Endothelin as a local regulating factor in the bovine oviduct

Endothelin (EDN) is a possible regulating factor of oviductal motility, which is important for the transport of gametes and embryo. To clarify the factors that control the secretion of EDN in the bovine oviduct, the expression of EDNs, EDN-converting enzymes (ECEs) and EDN receptors (EDNRs) were investigated. All isoforms of EDN (EDN1-3), ECE (ECE1 and ECE2) and EDNR (EDNRA and EDNRB) were immunolocalised in the epithelial cells of the ampulla and the isthmus. EDNRs were also immunolocalised in smooth-muscle cells. The mRNA expression of EDN2 and ECE2 was higher in cultured ampullary oviductal epithelial cells than in isthmic cells. The expression of EDN1, EDN2 and ECE2 in the ampullary tissue was highest on the day of ovulation. Oestradiol-17β increased EDN2 and ECE1 expression, while progesterone increased only ECE1 expression in cultured ampullary epithelial cells. These results indicate that EDNs are produced by epithelial cells and their target site is smooth-muscle and epithelial cells, and suggest that ovarian steroids are regulators of endothelin synthesis in ampullary oviductal epithelial cells

Usefulness of the waterless method for surgical hand washing：A comparison with the conventional scrub method

To evaluate the usefulness of the waterless hand washing method for surgical anti-sepsis, we conducted a microbial sampling study, comparing it to a conventional surgical scrub method. A total of 18 operating-room nurses were undertaking the following three-hand washing protocols : scrubbing with blushes using 4w/v% chlorhexidine gluconate (CHG) and also rubbing with CHG, followed by application of a 0.2w/v% CHG with ethanol (HS) preparation. (conventional method); rubbing with CHG and application of HS (two-stage surgical scrub method; TSS); rubbing with anti-septic soap and application of HS (waterless method; WL). Microbial sampling was conducted after hand washing using the glove juice method. No statistically significant differences in bacterial numbers were found among these three methods. The number of bacterially positive subjects was significantly higher in the conventional method than the TSS method. These results indicate that there are adverse effects of blush-scrubbing, as the detected bacteria were related to normal skin flora. As such, the WL method for hand anti-sepsis appears to be equivalent to the conventional surgical scrubbing method in terms of microbial detection. The WL method should therefore be introduced as a standard hand anti-sepsis method at the time of surgery because it is cost-effective as well as time-efficient

