Genetical analysis of rifampicin resistant mutants of E. coli K 12

Summary E. coli rifampicin-resistant (rif-r) mutants were divided into two conventional groups: A, resistant to 50–100g/ml of rifampicin both on complete and minimal medium and B, sensitive to these concentrations of rifampicin on minimal, but resistant on complete medium. RNA polymerase from rif-r-A mutants is resistant to high concentrations of rifampicin in vitro while the enzyme from rif-r-B mutants but slightly if at all differs from the wild-type enzyme in its response to the drug.All rif-r-A and rif-r-B mutants are closely linked and map between argH and thi on E. coli K 12 chromosome. We failed to isolate any rifampicin-resistant mutants which would map outside this region.     

A folding pathway for betapep-4 peptide 33mer: from unfolded monomers and beta-sheet sandwich dimers to well-structured tetramers.

It was recently reported that a de novo designed peptide 33mer, betapep-4, can form well-structured beta-sheet sandwich tetramers (Ilyina E, Roongta V, Mayo KH, 1997b, Biochemistry 36:5245-5250). For insight into the pathway of betapep-4 folding, the present study investigates the concentration dependence of betapep-4 self-association by using 1H-NMR pulsed-field gradient (PFG)-NMR diffusion measurements, and circular dichroism. Downfield chemically shifted alphaH resonances, found to arise only from the well-structured betapep-4 tetramer state, yield the fraction of tetramer within the oligomer equilibrium distribution. PFG-NMR-derived diffusion coefficients, D, provide a means for deriving the contribution of monomer and other oligomer states to this distribution. These data indicate that tetramer is the highest oligomer state formed, and that inclusion of monomer and dimer states in the oligomer distribution is sufficient to explain the concentration dependence of D values for betapep-4. Equilibrium constants calculated from these distributions [2.5 x 10(5) M(-1) for M-D and 1.2 x 10(4) M(-1) for D-T at 313 K] decrease only slightly, if at all, with decreasing temperature indicating a hydrophobically mediated, entropy-driven association/folding process. Conformational analyses using NMR and CD provide a picture where "random coil" monomers associate to form molten globule-like beta-sheet sandwich dimers that further associate and fold as well-structured tetramers. Betapep-4 folding is thermodynamically linked to self-association. As with folding of single-chain polypeptides, the final folding step to well-structured tetramer betapep-4 is rate limiting.     

Organization and evolution of bacterial and bacteriophage primase-helicase systems

Summary Amino acid sequences of primases and associated helicases involved in the DNA replication of eubacteria and bacteriophages T7, T3, T4, P4, and P22 were compared by computer-assisted methods. There are two types of such systems, the first one represented by distinct helicase and primase proteins (e.g., DnaB and DnaG proteins of Escherichia coli), and the second one by single polypeptides comprising both activities (gp4 of bacteriophages T7 and T3, and alpha protein of bacteriophage P4). Pronounced sequence similarity was revealed between approximately 250 amino acid residue N-terminal domains of stand-alone primases and the primase-helicase proteins of T7(T3) and P4. All these domains contain, close to their N-termini, a conserved Zn-finger pattern that may be implicated in template DNA recognition by the primases. In addition, they encompass five other conserved motifs some of which may be involved in substrate (NTP) binding. Significant similarity was also observed between the primase-associated helicases (DnaB, gp12 of P22 and gp41 of T4) and the C-terminal domain of T7(T3) gp4. On the other hand the C-terminal domain of P-alpha of P4 is related to another group of DNA and RNA helicases. Tentative phylogenetic trees generated for the primases and the associated helicases showed no grouping of the phage proteins, with the exception of the primase domains of bacteriophages T4 and P4. This may indicate a common origin for one-component primase-helicase systems. Two scenarios for the evolution of primase-helicase systems are discussed. The first one involves fusion of the primase and helicase components (T7 and T3) or fusion of the primase component with a different type of helicase domain (P4). The second possibility is the duplication of an ancestral gene encoding a gp4-like bifunctional protein followed by divergence of the copies, one of which retains the primase and the other the helicase domain.     

Novel and simple ELISA-based method for antibody affinity determination

New coordinates for antibody affinity determination by ELISA are suggested. The suggested approach is very simple but at the same time it is more convenient and is deprived of the drawbacks inhered in the earlier suggested methods. The examples of antibody affinity determination by the suggested methods for both simulative and experimental binding curves are considered. It was demonstrated that the suggested methods allow getting more precise values of antibody affinity.     

