Safety of the Drug Prospekta: a Preclinical Study of Acute and Repeated Dose Toxicity.

General toxic effect of the drug Prospekta (modified affinity purified antibodies to the brain-specific S100 protein) was studied on mature male and female mice and rats: acute toxicity with double intragastric and intraperitoneal administration of the maximum permissible doses at a 2-h interval, repeated dose toxicity with intragastric administration of the maximum permissible and close to therapeutic doses for 6 months. No lethal and toxic effects on animals were observed, including no toxic effects on vital systems, i.e., CNS and cardiovascular system, as well systems with the functions that may be temporarily disrupted (excretory and digestive systems). All the differences between animals of the experimental and control groups varied within the physiological range. It can be concluded that the drug produces no general toxic effect on laboratory animals.

Analysis of General Toxicity of Ergoferon.

For antibody-based drugs, it is important and relevant to study their toxic effects, which can often become limiting when prescribing this type of therapy. General toxicity of antiviral drug Ergoferon based on technologically processed antibodies was studied on sexually mature animals. Analysis of acute toxicity showed the absence of lethal outcomes when the drug was administered to adult rats at the maximum tolerated doses. In a study of repeated dose toxicity, no adverse effects of the drug were detected.

Spatial Memory in the Progeny of Rats Subjected to Different Types of Experimental Preeclampsia.

Spatial memory was studied in 2-month-old offspring of rats subjected to different types of experimental preeclampsia (replacement of drinking water with 1.8% NaCl from day 1 to 21 of gestation or intraperitoneal administration of non-selective NO-synthase inhibitor L-NAME to pregnant rats in a daily dose of 25 mg/kg for 7 days on gestation days 14-20). Spatial memory was evaluated in an elevated 8-arm radial maze. Both types of experimental preeclampsia impaired spatial (long-term and short-term) memory and can be used in the development of drugs correcting negative effects of this pregnancy complication on memory.

Preclinical Toxicological Study of Release-Active Preparations for Prediction of Their Pharmacological Activity and Side Effects.

We studied chronic toxicity of a few release-active preparations: Dietressa (release-active preparation of affinity-purified antibodies to type 1 cannabinoid receptor), Divasa (releaseactive preparation containing a combination of affinity-purified antibodies to brain-specific S-100 protein and endothelial NO-synthase), Cardostin (release-active preparation containing a combination of affinity-purified antibodies to C-terminal fragment of angiotensin II type 1 receptor and endothelial NO-synthase), and Bation (release-active preparation containing a combination of affinity-purified antibodies to IFN-γ and CD4). We evaluated not only side and toxic effects, but also the relationships between these effects and pharmacological activities of the preparations. The data of preclinical toxicological studies of the release-active preparations can be used for prediction of their pharmacological activity.

[Study of Mental Functions Offspring of Rats with Experimental Preeclampsia in Postnatal Ontogenesis].

The postnatal development of offspring's mental functions from rats with experimental preeclampsia caused by the replacement of drinking water by 1.8% sodium chloride solution from the 1st day of pregnancy until delivery was studied. Animals born by females with complicated pregnancy have a high level of anxiety, de- creased, motor, orienting-investigative activity, cognitive impairment in postnatal ontogenesis, compared with the offspring from rats of the control group. This model can be used in research related to the evaluation of animals born to females with the EP, as well as to search for new, effective agents for the purpose-of correcting deviations in nental development in children born to mothers with preeclampsia.

Comparison of physical development and rate of formation sensory-motor reflexes offspring of rats with different experimental model of preeclampsia.

