Fifteen novel polymorphic microsatellites in rock carp, Procypris rabaudi (Tchang), an endemic fish species in the upper reaches of the Yangtze River drainage

Fifteen new microsatellite markers were isolated in rock carp, Procypris rabaudi (Tchang), from an AC-enriched genomic library. The allele number of these loci ranged from three to 13 (average 6.87 per locus) in the tested 24 individuals. Polymorphism information content (PIC) ranged from 0.453 to 0.821 with an average of 0.719. Average observed and expected heterozygosities were 0.712 (range 0.565–0.917) and 0.772 (range 0.542–0.855), respectively. These microsatellite markers could be useful in the population genetic study of rock carp.     

The institute for genomic research Osa1 rice genome annotation database

We have developed a rice (Oryza sativa) genome annotation database (Osa1) that provides structural and functional annotation for this emerging model species. Using the sequence of O. sativa subsp. japonica cv Nipponbare from the International Rice Genome Sequencing Project, pseudomolecules, or virtual contigs, of the 12 rice chromosomes were constructed. Our most recent release, version 3, represents our third build of the pseudomolecules and is composed of 98% finished sequence. Genes were identified using a series of computational methods developed for Arabidopsis (Arabidopsis thaliana) that were modified for use with the rice genome. In release 3 of our annotation, we identified 57,915 genes, of which 14,196 are related to transposable elements. Of these 43,719 non-transposable element-related genes, 18,545 (42.4%) were annotated with a putative function, 5,777 (13.2%) were annotated as encoding an expressed protein with no known function, and the remaining 19,397 (44.4%) were annotated as encoding a hypothetical protein. Multiple splice forms (5,873) were detected for 2,538 genes, resulting in a total of 61,250 gene models in the rice genome. We incorporated experimental evidence into 18,252 gene models to improve the quality of the structural annotation. A series of functional data types has been annotated for the rice genome that includes alignment with genetic markers, assignment of gene ontologies, identification of flanking sequence tags, alignment with homologs from related species, and syntenic mapping with other cereal species. All structural and functional annotation data are available through interactive search and display windows as well as through download of flat files. To integrate the data with other genome projects, the annotation data are available through a Distributed Annotation System and a Genome Browser. All data can be obtained through the project Web pages at http://rice.tigr.org.     

Identification of an essential cleavage site in ColE7 required for import and killing of cells

Colicin E7 (ColE7), a nuclease toxin released from Escherichia coli, kills susceptible bacteria under environmental stress. Nuclease colicins are processed during translocation with only the cytotoxic nuclease domains traversing the inner membrane to cleave tRNA, rRNA, or DNA in the cytoplasm of target cells. In this study, we show that the E. coli periplasmic extract cleaves ColE7 between Lys(446) and Arg(447) in the presence or absence of its inhibitor Im7 protein. Several residues near cleavage sites were mutated, but only mutants of Arg(447) completely lost in vivo cell-killing activity. Both the full-length and the nuclease domain of Arg(447) mutants retained their nuclease activities, indicating that failure to kill cells was not a consequence of damage to the endonuclease activity of the enzyme. Moreover, the R447E ColE7 mutant was not cleaved at its 447 site by periplasmic extracts or transported into the cytoplasm of target cells. Collectively, these results suggest that ColE7 is cleaved at Arg(447) during translocation and that cleavage is an essential step for ColE7 import into the cytoplasm of target cells and its cell-killing activity. Conserved basic residues aligned with Arg(447) have also been found in other nuclease colicins, implying that the processing at this position may be common to other colicins during translocation.     

Schistosoma japonicum: a method for transformation by electroporation

Despite of our knowledge of genetic make up of schistosomes, a number of genes have not been characterized largely due to lack of effective transformation protocols. Here we present electroporation as a strategy for effective introduction of plasmids DNA into schistosomula and adults. Using plasmids of pEGFP-C1 as an expression vector, we first verified that the CMV promoter could direct EGFP to express in primary culture cells from Schistosoma japonicum. Subsequently, the plasmids were introduced into schistosomula and adults by electroporation and EGFP expression was demonstrated using molecular and microscopical methods. Our findings indicate that electroporation is an effective method for transformation of S. japonicum.     

A continuous fluorescent assay for measuring protease activity using natural protein substrate.

A continuous caseinolytic activity assay has been developed and characterized with trypsin, a serine protease, and transin, a metalloproteinase. Beta-casein labeled with both N-(7-dimethylamino-4-methylcoumarinyl)-maleimide (DACM) and fluorescein isothiocyanate (FITC) is used as the substrate in this assay. The effect of proteolysis of the substrate is a reduction of the intermolecular energy transfer from DACM to FITC. The caseinolytic activity is then monitored by the fluorescence increase. The activities of both proteases obey Michaelis-Menten kinetics with Km = 1.6 +/- 0.2 microM for trypsin and Km = 13.2 +/- 1.9 microM for transin. Protease concentrations as low as 10 ng/mL can be utilized. The pH dependence of the caseinolytic activity has been determined for both enzymes.     

