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A Gram-positive, facultatively anaerobic, catalase-negative, fructose-dependent strain (W13T) was isolated from the gut of honeybee (Apis mellifera). Phylogenetic analysis based on 16S rRNA gene sequencing indicated that strain W13T represents a distinct line of descent within the genus Fructobacillus , with the closest neighbours being Fructobacillus broussonetiae BCRC 81240T (98.9 % sequence similarity) and Fructobacillus durionis DSM 19113T (96.8 % sequence similarity). Comparative sequencing of the additional phylogenetic markers rpoC and recA confirmed the 16S rRNA gene tree topology. The complete genome of strain W13T consisted of 1 292 712 bp with a G+C content of 48.3 mol%. Pairwise comparisons of the average nucleotide identity values and digital DNA–DNA hybridization values between the genomes of W13T and its close phylogenetic neighbours, F. broussonetiae BCRC 81240T and F. durionis DSM 19113T, resulted in 76.2–84.1 % and 20.2–27.6 %, respectively. The main cellular fatty acids of strain W13T were C16 : 0, C18 : 1 ω9c and C18 : 1 ω7c. Thus, we propose a novel species within the genus Fructobacillus , with the name Fructobacillus apis sp. nov. and the type strain is W13T (= NBRC 115637T=BCRC 81365T).

Sequences targeted at the V3 and V4 16S rRNA hypervariable regions of a streptococcal strain (P1L01T) isolated from vaginal swabs of a pregnant woman with diabetes were 100% similar to those of Streptococcus anginosus subsp. whileyi. However, phylogenetic analysis based on 16S rRNA full-gene sequencing (1562 bp) revealed highest sequence similarity to Streptococcus periodonticum (98.7%), followed by Streptococcus anginosus subsp. whileyi (98.7%), and Streptococcus anginosus subsp. anginosus (98.4%). Phylogenies of housekeeping genes rpoB and groEL were compared to improve classification, and the results showed a clear separation between strain P1L01T and closely related Streptococcus type strains. The complete genome of strain P1L01T consisted of 2,108,769 bp with a G + C content of 38.5 mol%. Average nucleotide identity values, based on genome sequencing, between strain P1L01T and Streptococcus periodonticum KCOM 2412T, Streptococcus anginosus subsp. whileyi CCUG 39159T, and Streptococcus anginosus subsp. anginosus NCTC 10713T were 95.5%, 94.3%, and 95.3%, respectively. The highest in silico DNA–DNA hybridization value with respect to the closest species was 66.2%, i.e., below the species cutoff of 70% hybridization. The main cellular fatty acids of strain P1L01T were 16:0, 18:1ω7c, and 14:0. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify this isolate as representative of a novel species of the genus Streptococcus, Streptococcus vaginalis sp. nov., in reference to its isolation from vaginal swabs, with strain P1L01T (= NBRC 114754T = BCRC 81289T) as the type strain.

A new α-haemolytic streptococcal strain, designated DM3B3

A novel lactic acid bacterium, strain MB7T, was isolated from lychee in Taiwan. MB7T is Gram-staining-positive, catalase-negative, non-motile, non-haemolytic, facultatively anaerobic, coccoid-shaped, heterofermentative and mainly produces d-lactic acid from glucose. Comparative analysis of 16S rRNA, pheS and rpoA gene sequences has demonstrated that the novel strain represented a member of the genus Leuconostoc . 16S rRNA gene sequencing results indicated that MB7T had the same sequence similarity of 99.25 % to four type strains of members of the genus Leuconostoc : Leuconostoc mesenteroides subsp. dextranicum DSM 20484T, Leuconostoc mesenteroides subsp. jonggajibkimchii DRC 1506T, Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293T and Leuconostoc suionicum DSM 20241T. Additionally, high 16S rRNA sequence similarities were also observed with Leuconostoc mesenteroides subsp. cremoris ATCC 19254T (99.12 %) and Leuconostoc pseudomesenteroides NRIC 1777T (98.69 %). When comparing the genomes of these type strains, the average nucleotide identity values and digital DNA–DNA hybridization values of MB7T with these type strains were 76.57–80.53 and 22.0–22.6 %, respectively. MB7T also showed different phenotypic characteristics to other most closely related species of the genus Leuconostoc , such as carbohydrate metabolizing ability, halotolerance and growth at various pHs. On the basis of phenotypic and genotypic properties, strain MB7T represents a novel species belonging to the genus Leuconostoc , for which the name Leuconostoc litchii sp. nov. is proposed. The type strain is MB7T (=BCRC 81077T=NBRC 113542T).

A coccus-shaped organism, designated ALS3T, was isolated from fresh coffee cherries collected at a farm located in the Ali Mountain region of Taiwan. Sequence analysis of its 16S rRNA gene indicated that strain ALS3T belongs to the genus Enterococcus and has more than 98.5 % sequence similarity to Enterococcus pallens and Enterococcus hermanniensis . When comparing the ALS3T genome with these two type strains, the average nucleotide identity values and digital DNA–DNA hybridization values were 72.6–73.3 and 19.2 %, respectively. The G+C content of the genomic DNA from strain ALS3T was 35.6 mol%. Results of sequence analysis, together with enzymatic activities and characteristics of carbohydrate metabolism, indicated that strain ALS3T is distinct and represents a novel species, for which the name Enterococcus alishanensis sp. nov. is proposed. The type strain is ALS3T (=NBRC 109593T=BCRC 80605T).

