[Parasitological factors impeding the transmission of the agent of babesiosis (Babesia microti) to man from the tick Ixodes persulcatus].

Based on the analysis of own and literature data, it is concluded that the following ma- in permanent system of ecologicalarasitological factors prevents the effective vector functions of the tick I. persulcatus in transmission of B. microti: lack of distinct nymphs' anthropophily; small spontaneous invasion of hungry adults; a duration of the parasitic phase in humans is insufficient to complete the sporogonic development, because victims interrupt the phase. Therefore, not excluding the possibility of sporadic babesiosis disea- ses, it can be stated that within the boundaries of a vast territory, where the taiga tick is the only potential source of infection for humans, the B. microti infection has not, and will not reach significant values in infectious pathology.

Small Heat Shock Proteins and Distal Hereditary Neuropathies.

Classification of small heat shock proteins (sHsp) is presented and processes regulated by sHsp are described. Symptoms of hereditary distal neuropathy are described and the genes whose mutations are associated with development of this congenital disease are listed. The literature data and our own results concerning physicochemical properties of HspB1 mutants associated with Charcot-Marie-Tooth disease are analyzed. Mutations of HspB1, associated with hereditary motor neuron disease, can be accompanied by change of the size of HspB1 oligomers, by decreased stability under unfavorable conditions, by changes in the interaction with protein partners, and as a rule by decrease of chaperone-like activity. The largest part of these mutations is accompanied by change of oligomer stability (that can be either increased or decreased) or by change of intermonomer interaction inside an oligomer. Data on point mutation of HspB3 associated with axonal neuropathy are presented. Data concerning point mutations of Lys141 of HspB8 and those associated with hereditary neuropathy and different forms of Charcot-Marie-Tooth disease are analyzed. It is supposed that point mutations of sHsp associated with distal neuropathies lead either to loss of function (for instance, decrease of chaperone-like activity) or to gain of harmful functions (for instance, increase of interaction with certain protein partners).

Mutations of small heat shock proteins and human congenital diseases.

The structure and properties of different members of a large family of small heat shock proteins (sHsp) playing an important role in cell homeostasis are described. Participation of the N-terminal domain in formation of large oligomers and chaperone activity of sHsp is analyzed. The structure of the α-crystallin domain of sHsp is characterized and the role of this domain in sHsp dimerization and chaperone activity is discussed. The properties of the C-terminal region of sHsp are described, and its participation in formation of large oligomers and chaperone activity are analyzed. The data from the literature on HspB1 and HspB3 mutations are presented, and involvement of these mutations in development of certain neurodegenerative diseases is discussed. Mutations of HspB4 are described and data on involvement of these mutations in development of cataract are presented. Multiple effects of HspB5 mutations are analyzed, and data are presented indicating that mutations of this protein are accompanied by development of different congenital diseases, such as cataract and different types of myopathies. The data on HspB6 and HspB8 mutations are presented, and feasible effects of these mutations on proteins structure are analyzed. Probable mechanisms underlying sHsp mutation-induced development of different congenital diseases are discussed.

[Etiological structure of complex Ixodes tickborne borreliosis natural foci of southern taiga].

89 primary isolates of B. garinii and 72 B. afzelii from different developmental phases of I. persulcatus, I. trianguliceps and form small mammalian hosts of Borrelia were obtained at an area of ca. 30 km2 located in low-mountain southern taiga forests (Perm region). The area provides home for two Borrelia species (B. garinii and B. afzeli) and their natural carrier Ixodes persulcatus. 23 isolate of B.garnii were obtained from skin biopsies and blood samples taken in patients with borreliosis. The isolates were studied by sequencing rrf(5S)-rrr(23S) spacer. The term genetic variant (genovariant) is proposed for the totality of isolates belonging to a given genetic subgroup of the concrete genospecies and having a similar nucleotide sequence of rrf(5S)-rrr(23S) spacer or other conservative genomic sequence. Genovariant is ths smallest intraspecies taxonomic unit in widespread Borrelia pathogenic for man. Several genovariants of B. garinii and B. afzelii may simultaneously occur in combined parasitic systems formed by these spirochetal agents of Ixodes tick-borne borreliosis. Such natural foci in southern taiga of the Perm region have a complicated etiological structure due to the presence of 14 genovariants of Borrelia belonging to the two above genetic subgroups. Specific genovariants occur annually but with different frequency. They are lacking in host-specificity.

