Respiratory heat loss of Holstein cows in a tropical environment

In order to develop statistical models to predict respiratory heat loss in dairy cattle using simple physiological and environmental measurements, 15 Holstein cows were observed under field conditions in a tropical environment, in which the air temperature reached up to 40°C. The measurements of latent and sensible heat loss from the respiratory tract of the animals were made by using a respiratory mask. The results showed that under air temperatures between 10 and 35°C sensible heat loss by convection decreased from 8.24 to 1.09 W m^–2, while the latent heat loss by evaporation increased from 1.03 to 56.51 W m^–2. The evaporation increased together with the air temperature in almost a linear fashion until 20°C, but it became increasingly high as the air temperature rose above 25°C. Convection was a mechanism of minor importance for respiratory heat transfer. In contrast, respiratory evaporation was an effective means of thermoregulation for Holsteins in a hot environment. Mathematical models were developed to predict both the sensible and latent heat loss from the respiratory tract in Holstein cows under field conditions, based on measurements of the ambient temperature, and other models were developed to predict respiration rate, tidal volume, mass flow rate and expired air temperature as functions of the ambient temperature and other variables.This paper forms part of A. S. Campos Maias doctoral thesis.     

Stability Study of Crude Dextransucrase from <Emphasis Type="Italic">Leuconostoc citreum</Emphasis> NRRL B-742

In the present work, the stability of crude dextransucrase from Leuconostoc citreum B-742 was evaluated in synthetic and in cashew apple juice culture broth. Optimum stability conditions for dextransucrase from L. citreum B-742 were different from the reported for its parental industrial strain enzyme (L. mesenteroides B-512F). Crude dextransucrase, from L. citreum B-742, produced using cashew apple juice as substrate, presented higher stability than the crude enzyme produced using synthetic culture medium, showing the same behavior previously reported for dextransucrase from L. mesenteroides B-512F. The crude enzyme presented good stability in cashew apple juice for 48 h at 25°C and pH 6.5.     

Ab initio structure of human seminal plasma prostatic inhibin gives significant insight into its biological functions

Human seminal plasma prostatic inhibin (HSPI) is a protein isolated from the human prostate gland. Despite its profound biomedical and biotechnological importance, the 3D structure of this protein of 94 amino acids remains undeciphered. The difficulties in extracting it in pure form and crystallizing it have restrained the determination of its structure experimentally. The homology-based computational methods are also not applicable, as HSPI lacks sufficient sequence homology with known structures in the protein data banks. We have predicted the structure of HSPI by a knowledge-based method using nonparametric multivariate statistical techniques. Stereochemical and other standard validation tests confirm this to be a well-refined structure. The biophysical properties exhibited by this structure are in good agreement with the NMR experimental observations. Docking and other computational studies on this structure provide significant explanation and insight into its binding activities and related biological and immunogenic functions and offer new directions for its potential applications.     

A protein-free diet changes synaptosomal membrane fluidity and tyrosine and glutamate transport

Synaptosomes were isolated from cerebrums of rats fed standard (20% protein) or protein-free diets for 30 days. Arrhenius plots of their (Na⁺/K⁺)ATPase activities revealed a transition temperature of 25.5°C for control rats and 23.4°C for rats on protein-free diet, indicating that the latter increases synaptosomal membrane fluidity. The only change observed in the composition of the synaptosomal membranes was a 26% decrease of sialic acid. In synaptosomes from rats on protein-free diet the uptake of tyrosine was slightly reduced while that of glutamate was not affected. However, the exit of glutamate was reduced.     

Paleolimnological reconstruction of the trophic state in Lake Balaton (Hungary) using Cladocera remains

The sediment of Lake Balaton (Hungary) provides important information about the lake’s history, particularly with regard to eutrophication. In this study, we used fossil pigment analysis and subfossil Cladocera remains preserved in a dated sediment core to identify trophic stages from ~250 bc to present. Dates of the most recent eutrophic events are in good agreement with previously published data. In general, the abundance and diversity of the Cladocera community increased with eutrophication and decreased with oligotrophication. The sediments of Lake Balaton were characterised by Chydoridae remains, of which Alona species were the most abundant. Of these, Alona quadrangularis and Alona affinis accounted for 40 and 20% of the total Cladocera remains, respectively. The trophic state of Lake Balaton varied between mesotrophic and eutrophic regimes. Seven different trophic periods were identified in Lake Balaton on the basis of Sedimentary Pigment Degradation Unit (SPDU) content of the sediment. Eutrophic states were (1) from ~250 to ~30 bc, (3) between ~300 and ~590 ad, (5) between 1834 and 1944 and (7) from the 1960s until present. Mesotrophic states were (2) ~30 bc to ~300 ad, (4) 590–1834, (6) 1944–1960s. Discriminant analysis of the cladoceran data confirmed these historic events, except for the short mesotrophic episode between 1944 and 1960. The first stage of eutrophication of Lake Balaton (~250 to ~30 bc) was characterised by extensive macrophyte vegetation, as indicated by the increasing abundance of vegetation-associated Cladocera species (Eurycercus lamellatus, Sida crystallina, Pleuroxus sp.). Intensification of eutrophication was identified since the 1980s, reflected by a high abundance of Bosmina species. The most significant planktivorous fish of Lake Balaton was the Sabre carp (Pelecus cultratus), and when its number decreased, the abundance of Bosmina species increased. This study shows that Cladocera are responsive to trophic state changes, underlining their importance as a tool for the assessment of lake eutrophication.     

