Effects of MPEP on avoidance learning in rats.

In recent years metabotropic glutamate receptors (mGluRs) have received considerable attention as a potential target for psychotropic drugs, but their influence on learning and memory is still unclear. The aim of the present study was to examine whether intraperitoneal (i.p.) administration of selective mGluR5 antagonist 2-methyl-6-(phenylethynyl)-pyrydine (MPEP), injected prior to, immediately after or 30 min after training, affects acquisition and/or retrieval of the inhibitory step-down and active shuttle-box avoidance in rats. Our results indicate that 5 or 10 mg/kg i.p. MPEP in all tested groups impaired memory consolidation of step-down training without affecting acquisition and had no effect on learning and retention in shuttle-box trained rats. The data are in agreement with the statement that mGluR5s may contribute very little and task-dependently to the actual acquisition of new information, but memory formation, appears to require mGluR5s through modulation of consolidation and/or recall.

Perinatal period cytokines related to increased risk of future allergy development.

Testing of cytokine levels in colostrum, cord blood and amniotic fluid of healthy and allergic mothers and their newborns (using protein microarrays and quantitative analysis by ELISA) revealed differences in the levels of IL-5, IL-10, TGF-beta, TNF-alpha, EGF and eotaxin between healthy and allergic groups. Significantly higher concentration of IL-5 and IL-10 in the colostrum of allergic mothers and cord blood of their children and also tendency to a higher level of IL-4 found at allergic mothers and their children (but without statistical significance) indicate a bias to T(H)2 response in this group. The higher level of TGF-beta in the colostrum of healthy mothers should be involved in beneficial immunological tuning of their children including enhanced IgA formation and better intestine maturation. In amniotic fluid, concentration of TGF-beta was higher in children of allergic mothers. A significantly higher level of EGF was proved in the colostrum of healthy mothers and in cord blood of their children in comparison with allergic group. EGF deficiency in the allergic group could impair or delay intestine maturation and support thus allergy development.

Intratracheal and intranasal immunization with ovalbumin conjugated with Bacillus firmus as a carrier in mice.

Inactivated Bacillus firmus (BF), G+ nonpathogenic bacterium of the external environment, was coupled to ovalbumin (OVA) and used in immunization experiments as antigen carrier. Balb/c mice were immunized thrice intra-tracheally and intra-nasally with conjugates of OVA and BF. Surprisingly, administration of OVA-BF conjugates inhibited anti-OVA IgG response in both sera and mucosal secretions if compared to an exposure to OVA alone. The suppression of antigen-specific antibody production was accompanied by promotion of TH1 phenotype.

Immunostimulatory effect of Bacillus firmus on mouse lymphocytes.

Bacillus firmus (a Gram-positive nonpathogenic and harmless bacterium), was shown to be a strong polyclonal activator of mouse B lymphocytes as estimated by ELISA testing of Ig concentrations in culture supernatants after incubation of BALB/c mouse splenocytes with inactivated bacillus. Synthesis of all main Ig classes and all IgG subclasses was stimulated in vitro, the considerable effect on IgA formation being the most interesting feature. B cell stimulation was T cell dependent, as was demonstrated by the effect of B. firmus on all Ig isotypes and by comparison of lymphocyte response of nu/nu mice and heterozygous nu/+ mice. The effect of B. firmus on splenocyte proliferation was stimulatory or suppressive depending on the dose of the bacterium. Increased synthesis of IFN-gamma and IL-10 (detected by ELISA in splenocyte culture supernatants) showed probable stimulation of Th1 and Th2 subpopulations. Considering the stimulatory effect on IgA formation and macrophage stimulation, B. firmus seems to be a prospective mucosal adjuvant and/or probiotic.

Stability of some novel thymidine, 5-bromo-2'-deoxyuridine and 3'-azido-2'-3'-dideoxythymidine analogues.

In the search of new prodrugs effective against herpes simplex virus series of thymidine, 5-bromo-2'-deoxyuridine esters with amino acid and peptide chains and 3'-azido-2',3'-dideoxythymidine derivatives have been synthesized and evaluated for antiviral activity. The chemical stability of some of them containing different residues was studied at pH 1 and 7.4 and temperature of 37 degrees C. An HPLC method was developed for quantification of the unchanged ester concentration. It was proved that esters with simple aliphatic straight side chain (containing alanyl-, glycyl-, or glycyl-glycyl-glycyl-residues) are relatively stable both at acidic and neutral media, 37 degrees C. Some of them undergo negligible hydrolysis with half lifes ranging between 6 and 23 h. In contrast, more complex esters with branched side chain (valyl-), with phenyl residue (phenylalanyl-), as well as containing thiazol ring are rather unstable especially at acidic conditions and undergo rapid hydrolysis resulting in the respective chemical precursor. The stability of the former group esters outlines them as suitable candidates for prodrugs: with higher lipophilicity facilitating po absorption, satisfying chemical stability and possibility to release the active moiety following enzymatic hydrolysis.

Synthesis of thiazole, imidazole and oxazole containing amino acids for peptide backbone modification.

Novel 5-ring heterocyclic building blocks are synthesized. These can be incorporated into analogs of peptide antibiotics such as microcin B17, which is a potent DNA-gyrase inhibitor that exhibits eight thiazole and oxazole moieties. In particular, the syntheses of imidazole and bisoxazole amino acids as novel peptidomimetics are reported, this includes a new procedure for the oxidative conversion of the intermediates oxazoline, imidazoline as well as oxazole-oxazoline into the corresponding heteroaromatic compounds. A mixture of 1,8-diazabicyclo-[5.4.0.]-undec-7-ene carbon tetrachloride/acetonitrile and pyridine proved to be a very effective and mild agent.

