The effect of high glucose on NO and O2- through endothelial GTPCH1 and NADPH oxidase

Although endothelial dysfunction deteriorates diabetic angiopathy, the mechanisms are obscure. We revealed that high glucose augmented eNOS through stimulation of eNOS mRNA in cultured BAECs. NO was decreased and O2- was increased simultaneously. NOS inhibitor, inhibited O2- release, so did NADPH oxidase inhibitor. The effects were synergistic. Both intracellular BH4 level and GTPCH1 activity were decreased by high glucose, in line with decrease of GTPCH1 mRNA. HMG-CoA reductase inhibitor, atorvastatin increased GTPCH1 mRNA and activity, and BH4 level. Conclusively, high glucose leads to eNOS dysfunction by inhibiting BH4 synthesis and atorvastatin stimulate BH4 synthesis directly, and it may work as atherogenic process.     

Area-based analyzing technique at cell array experiment using neuronal cell line

We propose a simple procedure for the identification and quantitative analysis of neurite outgrowth in neuronal cell lines that were uniformly differentiated. Upon stimulation most neuronal cell lines extend neurites in the differentiation process, resulting, according to our observation, in the increase of cell surface area. This increase is dependent on the length and the number of extended neurites. Furthermore, we use this method for the phenotype analysis of cell array experiments to perform large-scale functional evaluation of genes involved in the neurite outgrowth during neuronal differentiation.     

小鼠雌性生殖系统电穿孔与微泡增强超声波转EGFP基因的研究(简报)

非病毒载体转基因法,如注射裸DNA或脂质体转染,不产生细胞毒性,但除了肌肉组织外其他组织的转导效率均不高。电脉冲可使细胞膜产生临时的微孔允许一些分子通过,因此应用此方法可将药物或基因转入动物组织。电穿孔常用于培养的细胞转基因,理论上,低强度、长脉冲,或高强度、短脉冲有利于电转导,选择适合的参数是电转导的关键。本实验比较了不同电压和脉冲时间对小鼠卵巢在体转入绿色荧光蛋白基因的效果,确定了最适的电转导参数,为卵巢疾病的药物、基因治疗和研究卵泡发育中的基因调控提供了实验手段。超声波是临床常用的诊断方法,对人体无害…     

小鼠雌性生殖系统电穿孔与微泡增强超声波转EG即基因的研究（简报）

非病毒载体转基因法。如注射裸DNA或脂质体转染,不产生细胞毒性。但除了肌肉组织外其他组织的转导效率均不高。电脉冲可使细胞膜产生临时的微孔允许一些分子通过。因此应用此方法可将药物或基因转人动物组织。电穿孔常用于培养的细胞转基因,理论上,低强度、长脉冲。或高强度、短脉冲有利于电转导。选择适合的参数是电转导的关键。本实验比较了不同电压和脉冲时间对小鼠卵巢在体转入绿色荧光蛋白基因的效果．确定了最适的电转导参数。为卵巢疾病的药物、基因治疗和研究卵泡发育中的基因调控提供了实验手段。     

Apoptotic cells activate NKT cells through T cell Ig-like mucin-like-1 resulting in airway hyperreactivity

T cell Ig-like mucin-like-1 (TIM-1) is an important asthma susceptibility gene, but the immunological mechanisms by which TIM-1 functions remain uncertain. TIM-1 is also a receptor for phosphatidylserine (PtdSer), an important marker of cells undergoing programmed cell death, or apoptosis. We now demonstrate that NKT cells constitutively express TIM-1 and become activated by apoptotic cells expressing PtdSer. TIM-1 recognition of PtdSer induced NKT cell activation, proliferation, and cytokine production. Moreover, the induction of apoptosis in airway epithelial cells activated pulmonary NKT cells and unexpectedly resulted in airway hyperreactivity, a cardinal feature of asthma, in an NKT cell-dependent and TIM-1-dependent fashion. These results suggest that TIM-1 serves as a pattern recognition receptor on NKT cells that senses PtdSer on apoptotic cells as a damage-associated molecular pattern. Furthermore, these results provide evidence for a novel innate pathway that results in airway hyperreactivity and may help to explain how TIM-1 and NKT cells regulate asthma.     

Determination of antifungal activities in serum samples from mice treated with different antifungal drugs allows detection of an active metabolite of itraconazole

To establish an in vitro method of predicting in vivo efficacy of antifungal drugs against Candida albicans and Aspergillus fumigatus, the antifungal activities of fluconazole, itraconazole, and amphotericin B were determined in mouse serum. The minimum inhibitory concentration (MIC) of each drug was measured using mouse serum as a diluent. For C. albicans, the assay endpoint of azoles was defined as inhibition of mycelial extension (mMIC) and for A. fumigatus, as no growth (MIC). The MICs of amphotericin B for both pathogens were defined as the MIC at which no mycelial growth occurred. Serum MIC or mMIC determinations were then used to estimate the concentration of the drugs in serum of mice treated with antifungal drugs by multiplying the antifungal titer of the serum samples by the serum (m)MIC. The serum drug concentrations were also determined by HPLC. The serum concentrations estimated microbiologically showed good agreement with those determined by HPLC, except for itraconazole. Analysis of the serum samples from itraconazole-treated mice by a sensitive bioautography revealed the presence of additional spots, not seen in control samples of itraconazole. The bioautography assay demonstrated that the additional material detected in serum from mice treated with itraconazole was an active metabolite of itraconazole. The data showed that the apparent reduction in the itraconazole serum concentration as determined by HPLC was the result of the formation of an active metabolite, and that the use of a microbiological method to measure serum concentrations of drugs can provide a method for prediction of in vivo efficacy of antifungal drugs.     

Effects of chlorogenic acid and its metabolites on spontaneous locomotor activity in mice

Chlorogenic acid possessed a weak caffeine-like psychostimulant property when assessed for its effect on spontaneous locomotor activity in mice. In the evaluation of the effects for the major metabolites of chlorogenic acid which were detected upon incubation with rat feces and/or excreted in urine after oral administration to rats, caffeic and m-coumaric acids were found to be the principal active metabolites, while the others contributed little to this caffeine-like psychostimulant activity.     

Horizontal distribution of Thetys vagina Tilesius (Tunicata, Thaliacea) in the Japan Sea during spring 2004

The occurrence of the salp Thetys vagina was observed in theJapan Sea during spring 2004. Catches up to 187 kg wet weight(WW) per 2.18 x 10⁵ m³ (equal to 0.9 g WW m^–3) were collectedwith 10-m diameter surface-water otter trawl nets. The horizontaldistribution indicated that the high biomass was related tothe area with high chlorophyll a (Chl a) concentration, whichwas located around the subarctic front with the warm TsushimaCurrent. Five prey taxa were identified from the gut contentsof individuals from the high Chl a area. The diatom Coscinodiscusspp. (13–55 µm in diameter) dominated numerically.Another significant prey was the large diatom Coscinodiscuswailesii (219–313 µm) that is an indicator of thespring bloom in this area. The mass occurrence of T. vaginathus appears related to phytoplankton availability, though themechanisms remain uncertain.