A preliminary study of parasite diversity in the anadromous Kamchatkan mikizha Parasalmo mykiss mykiss (Osteichthyes: Salmoniformes: Salmonidae)

The study of parasites associated with the anadromous mikizha from several rivers of the Western Kamchatka has revealed 44 parasite species. The majority of parasites (59%) belong to marine and estuarine-marine species. Lecithophyllum bothryophorum, Echinorhynchus leidyi and plerocercoids of the family Litobothriidae were found for the first time in anadromous fish entering into Asian continental watersheds. It has been revealed that only a few specimens of the anadromous mikizha feed in fresh waters. Most of feeding fish consume a minor amount of food, and this feeding is occasional. Young forms of intestinal parasites of the freshwater and estuarine-freshwater groups (Neoechinorhynchus spp., Crepidostomum spp., Cucullanus truttae, Eubothrium salvelinf) come into anadromous fishes mainly before their entering in fresh waters.     

Ion permeation through a voltage- sensitive gating pore in brain sodium channels having voltage sensor mutations

Voltage-gated sodium channels activate in response to depolarization, but it is unknown whether the voltage-sensing arginines in their S4 segments pivot across the lipid bilayer as voltage sensor paddles or move through the protein in a gating pore. Here we report that mutation of pairs of arginine gating charges to glutamine induces cation permeation through a gating pore in domain II of the Na(V)1.2a channel. Mutation of R850 and R853 induces a K(+)-selective inward cationic current in the resting state that is blocked by activation. Remarkably, mutation of R853 and R856 causes an outward cationic current with the opposite gating polarity. These results support a model in which the IIS4 gating charges move through a narrow constriction in a gating pore in the sodium channel protein during gating. Paired substitutions of glutamine allow cation movement through the constriction when appropriately positioned by the gating movements of the S4 segment.     

Identification and properties of complexes formed by myeloperoxidase with lipoproteins and ceruloplasmin

The first evidence of multi-component complexes formed by myeloperoxidase (MPO), ceruloplasmin (CP), and very low/low density lipoproteins (VLDL/LDL) obtained by electrophoresis, gel filtration, and photon-correlation spectroscopy (PCS) is presented in this paper. Complexes were observed when isolated MPO, CP, and VLDL/LDL were mixed and/or when MPO was added to the blood plasma. Complex LDL–MPO–CP was detected in 44 of 100 plasma samples taken from patients with atherosclerosis, and 33 of 44 samples also contained the VLDL–MPO–CP complex. MPO concentration in these patients’ plasma exceeded 800 ng/ml. Interaction of MPO with high density lipoproteins (HDL) was not revealed, as well as binding of CP to lipoproteins in the absence of MPO. Adding antibodies against apoB-100 to VLDL–MPO–CP and LDL–MPO–CP complexes results in release of lipoproteins. Using PCS the diameters of complexes under study were evaluated. By comparing concentrations of the components in complexes formed by MPO, CP, and lipoproteins their stoichiometry was assessed as 2VLDL:1MPO:2CP and 1LDL:1MPO:2CP. Lipoproteins affected the inhibition of MPO peroxidase activity by CP. The affinity of lipoproteins to MPO–CP complex was assessed using apparent dissociation constants determined as ～0.3 nM for VLDL and ～0.14 nM for LDL.     

Influence of human B-casomorphin-7 on specific binding of 3H-spiperone to the 5-HT2-receptors of rat brain frontal cortex

Using radio-receptor analysis, it has been demonstrated that human beta-casomorphin-7 (Tyr-Pro-Phe-Val-Glu-Pro-Ile) displaces 3H-spiperone from 5-HT2-receptors of rat brain frontal cortex. IC50 of human beta-casomorphin-7 was 8 microM. These data suggest that one of the mechanisms of neurotropic action of beta-casomorphin-7 is might be associated with its influence on the serotoninergic system.     

Dracunculoid nematodes (Spirurida: Dracunculoidea) of fishes from the Volga River delta

Faunistic and some morphological data, as well as nomenclature notes on dracunculoid nematodes parasitising fishes in the Volga River delta, are presented. The author replaced a preoccupied generic name Molnaria Moravec, 1968 (Skrjabillanidae) by the new name Kalmanmolnaria nom. nov. The validity of Philometroides lusii (Vismanis, 1962) comb. nov. as a senior objective synonym of Philometroides lusiana (Vismanis, 1967) Ivaschkin et al., 1971 is restored.     

Quantitative estimation of the dynamics of adventive flora (by the example of the Tula region)

The rate of enrichment of the Tula region flora with adventive species was quantitatively estimated taking into account the changes of their degree of naturalization during the last 200 years. Numerical score of degree of the naturalization for each species was used to compile the initial database: "0", species absent from the territory; "1", ephemerophyte; "2", colonophyte; "3", epecophyte; "4", argiophyte; "?", lack of data. Non-interpolated integral index of the dynamics of adventive flora NI(t) was calculated from this database. This index displays the sum of the degrees of naturalization of all the adventive species in the flora in some particular year. The interpolation of the initial database, aimed at minimizing the influence of random factors (e.g., gaps in observations or different activity of the researchers in different years), was performed by substituting the "?" symbol by a series of intermediate values based on studies of the data for adjacent territories. Interpolated integral indices I(t) were calculated from the interpolated database. These indices were then leveled out with Morlet wavelets, in order to distinguish random spikes (lasting less than 50 years) from the analyzed signal, and thus approximate the index dynamics to the objective trend that represents the dynamics of the flora and not the rate of activity of the researchers. The dynamics of the adventive flora of the Tula region revealed with this method shows the following facts: 1) average rate of the enrichment of the adventive flora with strange species has been constant for these 200 years and amounted to 15 species per decade; 2) average rate of naturalization was relatively low and constant, amounting to 5 species per decade; 3) fluctuations of the composition and naturalization degree of the Tula region adventive flora species were not shown to be dependant directly on the changes in the territory's economic development during the last two centuries; 4) no periodicity was recorded in the advent of new species, and the fluctuations of the number of adventive species can be attributed to the fluctuations of research intensity.     

