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311.
312.
Clusterin has been known to play important roles in cell-cell and/or cell-substratum interactions. Recently we reported the transient expression of clusterin in pancreatic endocrine cells during the early developmental stages and suggested a role in aggregating the endocrine cells for islet formation. In the present study, we have investigated the involvement of clusterin in cell-substratum interaction by the inhibition of clusterin synthesis using antisense oligonucleotide. The expression of clusterin was transiently increased as early as 2–8 h after plating the ASC-17D Sertoli cells to the culture flask, which was the period of cell attachment. In addition, up-regulation of clusterin mRNA was so much greater when the Sertoli cells were plated on the petri dish for the bacterial culture instead of in a animal cell culture flask that therefore, the cells failed to attach to it. These findings suggested that interruption of cell to plate substratum interaction might lead to over-expression of clusterin from Sertoli cells to induce cell to cell aggregation or, perhaps, to re-establish attachment with the substratum. Transfection of ASC-17D Sertoli cells with a 20-base antisense oligonucleotide against clusterin mRNA resulted in extracellular release of LDH and DNA fragmentation. Sertoli cell death by antisense oligonucleotide of clusterin was sequence specific and dose dependent. Treatment of antisense oligonucleotide induced a marked reduction of synthesis for clusterin protein, but not for clusterin mRNA expression, suggesting the translational suppression of clusterin by antisense oligonucleotide. Further, microscopic observation showed that more noticeable cell death was induced by treating the antisense prior to plating the cells than by treating after cell attachment to the plate. From these results, we speculate that down-regulation of clusterin expression in the anchorage-dependent Sertoli cells prevents them from attaching to the plate, and therefore induces cell death. 相似文献
313.
Cytoprotective effect of arginine deiminase on taxol-induced apoptosis in DU145 human prostate cancer cells 总被引:3,自引:0,他引:3
Kang SW Kang H Park IS Choi SH Shin KH Chun YS Chun BG Min BH 《Molecules and cells》2000,10(3):331-337
We purified and partially sequenced a cytostatic protein from the ASC-17D Sertoli cell-conditioned media (rSCCM) showing a molecular weight of 90 kDa with homodimeric composition. N-terminal amino acid analysis revealed that the protein was homologous to the arginine deiminase (ADI) of Mycoplasma arginini. We found ADI enzyme activity in rSCCM and the abolishment of the growth inhibitory effect by the supplement of L-arginine. Thus, we confirmed that the cytostatic activity in rSCCM was due to the depletion of extracellular L-arginine by ADI. Apparent increase of cell death or DNA fragmentation was not observed in DU145 cells cultured in the presence of ADI. Incubation of DU145 cancer cells with taxol resulted in a marked DNA fragmentation, whereas pretreatment with ADI or cycloheximide protected the cells from taxol-induced apoptosis. Preincubation of the cells with ADI inhibited S35-methionine incorporation into protein synthesis in a dose dependent manner. These data suggest that ADI-induced arginine depletion may inhibit protein synthesis, and result in the protection of apoptotic cell death that requires new protein synthesis. 相似文献
314.
On the mechanisms of T cell silencing by IL-10 DNA: direct and indirect inhibition of T cell functions 总被引:2,自引:0,他引:2
Previously we reported that mucosal IL-10 DNA administration resulted in long-term suppression of virus-induced inflammatory responses by silencing Th1-type CD4+ T cell functions. However, the mechanism by which IL-10 silences the activity of CD4+ T cells was not clear. The present report has shown that mucosal IL-10 DNA administration led to the reduction of reactivity of T cells following TCR stimulation. IL-10 DNA also downregulated APC functions to stimulate T cells but the effect was temporary. Bystander suppression, including that of IL-10 producing regulatory cells, appeared not to be directly involved in the inhibition of T cell reactivity because both anti-IL-10 and anti-IL-10R could not block the suppression of T cell functions. This silenced state could be maintained following adoptive transfer to untreated animals. The nature of the silencing appears to be a reversible anergic state since Ag stimulation in the presence of exogenous IL-2 restored T cell reactivity. Furthermore, IL-10-induced silenced T cells could be induced in vitro by culturing the T cells with rIL-10 in the presence or the absence of antigen stimulation. This state persisted in the absence of rIL-10 and persisted for at least 3 days. A more notable effect, however, was observed when the T cells were incubated with IL-10 in the presence of APC and Ag. These results indicate that IL-10 induced a long-term silenced state in T cells by direct and indirect inhibition of T cell functions. 相似文献
315.
Chae Young-Am Hee-Suk Yu Ji-Sook Song Hee-Kyung Chun Sang-Un Park 《Biotechnology letters》2000,22(19):1527-1530
The transformed root culture of Polygonum tinctorium Lour. was established by infecting leaf explants with Agrobacterium rhizogenes A4. These cultures were examined for their growth and indigo content under various culture conditions. Among the four different culture media tested, SH medium showed the highest yield for root growth (28 mg dry wt/30 ml) and indigo production (152 g/dry wt). In SH medium, 30 g sucrose l–1, 2500 mg KNO3 l–1, 300 mg NH4H2PO4 l–1 were the best conditions for indigo production at pH 5.7. The production of indigo in hairy roots slightly increased with the addition of 200 mg chitosan l–1 (186 g/dry wt) and 20 U pectinase l–1 (181 g/dry wt). 相似文献
316.
