction in the temperature of the central part of the body, the reduction in the respiratory rate, or the combination of these effects. All tests were carried out on mice before and after exposure to hydrogen sulfide. These behavior tests were selected from the SHIRPA protocol developed by the Mouse Models for Human Disease consortium (Rogers et al., 1997). There were no detectable functional differences in the mice after gas exposure. Based on this, the inventor concluded that entering a hibernation-type state is not harmful. Preliminary optimization of H2S dose. The above experiments describe the effect of 80 ppm of hydrogen sulfide on the temperature of the central part of the body of a mouse. To determine the concentration of hydrogen sulfide sufficient for the loss of thermoregulation, the inventor exposed mice at a scale of concentrations of hydrogen sulphide (20 ppm, 40 ppm, 60 ppm and 80 ppm) (Figure 6). While 20 ppm and 40 ppm of hydrogen sulfide were sufficient to cause a decrease in the temperature of the central part of the body of a mouse, it was lower compared to the reduction observed with 60 ppm and 80 ppm of hydrogen sulfide. From this experiment, the Inventor concluded that the loss of thermogenesis directly depends on the concentration of hydrogen sulfide administered to the mice. This preliminary study on the dosage scale and pharmacokinetics of hydrogen sulfide, emphasizes the need for a broader analysis. Preliminary definition of the lower limit of the temperature of the central part. The inventor is also interested in establishing a more complete understanding of the tolerance of the scale of values of the temperature of the central part of the body and the duration of time allowed in this state for the mice. The above experiments show that the inventor can repeatedly decrease the temperature of the central part of the body of a mouse to 13 to 15 ° C on request. In addition, the mice seem to tolerate the treatment for many hours. By using the same protocol, while reducing the ambient temperature, the inventor has successfully brought the temperature of the central part of the body of a mouse to 10.7 ° C (Figure 7). Other attempts will be made in the future to bring the temperature of the central part of the body even lower, and for longer periods. Although preliminary, these results demonstrate that there is a significant scale of core temperature values allowed by mouse biology, and that this scale can be explored through the loss of thermoregulation due to sulfur exposure. hydrogen. Modulation of H? S levels produced endogenously. It is well known that mammalian cells endogenously synthesize hydrogen sulfide (Wang 2002). Since this chemical is produced dynamically in the cell, it is crucial to understand the basal levels under different conditions, as this could dramatically affect the pharmacokinetics of exogenously administered hydrogen sulfide. To record this essential aspect of the present investigation, the inventor has begun to test the levels of hydrogen sulfide produced endogenously in the mouse. The inventor uses an attractive alkylation technique coupled with gas chromatography and mass specific detection to quantitate hydrogen sulfide (Hyspler et al., 2002). By using this method, the inventor observed the levels of hydrogen sulfide in undisturbed mice. Figure 8A shows that a significant amount of hydrogen sulfide exists within the mouse. In addition, the levels of hydrogen sulfide appear to depend on the ambient temperature of the mouse. Specifically, when the mice are in a cold environment, they have reduced levels of endogenous hydrogen sulfide, and when they are in a warm environment, they have increased levels of endogenous hydrogen sulfide. From this, the inventor concludes that mice regulate their levels of hydrogen sulfide in response to room temperature. Changes in endogenous levels affect the efficacy of H2S. Since the ambient temperature changes the endogenous levels of hydrogen sulfide in mice, the inventors hypothesized that the ambient temperature could firmly fix the changes in the temperature of the central part of the body upon exposure to exogenous hydrogen sulfide. The acclimatization of a mouse at low temperatures, approximately 12 ° C, creates a durable plateau that the inventor sees after the initial decrease in the temperature of the central part of the body (Figure 8B). Therefore, it seems that this acclimatization to the cold environment makes the mouse more resistant to the cooling of the central part of the body by the action of gaseous hydrogen sulfide. However, allowing the mouse to acclimate to a warm thermoneutral temperature before exposure to the gas eliminates this plateau. In fact, the normothermic mouse cooled much more rapidly when exposed to hydrogen sulfide than the mouse acclimated in a cold environment (Figure 8B). These data suggest that the endogenous levels of hydrogen sulfide in the mouse have a direct impact on the efficacy of exogenous hydrogen sulfide. H2S protects mice from hypoxia. Normal ambient air contains approximately 21% hydrogen. In a preliminary experiment exploring the protective effects of stasis on hypoxia in the mouse model, a mouse exposed to 80 ppm of hydrogen sulfide survived 11 minutes of 5.2% oxygen and 3 weeks later, was still responding well. The previously published work shows that 90% of these animals (C57BI) exposed in this way without hydrogen sulfide, does not survive (Zhang et al., 2004). This experiment included pre-equilibration of the mouse at 80 ppm of H S for 3 hours, then decreasing the oxygen tension in the chamber as described in previous experiments. The same flow rates were used as described above (ie, 500 cc / mL in a 0.5 liter chamber). It is well established for those skilled in the art that if a group of mice is exposed to 4% oxygen, 100% will die within 15 minutes. However, mice in which H2S is administered during periods when the oxygen tension is reduced to 4%, continue to be viable, even for extended periods (up to 1 hour) in these hypoxic conditions. Mice do not appear to be affected by these conditions after recovery, and are viable and usually sensitive when tested 24 hours later. This experiment differs from the previous one, because the mice were retained in the H2S environment at the end of the hypoxic exposure until the oxygen tensions were returned to normal levels (21% of 02). All compositions and methods described and claimed herein may be developed and executed without undue experimentation in light of the present disclosure. Although the compositions and methods of this invention have been described in terms of preferred embodiments, it will be apparent to those skilled in the art that variations can be applied to the compositions and methods, and in the steps or sequence of steps of the methods described. in the present, without departing from the concept, spirit and scope of the invention. More specifically, it will be apparent that certain agents that are chemically and physiologically related can substitute for the agents described herein, and that the same or similar results would be obtained. It is considered that such similar substitutes and modifications obvious to those skilled in the art, are within the spirit, scope and concept of the invention defined by the appended claims.

