Nutrition of Myxococcus xanthus FBa and Some of Its Auxotrophic Mutants

A defined medium containing 15 amino acids plus salts was used to study the nutrition of Myxococcus xanthus FBa. The amino acids phenylalanine, leucine, isoleucine, valine, and methionine were essential for growth, whereas glycine, proline, asparagine, alanine, lysine, and threonine stimulated growth. An unusual pattern of requirement was found in the aromatic amino acids. Phenylalanine was essential and served as the precursor of tyrosine. Growth in the absence of tryptophan was adaptive, with cells reaching a growth rate equal to that of controls after a lag of about a week. (14)C-labeled ribose and glucose were not appreciably metabolized. Auxotrophs requiring purines and pyrimidines were isolated and were used to study the fate of externally supplied nucleic acid derivatives. Appropriate mutants could satisfy their requirements with free bases, nucleosides, and nucleotides, and could hydrolyze nucleic acids and use the products. However, studies using (14)C-ribose-labeled uridine (isolated from a Salmonella typhimurium pyrimidine auxotroph) showed that externally supplied nucleic acid derivatives were incorporated almost solely into the nucleic acids of the myxobacters, with little used either for energy-yielding oxidations or other cell anabolism.     

Effects of Low Root Temperature on Ion Uptake and Ion Translocation in Wheat

Roots of wheat seedlings (Triticum aestivum L. cv. Weibulls Starke) were cooled (+1°C) for 24 h while the shoots were kept at 25°C. The treatment induced an increased water deficit in the leaves. Fresh weight, dry weight, and the uptake and distribution of potassium and calcium were measured before and after cooling. Growth, measured both as fresh weight and dry weight increase, was reduced during the cold treatment. Afterwards (at 20°C), growth recovered to nearly pre-stress rates. Analysis of the potassium fluxes in and out of the roots by ⁸⁶Rb techniques showed that influx, and to a lesser extent efflux, were inhibited at low temperature. The result was a net potassium uptake rate of one-third that of unstressed plants. After the cooling period the potassium influx increased to the rate of control plants. The potassium efflux increased to one and one-half times the rate of unstressed wheat so that net uptake was negative. The increase in potassium efflux was explained by a higher permeability of the root cell membranes after cooling. The net uptake of calcium was reduced to one-third by root cooling. Contrary to potassium uptake, calcium uptake increased under post-stress conditions, partly due to a low efflux rate. During root cooling there was a redistribution of dry matter from the leaves down towards the lower part of the shoot. Afterwards the original distribution of dry matter was reestablished. The net flow of potassium and calcium followed a similar pattern as dry matter, suggesting a growth-regulated flow.     

The Deuterator: software for the determination of backbone amide deuterium levels from H/D exchange MS data

Background  

The combination of mass spectrometry and solution phase amide hydrogen/deuterium exchange (H/D exchange) experiments is an effective method for characterizing protein dynamics, and protein-protein or protein-ligand interactions. Despite methodological advancements and improvements in instrumentation and automation, data analysis and display remains a tedious process. The factors that contribute to this bottleneck are the large number of data points produced in a typical experiment, each requiring manual curation and validation, and then calculation of the level of backbone amide exchange. Tools have become available that address some of these issues, but lack sufficient integration, functionality, and accessibility required to address the needs of the H/D exchange community. To date there is no software for the analysis of H/D exchange data that comprehensively addresses these issues.     

Subtle genetic changes enhance virulence of methicillin resistant and sensitive Staphylococcus aureus

Background

Two component lantibiotics, such as the plasmid-encoded lacticin 3147 produced by Lactococcus lactis DPC3147 and staphylococcin C55 produced by Staphylococcus aureus C55, represent an emerging subgroup of bacteriocins. These two bacteriocins are particularly closely related, exhibiting 86% (LtnA1 and C55α) and 55% (LtnA2 and C55β) identity in their component peptides. The aim of this study was to investigate, for the first time for any two component bacteriocins, the significance of the relatedness between these two systems.

Results

So close is this relatedness that the hybrid peptide pairs LtnA1:C55β and C55α:LtnA2 were found to have activities in the single nanomolar range, comparing well with the native pairings. To determine whether this flexibility extended to the associated post-translational modification/processing machinery, the staphylococcin C55 structural genes were directly substituted for their lacticin 3147 counterparts in the ltn operon on the large conjugative lactococcal plasmid pMRC01. It was established that the lacticin LtnA1 post-translational and processing machinery could produce functionally active C55α, but not C55β. In order to investigate in closer detail the significance of the differences between LtnA1 and C55α, three residues in LtnA1 were replaced with the equivalent residues in C55α. Surprisingly, one such mutant LtnA1-Leu21Ala was not produced. This may be significant given the positioning of this residue in a putative lipid II binding loop.

