亚成体大熊猫肠道真菌多样性

【目的】分析5只亚成体大熊猫肠道真菌的多样性。【方法】采用基于ITS基因的RFLP(Restriction Fragment Length Polymorphism)方法,对肠道真菌总DNA进行ITS-RFLP分析,构建ITS克隆文库,然后用HhaI、HaeIII进行酶切指纹图谱分析,测序并绘制系统进化树。【结果】研究表明,5只亚成体大熊猫肠道真菌主要由Ascomycota(平均占46.24%)、Basidiomycota(平均占15.79%)2个门和一些未分类(平均占29.14%)、未培养(平均占8.83%)的真菌。其中,Ascomycota主要以Saccharomycetes(平均占63.74%)和Dothideomycetes(平均占35.91%)2个纲为主;Basidiomycota主要以Tremellomycetes(平均占65.80%)和Microbotryomycetes(平均占33.15%)2个纲为主。而这4个纲分别则主要以Candida、Debaryomyces;Pleosporales、Myriangium;Cystofilobasidium、Trichosporon;Leucosporidium、Leucosporidiella八个菌属为主,各样品中所占比例不等。【结论】亚成体大熊猫肠道内存在一定比例的真菌菌群,且ITS-RFLP技术能够很好地对其多样性进行分析。真菌的发现扩大了我们对大熊猫肠道微生物结构的了解,同时也有助于我们进一步研究真菌能否帮助大熊猫消化高纤维素食物。     

Accuracy and repeatability of moose (<Emphasis Type="Italic">Alces alces</Emphasis>) age as estimated from dental cement layers

Optimal research and management of species with age structure often depends on estimates of age-specific population parameters, which in turn depends on reliable methods for age determination. By counting annuli in the cementum of incisor root tips from 51 known-age moose (Alces alces) between 1 and 12 years old, we examined the variation in accuracy and repeatability of age estimates provided by three research technicians with different experiences of aging moose. The most experienced technician estimated the moose age correctly in up to 90% of the cases, while the technician with no prior experience estimated age correctly in up to 73% of the cases. The medium-experienced technician achieved a lower accuracy (up to 53%), indicating that experience alone is not sufficient if the basic training or lack of routine checks against other colleagues or a known-age material are not undertaken. The percentage moose aged within ±1 year from correct age was significantly higher in all technicians (94–98%). After the generally high accuracy, we also found high repeatability (0.980–0.994) within technicians. We conclude that this method of age-determination provides reliable estimates that can be used by management and research to gain information about age-specific patterns in moose populations. However, to obtain estimates of high accuracy the technicians should be well trained, have gained the necessary experience, and most preferably, have access to a sample of teeth from known-age moose.     

Cloning,expression, and characterization of thermostable Manganese superoxide dismutase from <Emphasis Type="Italic">Thermoascus aurantiacus</Emphasis> var. <Emphasis Type="Italic">levisporus</Emphasis>

A superoxide dismutase (SOD) gene of Thermoascus aurantiacus var. levisporus, a thermophilic fungus, was cloned, sequenced, and expressed in Pichia pastoris and its gene product was characterized. The coding sequence predicted a 231 residues protein with a unique 35 amino acids extension at the N-terminus indicating a mitochondrial-targeting sequence. The content of Mn was 2.46 μg/mg of protein and Fe was not detected in the purified enzyme. The enzyme was found to be inhibited by NaN₃, but not by KCN or H₂O₂. These results suggested that the SOD in Thermoascus aurantiacus var. levisporus was the manganese superoxide dismutase type. In comparison with other MnSODs, all manganese-binding sites were also conserved in the sequence (H88, H136, D222, H226). The molecular mass of a single band of the enzyme was estimated to be 21.7 kDa. The protein was expressed in tetramer form with molecular weight of 68.0 kDa. The activity of purified protein was 2,324 U/mg. The optimum temperature of the enzyme was 55°C and it exhibited maximal activity at pH 7.5. The enzyme was thermostable at 50 and 60°C and the half-life at 80°C was approximately 40 min.     

A strain of <Emphasis Type="Italic">Meyerozyma guilliermondii</Emphasis> isolated from sugarcane juice is able to grow and ferment pentoses in synthetic and bagasse hydrolysate media

The search for new microbial strains that are able to withstand inhibitors released from hemicellulosic hydrolysis and are also still able to convert sugars in ethanol/xylitol is highly desirable. A yeast strain isolated from sugarcane juice and identified as Meyerozyma guilliermondii was evaluated for the ability to grow and ferment pentoses in synthetic media and in sugarcane bagasse hydrolysate. The yeast grew in xylose, arabinose and glucose at the same rate at an initial medium pH of 5.5. At pH 4.5, the yeast grew more slowly in arabinose. There was no sugar exhaustion within 60 h. At higher xylose concentrations with a higher initial cell concentration, sugar was exhausted within 96 h at pH 4.5. An increase of 350 % in biomass was obtained in detoxified hydrolysates, whereas supplementation with 3 g/L yeast extract increased biomass production by approximately 40 %. Ethanol and xylitol were produced more significantly in supplemented hydrolysates regardless of detoxification. Xylose consumption was enhanced in supplemented hydrolysates and arabinose was consumed only when xylose and glucose were no longer available. Supplementation had a greater impact on ethanol yield and productivity than detoxification; however, the product yields obtained in the present study are still much lower when compared to other yeast species in bagasse hydrolysate. By the other hand, the fermentation of both xylose and arabinose and capability of withstanding inhibitors are important characteristics of the strain assayed.     

