Estimates of sperm sex chromosome disomy and diploidy rates in a 47,XXY/46,XY mosaic Klinefelter patient

A 47,XXY/46,XY male was investigated for the incidence of aneuploidy in sperm sex chromosomes using a three-colour X/Y/18 fluorescence in situ hybridisation (FISH) protocol. A total of 1701 sperm nuclei were analysed. The ratio of X-bearing to Y-bearing sperm did not differ from the expected 1 : 1 ratio although there were more 23,Y sperm than 23,X sperm (844 vs 795). There was a significantly increased proportion of disomy XY and XX sperm compared with normal controls (0.41% vs 0.10%, P < 0.001 and 0.29% vs 0.04%, P < 0.01). However, the incidence of YY sperm was similar to the controls (0.06% vs 0.02%). The diploidy rate was also significantly increased (1.7% vs 0.13%, P < 0.0001), as was disomy 18 (0.71% vs 0.01%) and 25,XXY (0.47% vs 0%). The results support the hypothesis that some 47,XXY cells are able to undergo meiosis and produce mature spermatozoa. Patients with mosaic Klinefelter syndrome with severe oligozoospermia have significantly elevated incidences of disomy XY and XX sperm and may be at a slightly increased risk of producing 47,XXX and 47,XXY offspring. Additionally, they may be at risk of producing offspring with autosomal trisomies. Hence, patients with Klinefelter mosaicism scheduled for intracytoplasmic sperm injection intervention should first undergo FISH analysis of their sperm to determine their risk. Received: 16 November 1998 / Accepted: 16 February 1999     

Different metabolic responses to central and peripheral injection of enterostatin

Enterostatin, a pentapeptide cleaved from procolipase, suppresses fat intake after peripheral and central administration. Chronic treatment of rats with enterostatin decreases body weight and body fat. The effect was greater than could be accounted by the reduction in food intake alone. Hence, we have investigated the effect of enterostatin on energy metabolism. Male Sprague-Dawley rats adapted to a high-fat diet were implanted with lateral cerebral ventricular or amygdala cannulas. The metabolic effects were determined by indirect calorimetry. After habituation to the test cages, fasted rats were injected with either saline vehicle or enterostatin given either intraperitoneally (100 nmol) or intracerebroventricularly (1 nmol) or into specific brain regions [amygdala (0.01 nmol) or paraventricular nucleus (PVN) (0.1 nmol)]. Respiratory quotient (RQ) and energy expenditure were monitored over 2 h. Intraperitoneal enterostatin reduced RQ (saline: 0.81 +/- 0.02 vs. enterostatin: 0.76 +/- 0.01) and increased energy expenditure by 44%. Intracerebroventricular enterostatin increased the energy expenditure without any effects on RQ, whereas PVN enterostatin increased metabolic rate, while preventing the increase in RQ observed in the control animals. In contrast, neither RQ nor energy expenditure was altered after enterostatin was injected into the amygdala. Enterostatin activated AMP-activated protein kinase in primary cultures of human myocytes in a dose- and time-dependent manner and increased the rate of fatty acid beta-oxidation. These findings suggest that enterostatin regulates energy expenditure and substrate partitioning through both peripheral and central effects.     

Id1 interacts and stabilizes the Epstein-Barr virus latent membrane protein 1 (LMP1) in nasopharyngeal epithelial cells