Synthesis and anti-virus activity of some nucleosides analogues.

New 3'-, 5'-, 5-bromo-2'-deoxyuridine (3a-g) and 3'-, 5'-thymidine (4a-i) analogues with amino acid and peptide residues were synthesized and evaluated for antiviral activity. The influence of long peptide chains, essential amino acids and the effect of this structural modification on the antiviral activity has been also reported. Three 5-bromo-2'-deoxyuridine derivatives containing glycyl-, glycyl-glycyl- and glycyl-glycyl-glycyl- residues (3a, 3b, 3c) showed a strong activity against the herpes virus PsRV and a moderate one vs. HSV-1. The corresponding thymidine analogues were considerably less effective, and only compounds 4d and 4h showed a borderline effect against PsRV.

Is the identification of antibodies against the nervous tissue an indicator of brain injury?

Using a modification of the ELISA method, auto-antibodies against the own nervous tissue have been identified in the serum of laboratory rats. The prevalence of the IgM class of antibodies suggests their physiological significance. Antibody levels are higher in females than in males. Brain hypoxic injury brings about a shift in the spectrum of antibodies towards the IgG class. It may thus serve as an indicator of brain impairment caused by a lack of oxygen.

Effect of Bacillus firmus on antibody formation after mucosal and parenteral immunization in mice.

Immunostimulatory properties of B. firmus, a nontoxic, nonpathogenic G + bacterium of external environment, were described previously. Antiinfectious and antitumor activity, macrophage activation and strong polyclonal stimulation of B lymphocytes were proved in human, mice and rats. The adjuvant effect of B. firmus on specific antibody response to ovalbumin in BALB/c mice is the topic of the present study. Against our expectation, B. firmus exerts more suppressive than stimulatory effect on specific antibody response. Formolized B. firmus decreased anti-ovalbumin response after subcutaneous immunization and only slightly increased serum antibodies after intraperitoneal immunization. After mucosal immunization, both oral and rectal, ovalbumin itself did not cause a significant systemic response but induced IgA anti-ovalbumin response in the intestine. B. firmus applied together with ovalbumin increased systemic serum response but absolutely eliminated intestinal response. The rectal route of antigen administration has been found less convenient because of less precise dosing of antigen in this mode of immunization.

Interaction of virulent and attenuated tick-borne encephalitis virus strains in ticks and a tick cell line.

An interference between a thermosensitive (ts) mutant and the wild-type (wt) of tick-borne encephalitis (TBE) virus in Ixodes ricinus L. and Rhipicephalus appendiculatus (Neumann) ticks is reported. I. ricinus females were dually infected by a parenteral inoculation of ts and wt strains at 10-day interval. Interference was demonstrated by the lowered ability of wt virus to replicate in ticks previously infected by ts virus. The wt virus was demonstrated in only 30% of the ticks; the average virus titre was lowered by 2.1 log10 compared with the control group, which was infected with the wt virus only. The oral infection of R. appendiculatus ticks with the same viruses also revealed an interference with the growth of the superinfecting wt virus. While in the control group all the ticks became infected, in the dually infected group the wt virus was found in only 50% of the ticks. However, when the ticks were infected orally with ts virus and superinfected parenterally with the wt virus, no interference was observed. In a R. appendiculatus-derived cell line persistently infected with the ts virus (100% of the cells), a partial inhibition of the growth of the superinfecting wt virus was observed. The ts virus retained its thermosensitive phenotype throughout the persistent infection of both the ticks and the tick cell line.

[DNA, RNA and protein synthesis in hepatitis A virus-infected cells]. / Sintez DNK, RNK i belkov v kletkakh, zarazhennykh virusom gepatita A.

Synthesis of DNA, RNA, and protein macromolecules was observed in cells infected with a strain of HA virus which had undergone 11 passages in the same culture. At 24-hour intervals the infected and uninfected cells were pulse-labeled with 1 microCi/cc 3H-thymidine, 3H-uridine, and 14C-valine. In 6-9 days after cell inoculation, inhibition of DNA, RNA, and protein synthesis was observed, being most marked in DNA synthesis. The synthesis of these macromolecules increases on the 10-13th day which coincides with virus antigen accumulation in the cell cytoplasm.

[Proton transport during nerve stimulation]. / Transport protona v nerve pri vozbuzhdenii.

A decrease of external pH during rhythmic excitation of the nerve of frog and squid was investigated. The level of pH was dependent on frequency excitation, concentration of sodium potassium and hydrogen ions was changed after ouabain, 2,4-dinitrophenol, tetraethylammonium and tetrodotoxin effect. The mechanism of proton transport was discussed in relation to Na-channel function.

[Mechanism of activation of Na,K-ATPase in nerve fibers during rhythmic excitation]. / O mekhanizme aktivatsii Na,K-ATFazy v nervnykh voloknakh pri provedenii ritmicheskogo vozbuzhdeniia.

The activation mechanism of Na,K-ATPase in nerve fibres after rhythmic excitation was studied. 3H-ouabain binding to a nerve was found to depend on the frequency of rhythmic excitation. The maximum of 3H-ouabain binding to a nerve crab was at 10 imp/sec. Rhythmic excitation was found not to change Na,K-ATPase affinity to ouabain, but appeared to increase the concentration of ouabain-sensitive sites in the nerve membrane. Transformation of inactive forms of the enzyme into active ones was supposed to be a possible cause of greater 3H-ouabain binding to the nerve during rhythmic exitation.