Neurotrophic control of the transmembrane Cl− pump in mammalian muscle fibers

Resting membrane potential of both innervated and denervated rat diaphragm muscle fibers was investigated when the concentration of potential-producing ions was changed in the external fluid and following treatment with furosemide. It was found that equilibrium potential ( ) equalled resting potential level in innervated muscle for Cl^–, but shifts to more positive values compared with resting membrane potential following section of the nerve. Furosemide retards development of post-denervation depolarization of the muscle membrane. It is deduced that trophic influences originating from the motor nerve activates the furosemide-sensitive Cl^– influx system, leading to raised intracellular concentration of Cl^–, a shift in (E_Cl) and depolarization of the muscle membrane.S. V. Kurashov Medical Institute, Minsitry of Health of the RSFSR, Kazan'. Translated from Neirofiziologiya, Vol. 19, No. 6, pp. 766–771, November–December, 1987.     

Study of the antigenic structure of human immunodeficiency virus using synthetic peptides]

In a search for synthetic peptide antigens fit to detect anti-HIV antibodies, a set of algorithms were used to predict the probable antigenic determinants of gag, pol, env and nef proteins of HIV-1 and HIV-2. Over forty peptides were synthesized by the solid-phase method. The reactivity of the peptide antigens was evaluated in ELISA on panels of HIV-1/2-positive sera. Application of the synthetic peptides for the early HIV diagnostics was examined.     

Study of Secondary Specificity of Enteropeptidase in Comparison with Trypsin

A comparative study of secondary specificities of enteropeptidase and trypsin was performed using peptide substrates with general formula A-(Asp/Glu) _n -Lys(Arg)--B, where n = 1-4. This was the first study to demonstrate that, similar to other serine proteases, enteropeptidase has an extended secondary binding site interacting with 6-7 amino acid residues surrounding the peptide bond to be hydrolyzed. However, in the case of typical enteropeptidase substrates containing four negatively charged Asp/Glu residues at positions P2-P5, electrostatic interaction between these residues and the secondary site Lys99 of the enteropeptidase light chain is the main factor that determines hydrolysis efficiency. The secondary specificity of enteropeptidase differs from the secondary specificity of trypsin. The chromophoric synthetic enteropeptidase substrate G₅DK-F(NO₂)G (k _cat/K _m = 2380 mM^–1·min^–1) is more efficient than the fusion protein PrAD₄K-P26 (k _cat/K _m = 1260 mM^–1·min^–1).     

The distribution of bacterioplankton and mesozooplankton biomass in the White and Barents Seas coastal water

The total population density and the biomass of bacterioplankton, mesozooplankton, and phosphate-accumulating bacteria (PAB) were estimated during the 2000-2001 summer-autumn seasons in the coastal waters of the White and Barents Seas, which are subjects to the action of tidal and sea currents, the inflow of riverine waters, and anthropogenic impact. In the shallow estuarine waters with salinities of 6.5-32@1000 near the Chernaya, Pesha, and Pechora River mouths, the population of PAB fluctuated from 0.1 to 9.1 million cells/ml (0-36% of the total bacterial population). In pelagic seawaters, which are low in phosphates (12-50 micrograms/l) and are characterized by an increased iron/phosphorus ratio (2.0-3.6), bacterioplankton amounted to 0.1-1.6 million cells/ml and was mainly represented by small organisms with a volume of 0.08-0.15 micron 3, commonly lacking intracellular polyphosphates. In the pelagic zone of the Barents Sea, the biomass of mesozooplankton (Bz) was comparable with that of bacterioplankton (Bb = 39-175 mg/m3), the Bb/Bz ratio being 1.4-4.6. Off the Varandeiskii, Pechora, and Kolguyev oil terminals, Bb increased to 155-300 mg/m3 and the Bb/Bz ratio rose to 1.4 to 50.3 (with an average value of 20.9), presumably due to the severe anthropogenic impact on these waters. In this case, the dense population of bacterioplankton (0.9-7.6 million cells/ml) was mainly represented by large cells (0.12-0.76 micron 3 in volume), most of which (3-43% of the total bacterioplankton population) contained polyphosphates. The chemical composition of these waters was characterized by an elevated content of the total phosphorus (65-128 micrograms/l) and by a low iron/phosphorus ratio (0.9-1.2).     