Evidence of compliance with and effectiveness of guidelines for noninvasive prenatal testing in China: a retrospective study of 189,809 cases

In China,the medical guidelines recommend performing noninvasive prenatal testing (NIPT) with caution for pregnant women aged 35 years or older.However,the Mother and Child Health Care Law suggests that all primiparous women whose age is older than 35 years undergo prenatal diagnosis.These two inconsistent suggestions/recommendations have made obstetricians confused about whether to offer NIPT to these older pregnant women.To face this issue and find out the solution we performed a retrospective study of 189,809 NIPT samples collected from 28 provincial-leveled administrative units in China.Of 1,564women with high-risk pregnancies who underwent NIPT,459 (29.3%) did not participate in follow-up.The compound sensitivity and specificity of NIPT for trisomies 21,18 and 13 detection was 99.1%(95%CI,98.0%-99.6%) and 99.9%(95%CI,98.8%-99.9%),respectively.In secundiparous women,NIPT showed high sensitivity and specificity similar to that in primiparous women.The observed risk for trisomies 21 and 18 significantly increased when the maternal age was 39 and older.After the publication of the current NIPT policy,the follow-up rate at our center was 97.9%;however,a large number of women are not in maternal and infant care networks nationwide,and that makes the follow-up rate outside our center relatively low.Our study shows that to balance the prevention of major aneuploidies and the limited resources for prenatal diagnosis,the cut-off age of 35for invasive prenatal diagnosis might be unnecessary.Although the NIPT guidelines are well written,how to practice it effectively,especially in less industrialized areas,is worth discussing.     

高粱种质材料幼苗期耐盐碱性评价

采用Hoagland营养液砂培法,以NaCl和Na2CO3组成的混合盐碱对高粱幼苗进行胁迫处理,建立高粱幼苗期耐盐碱评价方法,并评价了66份高粱种质材料的耐盐碱性.结果表明:盐浓度在8.0～12.5 g·L-1时,高粱耐盐碱品种‘TS-185’与盐碱敏感品种‘Tx-622B’在幼苗期的耐盐碱性差异明显,表明进行高粱幼苗期耐盐碱性评价时适宜的盐浓度范围为8.0～12.5 g· L-1.在10.0和12.5 g·L-12个盐浓度下,66份高粱种质材料的相对存活率、相对地上部鲜质量和相对株高的差异均达显著水平,表明不同品种的耐盐碱性不同.其中,‘三尺三’为高度耐盐碱品种,‘MN-2735’等16个品种为耐盐碱品种,‘EARLY HONEY’等32个品种为中等耐盐碱品种,‘Tx-622B’等16个品种为盐碱敏感品种,‘MN-4588’为高度盐碱敏感品种.苏丹草类型高粱一般具有较高的耐盐碱性,而保持系对盐碱较为敏感.     

Carbonylation and Loss-of-Function Analyses of SBPase Reveal Its Metabolic Interface Role in Oxidative Stress, Carbon Assimilation, and Multiple Aspects of Growth and Development in Arabidopsis

Sedoheptulose-1,7-bisphosphatase (SBPase) is a Calvin cycle enzyme and functions in photosynthetic carbon fixation. We found that SBPase was rapidly carbonylated in response to methyl viologen (MV) treatments in detached leaves of Arabidopsis plants. In vitro activity analysis of the purified recombinant SBPase showed that SBPase was carbonylated by hydroxyl radicals, which led to enzyme inactivation in an H(2)O(2) dose-dependent manner. To determine the conformity with carbonylation-caused loss in enzymatic activity in response to stresses, we isolated a loss-of-function mutant sbp, which is deficient in SBPase-dependent carbon assimilation and starch biosynthesis. sbp mutant exhibited a severe growth retardation phenotype, especially for the developmental defects in leaves and flowers where SBPASE is highly expressed. The mutation of SBPASE caused growth retardation mainly through inhibition of cell division and expansion, which can be partially rescued by exogenous application of sucrose. Our findings demonstrate that ROS-induced oxidative damage to SBPase affects growth, development, and chloroplast biogenesis in Arabidopsis through inhibiting carbon assimilation efficiency. The data presented here provide a case study that such inactivation of SBPase caused by carbonyl modification may be a kind of adaptation for plants to restrict the operation of the reductive pentose phosphate pathway under stress conditions.     

Cold-induced activation of brown adipose tissue and adipose angiogenesis in mice

Exposure of humans and rodents to cold activates thermogenic activity in brown adipose tissue (BAT). This protocol describes a mouse model to study the activation of BAT and angiogenesis in adipose tissues by cold acclimation. After a 1-week exposure to 4 °C, adult C57BL/6 mice show an obvious transition from subcutaneous white adipose tissue (WAT) into brown-like adipose tissue (BRITE). The BRITE phenotype persists after continuous cold exposure, and by the end of week 5 BRITE contains a high number of uncoupling protein-1-positive mitochondria, a characteristic feature of BAT. During the transition from WAT into BRITE, the vascular density is markedly increased owing to the activation of angiogenesis. In BAT, cold exposure stimulates thermogenesis by increasing the mitochondrial content and metabolic rate. BAT and the increased metabolic rate result in a lean phenotype. This protocol provides an outstanding opportunity to study the molecular mechanisms that control adipose mass.     

Comparison of Hypoglycemic Activity of Trace Elements Absorbed in Fermented Mushroom of <Emphasis Type="Italic">Coprinus comatus</Emphasis>

The effect of fermented mushroom of Coprinus comatus rich in trace elements, including vanadium, chromium, zinc, magnesium, copper, iron, and nickel, on glycemic metabolism was studied in this paper. Alloxan-induced hyperglycemic mice were used in the study. The blood glucose, glycohemoglobin, and glycogen synthesis of the mice were analyzed, respectively. At the same time, the gluconeogenesis of the normal mice was also determined. After the mice were administered (ig) with C. comatus rich in vanadium (CCRV), the blood glucose and the glycohemoglobin of alloxan-induced hyperglycemic mice decreased (p < 0.05, p < 0.01), glycogen synthesis of alloxan-induced hyperglycemic mice elevated (p < 0.01), the gluconeogenesis of the normal mice was inhibited (p < 0.01), and the sugar tolerance of the normal mice was improved. However, the same result did not occur in other groups. Vanadium at lower doses in combination with C. comatus induced significant effect on glycemic metabolism in mice.