Characterization of the recombinant diaminobutyric acid acetyltransferase from Methylophaga thalassica and Methylophaga alcalica

Diaminobutyric acid acetyltransferase (EctA) catalyzes the acetylation of diaminobutyric acid to gamma-N-acetyl-alpha,gamma-diaminobutyrate with acetyl coenzyme A. This is the second reaction in the ectoine biosynthetic pathway. The recombinant EctA proteins were purified from two moderately halophilic methylotrophic bacteria: Methylophaga thalassica ATCC 33146T and Methylophaga alcalica ATCC 35842T. EctA found in both methylotrophs is a homodimer with a subunit molecular mass of c. 20 kDa and had similar properties with respect to the optimum temperature for activity (30 degrees C), Km for diaminobutyrate (370 or 375 microM) and the absence of requirements for divalent metal ions. The enzyme from M. thalassica exhibited a lower pH optimum and was inhibited both by sodium carbonates and by high ionic strength but to a lesser extent by copper ions.     

Interactions of the introduced rotan Perccottus glenii Dybowski, 1877 (Osteichthyes, Odontobutidae) with aboriginal fish species: The parasitological aspect

This study was conducted in the water-filled Neverovo sandpit, located in Ruza district of Moscow province. This water body is inhabited by 6 fish species, including the introduced rotan. Twelve species of parasites were recorded in/on the rotan, and most of them circulate using aboriginal fish species. Reliable evidence of the involvement of the rotan in the circulation of a parasite species native to the water body-recipient (Ergasilus sieboldi) was obtained for the first time. Rotan does not play a significant role in the circulation of 11 other parasite species, or, at least, quantity estimations of its role were not yet undertaken.     

Variation in the timing of spawning of the black sea brown trout <Emphasis Type="Italic">Salmo trutta labrax</Emphasis> pallas under artificial and natural conditions

Analysis of the maturation and spawning times of the Black Sea brown trout bred at the fish-farming plants and inhabiting natural waterways of the Northwestern Caucasus has demonstrated a considerable variation depending on environmental conditions, first and foremost, temperature. This fact, as well as the analysis of literature data, suggests that the duration and timing of the spawning season cannot be used as self-sufficient criteria for identifying species of the genus Salmo.     

Fetal Serum in Combination with 5% Dimethyl Sulfoxide Efficiently Protects the Human Gut Microbiota during Cryopreservation in Liquid Nitrogen

Biophysics - Due to advances in medical technologies, it is an important problem to develop methods that allow safe and efficient long-term preservation of the human gut microbiota without...     

Water-soluble LYNX1 Residues Important for Interaction with Muscle-type and/or Neuronal Nicotinic Receptors

Human LYNX1, belonging to the Ly6/neurotoxin family of three-finger proteins, is membrane-tethered with a glycosylphosphatidylinositol anchor and modulates the activity of nicotinic acetylcholine receptors (nAChR). Recent preparation of LYNX1 as an individual protein in the form of water-soluble domain lacking glycosylphosphatidylinositol anchor (ws-LYNX1; Lyukmanova, E. N., Shenkarev, Z. O., Shulepko, M. A., Mineev, K. S., D''Hoedt, D., Kasheverov, I. E., Filkin, S. Y., Krivolapova, A. P., Janickova, H., Dolezal, V., Dolgikh, D. A., Arseniev, A. S., Bertrand, D., Tsetlin, V. I., and Kirpichnikov, M. P. (2011) NMR structure and action on nicotinic acetylcholine receptors of water-soluble domain of human LYNX1. J. Biol. Chem. 286, 10618–10627) revealed the attachment at the agonist-binding site in the acetylcholine-binding protein (AChBP) and muscle nAChR but outside it, in the neuronal nAChRs. Here, we obtained a series of ws-LYNX1 mutants (T35A, P36A, T37A, R38A, K40A, Y54A, Y57A, K59A) and examined by radioligand analysis or patch clamp technique their interaction with the AChBP, Torpedo californica nAChR and chimeric receptor composed of the α7 nAChR extracellular ligand-binding domain and the transmembrane domain of α1 glycine receptor (α7-GlyR). Against AChBP, there was either no change in activity (T35A, T37A), slight decrease (K40A, K59A), and even enhancement for the rest mutants (most pronounced for P36A and R38A). With both receptors, many mutants lost inhibitory activity, but the increased inhibition was observed for P36A at α7-GlyR. Thus, there are subtype-specific and common ws-LYNX1 residues recognizing distinct targets. Because ws-LYNX1 was inactive against glycine receptor, its “non-classical” binding sites on α7 nAChR should be within the extracellular domain. Micromolar affinities and fast washout rates measured for ws-LYNX1 and its mutants are in contrast to nanomolar affinities and irreversibility of binding for α-bungarotoxin and similar snake α-neurotoxins also targeting α7 nAChR. This distinction may underlie their different actions, i.e. nAChRs modulation versus irreversible inhibition, for these two types of three-finger proteins.     

