Synthetic statistical approach reveals a high degree of richness of microbial eukaryotes in an anoxic water column

Molecular surveys suggest that communities of microbial eukaryotes are remarkably rich, because even large clone libraries seem to capture only a minority of species. This provides a qualitative picture of protistan richness but does not measure its real extent either locally or globally. Statistical analysis can estimate a community's richness, but the specific methods used to date are not always well grounded in statistical theory. Here we study a large protistan molecular survey from an anoxic water column in the Cariaco Basin (Caribbean Sea). We group individual 18S rRNA gene sequences into operational taxonomic units (OTUs) using different cutoff values for sequence similarity (99 to 50%) and systematically apply parametric models and nonparametric estimators to the OTU frequency data to estimate the total protistan diversity. The parametric models provided statistically sound estimates of protistan richness, with biologically meaningful standard errors, maximal data usage, and extensive model diagnostics and were preferable to the available nonparametric tools. Our clone library exceeded 700 clones but still covered only a minority of species and less than half of the larger protistan clades. Our estimates of total protistan richness portray the target community as very rich at all OTU levels, with hundreds of different populations apparently co-occurring in the small (3-liter) volume of our sample, as well as dozens of clades of the highest taxonomic order. These estimates are among the first for microbial eukaryotes that are obtained using state-of-the-art statistical methods and can serve as benchmark numbers for the local diversity of protists.     

干旱区多枝柽柳的生长特性

多枝柽柳是分布最为广泛的柽柳属植物之一。也是维持干旱区荒漠生态系统的关键植物种。本研究运用树轮年代学和树干解析方法。对额济纳地区不同生境条件下多枝柽柳的年轮生长、高生长和地径生长等方面的特征进行初步探讨。多枝柽柳每年形成1个年轮。树盘偏心生长极为普遍．完整树轮大多在10龄以内区域,10龄以外都呈单个或多个不完整年轮生长区。多枝柽柳单枝在1～4年高生长缓慢．4～8龄之间属于快速生长期,10年以上生长速率急剧下降。不同地下水位条件下三组样本径生长年增长速率达到极大值的年龄分别为21、56和74年,水环境条件是影响多枝柽柳径向生长的主要因素。同一年年轮宽度随枝高呈减小趋势。多枝柽柳边材的树轮数一般在3～5轮之间,进入老龄阶段有增加趋势。边材宽度随年龄增长逐渐减小。     

牛血清白蛋白对小鼠原核期胚胎玻璃化冷冻的影响

以小鼠原核期胚胎为对象,以胚胎的存活率、卵裂率、囊胚率以及囊胚细胞数作为检测指标,在M2液的基础上添加8种浓度(0,2,4,8,16,32,64,96mg/mL)牛血清白蛋白(BSA)配置防冻液,探讨防冻液和玻璃化冷冻后对胚胎发育的影响。BSA防冻液对胚胎发育影响的实验结果表明,8个浓度组间以及与对照组间胚胎的卵裂率、囊胚率以及囊胚细胞数无显著差异(P>0.05),说明在防冻液中加入一定浓度的BSA对小鼠胚胎无毒性作用。防冻液经玻璃化冷冻后对胚胎发育影响的实验表明,8个浓度组间冷冻胚胎复苏后的存活率、卵裂率、囊胚率及囊胚细胞数无显著差异(P>0.05)。表明BSA在这种防冻液中没有明显的保护作用。从经济、实用、生物安全角度考虑,不支持在玻璃化防冻液中添加BSA。     

不同诱捕技术对松褐天牛的诱捕效果

通过不同引诱剂、不同诱芯、不同诱捕器和不同缓释量对松褐天牛的诱捕试验,结果表明,4种引诱剂对松褐天牛都有一定的引诱效果.其中,引诱剂MA2K 05的效果最好,引诱活性平均为26.3头/诱捕器,对其它鞘翅目和半翅目昆虫也有引诱作用;引诱剂MA2K 13活性次之,平均21.3头/诱捕器;MA2K 11最弱,平均13.8头/诱捕器.3种诱芯A、B和C的引诱效果分别为14.73、4.25和20.3头/诱捕器,其中诱芯C内引诱剂总量为诱芯A或诱芯B的2.5倍,且诱芯C内的引诱剂3～5 d需添加一次,而诱芯A和诱芯B的引诱活性能持续1个月以上,因此,诱芯B较为理想.宣州诱捕器引诱效果显著高于日式诱捕器,分别为36.4和9.7头/诱捕器.不同剂量诱捕结果表明,随着诱芯缓释量的增加,诱捕效果也相应提高,当剂量从20 ml上升至80 ml时,诱捕效果增加不显著;当诱剂达120 ml时,引诱效果显著增加.     

松褐天牛触角感器电镜扫描和触角电位反应

应用扫描电镜对松褐天牛的触角感器进行扫描观察,并测定了触角对松树主要挥发物的电生理反应.松褐天牛触角上共有7种感器,即毛形感器、锥形感器、指形感器、棒形感器、瓶形感器、芽形感器以及刺形感器,以毛形和锥形感器的数量最丰富.其中毛形感器和锥形感器各有3个型,指形感器有2个型.毛Ⅱ、毛Ⅲ型和锥Ⅰ、锥Ⅱ型感器表皮结构具有深的纵刻纹,具有嗅觉感器的典型特征.同时对各种感器的数量、长度及分布特点等作了描述.电生理研究结果表明,几种供试化合物及其混合物均能引起松褐天牛雌、雄成虫较强的电生理反应;松褐天牛触角不同部位的EAG反应存在显著差异;剂量试验表明,松褐天牛成虫触角对不同浓度的α-Pinene的EAG反应存在一定的规律性.     

