Multipurpose multifunctional high-efficiency alkali solution composition, preparation thereof and use as nonspecific immunostimulant

FIELD: applied immunology.

SUBSTANCE: composition contains, wt parts: borax decahydrate1-25, sodium thiosulfate pentahydrate 10-5-10-4, potassium carbonate 30-150, refined sugar 30-200, and water 100-200 per 100 wt parts of sodium metasilicate pentahydrate. In addition to its capability of improving resistance to diseases, body weight increase, productivity of agricultural plants, quality of crop, and ripening term (harvest time), composition according to invention possesses nonspecific immunostimulating activity, including production of antibodies and enhancement of immunity through activation of immunocytes thereby maximally strengthening vaccination effect regarding diseases caused by malignant neoplasm viruses.

EFFECT: increased assortment of immunostimulating agents.

10 cl, 11 dwg, 12 ex

 

The technical field to which the invention relates.

The invention relates to a multi-purpose high-performance composition of the alkaline solution, its preparation and application as a nonspecific immunostimulant. More specifically the present invention relates to the composition of the alkaline solution, which contains, first of all, metasilicate sodium (pentahydrate) and has non-specific immunostimulating activities, such as production of antibodies and strengthening of the immune system that is caused by activation of immune cells, thereby increasing until the maximum effect of vaccination against induced virus malignant diseases, to its preparation and application as an immunostimulant.

Description of the prior art

The functional activity of alkaline components in the body attracted great interest of researchers since the beginning of the 20th century. Currently, extensive studies have established that alkaline components in the body increase the rate of ionization of potassium and sodium, thereby increasing the ability of blood to the purification that leads to the clearance of blood, reduces fatigue and slows down the aging process. One of the compounds of alkaline solutions is described in Korean patent No. 128110, issued in the name of the authors of the present invention. This solution contains 10-18 miscast. si is ikat sodium, 0,1-0,5 miscast. the hyperoxic sodium, 5-10 miscast. potassium carbonate, 1 miscast. sodium carbonate, 10-18 miscast. refined sugar and 0.1 to 3.0 miscast. the silver thiosulfate in water. Currently, the specified composition is used for post-processing of fibrous materials and fermentation of feed in agriculture due to its effectiveness in the long-wave infrared radiation, antibacterial activity and deodorizing properties. However, the drawback of the composition is the complexity of its receipt and storage for a long time.

Meanwhile, the number of antibiotic in the world increases every year. However, when the number of input antibiotics increases the number caused by side effects. For example, for the treatment of patients who have abused the use of antibiotics, need a higher dose of antibiotics, since they have acquired resistant to them. In addition, misuse and abuse of antibiotics has led to the emergence of superbugs that have a high level of resistance (fully resistant to existing antibiotics. One of the ways to reduce the use of antibiotics is the General strengthening of the immune system that increases the effectiveness of vaccination. For example, despecialized doctors are of great interest nonspecific Immunostimulants (hereinafter designated as "NIS"), which induce the increase of the immune response of the organism to external pathogens, and at the present time all over the world conducted extensive research on the development of the NIS.

Japanese researchers have found that a substance extracted from the edible mushroom (Lentinus edoddes), has anti-cancer effect. In addition, already a hundred years ago NIS were found in bacteria. Currently being actively studied as the role of NIS in relation to the formation of antibodies and the induction of cytokines, and increased immune activity in their use. For example, it was found that the extract of the cell membrane Norcardia opaca induces activation of macrophages obtained from the peritoneal cavity of mice (Barot-Ciobaru and others, 1987). Found that substance RU41740 derived from Klebsiella pneumoniae, KP-40 derived from Propionibacterium avidum, and QH, Typically obtained from saponaria, have valuable properties associated with the induction of cytokines and stimulation of immune cells (Bessler and others, 1997; Nimier and others, 1999; Ronnberg and others, 1997; Siwiki and others, 1998; Tewari and others, 1996). It is now established that derived from bacterial DNA CPG motifs, called motifs-Immunostimulants, have the ability to induce the expression in immune cells, IL-6, IL-12, IL-18 and IFN-γ (Bohle and others, 1999; Klinman and others, 1999; Krieg, 1999).

Problems associated with the use of these and other designed NIS, are the complexity of their on the teachings, the impossibility of long-term storage and high cost.

Summary of the invention

In connection with the known from the existing prior art problems, the present invention was based on the task of developing a composition having inhibitory activity against the proliferation of bacteria and viruses alkaline solution which can be easily prepared and stored for a long period of time and which can be used as a nonspecific immunostimulant.

Another object of the present invention is a method for preparing such a composition of the alkaline solution.

The next object of the present invention is the use of such a composition of the alkaline solution.

And another object of the present invention are methods of increasing the rate of weight gain of cattle and crops of cultivated plants.

Another object of the present invention is a method of storing agricultural, fish or meat products for a long period of time.

And another object of the present invention is non-toxic non-specific immune with the anti-cancer action.

Brief description of drawings

The above and other objects, features and other advantages of the present invention is more explained in detail below with reference to the accompanying drawings, showing:

figure 1 is a graph of time - dependent changes of CD4+T-lymphocytes in pigs from group to feed which was added BARODON®and the control group;

figure 2 is a graph of time - dependent changes in the number of cells in presenting MHC class II in pigs from group to feed which was added BARODON®and the control group;

figure 3 is a graph of time - dependent changes in the content of the "no T"/"no-lymphocytes (N-cells) in pigs;

figure 4 is a graph of time - dependent changes in the level of CD8+T-lymphocytes in pigs from group food which was added BARODON®and the control group;

figure 5 - histogram lymphoproliferative activity of lymphocytes isolated from the peripheral blood of pigs from group to feed which was added BARODON®and the control group in response to the stimulation by Con A (concanavalin a), PHA (phytohemagglutinin), PWM (root extract of lacunosa) and LPS (lipopolysaccharide);

figure 6 - histogram lymphoproliferative activity of lymphocytes pigs, isolated from mesenteric lymph nodes of pigs from group to feed which was added BARODON®and the control group in response to stimulation by Con A, PHA, PWM and LPS;

figure 7 is a plot of the production of cholera antibodies in animals from group to feed the cat who Roy was added BARODON ®and the control group from time to time;

on Fig - graph of changes in the muscle activity of CA-ATP-ASE in cod, which were treated BARODON®and control on the duration of storage;

figure 9 is the NMR spectrum, which shows a band of wavelengths corresponding to the peak absorption for labelled with17About tap water; and

figure 10 and 11 NMR spectra, which presents the band of wavelengths corresponding to absorption peaks for labelled with17Of water molecules in an aqueous solution containing composition of the present invention.

