Incident fracture associated with increased risk of mortality even after adjusting for frailty status in elderly Japanese men: the Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) Cohort Study.

Frail elderly individuals have elevated risks of both fracture and mortality. We found that incident fractures were associated with an increased risk of death even after adjusting for pre-fracture frailty status as represented by physical performance tests and laboratory tests for common geriatric diseases in community-dwelling elderly Japanese men. INTRODUCTION: While fractures reportedly increase the risk of mortality, frailty may complicate this association, generating a false-positive result. We evaluated this association after adjusting for pre-fracture levels of frailty. METHODS: We examined 1998 community-dwelling ambulatory men aged ≥65 years at baseline in the Fujiwara-kyo Osteoporosis Risk in Men Study for frailty status as represented by activities of daily living (ADL), physical performance tests (grip strength, one-foot standing balance with eyes open, timed 10-m walk), and laboratory sera tests. Participants were then followed for 5 years for incident clinical fractures and death. Effects of incident fracture on death were determined by Cox proportional hazards model with the first fracture during follow-up as a time-dependent predictor and with frailty status indices as covariates. RESULTS: We identified 111 fractures in 99 men and 138 deaths during the follow-up period (median follow-up, 4.5 years). Participants with incident fractures did not have significantly worse frailty statuses, but did show a significantly higher cumulative mortality rate than those without fractures (p = 0.0047). Age-adjusted hazard ratio (HR) of death for incident fracture was 3.57 (95 % confidence interval: 2.05, 6.24). When adjusted for physical performance, this decreased to 2.77 (1.51, 5.06), but remained significant. The HR showed no significant change when adjusted for laboratory test results (3.96 (2.26, 6.94)). Exclusion of deaths within the first 24 months of follow-up did not alter these results. CONCLUSION: Incident clinical fracture was associated with an elevated risk of death independently of pre-fracture levels of frailty in community-dwelling elderly men.

Isolation and characterization of human intestinal Enterococcus avium EFEL009 converting rutin to quercetin.

UNLABELLED: Quercetin is a flavonol believed to have beneficial effects on human health. Rutin, found in many plants, fruits and vegetables, is metabolized by human intestinal bacteria and converted to quercetin, where it is absorbed through the intestinal epithelium. This study aimed to isolate and characterize human intestinal bacteria capable of converting rutin to quercetin. A bacterium that can metabolize rutin was isolated from human faecal samples and identified by 16S rRNA gene sequencing. The whole-cell enzymatic activities on flavonoid glycoside and the conversion profiles of the isolate were also analysed. The bacterium was identified as Enterococcus avium EFEL009 and was shown to convert rutin to isoquercetin and then to quercetin under anaerobic conditions. Microscopic analysis revealed short chains of cocci with diameters of approx. 1 µm. ß-Glucosidase was shown to be constitutively expressed in Ent. avium, while α-rhamnosidase was expressed following induction by rutin. Both enzymes were mainly localized to the cell surface. This study is the first report on the isolation of a quercetin-producing Ent. avium FEEL009, which could be a potential industrial starter bacterium. SIGNIFICANCE AND IMPACT OF THE STUDY: Quercetin is a member of the flavonoids family reported to have better cytoprotective abilities, stronger inhibition of lipopolysaccharide-induced nitric oxide production, and better chemoprevention than rutin. This is the first report on the isolation and characterization of Enterococcus avium EFEL009 from the human intestine which is capable of converting rutin to quercetin.

Trabecular bone score may improve FRAX® prediction accuracy for major osteoporotic fractures in elderly Japanese men: the Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) Cohort Study.

