Metabolism of nitric oxide by Neisseria meningitidis modifies release of NO-regulated cytokines and chemokines by human macrophages

Macrophages produce nitric oxide (NO) via the inducible nitric oxide synthase as part of a successful response to infection. The gene norB of Neisseria meningitidis encodes a NO reductase which enables utilization and consumption of NO during microaerobic respiration and confers resistance to nitrosative stress-related killing by human monocyte-derived macrophages (MDM). In this study we confirmed that NO regulates cytokine and chemokine release by resting MDM: accumulation of TNF-alpha, IL-12, IL-10, CCL5 (RANTES) and CXCL8 (IL-8) in MDM supernatants was significantly modified by the NO-donor S-nitroso-N-penicillamine (SNAP). Using a protein array, infection of MDM with N. meningitidis was shown to be associated with secretion of a wide range of cytokines and chemokines. To test whether NO metabolism by N. meningitidis modifies release of NO-regulated cytokines, we infected MDM with wild-type organisms and an isogenic norB strain. Resulting expression of the cytokines TNF-alpha and IL-12, and the chemokine CXCL8 was increased and production of the cytokine IL-10 and the chemokine CCL5 was decreased in norB-infected MDM, in comparison to wild-type. Addition of SNAP to cultures infected with wild-type mimicked the effect observed in cultures infected with the norB mutant. In conclusion, NorB-catalysed removal of NO modifies cellular release of NO-regulated cytokines and chemokines.     

神经内科硕士研究生临床和科研能力的培养

临床医学硕士研究生阶段是培养临床和科研思维能力的重要阶段,从某种意义上说是人生和医疗生涯的关键时期。神经内科硕士研究生不仅应该具有一定的临床工作能力,还应具有一定的科研能力。临床工作能力培养包括临床基本功的培训,基本理论的加强和基本技能的培养,临床思维能力的培养及医患沟通能力的培养等多个方面。科研能力包括文献阅读能力、科研思维能力和论文写作能力等。这样,硕士研究生毕业后不仅能够诊治神经内科常见病和多发病,会思考临床工作中的问题,更重要的是能想办法探索和解决这些问题。这也是硕士研究生和本科生的本质区别。因此,研究生阶段是医学生涯一个重要的里程碑,临床和科研能力的培养对个人未来的发展具有十分重要的意义。     

Virulence in malaria: an evolutionary viewpoint

Malaria parasites cause much morbidity and mortality to their human hosts. From our evolutionary perspective, this is because virulence is positively associated with parasite transmission rate. Natural selection therefore drives virulence upwards, but only to the point where the cost to transmission caused by host death begins to outweigh the transmission benefits. In this review, we summarize data from the laboratory rodent malaria model, Plasmodium chabaudi, and field data on the human malaria parasite, P. falciparum, in relation to this virulence trade-off hypothesis. The data from both species show strong positive correlations between asexual multiplication, transmission rate, infection length, morbidity and mortality, and therefore support the underlying assumptions of the hypothesis. Moreover, the P. falciparum data show that expected total lifetime transmission of the parasite is maximized in young children in whom the fitness cost of host mortality balances the fitness benefits of higher transmission rates and slower clearance rates, thus exhibiting the hypothesized virulence trade-off. This evolutionary explanation of virulence appears to accord well with the clinical and molecular explanations of pathogenesis that involve cytoadherence, red cell invasion and immune evasion, although direct evidence of the fitness advantages of these mechanisms is scarce. One implication of this evolutionary view of virulence is that parasite populations are expected to evolve new levels of virulence in response to medical interventions such as vaccines and drugs.     

Assessment and management of old-growth forests in south eastern Australia

Abstract

Old-growth forests in south eastern Australia are important for biodiversity conservation, recreation, carbon storage, social values and, to a declining extent, for timber production. Developing a comprehensive definition of old-growth forest that can apply across all Australian vegetation types has been challenging. Old growth can be viewed from ecological and social perspectives. For policy and management purposes old growth has been defined as a growth stage in forest development, incorporating ecological maturity and lack of evidence of past disturbance. Classification and assessment of old growth has largely been restricted to those areas covered by regional forest agreements (RFAs) between different states and the Federal Government. Old growth can be impacted by wildfire, timber harvesting, insect pests, diseases and other disturbances. Climate change will also present challenges for the future management of old-growth forests. There is increasing scientific understanding of the relationships between species, forest growth stage and old-growth forest attributes. To meet biodiversity conservation objectives, the management focus is shifting from assessing and protecting old-growth forests, to providing for forests across the landscape with old-growth attributes. This approach may be at odds with other conceptions of old growth based on notions of undisturbed systems free of human influence.     

