追加1. 胃切除術後の鼻腔ゾンデ早期抜去について(シンポジウム 中山式切除術をめぐって,第473回千葉医学会例会,第4回佐藤外科例会)

Additional file 4. Proteomic data of strains wild-type, AG553, AG553, AG601 and LL1210 of Clostridium thermocellum

Identification of sarV (SA2062), a New Transcriptional Regulator, Is Repressed by SarA and MgrA (SA0641) and Involved in the Regulation of Autolysis in Staphylococcus aureus

The expression of genes involved in the pathogenesis of Staphylococcus aureus is known to be controlled by global regulatory loci, including agr, sarA, sae, arlRS, lytSR, and sarA-like genes. Here we described a novel transcriptional regulator called sarV of the SarA protein family. The transcription of sarV is low or undetectable under in vitro conditions but is significantly augmented in sarA and mgrA (norR or rat) (SA0641) mutants. The sarA and mgrA genes act as repressors of sarV expression, as confirmed by transcriptional fusion and Northern analysis data. Purified SarA and MgrA proteins bound specifically to separate regions of the sarV promoter as determined by gel shift and DNase I footprinting assays. The expression of 19 potential target genes involved in autolysis and virulence, phenotypes affected by sarA and mgrA, was evaluated in an isogenic sarV mutant pair. Our data indicated that the sarV gene product played a role regulating some virulence genes and more genes involved in autolysis. The sarV mutant was more resistant to Triton X-100 and penicillin-induced lysis compared to the wild type and the sarA mutant, whereas hyperexpression of sarV in the parental strain or the sarV mutant rendered the resultant strain highly susceptible to lysis. Zymographic analysis of murein hydrolase activity revealed that inactivation of the sarV gene results in decreased extracellular murein hydrolase activity compared to that of wild-type S. aureus. We propose that sarV may be part of the common pathway by which mgrA and sarA gene products control autolysis in S. aureus

Identifying promoters for gene expression in Clostridium thermocellum

A key tool for metabolic engineering is the ability to express heterologous genes. One obstacle to gene expression in non-model organisms, and especially in relatively uncharacterized bacteria, is the lack of well-characterized promoters. Here we test 17 promoter regions for their ability to drive expression of the reporter genes β-galactosidase (lacZ) and NADPH-alcohol dehydrogenase (adhB) in Clostridium thermocellum, an important bacterium for the production of cellulosic biofuels. Only three promoters have been commonly used for gene expression in C. thermocellum, gapDH, cbp and eno. Of the new promoters tested, 2638, 2926, 966 and 815 showed reliable expression. The 2638 promoter showed relatively higher activity when driving adhB (compared to lacZ), and the 815 promoter showed relatively higher activity when driving lacZ (compared to adhB). Keywords: Promoter, Copy number, Structural instability, Rolling circle replication, lacZ, adh

Deletion of the hfsB gene increases ethanol production in Thermoanaerobacterium saccharolyticum and several other thermophilic anaerobic bacteria

Hettich, Robert L/0000-0001-7708-786X; Lynd, Lee R/0000-0002-5642-668X; BELDUZ, Ali Osman/0000-0003-2240-7568; Olson, Daniel/0000-0001-5393-6302; Giannone, Richard/0000-0001-8551-0138; Eminoglu, Aysenur/0000-0003-1693-6332; Murphy, Sean/0000-0001-9268-6684WOS: 000416866000002PubMed: 29213322Background: With the discovery of interspecies hydrogen transfer in the late 1960s (Bryant et al. in Arch Microbiol 59:20-31, 1967), it was shown that reducing the partial pressure of hydrogen could cause mixed acid fermenting organisms to produce acetate at the expense of ethanol. Hydrogen and ethanol are both more reduced than glucose. Thus there is a tradeoff between production of these compounds imposed by electron balancing requirements; however, the mechanism is not fully known. Results: Deletion of the hfsA or B subunits resulted in a roughly 1.8-fold increase in ethanol yield. the increase in ethanol production appears to be associated with an increase in alcohol dehydrogenase activity, which appears to be due, at least in part, to increased expression of the adhE gene, and may suggest a regulatory linkage between hfsB and adhE. We studied this system most intensively in the organism Thermoanaerobacterium saccharolyticum; however, deletion of hfsB also increases ethanol production in other thermophilic bacteria suggesting that this could be used as a general technique for engineering thermophilic bacteria for improved ethanol production in organisms with hfs-type hydrogenases. Conclusion: Since its discovery by Shaw et al. (JAMA 191:6457-64, 2009), the hfs hydrogenase has been suspected to act as a regulator due to the presence of a PAS domain. We provide additional support for the presence of a regulatory phenomenon. in addition, we find a practical application for this scientific insight, namely increasing ethanol yield in strains that are of interest for ethanol production from cellulose or hemicellulose. in two of these organisms (T. xylanolyticum and T. thermosaccharolyticum), the ethanol yields are the highest reported to date.Scientific and Technological Research Council of Turkey (TUBITAK-BIDEB)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [2214/A]; Office of Biological and Environmental Research in the DOE Office of Science; Office of Science of the U.S. Department of EnergyUnited States Department of Energy (DOE) [DE-AC02- 05CH11231]; U.S. Department of EnergyUnited States Department of Energy (DOE) [DE-AC05-00OR22725]In this work, AE was supported by the Scientific and Technological Research Council of Turkey (TUBITAK-BIDEB 2214/A International Doctoral Research Fellowship Program) with a scholarship. the BioEnergy Science Center is a U.S. Department of Energy Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science. the genomic resequencing work conducted by the U.S. Department of Energy Joint Genome Institute, a DOE Office of Science User Facility, is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02- 05CH11231. This manuscript has been authored by Dartmouth College under Contract No. DE-AC05-00OR22725 with U.S. Department of Energy. the US Government retains and the publisher, by accepting the article for publication, acknowledges that the US Government retains a non-exclusive, paid-up, irrevocable worldwide license to publish or reproduce the published form of this manuscript or allow others to do so, for US Government purposes