UNLABELLED: FRAX® is widely used to evaluate fracture risk of individuals in clinical settings. However, FRAX® prediction accuracy is not sufficient, and improvement is desired. Trabecular bone score, a bone microarchitecture index, may improve FRAX® prediction accuracy for major osteoporotic fractures in community-dwelling elderly Japanese men. INTRODUCTION: To improve fracture risk assessment in clinical settings, we evaluated whether the combination of FRAX® and Trabecular Bone Score (TBS) improves the prediction accuracy of major osteoporotic fractures (MOFs) in elderly Japanese men compared to FRAX® alone. METHODS: Two thousand and twelve community-dwelling men aged ≥65 years completed the Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) Baseline Study comprising lumbar spine (LS) and femoral neck areal bone mineral density (aBMD) measurements, and interviews regarding clinical risk factors required to estimate 10-year risk of MOF (hip, spine, distal forearm, and proximal humerus) using the Japanese version of FRAX® (v.3.8). TBS was calculated for the same vertebrae used for LS-aBMD with TBS iNsight software (v.2.1). MOFs that occurred during the follow-up period were identified by interviews or mail and telephone surveys. Prediction accuracy of a logistic model combining FRAX® score and TBS compared to FRAX® alone was evaluated by area under receiver-operating characteristic curves (AUCs), as well as category-free integrated discrimination improvement (IDI) and net reclassification improvement (NRI). RESULTS: We identified 22 men with MOFs during 8140 person-years (PY) of follow-up among 1872 men; 67 men who suffered from fractures other than MOFs were excluded. Participants with MOFs had significantly lower TBS (p = 0.0015) and higher FRAX® scores (p = 0.0089) than those without. IDI and NRI showed significant improvements in reclassification accuracy using FRAX® plus TBS compared to FRAX® alone (IDI 0.006 (p = 0.0362), NRI 0.452 (p = 0.0351)), although no difference was observed in AUCs between the two. CONCLUSIONS: TBS may improve MOF prediction accuracy of FRAX® for community-dwelling elderly Japanese men.

Greater milk intake is associated with lower bone turnover, higher bone density, and higher bone microarchitecture index in a population of elderly Japanese men with relatively low dietary calcium intake: Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) Study.

UNLABELLED: The effects of milk intake on bone health are not clear in elderly Asian men with low dietary calcium intake. This study showed that greater milk intake is associated with lower bone turnover, higher bone density, and higher bone microarchitecture index in community-dwelling elderly Japanese men. INTRODUCTION: The consumption of milk or dairy products is widely recommended for maintaining bone health regardless of gender or age. However, little evidence exists on the beneficial effects of milk intake on bone health in elderly Japanese men characterized with relatively low dietary calcium intake. Here we examined whether or not greater milk intake was associated with lower bone turnover, higher bone density, and stronger bone microarchitecture in community-dwelling elderly Japanese men. METHODS: Interviews were conducted to obtain information on medical history and lifestyle, including the amount of habitual milk intake, nutrient intake calculations based on a 1-week food diary, and measurements of areal bone mineral density (aBMD) at the lumbar spine (LS), total hip (TH), and femoral neck (FN) by dual-energy x-ray absorptiometry (DXA), trabecular bone score (TBS) using DXA images at LS, and biochemical markers of bone turnover in sera. Participants with a history of diseases or medications that affect bone metabolism, or with missing data, were excluded from the analysis. RESULTS: The median intake of milk in the 1479 participants (mean age, 73.0 ± 5.1 years) was one glass of milk per day. Bone turnover markers showed a decreasing trend (p < 0.05) and aBMD at TH (p = 0.0019) and FN (p = 0.0057) and TBS (p = 0.0017) showed increasing trends with greater milk intake after adjusting for demographic and behavioral confounding factors. This association was attenuated after further adjusting for nutrient intake, in particular, calcium intake. CONCLUSIONS: Greater milk intake was associated with lower bone turnover, higher aBMD, and higher TBS in community-dwelling elderly Japanese men.

Upregulation of relaxin receptors in the PDL by biophysical force.

OBJECTIVES: We have previously reported that relaxin (Rln) expression from the ovary is upregulated by orthodontic tooth movement. This study was performed to test the hypothesis that Rln family peptides (Rxfps), the G-protein-coupled Rln receptor, is induced in periodontal ligament (PDL) cells to modulate the molecules involved in periodontal tissue remodeling while applying biophysical force. MATERIALS AND METHODS: Rats were implanted with orthodontic appliances to investigate changes to Rxfps in vivo. An in vitro biophysical force analysis was performed to measure the level of Rxfp 1 messenger RNA (mRNA) in primary human PDL cells. RESULTS: The levels of Rxfp 2 transcription and translation increased in a time-dependent manner during tooth movement. Rxfp 2 was localized in the PDL by immunofluorescence. In vitro analyses revealed that the level of Rxfp 1 mRNA in PDL cells increased significantly with both compression and tension force. The levels of matrix metalloproteinase (MMP)-1, MMP-2, interleukin-6, and vascular endothelial growth factor mRNA, which are important for periodontal tissue remodeling, also changed under force application and Rln treatment. CONCLUSIONS: PDL cells responded to Rln to modulate effector molecules for periodontal tissue remodeling by upregulating Rxfps expression under a biophysical force. CLINICAL RELEVANCE: Rln and Rxfps may serve as a PDL turnover molecule complex to control orthodontic tooth movement.

