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beta-Catenin reportedly inhibits adipogenesis through the down-regulations of peroxisome proliferator-activated receptor (PPAR)gamma and CCAAT/enhancer binding protein (C/EBP)alpha. We report that baicalin, a natural flavonoid compound, inhibits adipogenesis by modulating beta-Catenin. During 3T3-L1 cell adipogenesis, beta-Catenin was down-regulated, but baicalin treatment maintained beta-Catenin expression. Anti-adipogenic effects of baicalin were significantly attenuated by beta-Catenin siRNA transfection. beta-Catenin siRNA rescued the reduced expressions of PPARgamma, C/EBPalpha, fatty acid binding protein 4 and lipoprotein lipase by baicalin. Furthermore, baicalin modulated members of the WNT/beta-Catenin pathway by maintaining the expressions of low-density lipoprotein receptor-related protein 6, disheveled (DVL)2 and DVL3. These findings suggest that beta-Catenin mediates the anti-adipogenic effects of baicalin.

Streptococcus pneumoniae is the main etiologic agent of pneumonia worldwide. Because the members of the viridans group streptococci share a high degree of DNA sequence homologies, phenotypic and genotypic discriminations of S. pneumoniae from the viridans group are difficult. A quantitative real-time PCR assay targeting the capsular polysaccharide biosynthesis gene (cpsA) was developed as a species-specific detection tool for S. pneumoniae. The specificity was evaluated using genomic DNAs extracted from 135 oral cocci strains. Twenty-seven S. pneumoniae strains tested positive, whereas 108 other strains including Streptococcus pseudopneumoniae, Streptococcus mitis, and Streptococcus oralis did not show a specific signal. The linear regression of standard curves indicated high correlations between the log numbers of S. pneumoniae cells and the C(T) values (R(2)=0.99). The minimal limit of detection was 32 fg of purified genomic DNA, equivalent to 14 genomes of S. pneumoniae. This new real-time PCR method may be very useful as a rapid and specific tool for detecting and quantifying S. pneumoniae.

The human rotavirus G1P[8] strain is one of the most common rotaviruses worldwide, including Korea. Six Korean G1P[8] human rotaviruses, isolated using cell culture techniques, were characterized on the basis of sequence differences in VP7, VP4, VP6, and NSP4 genes to elucidate the evolutionary relationships in the community. All strains had a long RNA electropherotype, supported by VP6 gene analysis, clearly associated with subgroup II specificity. The phylogenetic analysis of VP7 gene sequences showed that they all clustered into lineage I, as reported for G1 strains in Japan, China, Vietnam, and Thailand. In addition, phylogenetic analysis of the VP4 gene showed that they belong to two distinct lineages, P[8]-II and P[8]-III. With respect to the NSP4 gene, all strains belonged to genotype B. An understanding of the ecology and molecular evolution of rotaviruses circulating in the country is very important for the development of vaccines and vaccination strategies. This study provides new information concerning the genetic variability of the rotavirus strain G1P[8] occurring most commonly as a vaccine candidate.

The purpose of this study was to determine the relationship between insulin resistance, obesity and serum prostate-specific antigen (PSA) levels in healthy men with serum PSA level below 4 ng mL−1. The men included in the study cohort were 11 827 healthy male employees of the Korea Hydro and Nuclear Power Co., LTD who had undergone medical checkups including fasting glucose, fasting insulin and serum PSA between January 2003 and December 2008. Insulin resistance was calculated by homeostasis model assessment (HOMA [fasting glucose × fasting insulin]/22.5) and quantitative insulin sensitivity check index (QUICKI; 1/[log (fasting insulin) + log (fasting glucose)]). Age-adjusted body mass index (BMI) was significantly increased according to increasing quartile of insulin resistance as determined by HOMA and QUICKI, respectively, in analysis of variance (ANOVA) test and Duncan's multiple comparison test (P < 0.001), but age-adjusted serum PSA concentration was significantly decreased according to increasing quartile of insulin resistance as determined by HOMA and QUICKI (P < 0.001). Age, BMI, insulin resistance by HOMA or QUICKI were significantly independent variables to serum PSA level in a multivariate linear regression analysis (P < 0.001). Insulin resistance was a significant independent variable to serum PSA level along with BMI. Insulin resistance and BMI were negatively correlated with serum PSA level in healthy men. Insulin resistance was positively correlated with BMI.

Streptococcus pneumoniae, the leading etiological agent of pneumonia, shares a high degree of DNA sequence homology with the viridans group of streptococci. The clinical and pathological manifestations may present with different features, and discrimination between S. pneumoniae and its close viridans cocci relatives, such as Streptococcus mitis and Streptococcus oralis, is still quite difficult. The 445-bp sequences of the N-terminal region of rpoA from nine S. pneumoniae, seven S. mitis, ten S. oralis, and two related strains were determined and compared with their respective 16S rRNA gene sequences to establish their usefulness in phylogenetic analysis. Pairwise comparisons of rpoA sequences among the species showed higher rates of evolution with lower similarities (92.3-100%) than those of 16S rRNA genes (96.8-100%). The rpoA-based phylogeny generated deeper branches and presented improved discriminatory resolution than the 16S rRNA gene-based phylogeny. These results show that rpoA sequences represent a consistent, but more discriminating, genomic marker than 16S rRNA gene sequences for investigating the evolutionary relationships of Streptococcus at the species level. rpoA could be a useful marker for identifying and classifying S. pneumoniae, S. mitis, and S. oralis from closely related taxa.

Contrary to the increasing amount of knowledge regarding the functional roles of glycine-rich RNA-binding proteins (GRPs) in Arabidopsis thaliana in stress responses, the physiological functions of GRPs in rice (Oryza sativa) currently remain largely unknown. In this study, the functional roles of six OsGRPs from rice on the growth of E. coli and plants under cold or freezing stress conditions have been evaluated. Among the six OsGRPs investigated, OsGRP1, OsGRP4, and OsGRP6 were shown to have the ability to complement cold-sensitive BX04 E. coli mutant cells under low temperature conditions, and this complementation ability was correlated closely with their DNA- and RNA-melting abilities. Moreover, OsGRP1 and OsGRP4 rescued the growth-defect of a cold-sensitive Arabidopsis grp7 mutant plant under cold and freezing stress, and OsGRP6 conferred freezing tolerance in the grp7 mutant plant, in which the expression of AtGRP7 was suppressed and is sensitive to cold and freezing stresses. OsGRP4 and OsGRP6 complemented the defect in mRNA export from the nucleus to the cytoplasm in grp7 mutants during cold stress. Considering that AtGRP7 confers freezing tolerance in plants and harbours RNA chaperone activity during the cold adaptation process, the results of the present study provide evidence that GRPs in rice and Arabidopsis are functionally conserved, and also suggest that GRPs perform a function as RNA chaperones during the cold adaptation process in monocotyledonous plants, as well as in dicotyledonous plants.

