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171.
In vivo and in vitro effects of thiolactomycin on fatty acid biosynthesis in Streptomyces collinus.
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A stable-isotope assay was used to analyze the effectiveness of various perdeuterated short-chain acyl coenzyme A (acyl-CoA) compounds as starter units for straight- and branched-chain fatty acid biosynthesis in cell extracts of Streptomyces collinus. In these extracts perdeuterated isobutyryl-CoA was converted to isopalmitate (a branched-chain fatty acid), while butyryl-CoA was converted to palmitate (a straight-chain fatty acid). These observations are consistent with previous in vivo analyses of fatty acid biosynthesis in S. collinus, which suggested that butyryl-CoA and isobutyryl-CoA function as starter units for palmitate and isopalmitate biosynthesis, respectively. Additionally, in vitro analysis demonstrated that acetyl-CoA can function as a starter unit for palmitate biosynthesis. Palmitate biosynthesis and isopalmitate biosynthesis in these cell extracts were both effectively inhibited by thiolactomycin, a known type II fatty acid synthase inhibitor. In vivo experiments demonstrated that concentrations of thiolactomycin ranging from 0.1 to 0.2 mg/ml produced both a dramatic decrease in the cellular levels of branched-chain fatty acids and a surprising three- to fivefold increase in the cellular levels of the straight-chain fatty acids palmitate and myristate. Additional in vivo incorporation studies with perdeuterated butyrate suggested that, in accord with the in vitro studies, the biosynthesis of the palmitate from butyryl-CoA decreases in the presence of thiolactomycin. In contrast, in vivo incorporation studies with perdeuterated acetate demonstrated that the biosynthesis of palmitate from acetyl-CoA increases in the presence of thiolactomycin. These observations clearly demonstrate that isobutyryl-CoA is a starter unit for isopalmitate biosynthesis and that either acetyl-CoA or butyryl-CoA can be a starter unit for palmitate biosynthesis in S. collinus. However, the pathway for palmitate biosynthesis from acetyl-CoA is less sensitive to thiolactomycin, and it is suggested that the basis for this difference is in the initiation step. 相似文献
172.
In Upper Three Runs Creek (Aiken, S.C.) and many other environments, less than 1% of bacteria visible microscopically can be cultured. Exploitation of molecular biology techniques has led to development of new methods, such as extraction of nucleic acids from soils or sediments, to study the dominant, nonculturable bacteria. The purpose of this study was to compare three published methods of DNA extraction that fall into two general categories: those in which cells are lysed in sediments (the Ogram and Tsai and methods [A. Ogram, G. S. Sayler, and T. Barkay, J. Microbiol. Methods 7:57-66, 1987; Y. L. Tsai and B. H. Olson, Appl. Environ. Microbiol. 57:1070-1074, 1991]) and those in which cells are removed from sediments prior to lysis (the Jacobsen method [C. S. Jacobsen and O. S. Rasmussen; Appl. Environ. Microbiol. 58:2458-2462, 1992]). DNA yield varied with extraction method; the Ogram method had a significantly higher yield than the other methods. However, DNA extracted via the Ogram method was badly sheared and contained a smaller proportion of eubacterial DNA. The Tsai method was less time consuming than the other methods, but DNA samples were of lower purity. If DNA purity is of paramount concern (as would be the case if PCR was to be performed) and quantity is not important, the Jacobsen method is recommended because of the low concentration of contaminants. If DNA is to be used directly in DNA-DNA hybridizations, the Ogram method is recommended since it gives maximal yields.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
173.
Genetic structure of a lotic population of Burkolderia (Pseudomonas) cepacia. 总被引:3,自引:0,他引:3
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The genetic structure of a population of Burkholderia (Pseudomonas) cepacia isolated from a southeastern blackwater stream was investigated by using multilocus enzyme electrophoresis to examine the allelic variation in eight structural gene loci. Overall, 213 isolates were collected at transect points along the stream continuum, from both the sediments along the bank and the water column. Multilocus enzyme electrophoresis analysis revealed 164 distinct electrophoretic types, and the mean genetic diversity of the entire population was 0.574. Genetic diversity values did not vary spatially along the stream continuum. From a canonical discriminant analysis, Mahalonobis distances (measurements of genetic similarity between populations) revealed significant differences among the subpopulations at the sediment sampling points, suggesting bacterial adaptation to a heterogeneous (or patchy) microgeographical environment. Multilocus linkage disequilibrium analysis of the isolates revealed only limited association between alleles, suggesting frequent recombination, relative to binary fission, in this population. Furthermore, the dendrogram created from the data of this study and the allele mismatch distribution are typical of a population characterized by extensive genetic mixing. We suggest that B. cepacia be added to the growing list of bacteria that are not obligatorily clonal. 相似文献
174.
