Correction of metabolic disorders of heart tissues and vascular walls during experimental hypertension of various geneses
Usage of various intermediates of tricarboxylic acid cycle in experimental pharmacotherapy and medical practice concerns their important role in metabolism, because they can be as energy source and substrates for synthesis; they can change direction of metabolic flows within the cell and at the extent of organism.
The basis of usage of Amber Acid and their salts includes:
· succinite-KoA deficit;
· increase of permeability of cellular and mitochondrial membranes for intermediates of tricarboxylic acid cycle;
· increase of resistance of oxidative systems of mitochondrion “charged” with substrates of tricarboxylic acid cycle towards alteration of hypoxia.
Oral injection of sodium succinate for normotensive rabbits with the dose of 100mg/kg during the period of 7 days leads to significant accumulation of pyruvate within tissues of blood - vessels under absence of significant changes of lactate content within walls of arteria and vena cava. Appropriate increase of pyruvate content gets ahead of lactate content. More feasible explanation of increase of concentration of pyruvate under injection of sodium succinate enables formation of phosphoenole pyruvate within quick cluster tricarboxylic acid cycle. Increase of pyruvate content is accompanied with reduction of relation lactate/pyruvate within cytoplasm of myocytes. Under the influence of sodium succinate the amount of lactate within tissue of heart of rats is likely to drop. Lactate content within the wall of aorta was not accompanied by changes of pyruvate level. The same as with rabbits’ intensity of oxidative metabolism within tissues of rats was subject to changes under the influence of the preparation. Significant changes of balance of metabolites under the influence of sodium succinate may be conditioned by differences of formation of phosphoenole pyruvate within tissues of rabbits and rats.
Monitoring of changes of balance of catecholamines for rats under the influence of sodium succinate were as follows: accumulation of adrenaline in the wall of major arterial blood vessels for rabbits, accumulation of noradrenaline by the tissues of carotid artery and aorta of thorax; accumulation of adrenaline by the heart and renal. The present facts may be conditioned by the syntheses of catecholamines within chromaffin tissue due to direct energizing impact of sodium succinate towards cellular mitochondrion. Increase of content of catecholamines is an important link of mediated metabolic influence of sodium succinate. Sodium succinate evokes significant accumulation of pyruvate within tissues of blood vessels of rabbits with pyruvate hypertension, glucose within wall of aorta, reduction of lactate level within all except femoral artery and vascular segments. Shift of cytoplasmic relation lactate/pyruvate has been observed.
Increase of pyruvate content is typical test of impact of sodium succinate under metabolic acidosis. Similar impact of the preparation although expressed in other level is being observed for rats with pituitrin hypertension who has increased level of pyruvate in heart tissues due to impact of sodium succinate. Rats contrary to rabbits had had significantly increased lactate level within heart tissues and aorta. The preparation is likely to normalize redox system of pyridine nucleotides within cardial and kvazi - myocytes. Adenyl energy charge within cells of heart is likely to restore till control level.
Sodium succinate for rats with pituitrin hypertension acts the more favourably towards cardial metabolism compared with aorta. If level of adrenaline normalizes, wall of aorta will contain increase of catecholamine content on the basis of increase of adrenaline and noradrenaline within the tissues of renal gland. The preparation has no “pyruvatogenic” effect on the basis of initial accumulation of pyruvate by vascular tissue of rabbits. Despite of aerobiotic shift within cytoplasm of myocytes of rabbits with reno-vascular hypertension, significant drop of inorganic phosphate and pool of adenyl within vascular wall stops consideration about high energy state of the system.
Increase of pyruvate content for rats with reno-vascular hypertension under their treatment with sodium succinate has been observed in line with drop of lactate level within vascular wall. Under the treatment of rats with salt hypertension level of pyruvate dropped within aorta in line with increase of the amount of lactate within the heart and blood. Balance of noradrenaline within wall of aorta disturbed as a result of pathology was not likely to restore after intake of sodium succinate. Increase of adrenaline and noradrenaline content within renal grand is not the result of accumulation of adrenaline within the heart and aorta. But level of noradrenaline within the heart is likely to increase. In the event of stress-inducing hypertension for rats sodium succinate influenced reduction of pyruvate and lactate content within heart and aorta tissues. For rats with pituitrin hypertension sodium succinate was likely to restore level of intracellular and total Na on the basis of preservation of low level of K at the extent of cells. By normalizing Na level at the extent of cell and plasma, the preparation strengthens cellular fixation of Na for rats with salt hypertension. Combination of papaverine and sodium succinate for rabbits with pituitrin hypertension (1 mg/kg of weight 7 days subcutaneously) significantly changed character of metabolic processes within vascular tissue of animals with hypertension. Level of pyruvate and lactate was likely to increase within vascular wall under normalization of content of inorganic phosphate and aerobic shift of redox-system. Impact of sodium succinate occurs under its combination with papaverine. Evaluation of metabolic effectiveness of sodium succinate has confirmed capability of new preparation to eliminate metabolic acidosis, occurring within vascular system under experimental hypertension of various geneses. Achieved results on increase of pyruvate content within tissues of animals after injection of sodium succinate should be used for involving of the present preparation into complex treatment under conditions of hypertension.
Authors
Ratchkov A.K. Ratchkova M.A
Correction of energy balance as condition of optimization of immuno-metabolic processes for newborn during inflammatory occurences
Infant mortality, in many cases, is related with lethal complications for newborns, where mortality because of sepsis or pneumonia has an important role. Cells protective mechanism of newborns’ organism is insufficiently able to protect from microbes and their toxins, moreover lymphatic system is immature. That is the main reason of difficult treatment of infectious inflammatory process. Protein synthesis function and reactions of energy balance are educed among metabolic processes, which determine resistance for negative factors. We investigated remedial effect of preparations, which have influence on energy balance, particularly Amber Acid, natrium, levamizole and splenin.
Liquor of 15% sodium succinate was prescribed for newborns, which have pneumonia, according to their weight in 150mg/kg. Remedial effect of preparation is characterized as more rapid intoxication symptoms, release of respiration insufficiency, hypoxia and disorder of microcirculation. Releases of hypodinamic processes perioral cyanosis, grey color of skin and its marble undertone and reduction of heart tones are increased. Catarrhal symptoms of upper airway are eliminated sooner; admission period of patients becomes shorter. In the case of complicated pneumonia (20 newborns) hospitalization period became shorter from 25±0,4 to 23,5±0,43 (р<0,01),>21±0,17 days, р<0,05) and toxicosis (10,51±0,21 and 12,74±0,14 days, р<0,02).
Comparison of results, achieved under identification of indicators of energy balance have showed significant increase of activity of mitochondrion enzyme of succinate dehydrogenase within lymphocytes (20, 18±0, б2 versus 17, 22±0, 54 of formazan granules on a single lymphocyte, р<0,005) and also slightly increased ATP level (492, 17±17, 21 versus 48б, 25±26, 34 mkmol/g, р<0,05). The present facts evidence activation of oxidation processes, accompanying improvement of energy production under the influence of sodium succinate.