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Claims (134)

NOVELTY OF THE INVENTION CLAIMS

1. - The use of an oxygen antagonist to prepare a composition for inducing stasis of biological matter.

2. The use claimed in claim 1, wherein the composition comprises a sublethal dose of the oxygen antagonist.

3. The use claimed in claim 1, wherein the composition comprises an almost lethal dose of the oxygen antagonist.

4. The use claimed in claim 1, wherein the oxygen antagonist is a reducing agent.

5. The use claimed in claim 1, wherein the oxygen antagonist is a chalcogenide compound.

6. The use claimed in claim 5, wherein the chalcogenide compound comprises sulfur.

7. The use claimed in claim 5, wherein the chalcogenide compound comprises selenol.

8. The use claimed in claim 5, wherein the chalcogenide compound comprises tellurium.

9. The use claimed in claim 5, wherein the chalcogenide compound comprises polonium.

10. The use claimed in claim 4, wherein the reducing agent has a chemical structure of: where X is N, O, Po, S, Se or Te; where Y is N or O; wherein Ri is H, C, lower alkyl, a lower alcohol, or CN; wherein R2 is H, C, lower alkyl, or lower alcohol, or CN; where n is 0 or 1; where m is 0 or 1; wherein k is 0, 1, 2, 3 or 4; and wherein p is 1 or 2.

11. The use claimed in claim 10, wherein the reducing agent is a chalcogenide compound.

12. The use claimed in claim 10, wherein k is 0.

13. The use claimed in claim 10, wherein the reducing agent is selected from the group consisting of H2S, H2Se, H2Te and H2Po.

14. The use claimed in claim 10, wherein X is S.

15. The use claimed in claim 10, wherein X is Se.

16. The use claimed in claim 10, wherein X is Te.

17. The use claimed in claim 10, wherein X is Po.

18. The use claimed in claim 10, wherein X is O.

19. The use claimed in claim 14, wherein k is 0 or 1.

20. The use claimed in claim 19, wherein k is

0.

21. The use claimed in claim 10, wherein the reducing agent is DMSO, DMS, carbon monoxide, methylmercaptan (CH3SH), mercaptoethanol, thiocyanate, hydrogen cyanide, MeSH or CS.

22. The use claimed in claim 1, wherein the oxygen antagonist is a gas, semi-solid liquid or liquid.

23. The use claimed in claim 22, wherein the oxygen antagonist is a gas.

24. The use claimed in claim 23, wherein the composition is administrable through gas inhalation.

25. The use claimed in claim 23, wherein the gas comprises carbon monoxide, sulfur, selenium, tellurium or polonium, or a mixture thereof.

26. The use claimed in claim 25, wherein the gas is a chalcogenide compound.

27. The use claimed in claim 22, wherein the inhibitor is a semi-solid or liquid liquid.

28. - The use claimed in claim 27, wherein the semi-solid liquid or the liquid composition is injectable or ingested by the body.