基于钙黄绿素的荧光分光光度法测定谷胱甘肽

目的:基于钙黄绿素的荧光分光光度法建立一种测定谷胱甘肽的新方法。方法:在pH=8.0介质中,铜(Ⅱ)与钙黄绿素配位引起荧光猝灭,由于谷胱甘肽与铜(Ⅱ)的亲和力很强,可从钙黄绿素-铜离子的络合物中夺取铜离子而使钙黄绿素游离出来使荧光强度得以恢复,荧光恢复的程度与加入谷胱甘肽的浓度在一定浓度范围内成线性。结果:据此建立了一种测定谷胱甘肽的新方法,该方法的线性范围为2.0×10-6～1.4×10-5mol.L-1,F=8.1964C+196.43,检测限为1.0×10-6mol/L。结论:用此法测定谷胱甘肽简便快速,灵敏度高。     

Identification of MHC class I sequences in Chinese-origin rhesus macaques

The rhesus macaque (Macaca mulatta) is an excellent model for human disease and vaccine research. Two populations exhibiting distinctive morphological and physiological characteristics, Indian- and Chinese-origin rhesus macaques, are commonly used in research. Genetic analysis has focused on the Indian macaque population, but the accessibility of these animals for research is limited. Due to their greater availability, Chinese rhesus macaques are now being used more frequently, particularly in vaccine and biodefense studies, although relatively little is known about their immunogenetics. In this study, we discovered major histocompatibility complex (MHC) class I cDNAs in 12 Chinese rhesus macaques and detected 41 distinct Mamu-A and Mamu-B sequences. Twenty-seven of these class I cDNAs were novel, while six and eight of these sequences were previously reported in Chinese and Indian rhesus macaques, respectively. We then performed microsatellite analysis on DNA from these 12 animals, as well as an additional 18 animals, and developed sequence specific primer PCR (PCR-SSP) assays for eight cDNAs found in multiple animals. We also examined our cohort for potential admixture of Chinese and Indian origin animals using a recently developed panel of single nucleotide polymorphisms (SNPs). The discovery of 27 novel MHC class I sequences in this analysis underscores the genetic diversity of Chinese rhesus macaques and contributes reagents that will be valuable for studying cellular immunology in this population.     

功能矫形前伸青春期大鼠下颌后浅层嚼肌细胞凋亡的研究

目的:研究功能矫形前伸大鼠下颌后浅层嚼肌细胞凋亡的变化规律,探讨功能矫形的肌肉改建机理。方法:选用50只5周龄Sprague-Dawley(SD)雄性大白鼠,随机分为实验组和对照组各25只。实验组大鼠戴自制上颌功能矫治嚣,引导下颌前伸,并打开咬合。利用RT-PCR方法检测两组大鼠浅层嚼肌Bcl-2和Bax基因表达情况,利用TUNEL方法检测浅层嚼肌细胞凋亡情况。结果:①Bcl-2和Bax基因表达随大鼠戴用矫治器时间的延长而升高,至第3周开始下降但仍高于对照组,但Bax的表达高于Bcl-2。Bax/Bcl-2比值随大鼠戴用矫治器时间的延长而升高,至第4周开始下降。②TUNEL实验结果显示浅层嚼肌细胞在戴用矫治器1天后,开始出现凋亡,随着时间延长而增加,至第3周达到顶峰,第4周开始下降。结论:①Bax/Bcl-2比值升高促进浅层嚼肌细胞凋亡。②功能矫形可引起浅层嚼肌细胞凋亡,导致肌肉的结构和功能发生适应性改建。     

