Metabolomic approach to optimizing and evaluating antibiotic treatment in the axenic culture of cyanobacterium Nostoc flagelliforme

The application of antibiotic treatment with assistance of metabolomic approach in axenic isolation of cyanobacterium Nostoc flagelliforme was investigated. Seven antibiotics were tested at 1–100 mg L^?1, and order of tolerance of N. flagelliforme cells was obtained as kanamycin > ampicillin, tetracycline > chloromycetin, gentamicin > spectinomycin > streptomycin. Four antibiotics were selected based on differences in antibiotic sensitivity of N. flagelliforme and associated bacteria, and their effects on N. flagelliforme cells including the changes of metabolic activity with antibiotics and the metabolic recovery after removal were assessed by a metabolomic approach based on gas chromatography–mass spectrometry combined with multivariate analysis. The results showed that antibiotic treatment had affected cell metabolism as antibiotics treated cells were metabolically distinct from control cells, but the metabolic activity would be recovered via eliminating antibiotics and the sequence of metabolic recovery time needed was spectinomycin, gentamicin > ampicillin > kanamycin. The procedures of antibiotic treatment have been accordingly optimized as a consecutive treatment starting with spectinomycin, then gentamicin, ampicillin and lastly kanamycin, and proved to be highly effective in eliminating the bacteria as examined by agar plating method and light microscope examination. Our work presented a strategy to obtain axenic culture of N. flagelliforme and provided a method for evaluating and optimizing cyanobacteria purification process through diagnosing target species cellular state.     

六种鱼腥藻在大量培养中的生产率和适应性与营养成分比较

连续两年对鱼腥藻的六个品系进行了大量培养比较研究,在长江中下游气候条件下,HB1042和HB1105具有稳定的生产性能,5月下旬至九月中旬123d内,平均有效生长分别为114和100d;其次是HB1058,HB686和HB1017,平均有效生长天分别为73,58和71d;HB13适应性较差。生物量生产率在5月下旬、6月中旬至9月中旬较高,除HB13外,平均都在10g干重/m~2/d以上或接近10g/m~2/d。在不同的季节它们表现出了各自的最高生物量生产率。据此,提出了逐月逐旬采取藻种搭配生产,以获得最高生物量生产率的配合关系。营养成分分析表明它们的蛋白质含量在40％左右;氨基酸组成合理,除某些种类的含硫氨基酸略低外,都符合FAO/WHO的标准。     

RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation

In acute promyelocytic leukaemia (APL), the promyelocytic leukaemia (PML) protein is fused to the retinoic acid receptor alpha (RAR). This disease can be treated effectively with arsenic, which induces PML modification by small ubiquitin-like modifiers (SUMO) and proteasomal degradation. Here we demonstrate that the RING-domain-containing ubiquitin E3 ligase, RNF4 (also known as SNURF), targets poly-SUMO-modified proteins for degradation mediated by ubiquitin. RNF4 depletion or proteasome inhibition led to accumulation of mixed, polyubiquitinated, poly-SUMO chains. PML protein accumulated in RNF4-depleted cells and was ubiquitinated by RNF4 in a SUMO-dependent fashion in vitro. In the absence of RNF4, arsenic failed to induce degradation of PML and SUMO-modified PML accumulated in the nucleus. These results demonstrate that poly-SUMO chains can act as discrete signals from mono-SUMOylation, in this case targeting a poly-SUMOylated substrate for ubiquitin-mediated proteolysis.     

含par位点的重组质粒Psjm3的构建及其稳定性研究

利用自然质粒pSC101par位点的分离稳定性功能,构建了含par位点的质粒pSJM4和pSJM3,通过在同样宿主E.coli HB101中的稳定性比较研究表明,不含par位点的重组质粒pSJ3很不稳定,E.coli G3(pSJ3)在培养到第10代时已开始出现pSJ3的丢失,到培养至50代时则已全部丢失;而含par位点的重组质粒pSJM3则表现得十分稳定,E.coli G3-1(pSJM3)经70代培养,仍无明显的质粒丢失现象,其稳定率保持97%以上。通过对不含par和含par的非重组质粒pUC18和pSJM4的稳定性比较也获得同样的结果。通过对E.coliG3(pSJ3)和E.coli G3-1(pSJM3)的产酶活性比较研究表明,G3-1菌株明显高于G3菌株,说明我们构建的重组质粒pSJM3上的par位点功能不仅没有因外源基因的表达而受影响,而且有利于外源基因的表达。     

红壤茶树根层土壤基础呼吸作用和酶活性

对不同树龄茶树根层土壤的呼吸作用(包括代谢熵qCO2)和土壤酶(脲酶、转化酶和酸性磷酸单酯酶)活性进行了研究、不同树龄茶树根层土壤日基础呼吸作用强度(36．23—58．52mg·kg^-1·d^-1)和日代谢墒(0．30一0．68)都以40和90年茶树较为接近,分别显著大于和小于10年树龄茶树根层土壤;服酶活性(41．48—47、72mg·kg^-1·d^-1)则三者间差异不大,虽然随树龄增长而下降;转化酶活性(189．29—363．40mg·kg^-1·d^-1)也随树龄增长而下降,并且10年茶树根层土壤显著大于40和90年树龄茶树;而酸性磷酸单酯酶活性(444．22—828．32mg·kg^-1·d^-1)相反,随树龄增长而增强．结果表明,土壤基础呼吸作用、代谢熵和3种土壤酶活性都与茶树树龄、土壤pH、土壤有机碳、土壤全氮、土壤可活性酚总量、及土壤微生物生物量密切相关．     

Tissue microarray study for classification of breast tumors

Clinical and pathological heterogeneity of breast cancer hinders selection of appropriate treatment for individual cases. Molecular profiling at gene or protein levels may elucidate the biological variance of tumors and provide a new classification system that correlates better with biological, clinical and prognostic parameters. We studied the immunohistochemical profile of a panel of seven important biomarkers using tumor tissue arrays. The tumor samples were then classified with a monothetic (binary variables) clustering algorithm. Two distinct groups of tumors are characterized by the estrogen receptor (ER) status and tumor grade (p = 0.0026). Four biomarkers, c-erbB2, Cox-2, p53 and VEGF, were significantly overexpressed in tumors with the ER-negative (ER-) phenotype. Eight subsets of tumors were further identified according to the expression status of VEGF, c-erbB2 and p53. The malignant potential of the ER-/VEGF+ subgroup was associated with the strong correlations of Cox-2 and c-erbB2 with VEGF. Our results indicate that this molecular classification system, based on the statistical analysis of immunohistochemical profiling, is a useful approach for tumor grouping. Some of these subgroups have a relative genetic homogeneity that may allow further study of specific genetically-controlled metabolic pathways. This approach may hold great promise in rationalizing the application of different therapeutic strategies for different subgroups of breast tumors.