Analysis of acid-base balance, carried out one week later since the beginning of the therapy have showed less expressed disorders in the basic group compared with the control group. Before discharge from the hospital, stable normalization of acid-base balance has been observed with 90% of children, treated with sodium succinate which significantly gets ahead of indicators of control group (77%). More well-marked influence of sodium succinate towards dynamics of the process and indicators of energy balance has been observed under its application in the event of prolonged course of pneumonia treatment. Appetite and well-being have been significantly improved already on the 2nd week in line with activation of motion behavior. Physical changes used to become less long-lasted (15, 31±0, 13 versus 20, 51±0, 29, р<0, 02). Restoration of normal cardial activity occurred significantly with children from the basic group (15, 44±0, 31) compared with control group (17, 16±0, 32), р<0, 05. Average weight increase during the period of treatment for children treated with sodium succinate comprised б84±50g versus children of control group 320±40g, (р<0, 05).
Achieved results enable to assume about significant therapeutical effectiveness of sodium succinate and give ground to recommend it for clinical application during the treatment of pneumonia for children.
Authors
Reznik B.I Krainaja Z.V
Usage of sodium succinate for treatment of acute respiratory –viral infections for children
Shortage of effective etiotropic antiviral remedies causes necessity to search for preparations, stimulating protection of human body against infections and capable of facilitating course of acute respiratory viral infections for children in line with reducing risk of complications. Experimental researches have proved favourable impact of sodium succinate under different pathologic conditions, being realized probably due to energy processes within mitochondrion. Successful intake of sodium succinate for children of tender age with bronchial allergy and pneumonia provided basis for usage of the latter preparation under uncomplicated and complicated variations of acute respiratory – viral infections (ARVI) for children. Advantages of sodium succinate are as follows: harmlessness of the preparation, absence of cumulative features and good tolerance. The preparation is being administrated for children in compliance with licence of Pharmaceutical Committee (minute No 3 of 30 July of 1972 USSR).
141 children at the age from 2 to 5 years suffering from uncomplicated and complicated cases of SRVI have been passes under doctors’ monitoring at the clinics of pediatric infections (Leningrad). Sodium succinate has been involved in usual pathogenic therapy for 108 children during the course of acute period of disease; 33 of patients have not been treated with sodium succinate and comprised the comparative group. Laboratory diagnostics of ARVI for children has been performed involving generally accepted serological and immunofluorescence methods with additional definition of classes of specific immunoglobulins M and G in a way of destruction of macroglobulins using preparation of mercamine group. Using the method of quantitative hemadsorption within original modification and heterospecific thermolabile beta – inhibitors in Merskey’s reaction with virus of Newcastle disease, interferon has been determined of all the children. Determination of contents of secretory immunoglobulin has been carried out according to Manchini. Percentage and absolute number of T-lymphocytes within one millilitre of blood have been determined for 141 children during dynamics of ARVI according to the reaction of spontaneous rosetting with erythrocytes of sheep and B - cells using rosetting reactions with zymosan and complement. Percentage of T and B rosettes in the preparation has been counted under absolute quantity. Therapeutical effectiveness of sodium succinate has been estimated according to the following characteristics of ARVI: height and duration of temperature, duration of intoxication and catarrhal syndrome and frequency of development of bacterial complications. History of pediatric diseases with defined etiology of disease has been involved into analysis.
Maximal body temperature of patients treated with sodium succinate, duration of temperature reaction and intoxication was the same as indicators of comparative group. Substantial differences between the groups have been observed only at the extent of catarrhal syndrome. For the group of children suffering from uncomplicated influenzal infection being treated with sodium succinate rhinitis was likely to reduce on average by 2.8±0.6 days and for the group without intake of the preparation by 5.1±0.8 days. Under uncomplicated single agent infections of various genesis throat hyperemia involving treatment sodium succinate continued for б,2±0,52 days in comparative group – for 8,5±0,4 (р<0,05) days. In the event of uncomplicated combined ARVI throat hyperemia accordingly continued for 5,3±1,67 days and for 10,2±0,86 days (р<0,01). Differences of frequency of delayed bacterial complications also have been observed: disease of patients treated with sodium succinate resulted in complication of pneumonia and bronchitis 3 times less as comparative group (5,2% versus 16,6%). Substantial reduction of superinfection under conditions of in-patient hospital has also been observed. Reduction of continuation of catarrhal syndrome, reduction of frequency of bacterial complications can be explained as stimulation of local immune system mostly production of secretory immunoglobulin A (Ig A).
Results, achieved in defining IgA for children by using method of reliable intervals, have been subdivided into thee groups: low IgA concentration (5–30 mg/l) – 52 children, medium - (31–80 mg/l) – 10 children and high - (81 mg/l) 20 children. The group of children being treated with usual pathogenic therapy (24 children in the group) had 17 children with low IgA concentration, 3 with medium IgA concentration and 4 with high concentration. Low IgA concentration during the course of disease in the comparative group remained almost the same: 11,7±0,2 at the beginning and 10,9±0,4 mg/l during recovery. In the event of medium and high primary IgA concentration significant drop has been observed during the period of recovery, which 4-7 times gets ahead of results from comparative group. In the event of high IgA concentration children treated with sodium succinate were likely to have lower amount of IgA concentration in their nasal phlegm but nevertheless such reduction has been less expressed as for children of comparative group (64,4±6,0 mg/l and 45,5±6,1 mg/l, р<0,05).
Under impact of sodium succinate the amount of T – lymphocytes significantly increased not only in the group of uncomplicated ARVI, but also in development of complications (from 889±39 to 1217±57, р<0,05). Significant increase of the amount of B – cells has been observed only in the event of complicated ARVI for children treated with sodium succinate (from 285±11 to 337±19, p<0,5). Indicators of the state of non-specific resistance for majority of patients treated with sodium succinate have almost remained the same during the course of disease. The amount of thermolabile beta – inhibitors during the period of recovery has not increased their content during the period of acute respiratory viral infection.
Increase of the level of serumal interferon has been observed during dynamics of ARVI for children treated with sodium succinate from 5,2 to 10 units/ml (р<0,05).
But as it is known concentration of interferon less than 10 units/ml is not enough for relief of clinical evidence of ARVI. Thus the present researches have proved out advisability of intake of sodium succinate as the component of complex treatment for children, suffering from ARVI.
Authors: Ivanova V.V, Kurbatova G.P, Blagoslovenskij G.S, Aksenova O.A.