米槠-木荷林优势种群的年龄结构及其更新策略

通过年轮分析,构建了浙江天童米槠-木荷林优势种木荷、米槠、石栎的年龄结构,分析了高生长和径向生长及萌枝策略随年龄的变化规律,探讨了干扰对群落动态的影响。研究结果表明:(1)3个物种的胸径、高度与年龄的关系均可用logistic曲线和线性模型拟合,但随年龄增加胸径、高度与年龄的关系显著下降。(2)随年龄增加3树种的DBH/H逐渐增加,米槠、石栎幼龄个体(1-20a)的有萌个体率和有萌个体萌枝数逐渐下降;米槠和石栎幼苗的主干较木荷细长、多萌枝,木荷和石栎成树主干较米槠细长;体现了米槠的更新策略具开拓性,木荷的具保守性,石栎在幼龄期具开拓性,成年期转为保守性。(3)米槠和石栎的年龄结构呈逆J型,幼龄个体充足,但中龄段(21-40a)存在更新断层,这与其高消耗的开拓性策略有关;木荷的年龄结构呈间歇型,各龄级均存有个体,这与其保守性策略有关;(4)3物种在大龄级上(52-60a)均存在更新高峰,与该地区的择伐干扰时间一致,高强度的干扰促生了次生演替,形成了以木荷为第一优势种的群落。(5)米槠、木荷、石栎的年龄结构是更新策略和干扰的综合表现,它不仅与径级结构一样可展现种群更新特征、预测种群发展动态,更能精确地反映群落动态事件的发生频次和发生时间。     

Studies on the mechanism for entry of vesicular stomatitis virus glycoprotein g mRNA into membrane-bound polyribosome complexes

Glycoprotein mRNA (G mRNA) of vesicular stomatitis virus is synthesized in the cytosol fraction of infected HeLa cells. Shortly after synthesis, this mRNA associates with 40S ribosomal subunits and subsequently forms 80S monosomes in the cytosol fraction. The bulk of labeled G mRNA is then found in polysomes associated with the membrane, without first appearing in the subunit or monomer pool of the membrane-bound fraction. Inhibition of the initiation of protein synthesis by pactamycin or muconomycin A blocks entry of newly synthesized G m RNA into membrane-bound polysomes. Under these circumstances, labeled G mRNA accumulates into the cytosol. Inhibition of the elongation of protein synthesis by cucloheximide, however, allows entry of 60 percent of newly synthesized G mRNA into membrane-bound polysomes. Furthermore, prelabeled G mRNA associated with membrane-bound polysomes is released from the membrane fraction in vivo by pactamycin or mucomycon A and in vitro by 1mM puromycin - 0.5 M KCI. This release is not due to nonspecific effects of the drugs. These results demonstrate that association of G mRNA with membrane-bound polysomes is dependent upon polysome formation and initiation of protein synthesis. Therefore, direct association of the 3' end of G mRNA with the membrane does not appear to be the initial event in the formation of membrane-bound polysomes.     

Computer assisted self interviewing in a sexual health clinic as part of routine clinical care; impact on service and patient and clinician views

Background

Computer assisted self interviewing (CASI) has been used at the Melbourne Sexual Health Centre (MSHC) since 2008 for obtaining sexual history and identifying patients'' risk factors for sexually transmitted infections (STIs). We aimed to evaluate the impact of CASI operating at MSHC.

Methodology/Principal Findings

The proportion of patients who decline to answer questions using CASI was determined. We then compared consultation times and STI-testing rates during comparable CASI and non-CASI operating periods. Patients and staff completed anonymous questionnaires about their experience with CASI. 14,190 patients completed CASI during the audit period. Men were more likely than women to decline questions about the number of partners they had of the opposite sex (4.4% v 3.6%, p = 0.05) and same sex (8.9% v 0%, p<0.001). One third (34%) of HIV-positive men declined the number of partners they had and 11–17% declined questions about condom use. Women were more likely than men to decline to answer questions about condom use (2.9% v 2.3%, p = 0.05). There was no difference in the mean consultation times during CASI and non-CASI operating periods (p≥0.17). Only the proportion of women tested for chlamydia differed between the CASI and non-CASI period (84% v 88% respectively, p<0.01). 267 patients completed the survey about CASI. Most (72% men and 69% women) were comfortable using the computer and reported that all their answers were accurate (76% men and 71% women). Half preferred CASI but 18% would have preferred a clinician to have asked the questions. 39 clinicians completed the staff survey. Clinicians felt that for some STI risk factors (range 11%–44%), face-to-face questioning was more accurate than CASI. Only 5% were unsatisfied with CASI.

Conclusions

We have demonstrated that CASI is acceptable to both patients and clinicians in a sexual health setting and does not adversely affect various measures of clinical output.