Conclusion

It is apparent, despite sharing striking similarities in terms of structure and activity, that these two complex bacteriocins display some highly dedicated features particular to either system.     

Prevalence of risk factors for cardiovascular disease in Canadians 55 to 74 years of age: results from the Canadian Heart Health Surveys, 1986-1992

BACKGROUND: By 2016, the proportion of Canadians older than 65 years of age will increase to 16%, and there will be an increase in the absolute number of cases of cardiovascular disease in older Canadians. The Canadian Heart Health Surveys database provides information about this population upon which health policy related to cardiovascular disease can be based. This paper presents for the first time population-based data on the risk factors for cardiovascular disease in older Canadians. METHODS: Canadians from all 10 provinces participated in surveys of cardiovascular risk factors; health insurance registries were used as sampling frames. In each province, probability samples of 2200 adults 18 to 74 years old not living in institutions, on reserves or in military camps were asked to participate in interviews and to undergo testing at clinics for major risk factors for cardiovascular disease. RESULTS: A total of 2739 men (response rate 70%) and 2617 women (response rate 66%) aged 55 to 74 years participated in the survey and also provided follow-up clinical measurements at the clinic. Overall, 52% of participants were hypertensive, 26% had isolated systolic hypertension, and 30% had a total blood cholesterol level of 6.2 mmol/L or greater. Rates of current smoking were lower in women than men (17% v. 22%). Overall, 87% of men and 78% of women who were current smokers smoked at least 10 cigarettes per day. Only slightly more than half of participants exercised at least once a week for at least 15 minutes, and almost half had a body mass index of 27 or greater. In only 4% was no major risk factor for cardiovascular disease detected. INTERPRETATION: Significant numbers of older Canadians have one or more major risk factors for cardiovascular disease. Many of these risk factors are amenable to modification.     

Biological affinities and adaptations of Bronze Age Bactrians: IV. A craniometric investigation of Bactrian origins

Discovery of a previously unknown Bronze Age civilization (Oxus civilization) centered on the oases of Central Asia revealed the presence of large, preplanned urban centers immediately above sterile soil. Given the absence of local antecedents, the sudden appearance and proliferation of these Oxus civilization urban centers in the oases of Bactria and Margiana immediately raised the issue of where the inhabitants of these urban centers came from. Three hypotheses have been offered by archaeologists to account for the origins of Oxus civilization populations. These include the early influence model, the late colonization model, and the trichotomy model. Eleven craniometric variables from 12 Aeneolithic and Bronze Age samples, encompassing 657 adults from Central Asia, Iran, and the Indus Valley, are compared to test which if any of these hypotheses are supported by the pattern of phenetic affinities possessed by the Oxus civilization inhabitants of the north Bactrian oasis. Craniometric differences between samples are compared with Mahalanobis generalized distance (d²), and patterns of phenetic affinity are assessed with two types of cluster analysis (WPGMA, neighbor-joining method), multidimensional scaling, and principal coordinates analysis. Results of this analysis provide no support for either the late colonization model or the trichotomy model but do offer some support for the early influence model. Nevertheless, it is clear that the early influence model fails to account for a shift in interregional contacts, perhaps from western China to the north around 2000 bc , that appears to have played a major role in the origins of the Oxus civilization inhabitants of the north Bactrian oasis. Am J Phys Anthropol 108:173–192, 1999. © 1999 Wiley-Liss, Inc.     

大鼠肠内容物细菌基因组DNA提取方法的比较

目的比较两种肠内容物前处理和两种提取方法对清洁级SD大鼠肠内容物细菌基因组DNA提取效率。方法分别选用PBS多次离心漂洗、液氮破细胞两种前处理方法和酚/氯仿抽提、试剂盒过柱法两种提取方法进行组合分析,对4份肠内容物和16份含金黄色葡萄球菌肠内容物进行随机提取。结果大鼠肠内容物细菌基因组DNA含量和纯度测定结果显示,与PBS反复离心相比,液氮研磨前处理能显著提高大鼠肠内容物基因组DNA。荧光定量PCR表明,液氮研磨前处理较PBS反复离心能更好地收集细菌基因组DNA,其Ct值最低。结论研究结果表明,采用液氮研磨试剂盒法在大鼠肠内容物DNA提取中是较为优良的方法,该方法为建立实验动物中微生物的定量PCR检测方法打下了基础。