Acid – natural preparation for diabetes treatment
In the case of diabetes dependent on insulin, hyperglycemia goes together with other metabolic disorders: hyperlipidemy, hyperlactatemia, and ketosis. Decrease of carbohydrate on purpose to egest hyperglycemia is the main principle of diabetes treatment. The initial criterion of this compensation is absence of sugar in the urine and reduced glucose concentration in the blood during twenty four hours. Such correction of diabetes is carried out by choosing diet, injecting insulin, taking non-hormonic preparations, which reduces amount of sugar or its combinations in the blood.
The usage of glucose in peripheral tissues changes dependent on the concentration of free fatty acid and ketotic corpuscules in blood plasma. Any increase of fatty acid or ketotic corpuscules is a reason of suppression of glucose usage in the tissues. It is visible in the case of both normal and enlarged concentration of insulin. By contrast, injection of glucose reduces oxidation speed of fatty acid and enlarges usage of glucose in peripheries. That is to say, that reduction of fatty acid and ketonic corpuscules level in blood plasma may stimulate usage of glucose in the tissues and reduce amount of glucose in plasma.
In the course of our experiments we noticed that progress of diabetes growth for rats (after injections of alloxan) depends on process of lipid circulation a lot according to glucose concentration in the blood. It was proved that animals, which had injections of alloxan, could be divided into two groups in unison called “diabetic” and “non-diabetic”. Sure enough, injections of alloxan caused increase of glucose in the blood till the trespass of norm for some rats, for others concentration of glucose remained almost unchanged.
Table 1.
Parallel changes of glucose concentration and ketotic corpuscules in the blood of rats causing diabetes with alloxan
Dimensional rate
Groups
Control (n–18)
After injection of alloxan
diabetic (n–10)
non-diabetic (n–10)
Glucose
9,8±0,4
18,0±1,5
8,7±0,7
3-hidroxibutirate
127±6
433±66
127±14
Acetoacetate
89±6
222±32
93±10
Amount of ketonic corpuscules
213±11
651±91
212±7
Note. 40 mg/kg of alloxan was injected in proportion to 1 ml saline. Saline was injected for the control group. Three days later, concentration of indicated substances in the blood was measured. Glucose, 3-D –hidroxibutirate and acetoacetate was measured by fluorimetric fermentatic methods. For each alloxan group rats were selected in tens. Criterion for the selection was glucose concentration in the blood: under 10 mg and above 15 mg for “diabetic” and “non-diabetic” groups.
It is shown in the table that level of glucose concentration in the blood does not rise (diabetes does not develop0 when injection of alloxan does not cause increase of ketotic corpuscules in the blood. That is to say, such animals do not differ from controlled, their weight does not decrease and there is no polyury. Those animals may be ascribable to “non-diabetic” group. In other group of rats, “diabetic”, increase of glucose concentration and ketotic blood corpuscules, which are caused by alloxan, may be seen. A weight of these rats was 20% lower than a weight of control group. The former drank a lot of water and the polyury was noticed. Thus, the positive correlation between glucose and amount of ketotic corpuscules in the blood was a feature of rats of “diabetic” group. Spanish researches, who are working with Spreg-Doyli rats causing diabetes with alloxan, got similar (although not identical) results. Glucose accession into peripheral tissues, for example into skeletal muscles and erythrocytes, is a process dependent on insulin, whereas ketotic corpuscules spared over the tissues freely and are independent of insulin. Thus, the findings let to guess, that having diabetes, when permeability of cell membrane disconcerts because of lack of insulin, the amount of glucose concentration in the blood may be diminished by the reduction of concentration of ketotic corpuscules.
We have investigated an impact of various natural preparations on concentration of ketotic corpuscules in the blood, and identified that sodium succinate the most effectively reduces concentration of ketotic corpuscules. It, especially, increases when the animal is starving (Table 2). After twenty four hours of starving concentration of 3-D –hidroxibutirate in blood plasma of the rats used to rise to 1,95 mg, acetoacetate – to 0,5mg (norm approximately 0,2 and 0,05mg respectively), how ever the effect of sodium succinate used to reduce concentration of ketotic corpuscules significantly (to 1,0 and 0,38mg respectively) during the thirty minutes previous to animal death.
Table 2.
An impact of sodium succinate on the concentration of ketotic corpuscules in the blood of starving rats
Dimensional metabolites
Concentration, mg
Saline
Sodium succinate
3-O-hidroxibutirate
1,95±0,25
1,00±0,10*
Acetoacetate
0,51±0,10
0,38±0,12
Amount of ketotic corpuscules
2,44±0,33
1,39±0,19*
Note. Rats (in tens in each group) were starving twenty-four hours. Afterwards at 9hr 30min saline or sodium succinate (50mg/kg) was injected for them. After 30 min level of concentration of ketotic corpuscules was researching by the fluorimetric fermentatic method. Deviations of average value and average effective values from normal, *р<0,005, in comparison with control group, are listed.
Mechanism, through which sodium succinate may reduce concentration of ketonic corpuscules and glucose, is indeterminate. It is may be guessed, that oxidation of succinate, which may intensify metabolism in vivo, creates surplus amount of ATP, witch is sufficient for the activity of acetoacetate, lipid biosynthesis, herewith for the stimulation of glucose transmission to peripheral tissues.
We are inclined to think, that sodium succinate may be used for reduction of glucose amount in the blood after causing diabetes with alloxan for animals. Tentative experiments proved that impact of sodium succinate (for animals diabetes was caused by alloxan) reduces not only amount of ketotic corpuscules in blood plasma, but also glucose concentration (more effectively).
Impact of sodium succinate on glucose and ketotic corpuscules concentration in rat blood (diabetes for them was caused by alloxan).
Metabolites concentration in the blood
Saline
Sodium succinate
Glucose
30±4
16,4±4,2
3-O-hidroxibutirate
886±85
530±89
Acetoacetate
369±63
298±60
Amount of ketonic corpuscules, мкМ
1250±142
824±204
Note. Alloxan was injected for the rats in 40mg/kg. Three days later, sodium succinate (50mg/kg) was injected for one group or rats, saline – for other group. After half-an-hour all animals were decapitated and metabolites, which are described in table 1) found.
Deviations of average value and average effective values from normal were listed.
In contradistinction to insulin, such unnatural agents like sulphacarbamide may lower glucose amount in plasma without its entrance into tissues stimulation and without utilization in the tissues. In that case the application of preparations, this reduces glucose concentration in the blood through reduction of ketotic corpuscules concentration, stimulation of glucose transmission to cells and its utilization in peripheral tissues without inhibition of gluconeogenesis, may be beneficial.
Authors: E. A Kosenka, J.G. Kaminskij, M.N. Kondrasheva.