First Report of Peanut mottle virus Infecting Soybean in South Korea.

In July 2013, soybean (Glycine max) plants at the research field in Daegu, South Korea, showed virus-like symptoms, such as mosaic, mottle, yellowing, and stunting. Overall, there were approximately 1% of soybean plants that showed these symptoms. Sixteen soybean samples were collected based on visual symptoms and subjected to laboratory characterization. Total RNA was extracted from each sample with the Tri Reagent (Molecular Research Center, Cincinnati, OH) and cDNA was synthesized using random N25 primer with RevertAid Reverse Transcriptase (Thermo Scientific, Waltham, MA), according to the manufacturers' instructions. All samples were tested by PCR with Prime Taq Premix (2X) (GeNet Bio, Daejeon, Korea) and primer sets specific to Soybean mosaic virus (SMV; 5'-CATATCAGTTTGTTGGGCA-3' and 5'-TGCCTATACCCTCAACAT-3'), Peanut stunt virus (PSV; 5'-TGACCGCGTGCCAGTAGGAT-3' and 5'-AGGTDGCTTTCTWTTGRATTTA-3'), Soybean yellow mottle mosaic virus (SYMMV; 5'-CAACCCTCAGCCACATTCAACTAT-3' and 5'-TCTAACCACCCCACCCGAAGGATT-3'), and Soybean yellow common mosaic virus (SYCMV; 5'-TTGGCTGAGAGGAGTGGCTT-3' and 5'-TGCGGTCGTGTAGTCAGTG-3'). Among 16 samples tested, five were positive for SMV and two for SYMMV. Three samples were found infected by both SMV and SYMMV and four by both SMV and PSV. Since two of the symptomatic samples were not infected by viruses described above, a pair of primers specific to Peanut mottle virus (PeMoV; 5'-GCTGTGAATTGTTGTTGAGAA-3' and 5'-ACAATGATGAAGTTCGTTAC-3') was tested (1). All 16 samples were subjected to RT-PCR with primers specific to PeMoV. Four were found positive for PeMoV. Two of them were already found infected with SYMMV. In order to identify the complete nucleotide sequences of PeMoV coat protein (CP), another primer set (5'-TGAGCAGGAAAGAATTGTTTC-3' and 5'-GGAAGCGATATACACACCAAC-3') was used. RT-PCR product was cloned into RBC TA Cloning Vector (RBC Bioscience, Taipei, Taiwan) and the nucleotide sequence of the insert was determined by Macrogen (Seoul, Korea). CP gene of the PeMoV (GenBank Accession No. KJ664838) showed the highest nucleotide sequence identity with PeMoV isolate Habin (KF977830; 99% identity), and the highest amino acid identity with GenBank Accession No. ABI97347 (100% identity). In order to fulfill Koch's postulates, several G. max cv. Williams 82 were inoculated with the extracts of PeMoV-infected leaf tissue. At 14 days post-inoculation, plants showed systemic mottle symptoms. These symptomatic plants were subjected to RT-PCR, and the nucleotide sequences of the PCR product were found identical to that of the virus in the inoculum. To our knowledge, this is the first report of soybean-infecting PeMoV, a member of the genus Potyvirus in the family Potyviridae, in South Korea. Reference: (1) R. G. Dietzgen et al. Plant Dis. 85:989, 2001.

Lipopolysaccharide-induced indoleamine 2,3-dioxygenase expression in the periodontal ligament.