Detailed description of the invention

The present invention relates to NIS, which is based on the composition of the alkaline solution definite composition. The composition contains metasilicate sodium, borax, sodium thiosulfate, potassium carbonate, refined sugar, and water, optionally sodium chloride, thiosulfate silver and/or sodium molybdate.

Used according to the present invention metasilicate sodium carries five molecules of crystal water and contains silicon dioxide (SiO2) in the amount of 27.5-29.0% and a sodium oxide (Na2O) in an amount of 28.5-30.0 per cent, and the limits of the error contents of these substances are less than 2%. This compound is very stable compared to marketed liquid silicate intothree is. Metasilicate sodium, a powder or granules of white color, it is easy to weigh with great precision, convenient to store for a long period of time and transport. When dissolved in water metasilicate sodium becomes strong alkalinity. Silicon, like other components sodium metasilicate, is an important element for the growth of animals and plants.

Borax containing ten molecules of crystal water, has a specific density 1,715. Component borax boron (B) is a trace element, deficiency of which have animals and plants. The lack of soil boron fruit trees do not produce a crop, and if the fruits are formed, many of them are defective. As a rule, borax is used as a repellent for bacteria repellent for insects, hemostatic means and coupling material. When alloying with metals borax has the ability to dissolve metals. A solution of borax in water has alkaline properties, the pH value is about a 9.5. In the composition of the present invention, the amount of borax is preferably adjusted to 1-15 miscast. in terms of 100 miscast. sodium metasilicate. For example, if borax is used in the amount less than 1 miscast., its actions do not appear. On the other hand, if the borax content exceeds 15 parts is Art., it can have a toxic effect.

Containing five molecules of crystal water sodium thiosulfate is not soluble in alcohol, but dissolves in water, giving it a characteristic salty taste. The aqueous solution is neutral (pH 6.5 to 8.0). Because sodium thiosulfate chemical has the ability to dissolve silver halides or other salts of silver, it is usually used for the extraction of silver from ores. In addition, sodium thiosulfate can be used to remove chlorine and heavy metals. According to the present invention, the sodium thiosulfate is used in an amount of about 10-5-10-4miscast. in terms of 100 miscast. sodium metasilicate. For example, when using sodium thiosulfate in an amount below the lower limit, it is possible to receive no further action. On the other hand, the use of sodium thiosulfate in an amount exceeding the upper limit causes the sedimentation of tissue Sa in animals, which leads to their excitation and bowel movements.

Potassium carbonate soluble in water and its solution has a strong alkalinity (pH of 11.6). Like sodium, potassium is an essential element for the body, playing an important role in metabolism and blood circulation. For example, for the prevention of hypertension and diabetes mellitus think the m appropriate balance between potassium and sodium in vivo. According to the present invention, the preferred amount of potassium carbonate is 30-150 miscast. in terms of 100 miscast. sodium metasilicate. When handling animals or plants, for example, the amount of potassium carbonate that is outside the specified range, it is possible to upset the balance of sodium/potassium in vivo.

In the composition of the present invention refined sugar prevents the recombination of ionized inorganic substances, thereby stabilizing the composition. In addition, refined sugar turns inorganic substances into organic matter type associated with sugar forms, and also improves adhesion or adsorption capacity of the composition. These features refined sugar may exercise in its application in the amount of 30-200 miscast. in terms of 100 miscast. sodium metasilicate. The water content in the composition is 100 to 200 wt. frequent. Sodium chloride, which is not a required component of the composition of the present invention, used as a source of sodium to regulate the balance of sodium/potassium. As for the silver thiosulfate, it is often used to suppress the synthesis of ethylene in plants or to facilitate differentiation of plants. In aqueous solution of silver thiosulfate at a low concentration of S2About32-form [Ag(S2About32-)2]3-while a high concentration of S2About32-form [Ag2(S2About32-)6]10-. According to the present invention, the silver thiosulfate like sodium thiosulfate exists in the polyvalent anionic form, and its activity is associated with cell differentiation. According to the present invention the sodium molybdate is used as the source of molybdenum, representing the element that is virtually absent in animals and plants. All optional compounds are used in amounts of 10-1miscast. or less based on 100 miscast. sodium metasilicate.

After complete dissolution of the components of the composition according to the present invention painted in ivory color. The composition has no odor and is non-toxic, and in addition, it is stable and has a specific viscosity of 1.43-1.50 is constant pH value of 13 and a viscosity of from 61,0 to 239,0. In particular, even after Hcl treatment, the composition of the present invention poorly curing or changes in pH. With the help of various experiments conducted over a long period of time, demonstrated that the composition according to the present invention can increase the speed of weight gain of cattle and crops of cultivated plants, as well as the is a very nice NIS as for plants, and for the animals.

The composition of the present invention can be fed to animals, for example, in parallel with the feed, mixed with food, or after fermentation in the stern, or after dilution with water. After absorbing composition leads to a very high level of immunity in cattle. For example, the composition of the present invention effectively prevents epidemic diarrhoea of pigs (EMF) and cholera swine - diseases that pose a serious problem for pig production on an industrial basis. In addition, we discovered that the composition is effective for the prevention and treatment giving the highest mortality rate of typhoid birds - a disease that causes serious damage to the poultry carried out on an industrial basis. In the case of dairy cows when included in food composition decreased the number of somatic cells per unit volume of milk, which is a measure of the quality of milk. Other effects of the composition of the present invention to cattle include enhancing growth and improving the quality of the meat. Found that the barns where the animals, feed them add the composition of the present invention, have less odor than the barns where animals are kept in the usual stern.

The composition according to the present invention has favorable impacts is imposed on the plants at its introduction in mixtures with fertilizers or after dilution with water, including facilitating the germination and growth, increase disease resistance, higher yields of cultivated plants, improving the quality of harvest, etc.

Below the invention is illustrated in the examples, not limiting its scope.

Example obtain 1

The purified water (500 kg), was added, keeping the temperature at the level of 60-80°, metasilicate sodium pentahydrate, 300 kg), borax (carbohydrate, 35 kg), sodium thiosulfate (0.01 kg), sodium chloride (1 kg) and potassium carbonate (150 kg) and was stirred for 3 hours to dissolve. In the homogeneous solution was added refined sugar (450 kg), and then was stirred for 4 h, receiving an alkaline solution with a pH of 13 (hereinafter designated as "BARODON®-1").

