CYP17A1基因新突变致17α-羟化酶/17,20-裂解酶部分性缺陷症

目的研究1例17α-羟化酶/17,20-裂解酶部分性联合缺陷症患者CYP17A1基因突变特点,并结合患者的临床表现与基因突变类型初步探讨P450C17酶蛋白的结构与功能的关系。方法收集1例17α-羟化酶/17,20-裂解酶部分性联合缺陷症患者的临床资料及其亲属血标本,提取基因组DNA,设计7对引物扩增CYP17A1基因的8个外显子及外显子与内含子的连接区域,琼脂糖凝胶电泳鉴定PCR产物,产物胶回收后直接做为DNA双链模板测序。DNA双链模板不一致的PCR产物经克隆后测序。测序结果在核苷酸序列数据库进行比较分析。结果患者CYP17A1基因突变检测结果为5994-5995delAT/7541C>T复合杂合子。这两种突变均未见报道。推测5994-5995delAT导致I259H,274X,突变形成的截短蛋白质缺少血红素结合区域,因此是没有功能的;而通过人类P450C17酶计算机模型分析显示7541C>T导致的A398V远离酶的活性中心,推测突变可能使酶的活性减弱,而不是完全地丧失。患者临床表现为有自发不规则月经及轻度高血压、低血钾,结合激素测定结果提示肾上腺和性腺保留部分功能。因而患者的基因型与其临床表型是一致的。结论应进行突变P450C17酶的功能学研究来进一步明确结构改变对功能的影响。     

Evaluation of cleaving agents other than trypsin in direct agglutination test for further improving diagnosis of visceral leishmaniasis.

Trypsin treatment of Leishmania promastigote antigen has proved to be indispensible in the direct agglutination test (DAT) for the diagnosis of visceral leishmaniasis (VL) and canine visceral leishmaniasis (CVL). In the present study four antigen batches were prepared with pronase (400 micrograms/ml), lipase (0.45% [wt/vol]), pancreatin (0.3% [wt/vol]), or 2-mercaptoethanol (2-ME) (1.2% [vol/vol]) at a ratio of 20:1 versus promastigote packed cell volume or a density of 10(8)/ml. Batches prepared in this way performed satisfactorily when compared with the performance of the initial trypsinated antigen. Even higher was the sensitivity and specificity of the 2-ME-processed antigen, scoring a minimum DAT titer of 1:102,400 in the VL and CVL group and a maximum of 1:400 in the negative control group. Corresponding titers ranging from 1:6,400 to 1:12,800 and 1:800 to 1:1,600 were obtained with the antigen variants processed with pronase, lipase, pancreatin, or trypsin. By combining the use of indigenous Leishmania donovani subspecies from Sudan, Bangladesh, or Morocco and incorporating 2-ME instead of trypsin in the antigen processing step, a threefold increase in titer was attained in sera from the respective areas where VL is endemic. 2-ME-processed antigen suspensions maintained stability at 4 degrees C for up to 9 months, as evidenced by the absence of autoagglutination and the reproducibility of DAT readings with standard sera. The specificity of DAT was further improved by supplementation of the sample diluent with 0.03 M urea and incubation of the test plates at 37 degrees C for 1 h.(ABSTRACT TRUNCATED AT 250 WORDS)     

A-kinase anchoring proteins in amygdala are involved in auditory fear memory

A-kinase anchoring proteins (AKAPs) constitute a family of scaffolding proteins that bind the regulatory subunits of protein kinase A (PKA). AKAP binding to PKA regulates the phosphorylation of various proteins, some of which have been implicated in synaptic plasticity and memory consolidation. Here we show that the regulatory subunits of PKA are colocalized with AKAP150 (an AKAP isoform that is expressed in the brain) in the lateral amygdala (LA) and that infusion to the LA of the peptide St-Ht31, which blocks PKA anchoring onto AKAPs, impairs memory consolidation of auditory fear conditioning.     

