First Data on the Contact Zone and Hybridization between the Cryptic Species of Shrews Sorex araneus and S. satunini (Eulipotyphla, Mammalia).

For the first time, based on sequence variation of microsatellite loci and the mtDNA cytb gene fragment, population genetic structure of the common shrew and Caucasian shrew in their contact zone was investigated. It was demonstrated that, although there was no complete reproductive isolation between the species under consideration, the gene flow was considerably limited. These data testify to the established reliable reproductive barriers between the common shrew and Caucasian shrew.

Effect of chaperonin encoded by gene 146 on thermal aggregation of lytic proteins of bacteriophage EL Pseudomonas aeruginosa.

Investigation of the chaperonin encoded by gene 146 of bacteriophage EL Pseudomonas aeruginosa that we characterized earlier has been continued. To reveal the mechanism of its functioning, new recombinant substrate proteins, fragments of gene product (gp) 183 containing the lysozyme domain were prepared. Their interaction with gp146 was studied. The influence of the phage chaperonin on the thermal aggregation of one of these gp183 fragments and endolysin (gp188) was investigated in both the presence and the absence of ATP by dynamic light scattering. In the absence of ATP, the phage chaperonin forms stable complexes with substrate proteins, thereby protecting them against thermal aggregation. Experimental data obtained for different substrate proteins are analyzed.

[Genetic Structure of the Common Shrew Sorex araneus L. 1758 (Mammalia, Lipotyphla) in Continuous and Fragmented Areas].

In this work the genetic variability of the common shrew populations Sorex araneus L. in Eastern Europe was studied via sequencing of the mitochondrial gene cyt b. A total of 82 sequences of the mitochondrial gene cyt b with a length of 953 basepairs were analyzed, including five chromosome races in a continuous area of the species in forest zone and two races in fragmented area in the steppe zone. Phylogeographic subdivision of the common shrew was not expressed, and there was no significant correlation between genetic and geographic distances in continuous areas. We did not acquire convincing evidence of the influence of narrow hybrid zones between chromosome races on the flow of neutral alleles. A significant p-distance (0.69 ± 0.27%) of geographically close populations of the chromosome race Neroosa indicates the formation of the karyotype of this race in the Pliocene or Pleistocene. In our work, the phylogeographic structure was determined more by species area fragmentation than by its karyotypic features.

Complexes between anionic liposomes and spherical polycationic brushes. An assembly of assemblies.

This paper has at its objective the assembling of liposomal assemblies onto nanoparticles. In this manner, one generates nanoparticles with a high loading capacity. Thus, spherical spherical polycationic "brushes" (SPBs) were synthesized by graft polymerizing a cationic monomer, (trimethylammonium)ethylmethacrylate chloride, onto the surface of monodisperse polystyrene particles, ca. 100 nm in diameter. These particles were complexed with small unilamellar anionic liposomes, 40-60 nm in diameter, composed of egg lecithin (EL) and anionic phosphatidylserine (PS(1-)) in PS(1-)/EL ratios from 0.10 to 0.54, a key parameter designated as ν. These complexes were then characterized according to electrophoretic mobility, dynamic light scattering, conductivity, fluorescence, and cryogenic transmission electron microscopy, with the following main conclusions: (a) All added liposomes are totally associated with SPBs up to a certain saturation concentration (specific for each ν value). (b) The number of liposomes per SPB particle varies from 40 (ν = 0.1) to 14 (ν = 0.5). (c) At sufficiently high liposome concentrations, the SPBs experience an overall change from positive to negative charge. (d) SPB complexes tend to aggregate when their initial positive charge has been precisely neutralized by the anionic liposomes. Aggregation is impeded by either positive charge at lower lipid concentrations, or negative charge at higher lipid concentrations. (e) The liposomes remain intact (i.e., do not leak) when associated with SPBs, at ν ≤ 0.5. (f) Complete SPB/liposome dissociation occurs at external [NaCl] = 0.3 M for ν = 0.1 and at 0.6 M for ν = 0.5. Liposomes with ν = 0.54 do not dissociate from the SPBs even in NaCl solutions up to 1.0 M. (g) Complexation of the PS(1-)/EL liposomes to the SPBs induces flip-flop of PS(1-) from the inner leaflet to the outer leaflet. (h) The differences in the ability of PS(1-) (a cylindrical lipid) and CL(2-) (a conical lipid) to create membranes defects are attributed to geometric factors.

[Craniometric differences between karyotypic races of the common shrew Sorex araneus (Mammalia) as a result of limited hybridization].