One hundred forty-six fecal specimens collected between 2007 and 2008 from infants with acute gastroenteritis were screened for rotavirus by ELISA with VP6-specific antibody. One hundred twenty-three of the samples (84.2%) were confirmed to be positive for group A rotavirus (community-acquired, n = 90 [73.2%] and nosocomial, n = 33 [26.8%]), and were typed subsequently using RT-PCR and sequence analysis methods. Determination of G- and P-type combinations showed that G4P[6] (78.9%) was the most common strain, followed by G3P[8] (7.3%), G1P[8] (6.5%), G2P[4] (0.8%), G2P[6] (0.8%), G1P[6] (0.8%), and G9P[8] (0.8%) strains. Of the 97 G4P[6] strains, 62 (63.8%) were responsible for community-acquired cases and 35 (36.1%) were hospital-acquired cases. Phylogenetic analysis of the VP7 gene from the G4P[6] strains revealed that both the community-acquired and nosocomial strains were segregated to the human rotaviruses circulating world-wide, including the prototype vaccinal strain, ST3, which constituted a novel sublineage in lineage 1. Owing to the recent emergence of G4P[6] rotaviruses within the hospital, as well as in the community, the findings from this study are important since they provide new information concerning the community and nosocomial spread of rotaviruses. J. Med. Virol. 82:700–706, 2010. © 2010 Wiley-Liss, Inc.

The gene regulation by intrinsically curved DNA is one way for bacterial sensing of and response to environmental changes. Previously, we showed that the genetic element BNT2 upstream of the ecf ( eae -positive conserved fragment) operon in the Escherichia coli O157:H7 virulence plasmid (pO157) has characteristics typical of intrinsically curved DNA, including the presence of multi-homopolymeric adenine:thymine tracts (AT tracts) and electrophoretic anomaly at 4 °C. Here we report that a local intrinsic curvature induced by the two phased AT tracts within the unusual promoter sequence of BNT2 played a major role for its temperature-dependent promoter activity. The base substitution of the AT tract in the spacer DNA between the −35 and the unusual −10 regions of the BNT2 promoter with non-AT tract sequence reduced intrinsic curvature slightly at 4 °C, but greatly affected its transcriptional activity. This implies that such a local intrinsic curvature within the unusual promoter of BNT2 is important for thermoregulation of the ecf operon.

The apoptotic effect of fluoxetine (FLX), an antidepressant, against human epithelial ovarian cancer cell lines OVCAR-3 and SK-OV-3 was investigated in relation to the mitochondria-mediated cell death process and nuclear factor (NF)-kappaB activation. FLX-induced mitochondrial membrane permeability change and formation of reactive oxygen species, leading to cell death. FLX-induced increase in mitochondrial Bax levels, decrease in cytosolic Bid and Bcl-2 levels, loss of the mitochondrial transmembrane potential, cytochrome c release, caspase-3 activation and up-regulation of p53. Oxidant scavengers and Bay 11-7085 [an inhibitor of nuclear factor kappaB (NF-kappaB) activation] prevented the FLX-induced cell death, increase in phosphorylated inhibitory kappaB-alpha and NF-kappaB p65 levels, and binding of NF-kappaB p65 to DNA. Results from this study suggest that FLX may exhibit apoptotic effect against ovarian cancer cell lines by inducing the mitochondrial membrane permeability change, which leads to cytochrome c release and subsequent caspase-3 activation, through reactive oxygen species-dependent activation of NF-kappaB.

Histone deacetylase inhibitors and casein kinase 2 inhibitors have been shown to induce apoptosis. However, the combined effect of casein kinase 2 inhibition on the apoptotic effect of histone deacetylase inhibitor is unknown. We assessed the effect of casein kinase 2 inhibition on the apoptotic effect of trichostatin A in human epithelial carcinoma cell lines with respect to cell death signaling pathways. At concentrations that did not induce cell death, the casein kinase 2 inhibitor 4,5,6,7-tetrabromobenzotriazole inhibited activation of apoptotic proteins and changes in mitochondrial membrane permeability induced by the histone deacetylase inhibitor trichostatin A. These results suggest that casein kinase 2 inhibition may reduce trichostatin A-induced apoptosis in ovarian carcinoma cell lines by suppressing activation of apoptotic proteins and changes in mitochondrial membrane permeability, which both lead to caspase-3 activation. Casein kinase 2 inhibition, which does not induce a cytotoxic effect, may prevent histone deacetylase inhibitor-mediated apoptosis.

Introduction Dehydroepiandrosterone (DHEA) is a multifunctional steroid that is increasingly available as a supplement aimed at improving libido and well-being in postmenopausal women in the recent times. Together with its sulfate version, DHEA-sulfate (DHEAS), it is the most abundant steroid in humans. The clitoris is an important component of the female sexual response, with its increased vascular response during sexual arousal that results in erection. Aims To elucidate the direct effects of DHEA/DHEAS on the vasomotor reactivity of the rabbit clitoral cavernosum. Methods Twenty New Zealand white female rabbits weighing approximately 2.5–3 kg were used in the study. Main Outcome Measures The contractile response of clitoral cavernous smooth muscle strips in response to phenylephrine (PE; 10 −9 –10 −4 M) were observed in rabbits. Additionally, DHEA/DHEAS effects on phenylephrine-induced contraction and/or acetylcholine-induced relaxation of phenylephrine-induced contraction were measured. Results DHEA/DHEAS did not elicit any remarkable response in the resting state. However, both DHEA and DHEAS evoked dose-dependent relaxations of PE-induced contraction. The contractile responses to high potassium were significantly decreased in the DHEA/DHEAS-pretreated strips, compared with the DHEA/DHEAS-nontreated strips. Additionally, contractions by Bay K 8644 (10 −7 –10 −6 M) treatment were also significantly inhibited by DHEA/DHEAS. DHEA-induced relaxation responses were stronger than DHEAS-induced relaxation responses. Various K channel blockers, tetraethylammonium (TEA; 1 mM, 10 mM), 4-aminopyridine (10 µM) and glibenclamide (10 µM) did not affect the DHEA/DHEAS-induced relaxation on muscle strips contracted by PE. Relaxation responses by acetylcholine or sodium nitroprusside (SNP) were not changed after DHEA/DHEAS pretreatment. Conclusions DHEA/DHEAS was found to induce a relaxation response in rabbit clitoral cavernosal smooth muscle, and this is thought to be mediated by direct inhibition of a voltage-dependent calcium channel. Lee SY, Myung SC, Lee MY, Kim TH, Kim SC, Kim KD, Park SH, and Kim WY. The effects of dehydroepiandrosterone (DHEA)/DHEA-sulfate (DHEAS) on the contraction responses of the clitoral cavernous smooth muscle from female rabbits. J Sex Med 2009;6:2653–2660.

A Gram-positive, aerobic and non-motile bacterial strain, designated ON-33(T), was subjected to a study based on a polyphasic approach to determine its exact taxonomic position. Strain ON-33(T) grew optimally at pH 7.0-7.5 and 30 degrees C. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain ON-33(T) fell within the clade comprising Gordonia species, clustering with Gordonia soli CC-AB07(T), with which it exhibited 16S rRNA gene sequence similarity of 98.5 %. The chemotaxonomic properties of strain ON-33(T) were consistent with those shared by members of the genus Gordonia. The peptidoglycan type was based on meso-diaminopimelic acid and the whole-cell sugars were arabinose and galactose. The predominant menaquinone was MK-9(H(2)). The major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major fatty acids were C(16 : 0), iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c, 10-methyl C(18 : 0) and C(18 : 1)omega9c. The DNA G+C content was 66.9 mol%. Strain ON-33(T) exhibited a DNA-DNA relatedness value of 13 % to G. soli DSM 44995(T) and could be differentiated from G. soli by differences in phenotypic characteristics. On the basis of the data obtained, strain ON-33(T) is considered to represent a novel species of the genus Gordonia, for which the name Gordonia hankookensis sp. nov., is proposed. The type strain is ON-33(T) (=KCTC 19599(T)=CCUG 57507(T)).

