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961.
Isaac K. O. Cann Kensey R. Amaya Maurice W. Southworth Francine B. Perler 《Applied microbiology》2004,70(5):3158-3162
A genetic selection system that detects splicing and nonsplicing activities of inteins was developed based on the ability to rescue a T4 phage strain with a conditionally inactive DNA polymerase. This phage defect can be complemented by expression of plasmid-encoded phage RB69 DNA polymerase. Insertion of an intein gene into the active site of the RB69 DNA polymerase gene renders polymerase activity and phage viability dependent on protein splicing. The effectiveness of the system was tested by screening for thermosensitive splicing mutants. Development of genetic systems with the potential of identifying protein splicing inhibitors is a first step towards controlling proliferation of pathogenic microbes harboring inteins in essential proteins. 相似文献
962.
Objective: To examine the association between children's overweight status in kindergarten and their academic achievement in kindergarten and first grade. Research Methods and Procedures: The data analyzed consisted of 11, 192 first time kindergartners from the Early Childhood Longitudinal Study, a nationally representative sample of kindergartners in the U.S. in 1998. Multivariate regression techniques were used to estimate the independent association of overweight status with children's math and reading standardized test scores in kindergarten and grade 1. We controlled for socioeconomic status, parent‐child interaction, birth weight, physical activity, and television watching. Results: Overweight children had significantly lower math and reading test scores compared with nonoverweight children in kindergarten. Both groups were gaining similarly on math and reading test scores, resulting in significantly lower test scores among overweight children at the end of grade 1. However, these differences, except for boys’ math scores at baseline (difference = 1.22 points, p = 0.001), became insignificant after including socioeconomic and behavioral variables, indicating that overweight is a marker but not a causal factor. Race/ethnicity and mother's education were stronger predictors of test score gains or levels than overweight status. Discussion: Significant differences in test scores by overweight status at the beginning of kindergarten and the end of grade 1 can be explained by other individual characteristics, including parental education and the home environment. However, overweight is more easily observable by other students compared with socioeconomic characteristics, and its significant (unadjusted) association with worse academic performance can contribute to the stigma of overweight as early as the first years of elementary school. 相似文献
963.
Francine G Smith Alp Sener Raghmeet Basati Isam Abu-Amarah 《Journal of applied physiology》2004,96(1):131-136
Experiments were carried out in conscious, chronically instrumented lambs (n = 8) and young adult sheep (n = 11) to investigate age-dependent renal responses to hemorrhage. Various parameters of renal function were measured for 1 h before and 1 h after either 10% hemorrhage (experiment 1) or 20% hemorrhage (experiment 2). The two experiments were carried out in random order at intervals of 2-5 days. There were no effects of 10-20% hemorrhage on renal plasma flow in either age group. Blood pressure decreased after 20% but not 10% hemorrhage in both age groups. Glomerular filtration rate and filtration fraction decreased after 20% hemorrhage in both age groups, the decrease being greater in lambs than young adult sheep. In response to 20% hemorrhage, urinary flow rate and urinary Na+ excretion rate decreased by 40 min after hemorrhage in young adult sheep but not lambs and remained decreased for 60 min; urinary chloride excretion rate showed a similar response. In lambs but not young adult sheep, free water clearance increased by 20 min after 20% hemorrhage and remained above control at 60 min. Urinary osmolality decreased at 20 min after 20% hemorrhage in young adult sheep but not lambs, returning to control levels by 40 min. These data provide new information that renal responses to hypotensive hemorrhage appear to be developmentally regulated. 相似文献
964.
965.