Sequences targeted at the V3 and V4 16S rRNA hypervariable regions of a streptococcal strain (P1L01T) isolated from vaginal swabs of a pregnant woman with diabetes were 100% similar to those of Streptococcus anginosus subsp. whileyi. However, phylogenetic analysis based on 16S rRNA full-gene sequencing (1562 bp) revealed highest sequence similarity to Streptococcus periodonticum (98.65%), followed by Streptococcus anginosus subsp. whileyi (98.65 %), and Streptococcus anginosus subsp. anginosus (98.44%). Phylogenies of housekeeping genes rpoB and groEL were compared to improve classification, and the results showed a clear separation between strain P1L01T and closely related Streptococcus type strains. The complete genome of strain P1L01T consisted of 2,108,769 bp with a G+C content of 38.5 mol%. Average nucleotide identity values, based on genome sequencing, between strain P1L01T and Streptococcus periodonticum KCOM 2412T, Streptococcus anginosus subsp. whileyi CCUG 39159T, and Streptococcus anginosus subsp. anginosus NCTC 10713T were 95.48%, 94.33%, and 95.28%, respectively. The highest in silico DNA-DNA hybridization value with respect to the closest species was 66.2%, i.e., below the species cut-off of 70% hybridization. The main cellular fatty acids of strain P1L01T were 16:0, 18:1ω7c, and 14:0. On the basis of phylogenetic, genotypic and phenotypic data, we propose to classify this isolate as representative of a novel species of the genus Streptococcus, Streptococcus vaginalis sp. nov., in reference to its isolation from vaginal swabs, with strain P1L01T (=NBRC 114754T = BCRC 81289T) as the type strain.

Leuconostoc pseudomesenteroides 607, isolated from persimmon fruit, was found to have high inhibitory activity against Listeria monocytogenes and several other Gram-positive bacteria. Inhibitory substances were purified from culture supernatant by ion-exchange chromatography, Sep-Pak C18 cartridge, and reverse-phase high-performance liquid chromatography (RP-HPLC). Two antibacterial peptides were observed during the purification procedures. One of these peptides had a molecular size of 4623.05 Da and a partial N-terminal amino acid sequence of NH2-KNYGNGVHxTKKGxS, in which the YGNGV motif is specific for class IIa bacteriocins. A BLAST search revealed that this bacteriocin was similar to leucocin C from Leuconostoc mesenteroides. Leucocin C-specific primers were designed and a single PCR product was amplified. Analysis of the nucleotide sequence has revealed a putative peptide differing by only one amino acid residue from the sequence of leucocin C. No identical peptide or protein has been reported in the literature, and this peptide, termed leucocin C-607, was therefore considered to be a new variant of leucocin C produced by Leuc. pseudomesenteroides 607. Another antibacterial peptide purified from the same culture supernatant had a molecular size of 3007.7 or 3121.97 Da. However, detailed information regarding this second peptide remains to be determined. Distinct characteristics, such as heat stability and inhibitory spectrum, were observed for the two bacteriocins produced by Leuc. pseudomesenteroides 607. These results suggested that Leuc. pseudomesenteroides 607 produces leucocin C-607 along with another unknown bacteriocin.

The bioflavonoid apigenin has been shown to possess cancer-preventive and anti-cancer activities. In a drug screening, we found that apigenin can inhibit Wnt/β-catenin signaling, a pathway that participates in pivotal biological functions, which dis-regulation results in various human diseases including cancers. However, the underlying mechanism of apigenin in this pathway and its link to anti-cancer activities remain largely unknown. Here we showed that apigenin reduced the amount of total, cytoplasmic, and nuclear β-catenin, leading to the suppression in the β-catenin/TCF-mediated transcriptional activity, the expression of Wnt target genes, and cell proliferation of Wnt-stimulated P19 cells and Wnt-driven colorectal cancer cells. Western blotting and immunofluorescent staining analyses further revealed that apigenin could induce autophagy-mediated down-regulation of β-catenin in treated cells. Treatment with autophagy inhibitors wortmannin and chloroquine compromised this effect, substantiating the involvement of autophagy-lysosomal system on the degradation of β-catenin during Wnt signaling through inhibition of the AKT/mTOR signaling pathway. Our data not only pointed out a route for the inhibition of canonical Wnt signaling through the induction of autophagy-lysosomal degradation of key player β-catenin, but also suggested that apigenin or other treatments which can initiate this degradation event are potentially used for the therapy of Wnt-related diseases including cancers.

Weissella cibaria 110 was isolated from plaa-som, a Thai fermented fish product, and known to produce the weissellicin 110 bacteriocin. We carried out comprehensive comparative genomic analysis of W. cibaria 110 with four other non-bacteriocin-producing W. cibaria strains and identified potential antibiotic-resistant genes. We further identified a type III restriction-modification system, a TA system, and a bacteriocin gene cluster that are unique in W. cibaria 110. Genes related to bacteriocin biosynthesis are organized in clusters and are encoded with minimum genetic machinery consisting of structural cognate immunity genes, including ABC transporter and immunity protein. Finally, we predicted W. cibaria 110 to produce a class IId bacteriocin, weissellicin 110, which is 31 amino acids in length and contains a 21-amino-acid N-terminal leader peptide. This is the first bacteriocin-producing sequencing genome in W. cibaria, and we describe the difference between the bacteriocin-producing and non bacteriocin-producing strains from genome point of view.