The taxonomic position of a Gram-positive, non-motile, non-spore-forming coryneform, isolated from activated sludge and designated strain CAU 212(T), was investigated using a polyphasic approach. Cellular morphology, biochemical tests and chemotaxonomic investigations revealed that strain CAU 212(T) had the characteristics of the genus Corynebacterium. Comparative 16S rRNA gene sequence analysis showed that the organism formed a hitherto-unknown subline within the genus Corynebacterium. Sequence divergence values of more than 4.3 % from recognized Corynebacterium species, together with phenotypic differences, showed that the bacterium represents a previously unrecognized member of the genus Corynebacterium, for which the name Corynebacterium doosanense sp. nov. is proposed. The type strain is CAU 212(T) (=KCTC 19568(T)=CCUG 57284(T)).

The taxonomic position of a Gram-positive coccus, designated strain CAU 28T, isolated from activated sludge foam was determined by using a polyphasic approach. Based on its cellular morphology and the results of biochemical tests, strain CAU 28T was identified tentatively as a member of the genus Lactococcus. Comparative 16S rRNA gene sequence analysis showed that levels of similarity between strain CAU 28T and the type strains of recognized Lactococcus species ranged from 90.4 to 97.2 %. DNA-DNA hybridization studies showed that strain CAU 28T displayed less than 26.1 % relatedness to the type strains of recognized Lactococcus species. The rep-PCR fingerprints revealed that strain CAU 28T was well separated from reference Lactococcus species. The combined genotypic and phenotypic data indicate that strain CAU 28T represents a novel species of the genus Lactococcus, for which the name Lactococcus chungangensis sp. nov. is proposed. The type strain is CAU 28T (=KCTC 13185T =CCUG 55099T).

A Gram-negative, motile, rod-shaped bacterial strain, KSL-125(T), was isolated from an alkaline soil from Kwangchun, Korea, and its taxonomic position was investigated in a polyphasic study. Strain KSL-125(T) grew optimally at 30 degrees C, at pH 7.5-8.0 and in the presence of 0.5% (w/v) NaCl. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that strain KSL-125(T) joins the cluster comprising the two Sphingosinicella species at a bootstrap resampling value of 100%. The values for similarity between the 16S rRNA gene sequence of strain KSL-125(T) and those of the type strains of the two Sphingosinicella species were 98.9-99.0%. Strain KSL-125(T) contained Q-10 as the predominant ubiquinone and C(17:1)omega6c, C(16:1)omega7c and/or iso-C(15:0) 2-OH and C(18:1)omega7c as the major fatty acids. The major polar lipids were sphingoglycolipid, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine and an unidentified phospholipid. The DNA G+C content was 65.1 mol%. Strain KSL-125(T) was distinguishable from the two recognized Sphingosinicella species on the basis of differential phenotypic properties, DNA-DNA relatedness data and repetitive-sequence-based PCR genomic fingerprinting patterns. The phenotypic, phylogenetic and genetic data showed that strain KSL-125(T) represents a novel species of the genus Sphingosinicella, for which the name Sphingosinicella soli sp. nov. is proposed. The type strain is KSL-125(T) (=KCTC 12482(T) =DSM 17328(T)).

Five hundred four fecal specimens, collected between 2004 and 2006 from young children with acute diarrhea, were screened for rotavirus by ELISA with VP6-specific antibody. Of these samples, 394 (78.2%) were confirmed as group A rotavirus and they underwent G- and P typing using a combination of ELISA, RT-PCR, and sequence analysis methods. The dominant circulating G serotype was G1 (35.6%) followed by G3 (26.4%), G4 (14.7%), and G2 (11.9%). There was a low prevalence of G9 (1.0%) and of unusual G type rotavirus, in particular, G12 (0.5%) and G8 (0.3%). Of the P genotype rotavirus in circulation, P[8] (53.0%) was most common followed by P[6] (15.5%), P[4] (15.2%), and P[9] (2.3%). Determination of G- and P type combinations revealed that G1P[8] strains were most prevalent (25.4%), amid G3P[8] (16.8%), G2P[4] (6.3%), and G4P[6] (6.1%) strains. Unusual or rare combinations such as G2P[6], G2P[8], G3P[4], G2P[9], G1P[9], G3P[9], G12P[6], G1P[4], G3P[6], and G8P[8] were also found. Owing to the recent emergence of G8 and G12 rotavirus, the findings from this study are important since they provide new information concerning the local and global spread of rotavirus genotypes.

A Gram-stain-negative, non-pigmented, non-spore-forming, non-motile, strictly aerobic bacterial strain, designated CAU 1038T, was isolated from a sea sand sample in Modo, Republic of Korea, and its taxonomic position was examined using a polyphasic approach. Cells of strain CAU 1038T grew optimally at 30 °C, pH 7.5 in 2 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CAU 1038T formed a distinct lineage within the class Gammaproteobacteria as a separate deep branch, with 95.2 % or lower sequence similarity to representatives of the genera Haliea, Halioglobus and Chromatocurvus, and 92.3 % or lower with Luminiphilus, Pseudohaliea and Congregibacter. The major cellular fatty acids of strain CAU 1038T were C16 : 0, C16 : 1ω7c and C18 : 1ω7c. The polar lipid pattern of the isolate consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, an unidentified aminolipid and two unidentified lipids. The strain contained lipoquinone (Q-8) as the sole respiratory quinone. The G+C content of the genomic DNA was 65 mol%. On the basis of phenotypic and chemotaxonomic data, and phylogenetic inference, strain CAU 1038T represents a novel species of a new genus in the family Halieaceae, for which the name Marimicrobium arenosum gen. nov., sp. nov. is proposed. The type strain of the type species is CAU 1038T ( = KCTC 42300T = NBRC 110727T).

Previous studies have shown that the metabolic capability of colonic microflora may be at least as high as that of the liver or higher than that of the whole human body. Aldehyde dehydrogenase (ALDH) is an enzyme produced by these bacteria that can metabolize acetaldehyde, produce from ethanol to acetate. Lactococcus species, which is commonly used as a starter in dairy products, was recently found to possess the ALDH gene, and the activity of this enzyme was determined. In this study, the ALDH activity of Lactococcus chungangensis CAU 28(T) and 11 other type strains in the genus Lactococcus was studied. Only 5 species, 3 of dairy origin (Lactococcus lactis ssp. lactis KCTC 3769(T), Lactococcus lactis ssp. cremoris KCCM 40699(T), and Lactococcus raffinolactis DSM 20443(T)) and 2 of nondairy origin (Lactococcus fujiensis NJ317(T) and L. chungangensis CAU 28(T)), showed ALDH activity and possessed a gene encoding ALDH. All of these strains were capable of making cream cheese. Among the strains, L. chungangensis produced cream cheese that contained the highest level of ALDH and was found to reduce the level of acetaldehyde in the serum of mice. These results predict a promising role for L. chungangensis CAU28(T) to be used in cheese that can be developed as functional food.

