Quantitative bioimaging analysis of gonads in olvas-GFP/ST-II YI Medaka (transgenic Oryzias latipes) exposed to ethinylestradiol

This study investigates the adverse and persistent effects of ethinylestradiol (EE2) on mature gonads of transgenic olvas-GFPIST II-YI medaka (Oryzias latipes). The measurement of gonadal size calculating the GFP-fluorescent area was used as a technique that enabled monitoring gonads in living specimens by GFP fluorescence. First, mature medaka were exposed to EE2 (47.8-522 ng/L) for 4 weeks. The gonads showed a significant reduction of the GFP-fluorescent area and Gonadosomatic Index in males exposed to EE2 at >216 ng/L and females exposed at 522 ng/L. Histologically, males at all treatments exhibited testis-ova and additionally, high connective tissue prevalence at > or =216 ng/L. Next, mature male medaka were exposed to EE2 (43.7-473 ng/L) for 3 weeks and allowed to depurate for 6 weeks, to investigate persistent effects of EE2. Continuous gonad observation showed that GFP began to decline 3 weeks after initial exposure to > or =215 ng/L. After depuration, the gonad's fluorescent areas gradually recovered, with no statistical difference at the end of the depuration period; normal spermatogenesis was present in these individuals. Alterations in GFP fluorescence clearly indicate the condition of the gonad in transgenic medaka and this strain showed a facilitated screening fish model to detect the adverse effects on the gonad by estrogenic chemicals.     

SEM observation of wet biological specimens pretreated with room-temperature ionic liquid

A facile pretreatment process for SEM: The use of room temperature ionic liquids (RTILs) provides an interesting method for SEM of biological specimens. We used a novel and concise method of pretreatment, excluding fixation or Au sputtering steps. Fine and smooth-textured SEM images of a wide variety of biological specimens treated in this way were observed without artefacts.     

Modulation of interferon-γ synthesis by the effects of lignin-like enzymatically polymerized polyphenols on antigen-presenting cell activation and the subsequent cell-to-cell interactions

Lignin-like polymerized polyphenols strongly activate lymphocytes and induce cytokine synthesis. We aimed to characterise the mechanisms of action of polymerized polyphenols on immunomodulating functions. We compared the reactivity of leukocytes from various organs to that of polymerized polyphenols. Splenocytes and resident peritoneal cavity cells (PCCs) responded to polymerized polyphenols and released several cytokines, whereas thymocytes and bone-marrow cells showed no response. Next, we eliminated antigen-presenting cells (APCs) from splenocytes to study their involvement in cytokine synthesis. We found that APC-negative splenocytes showed significantly reduced cytokine production induced by polymerized polyphenols. Additionally, adequate interferon-γ (IFN-γ) induction by polymerized polyphenols was mediated by the coexistence of APCs and T cells because the addition of T cells to PCCs increased IFN-γ production. Furthermore, inhibition of the T cell-APC interaction using neutralising antibodies significantly decreased cytokine production. Thus, cytokine induction by polymerized polyphenols was mediated by the interaction between APCs and T cells.     

Effect of Ganglioside GM3 Synthase Gene Knockout on the Glycoprotein N‐Glycan Profile of Mouse Embryonic Fibroblast

The structural and clinical significance of cellular glycoproteins and glycosphingolipids (GSLs) are often separately discussed. Considering the biosynthetic pathway of glycoconjugates, glycans of cell‐surface glycoproteins and GSLs might partially share functions in maintaining cellular homeostatis. The purpose of this study is to establish a general and comprehensive glycomics protocol for cellular GSLs and N‐glycans of glycoproteins. To test the feasibility of a glycoblotting‐based protocol, whole glycans released both from GSLs and glycoproteins were profiled concurrently by using GM3 synthase‐deficient mouse embryonic fibroblast GM3(?/?). GM3(?/?) cells did not synthesize GM3 or any downstream product of GM3 synthase. Instead, expression levels of o‐series gangliosides involving GM1‐b and GD1‐α increased dramatically, whereas a‐/b‐series gangliosides were predominantly detected in wild‐type (WT) cells. We also discovered that glycoprotein N‐glycan profiles of GM3(?/?) cells are significantly altered as compared to WT cells, although GM3 synthase is responsible only for GSLs synthesis and is not associated with glycoprotein N‐glycan biosynthesis. The present approach allows for high‐throughput profiling of cellular glycomes enriched by different classes of glycoconjugates, and our results demonstrated that gene knockout of the enzymes responsible for GSL biosynthesis significantly influences the N‐glycans of glycoproteins.     

α,ω-二元羧酸单酯在铑系双组份催化剂上加氢制ω-羟基羧酸酯

在小型高压反应釜中研究了铑系催化剂对于α,ω－ 二元羧酸单酯加氢生成ω－ 羟基羧酸酯的催化性能。结果表明,Ｒｈ － Ｒｅ 以及Ｒｈ － Ｍｏ 双组份催化剂具有良好的催化加氢性能;α,ω－ 十五烷二酸单甲酯在Ｒｈ － Ｒｅ 和Ｒｈ － Ｍｏ双组份催化剂上加氢生成ω－ 羟基十五烷酸甲酯的收率分别达到８７－５ ％ 和８４－７ ％ ;其它α,ω－ 二元羧酸单酯在Ｒｈ －Ｍｏ 双组份催化剂上加氢生成相应的ω－ 羟基羧酸酯的收率也达到８０－９ ％ ～９１－３ ％ 。反应温度对α,ω－ 二元羧酸单酯加氢生成ω－ 羟基羧酸酯的收率有较大的影响,乙二醇二甲醚作为溶剂有利于目的产物的生成。     

Cytoprotective effects of geraniin against peroxynitrite- and peroxyl radical-induced cell death via free radical scavenging activity

