Method of treating mycotic infections and preparation for implementation thereof
SUBSTANCE: lytic exoenzyme of Cellulomonas cellulans strain RNCIM AS-870 of molecular weight within 100 to 30 kDa and specific activity 48.3 UN/mg of protein and more is used. A preparation for treating mycotic infections contains lytic exoenzyme of Cellulomonas cellulans strain RNCIM AS-870 of molecular weight within 100 to 30 kDa and specific activity 48.3 UN/mg and more of protein and represents a topical drug.
EFFECT: lower probability of recurrences in treating mycotic infections without normal flora suppression.
10 cl, 4 dwg, 3 ex
The invention relates to medicine, namely to the treatment of mycotic infections, and can be used in particular in gynecological practice.
Among opportunistic mycotic infections candidiasis plays a leading role. About 75% of women over a lifetime has at least one episode of vaginal candidiasis (hereinafter MC), the dominant pathogen is Candida albicans. Very often VK proceeds as recurrent infection and difficult to treat. A significant problem is the recurrence of candidiasis of the mucous membranes in certain categories of patients. Some of them cause a high incidence of recurrence is defined (the recurrence of candidiasis in AIDS patients), in other cases, it remains unclear (recurrent candidal vaginitis in women with normal immunity) (see, for example, a Practical guide to anti-infective chemotherapy, n/p Listratenkov, Uselocale, Snjezana. - M., 2002). The use of antimycotic drugs are not always successful, because the chronic infection may be accompanied by formation of drug resistance. This circumstance determines the relevance of search for alternative therapies VK. One of the possible approaches to solving this problem is to study therapeutic effect of enzymes of microbial origin, without setoudeh specific for cells of the fungus morpho-functional patterns.
The known method of correcting the microflora of the vagina (EN 2131258, Smirnov and others, 10.06.1999). How is the intravaginal drug Biosporin (candles or tampons). The invention allows to adjust the microflora of the vagina when the disorders caused by pathogenic and conditionally pathogenic microorganisms of different taxonomic groups. However, the effect of this drug is mediated in any violation of the microbiota possible recurrence of candidiasis.
Known farmcampsite (EN 2007105476, Dikovisky and others, 10.09.2008) for the treatment of candidal vulvovaginitis, comprising as an active ingredient the prebiotic lactulose in the form of vaginal suppositories, as an active ingredient antimycotics (azoles: Itraconazole, terconazole, econazole; nifuratel and natamycin) in an amount of 100 mg or 500 mg, and a prebiotic in an amount of 200 mg However, it is known that in the appointment of antimycotics from the group of azoles in 20% of patients within six months of observed relapses.
The described method of complex treatment of VK (EN 2254857, kolomoitseva and others, 27.06.2005) vaginal suppositories Lexicon; drugs Multitabs, REKITSEN-RD, which can significantly improve the condition of patients in the acute phase, however, are not specific antimycotic therapy, and does not guarantee the absence of relapses within six months of the donkey treatment.
Known drug - vaginal suppositories (EN 2257197, Dulcis and others, 12.05.2004), containing in particular benzalkonium chloride, benzoic acid, witespy at a certain content of ingredients. Benzalkonium chloride bactericidal effect on a whole range of bacteria, but it is less active against pathogenic fungi, in particular Candida albicans, and benzoic acid is a pH regulator, inhibits the growth of pathogenic fungi, although not a direct impact on mushrooms, and does not complete emilinate Candida from the vagina.
There is a method of treatment of vaginal candidiasis (NZ 539019 (A), HUTMAN, 27.10.2006) using the antifungal and antibacterial drugs, such as metronidazole and miconazole nitrate, as well as probiotics and anti-virus additives. However, it is known that these drugs may develop resistance, and increased doses may cause toxic effects. On the other hand, the additive does not have a specific activity to Candida.
Also known is a method of treatment of chronic recurrent vulvovaginal candidiasis (EN 2309761, Khamaganov and others, 10.11.2007), in which three stages intrawaginalno and orally administered enterosorbent, intranasal timogen and, optionally, the vaccine "Sulcotrione", cycloferon liniment intrawaginalno, autorin In oral and eufloria the L intrawaginalno. However, the positive impact of immunomodulatory components of nonspecific resistance is not a specific effect on the causative agents of candidiasis that does not guarantee full protection from relapse.
There is a method of treatment of mycotic infections, using enzymes with lytic effect on cells (US 4062941, Davies, 13.12.1977 - the closest equivalent). The complex political eksperimentov was extracted from strains Corpinus comatus, Licoperdon depressum or Licoperdon pyriforme. Enzymes identified by laminarinase (chitinase, chitobiase). However, these enzymes destroy the component of the cell wall of the fungus - chitin, but not affect bananowy layer, which defines the basic structure of the walls of the fungus.
The present invention is a method of treating mycotic infections and preparation for its implementation, which allow you to have a more specific impact directly on the Candida albicans without the oppression of the normal microflora, since the point of application of the drug are bananowy and fibrillar layers of the cell wall of fungi. This results in the irreversible loss of Candida virulence properties and viability, which causes no recurrence.