29. The use claimed in claim 25, wherein the biological material is exposed to an amount of the composition that reduces the rate or amount of carbon dioxide production by the biological material or organism by at least about two. times.

30. The use claimed in claim 25, wherein the biological material is exposed to an amount of the composition that reduces the rate or amount of oxygen consumption by at least about twice.

31. The use claimed in claim 25, wherein the biological material is an organism, and the organism is exposed to an amount of the composition that decreases movement or mobility by at least about 10%.

32. The use claimed in claim 1, which also comprises subjecting the biological material and / or organism to a temperature-controlled environment.

33. The use claimed in claim 32, wherein the controlled temperature environment is at a non-physiological temperature - for the tissue.

34. The use claimed in claim 32, wherein the controlled temperature environment is between about -210 ° C and about 50 ° C.

35.- The use claimed in claim 34, wherein the controlled temperature environment is between approximately -210 ° C and approximately -20 ° C.

36.- The use claimed in claim 34, wherein the controlled temperature environment is between about -20 ° C and about 4 ° C.

37.- The use claimed in claim 34, wherein the controlled temperature environment is between about 0 ° C and about 50 ° C.

38.- The use claimed in claim 37, wherein the fabric reaches a temperature of the central part between 4 ° C and about 28 ° C.

39.- The use claimed in claim 37, wherein the controlled temperature environment is between about 0 ° C and about 20 ° C.

40.- The use claimed in claim 37, wherein the controlled temperature environment is between about 25 ° C and about 40 ° C.

41. The use claimed in claim 37, wherein the controlled temperature environment is between about 39 ° C and about 50 ° C.

42. The use claimed in claim 41, wherein the fabric reaches a temperature of the central part between 43 ° C and about 50 ° C.

43. The use claimed in claim 32, wherein the tissue is subjected to a controlled temperature environment before, during or after exposure to the oxygen antagonist.

44. The use claimed in claim 36, wherein the biological material is subjected to a controlled temperature environment for a period between about one minute and about one year.

45.- The use claimed in claim 32, which also comprises modulating the levels of environmental oxygen or removing the biological material or organism from an oxygen-containing environment.

46.- The use claimed in claim 1, which also comprises evaluating the level of the oxygen antagonist and / or oxidative phosphorylation in the biological material or organism.

47.- The use claimed in claim 1, which also comprises removing the oxygen antagonist.

48. The use claimed in claim 31, which also comprises increasing the ambient temperature with respect to the reduced temperature.

49. The use claimed in claim 32, wherein the oxygen antagonist is a chalcogenide compound.

50.- The use claimed in claim 25, wherein the gas is a gas mixture comprising more than one gas.

51. - The use claimed in claim 50, wherein the other gases are a non-toxic gas.

52. The use claimed in claim 40, wherein the other gases are a non-reactive gas.

53. The use claimed in claim 52, wherein the other gases are non-toxic and non-reactive.

54. The use claimed in claim 53, wherein the non-toxic and non-reactive gas is hydrogen, helium, nitrogen, neon, argon, xenon, krypton, or undoctio.

55.- The use claimed in claim 25, wherein the gas is mixed with oxygen to form a mixture of oxygen gas.

56. The use claimed in claim 55, wherein the amount of oxygen in the oxygen gas mixture is less than the total amount of the other gases in the mixture.

57. The use claimed in claim 55, wherein the gas is carbon monoxide, and the amount of carbon monoxide is almost equal to or exceeds any amount of oxygen in the oxygen gas mixture.

58.- The use claimed in claim 22, wherein the biological material or organism is exposed to the oxygen antagonist in a closed environment.

59. The use claimed in claim 58, wherein the environment oscillates at least once to a different amount of the oxygen antagonist, wherein the difference in amount is by at least one difference in percentage.

60. The use claimed in claim 59, wherein the different amount is between about 0 and 99.9% of the amount of the oxygen antagonist to which the biological material was exposed.

61.- The use claimed in claim 58, wherein the exposure of the biological material to the oxygen antagonist comprises covering or enclosing the biological material or organism with a container containing the environment, or within it.

62.- The use claimed in claim 58, which also comprises placing the biological material or organism under a vacuum.

63.- The use claimed in claim 23, wherein the biological material or organism is exposed to a normoxic environment after being exposed to the gaseous oxygen antagonist.