Eif3ba regulates cranial neural crest development by modulating p53 in zebrafish

Congenital diseases caused by abnormal development of the cranial neural crest usually present craniofacial malformations and heart defects while the precise mechanism is not fully understood. Here, we show that the zebrafish eif3ba mutant caused by pseudo-typed retrovirus insertion exhibited a similar phenotype due to the hypogenesis of cranial neural crest cells (NCCs). The derivatives of cranial NCCs, including the NCC-derived cell population of pharyngeal arches, craniofacial cartilage, pigment cells and the myocardium derived from cardiac NCCs, were affected in this mutant. The expression of several neural crest marker genes, including crestin, dlx2a and nrp2b, was specifically reduced in the cranial regions of the eif3ba mutant. Through fluorescence-tracing of the cranial NCC migration marker nrp2b, we observed reduced intensity of NCC-derived cells in the heart. In addition, p53 was markedly up-regulated in the eif3ba mutant embryos, which correlated with pronounced apoptosis in the cranial area as shown by TUNEL staining. These findings suggest a novel function of eif3ba during embryonic development and a novel level of regulation in the process of cranial NCC development, in addition to providing a potential animal model to mimic congenital diseases due to cranial NCC defects. Furthermore, we report the identification of a novel transgenic fish line Et(gata2a:EGFP)^pku418 to trace the migration of cranial NCCs (including cardiac NCCs); this may serve as an invaluable tool for investigating the development and dynamics of cranial NCCs during zebrafish embryogenesis.     

Cyanide detoxification by microbial consortia of natural-industrial complexes of gold heap leaching

Microorganisms that have adapted not only to high concentrations of pollutants but also to environmental conditions develop in autochthonous microbial communities of natural-industrial complexes of gold heap leaching. The biotechnological potential and diversity of autochthonous microbial communities involved in cyanide detoxification was evaluated by the example of a deposit situated in the Sakha (Yakutia) Republic. Under the zoning conditions of the ore heap, the biological component had a greater impact on cyanide destruction than chemical transformation. Metabolically active representatives of a microbial consortium are capable of surviving developed under these conditions. Phylotypes of the genus Serratia and family Alcaligenaceae that are capable of cyanide destruction and are potentially promising for the detoxification of wastes of gold heap leaching were revealed.     

Intracellular Accumulation of the Monomeric Precursors of Polyphosphates and Polyhydroxyalkanoates in Acinetobacter calcoaceticusand Escherichia coliCells

The formation of polyhydroxyalkanoates granules in anaerobically grown Escherichia coliM-17 cells was found to be preceded by the intracellular accumulation of carbonic acids (predominantly, acetic acid), amounting to 9% of the cytosol. The intracellular concentration of acidic metabolites increased after the lyophilization of the bacterial biomass and decreased after its long-term storage (3.5–13.5 years). The decrease in the concentration of acidic metabolites is likely due to the dehydration of dimeric carbonic acids in the viscoelastic cytosol of resting bacterial cells. The hydrophobic obligately aerobic cells of Acinetobacter calcoaceticusIEGM 549 are able to utilize a wide range of growth substrates (from acetate and citrate to hydrophobic hydrocarbons), which is considerably wider than the range of the growth substrates of E. coli(predominantly, carbohydrates). The minimal essential and optimal concentrations of orthophosphates in the growth medium of A. calcoaceticuswere found to be tens of times lower than in the case of E. coli.The intracellular content of orthophosphates in A. calcoaceticuscells reached 35–77% of the total phosphorus content (P_total), providing for the intense synthesis of polyphosphates. The P_totalof the A. calcoaceticuscells grown in media with different proportions between the concentrations of acetate and phosphorus varied from 0.7 to 3.3%, averaging 2%. This value of P_totalis about two times higher than that observed for fermenting E. colicells. Lowering the cultivation temperature of A. calcoaceticusfrom 37–32 to 4°C augmented the accumulation of orthophosphates in the cytoplasm, presumably owing to a decreased requirement of growth processes for orthophosphate. In this case, if the concentration of phosphates in the cultivation medium was low, they were completely depleted.     

