Simple estimate of ureides in soybean tissue

Extracts of soybean tissue are treated with an acidic cation-exchange resin to remove amino acids. The sample is treated with hypochlorite, pH 4.0, to convert the amide from allantoic acid or allantoin to products that react with alkaline phenol to form indophenol. The procedure is inexpensive, sensitive, and rapid. The method may be adapted for automated analysis with the Technicon autoanalyzer.     

二叠纪菊石属Shengoceras的特大型标本在广西的发现

当前研究标本壳体之大,在此类菊石中是罕见的,而且它的缝合线保存亦十分完美.标本体室虽已剥落,仅存留气室,但其壳径仍达101mm,推测完整壳体的壳径至少在150mm以上.标本系由我所张治平一年前得自广西柳州市.根据他所提供的地点及标本本身的特征,推断它可能来自广西柳江、来宾和武宣县一带二叠系的孤峰组.壳形及缝合线表明它即赵金科(1955)所记录、嗣后Ruzhencev(1956)更名为"孤峰菊石( Ku.fengoceras )”的一类化石,只是赵氏记述的标本壳径最大者仅为当前标本的一半左右.     

A gene-driven ENU-based approach to generating an allelic series in any gene

N-ethyl-N-nitrosourea (ENU) introduces mutations throughout the mouse genome at relatively high efficiency. Successful high-throughput phenotype screens have been reported and alternative screens using sequence-based approaches have been proposed. For the purpose of generating an allelic series in selected genes by a sequence-based approach, we have constructed an archive of over 4000 DNA samples from individual F₁ ENU-mutagenized mice paralleled by frozen sperm samples. Together with our previously reported archive, the total size now exceeds 6000 individuals. A gene-based screen of 27.4 Mbp of DNA, carried out using denaturing high-performance liquid chromatography (DHPLC), found a mutation rate of 1 in 1.01 Mbp of which 1 in 1.82 Mbp were potentially functional. Screening of whole or selected regions of genes on subsets of the archive has allowed us to identify 15 new alleles from 9 genes out of 15 tested. This is a powerful adjunct to conventional mutagenesis strategies and has the advantage of generating a variety of alleles with potentially different phenotypic outcomes that facilitate the investigation of gene function. It is now available to academic collaborators as a community resource.     

Novel phenotypes identified by plasma biochemical screening in the mouse

We used ENU mutagenesis in the mouse for the rapid generation of novel mutant phenotypes for both gene function studies and use as new animal models of human disease (Nolan et al. 2000b). One focus of the program was the development of a blood biochemistry screen. At 8-12 weeks of age, approximately 300 ml of blood was collected from F1 offspring of ENU mutagenized male mice. This yielded approximately 125 ml of plasma, used to perform a profile of 17 standard biochemical tests on an Olympus analyzer. Cohorts of F1 mice were also aged and then retested to detect late onset phenotypes. In total, 1,961 F1s were screened. Outliers were identified by running means and standard deviations. Of 70 mice showing consistent abnormalities in plasma biochemistry, 29 were entered into inheritance testing. Of these, 9 phenotypes were confirmed as inherited, 10 found not to be inherited, and 10 are still being tested. Inherited mutant phenotypes include abnormal lipid profiles (low total and HDL cholesterol, high triglycerides); abnormalities in bone and liver metabolism (low ALP, high ALP, high ALT, and AST); abnormal plasma electrolyte levels (high sodium and chloride); as well as phenotypes of interest for the study of diabetes (high glucose). The gene loci bearing the mutations are currently being mapped and further characterized. Our results have validated our biochemical screen, which is applicable to other mutagenesis projects, and we have produced a new set of mutants with defined metabolic phenotypes.     

