Soy and Rapeseed Protein Hydrolysis by the Enzyme Preparation Protosubtilin

A comparative study of soybean and rapeseed protein hydrolysis by protosubtilin, an original Russian enzyme preparation widely used in animal feed production, has been performed. SDS-PAG electrophoresis, HPLC, and mass spectrometry have been employed to analyze the obtained products. The soybean protein isolate used for hydrolysate production was obtained from a commercial supplier, and rapeseed proteins were prepared from the meal by alkali extraction. Low molecular weight impurities were removed by ultrafiltration. The degree of protein hydrolysis has been shown to depend on the substrate-to-enzyme preparation ratio, hydrolysis time, and protein concentration. Rapeseed protein hydrolysis by protosubtilin at an enzyme/protein ratio of 1: 20 and hydrolysis time of 20 h resulted in complete cleavage of the proteins present in the raw material and the accumulation of oligopeptides (molecular weight less than 14 kDa) and free amino acids, which accounted for 53 and 8% of the initial protein weight, respectively. In contrast to rapeseed proteins, soybean proteins showed considerable gelling at the initial stages of hydrolysis, and the formation of insoluble hydrolysis-resistant fragments was observed. The soluble part of the hydrolysate contained short oligopeptides and free amino acids, which accounted for 13% of the initial protein weight only.     

Recruitment of TBK1 to cytosol‐invading Salmonella induces WIPI2‐dependent antibacterial autophagy

Mammalian cells deploy autophagy to defend their cytosol against bacterial invaders. Anti‐bacterial autophagy relies on the core autophagy machinery, cargo receptors, and “eat‐me” signals such as galectin‐8 and ubiquitin that label bacteria as autophagy cargo. Anti‐bacterial autophagy also requires the kinase TBK1, whose role in autophagy has remained enigmatic. Here we show that recruitment of WIPI2, itself essential for anti‐bacterial autophagy, is dependent on the localization of catalytically active TBK1 to the vicinity of cytosolic bacteria. Experimental manipulation of TBK1 recruitment revealed that engagement of TBK1 with any of a variety of Salmonella‐associated “eat‐me” signals, including host‐derived glycans and K48‐ and K63‐linked ubiquitin chains, suffices to restrict bacterial proliferation. Promiscuity in recruiting TBK1 via independent signals may buffer TBK1 functionality from potential bacterial antagonism and thus be of evolutionary advantage to the host.     

Indication and identification of Lactobacillus spp. with the use of polymerase chain reaction

AIM: The aim of the work is the development of laboratory test for indication and identification of Lactobacillus spp. by the polymerase chain reaction. MATERIALS AND METHODS: The work is developed on the base of the GenBank/EMBL data about genetic sequences of the Lactobacillus spp. The sequences of DNA were studied with the help of the ClustalW program. The strains of the Lactobacillus spp., which are the object of the research, have been registered in Russian collection of industrial microorganisms. RESULTS: The laboratory test of nested-PCR for indication and identification L. plantarum, L. fermentum, L. acidophilus, L. delbrueckii, L. casei, L. rhamnosus was performed. The specificity of the nested-PCR was correlated with the control analyses of monoculture Lactobacillus spp. and commercial products. CONCLUSION: The new developed laboratory nested-PCR test may be use in control system of milk foods enriched by probiotic microorganisms.     

Production of alloplasmic and euplasmic wheat-barley ditelosomic substitution lines 7H<Superscript>1</Superscript>L<Superscript>mar</Superscript>(7D) and analysis of the 18S/5S mitochondrial repeat in these lines