局灶性脑缺血再灌损伤时神经细胞内Ca~(2+)时空动态变化的实验研究

本文用插线法制作局灶性脑缺血／再灌损伤模型,利用激光共聚焦扫描显微镜观察活体脑片细胞内Ｃａ２＋的分布及动态变化,结果表明：（１）缺血／再灌时间不同,梗塞面积不同,缺血４小时梗塞面积占同侧半球的１６．３％,缺血４小时再灌２０小时梗塞面积增加到２５．９％,缺血２４小时梗塞面积占同侧半球的６０．４％。（２）本文首次观察到在缺血４小时纹状体区域的Ｃａ２＋变化明显高于皮层,并且再灌后皮层及纹状体区域Ｃａ２＋的含量明显增加     

Effects of resuspension and eutrophication level on summer phytoplankton dynamics in two hypertrophic areas of Lake Taihu,China

The composition of summer phytoplankton communities in two highly eutrophic areas of Lake Taihu, a large, shallow water body in China (area 2,338 km²; mean depth 2.0 m), was studied in relation to selected environmental variables. Both areas have multiple uses, including drinking water supply, fisheries, and recreation. The first area, Meiliang Bay, is much closer to central lake and subjected to wind-driven sediment resuspension, while the second, at Wuli Lake, is less affected by wave action but is considered more nutrient-rich due to its proximity to human inhabitation. By comparing two lake areas, we attempt to advance the understanding and management of different regions of shallow lakes, mainly differing in resuspension levels and eutrophication levels. A comparison of these two lake areas indicated a higher standing crop of phytoplankton in Wuli Lake than in Meiliang Bay. Canonic Correspondence Analysis indicated that nutrient status and sediment resuspension were the principal factors influencing phytoplankton composition in both areas. The dominant algal group in Meiliang Bay comprised immobile R-type strategists (Reynolds in Growth and reproductive strategies of freshwater phytoplankton, 1988), including filamentous diatoms and green algae (Aulacoseira granulate and Planctonema lauterbornii), and recruitment-beneficial RS-type strategists, mainly the colonial cyanobacteria (Microcystis spp.). The community of Wuli Lake was dominated by S-strategists, principally motile flagellates (Cryptomonas spp.) and C-strategists including small nonmotile centric diatoms and chlorococcal green algae.     

Microbial production of 2,3-butanediol from Jerusalem artichoke tubers by <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis>

2,3-Butanediol is one of the promising bulk chemicals with wide applications. Its fermentative production has attracted great interest due to the high end concentration. However, large-scale production of 2,3-butanediol requires low-cost substrate and efficient fermentation process. In the present study, 2,3-butanediol production by Klebsiella pneumoniae from Jerusalem artichoke tubers was successfully performed, and various technologies, including separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF), were investigated. The concentration of target products reached 81.59 and 91.63 g/l, respectively after 40 h in batch and fed-batch SSF processes. Comparing with fed-batch SHF, the fed-batch SSF provided 30.3% higher concentration and 83.2% higher productivity of target products. The results showed that Jerusalem artichoke tuber is a favorable substrate for 2,3-butanediol production, and the application of fed-batch SSF for its conversion can result in a more cost-effective process.     

ROS-Mediated ABA Signaling

In plants, reactive oxygen species (ROS) are short-lived molecules produced through various cellular mechanisms in response to biotic and abiotic stimuli. ROS function as second messengers for hormone signaling, development, oxygen deprivation, programmed cell death, and plant–pathogen interactions. Recent research on ROS-mediated responses has produced stimulating findings such as the specific sources of ROS production, molecular elements that work in ROS-mediated signaling and homeostasis, and a ROS-regulated gene network (Neill et al., Curr Opin Plant Biol 5:388–395, 2002a; Apel and Hirt, Annu Rev Plant Biol 55:373–399, 2004; Mittler et al., Trends Plant Sci 9:490–498, 2004; Mori and Schroeder, Plant Physiol 135:702–708, 2004; Kwak et al., Plant Physiol 141:323–329, 2006; Torres et al., Plant Physiol 141:373–378, 2006; Miller et al., Physiol Plant 133:481–489, 2008). In this review, we highlight new discoveries in ROS-mediated abscisic acid (ABA) signaling. Drs. Daeshik Cho and June M. Kwak are the corresponding authors for this paper.     

TSNAdb: A Database for Tumor-specific Neoantigens from Immunogenomics Data Analysis

Tumor-specific neoantigens have attracted much attention since they can be used as biomarkers to predict therapeutic effects of immune checkpoint blockade therapy and as potential targets for cancer immunotherapy. In this study, we developed a comprehensive tumor-specific neoantigen database (TSNAdb v1.0), based on pan-cancer immunogenomic analyses of somatic mutation data and human leukocyte antigen (HLA) allele information for 16 tumor types with 7748 tumor samples from The Cancer Genome Atlas (TCGA) and The Cancer Immunome Atlas (TCIA). We predicted binding affinities between mutant/wild-type peptides and HLA class I molecules by NetMHCpan v2.8/v4.0, and presented detailed information of 3,707,562/1,146,961 potential neoantigens generated by somatic mutations of all tumor samples. Moreover, we employed recurrent mutations in combination with highly frequent HLA alleles to predict potential shared neoantigens across tumor patients, which would facilitate the discovery of putative targets for neoantigen-based cancer immunotherapy. TSNAdb is freely available at http://biopharm.zju.edu.cn/tsnadb.