We evaluated the physiological and antioxidant characteristics of Arabidopsis thaliana (At) plants grown in different sea water (SW) products containing trace elements, namely RO3, 300K, and 340K, at various dilutions. The synthetic water (namely 300K-Test), a mixture of the main ions of SW including 143.08 mg L-1 Mg2+, 5.74 mg L-1 Na+, 170 mg L-1 K+, and 33.5 mg L-1 Ca2+ with equal concentrations to those in 300K SW without trace elements, was also used to culture At plants and study the influences that the major ions had on regulating ethylene production. The ethylene-biosynthesis (ACS7 and ACO2) and senescence-associated (NAP, SAG113, and WRKY6) gene expressions in SW- and ionic-treated At plants in response to transcriptional signaling pathways of ethylene response mechanisms were also investigated. Our results show that down-regulation of the ACS7 gene in 300K-treated plants significantly reduced the ethylene content but remarkably increased chlorophyll, total phenol, and DPPH radical scavenging accumulations and strengthened the salt tolerance of 300K-treated plants. The expression of the ACS7 gene of At plants under 300K, Ca2+, Mg2+, and Na+ treatments was correlated with decreases in NAP, SAG113, and WRKY6 gene expressions. The application of Ca2+ increased total phenol content and reduced the accumulation of superoxide, which in combination decreases plant aging brought on by ethylene. However, K+ treatment inhibited SGA113 gene expression, resulting in reducing ACS7 gene expression and ethylene content. The characterization and functional analysis of these genes should facilitate our understanding of ethylene response mechanisms in plants.

Two Gram-stain-positive, catalase-negative, rod-shaped, bacterial strains (313T and 311) were isolated from banana fruits in Taiwan. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the highest similarity to both strains corresponded to the type strain of Lactobacillus nantensis (99.19 %), followed by Lactobacillus crustorum (98.99 %), Lactobacillus heilongjiangensis (98.59 %) and Lactobacillus farciminis (98.52 %). Phylogenetic analysis based on the sequences of two housekeeping genes, pheS and rpoA, revealed that these two strains were well separated from the Lactobacillus reference strains. DNA–DNA relatedness values revealed genotype separation of the two strains from the above four species. The DNA G+C content of strain 313T was 35.5 mol%. The strains were homofermentative and mainly produced l-lactic acid from glucose. The major cellular fatty acids of strain 313T were 18 : 1ω6c and/or 18 : 1ω7c, 16 : 0, and 19 : 1ω6c and/or 19 : 0 cyclo ω10c. Based on their physiological and genotypic characteristics, the isolates represent a novel species of the genus Lactobacillus , for which the name Lactobacillus musae sp. nov. is proposed. The type strain is 313T=NBRC 112868T=BCRC 81020T).

Weissellicin 110 is the only bacteriocin reported inProviding a rapid detection method to isolate the weissellicin 110 encoding gene and determination of the bacteriocin distribution were the objectives.Bacteriocin fromThe molecular weight and partial protein sequence were obtained for the bacteriocin fromThe genetic information of weissellicin 110 was obtained, enabling rapid detection of the weissellicin 110 encoding gene. Results suggest that weissellicin 110 producing

Banana is a popular fruit worldwide. The lactic acid bacteria (LAB) microflora in banana fruits has not been studied in detail. A total of 164 LAB were isolated from banana fruits in Taiwan. These isolates were initially divided into nine groups (r1 to r9) using restriction fragment length polymorphism analysis and 16S ribosomal DNA sequencing. Isolates belonging to Lactobacillus plantarum group were further divided into three additional groups using multiplex PCR assay targeting the recA gene. The most common bacterial genera found in banana fruits were Lactobacillus and Weissella. The distribution of LAB indicated that, in most cases, neighboring regions shared common strains, but there were still some differences between regions. On the basis of phylogenetic analysis of 16S rRNA, rpoA, and pheS gene sequences, two strains included in the genera Lactobacillus were identified as potential novel species or subspecies. In addition, a total 36 isolates were found to have bacteriocin-producing abilities. These results suggest that various LAB are associated with banana fruits in Taiwan. This is the first report describing the distribution and varieties of LAB associated with banana fruits. In addition, one potential novel LAB species was also found in this study.

A total of 102 lactic acid bacteria (LAB) were isolated from three different coffee farms in Taiwan. These isolates were classified and identified by the restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Heterofermentative Leuconostoc, and Weissella species were the most common LAB found in two farms located at an approximate altitude of 800 m. Lactococcus lactis subsp. lactis was the most common LAB found in the remaining farm was located at an approximate altitude of 1,200 m. It is therefore suggested that the altitude and climate may affect the distribution of LAB. On the basis of phylogenetic analysis, two strains included in the genera Enterococcus were considered as two potential novel species or subspecies. In addition, a total of 34 isolates showed the antifungal activity against Aspergillus flavus. Moreover, seven Lactococcus lactis subsp. lactis strains and one Enterococcus faecalis strain were found to have bacteriocin-like inhibitory substance-producing capability. These results suggest that various LAB are associated with fresh coffee cherries in Taiwan. Some of the isolates found in this study showed potential as antifungal agents.

This study investigated the distribution of the native lactic acid bacteria (LAB) population in shao-jiou-luo (fermented zoned cerith), a popular traditional fermented food in Taiwan. A total of 129 cultures of LAB were isolated from shao-jiou-luo samples, and 14 cultures were isolated from its main raw material, zoned cerith. These isolates were divided into classes by phenotype and then into groups by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. The identification results revealed 11 bacterial species belonging to five genera in shao-jiou-luo and three species belonging to two genera in zoned cerith. The most common bacterial genera in shao-jiou-luo were heterofermentative Weissella and Leuconostoc. On the other hand, Enterococcus aquimarinus was the most common species found in the raw zoned cerith sample. In conclusion, the results suggest that heterofermentative LAB species greatly contributed to the fermentation of shao-jiou-luo. This is the first report des...

