Infant gut microbiome is enriched with Bifidobacterium longum ssp. infantis in Old Order Mennonites with traditional farming lifestyle.

BACKGROUND: Growing up on traditional, single-family farms is associated with protection against asthma in school age, but the mechanisms against early manifestations of atopic disease are largely unknown. We sought determine the gut microbiome and metabolome composition in rural Old Order Mennonite (OOM) infants at low risk and Rochester, NY urban/suburban infants at high risk for atopic diseases. METHODS: In a cohort of 65 OOM and 39 Rochester mother-infant pairs, 101 infant stool and 61 human milk samples were assessed by 16S rRNA gene sequencing for microbiome composition and qPCR to quantify Bifidobacterium spp. and B. longum ssp. infantis (B. infantis), a consumer of human milk oligosaccharides (HMOs). Fatty acids (FAs) were analyzed in 34 stool and human 24 milk samples. Diagnoses and symptoms of atopic diseases by 3 years of age were assessed by telephone. RESULTS: At a median age of 2 months, stool was enriched with Bifidobacteriaceae, Clostridiaceae, and Aerococcaceae in the OOM compared with Rochester infants. B. infantis was more abundant (p < .001) and prevalent, detected in 70% of OOM compared with 21% of Rochester infants (p < .001). Stool colonized with B. infantis had higher levels of lactate and several medium- to long/odd-chain FAs. In contrast, paired human milk was enriched with a distinct set of FAs including butyrate. Atopic diseases were reported in 6.5% of OOM and 35% of Rochester children (p < .001). CONCLUSION: A high rate of B. infantis colonization, similar to that seen in developing countries, is found in the OOM at low risk for atopic diseases.

Polycyclic aromatic hydrocarbons, antibiotic resistance genes, toxicity in the exposed to anthropogenic pressure soils of the Southern Russia.

The influence of anthropogenic pollution, particularly with polycyclic aromatic hydrocarbons (PAHs) on soil toxicity and spread of antibiotic resistance genes (ARGs) is extremely important nowadays. We studied 20 soil samples from a technogenically polluted site, municipal solid wastes (MSW) landfills, and rural settlements in the southwestern part of the Rostov Region of Russia. A close correlation was established between the results of biosensor testing for integral toxicity, the content of genes for the biodegradation of hydrocarbons, and the concentration of PAHs in soils. The relation between the quantitative content of ARGs and the qualitative and quantitative composition of PAHs has not been registered. Soils subjected to different types of the anthropogenic pressure differed in PAHs composition. The technogenic soils are the most polluted ones. These soils are enriched with 5 ring PAHs and carry the maximum variety of assayed ARGs, despite the fact that they do not receive household or medical waste.

The impact of length variations in the L2 loop on the structure and thermal stability of non-specific porins: The case of OmpCs from the Yersinia pseudotuberculosis complex.

Porins are integral proteins of the outer membranes of gram-negative bacteria. In membranes, they exist as homotrimers and the L2 loops contribute to their stability. Comparison of OmpC porins of the Yersinia pseudotuberculosis complex with other enterobacterial porins demonstrated L2 loop length diversity, which is caused by varying numbers of dipeptide/tripeptide repeats. The OmpC porins are highly homologous to each other, and they can be subdivided into five isoforms based on their L2 loop structure. Optical spectroscopy and SDS-PAGE experiments revealed that particularities of the L2 loops affected the structure and thermal stability of the porins. Thermal denaturation studies showed that porins with shorter loops, compared to porins with longer loops, had more stable tertiary and less stable secondary and quaternary structures. According to our comparative modeling results, the L2 loops differ in their structure by adopting different spatial positions and forming different polar bonds with a neighbor monomer. The replacement of asparagine with arginine at the C-terminus of the L2 loop shifts the loop upwards and causes the loss of contacts with the arginine clusters within the pores. The increase in the length of these loops ensures that they shift down toward the pore and restore their contacts with arginines on the channel wall, as is the case in classical nonspecific porins. Despite the fact that the surface charge density varies considerably among the OmpC porins, the L2 loops form a typical negatively charged region in the center of the trimer.

[Regulation of Yersinia pseudotuberculosis major porin expression in response to antibiotic stress].

The OmpF porin gene expression in Yersinia pseudotuberculosis in response to antibiotics of two different classes (kanamycin and nalidixic acid) was analyzed using quantitative PCR and a fluorescence reporter system. Both antibiotics downregulated the expression of the ompF gene. The nalidixic acid significantly reduced ompF expression, while kanamycin, for which porins are considered to be an alternative transport route, only slightly reduced the ompF level.