Example of getting 2

In the BARODON®- 1 (1,436 kg), obtained according to the example of obtaining 1, was added dropwise a solution of silver thiosulfate (0.02 kg) in water (1 l)was stirred and kept at a temperature of about 50°C for 4 h in the incubator. The resulting solution was designated "BARODON®- 2".

Example for the preparation of 3

In purified water (5 l), keeping the temperature at 100°C, was added with stirring sodium molybdate (Na2MoO4·2H2Oh, 0.3 kg)to give a colorless solution with no odor, which then was added dropwise BARODON®- 1, was stirred and kept at pace is the atur 50° C for 4 h in the incubator. The resulting solution was designated "BARODON®- 3".

Example 4

Using 6 kg of borax and 300 kg of potassium carbonate using a method similar to the method described in example 1 was obtained alkaline solution. To the alkaline solution (1,557 kg) was added dropwise a solution of silver thiosulfate (0.02 kg) in water (1 l)was stirred and kept at 50°C for 4 h in the incubator. The resulting alkaline solution was designated "BARODON®- 4".

Example of getting 5

Using 150 kg of refined sugar and the method described in example 4, was obtained alkaline solution with a relative low viscosity (hereinafter designated "BARODON®- 5").

Example 1

Efficiency in rice cultivation

Evaluated BARODON® - 1 - BARODON® - 5 to enhance immunity and promote growth of crops of rice in the rice fields.

Each BARODON® - 1 - BARODON® - 5 was diluted with 10 volumes of water, and again was diluted with water to 500 times. After immersion for 24 h in a diluted solution of rice seeds were sown in seed beds in the greenhouse. 2 days before transplantation of the rice plants from seed beds in the rice fields, young plants are sprayed with a conventional method for dilute solutions. Spraying the diluted solution was performed again two weeks before troublevania rice (heading rice).

the Growth in seed beds of young rice plants, processed composition of the present invention, was more homogeneous and strong, and they had practically no damage associated with cold weather, compared with the control group of plants that were not treated with the composition. In addition, it was found that the young rice plants treated with composition, fully rooted just three days after transplantation. In field conditions the treated rice plants were not surprised by any disease, including black leg, piricularia leaves and stem canker stems. In addition, the treated rice plants had increased resistance to lodging, and found not to have died after Typhoon, which fell 25% of the control plants.

Assessed amount received rice, the data below in table 1.

Table 1
TrackHarvest km2(the rice varieties Chuchung®)
Control575 kg
BARODON®-1655 kg
BARODON®-2690 kg
BARODON®-3710 kg
BARODON®-4680 kg
BARODON®-5670 kg

Example 2

E is the efficiency when the cultivation of the pear trees

BARODON® - 3 was mixed with 9 volumes of water and then diluted 500 times. After making pear nursery compost and organic fertilizers, the plants were sprayed with the diluted composition. After that, around the kennel was placed a fence preventing the entry of animals. About two weeks before flowering pear trees, the land was sprayed BARODON® - 2. Pear trees, fertilized compositions of the present invention, bloomed for about 4-5 days earlier and had been told to collect the fruit for approximately 15 days earlier than control plants that did not fertilize the compositions. Pears collected from treated trees had the best characteristics from the point of view of appearance, size, and measure Brix. The pears collected from plants fertilized compositions, 60% had reduced the appearance of black spots, and they were more soft in comparison with the control. On the treated trees was not detected watery pear. The dropping of fruits as a result of Typhoon with pear trees treated with compositions of the present invention, was 20% lower compared with the control. Pears collected from treated trees were approximately 15% larger compared with the control. Figure Brix for pears collected from treated trees was 13,0-15,0, which is about 12% higher than the counter in the Le. Improved the ability of the fruit to storage.

Example 3

Impact on plant growth

Diluted 16-fold solution BARODON® - 3 in water were subjected to additional breeding, the multiplicity of which are shown below in table 2, and then sprayed the leaves of the experimental plants. It was found that the treated leaves were 15.6% longer 8.7% wider and had 7-47% higher weight in the wet state compared with the control group not treated with a solution.

Table 2
The growing vessel, No.Maximum leaf width (cm)
The multiplicity of cultivation
200 times300 times400Without spraying
9thday after spraying (July 10)16,16,16,2of 5.4
26,06,36,25,9
Average6,16.26,2the 5.7
The growing vessel, No.The mass of leaves in the wet state (g/ve is marianny vessel)
The multiplicity of cultivationWithout spraying
200 times300 times400
9thday

after 2nd spray (July 10)
157,859,244,639,9
256,3of 60.542,841.5
Average57,1 (140%)59,9 (147%)43.7 (107%)40,7 (100%)

An example of obtaining 6

BARODON® - 2, prepared according to the method described in example getting 2, was diluted with multiplicity breeding 10 and 500 g of the obtained solution. Used for spraying 1 ton of feed, receiving multifunction (high) feed (indicated next BARODON®-6").

Example of getting 7

Diluted 10 times BARODON® - 2, obtained according to the method described in example getting 2 in water (10 l) together with refined sugar (3 kg), sodium chloride (1 kg) and water (75 l) was added to the feed (1 ton), after which the resulting mixture was fermentatively for 24 h under stirring, obtaining a multifunctional (high) feed (hereinafter designated "BARODON® - 7").

Example 4

A series of compositions BARODON® has been evaluated in relation to able the tee to cause weight gain and increase the immunity of animals as follows.

(1) the Ability to cause the weight gain of the pigs

For the experiment took 30 goals obtained by crossing three breeds (Yorkshire x Landrace x Durroc) hybrid fattened pigs, each age of 15 weeks (104±4 days). All of them were placed in 3 pens for pigs, each size 4×4.2 m, based on 10 pigs per pen and adapted to the new conditions for 1 week prior to the start of the experiment.

BARODON® - 6, prepared according to the method described in example 6, were included in the food groups of 10 pigs (labeled "TX-1 group") in one of the pens for pigs for 9 weeks, while in another pen for pigs a group of 10 pigs (labeled "TX-2 group) were fed a composition containing 3% BARODON® - 7, prepared according to the method described in example 7. The control group were given the same feed but without additives "BARODON®". After that, all pigs were given normal food. During the experiment, all pigs had free access to feed.

