烟草甲clip丝氨酸蛋白酶基因的克隆及其对外源激素和免疫胁迫的表达响应(英文)

【目的】作为细胞外信号级联通路的重要组成部分,含有clip结构域的丝氨酸蛋白酶(clip-domain serine proteases, CLIPs)在昆虫发育和先天免疫过程中起着重要作用。本研究旨在克隆烟草甲Lasioderma serricorne CLIP基因,解析其在烟草甲不同发育阶段和幼虫不同组织中的表达模式,分析其在外源激素20-羟基蜕皮酮(20E)和免疫胁迫后的表达特征,为进一步研究其生理功能奠定基础。【方法】采用RT-PCR技术克隆获得烟草甲两个CLIPs基因(LsCLIP1和LsCLIP2)全长cDNA序列,并利用生物信息学软件预测其编码蛋白的结构和特征,利用MEGA 6.06构建昆虫CLIPs系统发育树;利用实时荧光定量PCR(quantitative real-time PCR, qPCR)研究这两个基因在不同发育阶段［低龄幼虫(卵孵化后24 h内)、高龄幼虫(4龄以上)、蛹(化蛹后48 h以上)、早期成虫(化蛹后24 h内)和晚期成虫(化蛹后7 d)］、5龄幼虫不同组织(表皮、脂肪体、肠道和剩余组织)中以及注射20E(120 ng/幼虫)和来源于大肠杆菌Escherichia coli和金黄色葡萄球菌Staphylococcus aureus的肽聚糖(0.2 μL)后4龄幼虫中的表达模式。【结果】克隆获得烟草甲LsCLIP1和LsCLIP2基因的cDNA全序列,其开放阅读框长度均为1 194 bp,编码397个氨基酸。序列分析显示,其氨基酸序列各自具有一个clip结构域和胰蛋白酶结构域。系统发育分析表明,CLIP1和CLIP2都属于subfamily C CLIPs。qPCR结果表明,LsCLIP1和LsCLIP2基因在所检测的各发育阶段和幼虫各组织中均有表达,分别尤以蛹期和表皮中表达量最高;经20E和肽聚糖诱导后,烟草甲幼虫体内LsCLIP1和LsCLIP2基因的表达量明显提高。【结论】推测LsCLIP1和LsCLIP2可能参与了烟草甲的蜕皮发育和对免疫胁迫的应激响应。本研究将为后续研究昆虫CLIPs的分子调控提供参考。     

Epstein–Barr virus noncoding RNAs from the extracellular vesicles of nasopharyngeal carcinoma (NPC) cells promote angiogenesis via TLR3/RIG-I-mediated VCAM-1 expression

Viral noncoding RNAs (Epstein–Barr virus-encoded RNAs, EBERs) are believed to play a critical role in the progression of lymphoma and nasopharyngeal carcinoma (NPC). However, the accurate mechanisms accounting for their oncogenic function have not been elucidated, especially in terms of interaction between tumor cells and mesenchymal cells. Here, we report that, in addition to NPC cells, EBERs are also found in endothelial cells in Epstein–Barr virus (EBV)-infected NPC parenchymal tissues, which implicates NPC-derived extracellular vesicles (EVs) in transmitting EBERs to endothelial cells. In support of this hypothesis, we first ascertained if EBERs could be transferred to endothelial cells via EVs isolated from NPC culture supernatant. Then, we clarified that EVs-derived EBERs could promote angiogenesis through stimulation of VCAM-1 expression. Finally, we explored the involvement of EBER recognition by TLR3 and RIG-I in NPC angiogenesis. Our observations collectively illustrate the significance and mechanism of EVs-derived EBERs in angiogenesis and underlie the interaction mechanisms between EBV-infected NPC cells and the tumor microenvironment.     

Repellent action and contact toxicity mechanisms of the essential oil extracted from Chinese chive against Plutella xylostella larvae

Botanical pesticides play increasingly important roles in the control of agricultural pests. In this study, the insecticidal effects, specifically the repellent action and contact toxicity, of the essential oil extracted from Chinese chive (EOC) against Plutella xylostella larvae were confirmed. The mechanisms of repellent’s action were studied using electroantennograms (EAGs), and the effects on glutathione S‐transferase (GST), carboxylesterase (CarE), and acetyl cholinesterase were investigated after EOC treatments. The EOC affected the EAG results and inhibited the activities of GST and CarE in treated P. xylostella larvae, which could explain its insecticidal effects. And, four pyrazines showed greater repellent activities than that of the EOC, which was confirmed as the main active compounds of EOC.     