生食用ソラマメ(ファーベ)およびソラマメ種子の諸成分について

生食用ソラマメのファーベと陵西一寸ソラマメの成熟過程における成分分析ならびに数種のソラマメ種子の分析を行い比較検討した。ファーベと陵西一寸の可溶性糖含量はともに小粒が高く成熟するにつれて低下したが,小粒段階での糖質は主にスクロースでありほかには少量のグルコースとフルクトースが含まれている。成熟に伴い陵西一寸ではスタキオース,ベルパスコースが急増したが,スクロース含量は低下し,デンプン含量は増加した。ファーベでは成熟過程でもベルバスコースは検出されず,スクロース含量の低下も緩やかで,デンプン含量は陵西一寸と比べて明らかに低かった。遊離アミノ酸含量はファーベと陵西一寸はともに成熟に伴い減少するがファーベでは陵西一寸ほどの急減は見られず徐々に減少した。ファーベ,陵西一寸両方とも,どの成育段階でも最多のアミノ酸はアルギニンであり,その他の主なアミノ酸はアスパラギン酸,アスパラギン,グルタミン酸,アラニン,GABAであった。成熟により急減したアミノ酸は陵西一寸ではアルギニン,アスパラギン,アラニン,GABAであったが,ファーベではアルギニンとGABA以外は徐々に低下した。ファーベと陵西一寸のGABA含量は小粒程高く,ファーベでは(固形分中)1000mg%にも達した。ソラマメ種子の可溶性糖は主にベルパスコースとスクロースを含むが,大部分の種子はベルバスコース含量の方が高い。ソラマメ種子の最多遊離アミノ酸は未熟ソラマメと同様アルギニンが主である。いずれの種子もGABA含量は激減している。ファーベは糖質組成分ヤアミノ酸の分析データなどから陵西一寸とは異なる性状のソラマメであり,生食に適していることを確認した。This study was performed to analyze the various components of seeds of immature fave and ryousai-issun (broad beans) at various stages of maturation, and analysis was also performed on several types of seeds for comparison purposes. The water-soluble sugar contents of fave and ryousai-issun were rather high, but decreased during maturation. The major components were sucrose. For ryousai-issun, with maturation, the contents of stachyose, verbascose increased rapidly, at the same time the content of sucrose decreased and the starch content increased. For fave, during the maturation process, verbascose was also not detected, the decrease of the sucrose content was rather gradual, and the starch content was less than that of the ryousai-issun. For both fave and ryousai-issun, free amino acids decreased with maturation, but unlike the rapid decrease for ryousai-issun, the decrease for fave was more gradual. For both fave and ryousai-issun, for all stages of maturation, the major content of amino acid was arginine while the other major amino acids included aspartic acid, asparagine, glutamic acid, alanine and GABA. For ryousai-issun, with maturation, the following amino acids decreased rapidly: arginine, asparagine, alanine and GABA. For fave, amino acids except arginine and GABA decreased gradually. For both fave and ryousai-issun, the content of GABA was higher with smaller beans, and for fave 1000mg% was detected (solid content). For ryousai-issun mature seeds, water-soluble sugar included verbascose and sucrose, but for most of the seeds, the content of verbascose was higher. For ryousai-issun seeds, just like the immature seeds, the main component of free amino acid was arginine. GABA content decreased rapidly. From the analytical data on the sugar and amino acids contents, fave proved to be different from ryousai-issun, and it was confirmed that raw fave was considered to be suitable for consumption

有用豆類の種実における可溶性糖質および遊離アミノ酸組成について

Twenty available samples belonging to 7 genera of peas and beans, some of which synthesize and accumulate starch, while others produce a large amount of fatty acids and proteins, were used for the analysis of soluble sugars and free amino acids, in order to distinguish the characteristics of the various peas and beans. The soluble sugars, including Glucose,Fructose,Sucrose,Raffmose,Stachyose,Verbascose were detected. Sucrose,Stachyose,Verbascose were verified to be the major components. The contents of the soluble sugars and the composition of the major-component sugars varied depending on the type of peas and beans. In Phaseolus vulgaris (Common bean) and Vigna angularis (Azuki bean), the composition of Stachyose was high, in Glycine max (Soy bean), Sucrose was high. In Pisum sativum (Pea) and Vicia fava (Broad bean), the composition of Verbascose was high. The compositions of the three types of sugars and the ratios to the overall contents seemed to be the characteristics of the peas and beans. Phaseolus sp. (Kidney bean, Lima bean), Vicia faba and Cicer arietinum etc. generally contained a high content of total free amino acids, with the highest of 1000 mg% for Kintoki mame. Compared to this, Vigna sp. (Azuki bean, Green gram, Rice bean, Cowpea) contained the lowest content, with about one third the value of the Green peas. Five types of amino acids found in high contents for each type of pea or bean include Asp, AspNfh, Glu, Pro, Arg. Glu was found in 7 samples examined. Vicia faba has high Arg, Phaseolus vulgaris and Phaseolus lunatus have high Glu, Glycine max has high Pro, showing that the content of amino acids differed in different peas and beans. For the total content of the \u27taste-related\u27 amino acids (Asp,Glu), Phaseolus vulgaris (Kintoki mame) showed the highest value, while Glycine max showed the lowest. As compared to the high contents of Glu and Cys of Phaseolus vulgaris, Glycine max showed a low content of Glu, but a high content of Pro. The amino acids composition of Phaseolus vulgaris and Glycine max differed largely from the other peas and beans. The total free amino acids contents and Pro of Glycine max and Cicer arietinum were high, and the sum of Pro and Arg of these two genera amounted to about 70% of the total free amino acids, exhibiting its characteristics from the other peas and beans. Regarding the health-functional content of GAB A, Turunoko and Tanbakuromame of Glycine max showed a high content. Similar to the classification depending on the external botanical structures and colors, the contents of the peas and beans, especially with the composition of the soluble sugars and amino acids, there are very distinct similarities and differences between difference species. Especially with Glycine max and Phaseolus vulgaris the difference was distinct. It has been confirmed that Sucrose and the amino acids related to umami could be used as ah \u27indicator\u27 to differentiate different species