A Gram-stain-negative, non-spore-forming, rod-shaped, motile, facultatively anaerobic bacterium, designated CAU 1059T, was isolated from a seawater sample from Jeju Island, Republic of Korea. The bacterium grew optimally at 37 °C, at pH 7.0 and in the presence of 2 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CAU 1059T belonged to the genus Roseovarius. It exhibited only 91.5–96.9 % sequence similarity to the type strains of recognized Roseovarius species. Similar to other species of the genus Roseovarius, strain CAU 1059T had ubiquinone-10 (Q-10) as the predominant ubiquinone and C16 : 0 and summed feature 8 (C18 : 1ω7c/ω6c) as the major fatty acids. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine; three unidentified phospholipids, two aminolipids, an aminophospholipid and nine other lipids were also found. The G+C content of the genomic DNA was 61.9 mol%. On the basis of the data provided, strain CAU 1059T should be classified as representing a novel species of the genus Roseovarius, for which the name Roseovarius aquimarinus sp. nov. is proposed. The type strain is CAU 1059T ( = KCTC 32014T = CCUG 64792T).

Hfq is the bacterial orthologue of the eukaryotic (L)Sm family of proteins found across all domains of life and potentially an ancient protein, but it has not been found in all phyletic lines. A careful search successfully identified a distant hfq orthologue in the cyanobacteria leaving the actinobacteria as the major phylum with no known hfq orthologue. A search for hfq in actinobacteria, using domain enhanced searching (DELTA-BLAST) with cyanobacterial hfq, identified a conserved actinobacterial specific protein as remotely homologous. Structural homology modelling using profile matching to fold libraries and ab initio 3D structure determination supports this prediction and suggests module shuffling in the evolution of the actinobacterial hfq. Our results provide the basis to explore this prediction, and exploit it, across diverse taxa with potentially important post-transcriptional regulatory effects in virulence, antibiotic production and interactions in human microbiomes. However, the role of hfq in gram positive bacteria has remained elusive and experimental verification will be challenging.

A Gram-stain-positive, spore-forming, rod-shaped, motile, strictly aerobic bacterium, designated CAU 1183T, was isolated from marine sand and its taxonomic position was investigated by using a polyphasic approach. The bacterium grew optimally at 30 °C, at pH 8.5 and in the presence of 2 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CAU 1183T formed a distinct lineage within the genus Oceanobacillus and exhibited the highest similarity to Oceanobacillus chungangensis CAU 1051T (97.6 %). The strain contained MK-7 as the predominant isoprenoid quinone and anteiso-C15 : 0 was the major cellular fatty acid. The cell-wall peptidoglycan contained meso-diaminopimelic acid. The polar lipid pattern of strain CAU 1183T consisted of diphosphatidylglycerol, phosphatidylglycerol and unidentified lipids, including two phospholipids, two glycolipids, a phosphoglycolipid and two lipids. The G+C content of the genomic DNA was 37.5 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic data, strain CAU 1183T should be assigned to a novel species in the genus Oceanobacillus, for which the name Oceanobacillus arenosus sp. nov. is proposed. The type strain is CAU 1183T ( = KCTC 33037T = CECT 8560T).

A Gram-stain-negative, strictly aerobic, non-motile, non-spore-forming, short rod-shaped and moderately halophilic bacterial strain, CAU 1105T, was isolated from soil on reclaimed tidal land in Modo, Republic of Korea, and its taxonomic position was investigated using a polyphasic approach. Strain CAU 1105T grows optimally at a temperature of 37 °C at pH 7 in the presence of 3 % (w/v) sea salt. Based on 16S rRNA gene sequence similarity analyses, the novel isolate was assigned to the genus Palleronia within the class Alphaproteobacteria and showed the highest 16S rRNA gene sequence similarity with Palleronia marisminoris B33T (95.4 %). Strain CAU 1105T contained ubiquinone-10 as the only respiratory quinone and C18 : 1ω7c as the major cellular fatty acid. The DNA G+C content of strain CAU 1105T was 64.3 mol%. On the basis of phenotypic differentiation, phylogenetic and chemotaxonomic data, strain CAU 1105T represents a novel species of the genus Palleronia, for which the name Palleronia soli sp. nov. is proposed. The type strain is CAU 1105T ( = KCTC 42298T = NBRC 110740T). An emended description of the genus Palleronia is also provided.

Many human gastrointestinal facultative anaerobic and aerobic bacteria possess alcohol dehydrogenase (ADH) activity and are therefore capable of oxidizing ethanol to acetaldehyde. However, the ADH activity of Lactococcus spp., except Lactococcus lactis ssp. lactis , has not been widely determined, though they play an important role as the starter for most cheesemaking technologies. Cheese is a functional food recognized as an aid to digestion. In the current study, the ADH activity of Lactococcus chungangensis CAU 28 T and 11 reference strains from the genus Lactococcus was determined. Only 5 strains, 3 of dairy origin, L. lactis ssp. lactis KCTC 3769 T , L. lactis ssp. cremoris KCCM 40699 T , and Lactococcus raffinolactis DSM 20443 T , and 2 of nondairy origin, Lactococcus fujiensis NJ317 T and Lactococcus chungangensis CAU 28 T KCTC 13185 T , showed ADH activity and possessed the ADH gene. All these strains were capable of making cheese, but the highest level of ADH activity was found in L. chungangensis , with 45.9nmol/min per gram in tryptic soy broth and 65.8nmol/min per gram in cream cheese. The extent that consumption of cheese, following imbibing alcohol, reduced alcohol uptake was observed by following the level of alcohol in the serum of mice. The results show a potential novel benefit of cheese as a dairy functional food.

In Vietnam, highly pathogenic avian influenza (HPAI), such as that caused by H5N1 viruses, is the most highly contagious infectious disease that has been affecting domestic poultry in recent years. Vietnam might be an evolutionary hotspot and a potential source of globally pandemic strains. However, few studies have reported viruses circulating in the south-central region of Vietnam. In the present study, 47 H5N1-positive samples were collected from both vaccinated and unvaccinated poultry farms in the South Central Coast region of Vietnam during 2013-2014, and their genetic diversity was analyzed. A common sequence motif for HPAI virus was identified at HA-cleavage sites in all samples: either RERRRKR/G (clades 2.3.2.1c and 2.3.2.1a) or REGRRKKR/G (clade 1.1.2). Phylogenetic analysis of HA genes identified three clades of HPAI H5N1: 1.1.2 (n=1), 2.3.2.1a (n=1), and 2.3.2.1c (n=45). The phylogenetic analysis indicated that these Vietnamese clades may have evolved from Chinese and Cambodian virus clades isolated in 2012-2013 but are less closely related to the clades detected from the Tyva Republic, Bulgaria, Mongolia, Japan, and Korea in 2009-2011. Detection of the coexistence of virus clades 2.3.2.1 and the very virulent 1.1.2 in the south-central regions suggests their local importance and highlights concerns regarding their spread, both northwards and southwards, as well as the potential for reassortment. The obtained data highlight the importance of regular identification of viral evolution and the development and use of region-specific vaccines.

