Chaperone gp96-independent inhibition of endotoxin response by chaperone-based peptide inhibitors

HSP90 chaperones a large number of proteins, and it plays essential roles in multiple signaling pathways to maintain protein homeostasis in the cytosol. In addition, HSP90 has been implicated in mediating recognition of lipopolysaccharide (LPS). However, no pharmacologic agents have been developed to interrogate this pathway. Herein we demonstrate that a peptide-based inhibitor that was previously reported to inhibit the master Toll-like receptor-chaperone gp96, an endoplasmic reticulum paralog of HSP90, in fact blocks HSP90-LPS interaction. It inhibited the binding of LPS to the cell surface of both wild type and gp96-null cells and thereby abrogated the cellular response to LPS but not to other Toll-like receptor ligands. We also generated a series of peptide derivatives (named peptide inhibitors of endotoxin responsiveness (PIERs)) from the N-terminal helix structure of HSP90 and demonstrated their effectiveness in blocking LPS activity. PIER inhibition of LPS signaling was partially reversed by CD14 expression. Moreover, we found that a cell-permeable PIER abrogated HSP90 function and caused degradation of multiple known HSP90 client proteins in cancer cells. Thus, targeting HSP90 is a promising modality for treatment of both LPS-mediated pathology and cancer.     

人热休克蛋白gp96基因的克隆及其在大肠杆菌中的表达与纯化

热休克蛋白gp96是热休克蛋白90家族成员,能够引起非特异性和特异性免疫反应。得到大量高纯度的蛋白质是研究开发gp96的关键。然而重组的gp96容易在E．coli中降解,并在一定条件下形成多聚体。实验先将人gp96基因克隆到pET-30a载体上并在E．coli Blstar中表达,再经过亲和层析、阴离子交换、分子筛分别纯化gp96。最终去掉了大部分的降解片段和多聚体,得到一定量的可溶性gp96,为进一步研究其结构和功能打下一定的基础。     

灵芝菌丝发酵提取液减肥作用的初步探讨

目的:对灵芝菌丝发酵提取液的减肥作用进行初步研究.方法:用高营养饲料建立营养性肥胖小鼠模型后,治疗组用灵芝菌丝发酵提取液灌胃,对照组用无菌生理盐水,观察体重变化.结果:灵芝菌丝发酵提取液能明显降低肥胖小鼠体重.讨论:灵芝菌丝发酵提取液具有减肥、降低体重作用.     

Isolation,Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson’s diseae. Study of the biological processes involved in physiological functions and vulnerability and death of these neurons is imparative to understanding the underlying causes and unraveling the cure for this common neurodegenerative disorder. Primary cultures of mesDA neurons provide a tool for investigation of the molecular, biochemical and electrophysiological properties, in order to understand the development, long-term survival and degeneration of these neurons during the course of disease. Here we present a detailed method for the isolation, culturing and maintenance of midbrain dopaminergic neurons from E12.5 mouse (or E14.5 rat) embryos. Optimized cell culture conditions in this protocol result in presence of axonal and dendritic projections, synaptic connections and other neuronal morphological properties, which make the cultures suitable for study of the physiological, cell biological and molecular characteristics of this neuronal population.     

Development of seawater adaptation in pink salmon,Oncorhynchus gorbuscha,during downstream migration: relationships to temperature and residual yolk

Synopsis Seawater adaptability of wild pink salmon, Oncorhynchus gorbuscha, alevins was investigated using 96 h LC50 test, muscle sodium level and weight loss tests. Five temperature (2, 6, 9, 13 and 16°C) and two yolk groups were analysed during the beginning, the middle and the end of downstream migration. Both temperature and amount of residual yolk affected seawater adaptability. The computation of muscle sodium levels and weight loss after a short term (3–12 h) seawater challenge test may be a convenient method to estimate the seawater adaptability of fish too small to allow analysis of blood sodium concentration.     

