Characteristics of plasmids in multi-drug-resistant Enterobacteriaceae isolated during prospective surveillance of a newly opened hospital in Iraq

Background

Gram-negative multidrug-resistant (MDR) bacteria are major causes of nosocomial infections, and antibiotic resistance in these organisms is often plasmid mediated. Data are scarce pertaining to molecular mechanisms of antibiotic resistance in resource constrained areas such as Iraq.

Methodology/Principal Findings

In this study, all MDR Enterobacteriaceae (n = 38) and randomly selected non-MDR counterparts (n = 41) isolated from patients, healthcare workers and environmental surfaces in a newly opened hospital in Iraq were investigated to characterize plasmids found in these isolates and determine their contribution to antibiotic resistance. Our results demonstrated that MDR E. coli and K. pneumoniae isolates harbored significantly more (≥3) plasmids compared to their non-MDR counterparts, which carried ≤2 plasmids (p<0.01). Various large plasmids (∼52 to 100 kb) from representative isolates were confirmed to contain multiple resistance genes by DNA microarray analysis. Aminoglycoside (acc, aadA, aph, strA/B, and ksgA), β-lactam (bla _TEM1, bla _AMPC, bla _CTX-M-15, bla _OXA-1, bla _VIM-2 and bla _SHV), sulfamethoxazole/trimethoprim (sul/dfr), tetracycline (tet) and chloramphenicol (cat) resistance genes were detected on these plasmids. Additionally, multiple plasmids carrying multiple antibiotic resistance genes were found in the same host strain. Genetic transfer-associated genes were identified on the plasmids from both MDR and non-MDR isolates. Seven plasmid replicon types (FII, FIA, FIB, B/O, K, I1 and N) were detected in the isolates, while globally disseminated IncA/C and IncHI1 plasmids were not detected in these isolates.

Conclusions/Significance

This is the first report of the characteristics of the plasmids found in Enterobacteriaceae isolated following the opening of a new hospital in Iraq. The information provided here furthers our understanding of the mechanisms of drug resistance in this specific region and their evolutionary relationship with other parts of world. The large plasmids, carrying resistance genes and transfer-associated genes, may be potential factors for regional dissemination of antibiotic resistance.     

The membrane proximal disulfides of the EGF receptor extracellular domain are required for high affinity binding and signal transduction but do not play a role in the localization of the receptor to lipid rafts

The EGF receptor is a transmembrane receptor tyrosine kinase that is enriched in lipid rafts. Subdomains I, II and III of the extracellular domain of the EGF receptor participate in ligand binding and dimer formation. However, the function of the cysteine-rich subdomain IV has not been elucidated. In this study, we analyzed the role of the membrane-proximal portion of subdomain IV in EGF binding and signal transduction. A double Cys-->Ala mutation that breaks the most membrane-proximal disulfide bond (Cys600 to Cys612), ablated high affinity ligand binding and substantially reduced signal transduction. A similar mutation that breaks the overlapping Cys596 to Cys604 disulfide had little effect on receptor function. Mutation of residues within the Cys600 to Cys612 disulfide loop did not alter the ligand binding or signal transducing activities of the receptor. Despite the fact that the C600,612A EGF receptor was significantly impaired functionally, this receptor as well as all of the other receptors with mutations in the region of residues 596 to 612 localized normally to lipid rafts. These data suggest that the disulfide-bonded structure of the membrane-proximal portion of the EGF receptor, rather than its primary sequence, is important for EGF binding and signaling but is not involved in localizing the receptor to lipid rafts.     

薇甘菊萎蔫病毒感染对薇甘菊光合特性和4种酶活性的影响

为明确薇甘菊萎蔫病毒(Mikania micrantha wilt virus,MMWV)对薇甘菊(Mikania micrantha)叶片生理生化的影响,探讨了MMWV侵染对薇甘菊叶片超氧化物岐化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)、多酚氧化酶(polyphenol oxidase,PPO)和苯丙氨酸解氨酶(phenylalanine ammonia-lyase,PAL)活性及叶绿素总含量和光合特性的影响.结果表明:MMWV感染薇甘菊16d内叶片的SOD活性均比对照组高,其中第16天达最大值,但接种后第24和32天SOD活性分别比对照低了13.28％和25.37％;POD活性在接种后16-32d均显著高于对照.PPO和PAL变化趋势相似,在接种后第8天这两个酶的活性分别比对照高了77.75％和23.58％,而在第32天分别比对照减少了14.27％和20.53％.随着侵染时间的增加,感病薇甘菊叶片中叶绿素含量减少,叶绿素a/b值逐步降低;同时最大净光合速率(Pmax)和光饱和点(LSP)分别比健康对照减少了32.34％和12.52％,而对暗呼吸速率(Rd),光补偿点(LCP)和表观量子效率(AQY)无显著影响.表明MMWV侵染可减低薇甘菊叶片光合作用的效率.     

The impact of cell-specific absorption properties on the correlation of electron transport rates measured by chlorophyll fluorescence and photosynthetic oxygen production in planktonic algae

Photosynthesis–irradiance (P–E)-curves describe the photosynthetic performance of autotrophic organisms. From these P–E-curves the photosynthetic parameters α-slope, P_max, and E_k can be deduced which are often used to characterize and to compare different organisms or organisms in acclimation to different environmental conditions. Particularly, for in situ-measurements of P–E curves of phytoplankton the analysis of variable chlorophyll fluorescence proved its potential as a sensitive and rapid method. By using Chlorella vulgaris (Trebouxiophyceae), Nannochloropsis salina (Eustigmatophyceae), Skeletonema costatum and Cyclotella meneghiniana (Bacillariophyceae), the present study investigated the influence of cellular bio-optical properties on the correlation of the photosynthetic parameters derived from fluorescence-based P–E-curves with photosynthetic parameters obtained from the measurement of oxygen evolution. It is demonstrated that small planktonic algae show a wide range of cellular absorptivity which was subject to species-specifity, growth stage and environmental conditions, e.g. nutrient limitation. This variability in bio-optical properties resulted in a great deviation of relative electron transport rates (rETRs) from oxygen-based photosynthesis rates. Thus, the photosynthetic parameters α-slope and P_max derived from rETRs strongly depend on the specific cellular absorptivity and cannot be used to compare the photosynthetic performance of cells with different optical properties. However, it was shown that E_k is independent of cellular absorptivity and could be used to compare samples with unknown optical properties.