The main eicosanoids inflammatory mediators, prostaglandins and leukotrienes, are both generated from arachidonic acid (AA; 20:4 n-6). AA is a member of polyunsaturated fatty acids (PUFAs). Numerous studies have demonstrated that various contents of PUFAs can modulate the inflammatory responses. However, fewer studies have examined n-9PUFAs and their effects on the inflammatory responses. In the present study, the role of 5,8,11-cis-eicosatrienoic acid (ETrA; 20:3 n-9, also called Mead acid) in the inflammatory responses has been investigated. The anti-inflammatory activities of ETrA were examined using an in vitro macrophage system and the inhibitory effect was confirmed by western blot analysis for iNOS and COX-2 expressions. The interactions between ETrA and COX-2 protein were simulated to produce a computer modeling protein-ligand complexes and the results suggest a possible mechanism for the effects of ETrA. In this study, we described a significant inhibition of the inflammatory activities initiated by ETrA. Since ETrA is a substance presented in the tissues of young animals, we therefore anticipate that ETrA can be utilized as a natural therapeutic supplement to inhibit inflammatory activities.

A left atrial mass associated with coronary artery disease is often diagnosed as a mural thrombus rather than other possible etiologies such as benign primary cardiac tumor (myxoma, lipoma), a malignant primary cardiac tumor (sarcoma, lymphoma), or secondary involvement for extracardiac tumors. Malignant lymphoma initially presenting as intracardiac masses is very rare. Chest computed tomography with contrast enhancement and cardiac magnetic resonance may be the best methods for distinguishing primary cardiac tumors from direct extension from adjacent mediastinal structures. We report the case of a 59-year-old man with incidentally found mediastinal diffuse large B-cell lymphoma invading the left atrium, which presented with coronary artery disease and a left atrial mass. Improvement in cardiac ventricular function heart after coronary artery bypass grafting may provide the patient with a better chance of receiving an adequate dose of chemotherapy.

Lactobacillus pobuzihii is a novel species which has been previously found in pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan. However, the lactic acid bacteria (LAB) microflora in pobuzihi has not been studied in detail. In this study, LAB from pobuzihi were isolated, identified, and characterized. A total of 196 LAB were isolated; 79 cultures were isolated from the sample collected from a manufacturing factory, 38 from pobuzihi samples collected from 4 different markets, and 79 from 2 fresh cummingcordia samples. These isolates were characterized phenotypically and then divided into eight groups (A to H) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Lactobacillus plantarum was the most abundant LAB found in most samples during the fermentation of pobuzihi. On the other hand, Enterococcus casseliflavus and Weissella cibaria were, respectively, the major species found in the two fresh cummingcordia samples. A potential novel species or subspecies of lactococcal strain was found. In addition, seven L. plantarum and five W. cibaria strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157(T). This is the first report describing the distribution and varieties of LAB existing in the pobuzihi during its fermentation process and the final product on the market.

BACKGROUND: Jiang-gua (fermented cucumbers) is a popular traditional fermented food in Taiwan. The microflora of lactic acid bacteria (LAB) in jiang-gua have not been investigated in detail. In this study, LAB from jiang-gua were isolated, characterised and identified. RESULTS: A total of 103 LAB were isolated; 70 cultures were isolated from jiang-gua samples and 33 cultures were isolated from its raw substrate, cucumber. These isolates were mainly characterised phenotypically and then divided into seven groups (A-G) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. The isolates were identified as Enterococcus casseliflavus, Leuconostoc lactis, Leuconostoc mesenteroides, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paraplantarum, Lactococcus lactis subsp. lactis, Weissella cibaria and Weissella hellenica. The antibacterial activities of the isolates were determined and 11 Lc. lactis subsp. lactis strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157T. CONCLUSION: Heterofermentative W. cibaria and Leu. lactis were the major LAB found in jiang-gua samples without soy sauce. In soy sauce-added samples, homofermentative L. pentosus and L. plantarum were the most abundant LAB. In addition, the results also suggested that HhaI and RsaI restriction enzymes could be applied to distinguish W. hellenica and Weissella paramesenteroides. Copyright © 2012 Society of Chemical Industry

BACKGROUND:Sian-sianzih (fermented clams) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in sian-sianzih have not been studied in detail. In this study, LAB from sian-sianzih were isolated, characterized and identified. RESULTS: A total of 186 cultures of LAB were isolated from seven sian-sianzih samples and 29 cultures were isolated from its main raw substrate: clams. The identification results revealed up to 11 distinct bacterial species belonging to five genera in sian-sianzih, and three species belonging to two genera in clams. The most common bacterial genera in sian-sianzih were Lactobacillus and Weissella, followed by Leuconostoc, Pediococcus and Lactococcus. A regional similarity in LAB, with differences in diversity, was observed in the current study. On the other hand, Lactococcus lactis subsp. lactis was the most common species found in raw clam samples. The results also suggested that greater LAB diversity could be observed in wild clams than in cultured ones. Furthermore, antibacterial activities of the isolates were determined, and one Weisella hellenica strain showed inhibitory activity against the indicator strain Lactobacilluas sakei JCM 1157T. A sensory assessment of seven sian-sianzih samples was also performed and the results indicated that diversity of LAB has a great effect on its aroma and taste formation. CONCLUSION: The results demonstrate that various LAB species are distributed in sian-sianzih and have a great effect on the flavor of sian-sianzih. Copyright © 2011 Society of Chemical Industry

BACKGROUND: In this study, lactic acid bacteria (LAB) were isolated, characterized and identified from jiang-sun (fermented bamboo shoots; a traditional fermented food in Taiwan). Samples were collected at seven time intervals from a fixed fermenting bucket during the fermentation process of jiang-sun and its initial ingredients, dochi (fermented soybeans) and bamboo shoots. RESULTS: A total of 57 LAB cultures were isolated; 42 cultures were isolated from jiang-sun samples and 15 cultures were isolated from dochi and bamboo shoots. These isolates were characterized phenotypically and then divided into three groups (A-C) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Alteration of microbial populations during the fermentation process was observed. While various LAB were found in the dochi and bamboo shoots, it was mostly replaced by Lactobacillus plantarum after 1 day of fermentation. Furthermore, the antibacterial activities of the isolates were determined, and one Enterococcus faecium strain showed inhibitory activity against all the indicator strains. CONCLUSION: Results suggest that L. plantarum is the main LAB present during the fermentation of jiang-sun. To our knowledge, this is the first report describing the distribution and varieties of LAB that exist in the jiang-sun fermentation process.