Marie-Béatrice Bogeat-Triboulot François Bartoli Jean Garbaye Roland Marmeisse Denis Tagu 《Plant and Soil》2004,267(1-2):213-223
Pinus pinaster seedlings were grown in a sandy dune soil either inoculated withHebeloma cylindrosporum or let to natural colonisation. Six months later, half of the seedlings of both treatments were subjected to a 3-week moderate
drought. Root colonisation analysis showed that root tips were colonised to almost 100% independent of the inoculation. DNA
determination of the ectomycorrhizal morphotypes showed that inoculated seedlings were extensively mycorrhized byH. cylindrosporum (more than 75%) whereas non-inoculated seedlings were mycorrhized by the exotic speciesThelephora terrestris (50%) andLaccaria bicolor (30%) and to a lesser extent byH. cylindrosporum (20%). Drought did not affect these frequencies. Total plant biomass was not affected by the mycorrhizal status or by drought
but the root/shoot biomass ratio as well as the root/leaf surface area ratio were much lower in seedlings extensively colonised
byH. cylindrosporum. Root hydraulic conductivity was higher in plants mainly mycorrhized byH. cylindrosporum, showing that this fungus improved the water uptake capacity of the root system as compared toT. terrestris and/orL. bicolor. This positive effect was also found under drought but to a lesser extent.H. cylindrosporum also increased the amount of root-adhering soil as compared to the other fungal symbionts, illustrating the performance of
this association in aggregating sandy soil particles and developing the rhizosheath. The origin of the reduced root hydraulic
resistance byH. cylindrosporum mycorrhization is discussed for the whole path including soil, soil-root interface and root cortex. 相似文献
966.
Roland Ludwig Magdalena Ozga Marcel Z mocky Clemens Peterbauer Klaus D. Kulbe Dietmar Haltrich 《Biocatalysis and Biotransformation》2004,22(2):97-104
An efficient enzymatic bioprocess is described in which lactose, an abundant renewable resource produced by the dairy industry, is completely and efficiently converted with a specific productivity of up to 32 g (kU h)-1 into lactobionic acid, without the formation of any by-products. The key biocatalyst of this new process is the fungal enzyme cellobiose dehydrogenase which oxidizes several β-1,4-linked disaccharides including lactose specifically at position C-1 of the reducing sugar moiety to the corresponding lactones. The electron acceptor employed in this reaction is continuously regenerated with the help of laccase, a H2O-producing, copper-containing oxidase, and therefore has to be added in low, catalytic amounts only. Redox mediators that were successfully employed in this novel process and hence are compatible with the laccase regeneration system include benzoquinone, ABTS, ferricyanide, or ferrocene, amongst others. Factors affecting operational stability of the biocatalysts employed in this process include the redox mediator used, the temperature, and importantly the volumetric gas flow necessary for maintaining the dissolved oxygen tension. Lactobionic acid is a mild and sweet tasting acid with excellent chelating properties. These useful characteristics have lead to a growing number of patents for diverse applications in the food, pharmaceutical and detergent industries. 相似文献
967.
Spangler CM Spangler C Göttle M Shen Y Tang WJ Seifert R Schäferling M 《Analytical biochemistry》2008,381(1):86-93
Adenylyl cyclases catalyze the production of the second messenger cyclic AMP from ATP. Until now, there has been no fluorescent adenylyl cyclase assay known that is applicable to high-throughput screening and kinetic determinations that can directly monitor the turnover of the unmodified substrate ATP. In this study, a fluorescence-based assay is described using the Ca(II)- and calmodulin-dependent adenylyl cyclase edema factor (EF) from Bacillus anthracis and Tb(III)-norfloxacin as probe for the enzyme activity. This assay can be used to study enzyme regulators, allows real-time monitoring of adenylyl cyclase activity, and does not substitute ATP by fluorescent derivatives. These derivatives must be judged critically due to their interference on the activity of enzymes. Furthermore, the new assay makes redundant the application of radioactively labeled substrates such as [α-32P]ATP or fluorescently labeled antibodies such as anti-cyclic AMP. We determined the Michaelis-Menten constant (KM), the v0max value of ATP turnover, and the IC50 values for three inhibitors of EF by this newly developed fluorescent method. 相似文献
968.