Alloplasmic lines of common wheat with disomic substitution of chromosome 7D for telocentric chromosome 7H¹L^mar of barley H. marinum subsp. gussoneanum Hudson were isolated from the plants of generation BC₃, produced as a result of backcrossing of barley-wheat hybrids H. marinum subsp. gussoneanum (2n = 28) × T. aestivum (2n = 42), Pyrotrix, cultivar, with 28 common wheat cultivars Pyrotrix 28 and Novosibirskaya 67. Chromosome substitution pattern was determined using SSR analysis and C-banding. In preliminary genomic in situ hybridization experiments, telocentric chromosomes were assigned to wild barley was established. In the BC₃F₈ generations of three alloplasmic lines with the 7H¹L^mar(7D) substitution type the differences in fertility manifestation were observed: most of the L-32(1) plants were sterile, in line L-32(2) only sporadic plants were sterile, and line L-32(3) was fertile. Simultaneously with these experiments, using selfpollinated progeny of the hybrids obtained in crosses of common wheat cultivar Saratovskaya 29 (2n = 41), monosomic for chromosome 7D, with common wheat cultivar Pyrotrix 28 with addition of pair of telocentric chromosomes 7H¹L^mar (7D) of barley H. marinum subsp. gussoneanum, euplasmic wheat-barley ditelosomic substitution 7H¹L^mar (7D) lines were isolated. The lines obtained had normal fertility. PCR analysis of the 18S/5S mitochondrial repeat (hereafter, mtDNA sequence) in alloplasmic and euplasmic ditelosomic substitution lines 7H¹L^mar(7D) was performed. In the plants from alloplasmic sterile line L-32(1), the sequences only of the barley (maternal) type were revealed, while the plants from alloplasmic fertile lines L-32(2) and L-32(3) demonstrated heteroplasmy (the presence of barley- and wheat-like sequences within one individual). In euplasmic ditelosomic substitution lines the presence of only wheat-like 18S/5S mitochondrial repeat sequences was observed. The results indicate that the presence of barley-like mtDNA sequences in alloplasmic substitution lines was not associated with the presence of barley chromosomes in their nuclear genomes.     

Codon usage bias and the evolution of influenza A viruses. Codon Usage Biases of Influenza Virus

Background  

The influenza A virus is an important infectious cause of morbidity and mortality in humans and was responsible for 3 pandemics in the 20^th century. As the replication of the influenza virus is based on its host's machinery, codon usage of its viral genes might be subject to host selection pressures, especially after interspecies transmission. A better understanding of viral evolution and host adaptive responses might help control this disease.     

Bacteria of the genus Burkholderia as a typical component of the microbial community of Sphagnum peat bogs

Bacteria of the genus Burkholderia are a typical component of the microbial complex of Sphagnum peat bogs and constitute a substantial portion of the aerobic chemoorganotrophic isolates which are routinely obtained from these environments on an acidic nutrient media. The ecophysiological characteristics of the 27 strains of such organisms, which were isolated from the peat of acidic Sphagnum bogs of the boreal and tundra zones of Russia, Canada, and Estonia, were investigated in the present study. Most of the Burkholderia strains isolated from these bogs were phylogenetically close to the species B. glathei, B. phenazinium, B. fungorum, and B. caryophylli, the typical inhabitants of soil and plant rhizosphere. The bog isolates utilized a broad range of substrates as carbon and energy sources, including organic acids, sugars, polyalcohols, and certain aromatic compounds. All the strains studied were capable of growth on nitrogen-free media. They developed in the pH range of 3.5 to 7.4 and from 3 to 37°C, with the optima at pH 5–7 and 11–23°C, respectively. They were therefore moderately acidophilic, psychroactive, dinitrogen-fixing microorganisms well adapted to the conditions of acidic northern Sphagnum bogs.     

Immunohistochemical investigation of Bcl-2 and p53 levels in rat hypothalamus after sleep deprivation

On Wistar rats in view of electrophysiological parameters after sleep deprivation (SD; awake by gentle handling method) and the subsequent postdeprivative sleep (PDS) immunohistochemical investigation of Bcl-2 and p53 peptides optical density levels in neurons of paraventricular (PVN), supraoptic (SON) and median (MnPN) hypothalamus nuclei was carried out. The Bcl-2 was increased in all nuclei both after SD and PDS. The level of p53 was increased in PVN and SON after SD and PDS, but in MnPN only on PDS. Any morphological attributes of apoptosis in the nuclei was not revealed. Obtained data testify an active role of p53 and Bcl-2 peptides in regulation of neuronal activity in hypothalamus at change of a cycle wakefulness-sleep.     

Functional metabolic characteristics of sodium hyaluronate in mammal organism

In the review data concerning functional metabolic characteristics of sodium hyaluronate in mammal organism, as well as autologous results concerning the study of its intraarticular action on metabolism of conjunctive tissue in the conditions of experimental arthritis in rabbits are given.     

Links between plant and fungal communities across a deforestation chronosequence in the Amazon rainforest

Understanding the interactions among microbial communities, plant communities and soil properties following deforestation could provide insights into the long-term effects of land-use change on ecosystem functions, and may help identify approaches that promote the recovery of degraded sites. We combined high-throughput sequencing of fungal rDNA and molecular barcoding of plant roots to estimate fungal and plant community composition in soil sampled across a chronosequence of deforestation. We found significant effects of land-use change on fungal community composition, which was more closely correlated to plant community composition than to changes in soil properties or geographic distance, providing evidence for strong links between above- and below-ground communities in tropical forests.