Patent-pending method for the treatment of mycotic infections using as active substances political eksperimenta in f is RME funds for local use is characterized by they use political assofermet strain Cellulomonas cellulans PMBC AC-870 having a molecular weight in the range from 100 to 30 kDa and the specific activity of not less than 48,3 U/mg protein.
The method can be characterized by the fact that the mentioned assofermet represents litigate, as well as the fact that when intravaginal and rectal administration mentioned assofermet used in a dose of 10-50 IU once a day for 5-7 days.
The method can be characterized and the fact that the local application mentioned assofermet used in the amount of 2-10 UNITS per 1 g of biologically acceptable carrier, mainly in the form of ointment or gel, 1 times a day for 5-7 days.
The method can be characterized also by the fact that additionally given the protease inhibitor contrical, which is injected together with the aforementioned eksperimenta in quantities of 500-1000 Acre one dose of the latter.
Drug for the treatment of mycotic infections using as active substances political eksperimenta in the form of drugs for local application is characterized by the fact that they use political assofermet strain Cellulomonas cellulans PMBC AC-870 having a molecular weight in the range from 100 to 30 kDa and the specific activity of not less than 48,3 U/mg protein, as well as the fact that the mentioned assofermet represents litigate.
Preparation for local, intra the original and rectal contains 10-50 UNITS mentioned eksperimenta on a single pharmaceutical form, mainly suppository, tampon or sanitary pad.
The drug may be characterized in that it contains the above-mentioned assofermet in the amount of 2.0 to 10.0 UNITS per 1.0 g of biologically acceptable carrier, and made in the form of ointment or gel, and that contains the protease inhibitor contrical in quantities of 500-1000 Acre per single dose mentioned mentioned political eksperimenta.
The technical result - the reduction of relapse in the treatment of mycotic infections, increased antifungal activity without inhibition of normal microflora. This is ensured due to the impact of the mentioned eksperimenta as bananowy layer of the cell wall of Candida albicans and fibrillar layer, followed by its removal. Additional technical result is a synergistic effect when used together referred to political eksperimenta and kontrikala: fungi of the genus Candida formed the developed surface, however, does not exhibit pathogenic properties.
The invention is illustrated in the graphs, in which figure 1 is presented the dynamic growth of the culture-producer and changes in proteolytic activity of the culture liquid (QL); figure 2 - changes in the activity of litigate in the dynamics of the cultivation of Cellulomonas cellulans; figure 3, 4 - clearance vagina against yeast-like fungi under the influence if icasi at a dose of 2 IU/ml and 10 IU/ml, respectively.
As the active substance use political assofermet - litigate strain Cellulomonas cellulans PMBC AC-870. Before seeding on mattresses culture .cellulans stabilized by creating a cold shock (exposure inoculum for 20 minutes at a temperature of +4°C). Further, the strain is cultivated at a constant Shuttle-programming for 2.5-3 days at 37°C on a modified heart-brain broth (SMB) with the addition of dead Baker's yeast in the amount of 9.5 to 10.5 g per 1 l of the medium as an inducer of litigate. The dynamics of the growth was monitored by optical density (OD). It is established that the activity of litigati increased during exponential growth of culture and reached a maximum in the stationary phase, and then decreased. The optimal period effective cultivation is 65-72 hours, because during this period there is the greatest accumulation of the desired product (1, 2).
At the end of the cultivation QOL in a volume of 1.5 liters was centrifuged at 3000 rpm for 15 min and passed through a sterilizing filter with a pore size of 0.02 μm. Next, QOL has been diafiltration on the cells of the firm "AMICON" using membranes HM-300 HM-100 and PM-20. Each of the fractions was concentrated to a volume of 20 ml, and then determined the activity litigate. It is established that the activity of the enzyme away the evidence in the fractions, containing biopolymers with a molecular mass of more than 100 kDa and 300 kDa, but was fully manifested in the interval from 100 to 30 kDa. The obtained parameters are consistent with the characteristics described in the literature. After washing and concentration of this fraction specific activity of the final product is not less than 48,3 U/mg protein, which is more than 2 times higher than the specific activity of the commercial preparation "Lyticase" company Sigma (20 U/mg protein), adopted by us as standard.
In animals was evaluated acute toxicity of the drug, to which 0.4 ml of a solution containing 117 mg protein, was administered to 10 mice in the dorsal vein of the tail. Animals within 10 days remain healthy, from which it was concluded that the drug is not toxic in the doses that are hundreds of times higher than therapeutic.
The action received eksperimenta - litigate manifests itself in the form of lysis of the fungal cell. Lytic effect is to decrease the number of viable fungi and depends on the dose of enzyme used in the experiment. When processing suspension .albicans litigate with an activity of 10 UNITS, the decrease of the optical density of the suspension to 53% relative to control within 1 hour. Dose-dependent effect is observed when evaluating the results of seed remaining viable cells. Both processes are described by two regression equations, reflecting C the dependence of lytic effect of the dose of the enzyme:
u(x)=-10,5497856·ln(x)+67,1981181, with a standard deviation equal 5,128; where: y is the amount of decrease in optical density of the suspension of fungi under the influence of litigate, compared with control; x - dose litigate;
U(x)=-11.6411733·ln(x)+83.5368459, with a standard deviation equal to 5,945; where: Y is the percentage of viable cells relative to control; x - dose litigate.
