Mutations in the gene encoding KIAA1199 protein,an inner-ear protein expressed in Deiters’ cells and the fibrocytes,as the cause of nonsyndromic hearing loss

We report three possibly disease-causing point mutations in one of the inner-ear-specific genes, KIAA1199. We identified an R187C mutation in one family, an R187H mutation in two unrelated families, and an H783Y mutation in one sporadic case of nonsyndromic hearing loss. In situ hybridization indicated that the murine homolog of KIAA1199 mRNA is expressed specifically in Deiters cells in the organ of Corti at postnatal day zero (Pn) P0 before the onset of hearing, but expression in those cells disappears by day P7. The signal of KIAA1199 was also observed in fibrocytes of the spiral ligament and the spiral limbus through to P21, when the murine cochlea matures. Thus, the gene product may be involved in uptake of potassium ions or trophic factors with a particular role in auditory development. Although the R187C and R187H mutations did not appear to affect subcellular localization of the gene product in vitro, the H783Y mutation did present an unusual cytoplasmic distribution pattern that could underlie the molecular mechanism of hearing impairment. Our data bring attention to a novel candidate for hearing loss and indicate that screening of mutations in inner-ear-specific genes is likely to be an efficient approach to finding genetic elements responsible for deafness.Nucleotide sequence data reported herein are available in the DDBJ/EMBL/GenBank databases; for details, see the electronic eatabase section of this article.     

Behavioral characteristics of rat lines selected for differential hypothermic responses to cholinergic or serotonergic agonists

The present review will describe the formation of two pharmacologically selected lines of rats, their behavioral phenotypes, their responses to select drugs, their possible neurochemical correlates, and their use to detect the therapeutic potential of antidepressant drugs. The Flinders Line rats were established at Flinders University in Australia by selectively breeding for differential responses to an anticholinesterase agent from outbred Sprague-Dawley (SD) rats; the Flinders Sensitive Line (FSL) rats were more sensitive to the hypothermic and behavioral suppressing effects of this agent than the Flinders Resistant Line (FRL) rats. The 8-OH-DPAT line rats were established at the University of North Carolina at Chapel Hill by selectively breeding for differential hypothermic responses to the 5-HT_1A receptor agonist, 8-OH-DPAT; the high DPAT sensitive (HDS) line rats were more sensitive to the hypothermic effects of 8-OH-DPAT than the low DPAT sensitive (LDS) line rats. Studies of these two pairs of lines have indicated that the FSL and HDS rats are both more susceptible to stress-induced behavioral disturbances. Their usefulness in detecting potential antidepressant drugs and the relationship between mood disorders and drug abuse will be discussed.     

Morphine-6beta-glucuronide modulates the expression of inducible nitric oxide synthase

The immunomodulatory effects of morphine are well established; however, suprisingly little is known about the immunomodulatory properties of the major metabolites of morphine. The present study tests the hypothesis that expression of inducible nitric oxide synthase (iNOS) is modulated by the administration of the morphine metabolite, morphine-6-glucuronide. The initial study using rats shows that morphine-6-glucuronide administration (0, 1.0, 3.163, 10 mg/kg s.c.) results in a pronounced reduction in lipopolysaccharide (LPS)-induced expression of iNOS (inducible nitricoxide synthease) in spleen, lung, and liver tissue as measured by western blotting. Morphine-6-glucuronide also produces a reduction in the level of plasma nitrite/nitrate, the more stable end-product of nitric oxide degradation. In a subsequent study, administration of the opioid receptor antagonist, naltrexone (0.1 mg/kg) prior to the injection of morphine-6-glucuronide (10 mg/kg) blocks the morphine-6-glucuronide induced reduction of iNOS expression and plasma nitrite/nitrite levels indicating that the effect is mediated via the opioid-receptor. This study provides the first evidence that morphine-6-glucuronide alters the expression of iNOS.     

Opa相互作用蛋白5在胰腺癌中的表达及其对PANC-1细胞增殖的影响

目的探索Opa相互作用蛋白5(OIP5)在胰腺癌中的表达及其对PANC-1细胞增殖的影响。方法通过数据库分析OIP5在胰腺癌组织及癌旁组织中的表达;用实时定量PCR(RT-qPCR)和蛋白印迹法(Western blot)分别检测人胰腺癌细胞系MIAPaCa-2、PANC-1、KP-3、BxPC-3细胞中OIP5 mRNA和蛋白表达;构建OIP5基因沉默质粒的慢病毒(pGCSIL-shOIP5)和对照质粒慢病毒(pGCSIL-shCtrl),分别感染PANC-1细胞,分为OIP5基因沉默组和shCtrl对照组,5 d后采用RT-qPCR和Western blot测定慢病毒敲低效率,流式细胞计量术检测细胞凋亡;OIP5基因沉默组和shCtrl对照组连续5 d进行MTT检测和细胞计数;OIP5基因沉默组和shCtrl对照组孵育10 d形成集落,Giemsa染色分别集落总数。结果胰腺癌中OIP5 mRNA表达显著高于正常胰腺组织(P<0.05),OIP5高表达患者的总存活率显著低于OIP5低表达患者(P<0.05),且其无病生存率也显著降低(P<0.05);OIP5在MIAPaCa-2、PANC-1和KP-3中表达较高,而在BxPC-3细胞系中的表达较低;MTT检测结果显示OIP5沉默在第4和第5天显著降低了PANC-1细胞的增殖速率(P<0.01);OIP5沉默后细胞集落数(平均为9个)显著低于shCtrl对照组中的数量(平均为40个)(P<0.01);OIP5沉默后PANC-1细胞凋亡比例为8.3%显著高于shCtrl的4.5%(P<0.01)。结论OIP5在胰腺癌细胞系中异常高表达,OIP5基因可调控胰腺癌PANC-1细胞的增殖、凋亡以及集落形成,提示OIP5可能在胰腺癌发病机制中作为癌基因发挥作用,从而为胰腺癌的靶向治疗提供了潜在的生物标志物。     