The Institute of Theoretic and Experimental Biophysics
Use of Amber Acid and other components of energy balance for treatment of oncological cases
German physiologist Otto Warburg set up the hypothesis, which claims that the cause of cancer can be found in the absence of oxygen. It happens due to various reasons, and is considered to be the first step towards cellular disorders. Those disorders may become malignant. In the case of permanent insufficiency of oxygen one part of the cells perishes, another part adapts to the newly formed conditions. The latter part of the cells causes a great activity of glycolysis and like so it fills energy deficit. According to Warburg’s mind, respiration disorder is the basis of re-formation of healthy cells to cancerous. Such a disorder may emerge in consequence of various reasons. The most important of them is the disorder of oxygen-supply.
Nevertheless, not every respiration disorder causes cancer. These are the conditions that cause malignant disorders:
· Respiration disorder must be reversible, i.e. it must remain after the extraction of agent, which caused the disorder.
· Respiration disorder is not supposed to be such as to cause cells losses, according to Warburg, a sudden loss of oxygen or high concentration of toxic are more dangerous than low and permanent influence of these factors.
· Respiration disorder must remain during the cell division, i.e. it becomes genetically descendible.
· Respiration disorder cannot have influence on glycolysis.
· Respiration disorder should not rebound on cellular respiration.
In that case, respiration disorder may become carcinogenic.
The conversion of normal cells into cancerous comes along in two stages. In the first stage, after the occurence of respiration disorder, long period of the struggle for existence of cell begins. It is true to think that the only latent period of cancer formation is none other than period, during which the selection of cells happens and the glycolysis of the tissue intensifies, after the respiration disorder. Respiration disorder may be associated with the decrease number of mitochondrions in the cell.
Warburg thought that respiration of cancerous cells is not just simply weak, but weak in comparison with really intensive aerobic glycolysis. Thus, the respiration of cancerous cells becomes inferior with regard to quality (stopping of respiration, oxidation and phosphorilation). Warburg’s theory was criticized more than once. Till now the question, weather disorder of mitochondrions may be a reason of the conversion of normal cell into cancerous, is open. The main goal of this work is the testing of the state cells mitochondrions and search for the influence on cancerous cells. This, in turn, lets heal the owner of those cells, carrier of cancer.
There were exams on three types of respiration of human’s cancerous cells and ultra-structures: specifically, ovarian cancer (OV), stomach cancer (CaVe) and mesenchymoma of connective tissue (Sa19). It is proven that great activity of glucose and weak respiration of the OV cells comes to the top. A number of mitochondrions, compared together with normal cells, decreases there. According to their external look, they were similar to mitochondrions, which are affected by the unhealthy agent for the respiration – cyanide: mitochondrions were 2 – 3 times longer. CaVe and Sa19 were breathing, though glucose was increased in them also. Populations of mitochondrions were heterogeneous in Sa19 cells: a lot of swollen mitochondrions. Majority of organoids were in, known as, mediate (with the optimum amount of energy) state. Small, newly formed mitochondrions are also observed. The facts suggest that activity of mitochondrions population in Sa19 cell is high. The look of CaVe cells and their mitochondrions is similar to Sa19, but the former are not so active and pro-mitochondrions can be seen only after irradiation of the tissue.
Table 1
Rapidity of respiration, rates of conjugation of mitochondrions and amount of ATP in cells of Sa19 mitochondrions.
Substratum of oxidation
V3
V4
АDP/О
Control of respiration (by Chance)
Alpha-ketoglutarate
2,50±0,03
1,75±0,04
2,1±0,1
2,0±0,3
Amber Acid
2,00±0,05
0,76±0,06
1,8±0,2
2,6±0,1
Table 2Amount of ATP in cells of Sa19 mitochondrions
Amount of endogenous ATP (mg/mg protein)
Amount of ATP after addition of synthesized ADP (mg/mg protein)
10,1±2,1
362,6±32,4
As shown in the Table 1 and Table 2, mitochondrions, which are abstracted from Sa19 cells, has a high rapidity of respiration, but rates of conjugation are reduced in them. Amount of endogenous ATP is lesser than in mitochondrions of normal tissue. Synthesis of ATP is fairly active.
Another set of experiments shows the rates of conjugation of cells respiration and mitochondrions. They are abstracted from rat hepathoma 22, from liver of rats, which has tumour, from laboratorially affected liver of rats (the expansion of tumor stopped for them after vaccination) also from not affected liver of rats.
Table 3
Respiration of mitochondrions, which are abstracted form various liver of rats. Substratum of oxidation – glutamine acid (5 mg/ml).
Source of mitochondrions
V3
V4
АDP/О
Control of respiration (by Chance)
1.Hepathoma (n=6)
0,85±0,05
0,42±0,06
2,1±0,2
2,0±0,1
2. Liver of rats, which has tumour (n=6)
0,30±0,03
0,12±0,02
2,2±0,3
2,5±0,2
3. Laboratorially affected liver of rats (n=3)
0,74±0,02
0,20±0,04
2,8±0,04
3,7±0,4
4.Norm (n=10)
0,49±0,01
0,15±0,02
2,6±0,1
3,3±0,2
P1-2<0,05 p3-2<0,05 p3-1<0,05 p3-4<0,05
From the Table 3 it may be noticed that rapidity of respiration of mitochondrions, which are abstracted from hepathoma, is rather higher in the liver of rats, which has tumour. Though, the rates of conjugations are almost the same. We are of the opinion that it reports about the activity of energy reactions in hepathoma (conjugation of variable mitochondrions) and their reduction in mitochondrions of liver of rats, which has tumour. High rapidity of respiration and great rates of conjugation is characteristic of mitochondrions of liver of rats affected under conditions of laboratory.
The count, basically, confirms Warburg’s hypothesis, analyzing respiration disorder of cancerous cells in a wider meaning: like respiration changes and energy reactions. Therein lies, that it is possible to try to cure cancerous cells, by regeneration normal circulatory system. Herewith it helps organism, which carries cancer, to fight against this trouble.
A group of doctors of Moscow 40th hospital followed our recommendations for usage of Amber Acid (AA) together with citric acid, alpha-ketoglutarate acid, vitamins and some other medical plants. This treatment was for volunteers. During the course of treatment patients were on almost vegetarian, anti-diabetic diet. They were taking vitamins together with special medical agents of treatment. Observation period was 5 years. Patients of 2-3 stage of disease were supervised. Treatment proceeded till and after the radical operation.
Table 4
Influence of Amber Acid (together with citric acid and 2-oxo-glutaric acid on mortality of oncological patient in a course of five years (%).
Type of tumour
Conditions of experiment
Number of patients
Mortality in a course of 5 years, %
Cervical cancer
Control
Treatment
5
10
80%
10%
Uterine myoma
Control
Treatment
10
15
50%
0%
Ovarian cancer
Control
Treatment
6
8
90%
30%
Cancer of intestine and rectum
Control
Treatment
10
10
80%
10%
Mammary cancer
Control
Treatment
10
20
60%
10%
The facts suggest that AA may be successfully used for healing oncological patients other type of results of circulatory system disorder.