Pollution impact on microbial communities composition in natural and anthropogenically modified soils of Southern Russia.

Metagenomic studies of soil microbocenoses are extremely relevant nowadays. The study of pollution impact on soil microbiomes is of particular interest. The structure of microbial communities in soils with different levels of pollution by polycyclic aromatic hydrocarbons (PAHs) and potentially toxic elements (PTEs) was studied. High bacterial biodiversity was found in all the studied soil samples, but its lowest values are found in soil samples taken on the territory of technogenically polluted Lake Atamanskoye. Assessment of soil pollution showed the highest content of polycyclic aromatic hydrocarbons (PAHs) and potentially toxic elements (PTEs) for the soils Lake Atamanskoye. The high content of pollutants negatively affects the abundance of representatives of the phyla Actinobacteria, Planctomycetes, Verrucomicrobia, and Nitrospirae. Such phyla as Proteobacteria, Candidate Divisions TM7, OD1, WPS-2, Chlamydiae, Cyanobacteria are characterized by positive direct correlation with the content of pollutants, especially with PAHs. A cooperative effect of decrease in the number of Actinobacteria and Proteobacteria with an increase in Armatimonadetes probably corresponds to PTEs contamination. The proportion of Candidate Division OD1, Chlamydiae, Cyanobacteria, and Candidate Division WPS-2 was increased in the soil microbiome under the influence of severe combined pollution. Pollutants negatively affect the abundance of dominant unclassified_o__Gaiellales and unclassified_o__WD2101 genera. Iamia, Salinibacterium, Arthrobacter, Kaistobacter, Thiobacillus genera are characterized by a low abundance, but they are presumably the most resistant to soil pollution. It was revealed that the level of soil pollution largely determines the composition and diversity of bacterial communities in the soils of the studied territories. Operating taxonomic units have been established that have prognostic value for assessing the state, level of soil pollution, and their biological safety.

Evidence for using pimavanserin for the treatment of Parkinson's disease psychosis.

The aim of this editorial is to evaluate the evidence for using pimavanserin for the treatment of Parkinson's disease psychosis (PDP) from randomized controlled trials (RCTs). We only identified two published trials that evaluated the use of pimavanserin among individuals with PDP. Both studies found that pimavanserin improved psychotic symptoms among individuals with PDP when compared to placebo. Pimavanserin was fairly well tolerated in both studies and did not appear to cause significant sedation or worsen motor symptoms among individuals with PDP. However, given the limited data, additional confirmatory studies are required before pimavanserin can be considered as a first line agent for the treatment of psychotic symptoms among individuals with PD.

Virulence-associated fyuA/irp2 gene cluster of Yersinia enterocolitica biotype 1B carries a novel insertion sequence IS1328.

The fyuA/irp2 gene cluster, which is part of the Yersinia pestis pigmentation (pgm) locus encoding genes involved in iron uptake and virulence, is present in all pesticin-sensitive bacteria. In Y. enterocolitica biotype 1B strains (serotypes O8, O20, O21), the fyuA/irp2 gene cluster carries an insertion of a novel repetitive sequence, IS1328. It was also found in the genome of Y. enterocolitica O5 (biotype 1A) and O13 (biotype 1B), but not in pesticin-sensitive Y. pseudotuberculosis O1 and Escherichia coli Phi. The 1353-bp repetitive element has 12-bp perfect inverted terminal repeats. A single open reading frame is capable of encoding a 334-amino acid polypeptide. IS1328 DNA has high homology with the DNA sequences located downstream of the aggR gene from the enteroaggregative E. coli (EAggEC), to the region of the R751 plasmid flanking Tn501, to the sequence following the merR gene of S. marcescens pDU1358 plasmid, and to the sequences of K. pneumoniae plasmid pCFF04. The putative polypeptide has 36.4% identity with the transposase encoded by the Coxiella burnetii IS1111a insertion sequence. The IS1328 insertion sequence could be responsible for the deletions of the fyuA/irp2 gene cluster observed in Y. enterocolitica O8 and could represent a member of a new group of widely distributed repetitive elements.

Representational difference analysis uncovers a novel IS10-like insertion element unique to pathogenic strains of Yersinia enterocolitica.

