Long-term blue light exposure induces RGC-5 cell death in vitro: involvement of mitochondria-dependent apoptosis,oxidative stress,and MAPK signaling pathways

The mechanism of blue light-induced retinal ganglion cell (RGC) injury is poorly understood. In this study, we established a patented light-emitting diode-based system to study the effects of long-term blue light exposure under culture conditions on RGC-5 cells. Long-term blue light exposure significantly reduced cell viability in a time-dependent manner and induced apoptosis and necrosis in RGC-5 cells. Long-term blue light exposure marked an increase in the expression of Bax and active Caspase-3 (p17), which was accompanied by Bcl-2 down-regulation, and displayed features of the mitochondria-dependent apoptosis pathway. Blue light exposure also increased the generation of reactive oxygen species (ROS), and was a strong inducer of ROS-sensitive protein nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression. Moreover, blue light exposure constitutively activated p38 mitogen-activated protein kinases and c-Jun NH2-terminal kinase (JNK), as well as induced the phosphorylation of extracellular signal-regulated kinase in the early phase, in blue light-exposed RGC-5 cells. The protein expression of c-jun and c-fos was further enhanced after RGC-5 cells were exposed to blue light. Taken together, these findings indicated that blue light induced RGC-5 cell line death in dependence upon exposure duration. The potential mechanisms for this phenomenon might be via activated mitochondria-dependent apoptosis, increased ROS production and protein expressions of Nrf2 and HO-1, and activated JNK/p38 MAPK signaling pathways.     

近30年来青藏高原高寒草地NDVI动态变化对自然及人为因子的响应

草地生态系统是陆地生态系统的重要组成部分,在调节气候、水土保持、防风固沙、保护生物多样性等方面发挥着重要作用。青藏高原是全球海拔最高的独特地域单元,平均海拔超过4000 m,素有“世界第三极”之称,亦是我国重要的生态安全屏障,其对气候变化敏感且易受人类活动的影响,属于气候变化敏感区和生态脆弱带。近年来,由于气候变化和人类活动的不断加剧,青藏高原区域气候和环境发生了重大变化,气候变暖、水污染、草地退化和沙化等问题已严重阻碍了当地社会经济的可持续发展。高寒草地是青藏高原主要的植被类型,在气候变化和人类活动加剧的背景下,青藏高原高寒草地植被的动态变化受到人们的广泛关注。归一化植被指数（Normalized difference vegetation index, NDVI）因能有效地反映植被覆盖程度和生长状况而被广泛应用于植被动态的研究中。气温与降水被认为是影响青藏高原植被动态的主要气候因子,放牧强度与人口数量则是主要人为因子。因此,研究高寒草地植被对气候变化和人类活动的响应机制对预测未来草地变化有着重要的意义。基于青藏高原生长季草地的NDVI、气温、降水、放牧强度及人口数量等数据,在县区尺度上,采用趋势分析法探究了1982—2013年青藏高原143个县区生长季草地NDVI动态变化、气候变化及人类活动的变化,同时采用面板数据模型分析了32年来青藏高原143个县区气候、人为因子变化对草地NDVI变化的相对贡献。研究结果显示：（1）青藏高原高寒草地生长季NDVI总体呈增长趋势,草地植被生长状态呈现“整体改善、局部退化”趋势;（2）青藏高原生长季平均气温与降水量整体增加,气候呈现“暖湿化”趋势;（3）在长时间尺度上,气候因子主导了青藏高原高寒草地NDVI的变化,降雨和气温的增加促进草地NDVI的增加,放牧强度的持续增加则导致草地NDVI的减少。     

Fine-Scale Spatial Genetic Structure in Emmer Wheat and the Role of Population Range Position