Author: E.V. Kozyreva
Inst. of Biophysics, Moscow, Russian Federation
Usage of various intermediates of tricarboxylic acid cycle in experimental pharmacotherapy and medical practice concerns their important role in metabolism, because they can be as energy source and substrates for synthesis; they can change direction of metabolic flows within the cell and at the extent of organism.
The basis of usage of Amber Acid and their salts includes:
· succinite-KoA deficit;
· increase of permeability of cellular and mitochondrial membranes for intermediates of tricarboxylic acid cycle;
· increase of resistance of oxidative systems of mitochondrion “charged” with substrates of tricarboxylic acid cycle towards alteration of hypoxia.
Oral injection of sodium succinate for normotensive rabbits with the dose of 100mg/kg during the period of 7 days leads to significant accumulation of pyruvate within tissues of blood - vessels under absence of significant changes of lactate content within walls of arteria and vena cava. Appropriate increase of pyruvate content gets ahead of lactate content. More feasible explanation of increase of concentration of pyruvate under injection of sodium succinate enables formation of phosphoenole pyruvate within quick cluster tricarboxylic acid cycle. Increase of pyruvate content is accompanied with reduction of relation lactate/pyruvate within cytoplasm of myocytes. Under the influence of sodium succinate the amount of lactate within tissue of heart of rats is likely to drop. Lactate content within the wall of aorta was not accompanied by changes of pyruvate level. The same as with rabbits’ intensity of oxidative metabolism within tissues of rats was subject to changes under the influence of the preparation. Significant changes of balance of metabolites under the influence of sodium succinate may be conditioned by differences of formation of phosphoenole pyruvate within tissues of rabbits and rats.
Monitoring of changes of balance of catecholamines for rats under the influence of sodium succinate were as follows: accumulation of adrenaline in the wall of major arterial blood vessels for rabbits, accumulation of noradrenaline by the tissues of carotid artery and aorta of thorax; accumulation of adrenaline by the heart and renal. The present facts may be conditioned by the syntheses of catecholamines within chromaffin tissue due to direct energizing impact of sodium succinate towards cellular mitochondrion. Increase of content of catecholamines is an important link of mediated metabolic influence of sodium succinate. Sodium succinate evokes significant accumulation of pyruvate within tissues of blood vessels of rabbits with pyruvate hypertension, glucose within wall of aorta, reduction of lactate level within all except femoral artery and vascular segments. Shift of cytoplasmic relation lactate/pyruvate has been observed.
Increase of pyruvate content is typical test of impact of sodium succinate under metabolic acidosis. Similar impact of the preparation although expressed in other level is being observed for rats with pituitrin hypertension who has increased level of pyruvate in heart tissues due to impact of sodium succinate. Rats contrary to rabbits had had significantly increased lactate level within heart tissues and aorta. The preparation is likely to normalize redox system of pyridine nucleotides within cardial and kvazi - myocytes. Adenyl energy charge within cells of heart is likely to restore till control level.
Sodium succinate for rats with pituitrin hypertension acts the more favourably towards cardial metabolism compared with aorta. If level of adrenaline normalizes, wall of aorta will contain increase of catecholamine content on the basis of increase of adrenaline and noradrenaline within the tissues of renal gland. The preparation has no “pyruvatogenic” effect on the basis of initial accumulation of pyruvate by vascular tissue of rabbits. Despite of aerobiotic shift within cytoplasm of myocytes of rabbits with reno-vascular hypertension, significant drop of inorganic phosphate and pool of adenyl within vascular wall stops consideration about high energy state of the system.
Increase of pyruvate content for rats with reno-vascular hypertension under their treatment with sodium succinate has been observed in line with drop of lactate level within vascular wall. Under the treatment of rats with salt hypertension level of pyruvate dropped within aorta in line with increase of the amount of lactate within the heart and blood. Balance of noradrenaline within wall of aorta disturbed as a result of pathology was not likely to restore after intake of sodium succinate. Increase of adrenaline and noradrenaline content within renal grand is not the result of accumulation of adrenaline within the heart and aorta. But level of noradrenaline within the heart is likely to increase. In the event of stress-inducing hypertension for rats sodium succinate influenced reduction of pyruvate and lactate content within heart and aorta tissues. For rats with pituitrin hypertension sodium succinate was likely to restore level of intracellular and total Na on the basis of preservation of low level of K at the extent of cells. By normalizing Na level at the extent of cell and plasma, the preparation strengthens cellular fixation of Na for rats with salt hypertension. Combination of papaverine and sodium succinate for rabbits with pituitrin hypertension (1 mg/kg of weight 7 days subcutaneously) significantly changed character of metabolic processes within vascular tissue of animals with hypertension. Level of pyruvate and lactate was likely to increase within vascular wall under normalization of content of inorganic phosphate and aerobic shift of redox-system. Impact of sodium succinate occurs under its combination with papaverine. Evaluation of metabolic effectiveness of sodium succinate has confirmed capability of new preparation to eliminate metabolic acidosis, occurring within vascular system under experimental hypertension of various geneses. Achieved results on increase of pyruvate content within tissues of animals after injection of sodium succinate should be used for involving of the present preparation into complex treatment under conditions of hypertension.
Authors
Ratchkov A.K. Ratchkova M.A
Correction of energy balance as condition of optimization of immuno-metabolic processes for newborn during inflammatory occurences
Infant mortality, in many cases, is related with lethal complications for newborns, where mortality because of sepsis or pneumonia has an important role. Cells protective mechanism of newborns’ organism is insufficiently able to protect from microbes and their toxins, moreover lymphatic system is immature. That is the main reason of difficult treatment of infectious inflammatory process. Protein synthesis function and reactions of energy balance are educed among metabolic processes, which determine resistance for negative factors. We investigated remedial effect of preparations, which have influence on energy balance, particularly Amber Acid, natrium, levamizole and splenin.
Liquor of 15% sodium succinate was prescribed for newborns, which have pneumonia, according to their weight in 150mg/kg. Remedial effect of preparation is characterized as more rapid intoxication symptoms, release of respiration insufficiency, hypoxia and disorder of microcirculation. Releases of hypodinamic processes perioral cyanosis, grey color of skin and its marble undertone and reduction of heart tones are increased. Catarrhal symptoms of upper airway are eliminated sooner; admission period of patients becomes shorter. In the case of complicated pneumonia (20 newborns) hospitalization period became shorter from 25±0,4 to 23,5±0,43 (р<0,01),>21±0,17 days, р<0,05) and toxicosis (10,51±0,21 and 12,74±0,14 days, р<0,02).
Comparison of results, achieved under identification of indicators of energy balance have showed significant increase of activity of mitochondrion enzyme of succinate dehydrogenase within lymphocytes (20, 18±0, б2 versus 17, 22±0, 54 of formazan granules on a single lymphocyte, р<0,005) and also slightly increased ATP level (492, 17±17, 21 versus 48б, 25±26, 34 mkmol/g, р<0,05). The present facts evidence activation of oxidation processes, accompanying improvement of energy production under the influence of sodium succinate.