Evenly tritium-labeled peptides and their in vivo and in vitro biodegradation

Biologically active peptides evenly labeled with tritium were used for studying the in vitro and in vivo biodegradation of the peptides. Tritium-labeled peptides with a specific radioactivity of 50-150 Ci/mmol were obtained by high temperature solid phase catalytic isotope exchange (HSCIE) with spillover tritium. The distribution of the isotope label among all amino acid residues of these peptides allows the simultaneous determination of practically all possible products of their enzymatic hydrolysis. The developed analytical method includes extraction of tritium-labeled peptides from organism tissues and chromatographic isolation of individual labeled peptides from the mixture of degradation products. The concentrations of a peptide under study and the products of its biodegradation were calculated from the results of liquid scintillation counting. This approach was used for studying the pathways of biodegradation of the heptapeptide TKPRPGP (Selank) and the tripeptide PGP in blood plasma. The pharmacokinetics of Selank, an anxiolytic peptide, was also studied in brain tissues using the intranasal in vivo administration of this peptide. The concentrations of labeled peptides were determined, and the pentapeptide TKPRP, tripeptide TKP, and dipeptides RP and GP were shown to be the major products of Selank biodegradation. The study of the biodegradation of the heptapeptide MEHFPGP (Semax) in the presence of nerve cells showed that the major products of its biodegradation are the pentapeptide HFPGP and tripeptide PGP. The enkephalinase activity of blood plasma was studied with the use of evenly tritium-labeled [Leu]enkephalin. A high inhibitory effect of Semax on blood plasma enkephalinases was shown to arise from its action on aminopeptidases. The method, based on the use of evenly tritium-labeled peptides, allows the determination of peptide concentrations and the activity of enzymes involved in their degradation on a tg scale of biological samples both in vitro and in vivo.     

Expression of an expanded polyglutamine domain in yeast causes death with apoptotic markers

Huntington's disease is caused by specific mutations in huntingtin protein. Expansion of a polyglutamine (polyQ) repeat of huntingtin leads to protein aggregation in neurons followed by cell death with apoptotic markers. The connection between the aggregation and the degeneration of neurons is poorly understood. Here, we show that the physiological consequences of expanded polyQ domain expression in yeast are similar to those in neurons. In particular, expression of expanded polyQ in yeast causes apoptotic changes in mitochondria, caspase activation, nuclear DNA fragmentation and death. Similar to neurons, at the late stages of expression the expanded polyQ accumulates in the nuclei and seems to affect the cell cycle of yeast. Interestingly, nuclear localization of the aggregates is dependent on functional caspase Yca1. We speculate that the aggregates in the nuclei disturb the cell cycle and thus contribute to the development of the cell death process in both systems. Our data show that expression of the polyQ construct in yeast can be used to model patho-physiological effects of polyQ expansion in neurons.     

Evenly tritium labeled peptides in study of peptide in vivo and in vitro biodegradation

Biologically active peptides evenly labeled with tritium were used for studying the in vitro and in vivo biodegradation of the peptides. Tritium-labeled peptides with a specific radioactivity of 50–150 Ci/mmol were obtained by high temperature solid phase catalytic isotope exchange (HSCIE) with spillover tritium. The distribution of the isotope label among all amino acid residues of these peptides allows the simultaneous determination of practically all possible products of their enzymatic hydrolysis. The developed analytical method includes extraction of tritium-labeled peptides from organism tissues and chromatographic isolation of individual labeled peptides from the mixture of degradation products. The concentrations of a peptide under study and the products of its biodegradation were calculated from the results of liquid scintillation counting. This approach was used for studying the pathways of biodegradation of the heptapeptide TKPRPGP (Selank) and the tripeptide PGP in blood plasma. The pharmacokinetics of Selank, an anxiolytic peptide, was also studied in brain tissues using the intranasal in vivo administration of this peptide. The concentrations of labeled peptides were determined, and the pentapeptide TKPRP, tripeptide TKP, and dipeptides RP and GP were shown to be the major products of Selank biodegradation. The study of the biodegradation of the heptapeptide MEHFPGP (Semax) in the presence of nerve cells showed that the major products of its biodegradation are the pentapeptide HFPGP and tripeptide PGP. The enkephalinase activity of blood plasma was studied with the use of evenly tritium labeled [Leu]enkephalin. A high inhibitory effect of Semax on blood plasma enkephalinases was shown to arise from its action on aminopeptidases. The method, based on the use of evenly tritium-labeled peptides, allows the determination of peptide concentrations and the activity of enzymes involved in their degradation on a μg scale of biological samples both in vitro and in vivo.