Ataxia-telangiectasia: identification and detection of founder-effect mutations in the ATM gene in ethnic populations. 总被引:5,自引:0,他引:5
M Telatar S Teraoka Z Wang H H Chun T Liang S Castellvi-Bel N Udar A L Borresen-Dale L Chessa E Bernatowska-Matuszkiewicz O Porras M Watanabe A Junker P Concannon R A Gatti 《American journal of human genetics》1998,62(1):86-97
To facilitate the evaluation of ATM heterozygotes for susceptibility to other diseases, such as breast cancer, we have attempted to define the most common mutations and their frequencies in ataxia-telangiectasia (A-T) homozygotes from 10 ethnic populations. Both genomic mutations and their effects on cDNA were characterized. Protein-truncation testing of the entire ATM cDNA detected 92 (66%) truncating mutations in 140 mutant alleles screened. The haplotyping of patients with identical mutations indicates that almost all of these represent common ancestry and that very few spontaneously recurring ATM mutations exist. Assays requiring minimal amounts of genomic DNA were designed to allow rapid screening for common ethnic mutations. These rapid assays detected mutations in 76% of Costa Rican patients (3), 50% of Norwegian patients (1), 25% of Polish patients (4), and 14% of Italian patients (1), as well as in patients of Amish/Mennonite and Irish English backgrounds. Additional mutations were observed in Japanese, Utah Mormon, and African American patients. These assays should facilitate screening for A-T heterozygotes in the populations studied. 相似文献
317.
青蒿转杜松烯合成酶基因发根系的培养 总被引:10,自引:2,他引:8
将已克隆的棉花杜松烯合成酶的cDNA(cadC14)插入到植物表达载体pBI121中,构建含CaMV35S启动子驱动下的杜松烯合成酶基因的植物表达载体pBIC14。用含pBIC14质粒的发根农杆菌(Agrobacteriumrhizogenes)15834感染青蒿(ArtemisiaannuaL.)叶片并诱导发根,共建立121个生长迅速的发根系。经浓度为20mg/L的Kan筛选,获得12个抗Kan阳性根系。PCR和Southernbloting分析表明,外源杜松烯合成酶基因已整合到青蒿基因组中,其转基因频率为3%。RTPCR分析表明,外源杜松烯合成酶基因在C37根系中,在转录水平上已有表达。 相似文献
318.
青蒿毛状根合成青蒿素的培养条件研究 总被引:14,自引:0,他引:14
对影响青蒿(ArtemisiaannuaL.)毛状根生长及青蒿素合成的培养条件进行了研究,确定最适的培养条件为:初始pH5.8~6.0,摇瓶转速130~150r/min,摇瓶装液量体积分数为25%,光照周期为16h/d,温度为30℃。在此条件下,经过25d培养获得青蒿素产量为223.3mg/L。 相似文献
319.
Chun Fang Shen Serge R. Guiot Sonia Thiboutot Guy Ampleman Jalal Hawari 《Biodegradation》1998,8(5):339-347
Microcosm tests simulating bioslurry reactors with 40% soilcontent, containing high concentrations of TNT and/or RDX,and spiked with either [14C]-TNT or[14C]-RDX were conducted to investigate the fate ofexplosives and their metabolites in bioslurry treatment processes.RDX is recalcitrant to indigenous microorganisms in soil andactivated sludge under aerobic conditions. However, soilindigenous microorganisms alonewere able to mineralize 15% of RDX to CO2 underanaerobic condition, and supplementation of municipal anaerobicsludge as an exogenous source of microorganismssignificantly enhanced the RDX mineralization to 60%. RDXmineralizing activity of microorganisms in soil and sludge wassignificantly inhibited by the presence of TNT. TNTmineralization was poor (< 2%) and was not markedlyimproved by the supplement ofaerobic or anaerobic sludge. Partitioning studies of[14C]-TNT in the microcosmsrevealed that the removal of TNTduring the bioslurry process was due mainly to thetransformation of TNT and irreversiblebinding of TNT metabolites onto soil matrix. In the case ofRDX under anaerobic conditions,a significant portion (35%) of original radioactivity wasalso incorporated into the biomass andbound to the soil matrix. 相似文献
320.
马立克氏病病毒(Marek’sdiseasevirus,MDV)是一种能诱导鸡淋巴组织增生或淋巴肿瘤的细胞结合性疱疹病毒,由此而引起的马立克氏病(Marek’sdisease,MD)也是迄今为止唯一可以利用疫苗进行有效控制的肿瘤性疾病。鉴于此,作为研... 相似文献