64.- The use claimed in claim 1, wherein the biological material is exposed to the oxygen antagonist in an environment that is at room temperature.

65.- The use claimed in claim 57, wherein the ratio of carbon monoxide to oxygen is at least about 199: 1.

66.- The use claimed in claim 65, wherein the ratio of carbon monoxide to oxygen is at least about 399: 1.

67.- The use claimed in claim 1, wherein the oxygen antagonist is administrable to the biological material or organism two or more times.

68.- The use claimed in claim 1, wherein the composition is administrable by perfusion or incubation.

69.- The use claimed in claim 1, wherein the composition is administrable by injection.

70.- The use claimed in claim 1, wherein the composition is administrable by ingestion.

71. The use claimed in claim 61, wherein the biological material or organism is perfused or incubated with the oxygen antagonist for a period of about 1 minute to about a week.

72. The use claimed in claim 71, wherein the biological material or organism is perfused or incubated with the oxygen antagonist for a period of about 5 minutes to about 24 hours.

73. The use claimed in claim 72, wherein the biological material or organism is perfused or incubated with the oxygen antagonist for a period of about 10 minutes to about 12 hours.

The use claimed in claim 73, wherein the biological material or organism is perfused or incubated with the oxygen antagonist for a period of about 30 minutes to about 6 hours.

75.- The use claimed in claim 71, where the biological matter is perfused or incubated with the oxygen antagonist for at least 2 hours.

76.- The use claimed in claim 75, wherein the biological material is perfused or incubated with the oxygen antagonist for at least 24 hours.

77.- The use claimed in claim 1, wherein the oxygen antagonist is administrable to the biological material or organism.

78. The use claimed in claim 77, wherein the oxygen antagonist is administrable to the biological material or organism intravenously, intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally intratracheally, intrarectally, intrarectally, topically, intratumorally, intramuscularly, intraocularly, subcutaneously, subconjunctivally, intravesicularly, mucosally, intrapericardially, intraumbilically, intraocularly, orally, topically, locally, by inhalation, by injection, by infusion, by continuous infusion, by localized perfusion, by means of a catheter, or by means of a lavage.

79.- The use claimed in claim 1, wherein the organism is a mammal.

80. - The use claimed in claim 79, wherein the biological material is a human.

81.- The use claimed in claim 79, wherein the mammal is a dog, a cat, a monkey, a pig, a cow, a horse, a rabbit, a rat, a mouse, a baboon or a sheep .

82.- The use claimed in claim 79, wherein the mammal has been subjected to physical trauma.

83. The use claimed in claim 1, wherein the trauma is surgery, stroke, heart attack, bone fracture, soft tissue damage, internal hemorrhage, organ damage, amputation, concussion and / or burns.

84. The use claimed in claim 1, wherein the mammal is at risk of hemorrhagic shock, or is suffering from it.

85.- The use claimed in claim 82, wherein the trauma is caused by a firearm, a fragmented grenade wound, or a knife wound.

86.- The use claimed in claim 1, wherein the biological material or organism is ill or has a disease.

87.- The use claimed in claim 86, wherein the disease is an infectious disease.

88.- The use claimed in claim 86, wherein the disease is a hyperproliferative disease.

89.- The use claimed in claim 88, wherein the disease is cancer.

90.- The use claimed in claim 86, wherein the disease is a neurodegenerative disease.

91.- The use claimed in claim 90, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease and Parkinson's disease.

92. The use claimed in claim 86, wherein the disease is an inflammatory disease.

93. The use claimed in claim 92, wherein the inflammatory disease is ulcerative colitis.

94. The use claimed in claim 92, wherein the inflammatory disease is rejection of transplantation or an autoimmune disease.

95.- The use claimed in claim 79, wherein the mammal will undergo surgery.

96.- The use claimed in claim 1, wherein the biological material is an organ, tissue or cell of the heart, lung, kidney, liver, bone marrow, pancreas, skin, bone, vein, artery, cornea, blood , small intestine, large intestine, brain, spinal cord, smooth muscle, skeletal muscle, ovary, testicle, uterus or umbilical cord.

97.- The use claimed in claim 1, wherein the biological material comprises the following types of cells: platelet, myelocyte, erythrocyte, lymphocyte, adipocyte, fibroblast, epithelial cell, endothelial cell, smooth muscle cell, cell skeletal muscle, endocrine cell, glial cell, neuron, secretory cell, cell with barrier function, contractile cell, absorption cell, mucosal cell, limb cell (cornea), mother cell, fertilized or unfertilized oocyte, or sperm .

