Specificity of Collybistin-Phosphoinositide Interactions: IMPACT OF THE INDIVIDUAL PROTEIN DOMAINS*

The regulatory protein collybistin (CB) recruits the receptor-scaffolding protein gephyrin to mammalian inhibitory glycinergic and GABAergic postsynaptic membranes in nerve cells. CB is tethered to the membrane via phosphoinositides. We developed an in vitro assay based on solid-supported 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine membranes doped with different phosphoinositides on silicon/silicon dioxide substrates to quantify the binding of various CB2 constructs using reflectometric interference spectroscopy. Based on adsorption isotherms, we obtained dissociation constants and binding capacities of the membranes. Our results show that full-length CB2 harboring the N-terminal Src homology 3 (SH3) domain (CB2_SH3+) adopts a closed and autoinhibited conformation that largely prevents membrane binding. This autoinhibition is relieved upon introduction of the W24A/E262A mutation, which conformationally “opens” CB2_SH3+ and allows the pleckstrin homology domain to properly bind lipids depending on the phosphoinositide species with a preference for phosphatidylinositol 3-monophosphate and phosphatidylinositol 4-monophosphate. This type of membrane tethering under the control of the release of the SH3 domain of CB is essential for regulating gephyrin clustering.     

Comparison of Southern Appalachian high-elevation outcrop plant communities with their Northern Appalachian counterparts

Abstract. Southern Appalachian high-elevation outcrops harbour six regionally rare Northern Appalachian taxa usually considered relicts of a Pleistocene alpine flora. For five of the six taxa, minimum elevation in the south was 367–1113 m higher than in the north. While habitats compared between the two regions share only 9% of their total flora, individual plots had up to 70% of their species occurring in the opposite region. The northern affinity of southern outcrops increased with elevation, slope steepness, soil Cu, B and SO₄ and decreased with potential solar radiation and soil Na. As a result, communities above 1600 m on felsic bedrock, and above 1350 m on mafic bedrock, were most northern in composition. Northern affinity of southern outcrops also increased with latitude, which may partly result from closer geographic proximity to past communities that provided progenitors for the current northern flora. Northern treeless habitats increased in southern affinity with increased slope steepness, perennial seepage, vegetation height, shade, soil pH, Al, Mn, Na and decreased elevation and organic matter. As a result, northern outcrop communities below treeline were most similar to those on southern outcrops. This suggests that southern outcrop vegetation may be more similar to Pleistocene outcrop vegetation than to Pleistocene alpine vegetation. Partial constrained ordination showed that while compositional differences between the Northern and Southern Appalachian habitats were largely explained by environmental differences, there was a significant component of residual variation explained by north or south position that was unrelated to environment. These residual compositional differences may result from historical influences on community structure involving stochastic extinction and colonization processes.     

Comparative Ploidy Studies Using Cytological and Paraffin Section Preparations

The ploidy profiles of benign and malignant tumours can be obtained using image analysis. However, the results of ploidy studies have varied according to the type of specimen used. We compared the ploidy profiles of paraffin embedded thin sections, cytospin preparations of disaggregated cells, and cytological smears from the same specimen as defined by image analysis. Ten benign breast lesions, 10 breast carcinomas and 10 malignant melanomas were investigated in this way. Preparations stained by the Feulgen technique were examined using the MD20 video image analysis densitometry system. Ploidy profiles were obtained by measuring the integrated optical density of at least 200 nuclei. By paying proper attention to the quality of fixation and presence or absence of cytoplasm around cells, comparable results were found for all preparations in each case. We therefore conclude that if careful attention is paid to the technical quality of the material, reliable ploidy results can be obtained by image analysis.     

The Alpha and Omega of Galactosylceramides in T Cell Immune Function

Glycosphingolipids are a subgroup of glycolipids that contain an amino alcohol sphingoid base linked to sugars. They are found in the membranes of cells ranging from bacteria to vertebrates. This group of lipids is known to stimulate the immune system through activation of a type of white blood cell known as natural killer T cell (NKT cell). Here we summarize the extensive research that has been done to identify the structures of natural glycolipids that stimulate NKT cells and to determine how these antigens are recognized. We also review studies designed to understand how glycolipid variants, both natural and synthetic, can alter the responses of NKT cells, leading to dramatic changes in the global immune response.     

DIA-Based Proteomics Identifies IDH2 as a Targetable Regulator of Acquired Drug Resistance in Chronic Myeloid Leukemia

Drug resistance is a critical obstacle to effective treatment in patients with chronic myeloid leukemia. To understand the underlying resistance mechanisms in response to imatinib mesylate (IMA) and adriamycin (ADR), the parental K562 cells were treated with low doses of IMA or ADR for 2 months to generate derivative cells with mild, intermediate, and severe resistance to the drugs as defined by their increasing resistance index. PulseDIA-based (DIA [data-independent acquisition]) quantitative proteomics was then employed to reveal the proteome changes in these resistant cells. In total, 7082 proteins from 98,232 peptides were identified and quantified from the dataset using four DIA software tools including OpenSWATH, Spectronaut, DIA-NN, and EncyclopeDIA. Sirtuin signaling pathway was found to be significantly enriched in both ADR-resistant and IMA-resistant K562 cells. In particular, isocitrate dehydrogenase (NADP(+)) 2 was identified as a potential drug target correlated with the drug resistance phenotype, and its inhibition by the antagonist AGI-6780 reversed the acquired resistance in K562 cells to either ADR or IMA. Together, our study has implicated isocitrate dehydrogenase (NADP(+)) 2 as a potential target that can be therapeutically leveraged to alleviate the drug resistance in K562 cells when treated with IMA and ADR.     