The EBV-encoded latent membrane protein 1 (LMP1) functions as a constitutive active form of tumor necrosis factor receptor (TNFR) and activates multiple downstream signaling pathways similar to CD40 signaling in a ligand-independent manner. LMP1 expression in EBV-infected cells has been postulated to play an important role in pathogenesis of nasopharyngeal carcinoma. However, variable levels of LMP1 expression were detected in nasopharyngeal carcinoma. At present, the regulation of LMP1 levels in nasopharyngeal carcinoma is poorly understood. Here we show that LMP1 mRNAs are transcribed in an EBV-positive nasopharyngeal carcinoma (NPC) cell line (C666-1) and other EBV-negative nasopharyngeal carcinoma cells stably re-infected with EBV. The protein levels of LMP1 could readily be detected after incubation with proteasome inhibitor, MG132 suggesting that LMP1 protein is rapidly degraded via proteasome-mediated proteolysis. Interestingly, we observed that Id1 overexpression could stabilize LMP1 protein in EBV-infected cells. In contrary, Id1 knockdown significantly reduced LMP1 levels in cells. Co-immunoprecipitation studies revealed that Id1 interacts with LMP1 by binding to the CTAR1 domain of LMP1. N-terminal region of Id1 is required for the interaction with LMP1. Furthermore, binding of Id1 to LMP1 suppressed polyubiquitination of LMP1 and may be involved in stabilization of LMP1 in EBV-infected nasopharyngeal epithelial cells.     

Absence of Intestinal PPARγ Aggravates Acute Infectious Colitis in Mice through a Lipocalin-2–Dependent Pathway

To be able to colonize its host, invading Salmonella enterica serovar Typhimurium must disrupt and severely affect host-microbiome homeostasis. Here we report that S. Typhimurium induces acute infectious colitis by inhibiting peroxisome proliferator-activated receptor gamma (PPARγ) expression in intestinal epithelial cells. Interestingly, this PPARγ down-regulation by S. Typhimurium is independent of TLR-4 signaling but triggers a marked elevation of host innate immune response genes, including that encoding the antimicrobial peptide lipocalin-2 (Lcn2). Accumulation of Lcn2 stabilizes the metalloproteinase MMP-9 via extracellular binding, which further aggravates the colitis. Remarkably, when exposed to S. Typhimurium, Lcn2-null mice exhibited a drastic reduction of the colitis and remained protected even at later stages of infection. Our data suggest a mechanism in which S. Typhimurium hijacks the control of host immune response genes such as those encoding PPARγ and Lcn2 to acquire residence in a host, which by evolution has established a symbiotic relation with its microbiome community to prevent pathogen invasion.     

Mutations in QARS,Encoding Glutaminyl-tRNA Synthetase,Cause Progressive Microcephaly,Cerebral-Cerebellar Atrophy,and Intractable Seizures

Progressive microcephaly is a heterogeneous condition with causes including mutations in genes encoding regulators of neuronal survival. Here, we report the identification of mutations in QARS (encoding glutaminyl-tRNA synthetase [QARS]) as the causative variants in two unrelated families affected by progressive microcephaly, severe seizures in infancy, atrophy of the cerebral cortex and cerebellar vermis, and mild atrophy of the cerebellar hemispheres. Whole-exome sequencing of individuals from each family independently identified compound-heterozygous mutations in QARS as the only candidate causative variants. QARS was highly expressed in the developing fetal human cerebral cortex in many cell types. The four QARS mutations altered highly conserved amino acids, and the aminoacylation activity of QARS was significantly impaired in mutant cell lines. Variants p.Gly45Val and p.Tyr57His were located in the N-terminal domain required for QARS interaction with proteins in the multisynthetase complex and potentially with glutamine tRNA, and recombinant QARS proteins bearing either substitution showed an over 10-fold reduction in aminoacylation activity. Conversely, variants p.Arg403Trp and p.Arg515Trp, each occurring in a different family, were located in the catalytic core and completely disrupted QARS aminoacylation activity in vitro. Furthermore, p.Arg403Trp and p.Arg515Trp rendered QARS less soluble, and p.Arg403Trp disrupted QARS-RARS (arginyl-tRNA synthetase 1) interaction. In zebrafish, homozygous qars loss of function caused decreased brain and eye size and extensive cell death in the brain. Our results highlight the importance of QARS during brain development and that epilepsy due to impairment of QARS activity is unusually severe in comparison to other aminoacyl-tRNA synthetase disorders.     

