Arginine controls heat-induced cluster-cluster aggregation of lysozyme at around the isoelectric point

The process of protein aggregation has attracted a great deal of research attention, as aggregates are first of all a nuisance to preparation of high quality protein and secondly used as novel materials. In the latter case, the process of protein aggregation needs to be controlled. Here, we show how arginine (Arg) regulates the process of heat-induced protein aggregation. Dynamic light scattering and transmission electron microscopy revealed that heat-induced aggregation of lysozyme at around the isoelectric point occurred in a two-step process: formation of start aggregates, followed by further growth mediated by their sticking with diffusion-limited cluster-cluster aggregation. In the presence of Arg, the diffusion-limited regime changed to reaction-limited cluster-cluster aggregation. The data indicated that the solution additives that coexisted with proteins would affect the property of the formed product, such as morphology and mechanic strength.     

Lipid droplets formation in human endothelial cells in response to polyunsaturated fatty acids and 1‐methyl‐nicotinamide (MNA); confocal Raman imaging and fluorescence microscopy studies

In this work the formation of lipid droplets (LDs) in human endothelial cells culture in response to the uptake of polyunsaturated fatty acids (PUFAs) was studied. Additionally, an effect of 1‐methylnicotinamide (MNA) on the process of LDs formation was investigated. LDs have been previously described structurally and to some degree biochemically, however neither the precise function of LDs nor the factors responsible for LD induction have been clarified. Lipid droplets, sometimes referred in the literature as lipid bodies are organelles known to regulate neutrophil, eosinophil, or tumor cell functions but their presence and function in the endothelium is largely unexplored. 3D linear Raman spectroscopy was used to study LDs formation in vitro in a single endothelial cell. The method provides information about distribution and size of LDs as well as their composition. The incubation of endothelial cells with various PUFAs resulted in formation of LDs. As a complementary method for LDs identification a fluorescence microscopy was applied. Fluorescence measurements confirmed the Raman results suggesting endothelial cells uptake of PUFAs and subsequent LDs formation in the cytoplasm of the endothelium. Furthermore, MNA seem to potentiate intracellular uptake of PUFAs to the endothelium that may bear physiological and pharmacological significance.

Confocal Raman imaging of HAoEC cell with LDs.     

独叶草叶片性状表型多样性研究

独叶草是特产于中国的一种珍稀濒危植物.为研究独叶草自然居群的叶片形态变异特征及影响因素,选取分布于陕西、四川、甘肃的9个自然居群,对独叶草叶片的叶面积、叶柄长、总齿数、末级叶脉数、盲脉数、网结脉数等指标进行统计分析.结果表明:(1)独叶草叶片各属性间存在显著相关性,个别性状表现出与地理位置的相关性.(2)主成分分析显示独叶草的末级叶脉数、叶面积、总齿数对变异有较大贡献,而网结脉贡献很小.(3)独叶草叶片性状居群间变异程度大于居群内,其中盲脉数量在各个居群间变异程度最大,而末级叶脉数为最稳定的性状.(4)将本研究中形态变异结果与前期的遗传分析结果对比分析认为独叶草形态变异主要来自生态环境,而非遗传组成.(5)独叶草居群的形态及遗传变异聚类结果都显示独叶草形态变异并不具有地理格局规律,可能由于地质历史变化打断了独叶草原本连续的分布区,遗传保守性使其仍然保持分布区断裂前的遗传特性,但其表型随着生境的变化表现出可塑性.     

Crystal structure of a family 4 uracil-DNA glycosylase from Thermus thermophilus HB8

Uracil-DNA glycosylase (UDG; EC 3.2.2.-) removes uracil from DNA to initiate DNA base excision repair. Since hydrolytic deamination of cytosine to uracil is one of the most frequent DNA-damaging events in all cells, UDG is an essential enzyme for maintaining the integrity of genomic information. For the first time, we report the crystal structure of a family 4 UDG from Thermus thermophilus HB8 (TthUDG) complexed with uracil, solved at 1.5 angstroms resolution. As opposed to UDG enzymes in its other families, TthUDG possesses a [4Fe-4S] cluster. This iron-sulfur cluster, which is distant from the active site, interacts with loop structures and has been suggested to be unessential to the activity but necessary for stabilizing the loop structures. In addition to the iron-sulfur cluster, salt-bridges and ion pairs on the molecular surface and the presence of proline on loops and turns is thought to contribute to the enzyme's thermostability. Despite very low levels of sequence identity with Escherichia coli and human UDGs (family 1) and E.coli G:T/U mismatch-specific DNA glycosylase (MUG) (family 2), the topology and order of secondary structures of TthUDG are similar to those of these distant relatives. Furthermore, the coordinates of the core structure formed by beta-strands are almost the same. Positive charge is distributed over the active-site groove, where TthUDG would bind DNA strands, as do UDG enzymes in other families. TthUDG recognizes uracil specifically in the same manner as does human UDG (family 1), rather than guanine in the complementary strand DNA, as does E.coli MUG (family 2). These results suggest that the mechanism by which family 4 UDGs remove uracils from DNA is similar to that of family 1 enzymes.     

