Novel Biotransformations of 7-Ethoxycoumarin by Streptomyces griseus

Biotransformation of 7-ethoxycoumarin by Streptomyces griseus resulted in the accumulation of two metabolites which were isolated and identified as 7-hydroxycoumarin and 7-hydroxy-6-methoxycoumarin. A novel series of biotransformation reactions is implicated in the conversion of the ethoxycoumarin substrate to these products, including O-deethylation, 6-hydroxylation to form a 6,7-dihydroxycoumarin catechol, and subsequent O-methylation. Either 7-hydroxycoumarin or 6,7-dihydroxycoumarin was biotransformed to 7-hydroxy-6-methoxycoumarin by S. griseus. Trace amounts of the isomeric 6-hydroxy-7-methoxycoumarin were detected when 6,7-dihydroxycoumarin was used as the substrate. Efforts to obtain a cell-free catechol-O-methyltransferase enzyme system from S. griseus were unsuccessful. However, [methyl-¹⁴C]methionine was used with cultures of S. griseus to form 7-hydroxy-6-[¹⁴C]methoxycoumarin.     

Involvement of an Inducible Cytochrome P-450 in Precocene II Oxidation by Streptomyces Griseus

Streptomyces griseus oxidizes the insecticide precocene II to its cis- and trans-dihydrodiols and 3-chromenol after growth on an enriched medium containing soybean flour. Oxidation of precocene II is dependent on the level of cytochrome P-450 in this organism. Extracts of cells grown on media lacking soybean flour were devoid of cytochrome P-450 and could not oxidize precocene II. In an in vitro reconstituted system containing NADPH, spinach ferredoxin reductase, spinach ferredoxin and ammonium sulfate fractions enriched in cytochrome P-450, precocene II was oxidized to its dihydrodiols. An aerial mycelium-negative variant of S. griseus (AMY mutant), that was unable to elicit cytochrome P-450 when grown on soybean flour-enriched medium, failed to oxidize precocene II.     

A unique genomic sequence in the Wolf-Hirschhorn syndrome [WHS] region of humans is conserved in the great apes

The Wolf-Hirschhorn syndrome (WHS) is caused by a partial deletion in the short arm of chromosome 4 band 16.3 (4p16.3). A unique-sequence human DNA probe (39 kb) localized within this region has been used to search for sequence homology in the apes' equivalent chromosome 3 by FISH-technique. The WHS loci are conserved in higher primates at the expected position. Nevertheless, a control probe, which detects alphoid sequences of the pericentromeric region of humans, is diverged in chimpanzee, gorilla, and orangutan. The conservation of WHS loci and divergence of DNA alphoid sequences have further added to the controversy concerning human descent.     

BRCA1 maps proximal to D17S579 on chromosome 17q21 by genetic analysis

Previous studies have demonstrated linkage between early-onset breast cancer and ovarian cancer and genetic markers on chromosome 17q21. These markers define the location of a gene (BRCA1) which appears to be inherited as an autosomal dominant susceptibility allele. We analyzed five families with multiple affected individuals for evidence of linkage to the BRCA1 region. Two of the five families appear to be linked to BRCA1. One apparently linked family contains critical recombinants, suggesting that the gene is proximal to the marker D17S579 (Mfd188). These findings are consistent with the maximum-likelihood position estimated by the Breast Cancer Linkage Consortium and with recombination events detected in other linked families. Linkage analysis was greatly aided by PCR-based analysis of paraffin-embedded normal breast tissue from deceased family members, demonstrating the feasibility and importance of this approach. One of the two families with evidence of linkage between breast cancer and genetic markers flanking BRCA1 represents the first such family of African-American descent to be reported in detail.     