BACKGROUND AND OBJECTIVE: Indoleamine 2,3-dioxygenase (IDO) is a tryptophan-oxidizing enzyme with immune-inhibitory effects. The aim of this study was to investigate the expression of IDO by lipopolysaccharide (LPS), a component of gram-negative bacteria, in human periodontal ligament (PDL) cells. MATERIAL AND METHODS: Human PDL cells and gingival fibroblasts (GFs) were prepared from explants of human PDLs and from gingival tissues of clinically healthy donors, respectively. Real-time RT-PCR, western blotting and the IDO enzyme assay were performed to determine the expression of IDO following LPS treatment of cells. LPS was injected into mice tail veins to evaluate the effects of LPS in vivo in the maxillary first molar. Immunofluorescence staining and histological analysis were followed to localize IDO in mouse PDL. RESULTS: The level of expression of IDO mRNA in primary human PDL cells after LPS treatment was increased in a dose-dependent manner, reaching a peak 8 h after LPS treatment. The expression and activities of IDO protein were significantly increased in comparison with those of the control. In addition, the increased production of kynurenine in culture medium was observed 72 h after LPS treatment. In the immunofluorescence findings, stronger immunoreactivities were shown in PDL than in gingival tissues in the maxillae. In accordance with the immunofluorescence findings, LPS treatment induced a strong up-regulation of IDO mRNA in human PDL cells, whereas human GFs showed only a weak response to LPS. CONCLUSION: These results clearly show that IDO was induced by LPS in primary human PDL cells, suggesting that PDL might be involved in the regulation of oral inflammatory disease.

Association between vitamin K intake from fermented soybeans, natto, and bone mineral density in elderly Japanese men: the Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) study.

SUMMARY: A cross-sectional analysis of 1,662 community dwelling elderly Japanese men suggested that habitual natto intake was significantly associated with higher bone mineral density (BMD). When adjustment was made for undercarboxylated osteocalcin levels, this association was insignificant, showing the natto-bone association to be primarily mediated by vitamin K. INTRODUCTION: Low vitamin K intake is associated with an increased risk of hip fracture, but reports have been inconsistent on its effect on BMD. Our first aim was to examine the association between BMD and intake of fermented soybeans, natto, which contain vitamin K1 (20 µg/pack) and K2 (380 µg/pack). Our second aim was to examine the association between undercarboxylated osteocalcin (ucOC), a biomarker of vitamin K intake, and BMD to evaluate the role of vitamin K in this association. METHODS: Of the Japanese men aged ≥65 years who participated in the baseline survey of the Fujiwara-kyo Osteoporosis Risk in Men study, 1,662 men without diseases or medications known to affect bone metabolism were examined for associations between self-reported natto intake or serum ucOC levels with lumbar spine or hip BMD. RESULTS: The subjects with greater intake of natto showed significantly lower level of serum ucOC. Analysis after adjustment for confounding variables showed an association of greater intake of natto with both significantly higher BMD and lower risk of low BMD (T-score < -1 SD) at the total hip and femoral neck. This association became insignificant after further adjustment for ucOC level. CONCLUSION: Habitual intake of natto was associated with a beneficial effect on bone health in elderly men, and this association is primarily due to vitamin K content of natto, although the lack of information on dietary nutrient intake, including vitamin K1 and K2, prevented us from further examining the association.

Serum undercarboxylated osteocalcin levels are inversely associated with glycemic status and insulin resistance in an elderly Japanese male population: Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) Study.

SUMMARY: Recent animal studies have demonstrated that undercarboxylated osteocalcin upregulates insulin secretion via osteoblast-insulin signaling. However, it remains unclear whether such a pathway exists in humans. This study showed that serum undercarboxylated osteocalcin levels were inversely associated with fasting plasma glucose, hemoglobin A(1c), and homeostasis model assessment of insulin resistance (HOMA-IR) levels in community-dwelling elderly Japanese men. INTRODUCTION: Undercarboxylated osteocalcin (ucOC) was reported to increase insulin secretion and improve glucose tolerance via osteoblast-insulin signaling in animal-based studies. Whether this pathway also exists in humans is unknown. We aimed to clarify whether serum ucOC levels are associated with glycemic status and insulin resistance in the general Japanese population. METHODS: We included 2,174 Japanese men (≥65 years) who were able to walk without aid from others and lived at home in four cities of Nara Prefecture. We excluded participants with a history of diseases or medications that affect bone metabolism, other than type 2 diabetes mellitus (T2DM). Fasting plasma glucose, glycated hemoglobin A(1c), and HOMA-IR levels were determined as outcome measures. RESULTS: Of the 1,597 participants included in the analysis, both intact OC (iOC) and ucOC levels showed significant inverse correlations with all outcome measures, even after adjusting for potential confounders. Mean values of outcome measures showed a significant decreasing trend with higher quintiles of iOC or ucOC after adjusting for confounders. This trend remained significant for ucOC quintiles after further adjustment for iOC levels, but was not significant for iOC quintiles after adjusting for ucOC levels. These results were attenuated, but still apparent, after excluding participants receiving drug therapy for T2DM. CONCLUSIONS: Levels of ucOC, but not iOC, were inversely associated with glycemic index and insulin resistance in a population of Japanese men. These findings will need to be confirmed with longitudinal studies.