The method of suppressive subtractive hybridization was employed to map out genomic differences between the highly pathogenic Yersinia enterocolitica (Ye) biogroup 1B, serotype O:8 strain (WA-314) and the closely related apathogenic Y. enterocolitica biogroup 1A, serotype O:5 strain (NF-O). A novel IS10-like element, IS1330, uncovered by this technique was found to be uniquely present in high copy numbers among the highly pathogenic Y. enterocolitica 1B strains, while a single copy of the element was found in the low pathogenic Ye biogroup 4 serotype O:3 strain. The 1321-bp repetitive element has 19-bp imperfect inverted terminal repeats and is bracketed by a 10-bp duplication of the target sequence. The predicted transposase shares high homology with the IS10 open reading frame of the large virulence plasmid pWR501, of Shigella flexneri, with IS10 transposase of Salmonella typhi, and with IS1999 (tnpA) of Pseudomonas aeruginosa. The IS1330 tnp gene is transcribed in vitro and in vivo in HeLa cells. At least one copy of IS1330 flanks the recently described chromosomal type III secretion cluster in Y. enterocolitica WA-314, O:8, and future studies should shed light on whether this novel transposase mediates transposition events in highly pathogenic Y. enterocolitica strains, thus enhancing the genetic plasticity of this species.

Characterization of a novel unique restriction-modification system from Yersinia enterocolitica O:8 1B.

Genetic manipulations with enteropathogenic Yersinia enterocolitica O:8 are complicated by the presence of an efficient PstI-like YenI restriction-modification (R-M) system. We have characterized the YenI R-M system in Y. enterocolitica O:8, biotype 1B. A 5039 bp DNA fragment of the pSAK2 recombinant plasmid carrying the yenI locus was used to determine the nucleotide sequence. DNA sequence analysis identified a single 2481 bp open reading frame (ORF) that encodes an 826 amino acid large polypeptide having an apparent molecular mass of 93 kDa. The N-terminal part of the YenI ORF has 45 and 40% identity to PstI and BsuI methyltransferases (MTases), respectively; while the C-terminal part depicts 55 and 45% identity to endonucleases (ENases) of both isoschyzomeric enzymes. The yenI gene was cloned into pT7-5 plasmid and has been shown to encode a single polypeptide of expected molecular mass. A specific recognition sequence, typical to the type II R-M systems and single peptide organization, typical to type IV R-M systems, make YenI unique among known restriction-modification systems. We have constructed a truncated recombinant variant of YenI enzyme, which conserved only MTase activity, and that can be applied to YenI methylation of the DNA to be transformed into Y. enterocolitica O:8 biotype 1B strains.

Local hopping of IS3 elements into the A+T-rich part of the high-pathogenicity island in Yersinia enterocolitica 1B, O:8.

The high-pathogenicity island (Yen HPI) of Yersinia enterocolitica biogroup (BG) 1B strains is associated with mouse virulence. Three repeated sequences are clustered on the A+T-rich part of the Yen HPI downstream of the fyuA yersiniabactin receptor gene in Y. enterocolitica O:8 strains WA-314 and 8081. In addition to IS1328 and IS1400, the RS3 repeated sequence consists of a novel insertion sequence, IS1329, inserted into the remnants of IS1222. This partial IS retains both 44-bp inverted terminal repeats (ITRs) of IS1222 but has suffered deletions of different sizes in strains WA-314 and 8081. IS1329 is 1243-bp long, carries 25-bp imperfect ITRs and two consecutive orfs capable to encode 110-amino acid (aa) and 249-aa proteins, respectively. IS1329 is present only in BG 1B Y. enterocolitica strains. Similarly to IS1400, IS1329 and IS1222 belong to the IS3 group of mobile elements and seem to have preference for the 'local hopping' into the A+T-rich part of the Yen HPI. These insertion sequences may be responsible for the imprecise deletions of the Yen HPI in strain WA-314.

Nucleotide sequence and structural organization of Yersinia pestis insertion sequence IS100.

Insertion sequence IS100 was localized on a 9.5-kb plasmid of Yersinia pestis and was shown to be specific for Y. pestis and serotype I strains of Y. pseudotuberculosis. The nucleotide sequence of IS100 isolated from this plasmid was determined. The element, which was flanked by 5-bp direct repeats, contained 1953 bp including imperfect inverted terminal repeats of 52 and 61 bp long (43 bp were identical). Two open reading frames encoding potential polypeptides of 340 and 252 amino acids were identified on one DNA strand. Nucleotide sequence as well as deduced polypeptide sequences of IS100 were homologous to those of IS21, IS232 and IS640.

Characterization of the integration site of Yersinia high-pathogenicity island in Escherichia coli.