[Genetic variants of Borrelia garinii, a widely spread Eurasian pathogen of ixodic tick borreliosis].

As a result of PCR-RFLP analysis and the degree of similarity between the nucleotide sequences analysis of the rrfA-rrlB intergenic spacer DNA of 227 primary isolates of Borrelia garinii and 71 isolates/ amplicons from GenBank database in different regions of Eurasia revealed significant intraspecific heterogeneity among those of Borrelia. It was shown that genospecies B. garinii had within the two genetic subgroups (20047 and NT29) 16 genetic variants, whose geography was likely to be different.

[Genotyping some Borrelia burgdorferi sensu lato isolates from Ixodes ricinus ticks in Russia and Ukraine].

The 4 Borrelia burgdorferi sensu lato isolates obtained from 1. Ricinus ticks collected in the natural foci in Russia and Ukraine, having an unusual RFLP Msel-pattern, were studied using sequencing rrfA-rrlB spacer and rrs gene. The Ir-5215 isolate from the tick collected in southern Ukraine represented recently described genospecies B. spielmanii pathogenic for humans. The three atypical isolates Ir-3519, Ir-4721, and Ir-4812 had 100% identity with the sequence of the atypical European B. burgdorferi sensu stricto strains. They constituted a subgroup of the B. burgdorferi sensu stricto on the grounds of Multilocus sequence analysis (MLSA). These data can be indicative of the genetic heterogeneity of the current group B. burgdorferi sensu stricto.

[Isolation of tick-borne borreliosis agent from blood of patients].

During spring-autumn period of 2006 Borrelia were isolated for the first time in Russia from blood of 79 patients treated in Perm City Clinical Hospital for Infectious Diseases No. 1 with diagnosis "tickborne borreliosis, manifestive form with migrating erythema, localized stage". Ten primary isolates (12.7% of total seeded samples) were obtained by seeding plasma samples on the BSK medium. Their subsequent identification by polymerase chain reaction-restriction fragments length polymorphism revealed presence of Borrelia garinii NT29 in all patients. Length of sequenced fragment of rrfA-rrlB region was 253 b.p. Seven isolates had 100% and 3 - 99.6% similarity with typical strain NT29 (L30130). Nucleotide sequences of 4 obtained isolates were deposited in GenBank database (No. AM932199 - AM932202). It was proposed that B. garinii NT29 more frequently than other Borrelia species can be an etiologic agent of tick-borne borreliosis not only in Perm region but also in whole Russia.

[Microorganisms of the order Rickettsiales in taiga tick (Ixodes persulcatus Sch.) from the Pre-Ural region].

The PCR and sequence analysis revealed DNA Ehrlichia muris, Anaplasma phagocytophilum, and Rickettsia spp. in the I. persulcatus ticks and blood samples from a patients with acute febrile illness occurring after a tick bite, registered in the seasonal peak of the tick activity of one of the highly endemic areas of Russia (Perm region). These data confirmed the validity a diagnosis of HME and HGA, which were made earlier on the basis of the clinical-serologic survey. In 10.0% of the tested taiga ticks were detected DNA of two and more agents in various combinations i.e. E. muris and Rickettsia spp, A. phagocytophilum and Rickettsia spp., and E. muris, A. phagocytophilum and Rickettsia spp. DNA of a R. helvetica was detected in I. persulcatus tick and blood tick-bitten patient with febrile episodes. Probably that R. helvetica can be etiological agent in some part of cases with the serologically unconfirmed diagnoses of acute feverish diseases developing after tick bite.

[Genetic heterogeneity of Borrelia afzelii in the natural focus of the Middle Urals].