ReleasingAmblyseius spp. [Acarina: Phytoseiidae] to controlThrips tabaci [Thysanoptera: Thripidae] on cabbage

Two species of predaceous mites,Amblyseius barkeri (Hughes) [=A. mckenziei Schuster & Pritchard] andA. cucumeris (Oudemans) [Acarina: Phytoseiidae] were evaluated as potential biological control agents forThrips tabaci Lindeman [Thysanoptera: Thripidae] on cabbage.A. barkeri colonized cabbage heads in preliminary trials. Field releases of different numbers of mites per plant during 1987 showed thatA. barkeri reduced the number of thrips in cabbage heads at harvest, and the reduction was proportional to the number of mites released, but thrips damage was unaffected. ReleasingA. cucumeris at different times during 1988, but releasing the same number of mites each time, we found that earlier releases resulted in fewer thrips and less damage at harvest; these relationships were not present, however, in plots treated with pyrethroid insecticides. We conclude that inoculative release ofAmblyseius spp. is a potentially useful thrips management strategy, but improvements in release timing and strategy will be required to provide commercially acceptable control.      

Isoform-specific O-glycosylation by murine UDP-GalNAc:polypeptide N- acetylgalactosaminyltransferase-T3, in vivo

Multiple isoforms of UDP-GalNAc:polypeptide N-acetylgalactosaminyl- transferase (ppGaNTase) have been cloned and expressed from a variety of organisms. In general, these isoforms display different patterns of tissue-specific expression, but exhibit overlapping substrate specificities, in vitro . A peptide substrate, derived from the sequence of the V3 loop of the HIV gp120 protein (HIV peptide), has previously been shown to be glycosylated in vitro exclusively by the ppGaNTase-T3 (Bennett et al. , 1996). To determine if this isoform- specificity is maintained in vivo , we have examined the glycosylation of this substrate when it is expressed as a reporter peptide (rHIV) in a cell background (COS7 cells) which lacks detectable levels of the ppGaNTase-T3. Glycosylation of rHIV was greatly increased by coexpression of a recombinant ppGaNTase-T3. Overexpression of ppGaNTase- T1 yielded only partial glycosylation of the reporter. We have also determined that the introduction of a proline residue at the +3 position flanking the potential glycosylation site eliminated ppGaNTase- T3 selectivity toward rHIV observed both in vivo and in vitro .      

Relationship between water content and afterripening in red rice

Reactions regulating seed dormancy can proceed at water contents which are probably too low to permit metabolic activity. The loss of dormancy via afterripening of red rice. ( Oryza sativa L.) seeds was examined as a representative case. Equilibration of seeds to various moisture contents showed that afterripening was most rapid at 6–14% moisture content (dry weight basis). Afterripening did not occur at > 18% moisture content and was severely inhibited at < 5% moisture content. Seed viability was greater than 95% for all treatments. Utilization of moisture isotherms to calculate water-binding enthalpy values identified the optimal afterripening range as approximately the boundary between water-binding region 1 and region 2. From these findings, it is suggested that afterripening may involve some oxidative reactions which are inhibited at lower water contents by the rising free-energy and at the higher side by metabolic reactions.     

A 2x folic acid treatment affects epigenetics and dendritic spine densities in SHSY5Y cells

Many diseases are now associated with aberrant epigenetics and gene expression changes. Epigenetics can be modified by factors like diet. One dietary factor, folic acid, is consumed in various forms including supplements, energy drinks, and fortified grains. It was hypothesized high levels of folic acid would affect gene expression and enzyme activity of chromatin modifying enzymes as well as dendritic spine densities in a commonly utilized neuron model, the SHSY5Y cell. Decreased MBD2 and MECP2 were discovered upon treatment of SHSY5Y cells with a 2x folic acid dose. Corresponding decreases in dendritic spines were apparent in the 2x folic acid treated cells as well. Activity of DNMTs and H3K4 HMTs was altered. Further, H3K4me1, H3K4me3, H3K9Ac, and global DNA methylation were decreased in the 2x folic acid treated cells. Further studies are warranted to determine if the effects of excess folic acid are detrimental to organismal physiology.