Effect of pea bacterial blight (Pseudomonas syringae pv. pisi) on the yield of spring sown combining peas (Pisum sativum)

Field trials to examine the effect of pea bacterial blight (Pseudomonas syringae pv. pisi) (Psp) on the yield of combining peas were carried out at five sites (HRI Wellesbourne, ADAS Rosemaund, ADAS Terrington, PGRO, SASA East Craigs) in the UK in 1990, 1991 and 1992. Healthy seed, cv. Solara, and seed naturally infected with Psp Race 2 was sown in large plots (c. 200 m²) in the open or under nets to prevent bird damage by pigeons. Despite relatively low disease severity levels (< 15% leaf area) and separation by at least 12 m of cv. Consort (resistant to Race 2) between plots there was considerable spread of disease into plots sown with healthy seed. Regression analysis demonstrated a significant relationship between yield and disease. Of the disease measurements examined, disease severity on the leaves (stipules) at growth stage 208 was found to be the best predictor of yield. A model containing environmental and bird damage terms, in addition to disease, suggested that a yield loss of 0.98 t/ha would be expected for a disease severity score of 1, equivalent to 5% leaf area affected, at growth stage 208.     

Biodiesel preparation from microalgae lipid by two-step lipase catalysis

To overcome the high energy-consuming process of microalgae drying, a two-step lipase catalysis technique for the preparation of biodiesel from microalgae lipid of Chlorella spp. was developed. In the first step, free fatty acids (FAAs) and triacylglycerols (TAGs) are released after cell disruption and extracted, while the TAGs were hydrolysed by free lipase in aqueous phase. In the second step, FAAs were esterified with ethanol in the catalysis of free suspended lipase. The maximum rate of hydrolysis and esterification was 93.6% and 91.3%, respectively. The effects of reaction parameters, such as reaction time, enzyme amount, water content and molar ratio of lipid to ethanol on hydrolysis or esterification, were investigated. The results indicated that two-step reaction process (hydrolyse esterify) for biodiesel production were feasible.     

重庆市生态风险预警等级划分及演化趋势模拟

生态风险预警等级评估和演化趋势模拟，可为生态风险管理提供可靠的辅助决策。以重庆市为研究对象，基于驱动力-压力-状态-影响-响应模型，构建重庆市生态风险预警指标体系，采用正态云模型和集对分析法，定量分析重庆市生态风险时空分异特征和演化趋势。研究结果表明：（1）2013-2019年，重庆市生态风险值呈"上升-下降"的波动趋势，综合生态风险隶属于重警等级，生态风险综合值从0.295下降到0.278，生态环境逐年好转；（2）重庆市生态风险有下降、不变、先上升后降低、先降低后上升以及一直升高5种演化趋势，分别占比39%、16%、5%、21%、24%；（3）重庆市生态风险转移分为两个方向，2013-2016年生态风险空间分异性增大，中警、轻警和无警风险等级不断向东北、东南和西部四周分散转移；2016-2019年生态风险分布格局变化较小，重警风险区在东部聚集；（4）演化趋势模拟结果表明，未来重庆市生态风险降低的区县有13个，占比34%，生态环境有向好发展的趋势；生态风险上升的区县有25个，占比66%，生态环境会有所恶化，但是恶化程度较低。将生态风险等级划分与预警演化趋势相结合，能为城市生态风险管理提供科学依据。     

基于MODIS NDVI的长江中游区域植被动态及与气候因子的关系

基于1999-2015年的MODIS NDVI时间序列遥感数据，应用趋势分析、变异系数、重标极差分析和偏相关分析等方法，分析了长江中游的植被时空变化特征及其与气象因子的关系。结果表明，长江中游地区NDVI均值总体上呈上升趋势（从0.72增加到0.80）。从空间分布来看，NDVI低值区域（0.1-0.5）占1.40%，高值区域（>0.7）占87.15%；NDVI空间格局呈"西高东低、北高南低"的分布特征，低值区域表现为以三省省会城市为中心向外辐射。Hurst指数显示，研究区大部分区域（60.54%）的NDVI变化趋势具有不确定性，持续性改善区域（34.78%）主要分布在西部山地区，持续性退化区域（3.26%）主要分布在人类活动频繁的较发达城市区域。在年际尺度上，研究区NDVI与各气象因子关系均不显著；月际尺度上，NDVI与降水、相对湿度和日照时数显著相关，降水和日照时数有明显的时滞性。区域内NDVI动态趋势以不确定性发展为主，城市群周边NDVI呈现持续退化的区域应该引起关注。     

分泌型磷脂酶PLA2G5的生物学功能及其抑制剂研究进展

分泌型磷脂酶PLA2G5属于磷脂酶A2超家族的一员,在免疫细胞和非免疫细胞中均有表达.研究表明,PLA2G5参与生物学事件的发生发展,在特定的病理条件下具有诱导作用.本文简要阐述了PLA2G5的来源、结构特征、生物学功能和在疾病中的作用,以及现有或潜在的PLA2G5抑制剂,以期探索基于PLA2G5的治疗新靶标.