三磷酸肌醇在IL—2诱导的T细胞增殖分化中的信息传递作用

用不同浓度的IL-2刺激静息T淋巴细胞,其细胞内IP_3无明显改变,但ConA刺激则使IP_3增加45%。在IL-2依赖性T细胞,IL-2R表达率达83%,IL-2刺激时IP_3的变化依浓度不同而异,10u—50u/ml的IL-2使IP_3增高,以50u/ml最为显著,增加60%,但100u/ml IL-2及ConA不改变胞内IP_3的浓度。IL-2R封闭后的T淋巴细胞在IL-2刺激时IP_3的增加明显减弱。这些结果提示:IP_3作为细胞内的第二信使介导了IL-2诱导的T细胞的增殖反应,这种作用与IL-2的剂量及IL-2R表达有密切的关系。     

Distribution of Δ32 alelle of the CCR5 gene in the population of Poland

The chemokine receptor CCR5 constitutes a major co-receptor for the R5 strains of HIV-1, and a mutant allele of the CCR5 gene, especially in the homozygous form Δ32/Δ32, confers resistance against infection by the virus. The frequency of the Δ32 allele was determined in blood donors from 16 provinces, covering the entire territory of Poland. Among 861 individuals 182 (21.1%) were carriers of the mutated allele; 7 of them (0.8 %) were homozygotes Δ32/Δ32, and 175 (20.3%) were heterozygotes +/Δ32, resulting in a 10.9% frequency of the Δ32 allele. The highest frequencies of the mutated allele were found in the eastern and western provinces, and the lowest frequencies of the Δ32 allele were detected in the provinces in the center of the country. This pattern of distribution may reflect the migration of the population from the eastern territories of Poland to the western part of the country after World War II. Received: March 17, 2000 / Accepted: May 29, 2000     

Sequence of Echinochloa hoja blanca tenuivirus RNA-5

The sequence is presented of RNA-5 of Echinochloa hoja blanca tenuivirus, a second tenuivirus associated with rice cultivation in Latin America (after rice hoja blanca virus). The RNA is 1334 nucleotides long and contains in the complementary sense RNA a single long open reading frame. The deduced amino acid sequence of this open reading frame shows that it encodes a highly basic and hydrophilic 44 kD protein (pc5) with about 50% similarity to the pc5 protein of maize stripe virus (MStV). This and other features of the RNA are discussed.The GenBank accession number of the sequence reported in this paper is L47430.     

一个新的人B细胞活化抗原—5C5

用活化人B细胞株3D5细胞免疫小鼠和作为筛选的靶细胞,我们建立了产生单克隆抗体5C5的杂交瘤细胞株。此单抗识别的抗原5C5在25μg/ml anti-μ刺激的B细胞,于第10小时开始表达,亦即于G_1期开始表达。5C5细胞百分率随培养时间而增多。在PWM诱导下.外周血单一核细胞中5C5~ 细胞随培养时间而增加,至第3～4天达最高峰,然后减少,至第7天降至本底水平。5C5~ 细胞在不能为BCDF诱导分化至免疫球蛋白分泌细胞(ISC)的B细胞株3D5,Raji和Daudi阳性,但在能为BCDF诱导分化至ISC的CESS和SKW6细胞却不表达。这均表明5C5抗原表达于B细胞活化的早期和中期,但在B细胞终末分化阶段消失。在休止期B细胞、休止期T细胞、PHA激活的T细胞、单核细胞和中性粒细胞,以及在所检测的T细胞株和髓细胞株,5C5抗原均为阴性。~(125)I标记后用单抗5C5免疫沉淀提取的抗原,在还原与非还原条件下电泳,均只有分子量为52000的一条带,表明5C5是一个单链细胞表面蛋白。鉴于5C5抗原的分子量与文献中已报道的B细胞活化抗原分子量不同,以及5C5在细胞株表达的特点,它可能是一个新的人B细胞活化抗原。     

Familial partial trisomy 5p resulting from segregation of an insertional translocation

A case of duplication of segment p13-p15 of the short arm of chromosome 5 as the result of an insertional translocation in a mentally retarded girl with congenital anomalies is reported. Some of the apparently balanced carriers of the inverted insertion showed minor congenital anomalies.