BackgroundHistologic chorioamnionitis (HCA) is associated with preterm delivery and with neonatal morbidity and mortality. Because HCA is usually subclinical, histologic examination of the placenta is essential for confirmatory diagnosis. In the present study, the correlations between subclinical HCA and relevant clinical and laboratory parameters were analyzed.MethodsThis was a retrospective study. We reviewed the placental histopathologic findings and the charts of patients who were admitted to our neonatal intensive care unit after delivery and their mothers between January 2007 and March 2008. A total of 77 preterm infants [gastational age (GA): 32.2 ± 3.4 weeks, birth weight (BW): 1,718 ± 554 g] were categorized as group A with histologic evidence of placental inflammation (n = 27) or group B without histologic evidence of placental inflammation (n = 50). Placental histology was studied to identify the presence of inflammatory states such as chorioamnionitis, funisitis and deciduitis. Laboratory parameters including complete blood count, differential count, and C-reactive protein (CRP) level of mothers and initial arterial blood gas, glucose level and mean blood pressure of the infants were documented. Gestational age, Apgar score, history of prolonged premature rupture of membrane (prolonged PROM), gestational diabetes mellitus, meconium-stained amniotic fluid, pregnancy-induced hypertension and signs of pre-eclampsia were also collected as clinical parameters. All data were analyzed using independent t tests and Fisher's exact test, as appropriate.ResultsGroup A newborns had a significantly lower gestational age (30.8 ± 4.1 weeks vs. 33.0 ± 2.6 weeks, p < 0.05) and higher CRP level (0.56 ± 0.92 mg/dL vs. 0.12 ± 0.14 mg/dL, p < 0.05), together with higher maternal WBC count (13,002 ± 4,344/μL vs. 10,850 ± 3,722/μL, p < 0.05) and higher rate of prolonged PROM [14/27 (51.85%) vs. 8/37 (21.62%), p < 0.05] compared with group B newborns.ConclusionWe found that HCA was significantly correlated with lower gestational age, higher CRP level of preterm infants, higher maternal WBC count, and a higher rate of prolonged PROM. Our results demonstrate a significant association between HCA with an elevated CRP level in preterm infants. These findings further confirmed the association between maternal inflammation and preterm deliveries.

Nampt/visfatin acts in both intracellular and extracellular compartments to regulate multiple biological roles, including NAD metabolism, cancer, inflammation, and senescence. However, its function in chronic inflammation and carcinogenesis in hepatocellular carcinoma (HCC) has not been well-defined. Here we use Huh-7 hepatoma cells as a model to determine how Nampt/visfatin affects cellular survival under oxidative stress. We found that the transition of Nampt/visfatin from intracellular into extracellular form was induced by H2O2treatment in 293T cells and confirmed that this phenomenon was not due to cell death but through the secretion of Nampt/visfatin. In addition, Nampt/visfatin suppressed cell viability in oxidative treatment in Huh-7 cells and acted on the inhibition of hepatoma cell growth. Oxidative stress also reduced the Nampt-mediated activation of NF-κB gene expression. In this study, we identify a novel feature of Nampt/visfatin which functions as an adipokine that can be secreted upon cellular stress. Our results provide an example to understand how adipokine interacts with chemotherapeutic treatment by oxidative stress in HCC.

Respiratory enzyme synthesis in enterobacteria is controlled in response to electron acceptor availability. The iron-sulphur protein Fnr and the sensor-regulator proteins ArcB-ArcA control respiratory gene transcription in response to oxygen and quinone pool redox status respectively. The sensor-regulator proteins NarX-NarL and NarQ-NarP control anaerobic respiratory gene expression in response to nitrate and nitrite. Our laboratory recently engineered the lac operon to replace the primary operator O1-lac with the NarL and NarP protein binding site from the nirB operon. Expression of the lacZ gene from this construct is repressed by nitrate in Nar+ strains. Here, we found that lacZ gene expression was repressed in aerated cultures of narQ+narX null strains. This repression was not observed in narX+narQ+ or narX+narQ null strains. Thus, the NarQ sensor responds to aeration as well as to nitrate and nitrite. The NarX and NarQ sensors are composed of three distinct modules: an amino-terminal sensory module, a carboxyl-terminal transmitter module and a central module of unknown function. Experiments with NarX-NarQ hybrid proteins suggest that the NarQ protein central module is necessary for response to aeration. The physiological significance of this additional sensory role for the NarQ sensor remains obscure.

The advantages of using a new automated system, the Hydra-Plus-One System equipped with 96 or 384 precision glass syringes and a non-contact microsolenoid dispenser, in setting up high-throughput low-volume sequencing reactions and PCR are described. Using the syringe-based dispenser, which is the Hydra-PP part of this system, wet dispenses of as small as 100 nL with CVs of less than 10% can be accomplished. The single-channel, non-contact microsolenoid dispenser part of the system can dispense samples as low as 100 nL (with CVs of less than 10%) at a speed of 58s per 96 dispenses into any plate format (SBS footprint). The advantages associated with the use of the Hydra-Plus-One System for setting up PCR and sequencing reactions are high precision at nanoliter-dispense range; speed; and minimal waste of precious and expensive samples. The single-channel dispenser eliminates the dead volume associated with aspirating from reservoirs or troughs and thereby reduces sample waste. In addition, virtually all material can be recovered from the dispenser. Finally, non-contact dispensing enables distribution of sample into wells without any in-between-dispenses washing requirements.