Equations allow to calculate the 50%effect of litigati: 5,0 ED - in terms of culture lysis, 7,8 ED - cytostatic effect.
Studies have shown that remaining after exposure to litigate viable mushrooms lose their virulence, which provides for the development of infectious process. This is due to the decrease in adhesion of fungi to vaginal epithelial cells of the vagina and, therefore, decrease their colonizing ability. The impact of litigati on the surface structure of the cell wall of fungi by destroying manambolo and fibrillar layers increases about 1.5 times the seizure of their phagocytes and contributes to the activation of intracellular digestion. So, as a result of the research showed that after exposure of native mushrooms with macrophages was digested 41,0±0.4% of cells. As a result of processing litigate this figure increased up to 48.0±1,0%. The data obtained indicate that the digestive phase, Fugazi the concern is more efficient in case of damaging the cell wall lytic action of eksperimenta litigate. Experiments with isolated macrophages were allowed to establish that the effectiveness of the phagocytic process is manifested in the development of complete phagocytosis. Reduced virulence of Candida under the action of litigate due to removal of fibrillar layer and destruction mannopova layer in the cell walls of fungi leads to loss of adhesion and to increase the efficiency of phagocytosis by macrophages. In addition, it was found that the antimycotic action litigate can be enhanced by the introduction of the drug "Contrical". The drug "Contrical" (Aprotinin/APROTININ), an inhibitor of proteolytic enzymes and proteases of the blood plasma is typically used for cupping and treatment of pancreatitis (manufacturer AWD pharma GmbH & Co, Germany). The applicant was first revealed previously unknown property kontrikala to inhibit the production of enzymes aggression aspartyl-proteinases from Candida. Inhibition by contrical aspartyl-proteinase prevents the implementation of the invasive properties of the agent that blocks the transition in the pathogenic filamentous form, keeping in yeast form, and contributes to the elimination of the pathogen. Contrical enter in the quantity 500-1000 Acre together with literati, one dose of the latter. Patented formula contains a single track to litigate and contrical. Evidence of synergistic antifungal E. the reaction of two drugs is the mechanism, detected in vitro. Litigate provides a lytic effect on fungi and contrical blocks the enzymes aggression aspartyl-proteinase from Candida, preserved viability.
The study of therapeutic action litigate conducted on the model of vaginal candidiasis (VC) using a clinical strain of Candida albicans No. 40, which was isolated from the vagina of pregnant women diagnosed with vaginal candidiasis. The indicated strain were injected at 20 μl intrawaginalno at a dose of 108CFU/ml, against the background of dysbiosis caused by antibiotics and artificially induced estrus by hormone Messalina. After elimination of the pathogen from the vagina provoked a relapse of the infection by re-treatment of animals with antibiotics and hormone without the introduction of the culture of the pathogen. Monitoring of the course of infection was carried out according to two indicators of contamination: the number of CFU in 1 μl of vaginal discharge and clearance calculated by the formula: K=(C0-Ct)/S0·100%, where: K - ground clearance (purification)0the number of fungi in 1 μl of vaginal discharge mice in the initial time after infection; Ctthe number of fungi in 1 μl of vaginal discharge after a certain period of time.
At the height of the manifestation of relapse in animals from the experimental group with an interval of 2 days intrawaginalno centuries the Dili to litigate. Control sanitation in experimental and control groups was carried out by seeding mushrooms. Two series of experiments were conducted using different doses of the enzyme: 2 and 10 IU/ml (figure 3, 4). Provocation marked with letters A (an antibiotic) and M (the hormone mesalen).
From these experiments, shown in figure 3 (1-5 days - an antibiotic; day 5 - infection .albicans; 36-40 days antibiotic; 43-72 days of litigata), it follows that after the first introduction litigate is a significant reduction in contamination of the vagina Candida compared to control.
When using litigate at a dose of 10 IU/ml (figure 4) reorganization completed on 8 days earlier than at the dose of 2 IU/ml figure 4 data of experiments: 1-5 days - antibiotic and hormone; day 5 - infection .albicans; 38-42 days - antibiotic and hormone (meslin); 47-73 days of litigate; 76-80 days - antibiotic and hormone. When provocation candidiasis antibiotics and hormones repeated relapses did not develop that is proof of a complete reorganization of the vagina under the influence of litigate.
When determining the dose came from experimental models of vaginal candidiasis. Calculated, which allows to extrapolate the experimental data obtained in vitro and in vivo in animals at therapeutic doses treatment for humans. Proceeded as follows. The amount of vaginal mouse - 25 ál (25.10-6l). Therapeutic therapeutic dose, derived in experiments is e, was 2-10 IU/ml for 7 days. The amount of vaginal women 0,05 l, Respectively, extrapolating the dose volume vagina women received the specified range of values of 10-50 IU (p.3 formula), and 2-10 IU per gram of carrier.