Ageing and longevity in the Decapoda (Crustacea): A review

Ageing and longevity is a neglected field of crustacean biology. Information on longevity is available for less than 2% of the extant species of the Decapoda. Maximum ages reliably determined range from 40 days to 72 years corresponding to a life span difference of a factor of 650. The shortest-lived decapods are planktonic dendrobranchiate shrimps, and particularly long-lived species with life spans of decades are found in the Astacidea. Most decapods seem to live for 1-10 years. High geographical latitude, the deep sea and freshwater caves promote longevity. The majority of the Decapoda is indeterminately growing and presumably characterized by negligible senescence. The adults of the determinately growing decapods like some brachyuran crabs suffer from mechanical senescence and are unable to regenerate lost appendages. The decapod crustaceans have developed many effective anti-ageing mechanisms including moulting, detoxification of free radicals, removal of cellular waste, renewal of tissues by life-long stem cell activity, regeneration of appendages, detoxification of environmental pollutants and isolation of pathogens and diseased tissue areas by melanisation and encapsulation. Age related diseases including cancer are virtually unknown. The present compilation of data on longevity and senescence in decapods is the first one that covers the whole spectrum of a higher invertebrate taxon. It is hoped to provide an interesting source of information for carcinologists and biogerontologists. Further improvement of knowledge on ageing and longevity in the Decapoda would be beneficial for crustacean aquaculture, fisheries and ecological modelling. Some decapods even have good potential to become models for general ageing research.     

AIMP3 inhibits cell growth and metastasis of lung adenocarcinoma through activating a miR-96-5p-AIMP3-p53 axis

Aminoacyl-tRNA synthetase-interacting multifunctional protein-3 (AIMP3) is a tumour suppressor, however, the roles of AIMP3 in non-small cell lung cancer (NSCLC) are not explored yet. Here, we reported that AIMP3 significantly inhibited the cell growth and metastasis of NSCLC (lung adenocarcinoma) in vitro and in vivo. We have firstly identified that AIMP3 was down-regulated in human NSCLC tissues compared with adjacent normal lung tissues using immunohistochemistry and western blot assays. Overexpression of AIMP3 markedly suppressed the proliferation and migration of cancer cells in a p53-dependent manner. Furthermore, we observed that AIMP3 significantly suppressed tumour growth and metastasis of A549 cells in xenograft nude mice. Mechanically, we identified that AIMP3 was a direct target of miR-96-5p, and we also observed that there was a negative correlation between AIMP3 and miR-96-5p expression in paired NSCLC clinic samples. Ectopic miR-96-5p expression promoted the proliferation and migration of cancer cells in vitro and tumour growth and metastasis in vivo which partially depended on AIMP3. Taken together, our results demonstrated that the axis of miR-96-5p-AIMP3-p53 played an important role in lung adenocarcinoma, which may provide a new strategy for the diagnosis and treatment of NSCLC.     

高剂量热休克蛋白gp96通过激活调节性T细胞预防1型糖尿病

旨在以非肥胖糖尿病(Non-obese diabetic,NOD)小鼠为动物模型,研究高剂量昆虫细胞表达的重组热休克蛋白gp96(Recombinant gp96,rgp96)对1型糖尿病(Type 1 diabetes,T1D)的预防作用。以高剂量rgp96免疫NOD小鼠,用血糖仪监测小鼠血糖值,用流式细胞术检测小鼠脾脏CD4~+CD25~+Foxp3~+调节性T细胞(Regulatory T cells,Tregs)亚群频率,然后用一系列体外实验探究高剂量rgp96对Tregs的影响。结果显示高剂量rgp96免疫有效地预防或延缓小鼠T1D发病,免疫诱导Tregs数量明显增加。体外实验发现rgp96蛋白促进Tregs增殖,诱导Foxp3表达上调和IL-10分泌增加。研究结果为开发基于rgp96的新型T1D预防和治疗性疫苗提供了依据。     

A federated database of taxon pages in the Italian Biodiversity Network

Primary biodiversity data are often organised in federated databases in the framework of big international projects and initiatives. Other data are normally organised into isolated resources: ecological, morphological and morpho-anatomic databases, checklists, archives of digital identification tools, etc. These data, which do not refer specifically to one specimen or observation, but to a taxon, can be addressed to as “taxon-related data”. In the framework of the Italian Biodiversity Network, taxon-related data have been aggregated by using a federated approach, with the creation of a network of data providers, which required the development of a communication protocol, the species-related data (SRD) protocol and of a simple data format (the SRD format). The SRD federated database aggregates currently ca. 70,000 taxon pages for several groups of organisms: vascular plants, lichens, mosses, algae, freshwater fishes and butterflies. All these resources can be queried by using one simple interface in the web portal of the Italian Biodiversity Network (http://www.naturaitalia.it/nnb/).