An automated high-throughput method that employs rolling circle amplification (RCA) to generate template for large-scale DNA sequencing has been developed using liquid handling systems equipped with precision glass syringes. A protocol was designed to perform the sequencing analysis from template preparation to thermal cycle sequencing within the same vessel, thus minimizing the amount of liquid handling and transfer. The amplified DNA was directly used for cycle sequencing with no need for any purification procedures. Total RCA reaction volumes as low as 500 nL generated sufficient templates for successful sequencing. Reducing the RCA total reaction volumes by a 40-fold factor, from a total of 20 μL to 500 nL, resulted in a significant reduction in cost, from $1.25/reaction to less than $0.04/reaction. Additionally, the volume of the sequencing reactions was reduced from a total of 20 to 10 μL, thus generating a further cost advantage. This high-throughput DNA sequencing protocol maximizes the speed and precision of processing while significantly reducing the cost of amplification.

The advantages of using 1, 96, or 384 precision glass syringes in automated high-throughput microdispensers in creating highly uniform and reproducible DNA, protein, and organic compound array filters and slides are described. Using the Hydra® Microdispenser and TangoTM Liquid Handling system, 0.1–5 ng (in 50–300 nL) PCR-amplified, human cancer-related genes and housekeeping genes were spotted onto nylon membranes and coated slides. Protein solutions of 50μg/mL to 1 mg/mL were spotted onto coated slides or onto MaxiSorpTM 96-well plates. Up to 6144 spots/membrane and up to 1000 spots/slide were printed. The size of the spots created by glass syringes was uniform and reproducible (precision variation of less than 5%) from spot to spot and membrane to membrane. Using a Tango 384 system, a total of ten 6144-spot filters can be produced in approximately 25 min, translating into a spotting speed of 2.5 min/membrane.

During transcription activation at FNR-dependent promoters where the DNA site for FNR overlaps the -35 element, a surface-exposed activating region in the upstream subunit of the FNR dimer interacts with the C-terminal domain of the RNA polymerase alpha subunit. Starting with a cloned fnr gene encoding a defective FNR derivative carrying substitutions in this activating region, we screened a library of random mutations to identify substitutions that restored FNR activity. Activity can be restored by substitutions at residues T118, E47 and K60. The locations of these residues identify three separate surface-exposed regions of FNR that can play a role in transcription activation. These three regions appear to be analogues of Activating Region 1, Activating Region 2 and Activating Region 3 of the cyclic AMP receptor protein, CRP: our results underscore the similarities between FNR and CRP.

Expression from the Escherichia coli nir promoter is co-dependent on Fnr (a transcription factor triggered by oxygen starvation) and on NarL or NarP (transcription factors triggered by nitrite and nitrate ions). Fnr binds to a single DNA site centred between basepairs 41 and 42 upstream from the nir transcript start, whereas NarL and NarP bind to a site upstream, centred between basepairs 69 and 70. A novel mechanism to account for co-dependence on Fnr and NarL/NarP is suggested from experiments in which the spacing between the DNA sites for Fnr and NarL/NarP was altered. DNA sequence elements located upstream of the NarL/NarP-binding site are the targets for two or more proteins that act to repress Fnr-dependent activation of the nir promoter. This inhibition is counteracted by NarL or NarP. The model has been corroborated by the effects of several deletions and single base substitutions in the nir promoter upstream sequences: these deletions and substitutions prevent the binding of the repressor proteins. One of these repressors has been identified as the Fis protein, that binds to a site located 135-149bp upstream of the nir transcript start: the binding of Fis is suppressed by a single base substitution at position -146. The other repressor protein(s) have yet to be identified, but appear to bind downstream of the DNA site for Fis: binding is suppressed by a single base substitution at position -99.

A coccal-shaped organism, designated 516T, was isolated from yan-tsai-shin (fermented broccoli stems), a traditional fermented food in Taiwan. 16S rRNA gene sequencing results showed that strain 516T had 98.9 % sequence similarity to that of the type strain Lactococcus garvieae NBRC 100934T. Comparison of three housekeeping genes, rpoA, rpoB and pheS, revealed that strain 516T was well separated from Lactococcus garvieae NBRC 100934T. DNA–DNA hybridization studies indicated that strain 516T had low DNA relatedness with Lactococcus garvieae NBRC 100934T (46.1 %). The DNA G+C content of strain 516T was 38.1 mol% and the major fatty acids were C16 : 0 (22.7 %), C19 : 0 cyclo ω8c (17.9 %) and summed feature 7 (29.0 %). Based on the evidence, strain 516T represents a novel species of the genus Lactococcus , for which the name Lactococcus formosensis sp. nov. is proposed. The type strain is 516T ( = NBRC 109475T = BCRC 80576T).

A coccal strain isolated from fresh broccoli was initially identified asEnterococcus saccharolyticus; however, molecular identification and phenotypic traits did not support this identification. DNA–DNA hybridization with the type strain ofE. saccharolyticus(76.4 % relatedness), DNA G+C content (35.7 mol%), phylogenetic analysis based on 16S rRNA,pheSandrpoAgene sequences, rep-PCR fingerprinting and profiles of cellular fatty acids, whole-cell proteins and enzyme activities, together with carbohydrate metabolism characteristics, indicated that this strain is distinct and represents a novel subspecies, for which the nameEnterococcus saccharolyticussubsp.taiwanensissubsp. nov. is proposed. The type strain is 812T( = NBRC 109476T = BCRC 80575T). Furthermore, we present an emended description ofEnterococcus saccharolyticusand proposal ofEnterococcus saccharolyticussubsp.saccharolyticussubsp. nov. (type strain ATCC 43076T = CCUG 27643T = CCUG 33311T = CIP 103246T = DSM 20726T = JCM 8734T = LMG 11427T = NBRC 100493T = NCIMB 702594T).