Optimization of electrotransformation conditions for Leuconostoc mesenteroides subsp. mesenteroides ATCC8293.

AIMS: To establish an efficient genetic transformation protocol for Leuconostoc species, methods for competent-cell preparation and electroporation conditions were optimized. METHODS AND RESULTS: Leuconostoc mesenteroides subsp. mesenteroides ATCC8293 cells were sequentially treated with penicillin G and lysozyme, and the plasmid pLeuCM was subsequently transformed into the cells. Our results demonstrated that transformation efficiencies were significantly increased (100-fold), and increased electric field strength also contributed to enhance transformation efficiency. Maximum transformation efficiency (1×10(4) or more transformants per µg DNA) was achieved when cells were grown in De Man, Rogosa, Sharpe (MRS) media containing 0·25 mol l(-1) sucrose and 0·8 µg ml(-1) penicillin G, followed by treatment with 600 U ml(-1) lysozyme and electroporation at a field strength of 10 kV cm(-1). When this protocol was used to transform pLeuCM into Leuc. mesenteroides, Leuconostoc gelidum, Leuconostoc fallax and Leuconostoc argentinun, successful transformations were obtained in all cases. Furthermore, this procedure was applicable to species belonging to other genera, including Lactobacillus plantarum, Pediococcus pentosaceus and Weissella confusa. CONCLUSIONS: The results demonstrate that the transformation efficiency for Leuconostoc spp. could be increased via optimization of the entire electroporation procedures. SIGNIFICANCE AND IMPACT OF THE STUDY: These optimized conditions can be used for the extensive genetic study and the metabolic engineering of not only Leuconostoc spp. but also different species of lactic acid bacteria.

Evaluation of selective enrichment broths and chromogenic media for Salmonella detection in highly contaminated chicken carcasses.

We evaluated the effectiveness of 2 selective enrichment broths, Rappaport-Vassiliadis Soy (RVS) and Muller-Kauffmann tetrathionate with novobiocin (MKTTn), for the isolation of Salmonella from chicken carcasses obtained from 3 different types of retail markets. We also compared a chromogenic agar, chromID Salmonella agar (SM-ID 2), with a classic plating medium, xylose lysine deoxycholate agar (XLD). Salmonella were isolated from 118 of the 180 samples (65.5%). Salmonella were detected in 105 samples (88%) plated on XLD and 111 samples (94%) plated on SM-ID 2 when RVS broth was used for enrichment, and 43 samples (36.4%) plated on XLD and 67 samples (56.8%) plated on SM-ID 2 when the MKTTn broth was used. The highest sensitivity was found in the RVS-XLD combination (0.99), followed by RVS-SM-ID 2 (0.97). The specificity of the RVS-SM-ID 2 combination was the highest (0.89), but that of the MKTTn-XLD combination was zero. The results of this study indicate that the selective enrichment broths had a great effect on the sensitivity and specificity of plating media, and our study confirms that the RVS broth is the most suitable enrichment for the investigation of Salmonella in chicken carcasses. This observation suggests that use of RVS broth for selective enrichment and SM-ID 2 for selective isolation may be the best combination to determine the presence of Salmonella in chicken carcasses.

Differential expression of cxcl-14 during eruptive movement of rat molar germs.

