Kinetic isotope effect and the presteady-state kinetics of the reaction catalyzed by the bacterial formate dehydrogenase

The primary kinetic isotope effect of the reaction catalyzed by NAD+-dependent formate dehydrogenase (EC 1.2.1.2.) from the methylotrophic bacterium Pseudomonas sp. 101 has been studied. Analysis of the ratios HVm/DVm and H(Vm/KM)/D(Vm/KM) in the pH range 6.1-7.9 showed that the transfer of hydride ion in ternary enzyme-substrate complex is a limiting step of the reaction, and the formate binding to the binary complex (formate dehydrogenase + NAD+) reached equilibrium when the pH of the medium was increased. An approach has been developed to determine the elementary constants of substrate association (kon) and dissociation (koff) at the stages of the binary--ternary enzyme-substrate complexes for the random equilibrium 2-substrate kinetic mechanism. The kon and koff values obtained for the bacterial formate dehydrogenase by using the proposed approach for NAD+ were (4.8 +/- 0.8)*10(5)M-1s-1 and (90 +/- 10) s-1, and for formate (2.0 +/- 1.0)*10(4) M-1s-1 and (60 +/- 20) s-1, respectively.     

Archaea in boreal Swedish lakes are diverse,dominated by Woesearchaeota and follow deterministic community assembly

Despite their key role in biogeochemical processes, particularly the methane cycle, archaea are widely underrepresented in molecular surveys because of their lower abundance compared with bacteria and eukaryotes. Here, we use parallel high-resolution small subunit rRNA gene sequencing to explore archaeal diversity in 109 Swedish lakes and correlate archaeal community assembly mechanisms to large-scale latitudinal, climatic (nemoral to arctic) and nutrient (oligotrophic to eutrophic) gradients. Sequencing with universal primers showed the contribution of archaea was on average 0.8% but increased up to 1.5% of the three domains in forest lakes. Archaea-specific sequencing revealed that freshwater archaeal diversity could be partly explained by lake variables associated with nutrient status. Combined with deterministic co-occurrence patterns this finding suggests that ecological drift is overridden by environmental sorting, as well as other deterministic processes such as biogeographic and evolutionary history, leading to lake-specific archaeal biodiversity. Acetoclastic, hydrogenotrophic and methylotrophic methanogens as well as ammonia-oxidizing archaea were frequently detected across the lakes. Archaea-specific sequencing also revealed representatives of Woesearchaeota and other phyla of the DPANN superphylum. This study adds to our understanding of the ecological range of key archaea in freshwaters and links these taxa to hypotheses about processes governing biogeochemical cycles in lakes.     

Comparison of an iterative microcalorimetric method and dialysis equilibrium for calculating thermodynamic parameters of a binding protein which presents a weak affinity for its substrate

An original iterative microcalorimetric method is used to study the interaction of 5-fluorouracil, a cancer chemotherapeutic agent, with human serum albumin. Two equivalent binding sites are demonstrated with an association constant and an enthalpy variation equal to 370 ± 30 m^?1 and ?10 ± 0,5 kcal/mol, respectively. By means of a competitive microcalorimetric method, the partially competitive binding of dipotassium chlorazepate and 5-fluorouracil is shown. This iterative microcalorimetric method can be applied to all techniques involving the measurement of a phenomenon which is proportional to the concentration of the complex.     

Discovery of Trunk Coelomoducts in Hemichordata

Histological examination of a specimen of a deep-sea enteropneusts that belongs to a yet undescribed species (Torquaratoridae gen. sp.) revealed numerous trunk coelomoducts. They open into the genital wing coelom as a typical funnels; short ducts communicate with environment through pores located on the outer side of the genital wings. Total number of coelomoducts in a specimen is estimated at several thousand. Trunk coelomoducts have not been found earlier in any member of the phylum. We believe that the release of the male gonad products occurs through coelomoducts of Torquaratoridae gen. sp.     

Investigation of ribosomal RNAs from liver mitochondria of albino rats]

Study of sedimentation coefficients of rat liver mt-rRNAs labelled in vivo under conditions of selective inhibition by actinomycin D of nuclear RNAs biosynthesis, in hypoosmic system of isolated mitochondria, and of mt-rRNAs isolated from purified 55 ribosomes revealed that mt-rRNAs are represented by 16S and 12S components. Both components are found to be of AU-type. Anomalous behaviour of mtRNA on MAK columns is observed. Some aspects of isolation and identification of mt-rRNAs in higher organisms are discussed.     

Morphogenesis of the genital system of Passerilepsis crenata (Cestoidea: Hymenolepididae)

The consecutive development of the genital organs in proglottides of the mature st obile of Passerilepis crenata was studied. The organs of the male and female genital systems, excluding testes and efferent ductules, are laid as a common genital anlage, which later become subdivided into three departments. On the basis of the degree of development of the genital organs zonation of the worm's strobile has been specified.