During the 6 weeks for each group was assessed by weight gain and food intake, and from these data was calculated feed efficiency (intake/weight gain). It was estimated that the average daily weight gain of the control group was 842,86 g, for the TX-1 group - 890,48 g and TX-2 group - 880,95 g, this indicator in the last two groups appeared to be improved is by 5,65% to 4.52%, respectively, compared to the control group. Average daily feed intake was 2,71 kg for the control group, 2,77 kg for the TX-1 group and 2.65 kg for the TX-2 group. Thus, the feed conversion in the control group was 3,22, for the TX-1 group calculated that the average is equal to 3.11, which means the improvement of 3.54%, and the TX-2 group feed conversion was 3,01 that means improving on 6,98%.

(2) the Taste of pork

Pork from each test group were given a taste healthy volunteers of both sexes*, and they evaluated the taste and quality of meat.

Table 3
The softness of the meatThe softness of the meat
softhardgoodaverage
Control0803
TX-16182
TX-2110100
* The number of people responding to questions after the test.

(3) the Distribution of immune cells in the peripheral blood of pigs

Using monoclonal antibodies specific against the surface of lenoci is s, and the flow cytometer, for example, supplied by the firm Dickinson Immunocytometry System, San Jose, California, USA, type FACSCalibur, in the peripheral blood of animals from TX-1 and TX-2 group and the control group was determined by the ratio of cells expressing major histocompatibility complex (MHC), and subpopulations of lymphocytes.

(3-1) Isolation of leukocytes from the peripheral blood

Blood taken from the anterior Vena cava pigs, well mixed with citrate-dextrose solution (ACD)-ethylenediaminetetraacetic acid (add) and put it on ficol-Pak (Histopaque, the company Sigma, Saint Louis, Missouri, USA). After centrifugation at 1500 rpm for 30 min received leukocytes, three times washed with phosphate buffered saline (SFR, pH of 7.2) and suspended in medium RPMI-1640 (firm GibcoBRL, Grand island, NY, USA). To estimate the number of leukocytes brought up to 1×107cells/ml, while viable cells were counted by means of the analysis of exceptions Trypanosoma blue.

(3-2) Monoclonal antibodies to assess subpopulations of leukocytes

Effects on the immune system of pigs, including populations of immune cells was determined using a monoclonal antibody with specific reactivity against cell membranes of porcine leukocytes, i.e. antigens MHC class II Rho(pork)CD2, PoCD4, PoCDS, ernestinovo IgM (sIgM), neither/nor (γδ TCR, T-cell receptor), and granulocytes and monocytes (G + M), which are listed below in table 4.

Table 4
Mat*The isotype MatMolecules**The cell type***Link
RTAIgG2aMHC class IAll nuclear cellsDavis and others (1987)
NAIgG2aMHC class IIAr-presenting cellsDavis and others(1987)
TN A5IgG2aMHC class IIAr-presenting cellsDavis and others(1987)
MSA4IgG2aPoCD2T-cellsDavis and others (1987)
RTAIgG2aPoCD4Th/i-lymphocytesDavis and others (1987)
RTWIgG2aPoCD8T c/s (T-lymphocytes, suppressor)Davis and others(1987)
Plg45 AndIgG2bSIgMIn cellsDavis and others (1987)
RTAIgG2aγδTCRN-cells Davis and others(1987)
DH59IgG1Granulocytes + monocytesGranulocytes + monocytesDavis and others(1987)
*Mat: monoclinal antibody with specific reactivity towards different subpopulations differentiation of leukocytes

** Molecules: the molecules of the various subpopulations differentiation of leukocytes pigs

***The type of cells: Cells expressing molecules

(3-3) Flow cytometry

Using flow cytometry CellQuest ratio of subpopulations of leukocytes was analyzed according to the method described by Davis and others (1990). In order to take advantage of flow cytometry with the laser beam, the cells were labeled using the indirect method with one or two fluorescent dyes, such as fluoresceinisothiocyanate (FITZ) and phycoerythrin (PV). In each well of 96-well microplate with V-shaped bottom made of 100 µl isolated from the blood of cells (cell density 1×107cells/well) and 50 μl of monoclonal antibody (concentration of 15 μg/ml) and was senzibilizirani for 30 min at 4°C, then washed three times first buffer for washing [SFR, 450 ml; ACD, 50 ml; 20% NaN35 ml of horse serum without gamma-globulin (firm GibcoBRL), 1 ml; 250mm etc, 20 ml of 0.5% phenol red, 1 ml] by centrifugation. After decantation of the supernatant at the bottom of the cell debris suspended using a mixer for tablets or vibrating mixer (firm Scientific Industries, Bohemia, NY, USA.).

In the analysis using a single dye conjugated with FITZ goat antimurine antibody IgG+IgM (firm Caltag Lab, USA), which served as secondary antibodies, diluted with magnification of 200 breeding and made of 100 µl in each well, which was suspended leukocytes. After sensitization for 30 min at 4°using centrifugation was carried out three washing by using a second buffer for washing, which had the same composition as that of the first buffer for washing, did not contain horse serum. To fix the cells in each well was added a solution of 2% SFR-formalin (38% formalin, 20 ml, SFR, 980 ml) in an amount of 200 μl.

In the analysis using two dyes for coloring cells PoCD4 (FITZ) conjugatively with PoCD8 (PV), PoCD4 (FITZ) with MHC class II (PV) and D8(FITZ) with MHC class II (Feh). In more detail, the method consists in the following: leukocytes from one of the pits was mixed with a pair of monoclonal antibodies and subjected to primary sensitization depending on the results of the analysis using one krassi is El, then washed three times with the first buffer for washing at 4°C. After this goat antibody, specific for each isotype monoclonal antibodies were added based 1.0 microgram per well for conjugated with FITZ molecules and 0.1 μg per well for conjugates with PV, and then subjected to the second sensitization for 30 min at 4°C. Processes of leaching and fixation was carried out similarly to the procedures described for analysis with a single dye.

After completion of the painting process of the cells prior to the study were stored at 4°in a dark, cool place. Using flow cytometry 2000 or more stained cells were analyzed in relation to the number of positively reacting cells. For measurement and evaluation software has been used FACScalibur and CellQuest (firm Becton Dickinson).