Summary: A comparative study of the physical development and the rate of formation of sensory-motor reflexes offspring of rats with experimental preeclampsia (EP) was carried out. In the first experimental group EP was modeled intraperitoneal conduct of L-NAME at a dose of 25 mg/kg from 14 to 21 days of gestation, the second experimental group - the replacement of drinking water by 1.8% sodium chloride solution for the entire period of gestation. In the offspring of both groups, there was a delay of physical development, which was reflected in the later timing of the hair coat development, incisor eruption, pinna detachment as compared to the pups in the control group. It also noted the gap in the formation of sensory-motor reflexes and vestibular reactions. This was manifested in the delayed appearance of the olfactory response, auditory sensitivity, later performing tests «righting reflex¼, «negative geotaxis¼, «aerial righting reflex¼, «cliff avoidance¼, «horizontal wire test¼, «raising the head and forelegs¼, «supporting their bodies on hind legs¼ as compared to the indices of the pups of the female rats with an uncomplicated pregnancy. The most pronounced lag in postnatal development was observed in the offspring of rats with EP, which instead of drinking water was prepared 1.8% sodium chloride during the entire period of gestation. The purpose: To make a comparative study of the impact of ADMA-like preeclampsia (PE) and preeclampsia modeled by the replacement of drinking water consumed by female rats during gestation with 1.8% NaCl solution on the physical development and the rate of the maturation of sensory motor reflexes of their offspring. Methods: The study was performed on three groups of pregnant female rats aged 3-4 months whose original weight was 210-250 g and their pups. They were divided into three groups: 1: Control group including female rats with an uncomplicated pregnancy (n = 6) and their pups (n = 49); 2. Experimental group 1 - pregnant female rats with PE (n = 6) induced by intraabdominal introduction of L-NAME at a dose of 25 mg/kg from 14 to 21 day of gestation (ADMA-like preeclampsia) and their offspring (n = 35); 3. Experimental group 2 - pregnant female rats with PE (n = 6) modeled by the replacement of drinking throughout gestation with 1.8% NaCl solution and their pups. When studying the physical development of the pups we considered the terms of pinna detachment, hair coat development, incisor eruption and the time when they began to open their eyes. To estimate the rate of the maturation of sensory motor reflexes and motor coordination of the pups of the rats with PE we analyzed the time when they started to support their bodies on hind legs, lift their bodies off the floor, crawl, raise their head and forelegs, show the aerial righting reflex and the righting reflex, negative geotaxis, reactions to auditory and olfactory stimuli as well as the time they managed to stay on the horizontal wire. Results: The pups of the female rats with PE of both experimental groups were found to have later pinna detachment, incisor eruption and hair coat development as compared to the indices of the control group. In addition, the offspring of the experimental groups demonstrated a delay in the performance of the following tests: «righting reflex¼, «negative geotaxis¼, «aerial righting reflex¼, «cliff avoidance¼, «horizontal wire test¼, «raising the head and forelegs¼, «supporting their bodies on hind legs¼, «reaction to an olfactory stimulus¼ and «reaction to an auditory stimulus¼ as compared to the indices of the pups of the female rats with an uncomplicated pregnancy. Conclusion: PE induced by the introduction of ADMA to pregnant female rats and by the replacement of drinking water consumed by female rats during gestation with 1.8% NaCl solution causes a delay in physical development, maturation of sensory motor reflexes and vestibular reactions in their offspring.

Experimental study of the efficiency of Dietressa preparation on body weight reduction in mice feeding high-fat ration.

We studied the efficiency of Dietressa on body weight reduction in C57Bl/6 male mice feeding standard high-fat ration (24%). After 5-month daily intragastric administration of Dietressa, body weight gain was the lowest in comparison with other groups and did not differ from that in mice receiving the reference substance sibutramine for 5 months. In contrast to sibutramine, Dietressa did not increase motor activity of animals in the open field test and produced no anorectic effect. The mean body weight gain per each 1000 kcal of consumed food in the group of animals receiving Dietressa was lower than in the control group and mice receiving sibutramine.

[Comparative study of the effects of new neuroactive amino acid derivatives on the postnatal development of the rat's offspring with experimental preeclampsia].

BACKGROUND: Our aim was to investigate the effect of derivatives of GABA and glutamate on the postnatal development of the offspring of rats with experimental preeclampsia. METHODS: The experiments were performed on 35 albino female rats aged 5-7 months, weighing 220-240 g, and their offspring in the amount of 284 individuals. Experimental preeclampsia was modeled by replacing the drinking water by 1.8% NaCl solution to pregnant females from 7 to 21 days ofgestation. Glutamic acid--compound RSPU-135 at a dose of26mg/kg, GABA derivative - compound RSPU-242 at a dose of 23 mg/kg and the reference drug sulodexide in a dose of 30 mg/kg administered to female orally daily, since the 7th day of gestation prior to delivery. Evaluated the physical development of offspring, sensory-motor reflexes, mental functions. RESULTS: It was found that the experimental preeclampsia causes a delay in physical development and maturation of sensory-motor reflexes in the offspring, as indicated by the later periods of eruption of the incisors and eye opening, response to the emergence of audio and olfactory stimuli, forming vestibular stability and coordination of movements compared to pups from females with physiological pregnancy. Offspring from females with experimental preeclampsia were noted for lagging behind in mental development, as evidenced by the decline of the orienting-exploratory activity, learning and memory, increase of anxiety level. Compound RSPU-135, to a greater extent, improves physical development, increases the rate of maturation of sensory-motor reflexes, RSPU-242--stimulation of cognitive functions, keeping the memory trace, orienting-exploratory, spontaneous locomotor activity, and reduce of anxiety level. CONCLUSION: The neuroactive amino acid derivatives limit the negative effects of experimental preeclampsia on the offspring.