Ethanol tolerance in engineered strains of Clostridium thermocellum

Abstract Clostridium thermocellum is a natively cellulolytic bacterium that is promising candidate for cellulosic biofuel production, and can produce ethanol at high yields (75–80% of theoretical) but the ethanol titers produced thus far are too low for commercial application. In several strains of C. thermocellum engineered for increased ethanol yield, ethanol titer seems to be limited by ethanol tolerance. Previous work to improve ethanol tolerance has focused on the WT organism. In this work, we focused on understanding ethanol tolerance in several engineered strains of C. thermocellum. We observed a tradeoff between ethanol tolerance and production. Adaptation for increased ethanol tolerance decreases ethanol production. Second, we observed a consistent genetic response to ethanol stress involving mutations at the AdhE locus. These mutations typically reduced NADH-linked ADH activity. About half of the ethanol tolerance phenotype could be attributed to the elimination of NADH-linked activity based on a targeted deletion of adhE. Finally, we observed that rich growth medium increases ethanol tolerance, but this effect is eliminated in an adhE deletion strain. Together, these suggest that ethanol inhibits growth and metabolism via a redox-imbalance mechanism. The improved understanding of mechanisms of ethanol tolerance described here lays a foundation for developing strains of C. thermocellum with improved ethanol production

MOESM8 of Deletion of the hfsB gene increases ethanol production in Thermoanaerobacterium saccharolyticum and several other thermophilic anaerobic bacteria

Additional file 8: Figure S1. Confirmation of hydrogenase deletions in HKO strains

L'Auto-vélo : automobilisme, cyclisme, athlétisme, yachting, aérostation, escrime, hippisme / dir. Henri Desgranges

03 juin 19161916/06/03 (A17,N5612)

Late Remodeling Around a Proximally HA-coated Tapered Titanium Femoral Component

Most bone remodeling is thought to occur within the first few years after THA. Loss of bone density later may be associated with stress shielding or normal bone loss of aging. We evaluated remodeling changes over time with a proximally hydroxyapatite-coated tapered titanium stem. We evaluated plain radiographs of 143 hips for cancellous condensation, cortical hypertrophy, cortical porosis, cortical index, and canal fill at early postoperative, 5, 10, and 15 years. Average age was 51 years at THA; 69 patients (77 hips) (53%) were women; and 102 hips (71%) had primary osteoarthrosis. Based on radiographic findings at 15 years, hips were divided into three subgroups: 43 (30%) demonstrated minimal remodeling changes; 53 (37%) demonstrated cortical hypertrophy evident before 5 years; and 47 (33%) demonstrated additional late remodeling and cortical porosis, most often after 10 years. Hips with poorer bone (Dorr Types B or C) and, when including only hips with osteoarthrosis, more female hips had cortical porosis at 15 years. Late radiographic changes in patients with porosis appear more similar to that associated with an extensively rather than proximally coated stem. Whether continued bone adaptation and bone loss of aging will eventually threaten implant stability is unknown, but at 15 years, all 143 implants remained well fixed and clinically asymptomatic