Expression of human thromboxane synthase using a baculovirus system

AbstractHuman thromboxane (TX) synthase (EC 5.3.99.5) was produced by the baculovirus expression system using cDNA encoding human TX synthase [(1991) Biochem. Biophys. Res. Commun. 78, 1479-1484]. A recombinant baculovirus TXS7 was expressed in Spodoptera frugiperda Sf9 insect cells. The expressed protein was recognized by monoclonal antibody, Kon 7 raised against human TX synthase [(1990) Blood 76, 80-85]. The recombinant TX synthase catalyzed the conversion of prostaglandin (PG) H2 to TXA2 and 12-hydroxy-heptadecatrienoic acid (HHT). Both conversions of PGH2 to TXA2 and HHT by the expressed TX synthase were completely inhibited by a specific TX synthase inhibitor, OKY-046 (5 μM)

Expression of human thromboxane synthase using a baculovirus system

AbstractHuman thromboxane (TX) synthase (EC 5.3.99.5) was produced by the baculovirus expression system using cDNA encoding human TX synthase [(1991) Biochem. Biophys. Res. Commun. 78, 1479-1484]. A recombinant baculovirus TXS7 was expressed in Spodoptera frugiperda Sf9 insect cells. The expressed protein was recognized by monoclonal antibody, Kon 7 raised against human TX synthase [(1990) Blood 76, 80-85]. The recombinant TX synthase catalyzed the conversion of prostaglandin (PG) H2 to TXA2 and 12-hydroxy-heptadecatrienoic acid (HHT). Both conversions of PGH2 to TXA2 and HHT by the expressed TX synthase were completely inhibited by a specific TX synthase inhibitor, OKY-046 (5 μM)

Perioperative Elevation in Cell-Free DNA Levels in Patients Undergoing Cardiac Surgery: Possible Contribution of Neutrophil Extracellular Traps to Perioperative Renal Dysfunction

Background. This study aimed to determine the perioperative change in serum double-strand DNA (dsDNA) as a marker potentially reflecting neutrophil extracellular trap concentration in samples from patients undergoing cardiac surgery and to analyze a relationship between serum dsDNA concentrations and perioperative renal dysfunction. Methods. Serum dsDNA concentrations in samples that were collected during a previously conducted, prospective, multicenter, observational study were measured. Eighty patients undergoing elective cardiac surgery were studied. Serum samples were collected at baseline, immediately after surgery, and the day after surgery (POD-1). Results. Serum dsDNA concentration was significantly increased from baseline (median, 398 ng/mL [interquartile range, 372–475 ng/mL]) to immediately after surgery (median, 540 ng/mL [437–682 ng/mL], p<0.001), and they were reduced by POD-1 (median, 323 ng/mL [256–436 ng/mL]). The difference in serum creatinine concentration between baseline and POD-1 was correlated with dsDNA concentration on POD-1 (rs=0.61, p<0.001). Conclusions. In patients undergoing cardiac surgery, serum dsDNA concentration is elevated postoperatively. Prolonged elevation in dsDNA concentration is correlated with perioperative renal dysfunction. Further large-scale studies are needed to determine the relationship between serum concentration of circulating dsDNA and perioperative renal dysfunction

嚥下障害のない高齢者に対する安全なコップの検討：異なるコップが頭頸部伸展角度へ及ぼす影響

研究報告Original articles　我々は先行研究において，若年健常者を対象に一般的なコップで，誤嚥しにくい安全なコップの形状を判断するための指標として模型を作製した．本研究では，高齢者に対しても模型が安全なコップの形状を判断するのに有効であるかを検討するため，当院入院中の嚥下障害がない高齢者28 名を対象に，安全と判断したコップと危険と判断したコップの2 種類を用いて最大頭頸部伸展角度について検討した．またMMSE の得点により群分けし，認知レベルの違いによる飲み方の違いがあるか検討した．危険と判断したコップは安全と判断したコップと比較して最大頭頸部伸展角度が有意に大きく，さらに認知機能低下者は，形状に合わせた飲み方に注意を向けることが出来ず，頭頸部が過伸展した状態で嚥下してしまうため，危険性が増加する傾向にあった．したがって，頭頸部を過伸展することなく飲めるコップの選定が重要であることに加えて，先行研究で作製した模型の有用性が示唆された．In our previous study we created test models out of regular cups to evaluate safe shapes of cups that can prevent swallowing errors by testing them on young healthy individuals. In this study, in order to examine whether those model cups were safe for individuals as well, we used two types of cups, one considered safe, the other considered unsafe, with 28 older hospital patients without dysphagia, and examined the cervical spine extension. We also studied, by grouping data according to MMSE scores, whether there are differences in the manner of drinking based on different cognitive capacities. The maximum extension angle of the head and neck area was significantly larger when a cup considered unsafe was used compared to a safe one. Furthermore, individuals with impaired cognitive abilities tended to face increased danger, as they were not able to adjust their way of drinking to the shape of the cup, and as a result, tried to swallow with their head and neck areas overly extended. Therefore, it is essential to select a type of cup which can prevent the user overly extending their head and neck area. We also found that our previously created test model was effective with geriatric individuals as well

