Plasmid-mediated resistance to fosfomycin in Staphylococcus epidermidis

Staphylococcus epidermidis strain BM2641, isolated from a patient, was resistant to penicillin G, methicillin, aminoglycosides, chloramphenicol, macrolide, lincosamide and streptogramin B-type (MLS) antibiotics, and to high levels of fosmycin. Resistance to forsfomycin and/or to MLS was lost at low frequencies either spontaneously or after curing with novobiocin. The plasmid DNA from BM2641 and its cured derivatives was purified, analyzed by agarose gel electrophoresis and transferred to a nitrocellulose sheet. Comparative analysis of the resistance phenotypes with the plasmid content of the strains indicated that fosfomycin and MLS resistance were encoded by plasmids pIP1842 (2.5 kb) and pIP1843 (2.6 kb), respectively. Southern hybridization with a probe specific for gene fosA of Serratia marcescens showed that the fosfomycin resistance determinant in Staphylococcus is not homologous to that of Gram-negative bacteria.     

Replication of pSC101: effects of mutations in the E. coli DNA binding protein IHF

Summary We have shown that the plasmid pSC101 is unable to be maintained in strains of E. coli carrying deletions in the genes himA and hip which specify the pleitropic heterodimeric DNA binding protein, IHF. We show that this effect is not due to a modulation of the expression of the pSC101 RepA protein, required for replication of the plasmid. Inspection of the DNA sequence of the essential replication region of pSC101 reveals the presence of a site, located between the DnaA binding-site and that of RepA, which shows extensive homology with the consensus IHF binding site. The proximity of the sites suggests that these three proteins, IHF, DnaA, and RepA may interact in generating a specific DNA structure required for initiation of pSC101 replication.     

Static recipient cells as reservoirs of antibiotic resistance during antibiotic therapy

How does taking the full course of antibiotics prevent antibiotic resistant bacteria establishing in patients? We address this question by testing the possibility that horizontal/lateral gene transfer (HGT) is critical for the accumulation of the antibiotic-resistance phenotype while bacteria are under antibiotic stress. Most antibiotics prevent bacterial reproduction, some by preventing de novo gene expression. Nevertheless, in some cases and at some concentrations, the effects of most antibiotics on gene expression may not be irreversible. If the stress is removed before the bacteria are cleared from the patients by normal turnover, gene expression restarts, converting the residual population to phenotypic resistance. Using mathematical models we investigate how static recipients of resistance genes carried by plasmids accumulate resistance genes, and how specifically an environment cycling between presence and absence of the antibiotic uniquely favors the evolution of horizontally mobile resistance genes. We found that the presence of static recipients can substantially increase the persistence of the plasmid and that this effect is most pronounced when the cost of carriage of the plasmid decreases the cell's growth rate by as much as a half or more. In addition, plasmid persistence can be enhanced even when conjugation rates are as low as half the rate required for the plasmid to persist as a parasite on its own.     

CO2倍增条件下长白赤松幼苗土壤CO2廓线的动态

采用固定在土壤中的气井系统 ,监测土壤剖面的CO2 动态及其与长白赤松 (Pinussylvestrisvar.sylvestriformis(Takenouchi)ChengetC .D .Chu)幼苗根系发展之间的关系。实验研究共设 4种CO2 处理 ,分别是环境CO2 浓度 ,无苗 ;CO2 为 70 0 μmol/mol,无苗 ;环境CO2 浓度 ,有苗 ;CO2 为 70 0 μmol/mol,有苗。通过对土壤剖面CO2 气体的同步采集与分析表明 :土壤CO2 廓线与幼苗根系的生物活性密切相关。在土壤表面及壤土和沙土的边界层中 ,根系分布密集 ,根系的呼吸作用对那两个土层CO2 贡献大 ;随着幼苗的季节生长 ,与环境CO2 浓度比较 ,CO2 倍增将导致土壤剖面上CO2 浓度最大区域由表面向壤土和沙土边界层的转移。本文采用的气井系统提供了一种对土壤无破坏、经济、简单并且能够用于监测幼苗地下过程的廓线研究方法。     