D. C. Freeman W. A. Turner E. D. McArthur J. H. Graham 《American journal of botany》1991,78(6):805-815
Hybridization between Artemisia tridentata ssp. tridentata and A. t. ssp. vaseyana occurs in a narrow elevational zone along the west face and canyons of the Wasatch Mountains of Utah. Two sites in central Utah (Orem and Salt Creek) were examined. The parental taxa differ in anatomy, morphology, flavonoid and coumarin content, as well as various growth parameters. Hybrids are intermediate for the majority of traits examined, including flavonoid and coumarin composition. Multivariate statistics show that these taxa can be unambiguously identified using simple field measures of morphology. A hybrid index was constructed using principal component analyses on morphological variables. 相似文献
175.
Seed collections from 72 rubber rabbitbrush populations occupying a range of habitats in western North America were incubated at 3 C in the laboratory. Collections from warm desert habitats required less than 2 weeks to achieve 90% relative germination under these conditions, while collections from montane habitats showed delayed germination requiring up to 20 weeks. When 13 representative collections were incubated at constant temperatures from 5 to 30 C, all germinated completely at 30 C within 4 weeks. Collections from warm desert habitats germinated rapidly over the whole range of temperatures. Montane collections sometimes exhibited dormancy at intermediate temperatures (15 and 25 C) even though they were ultimately able to germinate at lower temperatures. Results suggest that dormancy is conditional and temperature-dependent in this species. Chilling the seeds extends the temperature range for germination downward to include the chill temperature itself. Germination response to temperature and its variation as a function of habitat are of apparent adaptive significance, serving to time germination so that the probability of seedling survival is maximized in each habitat. Within populations, response to temperature varied as a function of year of harvest and of within-year harvest date, indicating that germination patterns are probably not under rigid genetic control but represent an integration of genetic and environmental factors. 相似文献
176.
Daniel Q. Li Richard B. Kim Eric McArthur Jamie L. Fleet Robert A. Hegele Baiju R. Shah Matthew A. Weir Amber O. Molnar Stephanie Dixon Jack V. Tu Sonia Anand Amit X. Garg 《PloS one》2016,11(3)
Background
Compared to Caucasians, Chinese achieve a higher blood concentration of statin for a given dose. It remains unknown whether this translates to increased risk of serious statin-associated adverse events amongst Chinese patients.Methods
We conducted a population-based retrospective cohort study of older adults (mean age, 74 years) newly prescribed a statin in Ontario, Canada between 2002 and 2013, where 19,033 Chinese (assessed through a validated surname algorithm) were matched (1:3) by propensity score to 57,099 non-Chinese. This study used linked healthcare databases.Findings
The follow-up observation period (mean 1.1, maximum 10.8 years) was similar between groups, as were the reasons for censoring the observation period (end of follow-up, death, or statin discontinuation). Forty-seven percent (47%) of Chinese were initiated on a higher than recommended statin dose. Compared to non-Chinese, Chinese ethnicity did not associate with any of the four serious statin-associated adverse events assessed in this study [rhabdomyolysis hazard ratio (HR) 0.61 (95% CI 0.28 to 1.34), incident diabetes HR 1.02 (95% CI 0.80 to 1.30), acute kidney injury HR 0.90 (95% CI 0.72 to 1.13), or all-cause mortality HR 0.88 (95% CI 0.74 to 1.05)]. Similar results were observed in subgroups defined by statin type and dose.Conclusions
We observed no higher risk of serious statin toxicity in Chinese than matched non-Chinese older adults with similar indicators of baseline health. Regulatory agencies should review available data, including findings from our study, to decide if a change in their statin dosing recommendations for people of Chinese ethnicity is warranted. 相似文献177.