Analysis of acid-base balance, carried out one week later since the beginning of the therapy have showed less expressed disorders in the basic group compared with the control group. Before discharge from the hospital, stable normalization of acid-base balance has been observed with 90% of children, treated with sodium succinate which significantly gets ahead of indicators of control group (77%). More well-marked influence of sodium succinate towards dynamics of the process and indicators of energy balance has been observed under its application in the event of prolonged course of pneumonia treatment. Appetite and well-being have been significantly improved already on the 2nd week in line with activation of motion behavior. Physical changes used to become less long-lasted (15, 31±0, 13 versus 20, 51±0, 29, р<0, 02). Restoration of normal cardial activity occurred significantly with children from the basic group (15, 44±0, 31) compared with control group (17, 16±0, 32), р<0, 05. Average weight increase during the period of treatment for children treated with sodium succinate comprised б84±50g versus children of control group 320±40g, (р<0, 05).
Achieved results enable to assume about significant therapeutical effectiveness of sodium succinate and give ground to recommend it for clinical application during the treatment of pneumonia for children.
Authors
Reznik B.I Krainaja Z.V
Usage of sodium succinate for treatment of acute respiratory –viral infections for children
Shortage of effective etiotropic antiviral remedies causes necessity to search for preparations, stimulating protection of human body against infections and capable of facilitating course of acute respiratory viral infections for children in line with reducing risk of complications. Experimental researches have proved favourable impact of sodium succinate under different pathologic conditions, being realized probably due to energy processes within mitochondrion. Successful intake of sodium succinate for children of tender age with bronchial allergy and pneumonia provided basis for usage of the latter preparation under uncomplicated and complicated variations of acute respiratory – viral infections (ARVI) for children. Advantages of sodium succinate are as follows: harmlessness of the preparation, absence of cumulative features and good tolerance. The preparation is being administrated for children in compliance with licence of Pharmaceutical Committee (minute No 3 of 30 July of 1972 USSR).
141 children at the age from 2 to 5 years suffering from uncomplicated and complicated cases of SRVI have been passes under doctors’ monitoring at the clinics of pediatric infections (Leningrad). Sodium succinate has been involved in usual pathogenic therapy for 108 children during the course of acute period of disease; 33 of patients have not been treated with sodium succinate and comprised the comparative group. Laboratory diagnostics of ARVI for children has been performed involving generally accepted serological and immunofluorescence methods with additional definition of classes of specific immunoglobulins M and G in a way of destruction of macroglobulins using preparation of mercamine group. Using the method of quantitative hemadsorption within original modification and heterospecific thermolabile beta – inhibitors in Merskey’s reaction with virus of Newcastle disease, interferon has been determined of all the children. Determination of contents of secretory immunoglobulin has been carried out according to Manchini. Percentage and absolute number of T-lymphocytes within one millilitre of blood have been determined for 141 children during dynamics of ARVI according to the reaction of spontaneous rosetting with erythrocytes of sheep and B - cells using rosetting reactions with zymosan and complement. Percentage of T and B rosettes in the preparation has been counted under absolute quantity. Therapeutical effectiveness of sodium succinate has been estimated according to the following characteristics of ARVI: height and duration of temperature, duration of intoxication and catarrhal syndrome and frequency of development of bacterial complications. History of pediatric diseases with defined etiology of disease has been involved into analysis.
Maximal body temperature of patients treated with sodium succinate, duration of temperature reaction and intoxication was the same as indicators of comparative group. Substantial differences between the groups have been observed only at the extent of catarrhal syndrome. For the group of children suffering from uncomplicated influenzal infection being treated with sodium succinate rhinitis was likely to reduce on average by 2.8±0.6 days and for the group without intake of the preparation by 5.1±0.8 days. Under uncomplicated single agent infections of various genesis throat hyperemia involving treatment sodium succinate continued for б,2±0,52 days in comparative group – for 8,5±0,4 (р<0,05) days. In the event of uncomplicated combined ARVI throat hyperemia accordingly continued for 5,3±1,67 days and for 10,2±0,86 days (р<0,01). Differences of frequency of delayed bacterial complications also have been observed: disease of patients treated with sodium succinate resulted in complication of pneumonia and bronchitis 3 times less as comparative group (5,2% versus 16,6%). Substantial reduction of superinfection under conditions of in-patient hospital has also been observed. Reduction of continuation of catarrhal syndrome, reduction of frequency of bacterial complications can be explained as stimulation of local immune system mostly production of secretory immunoglobulin A (Ig A).
Results, achieved in defining IgA for children by using method of reliable intervals, have been subdivided into thee groups: low IgA concentration (5–30 mg/l) – 52 children, medium - (31–80 mg/l) – 10 children and high - (81 mg/l) 20 children. The group of children being treated with usual pathogenic therapy (24 children in the group) had 17 children with low IgA concentration, 3 with medium IgA concentration and 4 with high concentration. Low IgA concentration during the course of disease in the comparative group remained almost the same: 11,7±0,2 at the beginning and 10,9±0,4 mg/l during recovery. In the event of medium and high primary IgA concentration significant drop has been observed during the period of recovery, which 4-7 times gets ahead of results from comparative group. In the event of high IgA concentration children treated with sodium succinate were likely to have lower amount of IgA concentration in their nasal phlegm but nevertheless such reduction has been less expressed as for children of comparative group (64,4±6,0 mg/l and 45,5±6,1 mg/l, р<0,05).
Under impact of sodium succinate the amount of T – lymphocytes significantly increased not only in the group of uncomplicated ARVI, but also in development of complications (from 889±39 to 1217±57, р<0,05). Significant increase of the amount of B – cells has been observed only in the event of complicated ARVI for children treated with sodium succinate (from 285±11 to 337±19, p<0,5). Indicators of the state of non-specific resistance for majority of patients treated with sodium succinate have almost remained the same during the course of disease. The amount of thermolabile beta – inhibitors during the period of recovery has not increased their content during the period of acute respiratory viral infection.
Increase of the level of serumal interferon has been observed during dynamics of ARVI for children treated with sodium succinate from 5,2 to 10 units/ml (р<0,05).
But as it is known concentration of interferon less than 10 units/ml is not enough for relief of clinical evidence of ARVI. Thus the present researches have proved out advisability of intake of sodium succinate as the component of complex treatment for children, suffering from ARVI.
Authors: Ivanova V.V, Kurbatova G.P, Blagoslovenskij G.S, Aksenova O.A.