Infiltration of M1, but not M2, macrophages is impaired after unilateral ureter obstruction in Nrf2-deficient mice

Chronic inflammation can be a major driver of the failure of a variety of organs, including chronic kidney disease (CKD). The NLR family pyrin domain-containing 3 (NLRP3) inflammasome has been shown to play a pivotal role in inflammation in a mouse kidney disease model. Nuclear factor erythroid 2-related factor 2 (Nrf2), the master transcription factor for anti-oxidant responses, has also been implicated in inflammasome activation under physiological conditions. However, the mechanism underlying inflammasome activation in CKD remains elusive. Here, we show that the loss of Nrf2 suppresses fibrosis and inflammation in a unilateral ureter obstruction (UUO) model of CKD in mice. We consistently observed decreased expression of inflammation-related genes NLRP3 and IL-1β in Nrf2-deficient kidneys after UUO. Increased infiltration of M1, but not M2, macrophages appears to mediate the suppression of UUO-induced CKD symptoms. Furthermore, we found that activation of the NLRP3 inflammasome is attenuated in Nrf2-deficient bone marrow–derived macrophages. These results demonstrate that Nrf2-related inflammasome activation can promote CKD symptoms via infiltration of M1 macrophages. Thus, we have identified the Nrf2 pathway as a promising therapeutic target for CKD

Comparison of the virulence of exopolysaccharide-producing Prevotella intermedia to exopolysaccharide non-producing periodontopathic organisms

<p>Abstract</p> <p>Background</p> <p>Evidence in the literature suggests that exopolysaccharides (EPS) produced by bacterial cells are essential for the expression of virulence in these organisms. Secreted EPSs form the framework in which microbial biofilms are built.</p> <p>Methods</p> <p>This study evaluates the role of EPS in <it>Prevotella intermedia </it>for the expression of virulence. This evaluation was accomplished by comparing EPS-producing <it>P. intermedia </it>strains 17 and OD1-16 with non-producing <it>P. intermedia </it>ATCC 25611 and <it>Porphyromonas gingivalis </it>strains ATCC 33277, 381 and W83 for their ability to induce abscess formation in mice and evade phagocytosis.</p> <p>Results</p> <p>EPS-producing <it>P. intermedia </it>strains 17 and OD1-16 induced highly noticeable abscess lesions in mice at 10⁷colony-forming units (CFU). In comparison, <it>P. intermedia </it>ATCC 25611 and <it>P. gingivalis </it>ATCC 33277, 381 and W83, which all lacked the ability to produce viscous materials, required 100-fold more bacteria (10⁹CFU) in order to induce detectable abscess lesions in mice. Regarding antiphagocytic activity, <it>P. intermedia </it>strains 17 and OD1-16 were rarely internalized by human polymorphonuclear leukocytes, but other strains were readily engulfed and detected in the phagosomes of these phagocytes.</p> <p>Conclusions</p> <p>These results demonstrate that the production of EPS by <it>P. intermedia </it>strains 17 and OD1-16 could contribute to the pathogenicity of this organism by conferring their ability to evade the host's innate defence response.</p

Physiological and Ultrastructural Studies on the Origin of Activator Calcium in Body Wall Muscles of Spoon Worms

To examine the origin of activator Ca and its translocation during contraction in body wall muscles (BWM) of spoon worms, Urechis unicinctus , physiological and ultrastructural studies, including cytochemistry, were performed. The potassium (K-) contracture tension was significantly reduced by the removal of external Ca, and by the application of Mn, La and verapamil. On the other hand, caffeine induced a prolonged contraction. The removal of Ca and Mg from the external solution, and the rapid cooling caused an irregular or oscillatory contraction. These results suggested that, in BWM fibers, the activator Ca is supplied partially from both external solution and intracellular Ca-accumulating structures. Ultrastructural observations revealed that the muscle fibers contain a relatively large amount of sarcoplasmic reticulum (SR). The fractional volume of the SR relative to the fiber volume was 2~5% in all fibers of three muscle layers. To demonstrate the Ca localization, the muscle fibers were fixed by pyroantimonate (PA) methods at resting and contracting states. In the resting fibers, the PA precipitates were exclusively localized in the SR and the inner surface of plasma membrane. On the other hand, in the contracting fibers, they were diffusely distributed in the central regions of myoplasm, and had disappeared from the SR and plasma membrane. X-ray microanalysis revealed that the PA precipitates contain Ca. With the results of physiological experiments, these results indicate that the activator Ca originates not only from the external solution, but also from the intracellular Ca-accumulating structures, the SR and the inner surface of plasma membrane.Full-Length Pape

短期大学生入学初期の生活習慣獲得の実態

Students these days have been said to be short of basic life habits forming. For students who major in nursing, welfare or early childhood education, it is very important to form basic life habits because they need to support other people\u27s lives in their future careers. We have found out that comparatively many students have acquired the basic life habits in hygiene, but relatively smaller percentage of students have formed the basic life habits in protection against infecdon, and excretion. We need to consider how we can make our students acquire basic life habits