Structural and functional insights into thermally stable cytochrome c' from a thermophile

Thermophilic Hydrogenophilus thermoluteolus cytochrome c0 (PHCP) exhibits higher thermal stability than a mesophilic counterpart, Allochromatium vinosum cytochrome c0 (AVCP), which has a homo-dimeric structure and ligand-binding ability. To understand the thermal stability mechanism and ligand-binding ability of the thermally stable PHCP protein, the crystal structure of PHCP was first determined. It formed a homo-dimeric structure, the main chain root mean square deviation (rmsd) value between PHCP and AVCP being 0.65 A ° . In the PHCP structure, six specific residues appeared to strengthen the heme-related and subunit–subunit interactions, which were not conserved in the AVCP structure. PHCP variants having altered subunit–subunit interactions were more severely destabilized than ones having altered heme-related interactions. The PHCP structure further revealed a ligand-binding channel and a penta-coordinated heme, as observed in the AVCP protein. A spectroscopic study clearly showed that some ligands were bound to the PHCP protein. It is concluded that the dimeric PHCP from the thermophile is effectively stabilized through heme-related and subunit–subunit interactions with conservation of the ligand-binding ability.This work was performed under the Cooperative Research Program of the “Network Joint Research Center for Materials and Devices”

Real-Time PCR Assay for the Diagnosis and Quantification of Co-infections by Diaporthe batatas and Diaporthe destruens in Sweet Potato

Foot rot disease caused by Diaporthe destruens (formerly Plenodomus destruens) has become a major concern for the production of sweet potato [Ipomoea batatas (L.) Lam.] in Japan. A related fungus Diaporthe batatas, which causes dry rot disease of sweet potato, is native and is widespread in fields in Japan. The similar characteristics of these two pathogens pose a challenge for conventional disease diagnosis. Currently, there are no effective molecular measures for identifying and distinguishing D. destruens and D. batatas. Here, we demonstrate a real-time PCR assay that distinguishes and quantifies D. batatas and D. destruens from co-infected sweet potato. The assay was performed with various simulated DNA combinations of D. batatas and D. destruens ranging from 1:1 to 1:100000. The assay was also used with the ratios of D. batatas: D. destruens: sweet potato DNA ranging from 1:1:1 to 1:1:100000. These assays produced a specific amplification product for each of the pathogens, and quantified the fungal biomass over the entire range tested without detecting false positives. The assay was validated by using infected sweet potato collected from various fields; it showed sufficient sensitivity and specificity to quantify and distinguish D. batatas and D. destruens from these field samples. Thus, our real-time PCR assay would be a useful tool for diagnosis of D. batatas and D. destruens and is expected to provide the foundation for the design of integrated disease management strategies for foot rot disease in sweet potato

