Evaluation of Brugia malayi sheath protein (Shp-1) as a diagnostic antigen for human lymphatic filariasis

Lymphatic filariasis is the second leading cause of permanent long-term disability globally and control of this disease needs efficient diagnostic methods. In this study, abundantly expressing microfilarial sheath protein (Shp-1) from Brugia malayi was characterized as a filarial diagnostic candidate using samples from different clinical population. Monoclonal antibodies were developed against E. coil expressed recombinant Shp-1 in order to assess its efficiency in filarial antigen detection assay system. Endemic Normal (EN, n = 170), asymptomatic microfilaeremics (MF, n = 65), symptomatic chronic pathology (CP, n = 45) and non endemic normal (NEN, n = 10) sera were analyzed by antigen capture enzyme-linked immunosorbent assay. Of the 290 individuals, all MF individuals (both brugian and bancroftian) were positive in this assay followed by CP and EN. When compared with SXP-1 and Og4C3 antigen assays, all assays detected Wb MF correctly, Bm MF was detected by Shp-1 and SXP-1 assays, and only Shp-1 was able to detect EN (12%) and CP (29%). Results showed that this assay may be useful for monitoring prior to mass drug administration. (c) 2014 Elsevier Inc. All rights reserved.

Calcium binding properties of calcium dependent protein kinase 1 (CaCDPK1) from Cicer arietinum

Calcium plays a crucial role as a secondary messenger in all aspects of plant growth, development and survival. Calcium dependent protein kinases (CDPKs) are the major calcium decoders, which couple the changes in calcium level to an appropriate physiological response. The mechanism by which calcium regulates CDPK protein is not well understood. In this study, we investigated the interactions of Ca2+ ions with the CDPK1 isoform of Cicer arietinum (CaCDPK1) using a combination of biophysical tools. CaCDPK1 has four different EF hands as predicted by protein sequence analysis. The fluorescence emission spectrum of CaCDPK1 showed quenching with a 5 nm red shift upon addition of calcium, indicating conformational changes in the tertiary structure. The plot of changes in intensity against calcium concentrations showed a biphasic curve with binding constants of 1.29 mu M and 120 mu M indicating two kinds of binding sites. Isothermal calorimetric (ITC) titration with CaCl2 also showed a biphasic curve with two binding constants of 0.027 mu M and 1.7 mu M. Circular dichroism (CD) spectra showed two prominent peaks at 208 and 222 nm indicating that CaCDPK1 is a alpha-helical rich protein. Calcium binding further increased the alpha-helical content of CaCDPK1 from 75 to 81%. Addition of calcium to CaCDPK1 also increased fluorescence of 8-anilinonaphthalene-1-sulfonic acid (ANS) indicating exposure of hydrophobic surfaces. Thus, on the whole this study provides evidence for calcium induced conformational changes, exposure of hydrophobic surfaces and heterogeneity of EF hands in CaCDPK1. (C) 2015 Elsevier GmbH. All rights reserved.

Hypothetical protein Rv3423.1 of Mycobacterium tuberculosis is a histone acetyltransferase

We isolated an 8 kDa mycobacterial hypothetical protein, Rv3423.1, from the chromatin of human macrophages infected with Mycobacterium tuberculosis H37Rv. Bioinformatics predictions followed by in vitro biochemical assays with purified recombinant protein showed that Rv3423.1 is a novel histone acetyltransferase that acetylates histone H3 at the K9/K14 positions. Transient transfection of macrophages containing GFP-tagged histone H1 with RFP-tagged Rv3423.1 revealed that the protein co-localizes with the chromatin in the nucleus. Co-immunoprecipitation assays confirmed that the Rv3423.1-histone interaction is specific. Rv3423.1 protein was detected in the culture filtrate of virulent but not avirulent M. tuberculosis. Infection of macrophages with recombinant Mycobacterium smegmatis constitutively expressing Rv3423.1 resulted in a significant increase in the number of intracellular bacteria. However, the protein did not seem to offer any growth advantage to free-living recombinant M. smegmatis. It is highly likely that, by binding to the host chromatin, this histone acetyltransferase from M. tuberculosis may manipulate the expression of host genes involved in anti-inflammatory responses to evade clearance and to survive in the intracellular environment.

Individual weight, development time and yield of different stages of Tropodiaptomus incognitus (Crustacea: Copepoda). [Translation from: Cahiers ORSTOM, Serie Hydrobiologie 4(1) 63-70, 1970. ]

Observations of individual weight, duration of development and production of different stages of Tropodiaptomus incognitus are presented. The study is based on data gathered from Lake Chad in 1968.