The extent of spatial genetic structure (SGS) within plant populations depends on seed and pollen dispersal distance, breeding type, level of self-fertilization and effective plant density. Self-fertilizing species with gravity-dispersed seeds are expected to have both small effective population sizes and low pollen movement leading to high genetic structure. Higher SGS can be expected in more patchy and peripheral populations because of lower plant density and population sizes, and lower intensity of gene flow. We tested these predictions analyzing SGS in two core and two peripheral populations of predominantly self-fertilizing emmer wheat. Analysis of SGS with 11 nuclear microsatellites revealed (1) a negative linear relationship between kinship coefficients, calculated for pairs of individuals, and the logarithm of geographical distance between members of the pairs, in all studied populations; and (2) a significant autocorrelation for a distance up to 5 m (core populations) or 20 m (peripheral populations). Pollen flow, estimated from comparison of nuclear and chloroplast variation, was spatially limited, as was seed dispersal. Our results support a hypothesized relationship between SGS intensity and breeding system, the mode of seed dispersal and the population range position (core vs. periphery).     

用裂解气液色谱法鉴定昆虫包涵体病毒的初步研究

用Shimadzu GC-9A气相色谱仪和PyR-2A管式炉裂解器,对11株昆虫包涵体病毒进行了裂解气相色谱鉴定,通过对“指纹图”的分析,既可明显地区分GV、NPV和CPV包涵体病毒彼此间的差异,亦可较好的区别不同分离株间的异同,实验结果初步证明,用裂解气液色谱法分析、鉴定昆虫包涵体病毒是可行的。     

High uncertainties detected in the wetlands distribution of the Qinghai–Tibet Plateau based on multisource data

Landscape and Ecological Engineering - Twenty wetland-related data products (including remote sensing datasets, compilation datasets and model simulation datasets) were collected to evaluate the...     

L-asparaginase inhibits invasive and angiogenic activity and induces autophagy in ovarian cancer

Recent work identified L‐asparaginase (L‐ASP) as a putative therapeutic target for ovarian cancer. We suggest that L‐ASP, a dysregulator of glycosylation, would interrupt the local microenvironment, affecting the ovarian cancer cell—endothelial cell interaction and thus angiogenesis without cytotoxic effects. Ovarian cancer cell lines and human microvascular endothelial cells (HMVEC) were exposed to L‐ASP at physiologically attainable concentrations and subjected to analyses of endothelial tube formation, invasion, adhesion and the assessment of sialylated proteins involved in matrix‐associated and heterotypic cell adhesion. Marked reduction in HMVEC tube formation in vitro, HMVEC and ovarian cancer cell invasion, and heterotypic cell‐cell and cell‐matrix adhesion was observed (P < 0.05–0.0001). These effects were associated with reduced binding to ß1integrin, activation of FAK, and cell surface sialyl Lewis^X (sLe^x) expression. No reduction in HMVEC E‐selectin expression was seen consistent with the unidirectional inhibitory actions observed. L‐ASP concentrations were non‐toxic to either ovarian cancer or HMVEC lines in the time frame of the assays. However, early changes of autophagy were observed in both cell types with induction of ATG12, beclin‐1, and cleavage of LC‐3, indicating cell injury did occur. These data and the known mechanism of action of L‐ASP on glycosylation of nascent proteins suggest that L‐ASP reduces of ovarian cancer dissemination and progression through modification of its microenvironment. The reduction of ovarian cancer cell surface sLe^x inhibits interaction with HMVEC and thus HMVEC differentiation into tubes, inhibits interaction with the local matrix reducing invasive behaviour, and causes cell injury initiating autophagy in tumour and vascular cells.     

Constitution and quantity of lysis buffer alters outcome of reverse phase protein microarrays

Application of novel technology to clinical samples requires optimization of procedures. Reverse phase protein lysate arrays use femtomolar quantities of tissue lysate from clinical samples with which to profile biochemical events happening in the tumor. We analyzed the effects of different tissue solubilization buffers on frozen ovarian tumor samples in order to identify the system with the best signal intensity dynamic range, reproducibility, tissue solubility, and signal consistency. A modified RIPA-like buffer supplemented with DTT and SDS was deemed optimal.     