Antioxidant,Antimicrobial, and Fungicidal Properties of Chitosan Based Films (Review)

Data on the effect of biologically active compounds introduced into chitosan-based films (polyphenols, natural extracts from plants and proteins and peptides) on their antimicrobial, antioxidant and fungicidal activity are summarized and analyzed. It was shown that composite films reveal higher antimicrobial activity than films containing only chitosan, and they also retain their mechanical and protective properties.     

Differentiation of tetrapod communities and some aspects of biotic events in the early triassic of Eastern Europe

The patterns of spatial differentiation of the Early Mesozoic terrestrial biota in Eastern Europe and Australia-Tasmania demonstrate that the tetrapod faunal recovery following the Permian extinction was characterized by both global and regional heterogeneity. Local distinctions observed in the development of Early Triassic tetrapod assemblages of European Russia allow the recognition of the following realms: (1) the central and northern regions of the East European Platform (Moscow-Mezen Syncline) and the Timan-North Ural Region; (2) the southern Fore-Urals, including the Obshchii Syrt Plateau; and (3) the southern regions of the East European Platform (the slope of the Voronezh Anticline). Climatic conditions at the initial stage of the development of local communities were characterized by an increase in aridity and seasonal contrasts of climate. Therefore, terrestrial assemblages mostly concentrated in the aquatic and coastal biotopes. Accordingly, vertebrate assemblages of the region were dominated everywhere by aquatic amphibians and semiaquatic reptiles, while the accompanying palynomorph assemblages show the predominance of hygrophilous vegetation indicative of swampy mangrove setting. With respect to amphibians, a peak of local biogeographic differentiation falls on the onset of the Early Triassic and, in the case of reptiles, on the end of this time span. This change conforms to the increasing role of reptiles in the overall taxonomic diversity with time. Among the three main biogeographic units of the region, the Southern Fore-Ural Realm is distinguished by the maintenance of distinct faunal links with Gondwanan regions. The Southern Realm shows a connection with the Germanic Basin and more western Euramerican areas, which is documented for the Late Olenekian and occurred under influence of coastal marine conditions.     

Biofilms as a Mode of Existence of Bacteria in External Environment and Host Body: The Phenomenon,Genetic Control,and Regulation Systems of Development

Studies of the last decade have shown that most bacteria exist in natural ecosystems as specifically organized, attached to substrates biofilms rather than as freely floating plankton cells. The formation of these biofilms is a complex and highly regulated process. The development of biofilm communities is a primary strategy of bacterial survival not only in the external environment but also in the bodies of infected macroorganisms. In these organisms, bacteria are joined by complicated cell–cell associations, which makes them functionally similar to multicellular organisms. In the present review, we consider the structural organization of biofilms, factors affecting initiation of the biofilm formation, differential expression of bacterial genes at various stages of the biofilm development and their regulation. The significance of studies in this field for medicine, in particular, for prevention and protection against pathogenic bacteria, is discussed.     

Fatty acid ethyl esters in meconium: A biomarker of fetal alcohol exposure and effect

To determine if meconium fatty acid ethyl esters (FAEE) in rat pups is a good biomarker of prenatal exposure and effect to alcohol, three groups of pregnant rats were studied: one control (pair fed) and two treatment groups given 25% alcohol at 2.2 or 5.5 g⁻¹ kg⁻¹ d⁻¹. The pups were delivered on day 20 and, for each dam, were separated into a male and female group. The body, brain, intestines, and placenta of the pups were obtained, weighed, and stored at −20°C. The pups’ intestines (as surrogate of meconium) from each group were pooled, and meconium was analyzed by gas chromatography/mass spectroscopy for FAEE. The meconium showed the following FAEE: ethyl palmitate, ethyl stearate, and ethyl linolenate and were only found in the alcohol-treated group and with high specificity but low sensitivity. Mean body weight of the pups was lower in the treatment groups compared to the control groups. Ethyl palmitate concentration correlated negatively to the pups’ mean body and brain weights. Therefore, ethyl palmitate, stearate, and linolenate, in meconium of rat pups prenatally exposed to alcohol, are useful biomarkers of prenatal alcohol exposure, with ethyl palmitate a good biomarker of adverse effect on the pups’ body and brain weight.