Genome-Scale Genotype-Phenotype Matching of Two Lactococcus lactis Isolates from Plants Identifies Mechanisms of Adaptation to the Plant Niche 下载免费PDF全文
Roland J. Siezen Marjo J. C. Starrenburg Jos Boekhorst Bernadet Renckens Douwe Molenaar Johan E. T. van Hylckama Vlieg 《Applied microbiology》2008,74(2):424-436
Lactococcus lactis is a primary constituent of many starter cultures used for the manufacturing of fermented dairy products, but the species also occurs in various nondairy niches such as (fermented) plant material. Three genome sequences of L. lactis dairy strains (IL-1403, SK11, and MG1363) are publicly available. An extensive molecular and phenotypic diversity analysis was now performed on two L. lactis plant isolates. Diagnostic sequencing of their genomes resulted in over 2.5 Mb of sequence for each strain. A high synteny was found with the genome of L. lactis IL-1403, which was used as a template for contig mapping and locating deletions and insertions in the plant L. lactis genomes. Numerous genes were identified that do not have homologs in the published genome sequences of dairy L. lactis strains. Adaptation to growth on substrates derived from plant cell walls is evident from the presence of gene sets for the degradation of complex plant polymers such as xylan, arabinan, glucans, and fructans but also for the uptake and conversion of typical plant cell wall degradation products such as α-galactosides, β-glucosides, arabinose, xylose, galacturonate, glucuronate, and gluconate. Further niche-specific differences are found in genes for defense (nisin biosynthesis), stress response (nonribosomal peptide synthesis and various transporters), and exopolysaccharide biosynthesis, as well as the expected differences in various mobile elements such as prophages, plasmids, restriction-modification systems, and insertion sequence elements. Many of these genes were identified for the first time in Lactococcus lactis. In most cases good correspondence was found with the phenotypic characteristics of these two strains. 相似文献
969.
Type II diabetes mellitus is a disease which is characterized by peripheral insulin resistance coupled with a progressive loss of insulin secretion that is associated with a decrease in pancreatic islet beta-cell mass and the deposition of amyloid in the extracellular matrix of beta-cells, which lead to islet cell death. The principal component of the islet amyloid is a pancreatic hormone called islet amyloid polypeptide (IAPP). High-pressure coupled with FT-IR spectroscopic and AFM studies were carried out to elucidate further information about the aggregation pathway as well as the aggregate structures of IAPP. To this end, a comparative fibrillation study of IAPP fragments was carried out as well. As high hydrostatic pressure (HHP) is acting to weaken or even prevent hydrophobic self-organization and electrostatic interactions, application of HHP has been used as a measure to reveal the importance of these interactions in the fibrillation process of IAPP and its fragments. IAPP preformed fibrils exhibit a strong polymorphism with heterogeneous structures, a large population of which are rather sensitive to high hydrostatic pressure, thus indicating a high percentage of ionic and hydrophobic interactions and loose packing of these species. Conversely, fragments 1-19 and 1-29 are resistant to pressure treatment, suggesting more densely packed aggregate structures with less void volume and strong cooperative hydrogen bonding. Furthermore, the FT-IR data indicate that fragment 1-29 has intermolecular beta-sheet conformational properties different from those of fragment 1-19, the latter exhibiting polymorphic behavior with more disordered structures and less strongly hydrogen bonded fibrillar assemblies. The data also suggest that hydrophobic interactions and/or less efficient packing of amino acids 30-37 region leads to the marked pressure sensitivity observed for full-length IAPP. 相似文献
970.
The actin-ADP-ribosylating Clostridium botulinum C2 toxin consists of the enzymatic component C2I and the binding component C2II. C2II forms heptameric channels involved in translocation of the enzymatic component into the target cell. On the basis of the heptameric toxin channel, we studied functional consequences of mutagenesis of amino acid residues probably lining the lumen of the toxin channel. Substitution of glutamate-399 of C2II with alanine blocked channel formation and cytotoxicity of the holotoxin. Although cytotoxicity and rounding up of cells by C2I were completely blocked by exchange of phenylalanine-428 with alanine, the mutation increased potassium conductance caused by C2II in artificial membranes by about 2-3-fold over that of wild-type toxin. In contrast to its effects on single-channel potassium conductance in artificial membranes, the F428A mutation delayed the kinetics of pore formation in lipid vesicles and inhibited the activity of C2II in promoting (86)Rb (+) release from preloaded intact cells after pH shift of the medium. Moreover, F428A C2II exhibited delayed and diminished formation of C2II aggregates at low pH, indicating major changes of the biophysical properties of the toxin. The data indicate that phenylalanine-428 of C2II plays a major role in conformational changes occurring during pore formation of the binding component of C2II. 相似文献