Microdiversity of uncultured marine prokaryotes: the SAR11 cluster and the marine Archaea of Group I

The SAR11 cluster and the Group I of marine Archaea represent probably the best two examples of uncultured marine prokaryotes of widespread occurrence. To study their microdiversity and distribution, a total of 81 and 48 clones, respectively, were sequenced from Mediterranean and Antarctic waters at different locations and depths. The DNA regions chosen for the analysis were the last third, approximately, of the 16S rRNA gene and the 16S-23S intergenic spacer (also known as internal transcribed spacer [ITS]). There was a high concordance in both, even with the extremely variable ITS, where potential probes have been proposed for the identification and isolation of these micro-organisms. In terms of community structure, our results show that although depth-related factors seem to be predominant in the final associations of the clones, geography also plays a significant role. A major group of surface-associated sequences was found in both SAR11 and marine Archaea. In both cases this group was relatively homogeneous containing little diversity in terms of sequence, while sequences retrieved from deep samples and some surface clones contained much more heterogeneity. As a whole, both groups of prokaryotes seem to fall within the limits of well-defined taxonomic units.     

Calcifying epithelial odontogenic tumor: an immunohistochemical case study

Calcifying epithelial odontogenic tumor (CEOT) is a rare benign odontogenic tumor. A case of CEOT in a 25-year-old female is presented here. Histologically, the case showed sheets of polyhedral epithelial cells with deep eosinophilic cytoplasm and prominent nuclei. Nuclear pleomorphism and hyperchromatism were evident. Globules of amyloid-like material among the tumor cells were prominent. Also found was a small area demonstrating a cribriform pattern. Immunohistochemical studies with antibodies against basement membrane proteins (laminins 1 and 5, collagen type IV and fibronectin), pan-cytokeratins AE1/AE3, vimentin, S-100 protein and CD 1a were performed. Tumor cells expressed laminins 1 and 5, fibronectin, cytokeratins and vimentin. The amyloid-like material was not reactive to all antibodies examined. A number of dendritic cells among sheets of tumor cells were revealed with strong staining for S-100 protein and CD 1a. These dendritic cells are likely to be Langerhans cells. Hence, immunohistochemistry is a useful method to study the variant of CEOT.     

Construction of Lactobacillus plantarum strain with enhanced L-lysine yield

AIM: To enhance L-lysine secretion in Lactobacillus plantarum. METHODS AND RESULTS: An S-2-aminoethyl-L-cystein (AEC)-resistant mutant of L. plantarum was isolated, and it produced L-lysine at considerably higher level than the parent strain. Aspartokinase in the mutant has been desensitized to feedback inhibition by L-lysine. The nucleotide sequence analysis of thrA2 that codes for aspartokinase in the mutant predicted a substitution of glutamine to histidine at position 421. L-Lysine-insensitive aspartokinase, together with aspartate semialdehyde dehydrogenase, dihydrodipicolinate synthase, and dihydrodipicolinate reductase genes, was cloned from L. plantarum DNA to a shuttle vector, pRN14, and the genes were then transformed individually into the AEC-resistant mutant and the parent strain. The overexpression of the genes led to the increase in the activity of enzymes they encode in vitro. However, only the strain overexpressing aspartokinase or dihydrodipicolinate synthase produced more L-lysine. CONCLUSIONS: The desensitization of aspartokinase to L-lysine in L. plantarum led to the overproduction of L-lysine. The overexpression of L-lysine-insensitive aspartokinase or dihydrodipicolinate synthase enhanced L-lysine secretion in L. plantarum. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of the L-lysine-overproducing strain of L. plantarum in food or feed fermentation may increase the L-lysine content of fermented products.     

P-gp、TopoⅡ在非小细胞肺癌中的表达

为明确化疗前非小细胞肺癌（nonsmall cell lung cancer,NSCLCL)中P－糖蛋白（P－Glycoprotein,P-gp)、拓扑异构酶Ⅱ（TopoisomeraseⅡ,TopoⅡ）的不同表达与NSCLC的组织类型、细胞分化程度、TNM分期是否相关,是否存在原发耐药,是否影响NSCLC患的预后,本实验采用S－P免疫组化方法,检测97例化疗前NSCLC患中P-gp、TopoⅡ的表达情况。结果表明：P－gp在腺癌中高表达,在有淋巴结转移的病例中高表达,且P-gp和TopoⅡ在表达上虽不具有相关性,但两表达程度及其交互作用都是NSCLC患预后的影响因素。     

蕲蛇酶抗小鼠实验性肿瘤转移作用研究

目的：探讨从尖吻蝮蛇毒中分离得到的具有凝血酶样酶活性的蕲蛇酶抗实验性肿瘤转移作用。方法：用尾静脉注射体外培养的黑色素瘤Ｂ１６和肉瘤Ｓ－１８０细胞的小鼠肺转移模型,注射瘤细胞前后分别腹腔注射药物,２０天后处死小鼠,计数肺表面转移瘤结节数。结果蕲蛇酶剂量在２－５ＡＵ／ｋｇｉｐ能明显减少Ｂ１６在Ｃ５７ＢＬ小鼠及Ｓ－１８０在昆明鼠的肺转移结节数,但对转移瘤小鼠的生命无明显的延长作用。结论蕲蛇酶具有抗小鼠实     