The contact points of four karyotypic races (St. Petersburg, Moscow, Seliger and West Dvina) of the common shrew Sorex araneus L. were studied at the Valdai Hills (European Russia) in an area unimpeded by geographic barriers. The populations of the races are separated by narrow hybrid zones that represent the most complex heterozygous hybrid karyotypes. At these points of contact, the morphometric differentiation of karyotype races was examined in 12 cranial measurements in 190 shrews of a known karyotype. A. comparison of the mean values in studied samples of immature shrews revealed statistically significant differences and the correlation of some measurements in order to describe the level of musculus temporalis. It has been proposed that morphometric differences in the karyotypic races were preserved and accumulated because of a 50% reduction of the frequencies of hybrids. The deviation from the Hardy-Weinberg ration in the frequencies of the genotype and haploid sets of chromosomes in the hybrid zones can be attributed to a number of fatalities of hybrid embryos or the nonrandom mating of karyotypic races. The ethological isolation might arise in the evolution of some karyotypic races from the reduced fitness of the hybrids.

[Narrow hybrid zone between Moscow and Western Dvina chromosomal races and specific features of population isolation in common shrew Sorex araneus (mammalia)].

The contact zone between Moscow and Western Dvina chromosomal races of common shrew Sorex araneus L. at the south of the Valdai Hights was traced over a distance of 20 km. Within this, close to parapatric, contact zone of chromosomal races the width of sympatry zone was about 500 m (the narrowest among currently known hybrid zones), and the proportion of hybrids was 24.3%. It was shown that in bimodal hybrid zones between chromosomal races of common shrew the width of sympatry zones varied from 0.5 to 13 km. This width does not correlate with the cytogenetic features of the hybrids, and seems to be determined by competitive relations between the races. The hybrid proportion is determined by the type of hybrid heterozygosity, and decreased in the race sympatry zone from 33-40 to 21.5-25.2%. The decrease of the hybrid proportion can be associated with the abnormal fertility of either the first generation, or the backcross hybrids.

[Phylogenetic relationships of Caucasian shrew Sorex satunini Ogn. (mammalia) in the superspecies Sorex araneus inferred from the data of karyological analysis and the mtDNA cyt b gene sequencing].

Using the data of karyological analysis, the phylogenetic relationships of Caucasian shrew Sorex satunini and the cryptic species of superspecies Sorer araneus were examined. In the population of Sorex satunini from the plain of North Ciscaucasia two deeply radiated cytochrome b genes (A and B) were identified. Genetic distance between haplotype A and B groups constituted 0.0675 +/- 0.008, which is higher than any distance in superspecies S. araneus. Possible introgression of type B haplotypes from the populations of the evolutionary lineage S. subaraneus--S. araneus in Pleistocene and the time of the appearance of the chromosomal polymorphism of S. araneus is discussed. Our results show that the use of only one mitochondrial marker can lead to false conclusions on taxonomic diversity

[The microsatellite polymorphism and gene flow in the contact zone of four common shrew (Sorex araneus L., Mammalia) chromosome races].

The variation of microsatellite loci in 130 individuals of four common shrew chromosome races (Moscow, Western Dvina, Seliger, and St. Petersburg) contacting on the Valdai Hills was studied. A low level of genetic differences between the chromosome races, which differ at three-five fixed diagnostic metacentric chromosomes, was found. The genetic differentiation within the races is more considerable as compared with that between the races. A high deficiency in heterozygotes was recorded; presumably, this is connected with regular variation in the population sizes. It is assumed that the fixation of centric chromosome fusions was supported by selection (drive) in the evolution of the common shrew against the background of a neutral evolution of the microsatellite loci.

[Correlation between morphological and genetic differentiation in Cuban hutia (Rodentia, Capromyinae): species fragmentation on the area and the problem of excessive taxonomic status problem].

The work attempts to explain the previously discovered phenomenon of excessive taxonomic status of the species and subspecies of Cuban hutias relative to their protein genetic divergence. The reasons underlying the fragmentary distribution of the species belonging to the family Capromyidae over the Antilles and Bahamas is also considered. Samples of four geographically distant hutia (genera Capromys and Mysateles) populations inhabiting different biotypes were assessed according to 32 allozyme loci. It has been demonstrated that the interpopulation and subspecies differentiation in the gene frequencies in these genera is very low. As for the pattern of polymorphism (Shannon?s measure), the populations and subspecies within these genera are well differentiated and adequate to their morphological differentiation. This suggests that the genetic divergence pattern of hutias in post-Pleistocene was predominantly determined by selection on the background of a relatively rapid formation of multiple morphotypes. It is assumed that the evolution within the family Capromyidae in the earlier and the later periods was associated with the geomorphological and climatic events in the history of the Earth. This standpoint provides for explaining the phenomena of both the excessive taxonomic status of the Cuban Capromyinae and the fragmentary distribution of these species over the Antilles and Bahamas, amazing at a first glance.