Acid – natural preparation for diabetes treatment
In the case of diabetes dependent on insulin, hyperglycemia goes together with other metabolic disorders: hyperlipidemy, hyperlactatemia, and ketosis. Decrease of carbohydrate on purpose to egest hyperglycemia is the main principle of diabetes treatment. The initial criterion of this compensation is absence of sugar in the urine and reduced glucose concentration in the blood during twenty four hours. Such correction of diabetes is carried out by choosing diet, injecting insulin, taking non-hormonic preparations, which reduces amount of sugar or its combinations in the blood.
The usage of glucose in peripheral tissues changes dependent on the concentration of free fatty acid and ketotic corpuscules in blood plasma. Any increase of fatty acid or ketotic corpuscules is a reason of suppression of glucose usage in the tissues. It is visible in the case of both normal and enlarged concentration of insulin. By contrast, injection of glucose reduces oxidation speed of fatty acid and enlarges usage of glucose in peripheries. That is to say, that reduction of fatty acid and ketonic corpuscules level in blood plasma may stimulate usage of glucose in the tissues and reduce amount of glucose in plasma.
In the course of our experiments we noticed that progress of diabetes growth for rats (after injections of alloxan) depends on process of lipid circulation a lot according to glucose concentration in the blood. It was proved that animals, which had injections of alloxan, could be divided into two groups in unison called “diabetic” and “non-diabetic”. Sure enough, injections of alloxan caused increase of glucose in the blood till the trespass of norm for some rats, for others concentration of glucose remained almost unchanged.
Table 1.
Parallel changes of glucose concentration and ketotic corpuscules in the blood of rats causing diabetes with alloxan
Dimensional rate
Groups
Control (n–18)
After injection of alloxan
diabetic (n–10)
non-diabetic (n–10)
Glucose
9,8±0,4
18,0±1,5
8,7±0,7
3-hidroxibutirate
127±6
433±66
127±14
Acetoacetate
89±6
222±32
93±10
Amount of ketonic corpuscules
213±11
651±91
212±7
Note. 40 mg/kg of alloxan was injected in proportion to 1 ml saline. Saline was injected for the control group. Three days later, concentration of indicated substances in the blood was measured. Glucose, 3-D –hidroxibutirate and acetoacetate was measured by fluorimetric fermentatic methods. For each alloxan group rats were selected in tens. Criterion for the selection was glucose concentration in the blood: under 10 mg and above 15 mg for “diabetic” and “non-diabetic” groups.
It is shown in the table that level of glucose concentration in the blood does not rise (diabetes does not develop0 when injection of alloxan does not cause increase of ketotic corpuscules in the blood. That is to say, such animals do not differ from controlled, their weight does not decrease and there is no polyury. Those animals may be ascribable to “non-diabetic” group. In other group of rats, “diabetic”, increase of glucose concentration and ketotic blood corpuscules, which are caused by alloxan, may be seen. A weight of these rats was 20% lower than a weight of control group. The former drank a lot of water and the polyury was noticed. Thus, the positive correlation between glucose and amount of ketotic corpuscules in the blood was a feature of rats of “diabetic” group. Spanish researches, who are working with Spreg-Doyli rats causing diabetes with alloxan, got similar (although not identical) results. Glucose accession into peripheral tissues, for example into skeletal muscles and erythrocytes, is a process dependent on insulin, whereas ketotic corpuscules spared over the tissues freely and are independent of insulin. Thus, the findings let to guess, that having diabetes, when permeability of cell membrane disconcerts because of lack of insulin, the amount of glucose concentration in the blood may be diminished by the reduction of concentration of ketotic corpuscules.
We have investigated an impact of various natural preparations on concentration of ketotic corpuscules in the blood, and identified that sodium succinate the most effectively reduces concentration of ketotic corpuscules. It, especially, increases when the animal is starving (Table 2). After twenty four hours of starving concentration of 3-D –hidroxibutirate in blood plasma of the rats used to rise to 1,95 mg, acetoacetate – to 0,5mg (norm approximately 0,2 and 0,05mg respectively), how ever the effect of sodium succinate used to reduce concentration of ketotic corpuscules significantly (to 1,0 and 0,38mg respectively) during the thirty minutes previous to animal death.
Table 2.
An impact of sodium succinate on the concentration of ketotic corpuscules in the blood of starving rats
Dimensional metabolites
Concentration, mg
Saline
Sodium succinate
3-O-hidroxibutirate
1,95±0,25
1,00±0,10*
Acetoacetate
0,51±0,10
0,38±0,12
Amount of ketotic corpuscules
2,44±0,33
1,39±0,19*
Note. Rats (in tens in each group) were starving twenty-four hours. Afterwards at 9hr 30min saline or sodium succinate (50mg/kg) was injected for them. After 30 min level of concentration of ketotic corpuscules was researching by the fluorimetric fermentatic method. Deviations of average value and average effective values from normal, *р<0,005, in comparison with control group, are listed.
Mechanism, through which sodium succinate may reduce concentration of ketonic corpuscules and glucose, is indeterminate. It is may be guessed, that oxidation of succinate, which may intensify metabolism in vivo, creates surplus amount of ATP, witch is sufficient for the activity of acetoacetate, lipid biosynthesis, herewith for the stimulation of glucose transmission to peripheral tissues.
We are inclined to think, that sodium succinate may be used for reduction of glucose amount in the blood after causing diabetes with alloxan for animals. Tentative experiments proved that impact of sodium succinate (for animals diabetes was caused by alloxan) reduces not only amount of ketotic corpuscules in blood plasma, but also glucose concentration (more effectively).
Impact of sodium succinate on glucose and ketotic corpuscules concentration in rat blood (diabetes for them was caused by alloxan).
Metabolites concentration in the blood
Saline
Sodium succinate
Glucose
30±4
16,4±4,2
3-O-hidroxibutirate
886±85
530±89
Acetoacetate
369±63
298±60
Amount of ketonic corpuscules, мкМ
1250±142
824±204
Note. Alloxan was injected for the rats in 40mg/kg. Three days later, sodium succinate (50mg/kg) was injected for one group or rats, saline – for other group. After half-an-hour all animals were decapitated and metabolites, which are described in table 1) found.
Deviations of average value and average effective values from normal were listed.
In contradistinction to insulin, such unnatural agents like sulphacarbamide may lower glucose amount in plasma without its entrance into tissues stimulation and without utilization in the tissues. In that case the application of preparations, this reduces glucose concentration in the blood through reduction of ketotic corpuscules concentration, stimulation of glucose transmission to cells and its utilization in peripheral tissues without inhibition of gluconeogenesis, may be beneficial.
Authors: E. A Kosenka, J.G. Kaminskij, M.N. Kondrasheva.
The Institute of Theoretic and Experimental Biophysics
Use of Amber Acid and other components of energy balance for treatment of oncological cases
German physiologist Otto Warburg set up the hypothesis, which claims that the cause of cancer can be found in the absence of oxygen. It happens due to various reasons, and is considered to be the first step towards cellular disorders. Those disorders may become malignant. In the case of permanent insufficiency of oxygen one part of the cells perishes, another part adapts to the newly formed conditions. The latter part of the cells causes a great activity of glycolysis and like so it fills energy deficit. According to Warburg’s mind, respiration disorder is the basis of re-formation of healthy cells to cancerous. Such a disorder may emerge in consequence of various reasons. The most important of them is the disorder of oxygen-supply.