Lactobacillus pobuzihii E100301 T is a novel Lactobacillus species previously isolated from pobuzihi (fermented cummingcordia) in Taiwan. Phylogenetically, this strain is closest to Lactobacillus acidipiscis , but its phenotypic characteristics can be clearly distinguished from those of L. acidipiscis . We present the draft genome sequence of strain L. pobuzihii E100301 T .

Automating electrophoresis significantly reduces the time required for loading a large number of samples, increases the speed of electrophoresis analysis, and maximizes the resolution power (clear separation of fragments) of this technique. In addition, automation increases the precision of electrophoresis analysis. Here we demonstrate an automated, high-throughput method of loading 96 samples simultaneously onto an electrophoresis gel, using the Apogent Discoveries Tango™ system and the Invitrogen™ E-Gel® 96 system.

One coccal strain, designated 0905C15T, was isolated from fresh cummingcordia, which is the main ingredient of pobuzihi (fermented cummingcordia), a traditional fermented food in Taiwan. 16S rRNA gene sequencing results showed that strain 0905C15T had 98.22–98.82 % sequence similarity to that of the type strains of four Lactococcus lactis subspecies ( L. lactis subsp. lactis BCRC 12312T, L. lactis subsp. cremoris BCRC 12586T, L. lactis subsp. hordniae BCRC 80474T and L. lactis subsp. tructae BCRC 80475T). Comparison of two housekeeping genes, recA and rpoB, revealed that strain 0905C15T was well separated from the reference strains of the genus Lactococcus . DNA–DNA hybridization studies indicated that strain 0905C15T had low DNA relatedness to the four Lactococcus lactis subspecies (9.7–15.24 %). The DNA G+C content of strain 0905C15T was 39.6 mol %. Based on the evidence, strain 0905C15T represents a novel species of the genus Lactococcus , for which the name Lactococcus taiwanensis sp. nov. is proposed. The type strain is 0905C15T ( = NBRC 109049T = BCRC 80460T).

Aim To isolate and characterize bacteriocin-like inhibitory substance (BLIS)-producing lactic acid bacteria from the intestine of grey mullet. Methods and Results Inhibitory activity against at least one or more indicator strains was observed in one Enterococcus thailandicus, one Enterococcus faecium and two Lactococcus garvieae strains. Enterococcus faecium B3-8 and Ent. thailandicus B3-22 showed the greatest inhibitory activities against Listeria monocytogenes ATCC 19111 and were therefore further characterized. The results suggested that the inhibitory substances from the two strains showed similar characteristics with respect to sensitivity to heat and proteolytic enzymes. BLIS from Ent. thailandicus B3-22 was characterized by a broader inhibitory spectrum than that from Ent. faecium B3-8. SDS-PAGE revealed that the molecular size of partially purified BLISs from Ent. faecium B3-8 and Ent. thailandicus B3-22 was c. 5 and 3 kDa, respectively. The molecular mass of purified bacteriocin from Ent. thailandicus B3-22 was further determined to be 6319 Da by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The results indicated that BLIS from Ent. thailandicus B3-22 can effectively inhibit the growth of all tested L. garvieae strains. Conclusions The findings obtained in this study suggest the potential use of Ent. thailandicus B3-22 as a biocontrol agent against pathogenic L. garvieae in the aquaculture. Significance and Impact of the Study This is the first report describing the characteristics of BLIS from Ent. thailandicus that showed potential for use as a biocontrol agent in the aquaculture.

The antifungal activities of 85 lactic acid bacteria strains isolated from fermented wax gourd against the four fungal species, Penicillium oxalicum, Aspergillus flavus, Aspergillus sydowii, and Mucor racemosus, were determined. Inhibitory activity against at least one or more fungal species was observed with 27 Weissella cibaria and 11 Weissella paramesenteroides strains. Among these strains, W. cibaria 861006 and W. paramesenteroides 860509 showed greater inhibitory activities and were therefore selected for further analysis. The results suggested that the antifungal activities were originated from the organic acids produced by W. cibaria 861006 and W. paramesenteroides 860509. The application tests indicated that the growth of P. oxalicum could be effectively inhibited by W. cibaria 861006 for 6 days on grape surfaces. However, W. paramesenteroides 860509 could only remain its inhibition effect for 48 h. The findings obtained in this study suggest the potential use of W. cibaria 861006 as a bio-protective agent against fungi for agricultural purposes or ready-to-eat fresh fruit and vegetable products.

Jiang-gua (fermented cucumbers) is a popular traditional fermented food in Taiwan. The microflora of lactic acid bacteria (LAB) in jiang-gua have not been investigated in detail. In this study, LAB from jiang-gua were isolated, characterised and identified.A total of 103 LAB were isolated; 70 cultures were isolated from jiang-gua samples and 33 cultures were isolated from its raw substrate, cucumber. These isolates were mainly characterised phenotypically and then divided into seven groups (A-G) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. The isolates were identified as Enterococcus casseliflavus, Leuconostoc lactis, Leuconostoc mesenteroides, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paraplantarum, Lactococcus lactis subsp. lactis, Weissella cibaria and Weissella hellenica. The antibacterial activities of the isolates were determined and 11 Lc. lactis subsp. lactis strains showed inhibitory activity against the indicator strain Lactobacillus sakei JCM 1157(T) .Heterofermentative W. cibaria and Leu. lactis were the major LAB found in jiang-gua samples without soy sauce. In soy sauce-added samples, homofermentative L. pentosus and L. plantarum were the most abundant LAB. In addition, the results also suggested that HhaI and RsaI restriction enzymes could be applied to distinguish W. hellenica and Weissella paramesenteroides.

