A method for isolation of sequences missing in one of two related genomes

We describe a novel technique for isolation of sequences that are present in one genome (tracer), but absent in another (driver). Tracer DNA, cleaved with Sau 3A and capped with a single stranded PCR adapter, is allowed to hybridize with an excess of sheared biotinylated driver; biotinylated DNA and its hybrids with the tracer are removed by phenol/chloroform extraction after incubation with streptavidin. After several rounds of subtraction the ends of self-annealed tracer molecules from the nonextractable fraction are filled-in with Tag polymerase and amplified, using the single stranded PCR adapter as a primer. The method has been applied to purification of fragments from a 2.9 kb plasmid added to E. coli DNA at equimolar quantity. Plasmid derived fragments (250-1000 bp), initially comprising 1/1400th part of tracer DNA, were purified to homogeneity after two rounds of subtraction followed by PCR.     

高频磁场作用下软接触电磁连铸初生坯壳的变形行为

结晶器内初生坯壳的变形行为直接决定着连铸坯的表面质量。笔者运用Reynold润滑方程推导出渣膜内的动压力分布,建立了初生坯壳的二维弹塑性变形有限元模型,研究了施加电磁场前、后坯壳的变形行为。研究结果表明：高频磁场(20kHz)下的感应加热作用会降低保护渣的粘度;电磁压力将增加渣缝的宽度,使施加电磁场后连铸坯初生坯壳的变形行为受到影响,进一步解释了软接触电磁连铸技术改善坯壳表面振痕的机理。     

载体Al/Si比对Cu-ZnO-ZrO2催化剂上CO2加氢合成甲醇的影响

用直接合成法制备了不同铝硅物质的量比Al-SBA-15(β)(β=n(Al)/n(Si)=0,0.02,0.03,0.05)介孔分子筛,并采用浸渍法制备了金属负载量m=35%的CZZ/Al-SBA-15(β)负载型介孔催化剂,考察了Al/Si比对负载催化剂结构和性能的影响。采用N_2吸附-脱附、X射线衍射(XRD)、H_2程序升温还原(H_2-TPR)、CO_2吸附(CO_2-TPD)、NH3吸附(NH3-TPD)和透射电子显微镜(TEM)等手段对样品进行了表征,在固定床反应器上评价了其CO_2加氢合成甲醇的催化性能。实验结果表明,CZZ/Al-SBA-15(β)具有介孔结构,Al的引入不但增加了载体表面酸性位点,同时还提高了催化剂表面铜分散度(DCu%)和铜比表面积(SCu)以及促进了Cu~+-O-Zn活性位的形成,从而大大提高了催化剂的活性。其中CZZ/Al-SBA-15(0.03)催化剂的甲醇选择性达到26%,与不掺铝的CZZ/Al-SBA-15(0)催化剂相比,甲醇选择性提高了11%,但随着掺Al量的继续增加,酸性太强,CO_2吸附能力下降,活性反而降低。     

Caspases cleave focal adhesion kinase during apoptosis to generate a FRNK-like polypeptide

Focal adhesion kinase (Fak) is a non-receptor protein-tyrosine kinase that stimulates cell spreading and motility by promoting the formation of contact sites between the cell and the extracellular matrix (focal adhesions). It suppresses apoptosis by transducing survival signals that emanate from focal adhesions via the clustering of transmembrane integrins by components of the extracellular matrix. We demonstrate that Fak is cleaved by caspases at two distinct sites during apoptosis. The sites were mapped to DQTD772, which was preferentially cleaved by caspase-3, and VSWD704, which was preferentially cleaved by caspase-6 and cytotoxic T lymphocyte-derived granzyme B. The cleavage of Fak during apoptosis separates the tyrosine kinase domain from the focal adhesion targeting (FAT) domain. The carboxyl-terminal fragments that are generated suppress phosphorylation of endogenous Fak and thus resemble a natural variant of Fak, FRNK, that inhibits Fak activity by preventing the localization of Fak to focal adhesions. The cleavage of Fak by caspases may thus play an important role in the execution of the suicide program by disabling the anti-apoptotic function of Fak. Interestingly, rodent Fak lacks an optimal caspase-3 consensus cleavage site although it is cleaved in murine cells undergoing apoptosis at an upstream site. This appears to be the first example of a caspase substrate where the cleavage sites are not conserved between species.     

CBF beta-SMMHC, expressed in M4Eo AML, reduced CBF DNA-binding and inhibited the G1 to S cell cycle transition at the restriction point in myeloid and lymphoid cells