The high-pathogenicity island (HPI) of virulent Yersiniae consists of (i) a functional core encoding for biosynthesis and uptake of the siderophore yersiniabactin and (ii) a 5- to 13-kb AT-rich region of unknown function. This Yersinia HPI has been shown to be widely distributed among different pathotypes of Escherichia coli. In this study, the insertion site of the HPI was defined in three different E. coli strains: The enteroaggregative E. coli (EAggEC) strain 17-2, the uropathogenic (UPEC) E. coli strain 536, and the probiotic E. coli DSM6601. We demonstrated that in all three E. coli isolates the HPI is associated with the asnT tRNA (5'-extremity) and truncated in the AT-rich region (3'-extremity) since the 17-bp direct repeat (DR) of the asn tRNA that flanks the HPI in Yersinia is missing in E. coli. Moreover, in comparison to the HPI-negative E. coli K-12 strain, a uniform deletion must have taken place in the E. coli chromosome adjacent to the 3'-border of the HPI.

Somatostatin-14 alters the thymus size and relation among the thymocyte subpopulations in peripubertal rats.

It is well known that somatostatin exerts a wide range of effects in the body, and acts as an autocrine or paracrine factor in the thymus. However, it has not been investigated yet whether somatostatin alters the thymus size and relation among the thymocyte subpopulations in the peripubertal rats. For this purpose, the peripubertal AO male rats were cannulated intracerebroventriculary and treated with repeated, low doses of somatostatin-14 (experimental group) or saline (control group). Twenty-four hours after the last treatment, we removed and prepared the thymuses for determination of thymocyte subpopulations by flow cytometry. After five days, animals were sacrificed and their thymuses taken for morphometrical analysis by stereological methods. We noticed that somatostatin-14 decreased volumes of thymus cortex and medulla, total number of thymocytes, number of thymocytes in the cortex and medulla and numerical density of thymocytes in deeper cortex. As a consequence of these changes, thymus size was also diminished. The phenotypic analysis of thymocyte subpopulations showed that somatostatin-14 decreased the percentage of CD4(+)CD8(+) cells with low level of TCR alphabeta expression, positively selected CD4(+)CD8(+)TCRalphabeta (high) cells and the most mature CD4(-)CD8(+)TCRalphabeta (high) cells, while the percentage of CD4(+)CD8(-)TCRalphabeta (high) thymocytes was slightly increased. Somatostatin-14 increased the relative proportion of the least mature CD4(-)CD8(-)TCRalphabeta (-/low), CD4(+)CD8(+)TCRalphabeta (-) cells and both of TCRalphabeta (-/low) single positive subpopulations. These results show that centrally applied somatostatin-14, induces hypotrophy of the thymus in peripubertal rats by changing the volumes and cellularities of the thymic compartments. Additionally, increased number of the least mature thymocytes and a deficiency of double positive cells indicate the involvement of somatostatin in the modulation of T cells maturation.

The mutation frequency of Drosophila melanogaster populations living under conditions of increased background radiation due to the Chernobyl accident.

One of the problems facing the program in the wake of the Chernobyl accident is the estimation of genetic damage to plants and animals. Special attention was directed to studying the influence of radioactive pollutants at the accident site by means of an appropriate test system, using standard genetic subjects. The present study describes such investigations. Levels of persistent genetic damage in natural populations of Drosophila melanogaster found in the vicinity of the Chernobyl accident site were examined from August 1986 to September 1989. Evidence is presented which indicates a relationship between the levels of radioactive pollution resulting from the Chernobyl accident and increasing genetic damage to exposed populations. The possible reasons for the decrease of mutation frequency observed in 1988 and 1989 are also discussed. Furthermore, evidence is presented which suggests that radiosensitive Drosophila mutants may be particularly sensitive indicators of radioactive pollution.

The established Yersinia pestis biovars are characterized by typical patterns of I-CeuI restriction fragment length polymorphism.

The genomes of three main biovars of Yersinia pestis were subjected to restriction fragment length polymorphism analysis using I-CeuI endonuclease. I-CeuI which is encoded by a mobile intron in Chlamydomonas engamenans recognizes a 25-bp site in the ribosomal RNA rrl gene and cuts DNA of most representatives of Enterobacteriaceae into seven fragments corresponding to the presence of seven rrn-operons. Glycerol-positive Y. pestis strains (biovars antiqua and mediaevalis) contain seven ribosomal operons which can be recognized by I-CeuI endonuclease. However, glycerol-negative strains of Y. pestis biovar orientalis expose only six restriction sites for I-CeuI. The restriction fragment length polymorphism patterns obtained with I-CeuI make it possible to distinguish between three biovars of Y. pestis. Use of another rare cutting restriction enzyme, Bln/I, permits differentiation between pigment-adsorbing and avirulent non-pigment-adsorbing Y. pestis. Still, due to homologous recombination between the two copies of IS 100 insertion sequence bracketing the pgm-locus, the mechanism of deletions in the pgm-locus seems to be confined only to strains of biovars antiqua and mediaevalis, and can be different in Y. pestis strains of biovar orientalis. The I-CeuI restriction patterns of two Yersinia strains isolated within a ten-year period in the port of St. Petersburg and originally identified as Y. pseudotuberculosis 01 turned out to be related to typical representatives of Y. pestis biovar antiqua. These strains could be exported from the same source or circulate among Rattus norvegicus population of the port as non-pigment-adsorbing avirulent immunogenic clone.