It should be noted that the strain .albicans No. 40 is a clinical strain isolated from a pregnant woman, the patient with vaginal candidiasis. Therefore, experiments with literati and strain .albicans No. 40 in vitro and in vivo in models of vaginal candidiasis can be considered as experimental evidence on the applicability of the drug for the treatment of mycotic infections.
Studies by electron microscopy allowed us to fix what changes undergoing cell cell strain of C.albicans under the action of litigate. The chemical composition of the cell wall presents: chitin, glucan and mannan, with clearly defined fibrillar layer. Clarified that even if the destruction mannopova layer does not occur, there is a noticeable his thinning, and also leads to the destruction of the surface fibrillar layer and its removal. Around Candida visible area of the enlightenment, in some places there is a rupture of the cell wall, with glucan and chitin layers remain unchanged.
The result of this degradation is the change in the adhesive properties of fungi of the genus to which ndida: reduced their adhesion on the surface of target cells (vaginal epithelial), but it does not change adhesive activity against members of the normal microflora of the vagina (lactobacilli). The recommended dose of litigati much lower than is required for lysis of the cells. This allows to reduce the drug load with preservation of therapeutic effect.
In the proposed drug is used of litigate, the initial activity of which is not less than 48,3 U/mg protein. Specified political assofermet you can enter in different quantities in the composition of the different pharmaceutical forms of the drug to achieve the activity 10-50 IU per dose.
The preparation of litigate can be used for local, vaginal or rectal application in the form of an aqueous solution obtained by stationary or industrial technology. The solvent may be used sabove-certified saline, pH 7,2.
The preparation of litigate may also optionally contain a filler, usually used in the manufacture of the relevant prescription forms. In the manufacture of suppositories or candles as a filler is used, for example, confectionery fat, lanolin, cocoa butter, gel, aluminum hydroxide, paraffin, etc. Upon receipt of the ointment or gel vaseline, lanolin, witepsol, sodium alginate, methylcellulose, methocel, hispagel, GPMC etc. the Drug can also be lio is ilizarova.
The invention is illustrated by the following examples.
Example 1. Receiving liquid medication litigate. When industrial technology cultivation of a strain of Cellulomonas cellulans PMBC AC-870 is carried out in the reactor/fermenter with a nutrient medium for culturing at a temperature of 35-37°C for 10-24 hours prior to culture in the stationary phase of growth. After incubation, the culture fluid sephirot and by fractionation on the cartridges is isolated and concentrated extracellular biopolymer with a molecular weight in the range from 100 to 30 kDa. Activity litigate determined by lysis of yeast fungi of the genus Sacharomyces, and received the drug in the value of specific activity of at least 48,3 U/mg protein considered suitable for subsequent use and poured into vials with the introduction of the known stabilizing additives.
When the treatment liquid drug in a number containing a single dose (for example, 5 ml of buffered saline solution, pH of 7.2, containing 50 IU litigate) put on a tampon or pad. Used topically, rectally or intrawaginalno.
Example 2. Receive preparation in the form of suppository. Obtained in example 1 liquid preparation of dehydrated by spray or vacuum drying plant, combined with fillers (confectionery fat, lanolin, paraffin, vicasolum etc.) number is the number, provide not less than 10-50 IU activity litigate one suppository. So, to prepare 100 suppositories weighing 5 g each, must be entered in 500 g of filler to 103.5 mg litigate (with source activity 48,3 U/mg protein). If suppositories contain contrical, it is administered simultaneously with litigate in accordance with the calculation: one suppository - 500-1000 Acre. Next suppositories form on a special installation.
We studied the stability of the 8 experimental production batches of product. As shown by the results obtained immediately after the formation of suppositories activity litigate is not less than 40 UNITS, and after 12 months of storage were not recorded activity at least 10 UNITS on one suppository.
Example 3. Receiving the drug in the form of ointment or gel. Obtained in example 1 liquid preparation of dehydrated by spray or vacuum drying plant, combined with ointment (vaseline, lanolin, etc) or gel base (methylcellulose, sodium alginate and the like) in an amount to provide not less than 2-10 IU activity litigate per gram basis. So, for preparation of ointment (gel) require 100 g basis to enter 20.7 mg litigate (at initial activity for at least 48,3 U/mg protein). If the ointment (gel) contains contrical, it is administered simultaneously with litigate in accordance with the calculations of the volume: one gram of ointment (gel) 100-200 Acre. Further ointment (gel) Packed in tubes or bottles.
We studied the stability of the 6 experimental production batches of product. As seen from the obtained results, immediately after cooking activity litigate is at least 10 UNITS per 1 gram basis, and after 12 months of storage was not a series of drug with activity less than 2 UNITS per 1 gram basis.