Bacillus anthracis has four plasmid possible virulence genotypes: pXO1+/pXO2+, pXO1+/pXO2-, pXO1-/pXO2+ or pXO1-/pXO2-. Due to the lack of a specific chromosomal marker for B. anthracis, differentiation of the pXO1-/pXO2- form of B. anthracis from closely related Bacillus cereus group species is difficult. In this study, we evaluate the ability of sspE, pXO1 and pXO2 primers to discriminate individual B. anthracis and the B. cereus group genotypes using multiplex real-time PCR and melting curve analysis. Optimal conditions for successful multiplex assays have been established. Purified DNAs from 38 bacterial strains including 11 strains of B. anthracis and 18 B. cereus group strains were analyzed. Nine of the B. cereus group near-neighbor strains were shown by multilocus sequence typing to be phylogenetically proximate to the B. anthracis clade. We have demonstrated that the four plasmid genotypes of B. anthracis and B. cereus group near-neighbors were differentially and simultaneously discriminated by this assay.

The population structure of the Bacillus cereus group (52 strains of B. anthracis , B. cereus , and B. thuringiensis ) was investigated by sequencing seven gene fragments ( rpoB , gyrB , pycA , mdh , mbl , mutS , and plcR ). Most of the strains were classifiable into two large subgroups in six housekeeping gene trees but not in the plcR tree. In addition, several consistent clusters were identified, which were unrelated to species distinction. Moreover, interrelationships among these clusters were incongruent in each gene tree. The incongruence length difference test and split decomposition analyses also showed incongruences between genes, suggesting horizontal gene transfer. The plcR gene was observed to have characteristics that differed from those of the other genes in terms of phylogenetic topology and pattern of sequence diversity. Thus, we suggest that the evolutionary history of the PlcR regulon differs from those of the other chromosomal genes and that recombination of the plcR gene may be frequent. The homogeneity of B. anthracis , which is depicted as an independent lineage in phylogenetic trees, is suggested to be of recent origin or to be due to the narrow taxonomic definition of species.

Bacillus anthracis is a large, gram-positive, aerobic, spore-forming bacillus. Its endospores do not divide, have no measurable metabolism, and are resistant to drying, heat, UV light, gamma radiation, and many disinfectants. In some cases, spores can remain dormant for decades. B. anthracis causes a zoonotic disease, anthrax. It also causes acute and often lethal disease in humans, such as cutaneous, intestinal, and pulmonary anthrax. For a long time, this species has attracted attention because of its hardiness, dormancy, and thus its potential use as a biological weapon (12, 13). In October 2001, B. anthracis spores were used to attack human populations in Florida, New Jersey, New York, and Washington, D.C. (12), which heightened public awareness and concern about anthrax. B. anthracis infections are confirmed mainly by conventional microbiological methods, i.e., Gram staining, capsule staining, colony morphology, and biochemical characteristics (4, 18). However, because of its clinical importance and its implication concerning public security, suspected specimens are usually referred to public health laboratories for definitive identification, epidemiologic study, and susceptibility testing (28). Therefore, not only precise but also rapid identification of isolated Bacillus species is needed. In addition, it is also important to know whether detected or isolated B. anthracis strains contain virulence plasmids or not because the virulence of B. anthracis is related to encapsulating and toxin-encoding plasmids. Given this situation, genotype analysis would seem to be most appropriate for the precise differential identification of virulent B. anthracis. However, genotype analysis is not straightforward for several reasons. Phylogenetically, B. anthracis is considered a member of the “B. cereus group,” which also includes B. cereus, B. thuringiensis, and B. mycoides (18). Moreover, B. anthracis is genotypically differentiated from its close relatives, B. cereus and B. thuringiensis, only by the presence of toxin-encoding plasmids (19), and the genomes of these three species show high levels of similarity. For example, this group share almost identical 16S ribosomal DNA sequences (1), and for this reason were suggested to be one species based on multilocus enzyme electrophoresis (MLEE) (11). Moreover, the genome of B. anthracis has 11 rRNA operons, which show sequence polymorphisms at 10 positions (27). Analysis of other chromosomal genes such as gyrB (9, 35) and the 16S-23S ribosomal intergenic spacer (2), which are usually used for bacterial genotyping or phylogenetic analysis also failed to discriminate B. anthracis from B. cereus and B. thuringiensis. Furthermore, it seems to be even more difficult to differentiate them by plasmid gene analysis, because of plasmid transfer among the closest species. For example, genes in the plasmid of B. anthracis have been successfully expressed in other bacteria (30) and been reported in other Bacillus species (22). It is important to note that pXO2 can be lost naturally (32). Due to the natural competence of B. thuringiensis and B. cereus, the horizontal transfer of plasmids has been reported (8, 26, 35). The findings presented above show why the detection and identification of B. anthracis from clinical or environmental samples must be performed precisely and why B. anthracis-specific chromosomal markers should be developed to differentiate B. anthracis from its closest relatives (23). The rpoB gene, encoding the RNA polymerase β-subunit, has been used as a marker for bacterial identification and for phylogenetic study (5, 6, 14, 16, 17, 20, 25). Recently, the rpoB gene was used for the real-time PCR detection of B. anthracis (23); however, false-positive results were observed. According to Ellerbrok et al. (7), B. cereus and B. megaterium strains were also detected by real-time rpoB PCR and, therefore, a more reliable detection and identification method is required for B. anthracis chromosomal DNA. In the present study, partial rpoB sequences (318 bp), which are located downstream of those used for real-time PCR (23) and which contain a region related to rifampin resistance, Rif r (21, 33), were compared for the genotyping of B. anthracis, B. cereus, B. thuringiensis, B. mycoides, and B. megaterium. Subsequently, we undertook to identify five Korean isolates based on their rpoB sequences and to develop a simple multiplex PCR method that can be used for the rapid and differential detection of virulent B. anthracis.

Three hundred forty-eight fecal specimens collected from young children with acute diarrhea in Seoul, Korea between January 1998 and February 2000 were examined for G and P types. Of these, 205 samples (59%) were confirmed as group A rotavirus by ELISA for the detection of VP6 antigen. Confirmed rotavirus isolates were characterized using G serotyping ELISA and RT-PCR methodologies for G and P genotyping of the outer capsid proteins VP7 and VP4, respectively. Serotyping of the outer capsid protein, VP7, revealed G4 as the dominant circulating serotype (41%) followed by G1 (28%) and quite a high incidence of mixed infection (14%). Genotyping of the VP4 protein was carried out on 55 of the rotavirus isolates with the dominant type being P[8] (46%). Of interest were a number of unusual G and P type combinations detected in Korea for the first time, especially the P[4] genotype associated with non-G2 serotypes. There were also a number of P[6] isolates identified including one G2P[6] isolate.

Summary A polyphasic taxonomic study was undertaken to establish the taxonomic position of six representative strains isolated from activated sewage sludge foam. The organisms were found to have chemical and morphological properties consistent with their assignment to the genus Tsukamurella . DNA:DNA relatedness studies showed that five out of the six isolates formed a distinct genomic species, the remaining strain was most closely associated with this taxon. The five isolates had a unique phenotypic profile that served to distinguish them from representatives of the validly described species of Tsukamurella . The combination of the genotypic and phenotypic data indicated that the five strains should be classified as a new species in the genus Tsukamurella . The name proposed for this taxon is Tsukamurella spumae , the type strain is N1171 T (= DSM 44113 T = NCIMB 13947 T ). It was also shown that some of the reference strains were misclassified as Tsukamurella paurometabola .