[Gravidaprotective action of phenibut in experimental pre-eclampsia].

It was established that the replacement of drinking water by 1.8% NaCl solution in female rats during pregnancy causes experimental pre-eclampsia (EP), as evidenced by an increase in the blood pressure, proteinuria, and edema in the control group as compared to pregnant female rats with normal drinking regime. Animals with EP exhibited disturbance of vasodilating endothelial function, microcirculation disorder, and increased coagulation and thrombogenic potential of blood. In addition, the group with EP showed evidence of the activation of lipid peroxidation (LPO) due to lower activity of antioxidant enzymes. Daily oral administration ofphenibut (25 mg/kg) in female rats with EP during pregnancy prevents the increase in blood pressure and the severity of proteinuria and edemation. Phenibut improves the vasodilator and antithrombotic endothelial functions, increases uterine blood flow, improves microcirculation, limits LPO, and increases the activity of antioxidant enzymes.

Study of the absolute bioavailability of citrocard, a new GABA derivative.

The main pharmacokinetic parameters attest to short elimination half-life and mean retention time of a single citrocard molecule. The average rate of plasma concentration decrease of the compound determined small area under the pharmacokinetic curve. Steady-state distribution volume was low and only slightly surpassed the volume of extracellular body fluids in rat, which indicated moderate capacity of citrocard to distribution and accumulation in the tissues, which is seen from low systemic clearance (Cl) despite the quick elimination of the compound. Absolute bioavailability was 64%.

[Hepatoduodenal circulation and excretion of the new GABA derivative citrocard].

Pharmacokinetic investigation of a new gamma-aminobutyric acid (GABA) derivative cirtocard showed that, upon the intravenous introduction, the drug is determined in high concentrations in organs of elimination--the liver and kidneys. The tissue accessibility amounts to 1.341 for the liver and 4.053 for the kidneys and the separation factor is 1.041 for the liver and 4.486 for the kidneys. The study of drug excretion showed that cirtocard is determined in the urine for 48 h, its nephritic clearance being 0.047 L/h and extra-nephritic clearance, 0.33 L/h. For the unchanged substance, a large significance ofhepatoduodenal circulation is low probable, since no more than 1 - 2% of the introduced dose was isolated with bile over entire experiment. It is established that the removal of the unchanged substance does not exceed 10% of the introduced dose. There is high probability of hepatoduodenal circulation and excretion of the preparation in the form of metabolites.

Experimental study of the effects of Dietressa, a new weight-reducing drug.

The capacity of a new drug containing ultra-low doses of antibodies to cannabinoid receptor type 1 (Dietressa) to reduce body weight gain in mice on a high-calorie diet was evaluated, possible mechanisms of drug action were analyzed, and its safety (abuse potential in the reaction of self-stimulation) was evaluated. Dietressa was not inferior to sibutramine in reducing body weight gain in mice and exhibited no abuse potential.

[The mode of spot test of plague and pseudotuberculosis agents mix cultures].

It is supposed to implement the polymerase chain reaction with mix of two pairs of "chromosome" primers "vlm12for'/"IS216rev" and "JS for"/JSrev" species-specific for Y. pestis and Y. psdtbc correspondingly in spot test of plague and pseudotuberculosis. The additional immunodiagnostics is applied to find defective and full-fledged on plague bacteria F1-antigen synthesis in the volume agglomeration reaction and paragglutination with diagnosticums of plasmid-dependent F1 antigen and chromosome FV determined antigen. This mode is characterized by more effectiveness and lesser labor intensiveness. It accelerates the detection of the mix of two agents in bacterial inoculations, organs bioassays and other materials. Also this mode facilitates the analysis of mix cultures of agents of plague and pseudotuberculosis, obtained in "mixed" nidi, the component identification, differentiation and isolation of pure growths of both Yersinia.

Some aspects of toxicological activity of kardosten after chronic 6-month administration to rats.

Chronic treatment of rats with kardosten for 6 months had a positive effect on some ECG values and behavior without disordering the hepatorenal functions. All effects of the drug observed during a course of treatment were leveled and the parameters reached the basal values within 2 weeks after drug discontinuation, this confirming its safety.