The preparations obtained in the above examples, are harmless. To determine the safety of mice injected intrawaginalno the drug in a volume of 20 µl. For each variant of experience using at least 10 mice weighing 15-16, the Drug is considered harmless if all mice remain alive for 5 days of observation, and none of them identified the disease.
The preparation of Litigate characterized fungicidal activity relative to the test strain of C.albicans No. 40, which is an internal standard for determining the antimycotic action of the claimed drug.
Fungicidal action litigate was determined by the method of serial dilutions in liquid nutrient medium (Laba). From the culture of C. albicans suspension prepared containing 105microbial cells in 1 ml of the broth using bacterial optical standard. Then carried out a series of dilutions of litigati (containing from 10 to 5120 ED) in meat-peptone broth, 2 ml, all ol the tag was added to 0.2 ml of culture C.albicans. Conducted two control: cultural vitality C.albicans without the introduction of litigati and control of microbiological purity of the enzyme containing 5120 ED litigate without making culture. After 48 hours incubation in thermostat took into account the results on the presence or absence of growth. All of the tubes did the sowing of grass in 0.1 ml on a dense medium Saburo. After 24 hours of incubation in a thermostat counted the number of CFU on plates. It is established that the minimum concentration of litigate, completely inhibiting the growth and reproduction of C. albicans, amounted to 1280 UNITS, which were confirmed by sowing on a dense medium. When activity over 640 UNITS in any case carried out experience was not observed in the growth of C. albicans No. 40.
Fungistatic and fungicidal activity of the enzyme towards .albicans was determined in the experiment on the dense environment of AGW. 10 samples of 1 ml of a solution litigate in buffered physiological solution (SFR) with initial specific activity 10, 20, 40, 80, 160, 320, 640, 1280, 2560, 5120 ED was layered on Wednesday AGV and kept for 5 hours at 37°C for liquid is absorbed. In the control sample was injected SFR. After that planted 10 μl of the suspension .albicans, providing an increase of about 50 CFU in control. Analysis was performed after 24 hours of incubation in a thermostat percentage retained viability .albicans after processing, which can be the. The experiment was repeated ten times. Complete inhibition of growth of Candida was observed under the action of litigate over 640 UNITS in relation to the suspension Candida. Fungistatic action litigate began to appear with values corresponding to 10 UNITS of activity. When 40 UNITS suppressed the growth of more than 50% of the cells .albicans, with 640 UNITS - more than 98% and at 1280 IU in 100% of cases are reported complete bactericidal effect litigate.
Thus, our studies allow us to conclude that the patented method for the treatment of mycotic infections using litigate - lytic eksperimenta strain Cellulomonas cellulans PMBC AC-870, having a specific activity of not less than 48,3 U/mg protein contained in the extracellular fraction of biopolymer with a molecular weight in the range from 100 to 30 kDa and selectively destroying the basic structure of the cell wall of fungi, helps complete renovation of infectious foci. This elimination of the pathogen from the vagina so effective that the provocation of recurrent infection becomes impossible.
1. A method of treating mycotic infections using as active substances political eksperimenta in the form of funds for topical application, characterized in that use political assofermet strain Cellulomonas cellulans PMBC AC-870 having a molecular weight in the range from 100 to 30 kDa idalou activity of not less than 48,3 U/mg protein.
2. The method according to claim 1, characterized in that the said political assofermet strain Cellulomonas cellulans represents litigate.
3. The method according to claim 1 or 2, characterized in that during intravaginal and rectal administration mentioned political assofermet used in a dose of 10-50 IU once a day for 5-7 days.
4. The method according to any one of claims 1 or 2, characterized in that the local application of the mentioned political assofermet used in the amount of 2-10 UNITS per 1 g of biologically acceptable carrier, mainly in the form of ointment or gel, 1 times a day for 5-7 days.
5. The method according to any one of claims 1 or 2, characterized in that it further designate the protease inhibitor contrical, which is injected together with the aforementioned political eksperimenta in quantities of 500-1000 Acre one dose of the latter.
6. Drug for the treatment of mycotic infections in the form of funds for local use, containing as active substances political assofermet, characterized in that it contains political assofermet strain Cellulomonas cellulans PMBC AC-870 having a molecular weight in the range from 100 to 30 kDa and the specific activity of not less than 48,3 U/mg protein.
7. The preparation according to claim 6, characterized in that the said political assofermet strain Cellulomonas cellulans represents litigate.
8. The drug according to any one of p or 7, characterized t is m, for intravaginal and rectal contains 10-50 UNITS mentioned political eksperimenta strain Cellulomonas cellulans on a single pharmaceutical form, mainly suppository, tampon or sanitary pad.
9. The drug according to any one of p or 7, characterized in that it contains the mentioned political assofermet strain Cellulomonas cellulans in the amount of 2.0 to 10.0 UNITS per 1.0 g of biologically acceptable carrier and made in the form of ointment or gel.
10. The drug according to any one of p-9, characterized in that it further comprises an inhibitor of proteinases contrical in quantities of 500-1000 Acre per single dose mentioned political eksperimenta strain Cellulomonas cellulans.