16S rDNA sequence analysis and repetitive element sequence-based PCR (rep-PCR) genomic fingerprinting were evaluated on 11 type strains of the genus Yersinia and 17 recognized serotype strains of Y. pseudotuberculosis to investigate their genetic relatedness and to establish the value of techniques for the identification of Y. pseudotuberculosis. A phylogenetic tree constructed from 16S rDNA sequences showed that the type strains of Yersinia species formed distinct clusters with the exception of Y. pestis and Y. pseudotuberculosis. Moreover, Y. pestis NCTC 5923T was found to be closely related to Y. pseudotuberculosis serotypes 1b, 3, and 7. Dendrograms generated from REP-PCR, and ERIC-PCR data revealed that members of the genus Yersinia differed from each other with the degree of similarity 62% and 58%, respectively. However, the BOX-PCR results showed that Y. pestis 5923T clustered with the Y. pseudotuberculosis group with a degree of similarity 74%. According to these findings, 16S rDNA sequence analysis was unable to reliably discriminate Y. pseudotuberculosis from Y. pestis. However, REP-PCR and especially ERIC-PCR provided an effective means of differentiating between members of the taxa.

PURPOSE Escherichia coli sequence type (ST) 131, a multidrug-resistant clone causing extraintestinal infections, has rapidly become prevalent worldwide. However, the epidemiological and clinical features of pediatric infections are poorly understood. We aimed to explore the characteristics of ST131 Escherichia coli isolated from Korean children with urinary tract infections. METHODS We examined 114 uropathogenic E. coli (UPEC) isolates from children hospitalized at Chung-Ang University Hospital between 2011 and 2014. Bacterial strains were classified into STs by partial sequencing of seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, and recA). Clinical characteristics and antimicrobial susceptibility were compared between ST131 and non-ST131 UPEC isolates. RESULTS Sixteen UPEC isolates (14.0%) were extended-spectrum β-lactamase (ESBL)-producers; 50.0% of ESBL-producers were ST131 isolates. Of all the isolates tested, 13.2% (15 of 114) were classified as ST131. There were no statistically significant associations between ST131 and age, sex, or clinical characteristics, including fever, white blood cell counts in urine and serum, C-reactive protein, radiologic abnormalities, and clinical outcome. However, ST131 isolates showed significantly lower rates of susceptibility to cefazolin (26.7%), cefotaxime (40.0%), cefepime (40.0%), and ciprofloxacin (53.3%) than non-ST131 isolates (65.7%, 91.9%, 92.9%, and 87.9%, respectively; P

A Gram-negative bacterium, designated CAU 1040(T), which was isolated from marine sand obtained from Jeju Island in South Korea, was characterized as an aerobic rod-shaped organism that that was non-motile, non-spore-forming and halophilic. The bacterium grew optimally at 37 °C, at pH 8, and in the presence of 2% (w/v) NaCl. The taxonomic classification of CAU 1040(T) was investigated using a polyphasic characterization approach. While phylogenetic analysis of the 16S rRNA gene sequence revealed that CAU 1040(T) belongs to the genus Kangiella, the strain exhibited only 94.4-95.4% sequence similarity to the previously described Kangiella species. Similar to other Kangiella species, Q-8 was the predominant ubiquionone and iso-C(15:0) was the major cellular fatty acid detected in strain CAU 1040(T). The predominant polar lipids identified were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine. The G+C content of the CAU 1040(T) genome was 45.3 mol%. The phylogenetic, physiological, biochemical and chemotaxonomic data obtained in this study indicate that strain CAU 1040(T) represents a novel species of the genus Kangiella, for which the name Kangiella chungangensis sp. nov. is hereby proposed. The type strain is CAU 1040(T) (KCTC 42299(T), NBRC 110728(T)).

Ascochyta rabiei and Alternaria solani, the causal agents of Ascochyta blight of chickpea (Cicer arietinum) and early blight of potato (Solanum tuberosum), respectively, produce a set of phytotoxic compounds including solanapyrones A, B, and C. Although both the phytotoxicity of solanapyrones and their universal production among field isolates have been documented, the role of solanapyrones in pathogenicity is not well understood. Here, we report the functional characterization of the sol5 gene, which encodes a Diels-Alderase that catalyzes the final step of solanapyrone biosynthesis. Deletion of sol5 in both Ascochyta rabiei and Alternaria solani completely prevented production of solanapyrones and led to accumulation of the immediate precursor compound, prosolanapyrone II-diol, which is not toxic to plants. Deletion of sol5 did not negatively affect growth rate or spore production in vitro, and led to overexpression of the other solanapyrone biosynthesis genes, suggesting a possible feedback regulation mechanism. Phytotoxicity tests showed that solanapyrone A is highly toxic to several legume species and Arabidopsis thaliana. Despite the apparent phytotoxicity of solanapyrone A, pathogenicity tests showed that solanapyrone-minus mutants of Ascochyta rabiei and Alternaria solani were equally virulent as their corresponding wild-type progenitors, suggesting that solanapyrones are not required for pathogenicity.

Lactococcus lactis has a played a prominent role in food industry from traditional milk fermentations to industrial scale processes. Extensive studies on the biochemical, physiological, and genetic aspects of L. lactis are evident from published literature. Recently, another novel species, Lactococcus chungangensis, was isolated from activated sludge as the sixth member of the genus to be discovered. To date, no study has been conducted to explore the functional aspects of L. chungangensis to identify features similar to those in L. lactis that are relevant to the dairy industry. Hence, the present study was undertaken to identify functional genes relevant to dairy application through comparative transcriptomic analysis of L. chungangensis with L. lactis. In expression microarray data, 415 genes were upregulated and 1,500 were downregulated of the total 1,915 probes analyzed. Interesting findings from this study were the identification of functional genes such as aminohydrolase and S-adenosylmethionine in L. chungangensis, which are useful in flavor production in cheese making. Probing these genes by PCR and analyzing the sequence confirmed the presence of these genes. Phenotypic analysis of these genes was also investigated by growing the strain in different concentrations of skim milk media to confirm the ability of L. chungangensis to degrade casein in milk, which is the major precursor for flavor enhancing compounds. Other adaptive and stress-response genes such as cold shock and heat shock proteins were also revealed. All experimental investigations at the functional level suggest that L. chungangensis possesses some interesting genes which are of commercial significance, especially in cheese production.

A human parechovirus (HPeV) strain CAU10-NN was detected from a stool sample of a 2-year-old healthy female infant in South Korea using a metagenomic approach. The CAU10-NN virus was isolated using a cell culture system and its whole genome was analyzed. The RNA genome of the CAU10-NN strain consists of 7,348 nucleotides (nt), excluding a poly(A) tail. A large open reading frame of 6,540 nt that encodes a putative polyprotein precursor of 2,180 amino acids is flanked by a 5'-untranslated region (UTR) of 708 nt and 3'-UTR of 88 nt followed by a poly(A) tail. Phylogenetic analysis shows that the CAU10-NN strain belongs to HPeV1. SimPlot and Bootscan analyses reveal that the virus genome is composed of regions related to corresponding genomic regions of other HPeVs. Recombination analysis indicates that the CAU10-NN strain might be a product of more than one genomic recombination event that occurred among HPeV1, HPeV3, and HPeV4 strains.