[Fixation of metacentric chromosomes in populations of the common shrew Sorex araneus L. from populations of Eastern Europe].

In this review, we discuss the processes of fixation of Robertsonian chromosome fusions in populations of the common shrew Sorex araneus L. Various Robertsonian fusions, accumulating in populations, create an illusion of large chromosomal rearrangements, reciprocal translocations of complete chromosome arms. The use of these rearrangements for phylogenetic reconstructions results in false conclusions. Robertsonian fusions accumulate in populations at such stages of the species evolution, when large open or subdivided populations prevail (populations of warm periods of Pleistocene and many present-day populations) and are fixed in small isolated populations and glacial refugia. The formation of monomorphic chromosome races requires a long time, several glaciation epochs during the whole Pleistocene.

[On the taxonomic rank of ciscaucasicus and its relationships with the pygmy wood mouse Sylvaemus uralensis inferred from the mtDNA cytochrome b gene sequence].

To specify the taxonomic rank of form ciscaucasoides (independent species Sylvaemus ciscaucasoides, or intraspecific form of pygmy wood mouse, S. uralensis), a 402-bp the mtDNA cytochrome b gene fragment (402 bp) was examined in S. ciscaucasoides individuals from six geographic localities of the Caucasus and Ciscaucasus, (Krasnodar krai and Adygeya Republic) and 17 S. uralensis individuals from seven localities of the Russian Plai (Saratov oblast, Smolensk oblast, Voronezh oblast, Tula oblast, Moscow oblast, and Tver' oblast). For comparison, the cytochrome b gene was partly sequenced in the samples of yellow necked, S. flavicollis (n = 2, Samara oblast), and Caucasian, S. ponticus (n = 6, Krasnodar krai), wood mice. One Mus musculus specimen from Western Europe, whose nucleotide sequences were deposed in the GenBank, was used as intergeneric outgroup. Phylogenetic trees for the forms examined were constructed based on the mtDNA sequence variation and using the neighbor joining and maximum parsimony methods. The network of the cytochrome b haplotypes was also constructed. The level of genetic divergence was evaluated using Kimura's two-parameter algorithm. Based on the data on the sequence variation in a 402-bp mtDNA cytochrome b gene fragment, the hypothesis on the species status of the ciscaucasicus form was. The mean intergroup distances (d) between the geographic groups of S. uralensis varied from 0.0036 to 0.0152. At the same time, the distances between the pygmy wood mice and the group of S.flavicollis-S. ponticus varies in the range from 0.0860 to 0.0935, and the level of intergeneric genetic differentiation (Sylvaemus-Mus) is higher than the latter index (d = 0.142). Ciscaucasoides should be considered as geographic substitution form of S. uralensis. Furthermore, the Caucasian populations of S. uralensis (= ciscaucasoides) were characterized by a threefold lower value of intergroup genetic divergence (d = 0.0062) than the East European populations (d= 0.0179). This finding pointed to some isolation of Caucasian populations of pygmy wood mouse and depletion of their gene pool. However other molecular genetic data (similarity of nucleotide composition and consistence of the levels of intra- and intergroup distances) suggest the absence of geographic subdivision between Caucasian and East European populations ofS. uralensis relative to the molecular marker examined.

[Potentialities of multispiral computed tomography in the diagnostics of pathological alterations of the colon].

The work describes an experience with using different methods of multispiral computed tomography in the diagnosis and differential diagnostics of pathological changes in the colon of patients of colonoproctological profile. Methodical variations of the computed tomography investigations are described in patients with localization of tumors in different parts of the intestine.

[Using AELTIS-synchro-02 device in the therapy of chronic bacterial prostatitis].

The treatment of chronic bacterial prostatitis combined antibacterial drugs and physiotherapy (low-energy laser radiation, electrostimulation of the prostate). Treatment of chronic bacterial prostatitis with medication and complex two-channel bio-synchronized electrolaser therapy with application of the unit AELTIS-synchro-02 raises efficacy of treatment with chronic bacterial prostatitis due to combined effect of antibacterial drugs and bacteriostatic and immunomodulating actions of the physical factors applied. These normalize microcirculation in the region of the prostatic gland, improve a draining function of the prostatic ducts, allows achievement of good results in 88.2% patients.

Combination of the electrogenic ionophores, valinomycin and CCCP, can lead to non-electrogenic K+/H+ exchange on bilayer lipid membranes.