Translational Predictive Biomarker Analysis of the Phase 1b Sorafenib and Bevacizumab Study Expansion Cohort

Predictive biomarkers are needed to triage patients to the best therapy. We prospectively planned examination of sequential blood, biopsy, and functional imaging with which to confirm the mechanism and to identify potential predictive biomarkers in a phase Ib clinical trial expansion of patients with solid tumors receiving sorafenib/bevacizumab. The maximally tolerated doses of sorafenib at 200 mg twice daily with bevacizumab at 5 mg/kg every other week were given to biopsiable patients. Patients were randomized to receive either sorafenib or bevacizumab monotherapy for the first 28-day cycle with the second drug added with cycle 2. Biopsies, dynamic contrast-enhanced MRI, and fluorodeoxyglucose-proton emission tomography were done pre-therapy and at 2 and 6 weeks (2 weeks into combination therapy). Tumor and serum proteomics, Ras/Raf mutational analysis, and functional imaging results were examined individually and across the dataset to identify potential changes predictive of response to therapy and those that confirm the biochemical drug mechanism(s). Therapy with sorafenib/bevacizumab resulted in clinical benefit in 45% of this mixed solid tumor group. ERK activation and microvessel density were decreased with monotherapy treatment with sorafenib or bevacizumab, respectively; whereas a decreased signal over the group of total AKT, phospho(p)-VEGF receptor2, p-endothelial nitric-oxide synthase, b-RAF, and cleaved poly(ADP-ribose) polymerase was associated with earlier progression of disease. Tumor metabolic activity decreased in those patients with clinical benefits lasting longer than 4 months, and activity increased with progression of disease. Cleavage of caspase 3 and poly(ADP-ribose) polymerase was increased, and Ki67 expression decreased in patients with prolonged clinical benefits, consistent with decreased proliferation and increased apoptosis. The conglomerate analysis, incorporating pharmacodynamic and tumor biochemistry, demonstrated sorafenib/bevacizumab-targeted vascular activity in the tumor. Results suggest potential biomarkers for which changes, as a group, during early therapeutic exposure may predict clinical benefit.Sorafenib and bevacizumab have demonstrated clinical utility as single agents or in combination with chemotherapy for solid tumors. Sorafenib, initially developed as a c-Raf kinase inhibitor, also has potent inhibitory activity against the vascular endothelial growth factor receptor-2 (VEGFR2).¹ Clinical activity has been shown for bevacizumab, the humanized neutralizing monoclonal antibody against VEGF, also alone and in chemotherapy combinations (1–5). The role of combining two agents with overlapping target biology had not yet been studied.We tested the clinical hypothesis that signal interruption at collaborative pathway points, both vertical and horizontal interactions, may yield equal or greater effect than the agents in isolation in a phase I trial combining bevacizumab and sorafenib ({"type":"clinical-trial","attrs":{"text":"NCT00095459","term_id":"NCT00095459"}}NCT00095459), and we now report the translational analyses (6). Sorafenib was selected for its ability to target both receptor and cytosolic kinases important in a variety of activated cells in the tumor microenvironment, including stromal, endothelial, and malignant cells. Because such kinase inhibitor treatment has been shown to up-regulate production of proangiogenic cytokines, we added bevacizumab to reduce VEGF ligand availability and augment inhibition of endothelial cells. We observed the clinical benefit, including partial response and prolonged disease stabilization, using attenuated doses of the individual agents as determined by safety assessments during the trial; partial response or disease stabilization of at least 4 months occurred in 59% of the daily sorafenib cohort and in 55% of those on the intermittent, 5 of 7 days, sorafenib schedule (6, 7). These benefits lasted up to 37+ months with over 25% of patients receiving 12 or more months of therapy. The trial prospectively planned comprehensive translational assessment using a randomized drug addition design (Fig. 1A) to evaluate individual drug target specificity and combination drug effects to identify potential predictive biomarkers to examine in the ongoing phase II study of sorafenib/bevacizumab in ovarian cancer.Open in a separate windowFig. 1.Treatment schema (A) and Consort diagram (B).Predictive biomarkers are increasingly important for the advancement of targeted therapies. Such knowledge should allow more effective triage of patients to interventions more likely to provide clinical benefit. Biomarkers that predict drug response may consist of direct measures of activity, such as modulation of biochemical signals in the tumor (8) or those that yield pharmacodynamic measures, such as functional imaging (9, 10). Changes in metabolic activity and/or blood flow using dynamic imaging may fall into both categories with decreased glucose uptake due to reduced glucose delivery and/or reduced glucose metabolism and altered vascular permeability in response to attenuation of the VEGF drive. Aggregate analysis of these varied translational measures may yield a more detailed view of the cancer and the drug combination, allowing broader dissection into potential predictive biomarkers. Linking modulation of activity with clinical benefit is a first step in validating prospective biomarkers.We designed a novel drug administration schema from which to examine the contribution of both sorafenib and bevacizumab on the modulation of tumor and the tumor microenvironment behavior. The biochemical and imaging data demonstrate changes consistent with alteration of tumor vascularity, demonstrate direct association of target effect with clinical outcome across solid tumor types, and confirm the benefit of complementary pathway targeting. Reduction in blood flow, up-regulation of cytokine production, and inhibition of a set of anti-apoptotic anti-proliferative signaling events together may define potentially predictive changes to examine in early drug administration in subsequent trials.     