一株具霉菌抑制活性细菌的鉴定

为探讨菌株HB02对霉菌生长的影响,将HB02与霉菌孢子悬液同时加入到PYG肉汤,在28℃培养15d。在培养的第3、69、、12和15天测定培养液中的黄曲霉和禾谷镰刀菌菌丝重量。结果显示:与对照组相比,HB02可显著抑制黄曲霉和禾谷镰刀菌生长(P<0.01)。通过常规细菌学实验鉴定该菌株为一株乳酸杆菌。通过分子生物学方法测定了菌株的16S rRNA基因序列,进一步证实了HB02为一株乳酸杆菌。根据Berthier等的方法,通过HB0216S/23S rRNA基因间隔(Intergenic Spacer Regions,ISR)序列的扩增和测定,再通过基因文库中所有细菌基因序列比较分析,最终鉴定菌株HB02为一株弯曲乳酸杆菌(Lactobacillus curvatus)。     

中间偃麦草(Thinopyrum intermedium)18S-5.8S-26S rDNA位点在染色体上的分布及对其核ITS序列异质性的分析(英文)

中间偃麦草(Thinopyrum intermedium(Host)Barkworth et Dewey)是禾本科小麦族植物中的一个异源六倍体物种,是重要的牧草植物,在小麦的抗病育种中发挥了重要作用。利用荧光原位杂交(FISH)技术,在体细胞中期染色体上,对18S-5.8S-26S rDNA位点进行了物理定位,发现该物种有3～4对染色体携带18S-5.8S-26S rDNA主位点。结合基因组原位杂交(GISH)分析,证明中间偃麦草的St基因组中有一对同源染色体短臂末端携带一个主位点,其余2～3对主位点位于E基因组染色体上。对不同来源的材料研究表明:18S-5.8S-26S rDNA位点的数目(包括主位点和小位点)、位置、拷贝数在不同收集材料之间的差异较大,甚至在同一个体的不同细胞中也存在差异。讨论了rDNA物理作图数据在分析系统发育问题中的局限性。结合中间偃麦草的三个可能的二倍体基因组供体(Th.bessarabicum、Th. elongatum和Pseudoroegneria stipifolia)rDNA位点分析的结果,对中间偃麦草进化过程中rDNA位点的变化进行了分析,同时,对其中一份材料的核ITS序列进行了克隆、测序和系统发育分析,发现在中间偃麦草中,ITS序列具有很高的异质性。     

烯效唑干拌种对小麦叶片衰老期间有关酶活性的影响

研究不同浓度(0、10、20、40mg·kg-1)烯效唑干拌种对小麦品种川麦30不同叶序(3叶、7叶、旗叶)叶片衰老期间酶活性影响的结果表明,烯效唑干拌种后,不同叶序叶片超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性增强,衰老后期仍维持较高水平;而核糖核酸酶(RNase)活性水平及上升速率则受抑,叶片中丙二醛(MDA)积累量减少,可溶性蛋白质含量下降缓慢.     

面包酵母催化羰基不对称还原合成手性醇的研究

以2-辛酮和4-氯乙酰乙酸乙酯(COBE)为模型底物分别考察了酵母细胞对直链甲基酮和陆羰基酯中的羰基不对称还原情况。实验发现不对称还原2-辛酮的产物主要是S型的2-辛醇,且对映体选择性很高。不对称还原COBE生成的主要是S(D)-型产物,反应COBE的转化率、光学选择性都比较高。同时发现COBE的浓度和产物对不对称还原都有一定负面的影响。     

IKK regulates the deubiquitinase CYLD at the postsynaptic density

K63-linked polyubiquitination of proteins regulates their trafficking into specific cellular pathways such as endocytosis and autophagy. CYLD, a deubiquitinase specific for K63-linked polyubiquitins, is present in high quantities at the postsynaptic density (PSD). It was previously shown that, under excitatory conditions, CaMKII activates CYLD in a Ca²⁺-dependent manner. The observation that CYLD can also be phosphorylated in the absence of Ca²⁺ in isolated PSDs led us to further explore the regulation of CYLD under basal conditions. A possible involvement of the autonomous form of CaMKII and IKK, both kinases known to be localized at the PSD, was examined. A CaMKII inhibitor CN21 had no effect on CYLD phosphorylation in the absence of Ca²⁺, but two different IKK inhibitors, IKK16 and tatNEMO, inhibited its phosphorylation. Immuno-electron microscopy on hippocampal cultures, using an antibody for CYLD phosphorylated at S-418, revealed that the phosphorylated form of CYLD is present at the PSD under basal conditions. Phosphorylation of CYLD under basal conditions was inhibited by IKK16. NMDA treatment further promoted phosphorylation of CYLD at the PSD, but IKK16 failed to block the NMDA-induced effect. In vitro experiments using purified proteins demonstrated direct phosphorylation and activation of CYLD by the beta catalytic subunit of IKK. Activation of IKK in isolated PSDs also promoted phosphorylation of CYLD and an increase in endogenous deubiquitinase activity for K63-linked polyubiquitins. Altogether, the results suggest that in the absence of excitatory conditions, constitutive IKK activity at the PSD regulates CYLD and maintains basal levels of K63-linkage specific deubiquitination at the synapse.