[Isolation of transmissible cointegrates of Yersinia pestis plasmids pYV and pYT with the plasmid RP4::Mu cts 62, IncP1]. / Poluchenie tramsmissivnykh kointegratov plazmid pYV i pYT chumnogo mikroba s plazmidoi RP4::Mu cts62, IncP1.

The transmissible cointegrates of the Yersinia pestis plasmids pYV and pYT with the broad host range plasmid RP4::Mu cts62 of the incompatibility group IncP have been constructed by the in vivo recombination. The cointegrative plasmid pKR14 (pYV76 omega RP4::Mu cts62) conferred on the transconjugants the properties of Ca2(+)-dependence at 37 degrees C, V-antigen synthesis, RP4 plasmid markers (ApR, KmR, TcR), immunity to the lysis by the bacteriophage Mu cts62 and incompatibility with the homologous replicon pYV76. Cointegrates pKR103 and pKR106 (pYT omega RP4::Mu cts62) conferred on the transconjugant clones the ability to synthesize the "mouse" toxin and fraction I. The capability of Escherichia coli cells to synthesize the latter products has been demonstrated together with the deficiency of these cells to transport the synthesized fraction I to the cell surface.

[The use of mini-Mu-replicons for cloning of various structural genes of Yersinia pestis]. / Ispol'zovanie mino-Mu-replikonov dlia klonirovaniia nekotorykh strukturnykh genov chumnogo mikroba.

The possibility to clone the structural leu gene of Yersinia pestis in vivo using the mini-Mu bacteriophages with the inserted plasmid replicones has been demonstrated. The E. coli K12 transductants having obtained the Leu(+)-marker within the cloned 4.8-21 kb fragments stably inherited the leu-gene within the autonomous mini-Mu replicones. The possibility to clone other Yersinia pestis genes by the same technique has been demonstrated.

[Interaction of Yersinia pestis bacteria with bacteriophage mu]. / Vzaimodeistvie bakterii Yersinia pestis s bakteriofagom mu.

Yersinia pestis cells are shown to be sensitive to bacteriophage Mu cts62 infection. Lysis of bacteria has been shown to be more efficient on solid nutrient medium than in LB broth. 10(-5) pfu per ml is the maximal concentration of bacteriophage particles yielded from the broth cultures of bacteria. Moi 0.1 has been used to obtain such yields of bacteriophage. Lysogenization of Yersinia pestis cells has not been achieved when the standard methods of bacteriophage infection have been used. It was accomplished by the conjugal transfer of plasmid RP4::MU cts62 to Yersinia pestis from Escherichia coli. The deficiency of Yersinia pestis in producing bacteriophage Mu cts62 mature particles during the lytic cycle of bacteriophage is discussed.

[The dynamics of the frequency of cytogenetic disorders in micropopulations of murine rodents inhabiting the region of the accident at the Chernobyl Atomic Electric Power Station]. / Dinamika chastoty tsitogeneticheskikh narushenii v mikropopuliatsiiakh myshevidnykh gryzunov, obitaiushchikh v raione avarii na Chernobyl'skoi AES.

Cytogenetic examination of some species of Muridae from Chernobyl region was carried out in 1986-1993. It was revealed that the dynamic of cytogenetic aberrations in cells of animals from territories with distinct level of radioactive contamination differed.

[Results of the studies on radiation ecology and radiation biology at the Institute of Biology of Komi Science Center of Ural division of Russian Academy of Science (on 40th anniversary of the Department of Radiation Ecology)]. / Itogi issledovanii po radioékologii i radiobiologii v Institute Biologii Komi Nauchnogo tsentra Ural'skogo otdeleniia Rossiiskoi akademii nauk (k 40-letiiu otdela radioékologii).

Information about the foundation and history of the Radiation Ecology Department and results of the researches on the effect of increased background radiation level on plant and animal populations, migration of radionuclides in natural biocoenosies with increased radiation level are presented.