Purpose We investigated the molecular types of uropathogenic Escherichia coli (UPEC) by using conventional phylogrouping, multilocus sequence typing (MLST), and fimH genotyping. Methods Samples of patients younger than 18 years of age were collected from the Chung-Ang University Hospital over 2 years. Conventional phylogenetic grouping for UPEC strains was performed by polymerase chain reaction (PCR). Bacterial strain sequence types (STs) were classified on the basis of the results of partial sequencing of seven housekeeping genes. In addition, we analyzed nucleotide variations in a 424-base pair fragment of fimH, a major virulence factor in UPEC. Results Sixty-four UPEC isolates were analyzed in this study. Phylogenetic grouping revealed that group B2 was the most common type (n=54, 84%). We identified 16 distinctive STs using MLST. The most common STs were ST95 (35.9%), ST73 (15.6%), ST131 (12.5%), ST69 (7.8%), and ST14 (6.3%). Fourteen fimH allele types were identified, of which 11 had been previously reported, and the remaining three were identified in this study. f1 (n=28, 45.2%) was found to be the most common allele type, followed by f6 and f9 (n=7, 11.3% each). Comparative analysis of the results from the three different molecular typing techniques revealed that both MLST and fimH typing generated more discriminatory UPEC types than did PCR-based phylogrouping. Conclusion We characterized UPEC molecular types isolated from Korean children by MLST and fimH genotyping. fimH genotyping might serve as a useful molecular test for large epidemiologic studies of UPEC isolates.

A Gram-stain-negative, non-motile, rod-shaped, strictly aerobic bacterial strain, designated CAU 1289T, was isolated from a marine sand and its taxonomic position was investigated using a polyphasic approach. It grew optimally at pH 6.5 and 30 °C and in the presence of 3 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CAU 1289T belonged to the genus Salegentibacter , exhibiting sequence similarity values of 93.6–96.3 % to members of this genus, and was related most closely to Salegentibacter mishustinae KMM 6049T (96.3 % similarity). Strain CAU 1289T contained MK-6 as the predominant menaquinone. The major fatty acid was iso-C15 : 0. The cell-wall peptidoglycan of strain CAU 1289T contained meso-diaminopimelic acid. The polar lipids were composed of phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylcholine, an unidentified phospholipid, an unidentified aminolipid, an unidentified aminophospholipid, an unidentified glycolipid and ten unidentified lipids. The DNA G+C content was 38.7 mol%. On the basis of phenotypic, chemotaxonomic and phylogenetic inference, strain CAU 1289T is considered to represent a novel species of the genus Salegentibacter , for which the name Salegentibacter chungangensis sp. nov. is proposed. The type strain is CAU 1289T (KCTC 24000T = CCUG 64793T). An emended description of the genus Salegentibacter is also proposed.

A Gram-stain-negative, non-spore-forming, motile, strictly aerobic bacterial strain, designated CAU 1294T, was isolated from a sand sample and its taxonomic position was investigated using a polyphasic approach. The strain grew optimally at pH 6.5 and 30 °C and in the presence of 2 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain CAU 1294T formed a lineage with member of the genus Sneathiella and exhibited similarity to Sneathiella glossodoripedis MKT133T (96.3 % similarity) and Sneathiella chinensis LMG 23452T (95.1 % similarity). Strain CAU 1294T contained Q-10 as the predominant respiratory quinone. C18 : 1ω7c, C16 : 0 and cyclo-C19 : 0ω8c were the major cellular fatty acids. The polar lipids were composed of phosphatidylethanolamine, phosphatidylmethylethanolamine and two unidentified phospholipids. The DNA G+C content was 56.6 mol%. On the basis of these results, strain CAU 1294T is considered to represent a novel species of the genus Sneathiella , for which the name Sneathiella chungangensis is proposed. The type strain is CAU 1294T ( = KCTC 32476T = CECT 8513T). An emended description of the genus Sneathiella is also proposed.

Viral gastroenteritis is the most common causal agent of public health problems worldwide. Noroviruses cause nonbacterial acute gastroenteritis in humans of all ages. In this study, we investigated the occurrence of norovirus infection in children with acute gastroenteritis admitted to university hospitals in South Korea. We also analyzed the genetic diversity of the viruses and identified novel recombination events among the identified viral strains. Of 502 children with acute gastroenteritis admitted to our three hospitals between January 2011 and March 2012, genotyping of human noroviruses was performed in 171 (34%) norovirus-positive samples. Of these samples, 170 (99.5%) were in genogroup II (GII), while only one (0.5%) was in genogroup I (GI). The most common GII strain was the GII.4-2006b variant (n = 96, 56.5%), followed by GII.6 (n = 23, 13.5%), GII.12 (n = 22, 12.9%), GII.3 (n = 20, 11.8%), GII.2 (n = 6, 3.5%), GII.b (n = 2, 1.2%), and GII.10 (n = 1, 0.6%). Potential recombination events (polymerase/capsid) were detected in 39 GII strains (22.9%), and the most frequent genotypes were GII.4/GII.12 (n = 12, 30.8%), GII.4/GII.6 (n = 12, 30.8%), GII.4/GII.3 (n = 8, 20.5%), GII.b/GII.3 (n = 3, 7.7%), GII.16/GII.2 (n = 2, 5.1%), GII.4/GII.2 (n = 1, 2.6%), and GII.2/GII.10 (n = 1, 2.6%). For the first time, a novel GII.2/GII.10 recombination was detected; we also identified the GII.16/GII.2 strain for the first time in South Korea. Our data provided important insights into new recombination events, which may prove valuable for predicting the emergence of circulating norovirus strains with global epidemic potential.

Respiratory infections are well-known triggers of chronic respiratory diseases. Recently, culture-independent tools have indicated that lower airway microbiota may contribute to pathophysiologic processes associated with asthma and chronic obstructive pulmonary disease (COPD). However, the relationship between upper airway microbiota and chronic respiratory diseases remains unclear. This study was undertaken to define differences of microbiota in the oropharynx of asthma and COPD patients relative to those in healthy individuals. To account for the qualitative and quantitative diversity of the 16S rRNA gene in the oropharynx, the microbiomes of 18 asthma patients, 17 COPD patients, and 12 normal individuals were assessed using a high-throughput next-generation sequencing analysis. In the 259,572 total sequence reads, alpha and beta diversity measurements and a generalized linear model revealed that the oropharynx microbiota are diverse, but no significant differences were observed between asthma and COPD patients. Pseudomonas spp. of Proteobacteria and Lactobacillus spp. of Firmicutes were highly abundant in asthma and COPD. By contrast, Streptococcus, Veillonella, Prevotella, and Neisseria of Bacteroidetes dominated in the healthy oropharynx. These findings are consistent with previous studies conducted in the lower airways and suggest that oropharyngeal airway microbiota are important for understanding the relationships between the various parts of the respiratory tract with regard to bacterial colonization and comprehensive assessment of asthma and COPD.