Purification and characterization of the FeII- and alpha-ketoglutarate-dependent xanthine hydroxylase from Aspergillus nidulans

His6-tagged xanthine/alpha-ketoglutarate (alphaKG) dioxygenase (XanA) of Aspergillus nidulans was purified from both the fungal mycelium and recombinant Escherichia coli cells, and the properties of the two forms of the protein were compared. Evidence was obtained for both N- and O-linked glycosylation on the fungus-derived XanA, which aggregates into an apparent dodecamer, while bacterium-derived XanA is free of glycosylation and behaves as a monomer. Immunological methods identify phosphothreonine in both forms of XanA, with phosphoserine also detected in the bacterium-derived protein. Mass spectrometric analysis confirms glycosylation and phosphorylation of the fungus-derived sample, which also undergoes extensive truncation at its amino terminus. Despite the major differences in the properties of these proteins, their kinetic parameters are similar (kcat = 30-70 s-1, Km of alphaKG = 31-50 muM, Km of xanthine approximately 45 muM, and pH optima at 7.0-7.4). The enzyme exhibits no significant isotope effect when [8-2H]xanthine is used; however, it demonstrates a 2-fold solvent deuterium isotope effect. CuII and ZnII potently inhibit the FeII-specific enzyme, whereas CoII, MnII, and NiII are weaker inhibitors. NaCl decreases the kcat and increases the Km of both alphaKG and xanthine. The alphaKG cosubstrate can be substituted with alpha-ketoadipate (9-fold decrease in kcat and 5-fold increase in the Km compared to those of the normal alpha-keto acid), while the alphaKG analogue N-oxalylglycine is a competitive inhibitor (Ki = 0.12 muM). No alternative purines effectively substitute for xanthine as a substrate, and only one purine analogue (6,8-dihydroxypurine) results in significant inhibition. Quenching of the endogenous fluorescence of the two enzyme forms by xanthine, alphaKG, and DHP was used to characterize their binding properties. A XanA homology model was generated on the basis of the structure of the related enzyme TauD (PDB entry 1OS7) and provided insights into the sites of posttranslational modification and substrate binding. These studies represent the first biochemical characterization of purified xanthine/alphaKG dioxygenase.     

Chemical action of influenza virus on the red blood cell membrane. II. Action on membrane lipids and amino acids.

Haemagglutination by influenza virus strains PR8 and Lee as well as treatment of red blood cells by periodate produced numerous and manifold changes in the lipids and the amino acid composition of the erythrocytes. The changes induced by the influenza strains differed from each other and from those induced by periodate. Alterations in the membrane lipids observed after haemagglutination by heat treated strain PR8 and by its native form were identical.     

Experimental and theoretical study on high temperature induced changes in chlorophyll <Emphasis Type="Italic">a</Emphasis> fluorescence oscillations in barley leaves upon 2 % CO<Subscript>2</Subscript>

Oscillations in many of photosynthetic quantities with a period of about 1 min can be routinely measured with higher plant leaves after perturbation of the steady state by sudden change in gas phase. Among all hypotheses suggested so far to explain the oscillations, an effect of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) activation status to control the oscillations is highly probable, at least upon high temperature (HT) treatment when in vivo RuBPCO activity controlled by RuBPCO activase (RuBPCO-A) decreases. Therefore, we measured the oscillations in fluorescence signal coming from barley leaves (Hordeum vulgare L. cv. Akcent) after their exposure for various time intervals to different HTs in darkness. We also evaluated steady state fluorescence and CO₂ exchange parameters to have an insight to functions of electron transport chain within thylakoid membrane and Calvin cycle before initiation of the oscillations. The changes in period of the oscillations induced by moderate HT (up to 43 °C) best correlated with changes in non-photochemical fluorescence quenching (q_N) that in turn correlated with changes in gross photosynthetic rate (P _G) and rate of RuBPCO activation (k_act). Therefore, we suggest that changes in period of the oscillations caused by moderate HT are mainly controlled by RuBPCO activation status. For more severe HT (45 °C), the oscillations disappeared which was probably caused by an insufficient formation of NADPH by electron transport chain within thylakoid membrane as judged from a decrease in photochemical fluorescence quenching (q_P). Suggestions made on the basis of experimental data were verified by theoretical simulations of the oscillations based on a model of Calvin cycle and by means of a control analysis of the model.