Site-specific integration and excision of pMEA100 in Nocardia mediterranei

Summary Nocardia mediterranei strain LBG A3136 contains the 23.7 kb element pMEA100 in a chromosomally integrated form as well as in the free state (Moretti et al. 1985). The integrated form of this element can be excised precisely from the Nocardia chromosome without any accompanying rearrangements in flanking chromosomal DNA. After transfer into plasmid-free mutant strains, pMEA100 reintegrates site specifically into its original chromosomal locus. The exact mapping of the pMEA100 integration site was accomplished by restriction analysis and DNA sequencing. The attachment site of pMEA100, the junctions of its integrated form and plasmid-free chromosomal DNA of N. mediterranei contain an identical 47 bp long sequence which is probably required for site-specific recombination connected with integration and excision of pMEA100. Only one such sequence was found in the chromosome of pMEA100-free N. mediterranei derivatives as suggested by the single integration locus.     

Studying multiple protein profiles over time to assess biomarker validity

Protein profile analysis is increasingly used for identification of disease biomarkers. The approaches vary from surface-enhanced laser desorption/ionization to protein arrays. Newer platforms are constantly being developed. Almost all are based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and are often coupled with sophisticated software tools. Protein profiling has been applied to a variety of samples including plasma, urine, cerebrospinal fluid, saliva and solid tissue. This article focuses on those instances where it is possible to obtain sequential samples from the same individual. In the authors use of a profile method, many protein changes with highly significant correlations to disease have been found. The main challenge lies in the validation of the marker to demonstrate its adequacy for use in the clinical setting. The latter requires a methodology that is robust and amenable to high-throughput. One problem is that interindividual variability among the healthy population can mask major changes that occur on an intraindividual basis. Often, a large change for an individual may remain within the range of healthy individuals. Thus, one strategy to optimize biomarker discovery is to examine serial samples from a given individual, where a disease biomarker is established by comparison with the individual’s own baseline sample. The focus of this review is to illustrate the principle and value of serial protein profiling using a rapid protein extraction method.     

The bacterial replication initiator DnaA. DnaA and oriC,the bacterial mode to initiate DNA replication

The initiation of replication is the central event in the bacterial cell cycle. Cells control the rate of DNA synthesis by modulating the frequency with which new chains are initiated, like all macromolecular synthesis. The end of the replication cycle provides a checkpoint that must be executed for cell division to occur. This review summarizes recent insight into the biochemistry, genetics and control of the initiation of replication in bacteria, and the central role of the initiator protein DnaA.     

Physiological consequences of an altered flow regime on Alabama bass (Micropterus henshalli)

There are numerous studies on the effects of dams on aquatic biota, yet relatively little is known about whether hydropeaking activities cause physiological change in fish. Using Alabama bass (Micropterus henshalli) as a model, we evaluated whether hydropeaking in a regulated river altered glucocorticoid stress responsiveness relative to fish from an unregulated tributary. Blood samples were collected at the time of capture (baseline) and then collected again after a 1‐hr period of confinement (response). Leukocyte profiles (blood smears) were created and plasma was extracted to assess plasma cortisol levels and neutrophils and lymphocyte (N:L) ratios, between sites and times to evaluate differences between sites and the two sampling periods. Baseline cortisol levels were higher in fish collected from the regulated river compared to those from unregulated site, but response levels of cortisol were similar between sites. Baseline and response level N:L ratios did not differ between sites. High baseline levels of cortisol suggested that fish exposed to regulated flows expressed an altered stress response and were likely in an allostatic state, i.e., attempting to acclimate. Further research is needed to understand how altered stress responses due to hydropeaking flows may be affecting fish.     

Plasmid rolling circle replication and its control

Abstract This review summarises current information on rolling circle replicating plasmids originally isolated from Gram-positive bacteria with a low guanine and cytosine content in their DNA. It focuses on the peculiar biological features of these small, high copy number plasmids that replicate via an asymmetric RC mechanism. The regulation of plasmid copy number is also discussed.