The results allowed us to establish that the relative content of CD4+T-lymphocytes in the peripheral blood of pigs began to rise after 3 weeks after the start of feeding animals containing BARODON® food, as demonstrated in figure 1, and on the eighth week after the start of feeding in TX-1 and TX-2 group were found significantly higher levels of CD4+T-lymphocytes compared to the control group (p<0,05). In particular, in the TX-1 group higher levels of CD4+-T-limp is Titov was maintained during the period from the 8th to the 13th week after the start of the experiment (p< 0,05).

The high level of CD8+T-lymphocytes found in the TX-2 group at 3 weeks after the start of the experiment (p<0,01), however, significant differences compared with the control group not found, starting from 8 weeks of the experiment (p<0,05), as demonstrated in figure 4.

The number of cells expressing antigens of MHC class II, mainly macrophages, TX-1 group was significantly increased compared with control to 11 weeks after the start of the experiment (p<0.05), and in the TX-2 group - to 8 the week after the beginning of the experiment, as demonstrated in figure 2.

The number of "no T"/"no-lymphocytes (N-cells) in the TX-2 group remained at a higher level, compared with the control group (p<0,01) 3 weeks after the start of the experiment, as demonstrated in figure 3, these results suggest a possible increase of both nonspecific and specific immune defense reactions. In the period between the 11th and 13th week after the start of the experiment, the number of "no"/"no-cell" remained at a significantly higher level in the TX-2-group compared with the control group (p<0,01).

Comparison groups, eating food with the addition of two different compositions BARODON®, has allowed to establish that the TX-2 group has a strong advantage compared to the TX-1 group in terms of number of cells, expre shirouma antigen MHC class II, during the period from 3rd to 8th week after the start of the experiment (p<0,05), as demonstrated in figure 2. In the TX-2 group 3 weeks after the beginning of the experiment revealed a somewhat lower number of cells expressing CD4 and CD8, in comparison with the TX-1 group (p<0,1), as demonstrated in figure 1 and 4. For the TX-2 group compared with the TX-1 group has been a noticeable increase in the number of "no T"/"no-cell" of the 13th week after the start of the experiment, as demonstrated in figure 3.

(4) Effect on the activity of blood lymphocytes and lymph nodes

To assess the activity of blood lymphocytes and lymph nodes were assessed for their proliferative response by including a [3H]-timid in porcine lymphocytes obtained from peripheral blood and mesenteric lymph nodes after stimulation concavalin (Con A), phytohemagglutinin (PHA), the root extract of lacunosa (PWM) and lipopolysaccharide (LPS).

When the PWM stimulation of lymphocytes obtained from peripheral blood TX-2 group 8 weeks after the start of feeding containing BARODON® food, found a higher level of proliferation, as evidenced by a higher stimulation index (IP) for TX group compared with the control group (p<0,05). To 11 weeks after the start of the experiment for lymphocytes isolated from peripheral blood as the TX-1 group is, and the TX-2 group found more pronounced proliferative response upon stimulation with any of the stimulant PHA, PWM and LPS compared with the response of lymphocytes isolated from peripheral blood of the control group, as evidenced by higher values of IP (p<0.01), which is shown in Fig 5.

For lymphocytes, isolated from mesenteric lymph nodes, on the 8th week after the start of the experiment were found significantly higher values of IP compared to the control group in terms of reaction as the TX-1 group, and the TX-2 group on PHA stimulation (p<0.05) and PWM (p<0,01). At 11 weeks after the start of processing in the TX-1 group were found more pronounced stimulated by Con a and PHA lymphoproliferative response compared with the control group (p<0.01)and for the TX-2 group found higher values of IP in response to the stimulation by Con A, PHA and PWM (p<0,05), which is illustrated in Fig.6.

(5) Effect on the proportion of subpopulations of CD4+- and CD8+T-lymphocytes of the spleen and lymph nodes

For analysis of the ratio of T-lymphocytes used immunohistochemical analysis. After immunological staining using the ABC method (avidin-Biotin-peroxidase complex) CD4+-, CD8+- and CD4+CD8+- dpp-T-lymphocytes in mesenteric who limfaticeskih nodes and spleen were determined using an image analyzer (Olympus, USA.). The results of the immunohistochemical analysis are shown in tables 5 and 6 below. As can be seen from table 5, while increased levels of CD4+T-lymphocytes in the spleen was found only in the TX-2 group, the content of CD8+- and CD4+CD8+T-lymphocytes significantly increased in both TX-1 and TX-2 group (p<0,001). On the other hand, in the mesenteric lymph nodes in both groups, TX-1 and TX-2 significantly increased the content of all CD4+-, CD8+- and CD4+CD8+- dpp-T-lymphocytes (p<0.01), which is shown below in table 6. In particular, higher levels of subpopulations of T-lymphocytes were found in the TX-2 group (p<0,01).

Table 5
GroupCD4+CD8+CD4+CD8+dpp
Control11±18±13±1
TX-111±111±16±1
TX-214±117±111±1
Table 6
GroupCD4+CD8+ CD4+CD8+dpp
Control32±529±210±1
TX-135±439±432±3
TX-240±447±535±4

(6) the Impact on the production of antibodies after vaccination of swine cholera vaccine

Titers of antibodies to the virus cholera swine IFA was determined after vaccination swine cholera vaccine groups of animals fed BARODON®and the control group, the food which was not added BARODON®. Significantly higher antibody titers detected three weeks after the start of feeding BARODON® the groups of animals fed BARODON® (TX-1 and TX-2) compared with the control group, and these titers remained at a high level up to 11 weeks after the start of the experiment (p<0.01), which is illustrated in Fig.7.

(7) Effect on carriers of the causative agent of typhoid birds in the absence of treatment with antibiotics

10000 of chickens infected with the causative agent of typhoid fever, for 10 weeks contained in the stern with the addition of 5 wt.% BARODON® - 7, prepared according to example getting 7. Weekly assessed their mortality, the results are presented below in table is e 7. During the experiment, infected with the causative agent of typhoid chickens introduced no antibiotics. As can be seen from table 7, in eating BARODON® group mortality did not exceed 0.3% to 15 weeks after the start of the experiment, while a large part of the additional infected with typhus control group, consisting of 10000 heads, food which was not introduced BARODON®died after only 4 days after the start of the experiment.