含反-1,4聚丁二烯链段的顺-1,4聚丁二烯橡胶的合成、（金+监）认和性能

顺-1,4聚丁二烯橡胶是合成橡胶中的第二大品种，具有耐磨、生热低等的优点，一贯是轮胎胎面胶的重要组成部分。但随着子午线轮胎的出现和大量使用，对母胶强度和轮胎的耐湿滑性提出了更高的要求，而利用应力下结晶有可能提高母胶强度，为此我们开展了合成含少量反-1，4聚丁二烯链烯链段的顺-1，4-聚丁二烯橡胶的研究。本文报导了它的合成、（金+监）认及生胶、母胶和硫化胶的性能。合成中我们试探了既能使丁二烯反-1，4聚合和顺-1，4聚合，又能使内双键打开的钒和钴的络合催化体系，采用以钒催化剂先使丁二烯反-1，4聚合，在达到转化率10-20%时，以钴催化剂将剩余的丁二烯进行顺-1，4聚合的步骤，再寄于同时发生催化接枝的想法来实现反-1，4-聚丁二烯链段和顺-1，4聚丁二烯链段的共聚合。实验中考查了合成的条件，对得到的产物用溶解度法、扭摆测定动态力学行为，X射线衍射分析、电镜分析以及线膨胀等手段和以等量的反-1，4聚丁二烯和顺-1，4聚丁二烯的共混胶进行了对比，证明我们得到的产物是反-1，4聚丁二烯链段和顺-1，4聚丁二烯的接枝共聚物，不是无规共聚物，也不是两种均聚物的共混胶。文中讨论了聚合反应机理。对含有不同反-1，4链段和分子量的共聚胶的生胶性能，混炼胶性能进行了初步考察，结果表明，特性粘数（[η]）_(甲苯）~（30℃）在2（分升/克）左右，含反-1，4链段9-20%的顺丁共聚胶，不含凝胶，其生胶强度和混炼胶强度较镍顺丁橡胶高一倍以上，挤出行为特好。看来这种共聚胶将为子午线轮胎所欢迎的新顺丁品种。对这种胶的硫化配方和硫化条件条件尚待进一步研究。初步结果表明，可与一般顺丁橡胶接近，而抗撕性能又比一般顺丁高。利用本工作的实验方法制备不同结构链段的共聚胶还不多见。因此，除了工作本身研制成了一种新的顺丁品种外，对利用本方法，改善其他橡胶品种，设计研制新的高分子和研究反应机理走出了一条新路。

0.4-1.2A GeV能区重离子碰撞中的直接流系统性研究

直接流是研究重离子碰撞动力学演变和压缩形成的高密核物质性质很好的探针，本论文系统地研究了0.4、0.8和1.16A GeV的Ni+Ni和Pb+Pb碰撞中的直接流。实验是在德国重离子研究中心(GSI)的FOPI探测装置上完成的。论文中，简单总结了中高能区重离子碰撞的现状和描述集体流的主要理论模型，介绍了FOPI探测系统，给出了详细的实验数据分析过程，对所得到的物理结果进行了讨论。本论文工作的重点如下：基于FOPI系统的实验数据，发展了一套质量相关(Z=1离子)的直接流的提取方法。提取了各碰撞系统出射P、D和T粒子在不同碰撞中心度下的微分直接流和积分直接流。研究了P、D和T的直接流对碰撞中心度、系统尺寸和碰撞能量的依赖性，以及对核物质状态方程的敏感性。结果表明：直接流敏感地依赖于碰撞中心度，近中心碰撞具有更强直接流信号；对于轻重两种系统，用常用的AP1/3+AT1/3系数对积分直接流进行了标度，观察到一定的标度性，但不能完全标度；通过研究直接流对碰撞能量的依赖性发现，在0.4－1.2A GeV能区内，随能量升高，直接流在已经达到了饱和，并开始下降，并且P、D和T的变化趋势相同。实验数据与输运模型IQMD计算比较发现，直接流的变化趋势和最大密度变化趋势相同，说明直接流是核物质压缩程度的一个良好探针。计算得到的P、D和T微分和积分的V1值表明，与质量相关的直接流，无论是微分值还是积分值都敏感依赖于模型中EoS参数。比较发现，不同碰撞能量下，重的Pb+Pb系统的数据和软的EoS符合很好，说明核物质不可压缩系数在210 MeV附近，这与文献中的结果相吻合，说明与质量相关的直接流是EoS的敏感探针。对于轻Ni＋Ni系统，目前的IQMD还不能重现数据，但其趋向于硬的EoS，需要发展描述碰撞过程更为精细的理论模型。数据整体趋势表明，随者系统变重，中子比例的增加，EoS变软，难以给出同一组IQMD参数来同时解释全部的实验数据。对于所研究的碰撞系统，比较中心快度区斜率行为时发现，P、D和T的直接流与出射粒子质量数呈线性关系，并且出射粒子的积分直接流可以很好的用常数(A+1)/2进行标度。如果出射粒子的直接流用IQMD计算的核阻止进行归一，归一后的直接流与碰撞能量成正比。这证明核阻止与直接流有线性关联，反映了核阻止对于碰撞中核物质达到的最高密度起决定性的作用。论文工作的另一部分是完成了FOPI探测装置中飞行时间探测器的升级工作。研制了新型的玻璃MMRPC，完成了性能的批量测试，并研究了该探测器的高计数率行为。测量结果显示，在实验计数率(0.1 kHz/cm2)条件下，MMRPC时间分辨达到75 ps，探测效率达到98％。当计数率达到3－5 kHz/cm2时，时间分辨和探测效率降至约110 ps和75％。高计数率探测效率变差的幅度可以用DC模型进行解释，然而时间分辨的变化幅度用DC模型难以解释