Longitudinal growth of Bangladeshi infants during the first year of life

The growth in length and weight of 91 poor urban Bangladeshi infants was monitored at monthly intervals from birth to 1 year. At birth 18%, 22% and 8% were below -2.00 standard deviations of the National Center for Health Statistics (NCHS) height-for-age, weight-for-age and weight-for-height, respectively and at 1 year the percentages were 40%, 40% and 7%. Infant weights over the first 6 months associated positively with both mother's BMI and percentage body fat. Higher mean infant weights from 6 to 12 months were associated with both higher educational status of the mother and monthly family income and the latter two variables provided the best model for predicting weight velocity from birth to 12 months. Internal Z-scores, corrected for day of measurement, provided clear evidence of catch-up/catch-down over the first 6 months with heavier and longer babies at birth showing catch-down while lighter and shorter babies demonstrated catch-up. Infants' weights were almost three times more variable than lengths. Monthly incremental variability of both infant weight and length increased sharply from 6 to 12 months with weight showing significantly more variability. The correlations of birth weight and length with subsequent distances and monthly increment revealed that the first 6 months were dominated by catch-up and catch-down but during the latter half of the year the growth phenomena were influenced mainly by earlier intra-uterine or genetic effects.     

Persistence of hepatitis B antigen in Culex quinquefasciatus (Diptera: Culicidae)

Groups of wild-caught Culex quinquefaciatus Say, previously tested for the presence of hepatitis B surface antigen (HBsAg), were tested for the presence of hepatitis B e antigen (HBeAg). This antigen was detected at low levels in blood-fed, half-gravid, and gravid groups. A colony of Cx. quinquefasciatus was established in the laboratory and tested for the persistence of HBsAg and HBeAg. Five days after feeding on blood infected with HBsAg and HBeAg, 9 of 20 (45%) pools of Cx. quinquefasciatus were HBeAg-positive and 5 of 20 (25%) pools were HBeAg-positive; low levels of HBsAg and HBeAg were still detectable 28 d after the infective meal in 2 of 20 (10%) and 1 of 20 (5%) pools, respectively. A crude protease extract was prepared from colony mosquitoes, and the effect of this extract on HBsAg and HBeAg present in human serum was tested in vitro. After 20 h, tests for both antigens were still strongly positive. Low levels of HBsAg were detected in ovaries 7 d after infection. Salivary glands were HBsAg- and HBeAg-negative.     

Feasibility study on the utilization of rubber latex effluent for producing bacterial biopolymers

Rubber latex effluent is a polluting source that has a high biochemical oxygen demand (BOD). It is estimated that about 100 million liters of effluent are discharged daily from rubber processing factories. Utilization of this effluent such as the use of a coupled system not only can reduce the cost of treatment but also yield a fermentation feedstock for the production of bioplastic. This study initially was carried out to increase the production of organic acids by anaerobic treatment of rubber latex effluent. It was found that through anaerobic treatment the concentration of organic acids did not increase. Consequently, separation of organic acids from rubber latex effluent by anion exchange resin was examined as a preliminary study of recovering acetic and propionic acids. However, the suspended solids (SS) content in the raw effluent was rather high which partially blocked the ion-exchange columns. Lime was used to remove the SS in the rubber latex effluent. After the lime precipitation process, organic acids were found to adsorb strongly onto the anion exchange resin. Less adsorption of organic acids onto the resin was observed before the lime precipitation. This was probably due to more sites being occupied by colloidal particles on the resin thus inhibiting the adsorption of organic acids. The initial concentration of organic acids in the raw effluent was 3.9 g/L. After ion exchange, the concentration of the organic acids increased to 27 g/L, which could be utilized for production of polyhydroxyalkanoates (PHA). For PHA accumulation stage, concentrated rubber latex effluent obtained from ion exchange resins and synthetic acetic acid were used as the carbon source. Quantitative analyses from fed batch culture via HPLC showed that the accumulation of PHA in Alcaligenes eutrophus was maximum with a concentration of 1.182 g/L when cultivated on synthetic acetic acid, corresponding to a yield of 87% based on its cell dry weight. The dry cell weight increased from 0.71 to 1.67 g/L. On the other hand, using concentrated rubber latex effluent containing acetic and propionic acids resulted in reduced PHA content by dry weight (14%) but the dry cell weight increased from 0.49 to 1.30 g/L. The results clearly indicated that the cells grow well in rubber latex effluent but no PHA was accumulated. This could be due to the high concentration of propionic acid in culture broth or other factors such as heavy metals. Thus further work is required before rubber latex effluent can be utilized as a substrate for PHA production industrially.     