SUBSTANCE: for correction of endothelial dysfunction in experiment on rats dysfunction is modelled by preliminary sensitisation of animal with staphylococcus anatoxin in doze 0.1 ml subcutaneously. Then after 24 hours in the same place as anatoxin introduced is suspension of Staphylococcus aureus subcutaneously in dose 60 billions of microbial bodies in 1 ml. After that for following generalisation of infectious agent, daily massage of injection place is carried out. At this background introduced is macrolidal medication azithromycin intragastrically in dose 30 mg/kg/day.
EFFECT: method ensures activation of endothelial dysfunction correction.
1 ex, 1 tbl
SUBSTANCE: in order to estimate resorbtion function of peritoneum time during which animals achieve stage of surgical sleep in case of intraperitoneal introduction of ethaminal is determined.
EFFECT: method allows to estimate indirectly resoptive function of peritoneum in norm and under impact of aggressive factors.
SUBSTANCE: invention relates to medicine, namely to surgery. Solution of dye is introduced in proper segmental vessel by direct injection method. Preliminarily, before introduction of dye solution, feeding artery is compressed for 2-3 minutes, dye solution is introduced more distally than compression place and after 1-2 minutes respective vein is compressed. Simultaneously area of supposed staining is influenced by source of constant magnet, placed outside of the organ surface.
EFFECT: method extends arsenal of means for marking intraorgan zones, segments of parenchymatous organs.
SUBSTANCE: invention relates to experimental medicine and deals with restoration of reducing with age potential of cell growth of tissues in old laboratory animals. For this purpose medication "Transfer-factor" is introduced to laboratory animals in dose corresponding to 1 capsule a day per 50 kg of weight.
EFFECT: method ensures activation of T-lymphocyte function and restoration of potential of cell growth of salivary gland tissues in said group of animals.
SUBSTANCE: invention relates to chemical and pharmaceutical industry, namely to creation of medication reducing hematotoxicity of cytostatics. As such medication, polysaccharides from leaves of coltsfoot (Tussilago farfara L.) are used.
EFFECT: medication extends arsenal of preparations increasing efficiency of chemical therapy by reducing henatotoxicity of cytostatics.
SUBSTANCE: invention relates to field of medicine, namely to gynecology, and deals with treatment of complicated forms of inflammatory diseases of uterine appendages. For this purpose, when performing abdominal operation from two sides between round ligament of the uterus and isthmic part of fallopian tube medicinal mixtures of the following medications are introduced: at first - lidase 16 U in 1 ml of 0.25% Novocain, after that without removing the needle prednisolone 15 mg, amikacin 0.25 mg in 1 ml of 0.25% Novocain. Drainage of small pelvis with gauze-glove pad on lateral pockets to retrouterine space and bed of remote mass is carried out. In post-operational period antimicrobial drug is introduced intravenously. Limphotropically from two sides from anterior commisure of large pudental lips introduced are medicinal mixtures of medications: first - lidase 16 U in 1 ml of 0.25% Novocain, after that without removing the needle - prednisolone 15 mg, amikacin 0.25 mg in 1 ml of 0.25% Novocain, slowly during 15 minutes. Such introduction is carried out 1 time a day for 7-8 days.
EFFECT: method ensures stimulation of interstitial humoral transport and lymphatic drainage and fast normalisation of clinical and laboratory parametres with reduction of course dose of antimicrobial drugs.
SUBSTANCE: invention relates to veterinary. Method of manufacturing biogenic stimulator, which includes 50% of biologically active mass from drone larvae, 49.7% of sodium chloride solution and 0.3% of preservative, consists in the following: in order to obtain preparation comb with alive healthy 18-22 days old drone brood is kept in refrigerator at temperature 3-4°C during 5-6 days, after that comb with brood is opened and drone larvae of the same size, light-gray colour are selected, after which drone larvae are crushed in sterile laboratory mill, diluted with sterile sodium chloride solution with concentration 0.9% in ratio 1:1 and autoclaved at temperature inside boiler 120°C and steam pressure in jacket 1.5 atmosphere for one hour, after that the mass is filtered through two layers of gause into sterile measure vessel, brought to initial volume with sterile solution of sodium chloride with concentration 0.9%, preservative (phenol) is added, preparation is poured into prepared sterile vessels observing rules of asepsis, vessels are hermetically closed, autoclaved at 120°C and steam pressure in jacket 1.5 atmosphere for 20 minutes.
EFFECT: method allows to reduce considerably terms of preparation manufacturing, increase its safety, increase storage terms Biogenic stimulator obtained in said way contributes to improvement of animal growth and development, increase of offspring preservation, reduction of feed cost per unit of body weight increase, increase of general and specific resistance of organism.