Insights into the role of components of the tumor microenvironment in oral carcinoma call for new therapeutic approaches

The research on oral cancer has focused mainly on the cancer cells, their genetic changes and consequent phenotypic modifications. However, it is increasingly clear that the tumor microenvironment (TME) has been shown to be in a dynamic state of inter-relations with the cancer cells. The TME contains a variety of components including the non-cancerous cells (i.e., immune cells, resident fibroblasts and angiogenic vascular cells) and the ECM milieu [including fibers (mainly collagen and fibronectin) and soluble factors (i.e., enzymes, growth factors, cytokines and chemokines)]. Thus, it is currently assumed that TME is considered a part of the cancerous tissue and the functionality of its key components constitutes the setting on which the hallmarks of the cancer cells can evolve. Therefore, in terms of controlling a malignancy, one should control the growth, invasion and spread of the cancer cells through modifications in the TME components. This mini review focuses on the TME as a diagnostic approach and reports the recent insights into the role of different TME key components [such as carcinoma-associated fibroblasts (CAFs) and inflammation (CAI) cells, angiogenesis, stromal matrix molecules and proteases] in the molecular biology of oral carcinoma. Furthermore, the impact of TME components on clinical outcomes and the concomitant need for development of new therapeutic approaches will be discussed.     

应用SSR标记分析鹰嘴豆种质资源遗传多样性

种质资源的遗传多样性是育种工作的基础,本研究利用SSR标记对鹰嘴豆资源进行遗传多样性分析,旨在发掘鹰嘴豆资源中丰富的遗传变异。从48对鹰嘴豆SSR引物中筛选出18对核心多态性引物,对96份不同来源的鹰嘴豆种质资源进行SSR标记的遗传多样性分析。结果表明,18对SSR引物共检测到115个等位变异,占总检测位点的52.99%,每对SSR引物可检测出3~10个等位变异,平均6.39个;平均每个位点多态信息量(PIC)为0.8107,变化范围为0.6661~0.8984。Shannon's信息指数平均为1.4769,变化范围为0.0607~1.9584。PGMA聚类结果表明,在遗传相似系数0.59处,可将96份鹰嘴豆资源分为6个类群,类群Ⅰ包含9份资源,类群Ⅱ包含2份资源,类群Ⅲ包含28份资源,类群Ⅳ包含4份资源,类群Ⅴ包含40份资源,类群Ⅵ包含13份资源。本研究对我国鹰嘴豆种质资源的评价与利用、优异基因的挖掘、育种亲本材料的选择等提供科学依据。     

花生苗期耐盐性评价及耐盐指标筛选

评价鉴选耐盐品种对于盐碱地花生生产具有重要意义。采用盆栽试验,设置不同盐胁迫浓度,对200个花生品种(系)萌发至幼苗期通过出苗速度、植株形态和生物量等指标进行耐盐性系统评价。结果表明,随盐胁迫浓度的增加,出苗时间延长,植株形态建成抑制加重,物质积累减少。鉴选花生品种耐盐性强弱的适宜盐胁迫浓度为0.30%—0.45%,超过此浓度不能出苗。地上部形态和生物量可作为耐盐评价的首选指标,主根长和出苗速度可作为辅助指标用以判断花生品种的综合耐盐能力。200个品种(系)在不同盐胁迫浓度下均可分成高度耐盐型、耐盐型、盐敏感型和高度盐敏感型4组。耐盐品种数量随盐胁迫强度加大而下降,盐敏感品种数量则上升。0.15%浓度下200个品种全部出苗,4个类型品种数分别占供试材料的29.0%、39.0%、27.5%和4.5%;0.30%浓度下185个品种出苗,4个类型品种数分别占供试材料的5.5%、34.5%、23.5%和29.0%;0.45%浓度下107个品种出苗,4个类型品种数分别占供试材料的5.5%、5.5%、20.0%和22.5%。14个品种在各盐浓度胁迫下均表现耐性,10个品种在各盐浓度胁迫下均表现敏感,为花生耐盐机理研究及生产应用提供了不同类型材料。     