Supra-maximal cycling efficiency assessed in humans by using a new protocol

This study proposed a non-invasive method to determine the gross (GE, no baseline correction), net (NE, resting metabolism as the baseline correction) and work (WE, unloaded cycling as the baseline correction) efficiencies during cycling at an intensity higher than the maximal aerobic power (MAP). Twelve male subjects performed two exercises consisting of 4 min at 50% MAP followed either by 8 min at 63% MAP or by 8 sequences of 60 s divided into 10 s at 130% MAP and 50 s at 50% MAP (i.e., 63% MAP on average). Oxygen uptake was continuously measured to calculate GE, NE and WE at 50%, 63% and 130% MAP, and the data presented as the means and standard deviations. The GE values were 18.2%, 19.1%, 22.7%, the NE values were 22.4%, 22.8%, 24.3% and the WE values were 34.2%, 31.4% and 27.2% at 50%, 63% and 130% MAP, respectively. The GE and NE increased (P<0.001) whereas the WE decreased (P<0.001) with each increment in power output. The GE was lower than the NE (P<0.001) at 50% and 63% MAP and than the WE (P<0.001) at all intensities. The NE was lower (P<0.001) than the WE at 50% and 63% MAP. These results showed that (1) efficiency index values obtained during supra-maximal exercise were consistent with previous proposals and (2) the efficiency-power output relationships were not limited to sub-maximal intensity levels but were confirmed at higher power output.     

Identification and characterization of human pulmonary dendritic cells

Dendritic cells (DC) are specialized antigen-presenting cells, linking innate and adaptive immune responses, and thus play an important role in immunologically mediated diseases, including pulmonary diseases such as asthma and respiratory viral infections. Although much is known about the characteristics of lung DC in animal models, very few data concerning human lung DC are available. The goal of our study was to identify and characterize dendritic cells in human lung by preparing single-cell suspensions from surgical resection specimens and subsequent labeling with the recently developed blood dendritic cell antigen (BDCA) markers. A straightforward isolation procedure was developed to avoid phenotypical and functional changes induced by extensive purification methods. In this way, human lung DC were directly identified without the need for an additional adherence step for further purification. For the first time, we demonstrate the presence of three previously unidentified DC subsets in human lung digests: myeloid DC type 1 (BDCA1+/HLA-DR+), myeloid DC type 2 (BDCA3+/HLA-DR+), and plasmacytoid DC (BDCA2+/CD123+). The presence of CD1a+ DC in the human lung was confirmed. The identification and characterization of different human pulmonary DC subtypes is of great importance for the future development of DC-based immunotherapies.     

Wasting during pregnancy increases the risk of mother-to-child HIV-1 transmission

OBJECTIVES: To examine whether wasting during pregnancy, as measured by weight loss and low weight gain, is associated with increased mother-to-child transmission (MTCT) of HIV-1. METHODS: This was a cohort study in Dar es Salaam, Tanzania, among 957 HIV-1-infected pregnant women. Weight was measured at the first prenatal visit and every month thereafter until delivery. Weight loss was defined as a weekly rate of weight gain 0 and /=167 g/wk, weight loss during pregnancy was related to higher risk of intrauterine MTCT (adjusted relative risk [RR] = 2.32, 95% CI = 1.23-4.36, P = 0.009), HIV positive at birth or fetal death (RR = 2.13, 95% CI = 1.40-3.24, P = 0.0004), and HIV positive at birth or early neonatal death (RR = 1.96, 95% CI = 1.26-3.07, P = 0.003). The rate of weight gain during the 3rd trimester was inversely related to the risk of intrapartum/early breast-feeding transmission (adjusted P value, test for trend = 0.05). CONCLUSIONS: Weight loss during pregnancy increases the risk of early MTCT. Identifying causes of wasting during pregnancy may provide clues for new strategies to prevent MTCT.