Tooth eruption at the early postnatal period is strictly controlled by the molecules secreted mainly from follicular tissues, which recruit monocytes for osteoclast formation. In this study, it was hypothesized that different molecules can be expressed according to the stages of tooth eruption. Rat molar germs together with follicles were extracted and DD-PCR was performed from the root formation stage 2nd molars germs (after eruptive movement) and cap stage 3rd molar germs (before movement) at postnatal day 9. Cxcl-14, a potent chemoattractant, was detected as one of the differentially expressed molecules from DD-PCR. Its expression increased significantly at the root formation stage, compared with the cap or crown formation stage at both transcription and translation levels. The expression patterns of cxcl-14 were consistent with those of MCP-1 and CSF-1, and opposite to OPG. Immunofluorescence showed that cxcl-14 was localized in the dental follicular tissues only at the root formation stage overlaying the proximo-occlusal region of the molar germs. Many osteoclasts appeared on the surface of the alveolar bone which overlayed the occlusal region of the root formation stage 2nd molar germs and underwent resorption. Cxcl-14 expression was reduced considerably at both the translation and transcription levels by an alendronate treatment. These results suggest that cxcl-14 may be implicated in the formation of the eruptive pathway of tooth germs via osteoclastogenesis.

Impact of smoking on bone mineral density and bone metabolism in elderly men: the Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) study.

UNLABELLED: Our cross-sectional analysis of 1,576 men aged ≥65 years examined smoking effects on bone status. Number of smoking years was associated with decreased bone mineral density (BMD), after adjusting for age, height, weight, and number of cigarettes smoked daily. Smoking did not affect biochemical marker serum values for bone turnover. INTRODUCTION: The impact of smoking on bone status in men has not been conclusively established. We examined how smoking and its cessation influence bone status and metabolism in men. METHODS: We analyzed 1,576 men among a baseline survey of Japanese men aged ≥65 years, the Fujiwara-kyo Osteoporosis Risk in Men study, conducted during 2007-2008. RESULTS: Lumbar spine (LS) BMD values among never, former, and current smokers were 1.045 ± 0.194, 1.030 ± 0.189, and 1.001 ± 0.182 g/cm(2) (P = 0.005), respectively, while total hip (TH) BMD values were 0.888 ± 0.120, 0.885 ± 0.127, and 0.870 ± 0.124 (P = 0.078), respectively. The significant trend for LS BMD remained after adjusting for the covariates; age, height, weight, physical activity, milk consumption, and drinking habit (P = 0.036). Among never and ever (current and former) smokers, LS and TH BMD decreased with the number of pack years or the number of smoking years, respectively, adjusted for those covariates. Among ever smokers, LS and TH BMD decreased with the number of smoking years after adjusting for age, height, weight, and number of cigarettes smoked daily. Smoking did not reveal significant effect for serum osteocalcin or tartrate resistant acid phosphatase isoenzyme 5b. CONCLUSION: The impact of smoking on bone status is mainly associated with the number of smoking years in elderly men.

Antimicrobial susceptibility of coagulase-negative staphylococci isolated from bovine mastitis between 2003 and 2008 in Korea.

A total of 1,444 coagulase-negative staphylococci (CNS) isolates from bovine mastitic milk samples collected during 2003-2008 in Korea were identified to the species level. Of 14 species identified, S. simulans, S. haemolyticus, and S. sciuri accounted for over 60% of the isolates. All the CNS isolates were tested for susceptibility to eight antimicrobials commonly used in dairy cattle. With a few exceptions, similar resistance patterns were observed among the CNS species: penicillin and ampicillin showed the lowest activity while amikacin, cephalothin, and gentamycin were highly effective. About 39% (557/1,444) of the CNS isolates were pan-susceptible, while 12% (175/1,444) showed resistance to four or more antimicrobials tested.

Prevalence and antimicrobial susceptibility of gram-negative bacteria isolated from bovine mastitis between 2003 and 2008 in Korea.

The objective of this study was to assess trends in the prevalence and distribution of gram-negative bacteria isolated from bovine mastitis and their antimicrobial susceptibilities during a 6-yr period between 2003 and 2008 in Korea. Escherichia coli, Pseudomonas fluorescens, Klebsiella pneumoniae, Enterobacter cloacae, Acinetobacter lwoffi/junii, Pseudomonas aeruginosa, and Serratia marcescens were the most commonly observed pathogens during this period. Generally, gram-negative bacteria showed low susceptibilities to most of the antimicrobials tested in this study, except amikacin and gentamicin. Although these 2 aminoglycosides were broadly active against gram-negative bacteria, less than half of those bacteria showed susceptibilities to streptomycin. The beta-lactams, except piperacillin, had the lowest activity among antimicrobials tested in this study. Susceptibilities to chloramphenicol and trimethoprim were fairy high in all genera of gram-negative bacteria, except Acinetobacter spp. and Pseudomonas spp., whereas relatively high resistance to tetracycline was observed uniformly among gram-negative bacteria. There was no significant change in the prevalence of bacterial and the proportion of antimicrobial resistance among gram-negative bacteria isolates during a 6-yr period.