Table 7
Infected with typhus groupAge (weeks)Mortality over time (%)
Weeks
123456789101112131415
With the addition of the feed BARODON®270,340,360,310,370,300,280,30,270,30,40,350,330,330,320,31
The control group281,12 1,451,737,19almost all birds were killed

(8) the Impact on the number of somatic cells in milk of dairy cows suffering from mastitis

To assess the impact of BARODON® the number of somatic cells in milk of dairy cows suffering from mastitis, within two months contained in the stern with the addition of 5 wt.% BARODON® - 7, prepared according to example getting 7. Counted the number of somatic cells. The results are presented below in table 8, in which the term "liquid medicine" understand options, according to which the udder of cows rubbed the towel dampened with diluted 100 times BARODON® - 2, the term "diluted in water" understand what the cows were allowed to drink diluted 200 times BARODON® - 2, "q" denotes the number of surveyed Vimana.

Table 8
Weeks after the start of the experimentThe somatic cell count (x 1000)
Farm AndFarm B
food + liquid medication (q=10)food. (q=34)diluted in water (q=6)control (n=37)food + liquid Lekarstvo (q=16)the ORM (q=12) control (n=36)
To2,4573561,471168967619155
21,8462051,9992761,214886197
(75)(57)(135)(164)(125)(150)(127)
41,5431441,6833711,017772305
(62)(40)(114)(220)(105)(124)(196)
62,0012282,3675171,137681353
8(81)(64)(160)(307)(117)(115)(147)
1,788272845423936606210
(72)(76)(57)(251)(96)(97)(135)

Was also counted the number of main cause mastitis pathogens obtained the results presented below in table 9.

Table 9
PathogenThe number of pathogens
The group eating BARODON®Control
Before the start of the experimentAfter 4 weeksAfter 8 weeksBefore the start of the experimentAfter 4 weeksAfter 8 weeks
Susceptible sp.1176121415
Streptococci sp.221233
Gram-negative bacteria322344
Fungi/bacteria000122
Only16119182324

Example 5

We have also studied the impact of BARODON® to strengthen the human immune system. For this purpose were produced texts for children, printed on sheets of paper, which was sprayed diluted 100-fold composition BARODON® - 5, you need a kitchen is certain according to the process of obtaining 5, and the Japanese Institute of Far InfraRed Ray Application Institute (Japan Institute for the application of far infrared radiation), located in Osaka, Japan, was estimated QRS-waves. The results are presented below in table 10. For information, QRS wave number in the range from 1000 to 2000, can have a positive physical effect on the body.

Table 10
The wave number
Sprayed BARODON®paperRaw paper
Allergy170011
Optic nerve130013
The beam autonomic nerve fibers150011

Example 6

Toxicity assessment

The toxicity of the composition BARODON® evaluated through experiments on determination of acute toxicity in rats in the center of screening and toxicology of the Korean research Institute of chemical technology (Screening and Toxicology Center of the Korea Research Institute of Chemical Technology) (test No. S-700). The experiments on the evaluation of acute toxicity was performed using diluted 10 times BARODON®-4, according to the notification of the National Institute of health (National Institute of Health), No. 94-3 "Toxicity Testing Practice for Medicines" (14 APR is La 1994) and rule of the Ministry of Health (Ministry of Health and Welfare), No. 87-80 Korean Good Laboratory Practice" (October 29, 1987).

According to the results of the test toxic symptoms, including death, change of eight parameters, and other side effects were not detected neither for males nor for females rats, which were injected by the injection of a tested product, it was found that the magnitude of LD50is more than 5000 mg/kg for males and for females. This means that the composition according to the present invention is non-toxic to humans.

Example 7

Test to assess the mutagenicity

For the detection of mutagenic properties BARODON® test was carried out to identify recurrent mutations in the Centre of screening and toxicology study (Screening and Toxicology Center) of the Korean research Institute of chemical technology (Korea Research Institute of Chemical Technology) (test No. S-694) according to the notification of the National Institute of health (National Institute of Health), No. 94-3 "Toxicity Testing Practice for Medicines" (14 April 1994) and the rule of the Ministry of health (Ministry of Health and Welfare), No. 87-80 Korean Good Laboratory Practice" (October 29, 1987), capable of handling 4 strains of Salmonella (His-less mutant Salmonella typhimurium TA100/TA1535 (type of mutation is the replacement of base pairs) and ÒÀ98/TA (type of mutation shifts the reading frame) diluted 10 times BARODON®-4.

For all four studied strains of bacteria obtained negative results, suggesting that the studied product does not cause the return of the mutation with the transformation of His --negative strains in His+- positive strains. This result indicates that the composition of the present invention is safe for humans.

Example 8

Effects on cell proliferation in the fertilized egg

Immature oocytes cows were allowed to Mature in vitro for 24 h and then fertilized in vitro for 20 h Obtained fertilized eggs were placed for development for 9 days in medium containing obtained by cultivation in 500, 200, 100 and 50 times, respectively, previously diluted 10 times BARODON®-4, and control environment. Bullish VAS deferens obtained from a local slaughterhouse, the epithelial cells of the bovine oviduct (VOES) and granular cells (LC), obtained from ovaries of cows, grew with similar environments to the third generation. Using hemocytometer counted the number of viable cells.

After growing eggs, fertilized by external fertilization, crushing of eggs occurred at 60.0% of eggs in the control environments, 62.4 per cent in environments with a 500-fold dilution, to 66.3% in environments with a 200-fold dilution and 73,7% in environments with a 50-fold dilution. Estimated levels of development blastocyst embryos was 13.3% in the control environments, 20.0% in environments with a 500-fold dilution, 21.3% in environments with a 200-fold dilution, 18.4% in environments with 100-fold dilution and 11.0% in environments with a 50-fold Rabaul the tion. After culturing, the number of viable cells was 2.0×105cell type WOES and 3.2×105cells of gear type in the control environments, 2,4×105cell type WOES and 3.9×105cells of gear type in environments with a 500-fold dilution of 2.5×105cell type WOES and 3.7×105cells of gear type in environments with a 200-fold dilution, 2,6×105cell type WOES and 2.7×105cells of gear type in environments with 100-fold dilution and 2.5×105cell type WOES and 2.8×105cells of gear type in environments with a 50-fold dilution.

The above results indicate that the composition of the present invention has a positive effect on the proliferation WOES and gear, and on the proliferation and development of fertilized cells and blastocyst embryos. In particular, plasticity embryo proliferation rate increased by 50.4-60,2% when cultivated in media with 500-fold and 200-fold dilution compared with their cultivation in the control environments.

Example 9

Antitumor activity

Inhibitory activity of BARODON®-4 after its dilution in the 600-900 time was evaluated in relation to the tumor (cancer) cells of 4 types.