ATM mediates pRBfunction to control DNMT1 protein stability and DNA methylation

The retinoblastoma tumor suppressor gene (RB) product has been implicated in epigenetic control of gene expression owing to its ability to physically bind to many chromatin modifiers. However, the biological and clinical significance of this activity was not well elucidated. To address this, we performed genetic and epigenetic analyses in an Rb-deficient mouse thyroid C cell tumor model. Here we report that the genetic interaction of Rb and ATM regulates DNMT1 protein stability and hence controls the DNA methylation status in the promoters of at least the Ink4a, Shc2, FoxO6, and Noggin genes. Furthermore, we demonstrate that inactivation of pRB promotes Tip60 (acetyltransferase)-dependent ATM activation; allows activated ATM to physically bind to DNMT1, forming a complex with Tip60 and UHRF1 (E3 ligase); and consequently accelerates DNMT1 ubiquitination driven by Tip60-dependent acetylation. Our results indicate that inactivation of the pRB pathway in coordination with aberration in the DNA damage response deregulates DNMT1 stability, leading to an abnormal DNA methylation pattern and malignant progression

Raft-based sphingomyelin interactions revealed by new fluorescent sphingomyelin analogs

Sphingomyelin (SM) has been proposed to form cholesterol-dependent raft domains and sphingolipid domains in the plasma membrane (PM). How SM contributes to the formation and function of these domains remains unknown, primarily because of the scarcity of suitable fluorescent SM analogs. We developed new fluorescent SM analogs by conjugating a hydrophilic fluorophore to the SM choline headgroup without eliminating its positive charge, via a hydrophilic nonaethylene glycol linker. The new analogs behaved similarly to the native SM in terms of their partitioning behaviors in artificial liquid order-disorder phase-separated membranes and detergent-resistant PM preparations. Single fluorescent molecule tracking in the live-cell PM revealed that they indirectly interact with each other in cholesterol- and sphingosine backbone–dependent manners, and that, for ∼10–50 ms, they undergo transient colocalization-codiffusion with a glycosylphosphatidylinositol (GPI)-anchored protein, CD59 (in monomers, transient-dimer rafts, and clusters), in CD59-oligomer size–, cholesterol-, and GPI anchoring–dependent manners. These results suggest that SM continually and rapidly exchanges between CD59-associated raft domains and the bulk PM

Effects of lifestyle education program for type 2 diabetes patients in clinics: study design of a cluster randomized trial

<p>Abstract</p> <p>Background</p> <p>The number of patients with type 2 diabetes is drastically increasing worldwide. It is a serious health problem in Japan as well. Lifestyle interventions can reduce progression from impaired glucose tolerance to type 2 diabetes, and glycemic control has been shown to improve postprandial plasma glucose levels. Moreover, several studies have suggested that continuous interventions (combined diet and exercise) can improve the plasma glucose level and reduce dosage of hypoglycemic agents.</p> <p>Although many interventional studies of lifestyle education for persons with diabetes in hospitals have been reported, only a few have been clinic-based studies employing an evidence-based lifestyle education program. This article describes the design of a cluster randomized controlled trial of the effectiveness of lifestyle education for patients with type 2 diabetes in clinics by registered dietitians.</p> <p>Methods/Design</p> <p>In Japan, general practitioners generally have their own medical clinics to provide medical care for outpatients in the community, including those with type 2 diabetes. With the collaboration of such general practitioners, the study patients were enrolled in the present study. Twenty general practitioners were randomly allocated to each provide patients for entry into either an intervention group (10) or a control group (10). In total, 200 participants will be included in the study. The intervention group will receive intensive education on lifestyle improvement related to type 2 diabetes by registered dietitians in clinics. Lifestyle education will be conducted several times during the study period. The control group will receive information on dietary intake and standard advice on glycemic control by registered dietitians. The primary endpoint is the change from the baseline value of HbA1c at 6 months. Data on health behavior and related issues will be gathered continuously over a 6-month period.</p> <p>Discussion</p> <p>This is the first study to evaluate lifestyle education in clinics by a cluster randomization trial in Japan. The proposed study will provide practical information about the usefulness of the intensive lifestyle improvement education program in primary care settings. The study was started in September 2007 and entry of subjects was completed in December 2010. Data on the effect evaluation will be available in 2011.</p> <p>Trial Registration</p> <p>UMIN000004049</p