陕甘宁地区降水稳定同位素特征及水汽来源

依据全球大气降水稳定同位素观测网络(GNIP)和已有研究中陕甘宁地区的大气降水氢氧稳定同位素资料,并结合相关气象数据,分析了陕甘宁地区大气降水氧稳定同位素的时空分布特征及其影响因子,并建立了大气降水线方程.结果表明:3省大气降水线的斜率、截距由小到大依次为:甘肃、陕西、宁夏,且均小于全球大气降水线方程的斜率、截距,表明3省的降水过程受蒸发作用影响程度沿宁夏、陕西、甘肃增强;陕甘宁地区大气降水中δ18O值在时间变化上,表现为夏秋季节富集、冬春季节贫化,从空间分布来看,由西北至东南,加权平均δ18O值呈减小趋势;3省降水中δ18O温度效应显著,但不存在降水量效应,这体现了中高纬度大陆性气候的特征;高程效应的定量关系为-0.12‰·(100 m)^-1,纬度效应更显著(纬度每增加1°,降水中δ18O相应贫化0.27‰);采用HYSPLIT模型对各站点的水汽来源进行追踪,气团聚类轨迹表明,夏半年主要有来自孟加拉湾的水汽、东南季风水汽和西风带水汽,冬半年以西风带水汽为主.     

The characteristics and enzyme activities of 4-chlorophenol biodegradation by Fusarium sp

The effects of pH, temperature and sucrose addition on biodegradative capacity of Fusarium sp. HJ01 for 4-chlorophenol (4-CP) were examined, the property of dioxygenases produced by Fusarium sp. HJ01 during 4-CP degradation was investigated. The results show that Fusarium sp. HJ01 has a high capacity on degrading 4-CP in solution. The optimum values of pH, sucrose concentration and temperature are pH 7,1 g/L and 30°C, respectively. The strain can produce chlorocatechol 1,2-dioxygenase (CC12O) and chlorocatechol 2,3-dioxygenase (CC23O), which show the highest activities when 4-CP is used as the sole carbon source and energy, and the optimal values of pH and temperature are pH 7 and 50°C for CC12O as well as pH 8 and 60°C for CC23O. The kinetics of enzyme-catalyzed reactions accord with the Michaelis-Menten equation. To our knowledge, this is the first study on biodegradation of 4-CP by Fusarium sp. HJ01.     

红火蚁蚁丘大小与诱集工蚁数量的关系

采用诱饵诱集法研究了荒地、绿化带生境中红火蚁蚁丘体积、蚁丘表面基部面积与诱集工蚁数量之间的关系.结果表明：在荒地生境中,随着蚁丘体积的增大,诱集工蚁数量表现为先迅速增加、后增大减缓、最终趋于平衡的规律,当蚁丘体积、蚁丘表面基部面积分别为11634 cm³、1308 cm²时,诱集工蚁数达到最大值,为每诱饵291 头;在绿化带生境中,随着蚁丘体积的增大,诱集工蚁数量动态与荒地生境相似,当蚁丘体积>18089 cm³时,诱集工蚁数量的增幅明显变缓,当蚁丘体积为25974 cm³时,诱集工蚁数达到最大值,为每诱饵232头.韦布尔函数方程可较好地描述诱集的红火蚁工蚁数量与蚁丘体积、蚁丘表面基部面积间的关系.     

人重组IL6／IL2融合蛋白的变性、复性及纯化

经超声破碎,分离已表达CH925包涵体,较系统地研究变性剂浓度、融合蛋白浓度对蛋白折叠的影响.在还原型及氧化型谷胱甘肽复性条件下,成功地将融合蛋白CH925折叠成具有IL6及IL2双活性蛋白,IL6的比活为2.3×10⁸U/mg, IL2比活为2.2×10⁶U/mg.经阴离子交换、凝胶过滤层析,获得一定纯度的CH925,配合反相HPLC.洗脱收集蛋白峰,CH925纯度为98%.