Cells human leukemia (Jurkat)cells of human lung carcinoma (NCI-N), human stromal cells (SW59) and cells osteogenic sarcoma human U-2 OS) were obtained from American type culture collection (American Type Culture Collection, Rockville, MD., state Marileen, USA). Each cell line was cultured in the media, which added BARODON® - 4 diluted in 600, 700, 800 and 900 times. Cells were evaluated and determined the rate of proliferation of each subculture, the number of viable cells was determined using a method based on Trypanosoma blue.

To determine the anticancer activity of healthy cells cultured in media with the same dilution of BARODON® to assess the potential cytotoxicity of BARODON®. The results presented below in table 11, indicate that dissolved environment does not inhibit the rate of cell growth of human fibroblasts and does not inhibit the proliferation of kidney cells of the Chinese hamster.

Table 11
Cell lineCulture mediumDilution BARODON®-4The number of subculturesThe cells*
CCD-27SK (healthy fibroblasts of human skin)DMEM+10% FCSControlR8Beautiful
1/900R8Beautiful
1/800R8Beautiful
1/700R8Very good
1/600P7Poor
KSS-21 (healthy kidney cells Chinese hamster)DMEM+10% FCSControlR17Beautiful
1/900R17Beautiful
1/800R17Very good
1/700R17Good
1/600P11Poor

*Painted 0,4%Trifanova blue, count produced under the microscope

The data obtained for cultures grown from tumor cells 4 types presented below in table 12.

Table 12
Cell lineCulture mediumBreeding BARODON®-4The number of passagesThe cells*
Jurkat (human leukemia)RPMI-

1640+10% FCS
controlP7beautiful
1/900P7the death of 97% in P7
1/800P7the death of 99% in P7
1/700P2the death of 99% in P2
1/600P2 100% death in P2
NCI-H69 (small cell carcinoma of the lung of a person)RPMI-1640+ 10% FCScontrolP6beautiful
1/900P6the death of 81% in P6
1/800P5death to 98% in P5
1/700P2100% death in P2
1/600P2100% death in P2
SW579 (thyroid carcinoma person)RPMI-1640+ 10% FCScontrolP10beautiful
1/900P10very good
1/800P10beautiful
1/700P10very good
1/600P9good
U-20S (osteogenic sarcomaMccoy's 5A + 15% FCScontrolP10beautiful
1/900P10beautiful
1/800P10very good
person)1/700P9the death of 39% in P2**
1/600 the deaths of 47% in P2**
* Painted 0,4%Trifanova blue, counts were made under a microscope

** 100% death after RH

As can be seen from tables 11 and 12, the composition of the present invention have inhibitory activity against human tumor cells without damaging the healthy cells. Thus, the present invention can be used for effective treatment of some types of human cancers, although the efficiency may be different depending on the tumor type.

Example 10

Impact on the preservation of freshness

Fresh cod was subjected to freezing with dry ice before transfer to the laboratory. After immersion for 10 min in one of the following solutions: solution not containing any of BARODON®, a solution containing 0.05%, 0.1% or 0.5% BARODON®-2, fish kept at 0°C for 7 days evaluating its freshness during this period.

As an indicator of the freshness of the fish was estimated muscle activity of CA-ATPase associated with the denaturation of proteins. The best results were obtained by immersion cod in 0.5%solution of BARODON®-2, the original freshness was maintained for 5 days, which can be seen from Fig.

Example 11

Effect on the activation of water

In aqueous solutions, in which BARODON® 5 Rabba who ranged in proportions 1:2800 and 1:5600 tap water, estimated17O-NMR H2O. Spectroscopic evaluation was carried out at 20°using spectrophotometer JNM-EX270 in the center of the Japanese Water Science Research Meeting, Japan, using as control the tap water. Band of wavelengths corresponding to absorption17Oh, were measured at 149,6 Hz (full width at half the peak height) for control (Fig.9), 53,6 Hz for diluted 2800 times solution (figure 10) and 54.7 Hz for diluted in 5600 time, solution (11).

The results of the spectroscopic analysis suggests that tap water is heterologous phase, due to the aggregation of various combinations of water molecules, which differ from each other by the number of bound water molecules, while containing BARODON® solutions are almost crystalline phase, in which the combination of water molecules consist of a constant minimum number of associated water molecules. In the presence of a smaller number of associated water molecules water is in a more active state, resulting in better yield and biological permeability. Because the activated water can be used for the cultivation of animals and plants, the composition of the present invention can be used as an additive to drinks or agent for treatment of waste water.

Example 12

The behavior of BARODON® when neutral is purpose

BARODON® - 4, prepared according to example 4, is strongly alkaline, pH value is 13,20. When BARODON® - 4 was neutralized with 0.1 G. of HCl, the precipitate formed on the area of contact. In addition, the composition is not detected significant changes in pH even after adding significant amounts of HCl. This behavior indicates the high reactivity of the composition of the present invention due to the fact that it is sufficiently ionized.

When diluted 1000 times the composition is neutralized HCl 0.1 N., under the microscope was not detected in any coagulates, no bubbles. In this case, the pH value has changed. Judging by the behavior of BARODON® during neutralization, it can be assumed that the application of the appropriate amount of the composition according to the present invention will not form coagulates after interaction with HCl in the stomach of humans and animals. In addition, with the introduction of the compositions of the present invention in the stomach is not detected, the generation of gas, which suggests that the application of the composition will not be problems associated with the gas.

As noted above, in the processing of animals and plants, the composition of the present invention can improve disease resistance, weight gain, yield cultural the plants, the quality of the harvest, ripening time (acquisition time). In addition, the composition according to the present invention has a non-specific immunostimulating activities, including the production of antibodies and increase immunity by activating immune cells, thereby increasing until the maximum effect of vaccination against diseases caused by viruses malignant diseases.

Taking into account inhibitory effect against some types of human cancers composition of the present invention can be used as a therapeutic or prophylactic agent.

In addition, in the stables, which contain animal feed which add to the composition of the present invention, detected less unpleasant smell of food and to a lesser extent affected by harmful insects. In addition, the composition of the present invention is an effective preservative, with the ability to maintain the freshness of food.

The present invention described above for the purpose of illustration and it should be obvious that the applied examples serve only to illustrate and not limit the scope of the invention. Based on the above descriptions, in the present invention can be made of numerous modifications and variations. Thus, it is obvious that without straying from the scope of the following formula izaberete the Oia, you can find different from the described practical embodiment of the invention.