Cannabidiol as an emergent therapeutic strategy for lessening the impact of inflammation on oxidative stress

Oxidative stress with reactive oxygen species generation is a key weapon in the arsenal of the immune system for fighting invading pathogens and initiating tissue repair. If excessive or unresolved, however, immune-related oxidative stress can initiate further increasing levels of oxidative stress that cause organ damage and dysfunction. Targeting oxidative stress in various diseases therapeutically has proven more problematic than first anticipated given the complexities and perversity of both the underlying disease and the immune response. However, growing evidence suggests that the endocannabinoid system, which includes the CB₁ and CB₂ G-protein-coupled receptors and their endogenous lipid ligands, may be an area that is ripe for therapeutic exploitation. In this context, the related nonpsychotropic cannabinoid cannabidiol, which may interact with the endocannabinoid system but has actions that are distinct, offers promise as a prototype for anti-inflammatory drug development. This review discusses recent studies suggesting that cannabidiol may have utility in treating a number of human diseases and disorders now known to involve activation of the immune system and associated oxidative stress, as a contributor to their etiology and progression. These include rheumatoid arthritis, types 1 and 2 diabetes, atherosclerosis, Alzheimer disease, hypertension, the metabolic syndrome, ischemia-reperfusion injury, depression, and neuropathic pain.     

Comparing clustered sampling designs for spatially explicit estimation of population density

Spatially explicit capture–recapture methods do not assume that animals have equal access to sampling devices (e.g., detectors), which allows for gaps in the sampling extent and nonuniform (e.g., clustered) sampling designs. However, the performance (i.e., relative root mean squared error [RRMSE], confidence interval coverage, relative bias and relative standard error) of clustered detector arrays has not been thoroughly evaluated. I used simulations to evaluate the performance of various detector and cluster spacings, cluster configurations (i.e., number of detectors arranged in a square grid), sampling extents and number of sampling occasions for estimating population density, the relationship between detection rate and distance to a detector from the animal's center of activity (σ) and base detection rates, using American black bears (Ursus americanus) as a case study. My simulations indicated that a wide range of detector configurations can provide reliable estimates if spacing between detectors in clusters is ≥1σ and ≤3σ. A number of cluster configurations and occasion lengths produced estimates that were unbiased, resulted in good spatial coverage, and were relatively precise. Moreover, increasing the duration of sampling, establishing large study areas, increasing detection rates and spacing clusters so that cross-cluster sampling of individuals can occur could help ameliorate deficiencies in the detector layout. These results have application for a wide array of species and sampling methods (e.g., DNA sampling, camera trapping, mark-resight and search-encounter) and suggest that clustered sampling can significantly reduce the effort necessary to provide reliable estimates of population density across large spatial extents that previously would have been infeasible with nonclustered sampling designs.     

Synthesis, crystal structure and magnetic properties of cubane and linear trinuclear nickel complexes formed by 3,5-dichloro-2-hydroxybenzylaminoacetic acid

The reaction of Ni(ClO₄)₂·6H₂O with 3,5-dichloro-2-hydroxy-benzylaminoacetic acid (H₂dchaa), NaN₃ and triethylamine in methanol solution or water solution under solvothermal methods leads to the formation of two completely different Ni^II compounds: [HN(C₂H₅)₃]₈·[Ni₄(dchaa)₄(N₃)₄]₂ (1) and [HN(C₂H₅)₃]₂·[Ni₃(dchaa)₄(H₂O)₄]₂·(H₂O)₂ (2). The complexes 1 and 2 have been characterized by elemental analyses, IR spectra and single-crystal X-ray diffraction. Structure analyses reveal that complex 1 is a cubane cluster, while the complex 2 is a linear trinuclear cluster. The magnetic investigation shows that complexes 1 and 2 exhibit a ferromagnetic coupling between Ni^II ions. Ac susceptibilities of 1 and 2 reveal no frequency-dependent out-of-phase signals and the corresponding magnetic properties were discussed.