98.- The use of a compound that has a structure of: where X is N, O, Po, S, Se or Te; where Y is N or O; wherein R ^ is H, C, lower alkyl, a lower alcohol, or CN; wherein R2 is H, C, lower alkyl, or a lower alcohol, or CN; where n is 0 or 1; where m is 0 or 1; wherein p is 1 or 2 and wherein k is 0, 1, 2, 3 or 4; to prepare a medicament for the induction of stasis in biological matter in vivo or an organism.

99.- The use claimed in claim 98, wherein the compound is a chalcogenide compound.

100.- The use claimed in claim 100, wherein the chalcogenide compound comprises sulfur.

101. The use claimed in claim 100, wherein the chalcogenide compound comprises selenium.

102. The use claimed in claim 100, wherein the chalcogenide compound comprises tellurium.

103. The use claimed in claim 100, wherein the chalcogenide compound comprises polonyl.

104.- The use claimed in claim 98, wherein k is O.

105.- The use claimed in claim 104, wherein the compound is selected from the group consisting of H2S, H2Se, H2Te and H2Po.

106.- The use claimed in claim 98, wherein X is S.

107.- The use claimed in claim 106, wherein k is 0 or 1.

108.- The use claimed in the claim 107, wherein k is O.

109. The use claimed in claim 98, wherein the compound is DMSO, DMS, carbon monoxide, methylmercaptan (CH3SH), mercaptoethanol, thiocyanate, hydrogen cyanide, MeSH or CS2. .

110.- The use of an oxygen antagonist to prepare a composition for the induction of stasis in a biological material in vivo or in an organism, wherein said composition is administrable for an effective amount of time to create hypoxic conditions for the matter biological or organism enter stasis.

111. The use claimed in claim 110, which also comprises removing oxygen from a closed environment containing the biological material or organism.

112. The use claimed in claim 111, wherein all the oxygen or part of it is replaced with another gas.

113. The use claimed in claim 112, wherein the oxygen is replaced with a gaseous oxygen antagonist.

114. The use claimed in claim 113, wherein the other gas is non-reactive and / or non-toxic.

115. The use claimed in claim 114, wherein the gas is hydrogen, helium, nitrogen, argon, neon, krypton, xenon, radon or unndoctio.

116.- The use claimed in claim 98, wherein it comprises reducing the temperature of the biological material.

117.- The use of an oxygen antagonist to prepare a composition to reduce the oxygen demand in biological matter in vivo or organism.

118.- The use of an oxygen antagonist to prepare a composition to retard the effects of a trauma on an organism.

119.- The use of an oxygen antagonist to prepare a composition for treating or preventing hemorrhagic shock in a patient.

120.- The use of an oxygen antagonist to prepare a composition to reduce the heart rate in an organism.

121. The use of an oxygen antagonist to prepare a composition for inducing hibernation in a mammal.

122. - The use of an oxygen antagonist to prepare a composition for protecting a mammal from radiotherapy or chemotherapy, wherein said composition is administrable before or during radiotherapy or chemotherapy.

123. The use of an oxygen antagonist to prepare a composition for treating a hyperproliferative disease in a mammal, wherein hyperthermia therapy is also administrable.

124. The use of an oxygen antagonist to prepare a composition for inhibiting the rejection of an organ transplant in a mammal.

125.- The use of an oxygen antagonist to prepare a composition for treating a subject with hypothermia.

126.- The use of an oxygen antagonist to prepare a composition for inducing cardioplegia in a patient suffering bypass surgery.

127.- The use of an oxygen antagonist to prepare a composition for treating a subject with hyperthermia.

128.- The use of an oxygen antagonist to prepare a composition to prevent hematological shock in a patient.

129.- The use of an oxygen antagonist to prepare a composition to promote wound healing in an organism.

130.- The use of an oxygen antagonist to prepare a composition for preventing or treating neurodegeneration in a mammal.

131. - A use for preserving an organism, which comprises administering to the organism an effective amount of an oxygen antagonist.

132.- The use claimed in claim 131, wherein the organism is preserved for future consumption.

133. The use claimed in claim 132, wherein the consumption is human consumption.

134. The use claimed in claim 133, wherein the organism is a type of shellfish. 135.- The use claimed in claim 131, wherein the organism will be used for research purposes. 136.- The use claimed in claim 135, wherein the organism is a mouse.