联萘衍生物的NMR表征Ⅱ.2,2′-(3,4-四酸二酐)二苯甲酰氧基-1,1′联萘

用一维 1HNMR、13CNMR方法研究了2,2′-(3,4 -四酸二酐)二苯甲酰氧基 -1,1′联萘的结构 ,并通过二维1H - 1H同核相关、13C - 1H异核相关及13C - 1H异核远程相关谱进一步地确定其1H谱和13C谱中各谱峰的归属 ,为同类化合物的表征提供了依据。

稀土3,4-呋喃二甲酸,1,10-二氮杂菲配合物的光物理性质

研究了稀土Ｇｄ３＋、Ｅｕ３＋、Ｔｂ３＋的３，４呋喃二甲酸，１，１０二氮杂菲（Ｐｈｅｎ）配合物的合成、红外光谱、紫外光谱及光物理性质。详细讨论了配合物的分子内能量传递过程。发现分子内能量传递效率依赖于稀土中心离子的共振发射能级与配体最低三重态能级之间的相对位置。

Investigating the invasive and cytoprotective properties of the heme binding protein HRG-1 in cancer cells

This thesis investigates the mechanisms by which HRG-1 contributes to the invasive and cytoprotective signalling pathways in cancer cells through its effects on VATPase activity and heme transport. Plasma membrane-localised V-ATPase activity correlates with enhanced metastatic potential in cancer cells, which is attributed to extrusion of protons into the extracellular space and activation of pH-sensitive, extracellular matrix degrading-proteases. We found that HRG-1 is co-expressed with the V-ATPase at the plasma membrane of certain aggressive cancer cell types. Modulation of HRG-1 expression altered both the localisation and activity of the VATPase. We also found that HRG-1 enhances trafficking of essential transporters such as the glucose transporter (GLUT-1) in cancer cells, and increases glucose uptake, which is required for cancer cell growth, metabolism and V-ATPase assembly. Heme is potentially cytotoxic, owing to its iron moiety, and therefore the trafficking of heme is tightly controlled in cells. We hypothesised that HRG-1 is required for the transport of heme to intracellular compartments. Importantly, we found that HRG-1 interacts with the heme oxygenases that are necessary for heme catabolism. HRG-1 is also required for trafficking of both heme-bound and nonheme-bound receptors and suppression of HRG-1 results in perturbed receptor trafficking to the lysosome. Suppression of HRG-1 in HeLa cells increases toxic heme accumulation, reactive oxygen species accumulation, and DNA damage resulting in caspasedependent cell death. Mutation of essential heme binding residues in HRG-1 results in decreased heme binding to HRG-1. Interestingly, cells expressing heme-binding HRG-1 mutants exhibit decreased internalisation of the transferrin receptor compared to cells expressing wildtype HRG-1. These findings suggest that HRG- 1/heme trafficking contributes to a hitherto unappreciated aspect of receptormediated endocytosis. Overall, the findings of this thesis show that HRG-1-mediated regulation of intracellular and extracellular pH through V-ATPase activity is essential for a functioning endocytic pathway. This is critical for cells to acquire nutrients such as folate, iron and glucose and to mediate signalling in response to growth factor activation. Thus, HRG-1 facilitates enhanced metabolic activity of cancer cells to enable tumour growth and metastasis.