1. The composition of the alkaline solution, which has enhanced nonspecific immunostimulatory activity, containing 100 parts by weight of sodium metasilicate formula Na2SiO3≅5H2Oh, 1-15 parts by weight of borax formula Na2In4O7≅10H2Oh, 10-5-10-4parts by weight of sodium thiosulfate formula Na2S2O3≅5H2Oh, 30-150 parts by weight of potassium carbonate, 30-200 parts by weight of refined sugar formula With12H22O11, 100-200 parts by weight of water and 10-1parts by weight or less of at least one compound selected from a range, including sodium chloride, silver thiosulfate and sodium molybdate.

2. The composition of the alkaline solution for the treatment of tumors containing 100 parts by weight of sodium metasilicate formula Na2SiO3≅5H2O, 1-15 parts by weight of borax formula Na2B4O7≅10H2O 10-5-10-4parts by weight of sodium thiosulfate formula Na2S2O3≅5H2O, 30-150 parts by weight of potassium carbonate, 30-200 parts by weight of refined sugar formula C12H22O11, 100-200 parts by weight of water and 10-1parts by weight or less of at least one compound selected from a range, including sodium chloride, silver thiosulfate and sodium molybdate.

3. The composition of the alkaline solution to enhance the immune response to BA the criteria, containing 100 parts by weight of sodium metasilicate formula Na2SiO3≅5H2O, 1-15 parts by weight of borax formula Na2B4O7≅10H2O 10-5-10-4parts by weight of sodium thiosulfate formula Na2S2O3≅5H2O, 30-150 parts by weight of potassium carbonate, 30-200 parts by weight of refined sugar formula C12H22O11, 100-200 parts by weight of water and 10-1parts by weight or less of at least one compound selected from a range, including sodium chloride, silver thiosulfate and sodium molybdate.

4. The composition of the alkaline solution to enhance the immune response to viruses containing 100 parts by weight of sodium metasilicate formula Na2SiO3≅5H2O, 1-15 parts by weight of borax formula Na2B4O7≅10H2O 10-5-10-4parts by weight of sodium thiosulfate formula Na2S2O3≅5H2Oh, 30-150 parts by weight of potassium carbonate, 30-200 parts by weight of refined sugar formula C12H22O11, 100-200 parts by weight of water and 10-1parts by weight or less of at least one compound selected from a range, including sodium chloride, silver thiosulfate and sodium molybdate.

5. The composition of the alkaline solution to enhance differentiation of fertilized eggs containing 100 parts by weight of sodium metasilicate formula Na2SiO3≅5H2Oh, 1-15 parts by weight of borax formula Na2B4O7≅10H2Oh, 10-5-10-4 parts by weight of sodium thiosulfate formula Na2S2O3≅5H2Oh, 30-150 parts by weight of potassium carbonate, 30-200 parts by weight of refined sugar formula C12H22O11, 100-200 parts by weight of water and 10-1parts by weight or less of at least one compound selected from a range, including sodium chloride, silver thiosulfate and sodium molybdate.

6. Nonspecific immunostimulant containing composition according to claim 1 as an active ingredient.

7. Therapeutic agent for the treatment of tumors containing composition according to claim 2 as an active ingredient.

8. Antibacterial agent containing the composition according to claim 3 as an active ingredient.

9. Anti-virus agent containing composition according to claim 4 as an active ingredient.

10. The amplifier differentiation fertilized eggs containing composition according to claim 5 as an active ingredient.



 

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1 tbl, 3 dwg

FIELD: veterinary medicine.

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SUBSTANCE: invention proposes applying 2,4-dichlorophenoxyacetic acid tris-(2-oxyethyl)-ammonium salt as an immunomodulating agent. The newly found properties provide the development on its base medicinal agents for treatment of inflammatory, autoimmune and lymphoproliferative diseases.

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4 tbl, 4 ex

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3 cl, 16 tbl, 9 ex

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SUBSTANCE: invention relates to preparations of vegetable origin. Invention proposes the preparation eliciting immunomodulating effect and comprising aqueous-alcoholic extracts of the following plants, mas. p. p.: glabrous licorice roots, 5.0-15.0; fenestrate Saint-John's-wort herb 4.0-16.0; dandelion roots, 4.0-16.0; sandy immortelle flowers, 3.5-14.0; senna leaves, 3.5-14.0; thyme herb, 1.0-4.0; snowdon rose (rosewort) rhizomes and roots, 0.05-0.2. Also, invention proposes the preparation comprising condensed or an aqueous-alcoholic extract of above said plants and a filling agent. The preparation can be made as tablet or capsule. The preparation normalizes the blood free radicals content and provides elevating level of T-suppressors with the simultaneous reducing amount of T-helpers in blood of patients in treatment of acute and chronic inflammatory diseases of breathing organs.

EFFECT: valuable medicinal properties of preparation.

2 cl, 3 tbl, 1 ex

FIELD: medicine, obstetrics, gynecology.

SUBSTANCE: at the background of therapy conducted one should introduce derinate immunomodulator into the body of pregnant woman additionally nasally per 1-2 drops of 0.25%-solution into each nasal canal 5-8 times daily for 3-5 d and - parenterally per 5.0 ml of 1.5%-solution once daily for 3-8 d along with preparation that improves microcirculation and along with antioxidant at a certain sequence, moreover, derinate should be introduced 30-40 min after application of microcirculation-improving preparation, and antioxidant - 20-30 min after derinate's introduction. The present innovation favors decreased edemas, decreased body weight, stabilization of Macluer-Aldrich test that in its turn enables to avoid perinatal losses, decrease the risk for the development of fetoplacental insufficiency and intrauterine fetal infection.

EFFECT: higher efficiency of therapy.

1 ex, 2 tbl

The invention relates to biopharmacology and comes to obtain the aimed at restoring metabolic processes and correction functions of the immune system

FIELD: medicine, oncology.

SUBSTANCE: method involves carrying out chemotherapy. Firstly, dose of nicotinic acid is administrated that is sufficient to initiate hyperemia of skin integuments and in appearance of first symptoms of hyperemia cytostatics are administrated by parenteral route. Method provides increasing duration of contact of cytostatics with tumor cells and micrometastasis due to inclusion of blood volumes depots in the parent state in vascular skin networks to volume of circulating blood. Invention can be used in treatment of malignant neoplasms with cutaneous and subcutaneous localization.

EFFECT: improved treatment method.

2 ex

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