As shown by sequencing the spacer rrf (5S)--rrl (23S) in 72 isolates of B. afzelii (one of the causative agents of Ixodes tick borne Borrelia infections) and the chromosomal gene coding protein P66 in 22 isolates, that in the natural focus located in the Middle Urals two different genetic subgroups (VS461 and NT28) of this genospecies simultaneously circulate. These subgroups are represented by 5 gene variants (rrf) 5S--(rrl) 23S and 5 allelic variants in gene p66. The latter, similarly to spacer gene variants, are not linked with a definite host and occur in different rrf--rrl variants of the infective agent. At the same time the definite species of vectors and carriers may be the host of several different B. afzelii variants, both in the spacer and in the gene coding protein P66, which maintains the genetic heterogeneity of B. afzelii population in the natural focus.

[Molecular diagnosis and frequencies of primary hypolactasia in populations of RUSSIA and neighboring countries].

The digestion of the milk sugar (lactose) is observed every normal child but not in every adult. The decreased lactase synthesis in some adults results in problems with digestion of the whole milk (primary hypolactasy). An association of lactase activity in adults with carrying of the allele T within the polymorphism C/T-13910 located upstream of the lactase gene and 100% association of hypolactasy with the genotype C/C has recently been shown for a Finnish sample. In the present work we determined the LCT* C/T_13910 genotypes and allele frequencies in populations from Russia. The genotype C/C frequencies varied from 36.6% for Russians to 88.2% for Chukchi and were close to the published medical and epidemiological data on hypolactasy frequencies in respective populations. Genotyping was performed by three different methods to identify the optimal one. Our results have shown that the studied locus is the key determinant for the primary hypolactasy development in various human populations. Consequently, the DNA diagnostics of the C/C genotype carrying is a promising predictive test to detect the primary hypolactasy long before its clinical development. Practical application of this type of diagnostics would be a step towards the individual-oriented medicine.

[Genetic variants of Borrelia afzelii, a pathogen of the ixodes tick borrelioses].

Borrelia afzelii is one of two most important pathogens of the ixodes tick borrelioses (ITB) in Russia and neighboring countries. This pathogen circulates in various ecosystems and has a wide range of reservoir hosts and transmitters. The results of studies of genetic heterogeneity of the spirochaetae B. afzelii are considered. A total of 139 primary isolates were studied. The isolates were isolated from three species of Ixodes ticks at different stages of development and obtained from the laboratory of infection transmitters, Gamaleya Scientific Research Institute of Epidemiology and Microbiology, Russran Academy of Medical Sciences, Moscow. The transmitters and reservoir hosts of borrelias were caught in natural foci of Russia (from Kaliningrad region irk the west to south Sakhalin in the east), Czechia, Lithuania, Estonia, Ukraine, and Moldavia. Analysis of genotype sequence similarity obtained by sequencing of the rrf(SS)-rrl(23S) spacer demonstrated that the B. afzelii genospecies incorporated no less than 10 genetic variants of spirochaetae, most variants being geographically widespread.

[Genetic characterization of pathogenic Borrelia, group A14S, isolated in Ukraine].

Borrelia Ir-5215, isolated from ticks Ixodes ricinus in Ukraine (the Crimean autonomous region), was identified by the method of the polymorphism of the fragment length of the restriction amplicon of rRNA spacer region 5S-23S. Its Msel-restriction profile was relatively similar to that of B. afzelii. The sequencing of spacer region rrf (5S)-rrl (23S) and 16S rRNA gene, as well as the analysis of the similarity of nucleotide sequences, obtained in the course of these study, revealed the differences between Borrelia sp, lr-5215 and six European species of Borrelia burgdorferi sensu lato and a high level of similarity (more than 95.1% for 5S-23S rRNA and 99.4% for 16S rRNA gene) to three known representatives of genome group A14S (Borrelia spp. A14S, I-77 and PC-Rq17). This suggests that isolated Borrelia lr-5215 is a new representative of pathogenic B. burgdorferi sensu lato genome group A14S, which is spread, together with Central Europe, also in southern Ukraine.

[Genetic characteristics of human pathogenic Borrelia isolated from Ixodes trianguliceps Bir. and Ixodes pavlovskyi Pom].