Sian-sianzih (fermented clams) is a popular traditional fermented food in Taiwan. The lactic acid bacteria (LAB) microflora in sian-sianzih have not been studied in detail. In this study, LAB from sian-sianzih were isolated, characterized and identified.A total of 186 cultures of LAB were isolated from seven sian-sianzih samples and 29 cultures were isolated from its main raw substrate: clams. The identification results revealed up to 11 distinct bacterial species belonging to five genera in sian-sianzih, and three species belonging to two genera in clams. The most common bacterial genera in sian-sianzih were Lactobacillus and Weissella, followed by Leuconostoc, Pediococcus and Lactococcus. A regional similarity in LAB, with differences in diversity, was observed in the current study. On the other hand, Lactococcus lactis subsp. lactis was the most common species found in raw clam samples. The results also suggested that greater LAB diversity could be observed in wild clams than in cultured ones. Furthermore, antibacterial activities of the isolates were determined, and one Weisella hellenica strain showed inhibitory activity against the indicator strain Lactobacilluas sakei JCM 1157(T) . A sensory assessment of seven sian-sianzih samples was also performed and the results indicated that diversity of LAB has a great effect on its aroma and taste formation.The results demonstrate that various LAB species are distributed in sian-sianzih and have a great effect on the flavor of sian-sianzih.

In this study, lactic acid bacteria (LAB) were isolated, characterized and identified from jiang-sun (fermented bamboo shoots; a traditional fermented food in Taiwan). Samples were collected at seven time intervals from a fixed fermenting bucket during the fermentation process of jiang-sun and its initial ingredients, dochi (fermented soybeans) and bamboo shoots.A total of 57 LAB cultures were isolated; 42 cultures were isolated from jiang-sun samples and 15 cultures were isolated from dochi and bamboo shoots. These isolates were characterized phenotypically and then divided into three groups (A-C) by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Alteration of microbial populations during the fermentation process was observed. While various LAB were found in the dochi and bamboo shoots, it was mostly replaced by Lactobacillus plantarum after 1 day of fermentation. Furthermore, the antibacterial activities of the isolates were determined, and one Enterococcus faecium strain showed inhibitory activity against all the indicator strains.Results suggest that L. plantarum is the main LAB present during the fermentation of jiang-sun. To our knowledge, this is the first report describing the distribution and varieties of LAB that exist in the jiang-sun fermentation process.

The objective of this study was to isolate, characterize, and identify lactic acid bacteria (LAB) from ripe mulberries collected in Taiwan. Ripe mulberry samples were collected at five mulberry farms, located in different counties of Taiwan. Eighty-eight acid-producing cultures were isolated from these samples, and isolates were divided into classes first by phenotype, then into groups by restriction fragment length polymorphism (RFLP) analysis and sequencing of 16S ribosomal DNA (rDNA). Phenotypic and biochemical characteristics led to identification of four bacterial groups (A to D). Weissella cibaria was the most abundant type of LAB distributed in four mulberry farms, and Lactobacillus plantarum was the most abundant LAB found in the remaining farm. Ten W. cibaria and one Lactococcus lactis subsp. lactis isolate produced bacteriocins against the indicator strain Lactobacillus sakei JCM 1157(T). These results suggest that various LAB are distributed in ripe mulberries and W. cibaria was the most abundant LAB found in this study.

Currently, results of gel electrophoresis are commonly documented in digital formats by image acquisition instruments. In this study, gel images tuned by a common image processing software package, Photoshop, were assessed to understand the transforming algorithms and their impacts on quantitative analysis. TotalLab 100, an electrophoresis gel image analysis software package, was applied for image quantitation and evaluation. The three most frequently used image tuning functions—adjustments of the brightness, contrast, and grayscale span (level) of images—were investigated using both data generated from a standard grayscale tablet and an actual electrophoresis gel image. The influences of these procedures were analyzed for the grayscale transformation between the input and output images. Although all three procedures differentially improved the visualization of the input image, adjusting the contrast of images disrupted the quantitative information because of its nonlinear transforming algorithm. Under certain conditions, adjusting the brightness or the level of images could preserve the quantitative information because of the linear transforming algorithms. It was found that when the minimum and maximum grayscales of a gel image were recognized, using a commercial software package to maximally stretch the level may significantly improve the quality of a gel image without jeopardizing quantitative analysis.

Esophageal laceration with intramural dissection is a rare type of injury but without perforation. It is difficult to differentiate from esophageal perforation at presentation time. We report the case of a 46-year-old man who was admitted to our hospital complaining of progressive chest pain, dysphagia, and odynophagia after swallowing a fish bone three days prior to admission. Esophagoscopy revealed a deep longitudinal laceration with pus discharge in the esophagus. Computed tomography of the chest revealed low posterior mediastinal abscess formation. Surgery was performed under the impression of esophageal perforation. The definite diagnosis was esophageal laceration with intramural dissection.

Pallister-Killian Syndrome (PKS) is a rare sporadic congenital anomaly disorder, characterized by multiple congenital anomalies, especially craniofacial dysmorphism. It is also associated with mental retardation, seizure, skin pigmentation, and visceral malformations such as congenital diaphragmatic hernia, congenital heart defect, anorectal anomalies, and genital malformation. This syndrome usually presents with tissue-limited mosaicism of supernumerary 12p isochromosome i (12p). Moreover, diagnosis of Pallister-Killian Syndrome (PKS) is difficult because the ratio of abnormal to normal karyotyping is much lower in peripheral lymphocytes than in skin fibroblasts. We report the first case in Taiwan, who has tetrasomy 12p mosaic in peripheral lymphocytes.