Anusha Ande Carole McArthur Leo Ayuk Charles Awasom Paul Ngang Achu Annette Njinda Namita Sinha P. S. S. Rao Marisela Agudelo Anantha Ram Nookala Stephen Simon Anil Kumar Santosh Kumar 《PloS one》2015,10(4)
Mild-to-moderate tobacco smoking is highly prevalent in HIV-infected individuals, and is known to exacerbate HIV pathogenesis. The objective of this study was to determine the specific effects of mild-to-moderate smoking on viral load, cytokine production, and oxidative stress and cytochrome P450 (CYP) pathways in HIV-infected individuals who have not yet received antiretroviral therapy (ART). Thirty-two human subjects were recruited and assigned to four different cohorts as follows: a) HIV negative non-smokers, b) HIV positive non-smokers, c) HIV negative mild-to-moderate smokers, and d) HIV positive mild-to-moderate smokers. Patients were recruited in Cameroon, Africa using strict selection criteria to exclude patients not yet eligible for ART and not receiving conventional or traditional medications. Those with active tuberculosis, hepatitis B or with a history of substance abuse were also excluded. Our results showed an increase in the viral load in the plasma of HIV positive patients who were mild-to-moderate smokers compared to individuals who did not smoke. Furthermore, although we did not observe significant changes in the levels of most pro-inflammatory cytokines, the cytokine IL-8 and MCP-1 showed a significant decrease in the plasma of HIV-infected patients and smokers compared with HIV negative non-smokers. Importantly, HIV-infected individuals and smokers showed a significant increase in oxidative stress compared with HIV negative non-smoker subjects in both plasma and monocytes. To examine the possible pathways involved in increased oxidative stress and viral load, we determined the mRNA levels of several antioxidant and cytochrome P450 enzymes in monocytes. The results showed that the levels of most antioxidants are unaltered, suggesting their inability to counter oxidative stress. While CYP2A6 was induced in smokers, CYP3A4 was induced in HIV and HIV positive smokers compared with HIV negative non-smokers. Overall, the findings suggest a possible association of oxidative stress and perhaps CYP pathway with smoking-mediated increased viral load in HIV positive individuals. 相似文献
178.
179.
The effect of the iron-binding glycoprotein lactoferrin (LF) on the in vitro primary antibody response of mouse splenic cells to a T-lymphocyte-dependent antigen, sheep erythrocytes (SE), and a T-lymphocyte-independent antigen, trinitrophenolated Brucella abortus, was examined. Both iron-saturated and native LF (8% saturated) at 10?10 to 10?6M concentrations but not transferrin (an iron-binding glycoprotein similar to LF) significantly (P < 0.05) decreased the number of direct (IgM) plaque-forming cells (PFC) to SE and trinitrophenol (TNP) as determined by the hemolytic plaque assay. LF was equally effective in decreasing the PFC response to TNP in T-lymphocyte-depleted splenic cell cultures. Concentrations of LF which decreased the PFC response were noncytotoxic. A 1-hr exposure of splenic cells to LF at the beginning of the 5-day culture period but not 1 hr prior to assaying for PFC, or exposure of isolated macrophage-rich but not lymphocyte-rich populations to LF prior to reconstitution resulted in a significant decrease in the anti-SE response. These data suggest that LF which is synthesized and released by polymorphonuclear leukocytes and present in inflammatory lesions may play a role in modulating the antibody response through an effect on the macrophage. 相似文献
180.
N K Clapp A H McArthur R L Carson M A Henke O C Peck J D Wood 《Laboratory animal science》1987,37(2):217-219
Endoscopic visualization and biopsy have been performed under anesthesia in more than 65 tamarins and marmosets to study the pathogenesis of colitis and cancer of the colon. This procedure allows examination of the large bowel from the anus to the cecum and has been repeated at 2-6 month intervals with few complications. However, care must be exercised not to perforate the colon. Successful use of this technique will permit study of the pathogenesis of colonic diseases throughout the life of the animal and should provide cause-effect information about colitis and colon cancer in tamarins that may apply to the human diseases. 相似文献