3 tbl, 6 ex
SUBSTANCE: group of inventions relates to medicine, namely to nutrition and therapy, and can be used for reduction of unfavourable effects in case of treatment with phytosterols. In accordance with invention method deals with maintenance and/or increase of bioacceptibility and/or levels in blood of at least one of circulating lipophilic vitamins, lipophilic medications, lipophilic nutrients or any lipophilic pharmaceutically active ingredient in individual who needs therapy with phytosterols by introduction of therapeutically efficient amount of mixture of phytosterol ester(s) (PS-E) and 1,3-diglyceride(s) (DAG). Edible composition in accordance with invention contains mixture of phytosterol ester(s) (PS-E) and 1,3-diglyceride(s) (DAG).
EFFECT: application of inventions allows to reduce concentration of cholesterol and triglycerides without unfavourable impact on the level of lipophilic molecules in blood.
28 cl, 5 tbl, 2 dwg, 5 ex
SUBSTANCE: invention pertains to a polymorphous modification of A N-benzyl- N'-(2,6-diisopropyl-phenyl)-N-isopropyl-malonamide, which is characterised by X-ray powder diffraction pattern with characteristic peaks at approximately 16.0, 19.6 and 28.1 for 2θ degrees.
EFFECT: obtaining a composition which can be used to make a medicinal agent for reducing secretion of sebum and treating skin disorders.
6 cl, 4 ex, 3 dwg
SUBSTANCE: invention refers to medicine, dentistry and can be used for treatment of inflammatory periodontal diseases. Luminol-dependent chemiluminescence of oral fluid is carried out. Rate of maximum flash and glow light sum is estimated. With the value of maximum flash from 0.8 to 5 c.u. (conventional units) and light sum of 1.5 to 37 c.u. an impact of magneto-infrared laser radiation is conducted. The impact is conducted with a pulse repetition rate of 80 Hz, LED radiating power of 90 mW. The impact is conducted in area of projection of the foci and in field of angle of the mandible on the same side with exposure 2 minutes per point, and then in the spinous process of third cervical vertebra during 1 minute and in oral tissues for 2 minutes. After that it is affected with magneto-infrared laser radiation with frequency of 150 Hz, LED radiating power of 50 mW. The impact is conducted on reflex points GI4 and E36 symmetrically, VC12 or MC6 and RP6 symmetrically, VC12, alternating every other day. At the same time in each session an effect is exerted on points CI 1, GI 20, E 5, V 10. The treatment course is 10 procedures. Exposure is 0.5 minutes on one point. At the maximum flash rate from 6.55 to 25 c.u. and light sum from 26 to 45 c.u. a biologically active food supplement "PropoFlan" is administered in a dose of 2 tablets 3 times a day for 1 month.
EFFECT: method provides choice of adequate treatment of inflammatory periodontal diseases taking into account the activity of oxidative processes in oral fluid.
SUBSTANCE: invention refers to medicine, namely to agents for treating gastrointestinal disorders (GID) based on probiotics, and their metabolites. There is offered a preparation containing (wt %): zeolite - 70÷80; soya flour hydrolysate - 15÷25; calcium stearate - 0.5÷1.5; lytic enzymes - 0.05÷3.5; microorganism cell lysate - the rest. The method involves microorganism cultivation, mixing of a metabolite mixture with zeolite and adjuvants, sterilisation and packing of an end product; and the prepared producing microorganism cells are treated with lytic enzymes for 2-10 hours at temperature +20-50°C. If necessary, the composition additionally contains enzymatic preparations to the preset parametres. An agent represents a preparation of broad clinical effectiveness; it is recommended in treatment regimens for the patients suffering from intestinal dysbacteriosis of various geneses.
EFFECT: when implementing the technology, constant biochemical characteristics of the composition of the declared agent are provided; living producing cells are eliminated from the end product; storage-stability of the agent is improved; preparation of the end agent is simplified due to eliminated drying stage.
8 tbl, 16 ex
SUBSTANCE: invention belongs to veterinary medicine. Method consists of ingestion of Vetom 4 once a day with colostral milk, milk or warm boiled water during morning feeding in the rate 75-100 mg/kg of body mass.
EFFECT: method allows cryptosporidiosis prevention in calves; it improves general resistance in sick animals and shortens length of treatment.
3 cl, 3 tbl
SUBSTANCE: described is adsorbing product, such as sanitary napkin, panty liners, protective means at incontinence, diaper, including adsorbing structure (4) with first and second sides, humidity-sensible additive (8) such as bacterial composition and member-carrier (10). Member-carrier (10) including first and second impenetrable for water steam layer (12, 13) of fabric, at that layers of fabric are tightened with compression (11) with formation of impenetrable for water steam volume between them (9), withholding humidity-sensible additive (8). Member-carrier (10) is situated on the first or second side of adsorbing structure (4).
EFFECT: invention allows protecting susceptible to humidity additive (8) in order to ensure that no harm can be caused to this additive or it can not be destroyed at that properties of the product are preserved.
33 cl, 12 dwg
SUBSTANCE: therapeutically effective amount of a proteosome composition containing a preparation of outer membrane protein of gram-negative bacteria.
EFFECT: reduced amyloid deposit ensured by activated microglial phagocytosis without induction of neurotoxic effects.
17 cl, 9 dwg, 4 tbl, 1 ex
FIELD: medicine, veterinary science.