The probiotic potential of a Lactobacillus strain, isolated from pig faeces, was assessed as a probiotic in piglets. The strain was examined for resistance to pH 2.0, 0.5% oxgall and antibiotics, and antimicrobial activities against enteric pathogenic bacteria. The probiotic strain, L. reuteri BSA131, was administered through the feed to 25 1-month-old Landrace piglets. The piglets were divided into five groups of five piglets each and fed with different diets for 28 days. The daily consumption of L. reuteri BSA131 was assigned into two groups by the concentration of 10(6) or 10(8) freeze-dried bacteria. Fecal samples were collected before, during, and after consumption. Lactobacilli and enterobacteria cell counts were determined in the fecal samples. The liveweight gains and feed consumption of the piglets were recorded daily. This study showed that strain BSA131 enhanced liveweight gains and feed conversion rates in piglets. It also showed a significant increase in lactobacilli cell counts and decreases in enterobacterial numbers in the fecal samples. Strain BSA131 was considered to be a potential probiotic for piglets, especially after weaning.

Recombinant rotavirus VP6 expressed in transgenic tomato cells was found primarily in the intracellular fraction and had a molecular weight of 44 kDa. In a shake flask, transgenic tomato cells produced 0.33 mg recombinant VP6 l−1 after 18 days of incubation. In a high aspect rotating-wall vessel designed by NASA to simulate microgravity, the transgenic cells produced up to 0.15 mg recombinant VP6 l−1.

Twenty-one strains of Saccharomonospora azurea, Saccharomonospora cyanea, Saccharomonospora glauca, Saccharomonospora viridis, and “Saccharomonospora caesia” were examined to evaluate the discriminatory value of 16S ribosomal DNA (rDNA) fingerprints. The 16S rDNAs were amplified by PCR by using oligonucleotide primers complementary to 16S rRNA genes. A restriction fragment length polymorphism (RFLP) analysis of the 16S rDNAs was performed with SmaI and MluI. The four validly described Saccharomonospora species could be differentiated on the basis of their characteristic 16S rDNA restriction patterns. The strains of “S. caesia” gave a restriction pattern identical to that of S. azurea K161T (T = type strain). This result was anticipated from the previous report that S. azurea K161T and the strains of “S. caesia” have identical 16S rRNA sequences. We found that purification of amplified 16S rDNA products following PCR was necessary for our RFLP analysis.

A Gram-stain-positive, spore-forming, rod-shaped, motile, strictly aerobic bacterial strain, designated CAU 1033T, was isolated from soil and its taxonomic position was investigated using a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CAU 1033T formed a distinct lineage within the genus Bacillus and was most closely related to Bacillus drentensis KCTC 13025T (similarity 95.9 %). CAU 1033T contained MK-7 as the only isoprenoid quinone and iso-C15 : 0 and anteiso-C15 : 0 as the major fatty acids. The cell wall peptidoglycan of strain CAU 1033T contained meso-diaminopimelic acid and the major whole-cell sugars were arabinose, sucrose and ribose. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, two unidentified phospholipids, four unidentified aminophospholipids, an unidentified aminolipid, two unidentified glycolipids and another unidentified polar lipid. The DNA G+C content was 41.4 mol%. On the basis of phenotypic data and phylogenetic inference, strain CAU 1033T was classified as a representative of a novel species in the genus Bacillus for which the name Bacillus songklensis sp. nov. is proposed. The type strain is CAU 1033T ( = KCTC 13881T = CCUG 61889T).

A Gram-stain-positive, spore-forming, rod-shaped, motile, strictly aerobic bacterial strain, designated CAU 1051T, was isolated from a sand dune and its taxonomic position was investigated using a polyphasic approach. Strain CAU 1051T grew optimally at pH 5.0 and 30 °C. NaCl was not required for growth but up to 10.0 % (w/v) NaCl was tolerated. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CAU 1051T formed a distinct lineage within the genus Oceanobacillus and was most closely related to Oceanobacillus profundus CL-MP28T, Oceanobacillus caeni S-11T, and Oceanobacillus picturae LMG 19492T (96.8 %, 95.6 % and 95.3 % similarity, respectively). DNA–DNA reassociation analysis showed that strain CAU 1051T displayed 28.2±0.7 % relatedness to O. profundus KCTC 13625T. Strain CAU 1051T contained MK-7 as the only isoprenoid quinone and anteiso-C15 : 0 as the major fatty acid. The cell wall peptidoglycan of strain CAU 1051T contained meso-diaminopimelic acid. The polar lipids were composed of diphosphatidylglycerol, phosphatidylglycerol, six unidentified phospholipids, an unidentified glycolipid, and six unidentified polar lipids. The major whole-cell sugars were glucose and ribose. The DNA G+C content was 36.3 mol%. On the basis of phenotypic data and phylogenetic inference, strain CAU 1051T represents a novel species of the genus Oceanobacillus for which the name Oceanobacillus chungangensis sp. nov. is proposed. The type strain is CAU 1051T ( = KCTC 33035T = CCUG 63270T).

During an epidemiological survey of human rotavirus infection in Seoul, Korea, from 2010 to 2011, one isolate of group C rotavirus (GCRV), named CAU10-312, was detected in a 5-year-old child admitted to the hospital with acute gastroenteritis, and its complete genomic sequence was determined. The 11 gene segments of the strain possessed G4-P[2]-I2-R2-C2-M2-A2-N2-T2-E2-H2 genotypes. The genotype of strain CAU10-312 appears to be closely related to strains from Bangladesh (DhakaC13 and BS347), India (v508), and England (Bristol), but distinct from Far East Asian strains, Chinese (Wu82 and YNR001) and Japanese (OH567 and BK0830). These findings may clarify the relationship of the genetics, evolutionary biology, and epidemiology of GCRVs and suggest that two very distinct genotype strains are in circulation in the world.

A rare rotavirus, RVA/Human-wt/KOR/CAU12-2/2012/G11P[25], was isolated from a 16-year-old female with fever and diarrhea during the 2012 rotavirus surveillance in South Korea using a cell culture system, and its full genome sequence was determined and analyzed. Strain CAU12-2 exhibited a G11-P[25]-I12-R1-C1-M1-A1-N1-T1-E1-H1 genotype constellation. Phylogenetic analysis of this strain revealed that it is a human-porcine reassortant of two distant relatives of the G11 strains circulating in the world. The VP7 and VP4 genes are most closely related to those of human G11P[25] viruses (Dhaka6, KTM368, and N-38 strains) identified in South Asia, whereas the VP1 gene originated from a porcine G11P[7] virus (YM strain) that was identified in South America. The VP6 gene was found to belong to the new genotype I12. This study indicates that the G11-P[25]-I12 genotype was introduced into the South Korean population by interspecies transmissions of human and animal rotaviruses, followed by multiple reassortment events.

Substance P is an injury-inducible endogenous factor for the mobilization of CD29+ stromal-like cells into circulation and that are major effectors of accelerated healing. In this study, we evaluated the effect of intravenously injected substance P on full-thickness skin wound healing as a secondary intention wound model. We made circular full-thickness skin wounds on the ears of 28 New Zealand white rabbits. They were treated with phosphate-buffered saline, or intravenous 5, 50, or 250 n mole/kg substance P at days 0 and 1. All substance P-treated groups showed a 2.6-5.4-fold higher CD29 expression and resulted in greatly decreased wound contraction and early maturation of the stroma. However, a significant decrease in wound contraction was measured only in the 5 n mole/kg treatment group. We conclude that intravenously injected substance P at 5 n mole/kg decreases wound contraction and promotes wound maturation in full-thickness skin wounds in a rabbit ear model.