An evaluation of transferability of ecological niche models

Ecological niche modeling (ENM) is used widely to study species’ geographic distributions. ENM applications frequently involve transferring models calibrated with environmental data from one region to other regions or times that may include novel environmental conditions. When novel conditions are present, transferability implies extrapolation, whereas, in absence of such conditions, transferability is an interpolation step only. We evaluated transferability of models produced using 11 ENM algorithms from the perspective of interpolation and extrapolation in a virtual species framework. We defined fundamental niches and potential distributions of 16 virtual species distributed across Eurasia. To simulate real situations of incomplete understanding of species’ distribution or existing fundamental niche (environmental conditions suitable for the species contained in the study area; N* _F ), we divided Eurasia into six regions and used 1–5 regions for model calibration and the rest for model evaluation. The models produced with the 11 ENM algorithms were evaluated in environmental space, to complement the traditional geographic evaluation of models. None of the algorithms accurately estimated the existing fundamental niche (N* _F ) given one region in calibration, and model evaluation scores decreased as the novelty of the environments in the evaluation regions increased. Thus, we recommend quantifying environmental similarity between calibration and transfer regions prior to model transfer, providing an avenue for assessing uncertainty of model transferability. Different algorithms had different sensitivity to completeness of knowledge of N* _F , with implications for algorithm selection. If the goal is to reconstruct fundamental niches, users should choose algorithms with limited extrapolation when N* _F is well known, or choose algorithms with increased extrapolation when N* _F is poorly known. Our assessment can inform applications of ecological niche modeling transference to anticipate species invasions into novel areas, disease emergence in new regions, and forecasts of species distributions under future climate conditions.     

Oleuropein induces apoptosis via abrogating NF-κB activation cascade in estrogen receptor–negative breast cancer cells

Oleuropein is one of the most abundant phenolic compounds found in olives. Epidemiological studies have indicated that an increasing intake of olive oil can significantly reduce the risk of breast cancer. However, the potential effect(s) of oleuropein on estrogen receptor (ER)-negative breast cancer is not fully understood. This study aims to understand the anticancer effects and underlying mechanism(s) of oleuropein on ER-negative breast cancer cells in vitro. The effect of oleuropein on the viability of breast cancer cell lines was examined by mitochondrial dye-uptake assay, apoptosis by flow cytometric analysis, nuclear factor-κB (NF-κB) activation by DNA binding/reporter assays and protein expression by Western blot analysis. In the present report, thiazolyl blue tetrazolium bromide assay results indicated that oleuropein inhibited the viability of breast cancer cells, and its effects were more pronounced on MDA-MB-231 as compared with MCF-7 cells. It was further found that oleuropein increased the level of reactive oxygen species and also significantly inhibited cellular migration and invasion. In addition, the activation of NF-κB was abrogated as demonstrated by Western blot analysis, NF-κB-DNA binding, and luciferase assays. Overall, the data indicates that oleuropein can induce substantial apoptosis via modulating NF-κB activation cascade in breast cancer cells.     

A seascape genetic analysis of a stress-tolerant coral species along the Western Australian coast

Coral Reefs - Genetic diversity and connectivity are key factors in determining a population’s resilience to future disturbance. This is especially relevant to corals, which are in global...     

Effects of calpain inhibitor on the apoptosis of hepatic stellate cells induced by calcium ionophore A23187

We previously showed that changes in calcium concentrations were related to cell apoptosis in vitro. The endoplasmic reticulum (ER) is the main component of calcium storage and signal transduction, and disrupting the balance of intracellular Ca²⁺ can cause endoplasmic reticulum stress (ERS). In this process, the ER releases stored Ca ²⁺ into the cytoplasm and activates calpain-2. To further investigate the effect of calpain in hepatic stellate cells (HSCs), in the current study, we examine the effect of N-acetyl-leu-leu-norleucinal (ALLN) on apoptosis resulting from calcium ionophore A23187–induced ERS. Our findings indicate that calpain inhibition reduces calcium ionophore A23187–induced apoptosis of HSCs and decreases the expression of ER stress proteins that may be related to the calpain/caspase signaling pathway.