[Comparison of efficacy of tests for differentiation of typical and atypical strains of Yersinia pestis and Yersinia pseudotuberculosis].

AIM: To characterize species specificity of officially recommended tests for differentiation of Yersiniapestis and Yersinia pseudotuberculosis and propose additional tests allowing for more accurate identification. MATERIALS AND METHODS: Natural, laboratory and typical strains oftwo Yersinia species were studied using microbiological, molecular and biochemical methods. For PCR species-specific primers complementary to certain fragments of chromosomal DNA of each species as well as to several plasmid genes of Y. pestis were used. RESULTS: It was shown that such attributes of Y. pestis as form of colonies, fermentation ofrhamnose, melibiose and urea, susceptibility to diagnostic phages, nutritional requirements could be lost in pestis bacterial species or detected in pseudotuberculosis species. Such attribute as mobility as well as positive result of CoA-reaction on fraction V antigen are more reliable. CONCLUSION: Guaranteed differentiation of typical and changed according to differential tests strains is provided only by PCR-analysis with primers vlml2for/ISrev216 and JS respectively, which are homologous to certain chromosome fragments of one of two Yersinia species.

[Search for primers on the basis of Yersinia pestis chromosomal DNA for effective PCR identification of typical and atypical plague pathogen strains].

The authors present the results of a comparative appraisal of a set of species-specific primers in the polymerase chain reaction (PCR) identification of typical and atypical plague pathogen strains. A hundred and seventy-three strains with a species specificity of Y. pestis and 67 heterologous strains were used to appraise the well-known Y. pestis chromosomal DNA-based primers: "3a", "yp2769ms06", "vlml2for/ISrev216", "vlm33/ISfor1754", as well as "JS" specific to Y. pseudotuberculosis". In some plague bacterial strains, the DNA sequences complementary to the primers "3a" and "yp2769ms06" were not found. All conceivable plague pathogen strains were detectable only with the primers "vlml2for/ISrev216" and "vlm33/ISfor1754". The primers "JS" recognized only the strains of pseudotuberculosis causative agent. The DNA of other microorganisms failed to react with none species of primers. For the effective detection and differentiation of typical and atypical Y. pestis strains, the authors propose PCR using a few pairs of primers with the merits of the primers of the group "vlm" and 'JS".

[Methods of diagnostics and differentiation of the plague agent: intraspecies differentiation of Yersinia pestis. Part II].

State Research Institute for Plague Control, Rostov-on-Don In the second part of the review aspects of intraspecies differentiation of the plague agent are discussed. Special emphasis is placed on the necessity of more precise definition of taxonomic position of plague agent isolates considering genotypical characteristics, data on their selective virulence, and evolutionary origin of the genus Yersinia.

[Correction of cardiotoxic effect of chronic alcoholic intoxication by GABA-ergic compounds].

Toxic effect of 50% ethanol solution (8 g/kg) on the heart appeared as diminishment of contractile properties of the myocardium. Structural GABA-analogues - citrocard (50 mg/kg), phenibut (50 mg/kg) and pyracetam (200 mg/kg) prevented injurious action of alcohol. This was evidenced by higher indices of contraction and relaxation velocity, and left ventricular pressure in intervention groups of animals compared with controls during functional tests with volume and maximal isometric load.

[Effect of standardized solution of brine mineral bischofit (MgCL2.6H2O) on the antenatal development of rat fetus].

A standard solution of mineral bischofit has low toxicity and produces a dose-dependent effect on the antenatal development of rat fetuses. Bischofit in a dose of 0.01 ml/kg and 0.1 ml/kg (daily requirement for magnesium ions) increased the index of fertility and the quality of embryonal development in rats. The index of fertility was reduced and fetal skeleton formation was inhibited after the administration of bischofit in a dose of 1.0 ml/kg.

[Approaches to species-specific typing of double mixed cultures comprising pseudotuberculosis bacteria and atypical plague bacillus strains].

The species relevance of atypical Yersinia strains was determined by various microbiological, immunological, and genetic (including polymerase chain reaction) tests. These strains were shown to represent mixed cultures of Y. pseudotuberculosis serovariant O1b and Y. pestis var antiqua. Identification-resistant cells with atypical properties and plasmid segregation were found in the populations of Y. pestis strains. Analysis of different diagnostic tests revealed the most reliable ones selected for the identification of atypical Y. pestis strains with unstable genome.