First Report of Soybean yellow mottle mosaic virus in Soybean in North America.

Soybean yellow mottle mosaic virus (SYMMV) is a soybean-infecting virus recently discovered in Korea that initially induces bright yellow mosaic on leaves followed by stunting and reduced growth of older leaves (1). Nucleotide sequence analysis of genomic RNA of the Korean SYMMV isolate suggested that the virus is a new member of the genus Carmovirus in the family Tombusviridae. To determine whether SYMMV is present in the United States, single leaflets were collected without regard for symptoms from 7 to 10 plants in each of 136 plots in August 2008 from a research field in Stoneville, MS that contained 16 plant introductions (including five from Korea) and 'Williams 82'. Samples were grouped into 10 pools of 100 leaves from which total RNA was extracted with the Qiagen RNeasy Plant Mini Kit (Germantown, MD), reverse transcribed, and amplified with SuperScript III Platinum SYBR Green One-Step Quantitative Real-time Reverse Transcriptase-PCR Kit (Invitrogen, Carlsbad, CA) and two pairs of oligonucleotide primers (5'-CGTCTGCCAGGGTTTAATACTA-3', and 5'-GATTAGCATGTCAGGGTGGTCG-3'; and 5'-ACTGAGTCCCCTGCTTAT-3' and 5'-CATCACTAGCGTCYGGATCA-3') that were designed from regions conserved between SYMMV and Cowpea mottle virus (CPMoV; a related and seed-transmitted carmovirus). Six 100-leaflet pools were positive with both primer sets and four pools were negative with both primer sets. Total RNA extracted from one positive pool was reverse transcribed using SuperScript II reverse transcriptase and a primer complementary to nt 4,000 to 4,009 of the SYMMV genome and amplified using iProof DNA polymerase (Bio-Rad, Hercules, CA) as two overlapping DNA fragments using primers corresponding to nt 1 to 21 and complementary to nt 3,483 to 3,508 and corresponding to nt 3,366 to 3,391 and complementary to nt 4,000 to 4,009. DNA fragments were sequenced using a BigDye Terminator Cycle Sequencing Kit and ABI 3730XL capillary sequencers (Applied Biosystems, Foster City, CA). The 4,009-nt sequence of the Mississippi SYMMV isolate (GenBank Accession No. FJ707484) was 96% identical to the Korean SYMMV isolate and 65% identical to CPMoV. Because of the sampling techniques used, it was not possible to associate SYMMV-positive plants with disease symptoms in Mississippi. To our knowledge, this is the first report of SYMMV in North America. Reference: (1) M. Nam et al. Online publication. doi:10.1077/s00705-009-0480. Arch. Virol., 2009.

Novel black soy peptides with antiobesity effects: activation of leptin-like signaling and AMP-activated protein kinase.