Phytopathogenic bacteria utilize host glucose as a signal to stimulate virulence through LuxR homologues

Chemical signal-mediated biological communication is common within bacteria and between bacteria and their hosts. Many plant-associated bacteria respond to unknown plant compounds to regulate bacterial gene expression. However, the nature of the plant compounds that mediate such interkingdom communication and the underlying mechanisms remain poorly characterized. Xanthomonas campestris pv. campestris (Xcc) causes black rot disease on brassica vegetables. Xcc contains an orphan LuxR regulator (XccR) which senses a plant signal that was validated to be glucose by HPLC-MS. The glucose concentration increases in apoplast fluid after Xcc infection, which is caused by the enhanced activity of plant sugar transporters translocating sugar and cell-wall invertases releasing glucose from sucrose. XccR recruits glucose, but not fructose, sucrose, glucose 6-phosphate, and UDP-glucose, to activate pip expression. Deletion of the bacterial glucose transporter gene sglT impaired pathogen virulence and pip expression. Structural prediction showed that the N-terminal domain of XccR forms an alternative pocket neighbouring the AHL-binding pocket for glucose docking. Substitution of three residues affecting structural stability abolished the ability of XccR to bind to the luxXc box in the pip promoter. Several other XccR homologues from plant-associated bacteria can also form stable complexes with glucose, indicating that glucose may function as a common signal molecule for pathogen–plant interactions. The conservation of a glucose/XccR/pip-like system in plant-associated bacteria suggests that some phytopathogens have evolved the ability to utilize host compounds as virulence signals, indicating that LuxRs mediate an interkingdom signalling circuit.     

Sub-Cellular Distribution of Zinc in Different Vegetative Organs and Their Contribution to Grains Zinc Accumulation in Rice Under Different Nitrogen and Zinc Supply

Journal of Plant Growth Regulation - Zinc is an important micronutrient for the growth and development of human body and plants. Proper use of nitrogen fertilizer and foliar application of Zn have...     

β-桧木醇联合替加环素对tet(X4)阳性大肠杆菌的体外协同抗菌作用

质粒介导tet(X4)基因的出现和流行,严重影响替加环素(tigecycline)对临床多重耐药细菌感染的治疗效果,急需寻找有效的佐剂遏制替加环素耐药性。本研究采用微量棋盘稀释法、时间-杀菌曲线测定β-桧木醇(β-thujaplicin)和替加环素的体外联合抗菌表征,通过测定细菌细胞膜通透性、细菌胞内活性氧(reactive oxygen species,ROS)含量、铁含量以及替加环素含量等指标,来探究β-桧木醇和替加环素联合使用对tet(X4)基因阳性大肠杆菌(Escherichia coli)的抗菌作用机制。结果表明,β-桧木醇联合替加环素对tet(X4)基因阳性大肠杆菌具有体外协同抗菌效果,且β-桧木醇在抗菌作用浓度范围内无显著溶血性和细胞毒性。机制研究发现,β-桧木醇能显著增大细菌细胞膜的通透性,降低细菌胞内铁含量,干扰铁稳态,诱导细胞内ROS显著增加,从而发挥抗菌效果。进一步研究发现,β-桧木醇可通过干扰细菌铁代谢和增大细菌细胞膜通透性,协同增强替加环素的抗菌效果。本研究结果为β-桧木醇联合替加环素治疗tet(X4)基因阳性大肠杆菌感染提供了理论和实践基础。     

Species composition,similarity, and structure of mayan homegardens in tixpeual and tixcacaltuyub,yucatan, Mexico

We studied species composition, similarity, and structure of homegardens in two Yucatecan Maya communities, Tixpeual and Tixcacaltuyub, Yucatan, Mexico. The number of gardens sampled per village was 20 and 22; total area sampled was very similar, 45,265 m² and 40,150 m²; the number of trees and shrubs present was 5651 and 5603; and number of species was 135 and 133, respectively. Diversity was low for both sites (H′= 1.6), as were the correlation coefficients (r) for the species-area and individuals-area correlations. The relatively low values obtained for the structural parameters reflect the random pattern of plant incorporation to the gardens, the variability in the proportion of constantly used and not constantly used garden area, and a certain uniformity in the number of species used and number of individuals present, and the relationship between these parameters and garden size. All these reflect the uniqueness of each homegarden, which depends upon the cultural background of the owner. We noticed a trend towards a change in homegarden structure and function in response to the modernization process. Homegardens in villages in the outskirts of cities tend to have more ornamental species and commercial fruit plants than homegardens in isolated villages.