Nevertheless, not every respiration disorder causes cancer. These are the conditions that cause malignant disorders:
· Respiration disorder must be reversible, i.e. it must remain after the extraction of agent, which caused the disorder.
· Respiration disorder is not supposed to be such as to cause cells losses, according to Warburg, a sudden loss of oxygen or high concentration of toxic are more dangerous than low and permanent influence of these factors.
· Respiration disorder must remain during the cell division, i.e. it becomes genetically descendible.
· Respiration disorder cannot have influence on glycolysis.
· Respiration disorder should not rebound on cellular respiration.
In that case, respiration disorder may become carcinogenic.
The conversion of normal cells into cancerous comes along in two stages. In the first stage, after the occurence of respiration disorder, long period of the struggle for existence of cell begins. It is true to think that the only latent period of cancer formation is none other than period, during which the selection of cells happens and the glycolysis of the tissue intensifies, after the respiration disorder. Respiration disorder may be associated with the decrease number of mitochondrions in the cell.
Warburg thought that respiration of cancerous cells is not just simply weak, but weak in comparison with really intensive aerobic glycolysis. Thus, the respiration of cancerous cells becomes inferior with regard to quality (stopping of respiration, oxidation and phosphorilation). Warburg’s theory was criticized more than once. Till now the question, weather disorder of mitochondrions may be a reason of the conversion of normal cell into cancerous, is open. The main goal of this work is the testing of the state cells mitochondrions and search for the influence on cancerous cells. This, in turn, lets heal the owner of those cells, carrier of cancer.
There were exams on three types of respiration of human’s cancerous cells and ultra-structures: specifically, ovarian cancer (OV), stomach cancer (CaVe) and mesenchymoma of connective tissue (Sa19). It is proven that great activity of glucose and weak respiration of the OV cells comes to the top. A number of mitochondrions, compared together with normal cells, decreases there. According to their external look, they were similar to mitochondrions, which are affected by the unhealthy agent for the respiration – cyanide: mitochondrions were 2 – 3 times longer. CaVe and Sa19 were breathing, though glucose was increased in them also. Populations of mitochondrions were heterogeneous in Sa19 cells: a lot of swollen mitochondrions. Majority of organoids were in, known as, mediate (with the optimum amount of energy) state. Small, newly formed mitochondrions are also observed. The facts suggest that activity of mitochondrions population in Sa19 cell is high. The look of CaVe cells and their mitochondrions is similar to Sa19, but the former are not so active and pro-mitochondrions can be seen only after irradiation of the tissue.
Table 1
Rapidity of respiration, rates of conjugation of mitochondrions and amount of ATP in cells of Sa19 mitochondrions.
Substratum of oxidation
V3
V4
АDP/О
Control of respiration (by Chance)
Alpha-ketoglutarate
2,50±0,03
1,75±0,04
2,1±0,1
2,0±0,3
Amber Acid
2,00±0,05
0,76±0,06
1,8±0,2
2,6±0,1
Table 2Amount of ATP in cells of Sa19 mitochondrions
Amount of endogenous ATP (mg/mg protein)
Amount of ATP after addition of synthesized ADP (mg/mg protein)
10,1±2,1
362,6±32,4
As shown in the Table 1 and Table 2, mitochondrions, which are abstracted from Sa19 cells, has a high rapidity of respiration, but rates of conjugation are reduced in them. Amount of endogenous ATP is lesser than in mitochondrions of normal tissue. Synthesis of ATP is fairly active.
Another set of experiments shows the rates of conjugation of cells respiration and mitochondrions. They are abstracted from rat hepathoma 22, from liver of rats, which has tumour, from laboratorially affected liver of rats (the expansion of tumor stopped for them after vaccination) also from not affected liver of rats.
Table 3
Respiration of mitochondrions, which are abstracted form various liver of rats. Substratum of oxidation – glutamine acid (5 mg/ml).
Source of mitochondrions
V3
V4
АDP/О
Control of respiration (by Chance)
1.Hepathoma (n=6)
0,85±0,05
0,42±0,06
2,1±0,2
2,0±0,1
2. Liver of rats, which has tumour (n=6)
0,30±0,03
0,12±0,02
2,2±0,3
2,5±0,2
3. Laboratorially affected liver of rats (n=3)
0,74±0,02
0,20±0,04
2,8±0,04
3,7±0,4
4.Norm (n=10)
0,49±0,01
0,15±0,02
2,6±0,1
3,3±0,2
P1-2<0,05 p3-2<0,05 p3-1<0,05 p3-4<0,05
From the Table 3 it may be noticed that rapidity of respiration of mitochondrions, which are abstracted from hepathoma, is rather higher in the liver of rats, which has tumour. Though, the rates of conjugations are almost the same. We are of the opinion that it reports about the activity of energy reactions in hepathoma (conjugation of variable mitochondrions) and their reduction in mitochondrions of liver of rats, which has tumour. High rapidity of respiration and great rates of conjugation is characteristic of mitochondrions of liver of rats affected under conditions of laboratory.
The count, basically, confirms Warburg’s hypothesis, analyzing respiration disorder of cancerous cells in a wider meaning: like respiration changes and energy reactions. Therein lies, that it is possible to try to cure cancerous cells, by regeneration normal circulatory system. Herewith it helps organism, which carries cancer, to fight against this trouble.
A group of doctors of Moscow 40th hospital followed our recommendations for usage of Amber Acid (AA) together with citric acid, alpha-ketoglutarate acid, vitamins and some other medical plants. This treatment was for volunteers. During the course of treatment patients were on almost vegetarian, anti-diabetic diet. They were taking vitamins together with special medical agents of treatment. Observation period was 5 years. Patients of 2-3 stage of disease were supervised. Treatment proceeded till and after the radical operation.
Table 4
Influence of Amber Acid (together with citric acid and 2-oxo-glutaric acid on mortality of oncological patient in a course of five years (%).
Type of tumour
Conditions of experiment
Number of patients
Mortality in a course of 5 years, %
Cervical cancer
Control
Treatment
5
10
80%
10%
Uterine myoma
Control
Treatment
10
15
50%
0%
Ovarian cancer
Control
Treatment
6
8
90%
30%
Cancer of intestine and rectum
Control
Treatment
10
10
80%
10%
Mammary cancer
Control
Treatment
10
20
60%
10%
The facts suggest that AA may be successfully used for healing oncological patients other type of results of circulatory system disorder.
Author: E.V. Kozyreva
Inst. of Biophysics, Moscow, Russian Federation