To support the results of typing the amplicon of the spacer site 5S-23S of pRNA by restriction fragment length polymorphism (RFLP) assay, the isolates from the ticks I. trianguliceps and I. pavlovskyi underwent sequenation of rrf (5S)-rrl (23S) spacer to make a comparative analysis of derived nucleotide sequences with the DDBJ/EMBL/ GenBank databases. For this purpose, the authors used six isolates (three imago and three nymphal) from I. trianguliceps and three isolates from adult hungry I. pavlovskyi females from the Borrelia museum of the Laboratory of Infection Transmitters, N. F. Gamaleya Research Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences. In the natural foci of the Perm Region, in the tick I. trianguliceps the same genetic variants of B. afzelii VS461 and B. garinii NT29 was found to occur as in I. persulcatus and small mammals, the reservoir Borrelia hosts. Borreliae previously detected in the tick I. pavlovskyi belong to B. garinii 20047.

[Comparative evaluation of effects of p-tyrosol and Rhodiola rosea extract on bone marrow cells in vivo]. / Sravnitel'naia otsenka deistviia p-tirozola i ékstrakta podioly rozovoi na kletki kostnogo mozga in vivo.

The effects of p-tyrosol and Rhodiola rosea extract on the hemopoietic system were compared on a model of subacute lead intoxication. No significant differences between the activity of two preparations were revealed by the study of plasma membrane blebbing, apoptosis, and necrosis processes in bone marrow. At the same time, p-tyrosol exhibited a more pronounced effect upon lipid peroxidation and offered significant protection against lead intoxication.

[Indication of Borrelia burgdorferi sensu lato in the taiga tick larvae obtained from spontaneously infected females]. / Indikatsiia Borrelia burgdorferi sensu lato v lichinkakh taezhnogo kleshcha, poluchennykh ot spontanno infitsirovannykh samok.

Hungry larvae from 17 clutches by spontaneously infected Ixodes persulcatus females were examined by inoculation of the BSK II medium, by microscopy of fixed smears, and by polymerase chain reaction (PCR) with the primers specific toward the conserved sequences of Borrelia burgdorferi 16S ribosomal RNA gene. A study of 781 larvae individually or as pools could not reveal Borrelia DNA by PCR. Inoculations of the BSK II medium with 600 larvae yielded no positive results either; immobile spiral forms of Bacillus spp. were detected in 16 (26.7 +/- 5.7%) cultures of the larvae obtained from 7 females. Microscopy of 1416 fixed smears showed typical Borrelia in 7 (0.5 +/- 0.4%) specimens of larvae resulting from the same hatch; spirochaete-like cells were present in 13 (0.9 +/- 0.5%) cases. The single female from those of the 17 hatches transmitted Borrelia through eggs to filial-generation larvae. The infection rate of the larvae emerged from the eggs of this hatch was 7% and their individual infection rates were 0.4-0.8 Borrelia per 100 fields of vision. The findings suggest that there may a transovarian transmission of Borrelia in I. persulcatus ticks, but its likelihood is very little.

[Oxidative stress and zinc chloride-induced gamete impaired morphology]. / Okislitel'nyi stress i narushenie formologii gamet, indutsiruemye khloridom tsinka.

An in vivo study investigated the toxic effect of zinc chloride on murine spermatozoids. Acute and subacute intoxication was found to increase the quantity of TBC active products and to induce the growth of degenerative forms of spermatozoids.

Role of S2 receptors in the stimulatory effect of serotonin on hemopoietic bone marrow stem cells.

Studies by the method of splenic exocolonies with serotonin receptor blockers methysergide and cyproheptadine showed that S2 receptors are involved in the stimulatory effect of serotonin on hemopoietic bone marrow stem cells.

Early effects of cis-dichlorodiammine platinum (II) on tumor progression and programmed death of Ehrlich ascites carcinoma cells.

We studied the effects of cis-dichlorodiammine platinum (II) on different pathways of cell death in cultured Ehrlich ascites carcinoma in vitro and on subsequent growth of transplanted tumor in vivo. One-hour incubation with the cytostatic modulated apoptosis in cell culture. However, exposure of cell culture to a minimal effective concentration of cis-platinum(II)diammine dichloride promoted the growth of transplanted tumor.