The method of pH shift measuring by means of a pH microelectrode was applied to measure hydrogen ion fluxes across a planar bilayer lipid membrane (BLM) in the presence of the potassium ion ionophore, valinomycin, and a protonophore, carbonylcyanide m-chlorophenylhydrazone (CCCP), under conditions of the voltage clamp. The voltage dependence of the flux was determined to be in the range of +/- 150 mV under the conditions of both symmetrical KCl as well as a KCl gradient across the BLM. Surprisingly, at a clamped zero voltage on BLM a significant hydrogen ion flux was observed in the presence of a KCL gradient and both valinomycin and CCCP. This finding was interpreted as a result of induction of non-electrogenic K+/H+ exchange in the presence of valinomycin and CCCP, presumably through the formation of electrically neutral complexes of these two ionophores and K+ (H+) ions: valinomycin-K(+)-CCCP- and/or possibly valinomycin-CCCP(-)-H+.

Thermally induced chain exchange of smooth muscle tropomyosin dimers studied by differential scanning calorimetry.

The thermal unfolding of duck gizzard tropomyosin dimers, alphabeta, alphaalpha, and betabeta, and of a 1:1 mixture of alphaalpha and betabeta homodimers was studied by differential scanning calorimetry (DSC). Both alphaalpha and betabeta homodimers demonstrated a broad thermal transition with maxima at 37.4 degrees C and 44.6 degrees C, respectively. However, a sharp cooperative thermal transition at 41.5 degrees C characteristic for alphabeta heterodimer appeared on the thermogram of the mixture of homodimers. The appearance of this transition was prevented by disulfide cross-linking of polypeptide chains in the homodimers. Thus, DSC studies clearly demonstrate formation of tropomyosin heterodimers during heating of the mixture of homodimers and in agreement with earlier published reports indicate thermally induced chain exchange between tropomyosin dimers.

A comparative differential scanning calorimetric study of tobacco mosaic virus and of its coat protein ts mutant.

The differential scanning calorimetry (DSC) 'melting curves' for virions and coat proteins (CP) of wild-type tobacco mosaic virus (strain U1) and for its CP ts mutant ts21-66 were measured. Strain U1 and ts21-66 mutant (two amino acid substitutions in CP: 121 --> T and D66 --> G) differ in the type of symptoms they induce on some host plants. It was observed that CP subunits of both U1 and ts21-66 at pH 8.0, in the form of small (3-4S) aggregates, possess much lower thermal stability than in the virions. Assembly into the virus particles resulted in a DSC melting temperature increase from 41 to 72 degrees C for U1 and from 38 to 72 degrees C for ts21-66 CP. In the RNA-free helical virus-like protein assemblies U1 and ts21-66 CP subunits had a thermal stability intermediate between those in 3-4S aggregates and in the virions. ts21-66 helical protein displayed a somewhat lower thermal stability than U1.

Changes in the thermal unfolding of p-phenylenedimaleimide-modified myosin subfragment 1 induced by its 'weak' binding to F-actin.

Differential scanning calorimetry (DSC) was used to analyze the thermal unfolding of myosin subfragment 1 (S1) with the SH1 (Cys-707) and SH2 (Cys-697) groups cross-linked by N,N'-p-phenylenedimaleimide (pPDM-S1). It has been shown that F-actin affects the thermal unfolding of pPDM-S1 only at very low ionic strength, when some part of pPDM-S1 binds weakly to F-actin, but not at higher ionic strength (200 mM KCl). The weak binding of pPDM-S1 to F-actin shifted the thermal transition of pPDM-S1 by about 5 degrees C to a higher temperature. This actin-induced increase in thermal stability of pPDM-S1 was similar to that observed with 'strong' binding of unmodified S1 to F-actin. Our results show that actin-induced structural changes revealed by DSC in the myosin head occur not only upon strong binding but also on weak binding of the head to F-actin, thus suggesting that these changes may occur before the power-stroke and play an important role in the motor function of the head.

Shape and spreading of platelets from the blood of patients with acute myocardial infarction.

The shape of platelets in circulating blood and alterations of this shape after the contact of platelets with glass surfaces, as well as spontaneous platelet aggregation, were examined in patients with acute myocardial infarction (AMI) and in controls. The shape of circulating platelets were studied by scanning electron microscopy and the spreading--by differential interference contrast microscopy. Most platelets circulating in the blood of the patients with AMI had non-activated discoid shape; the percentage of these non-activated platelets was similar to that in the control group. However, these platelets of many AMI patients acquired an increased ability to undergo morphological changes after contact with glass. The percentage of spread platelets in controls was never higher than 50%. In contrast, this percentage was higher than 50% in 73% of patients with AMI. Increased spontaneous platelet aggregation was determined only in 30% of AMI patients.