Rare species of cyclostomata and fish in the Upper Oka basin and within Kaluga oblast

The species list of fish of the Upper Oka basin is verified. Data on biology of rare species are presented (western brook lamprey Lampetra planeri, sterlet Acipenser ruthenus, bystryanka Alburnoides bipunctatus rossicus, asp Aspius aspius, undermouth Chondrostoma variabile, bitterling Rhodeus sericeus, and fresh-water sculpin Cottus gobio koshewnikowi). Their morphological characters, distribution in the Upper Oka basin relative abundances of these species, and its changes during recent decades are also commented.     

Does rotan <Emphasis Type="Italic">Perccottus glenii</Emphasis> (Perciformes: Odontobutidae) eat the eggs of fish and amphibians?

The introduced fish rotan Perccottus glenii negatively affects the native ecosystems. An integrated study was performed to verify the suggestion that P. glenii feeds on eggs of fish (Rutilus rutilus, Abramis brama, Perca fluviatilis) and amphibians (Rana temporaria, R. arvalis, Bufo bufo). It was found that, during the intraoral testing of fish eggs, P. glenii perceives it as an edible item and swallows; however, under experimental conditions, this predator is limited in its ability for finding this prey because of its immobility. At the last stages of development, the movements of embryos within eggs attract the attention of P. glenii. This increases the vulnerability of fish eggs. In a large water body, there may be a spatial biotopic separation of the sites of egg deposition of some fish (for instance, perch) and the sites of concentrations of P. glenii. This makes eggs inaccessible for this predator. The amphibian eggs, on the contrary, are rejected by P. glenii after intraoral testing, which is determined by the presence of a thick jelly coat. During the period of development of eggs of frogs and toad in a pond, P. glenii actively feed (diverse invertebrates and adult newts were found in their diet); however, they ignore amphibian eggs that successfully develop in water bodies inhabited by P. glenii.     

Photodynamic effect of deuteroporphyrin IX and hematoporphyrin derivatives on single neuron

The photodynamic effects of 6 new deuteroporphyrin IX derivatives with different amphiphilicity and lipophilicity, as well as effects of known hematoporphyrin derivatives Photofrin II and Photoheme on isolated crayfish mechanoreceptor neurons were studied. After 30 min photosensitization, neurons were irradiated with He-Ne laser (632.8 nm, 0.3 W/cm(2)), and changes in their firing frequency were recorded. Neuron firing was shown to be very sensitive to photodynamic effect of the studied deuteroporphyrin IX derivatives causing irreversible firing abolition at pikomolar concentrations while Photoheme and Photofrin II were effective in the nanomolar range. The most effective sensitizers were 4-(1-methyl-3-hydroxybutyl)- and 4-(1-methyl-2-acetyl-3-oxobutyl)-deuteroporphyrins. Extinction and amphiphilicity were shown to be the most important properties determining photodynamic efficiency of the studied photosensitizers.     

Rocefin (ceftriaxone) in therapy of uncomplicated gonorrhea in males]

One hundred and nineteen males were examined in a unit for anonymous examination and treatment of sexually transmitted infections and the diagnosis of gonorrhea as monoinfection was stated. Rocefin (ceftriaxine) in a single dose of 250 mg intravenously was prescribed. The control examinations were performed in I and 3 weeks with microscopic and cultural methods. The clinical signs ceased by the 1st or 2nd days. The efficacy of the clinical and microbiological cure was 100%. Ceftriaxone proved to be an optimal agent for the treatment of uncomplicated gonorrhea in males.