E.coli ZB43生成Microcin B17的发酵研究

Microcin B17(简称MccB17)是核糖体合成的肽类DNA螺旋酶抑制剂,其独特的分子结构是抗菌剂未来发展的重要的新颖结构,MccB17产生菌的杂环形成机理对于制药工业的药物设计具有重要意义。MccB17产生菌E.coliZB43适宜在M63合成培养基中积累胞内MccB17,低溶氧水平利于MccB17的积累。当葡萄糖浓度超过1g/L时,MccB17的合成受到阻遏。采用均匀设计表U11(1110)进行发酵培养基配比调整,确定了最佳的含葡萄糖合成培养基配比:葡萄糖1g/L,KH2PO43g/L,K2HPO47g/L,(NH4)2SO41g/L,MgSO4.7H2O 1.8mmol/L,盐酸硫胺1.8μg/ml。丁二酸钠能解除葡萄糖对E.coliZB43生成MccB17的代谢阻遏。以10g/L丁二酸钠替代葡萄糖为唯一碳源,在上述最佳发酵培养基配比条件下,37℃24h,MccB17的产量可达559.6μg/ml。     

Survivin protein expression in bovine follicular oocytes and their in vitro developmental competence

This study examined the relationship between survivin expression and the stage of development of in vitro cultured bovine oocytes and embryos; and whether survivin expression is affected by the quality of cumulus–oocyte complexes (COCS) or the quality of pre-implantation embryos. A polyclonal antibody was prepared using recombinant bovine survivin protein. Expression of survivin mRNA and protein was analyzed by real-time quantitative RT-PCR and immunocytochemistry. In the first experiment, survivin mRNA expression was examined at developmental stages from germinal vesicle (GV) oocyte to blastocyst, it was significantly decreased after fertilization of matured oocytes (P < 0.05), then increased slightly to the 8-cell stage followed by rapid increases at the morula and blastocyst stages (P < 0.05). In the second experiment, the effect of oocyte quality on survivin protein, pro-apoptotic (bax, caspase-3) and anti-apoptotic (survivin, bax inhibitor) mRNA expression was examined. Survivin protein was more strongly expressed in good quality COCS than in poor quality COCS. The expression of the anti-apoptotic genes, survivin and bax inhibitor, was significantly higher (P < 0.05) and that of the pro-apoptotic genes, bax and caspase-3, was significantly lower (P < 0.05) in good compared to poor quality COCS. The developmental competence of good quality COCS (30.4% blastocysts) was significantly better than that of poor quality COCS. In the last experiment also, we confirmed that significantly higher expression of survivin and bax inhibitor genes and significantly lower expression of bax and caspase-3 genes was resulted in good quality blastocysts than in poor quality blastocysts (P < 0.05). It was concluded that the expression of survivin was related to the quality of COCS, their developmental competence and the quality of in vitro produced blastocysts. Consequently, survivin may be a good candidate marker for embryo quality.     

Genetic characteristics of 207 microsatellite markers in the Korean population and in other Asian populations

Microsatellites, short tandem repeats, are useful markers for genetic analysis because of their high frequency of occurrence over the genome, high information content due to variable repeat lengths, and ease of typing. To establish a panel of microsatellite markers useful for genetic studies of the Korean population, the allele frequencies and heterozygosities of 207 microsatellite markers in 119 unrelated Korean, Indian and Pakistani individuals were compared. The average heterozygosity of the Korean population was 0.71, similar to that of the Indian and Pakistani populations. More than 80% of the markers showed heterozygosity of over 0.6 and were valuable as genetic markers for genome-wide screening for disease susceptibility loci in these populations. To identify the allelic distributions of the multilocus genetic data from these microsatellite markers, the population structures were assessed by clustering. These markers supported, with the most probability, three clustering groups corresponding to the three geographical populations. When we assumed only two hypothetical clusters (K), the Korean population was separate from the others, suggesting a relatively deep divergence of the Korean population. The present 207 microsatellite markers appear to reflect the historical and geographical origins of the different populations as well as displaying a similar degree of variation to that seen in previously published genetic data. Thus, these markers will be useful as a reference for human genetic studies on Asians.     

Preparation of sodium deoxycholate (DOC) conjugated heparin derivatives for inhibition of angiogenesis and cancer cell growth

We describe new DOC (sodium deoxycholate)-heparin nanoparticles for in vivo tumor targeting and inhibition of angiogenesis based on chemical conjugation and the enhanced permeability and retention (EPR) effect. Heparin has been used as a potent anticoagulant agent for 70 years, and has recently been found to inhibit the activity of growth factors which stimulate the smooth muscle cells around tumor. From the results, DOC and heparin were conjugated by bonding carboxyl groups of heparin with amine groups of aminated sodium deoxycholate. Larger antitumor effects of the DOC-heparin VI (8.5 mol of DOC coupled with 1.0 mol heparin) were achieved in animal studies, compared to heparin alone. We confirmed that the conjugated heparin retained its ability to inhibit binding with angiogenic factor, showing a significant decrease in endothelial tubular formation. These results provide new insights into the nontoxic anticancer drug carrier as well as the design of multifunctional bioconjugates for targeted drug delivery.