CBF beta-SMMHC is expressed from the inv(16) chromosome in M4Eo AML. Mice lacking CBF subunits or expressing the CBF beta-SMMHC or AML1-ETO oncoproteins failed to develop definitive hematopoiesis. To investigate these effects on hematopoiesis, we expressed CBF beta-SMMHC from the metallothionein promoter, in both 32D cl3 myeloid cells and Ba/F3 B-lymphoid cells. Addition of zinc increased CBF beta-SMMHC levels more than tenfold, with higher levels evident in Ba/F3 lines. Levels obtained in 32D cl3 cells were similar to those of endogenous CBF beta. Indirect immunofluorescence revealed zinc-inducible speckled, nuclear staining in Ba/F3 cells and diffuse nuclear staining in 32D cl3 cells. CBF beta-SMMHC reduced endogenous CBF DNA-binding fivefold in both cell types, increased cell generation time 1.9-fold, on average, in 32D cl3 cells and 1.5-fold in Ba/ F3 cells and decreased tritiated thymidine incorporation into DNA correspondingly. CBF beta-SMMHC increased the proportion of cells in G1 1.7-fold, on average, in 32D cl3 and Ba/F3 cells, and decreased the proportion of cells in S phase by a similar degree. CBF beta-SMMHC induced a marked increase in hypophosphorylated Rb, but did not alter IL-3 Receptor alpha or beta subunit levels. Neither apoptosis nor 32D differentiation was induced by zinc in IL-3 in these lines. Induction of CBF beta-SMMHC in 32D cl3 cells did not inhibit their differentiation to neutrophils or their expression of myeloperoxidase mRNA in G-CSF, and did not produce an eosinophilic phenotype. Additional, proliferative genetic changes in M4eo AMLs might potentiate inhibition of differentiation by CBF beta-SMMHC by allowing its increased expression.     

Chromosomal aberrations, sister-chromatid exchanges, cells with high frequency of SCE, micronuclei and comet assay parameters in 1, 3-butadiene-exposed workers

The association of occupational exposure to 1,3-butadiene (BD) and induction of cytogenetic damage in peripheral lymphocytes was studied in 19 male workers from a monomer production unit and 19 control subjects from a heat production unit. The exposure to BD was measured by passive personal monitors. The following biomarkers were used: chromosomal aberrations (CA), sister chromatid exchanges (SCE), cells with a high frequency of SCE (HFC), micronuclei, comet assay parameters like tail length (TL) and percentage of DNA in tail [T (%)] and polymorphisms of GSTM1 and GSTT1 genotypes. BD exposure with a median value of 0.53 mg/m3 (range: 0.024-23.0) significantly increased (a) the percentage of cells with chromosomal aberrations in exposed vs. control groups (3.11% vs. 2.03%, P<0.01), (b) the frequency of SCE per cell (6.96 vs. 4.87, P<0.001), and (c) the percentage of HFC (19.9% vs. 4.1%, P<0.001). BD exposure had no significant effects on formation of micronuclei and on comet assay parameters. Effect of smoking was observed only for HFC in BD-exposed group. GSTM1 genotype affected chromosomal aberrations in exposed group, while GSTT1 genotype affected chromosomal aberrations in controls. No effect of GSTM1 or GSTT1 genotypes was observed on any other biomarkers used.     

猴头菌子实体与液态发酵菌丝体活性成分差异及其在改善胃部疾病中的应用进展

猴头菌是传统的食药两用菌,具有猴头菌多糖、猴头菌素、猴头菌酮、麦角甾醇等多种活性成分,对胃炎、胃溃疡、胃癌等胃部疾病具有有益调节作用。猴头菌有子实体和菌丝体两种形态,菌丝体又分为固态发酵菌丝体与液态发酵菌丝体。本文主要对比阐述了猴头菌子实体与液态发酵菌丝体活性成分差异以及它们在改善胃部疾病中相关研究成果,以期为猴头菌液态发酵菌丝体规模化、工业化、自动化生产,以及功能食品和食用菌基天然药物的开发提供依据和参考。     

Therapeutic changes in extraction versus non-extraction orthodontic treatment

A family cancer database was constructed from the nationwide Swedish registries and includes approximately 6 million persons and >30,000 cancers in offspring diagnosed at ages 15-51 years and their parents. A particular advantage of the database is that the contribution of both parental lineages on cancer risk can be examined. Cancer risk in the offspring was increased approximately 1.1 times when the father had cancer, and no increase was noted when the mother had cancer. If both parents had cancer, the risk for sons was 1.4 and for daughters 1.3. The sites of increased cancer risk in the offspring were colorectum, breast, cervix, corpus uteri, ovary, testis, melanoma, eye, other endocrine glands, and multiple myeloma. The results among young and middle-age adults suggest that cancer in both parents increases the cancer risk in the offspring at many sites. The molecular genetic explanation may be that rare dominant single genes increase susceptibility at many sites, or that overlapping sets of genes control susceptibility at multiple sites.     

不同工艺状态的玻璃布/酚醛复合材料防隔热特性研究

玻璃布/酚醛复合材料有较好的耐烧蚀和隔热特性,被广泛用于工业耐高温设备以及高速飞行的航天器防隔热设计上,但是不同的成型工艺表现出的耐烧蚀及隔热特性却存在差异。将不同玻璃布厚度、不同缠绕工艺、不同固化方式、以及有机纤维与玻璃纤维混编工艺获得的玻璃布/酚醛制品,取样进行风洞条件下的耐烧蚀、隔热性能试验,试验测量了各模型的质量烧蚀率及背面温升,用于评估其相应的防隔热性能。试验结果显示倾斜缠绕工艺耐烧蚀及隔热性能最优,与理论预期一致,可为工程应用提供参考。