OBJECTIVE: To investigate the mechanisms underlying the antiobesity effects of a novel isoflavone-free peptide mixture (BSP) derived from black soybean. DESIGN: Long-term effects of BSP were evaluated in diet-induced obese (DIO) mice fed a high-fat (HF) diet without or with BSP (2, 5 or 10% of energy) for 13 weeks, or for 8 weeks in combination with exercise. Acute effects of BSP on food intake and body weight in rats and leptin-deficient ob/ob mice were evaluated. Cell culture models or tissue extracts were used to investigate the mechanisms underlying the antiobesity effect. MEASUREMENT: Total food intake, body weight gain, white adipose tissue (WAT) mass, plasma concentrations of leptin, adiponectin, cholesterol and triglyceride were measured. Janus kinase 2 (JAK2)-dependent signal transducers and activators of the transcription 3 (STAT3) phosphorylation and AMP-activated protein kinase (AMPK) activity were determined using Western-blot in cultured cells or tissue extracts. RESULTS: DIO mice fed an HF diet with BSP (2, 5 or 10%) for 13 weeks gained less body weight (21.4, 19.8 or 17.1 g, respectively) than the mice fed an HF diet without BSP (22.6 g) concurrent with inhibition of total food intake in a dose-dependent manner. BSP also significantly decreased food intake in rats and leptin-deficient ob/ob mice. The highest dose of BSP (10%) significantly elevated the plasma adiponectin and decreased plasma triglyceride. BSP activated JAK2-dependent STAT3 in a cell model, and elevated the level of hypothalamic phospho-STAT3 in ob/ob mice. BSP also phosphorylated AMPK and acetyl-CoA carboxylase of C2C12 myocytes in a dose-dependent manner. The antiobesity effect was augmented by low-intensity wheel-based exercise. In exercised mice, BSP significantly decreased periepididymal WAT mass and body weight gain. CONCLUSION: These results provided evidences that BSP decreased appetite and HF diet-induced body weight gain particularly in combination with exercise, through leptin-like STAT3 phosphorylation and AMPK activation.

Can we predict spontaneous capsule passage after retention? A nationwide study to evaluate the incidence and clinical outcomes of capsule retention.

BACKGROUND AND STUDY AIMS: Although capsule endoscopy has become a central diagnostic tool for small-bowel evaluation, retention of a capsule remains a major concern. This study attempted to investigate the incidence and clinical outcomes of capsule retention, and to determine the factors predictive of spontaneous capsule passage after retention. PATIENTS AND METHODS: Through a nationwide multicenter survey, we retrospectively reviewed the records of 1291 patients who had a capsule endoscopy between February 2002 and July 2006 in Korea. Clinical and procedural characteristics and postprocedural outcomes were analyzed for the cases with capsule retention. RESULTS: Capsule retention occurred in 2.5 % of total cases (32/1291). The major diseases accompanying capsule retention were Crohn's disease, malignant tumors, and tuberculous enterocolitis, in decreasing order. In 11 of the 32 patients (34.4 %), early surgical or endoscopic interventions were instituted for diagnosis or treatment of diseases before retention symptoms developed. The remaining 21 (65.6 %) patients initially received medical treatments. Of these, 10 (31.3 %) ultimately underwent surgical intervention due to the development of symptoms of intestinal obstruction or medical treatment failure. The other 11 (34.4 %) eventually passed the capsule. The presence of a larger lumen diameter (greater than two-thirds of the capsule diameter) at the stricture site was associated with spontaneous passage. CONCLUSIONS: Our large-scale study suggests that retention occurs infrequently during capsule endoscopy. Moreover, a retained capsule might indicate the best intervention for the offending pathology, or it may spontaneously pass in the long run, particularly in patients with less small bowel stricture.

Application of a new portable microscopic somatic cell counter with disposable plastic chip for milk analysis.

The somatic cell count (SCC) is one of the international standards for monitoring milk quality, and it is a useful indicator of mastitis. The current reference method for determining the SCC in raw milk is direct microscopic analysis, but this method requires well-trained staff to maintain its accuracy and reproducibility. To overcome these inconveniences, we developed a portable system (the C-reader system) that utilizes the capillary flow of a microfluidic chamber by surface modification of the hydrophilicity. The microfluidic technology of disposable microchips allows for low consumption of reagents, and a combination of ready-to-use reagents makes the daily work easier. The repeatability test of the C-reader using 10 composite bovine milk samples satisfied the recommended values for SCC equipment. In addition, an acceptable accuracy level of the natural logarithmic-transformed SCC [ln(SCC/1,000): +/- 0.059 to 0.112] was achieved using composite raw milk samples and various somatic cell standard solutions from the American Eastern Laboratory and the Korean National Veterinary Research and Quarantine Service. After testing 875 composite milk samples, the C-reader showed a high correlation coefficient (R2 = 0.935 to 0.964) and a low mean difference value in log-transformed SCC (-0.088 to 0.004) compared with 3 automatic commercialized somatic cell counters (Fossomatic 4000, Somacount 150, and Somascope). In conclusion, the C-reader system is a new, easy-to-use automatic on-farm method with acceptable repeatability and accuracy for measuring SCC in large dairies and smaller laboratories.