SUBSTANCE: an antibacterial agent, probiotic "Sachabactisubtyl" from strains "Bacillus subtilis "ТНП-3-ДЕП"" and "Bacillus subtilis "ТНП-5-ДЕП"", adsorbed on zeolite of Honguru deposit.
EFFECT: method allows improving immunobiological eactance of an organism, fertility and preservation of young deers.
SUBSTANCE: described is adsorbing product, such as sanitary towel, everyday towel, tampon, diaper, diaper in form of underpants, protective means for incontinence for adult people, provided with at least one capsule (8), containing bacterial composition (11) in lipid phase (12), which preferably contains, at least, one strain of bacteria producing lactic acid. At least one part of capsule has minimal transversal size (a), equal to at least 2 mm. Capsule (8) preferably includes core (9), which contains bacterial composition (11) in lipid phase (12), and coating (10), preventing environment impact on core during transportation and storage.
EFFECT: adsorbing product is convenient for application, leads to efficient transfer of bacteria to region where they are located, and which can be stored during long periods of time without losing bacterial cell viability.
19 cl, 2 tbl, 1 ex, 12 dwg
SUBSTANCE: method includes bottle-feeding of EM-preparation "Baykal EM-1" to calves in mixture with infusion of herbal tea in ratio of 1:1. At the same time herbal tea contains the following components (per 100 ml of water): 0.5 g of touch-and-heal herb, 0.5 g of meadowsweet herb, 0.35 g of thousand-leaf herb, 0.35 g of stinging nettle, 0.2 g of Bergenia crassifolia rhizome, 0.2 of roots and rhizome of cowberry and 0.15 g of Scotch pine needles. Mixture is bottle-fed according to the following scheme: 1 day - twice in doses of 30 ml and 50 ml; 2 day - thrice in dose of 100 ml; further - five times a day in dose of 100 ml until clinical recovery.
EFFECT: method is highly efficient.
5 tbl, 2 ex
SUBSTANCE: there is described adsorbent product, such as a hygienic towel, a daily liner, a tampon, a diaper, a diaper panties, a protective aid for incontinence in adults, containing a bacterial composition (8) in lipid phase (9), and the specified bacterial composition contains at least one strain of bacteria producing lactic acid. The lipid phase (9) containing the specified bacterial composition (8), is applied on a carrier element (10) or therein. The carrier element, at least before the product is applied, is impenetrable to the lipid phase. The carrier element is located on the side of the adsorbent structure (4) turned to the user so that to prevent considerable transmission of the lipid phase in the adsorbent structure.
EFFECT: adsorbent product allows maintaining normal microbial flora and prevents susceptibility to microbial infections.
26 cl, 2 tbl, 7 dwg
FIELD: chemistry; biochemistry.
SUBSTANCE: method involves preparation of a stock culture through combined growth of lactic acid bacteria Enterococcus faecium B 3491, Lactobacillus plantarum B 3242, Bifidobacterium bifidum AC 1247 under submerged cultivation conditions, taken in ratio of 1:1:1 until achieving bio-titre of each bacterium of not less than 5·108 CFU/ml. Growth is carried out in a liquid medium which is a mixture of cow milk and water in ratio of 1:1. A mixture of extraction cakes-aromatic herbs is prepared from cumin seeds, coriander seeds and parsley herbs after CO2 extraction with the following ratio of components, wt %: extraction cake of cumin seeds 20-40, extraction cake of coriander seeds 30-50, extraction cake of parsley 30. The obtained mixture of extraction cakes is inoculated with the stock culture in ratio of 1:1 and incubated at 35-40°C for 4-6 days to obtain not less than 5·109 CFU/g in a semihumid preparation. The obtained semihumid preparation is mixed with the obtained mixture of extraction cakes until attaining moisture content of 8-12% in the ready preparation and a bio-titre of each microorganism of not less than 5·108 CFU/g.
EFFECT: increased safety and survival rate of young growth and increased body weight gain.
SUBSTANCE: method of selective preparation of 7α-hydroxyandrostens with the help of floccus, includes transformation of Δ5 -androstenes with the help of mycelium Curvularia lunata VKPM F-981 separated and washed from medium. At the same time substrate is used in the form of microchips, or in the complex with cyclodextrins.
EFFECT: yield of end 7-hydroxyproducts - 60-90%.
2 cl, 5 ex
FIELD: veterinary science.
SUBSTANCE: it is suggested to apply sulfatetrine M and probiotic ocarine for calves by the following dosage-temporal scheme: sulfatetrine M at the dosage of 0.2-0.4 g/kg body weight starting since the second colostral feeding, and then at a 12-h-long interval for 7 d; ocarine - at the dosage of 8x106 microbial cells once daily for nocturnal period for 10 d. The suggested combination enables to considerably decrease side effects, provides immunocorrection of the main affected immunity links, for example, prophylactic efficiency of the method corresponds to above 90% at 100% safety of calves.
EFFECT: higher natural body resistance.
2 ex, 4 tbl