Microglial replacement in the aged brain restricts neuroinflammation following intracerebral hemorrhage

Aged microglia display augmented inflammatory activity after neural injury. Although aging is a risk factor for poor outcome after brain insults, the precise impact of aging-related alterations in microglia on neural injury remains poorly understood. Microglia can be eliminated via pharmacological inhibition of the colony–stimulating factor 1 receptor (CSF1R). Upon withdrawal of CSF1R inhibitors, microglia rapidly repopulate the entire brain, leading to replacement of the microglial compartment. In this study, we investigated the impact of microglial replacement in the aged brain on neural injury using a mouse model of intracerebral hemorrhage (ICH) induced by collagenase injection. We found that replacement of microglia in the aged brain reduced neurological deficits and brain edema after ICH. Microglial replacement-induced attenuation of ICH injury was accompanied with alleviated blood-brain barrier disruption and leukocyte infiltration. Notably, newly repopulated microglia had reduced expression of IL-1β, TNF-α and CD86, and upregulation of CD206 in response to ICH. Our findings suggest that replacement of microglia in the aged brain restricts neuroinflammation and brain injury following ICH.Subject terms: Neuroimmunology, Cognitive ageing     

Differential response of oilseed rape (Brassica napus L.) cultivars to low boron supply

Three experiments were carried out on an alluvial sandy loam (Udifluvent) at Tonglu, Zhejiang Province, P.R. China from 1992 to 1995, to determine the genotypic range in boron (B) efficiency of 16 oilseed rape (Brassica napus L.) cultivars, to identify the B-efficient cultivars and to identify specific responses which can be utilised for selection in a breeding program. The 16 cultivars which included high-quality and conventional types differed significantly in survival, plant height, leaf area, shoot dry weight and seed yield; however, the ranking of the cultivars for their seed yield or other plant traits differed with B treatment. With severe B deficiency (CaCl2 extractable B < 0.26 mg/kg) and no boron applied, none of the cultivars exhibited significant B efficiency, with seed yield <300 kg/ha. With moderate B deficiency (CaCl2 extractable B 0.34 mg/kg or 0.17 kg B/ha applied), seed yield varied significantly among the cultivars from 397 to 1889 kg/ha in year 1 and from 616 to 1260 kg/ha in year 3. Zhongyou 821 and 92-13 were the most B-efficient and Wanyou 324, Huashuang 2 and Su 2051 were the most B-inefficient cultivars under moderate B deficiency. Significant differences were found among the cultivars in leaf B concentration; however, there was no close relationship between leaf B concentration and seed yield responses to B of oilseed cultivars. Of all the growth parameters measured, leaf area was the early indicator best correlated with subsequent seed yield and may be useful for evaluating the response of cultivars to low B supply. Contrary to current opinion, it was also found that high-quality oilseed rape cultivars were not all sensitive to low B supply nor were all conventional cultivars B-efficient.