Intracellular accumulation of monomer precursors of polyphosphates and polyhydroxyalkanoates from Acinetobacter calcoaceticus and Escherichia coli]

The formation of polyhydroxyalkanoates granules in anaerobically grown Escherichia coli cells was found to be preceded by the intracellular accumulation of carbonic acids (predominantly, acetic acid), amounting to 9% of the cytosol. The intracellular concentration of acidic metabolites increased after the lyophilization of the bacterial biomass and decreased after its long-term storage (3.5-13.5 years). The decrease in the concentration of acidic metabolites is likely due to the dehydration of dimeric carbonic acids in the viscoelastic cytosol of resting bacterial cells. The hydrophobic obligately aerobic cells of Acinetobacter calcoaceticus IEGM 549 are able to utilize a wide range of growth substrates (from acetate and citrate to hydrophobic hydrocarbons), which is considerably wider than the range of the growth substrates of E. coli (predominantly, carbohydrates). The minimal essential and optimal concentrations of orthophosphates in the growth medium of A. calcoaceticus were found to be tens of times lower than in the case of E. coli. The intracellular content of orthophosphates in A. calcoaceticus cells reached 35-77% of the total phosphorus content (Ptotal), providing for the intense synthesis of polyphosphates. The Ptotal of the A. calcoaceticus cells grown in media with different proportions between the concentrations of acetate and phosphorus varied from 0.7 to 3.3%, averaging 2%. This value of Ptotal is about two times higher than that observed for fermenting E. coli cells. Lowering the cultivation temperature of A. calcoaceticus from 37-32 to 4 degrees C augmented the accumulation of orthophosphates in the cytoplasm, presumably owing to a decreased requirement of growth processes for orthophosphate. In this case, if the concentration of phosphates in the cultivation medium was low, they were completely depleted.     

Antioxidant and detoxifying activities of analogues of the delta sleep inducing peptide

Sixteen peptides differing in structure from the delta sleep inducing peptide (DSIP) by one to two substitutions of amino acid residues have been synthesized to study the possibility of their application in oncology. The antioxidant properties of the peptides in vitro and their detoxifying activity in vivo have been examined on a model of toxicosis caused by cisplatin, a cytostatic drug widely used in the therapy of tumor diseases. It has been shown that almost all DSIP analogues examined exhibit a direct antioxidant activity, with the activity of the ID-6 analogue being higher than that of DSIP and comparable with that of vitamin C and β-carotene. This analogue shows the most pronounced detoxifying activity toward the action of cisplatin, which manifests itself as a decrease in the death of animals from acute toxicity to 17% compared to 50–67% in the control and the restoration of some biochemical parameters of blood, in particular, a decrease in the activity of the enzymes, aspartate and alanine aminotransferases, and the concentration of the end products of nitrogen exchange: creatinine and urea. Thus, the peptides of the DSIP family may appear promising agents for decreasing the toxic effects of cytostatics used in oncology.     

Influence of environmental conditions on the distribution of Central Asian green toads with three ploidy levels

We studied the distribution of Palearctic green toads (Bufo viridis subgroup), an anuran species group with three ploidy levels, inhabiting the Central Asian Amudarya River drainage. Various approaches (one‐way, multivariate, components variance analyses and maximum entropy modelling) were used to estimate the effect of altitude, precipitation, temperature and land vegetation covers on the distribution of toads. It is usually assumed that polyploid species occur in regions with harsher climatic conditions (higher latitudes, elevations, etc.), but for the green toads complex, we revealed a more intricate situation. The diploid species (Bufo shaartusiensis and Bufo turanensis) inhabit the arid lowlands (from 44 to 789 m a.s.l.), while tetraploid Bufo pewzowi were recorded in mountainous regions (340–3492 m a.s.l.) with usually lower temperatures and higher precipitation rates than in the region inhabited by diploid species. The triploid species Bufo baturae was found in the Pamirs (Tajikistan) at the highest altitudes (2503–3859 m a.s.l.) under the harshest climatic conditions.