The role of natural antioxidants in combating the deleterious effects of free radicals has received much attention. In the present study, the cytoprotective effects and the free radical scavenging activity of geraniin, a hydrolysable ellagitannin from Nephelium lappaceum rind, were evaluated by using various approaches. Addition of geraniin to the culture media resulted in a profound cytoprotective effect against damages induced by the peroxynitrite generator 3-morpholinosydnonimine (SIN-1), and the peroxyl radical generator 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH). Geraniin exhibits more potent cytoprotective activity than that of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox). Geraniin exhibited potent antioxidant activity against reactive species, such as nitric oxide, superoxide anion, and chemically synthesized peroxynitrite. Kinetic analysis of reactivity against peroxyl radicals generated by AAPH revealed that geraniin possesses potent reactivity against peroxyl radicals with higher stoichiometric number than Trolox. The cytoprotective effect of geraniin was only observed when geraniin and these toxic compounds were co-existing, suggesting that geraninn exhibits cytoprotective effects via free radical scavenging activity in the extracellular fluid.     

An application of the 2-nitrobenzenesulfenyl method to proteomic profiling of human colorectal carcinoma: A novel approach for biomarker discovery

In the development of novel biomarkers, the proteomic approach is advantageous because using it the cancer-associated proteins can be directly identified. We previously developed a 2-nitrobenzenesulfenyl (NBS) method to improve quantitative proteome analysis. Here, we applied this method to proteomic profiling of colorectal carcinoma (CRC) to identify novel proteins with altered expression in CRC. Each pair of tumor and normal tissue specimens from 12 CRC patients was analyzed, and approximately 5000 NBS-labeled paired peaks were quantified. Peaks with altered signal intensities (>1.5-fold) and occurring frequently in the samples (>70%) were selected, and 128 proteins were identified by MS/MS analyses as differentially expressed proteins in CRC tissues. Many proteins were newly revealed to be CRC related; 30 were reported in earlier studies of CRC. Six proteins that were up-regulated in CRC (ZYX, RAN, RCN1, AHCY, LGALS1, and VIM) were further characterized and validated by Western blot and immunohistochemistry. All six were found to be CRC-localized, either in cancer cells or in stroma cells near the cancer cells. These results indicate that the proteins identified in this study are novel candidates for CRC markers, and that the NBS method is useful in proteome mining to discover novel biomarkers.     

Exoskeletal meal assistance system (EMAS II) for patients with progressive muscular disease

Patients with muscle weakness such as muscular dystrophy usually need someone’s assistance in their daily activities. In order to reduce the caregiver burden and to improve quality of life (QOL) of the patients, various robotic technologies have been developed. This paper presents an exoskeletal assistance system EMAS II for the patients, which assists the upper extremity for the purpose of daily activities such as eating, writing, or other desk works. The EMAS II assists four DOF; shoulder flexion-extension, shoulder abduction-adduction, shoulder medial-lateral rotation, and elbow flexion-extension. The EMAS II has three kinds of user interfaces which are operated by residual functions of the patients, because it is important for patients’ health and initiative to use the residual functions. In order to control the four DOFs exoskeleton system using the interfaces with less DOF, the EMAS II simulates upper limb motion patterns of healthy people. The patterns are modeled by extracting correlations between the height of the wrist joint and that of the elbow joint. Therefore, users have only to control the position of their wrist joint to do tasks at a table. Through an experiment with a healthy subject, the feasibility of meal assistance by the EMAS II was confirmed. Furthermore, the system was applied to a spinal muscular atrophy patient in a clinical trial to check the usability. The experimental results indicated that the EMAS II could support the patient’s upper extremity to do tasks at a table.     

Detection of Verotoxigenic Escherichia coli O157 and O26 in food by plating methods and LAMP method: a collaborative study

In order to establish a rapid and sensitive method for the detection of Verotoxigenic Escherichia coli O157 and O26, a collaborative study was conducted focusing on a comparison of the efficiency of loop-mediated amplification (LAMP) assay targeting the Verocytotoxin (also called Shiga toxin) gene, utilizing a direct plating method and a plating method with immunomagnetic separation (IMS-plating method) using various agar media. In combination with enrichment with the modified EC supplemented with novobiocin, E. coli O157 was detected in most samples of ground beef and alfalfa sprouts by LAMP assay, the direct plating method and the IMS-plating method. E. coli O26 was detected in approximately 100% of the food samples by LAMP assay. However, the IMS-plating and direct plating methods recovered 80 and 50% in ground beef samples, respectively. As a result, it was demonstrated the LAMP assay is superior to the IMS-plating method. Based on these results, it appears LAMP assay is effective as a screening assay to detect E. coli O157 and O26 from positive samples.     

Ultra light-weight and high-resolution X-ray mirrors using DRIE and X-ray LIGA techniques for space X-ray telescopes

We are developing novel ultra light-weight and high-resolution X-ray micro pore optics for space X-ray telescopes. In our method, curvilinear micro pore structures are firstly fabricated by silicon deep reactive ion etching (DRIE) or X-ray LIGA processes. Secondly, side walls of the micro structures are smoothed by magnetic field assisted finishing and/or hydrogen annealing techniques for high reflectivity mirrors. Thirdly, to focus parallel X-ray lights from astronomical objects, these structures are elastically or plastically bent into a spherical shape. Fourthly, the bent structures are stacked to form a multi-stage X-ray telescope. In this paper, we report on fabrication and X-ray reflection tests of silicon and nickel X-ray mirrors using the DRIE and LIGA processes, respectively. For the first time, X-ray reflections were confirmed on both of the mirrors. Estimated rms roughnesses were 5 nm and 3 nm for the silicon and nickel mirrors, respectively.