Evaluation of the Frequency of Methicillin-Resistant Staphylococs Isolated from Nose of Nursing Personnel of Hajar Hospital of Shahrekord

Background and Objectives: Methicillin-resistant Staphylococcus strains, as one of the most important etiologic agents of nosocomial infections are of particular importance due to their ability to create potential cross-resistance to all beta-lactam agents. Nasal carriers in hospital staff are supposed to be the main sources of nosocomial infection. This study was conducted aiming at determining the frequency of methicillin-resistant Staphylococcus strains isolated from nose of the personnel of Hadjar Hospital of Shahrekord. Methods: In this descriptive study, nose swabs were collected from 204 personnel of the Hajar hospital of Shahrekord. At first, the nasal swab specimens were cultured on mannitol salt agar (MSA) and TSB. Then, the Staphylococcus strains were isolated and identified using standard microbiological methods, including catalase, coagulase, DNase, and mannitol fermentation tests. In continue, agar screen method was used to determine the susceptibility of the isolated methicillin-resistant Staphylococcus strains. The results were statistically analyzed using chi-square and Fisher’s exact tests. Results: According to the results obtained, 23 of 52 (44%) Staphylococcus aureus strains and 70 strains of 152 (46%) coagulase-negative staphylococcus strains were resistant to methicillin, using agar screen method. The highest percentage of Staphylococcus aureus carriers was isolated from the staff of infectious ward and from the experienced staff (6-10 years) of a special ward. Also, there was no significant relationship between personnel's work experience in the special ward or their workplace and being a carrier. Conclusion: The results of this study demonstrated that the frequency of methicillin-resistant Staphylococcus strains is considerable in the personnel of Hajar Hospital. Therefore, considering the risk of its resulting epidemics in nosocomial infections among hospital's personnel, it seems necessary to detect carriers to control and prevent nosocomial infections

Comparison of Agar screen and duplex-PCR methods in determination of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from nasal carriage

Methicillin-resistant Staphylococcus aureus strains (MRSA) have become a serious health issue in engendering nosocomial infections. Due to the heterogeneity of this type of resistance, the conventional antibiotic susceptibility tests may fail to detect MRSA strains. The purpose of this research was to compare the phenotypic agar screen method with polymerase chain reaction (PCR) for detection of MRSA strains isolated from the nasal samples of hospital personnel. Totally, 52 coagulase positive S. aureus strains were isolated from nasal samples of 204 hospital personnel of Hajar Hospital affiliated to Shahrekord University of Medical Sciences. Susceptibility to oxacillin in the strains was evaluated by the phenotypic agar screen method. The presence of the methicillin resistance gene, mec A, was studied through duplex PCR method. The results of both methods were compared and the sensitivity and specificity of the methods were determined. Totally, 23 out of the 52 isolated S. aureus (44%) were phenotypically resistant to oxacillin, but 27 (52%) carried mecA gene. The sensitivity and specificity of the phenotypic agar screen method for determination of MRSA strains were found to be 81.5 and 96%, respectively. As compared to duplex PCR, oxacillin agar screen method is a simple, inexpensive, and practical phenotypic method with relatively low false positive results and thus may be suitable for verification of suspicious MRSA strains. However, for the relatively high false negative results, it may not be recommended for the primary screening of MRSA strains from the nasal samples of healthy carriers working at hospitals

Comparison of agar screen and duplex-PCR in determination of methicillin resistant Staphylococcus aureus (MRSA) strains isolated from nose of personnel in Hajar hospital of Shahre-kord, 2007

Introduction: Methicillin resistant staphylococcus aureus strains are the most important agents of nosocomial infections. The conventional antibiotic susceptibility methods such as disk diffusion are not suitable for detection of these strains due to their heteroresistancy. Therefore, in this study, agar screen and duplex-PCR were compared in determination of methicillin resistant Staphylococcus aureus (MRSA) strains isolated from nose of personnel in Hajar hospital of Shahre-kord, 2007. Materials and Methods: In this experimental study a total of 204 nasal swabs from personnel of Hajar hospital over a period of 6 months were collected. The specimens were cultured on mannitol salt agar for primary isolation and identification of Staphylococcus aureus strains and their susceptibility pattern to oxacillin was assessed using agar screen method. Finally, using duplex PCR, the isolates were tested for the presence of mecA gene. Results were compared and sensitivity and specificity of the method was determined. Results: In this study, 23 of the 52 (44%) Staphylococcus aureus isolates were resistant to oxacillin using agar screen method. However, mecA gene was detected in 27 of the 52 strains (52%). Our results showed that the sensitivity and specificity of agar screen method in determination of MRSA strains were 81.5% and 96%, respectively comparing with PCR. Conclusion: Oxacillin agar screen, comparing PCR, is an inexpensive, applied and phenotypical method with low false positive and suitable for screening of MRSA. However, due to its relatively high false negative results is not appropriate for screening of MRSA strains isolated from hospital-employed nasal carriers

Recording temporal data with minutes resolution into DNA

Recording complex biological signals is a crucial application of synthetic biology and essential for a deeper understanding of biological processes. An ideal “biorecorder” would have the ability to record biological signals over a wide spatial distribution of cells with high temporal resolution. However, the genetically encoded biorecording tools available have very good spatial resolution (cellular level), but currently rely on turning on and off transcription and translation of a protein (e.g., Cas9 or a recombinase) to record the biological signal, making their temporal resolution on the order of hours. Here we introduce a DNA polymerase based biorecorder that can record cationic concentration fluctuations into DNA sequence with a resolution of ~1 minute. We use a template independent DNA polymerase; terminal deoxynucleotidyl transferase (TdT) that randomly incorporates bases onto a single strand of DNA. The preference of base incorporated by TdT changes with the concentration of cations in TdT’s environment. Therefore, by analyzing a strand of DNA that was extended in fluctuating cation concentrations, we can determine the temporal profile of cation concentration from the bases added. Using this method, we can measure a change in Co2+ concentration during a one hour period with an accuracy of 1 min. We also show the approach works for Zn2+ and Ca2+. We will present our methods for optimizing this biorecorder and characterize its performance in vitro. Recording data onto DNA with minutes time resolution could solve many challenging data acquisition problems in neuroscience and developmental biology, and could aid in the use of DNA as a data storage medium

Rosetta Brains: A Strategy for Molecularly-Annotated Connectomics

We propose a neural connectomics strategy called Fluorescent In-Situ Sequencing of Barcoded Individual Neuronal Connections (FISSEQ-BOINC), leveraging fluorescent in situ nucleic acid sequencing in fixed tissue (FISSEQ). FISSEQ-BOINC exhibits different properties from BOINC, which relies on bulk nucleic acid sequencing. FISSEQ-BOINC could become a scalable approach for mapping whole-mammalian-brain connectomes with rich molecular annotations

Enrichment of A Rare Subpopulation of miR-302-Expressing Glioma Cells by Serum Deprivation

Objective: MiR-302-367 is a cluster of polycistronic microRNAs that are exclusively expressed in embryonic stem (ES) cells. The miR-302-367 promoter is functional during embryonic development but is turned off in later stages. Motivated by the cancer stem cell hypothesis, we explored the potential expression of miR-302 in brain tumor cell lines. Materials and Methods: In the present experimental study, we have tried to expand our knowledge on the expression pattern and functionality of miR302 cluster by quantifying its expression in a series of glioma (A-172, 1321N1, U87MG) and medulloblastoma (DAOY) cell lines. To further assess the functionality of miR-302 in these cell lines, we cloned its promoter core region upstream of the enhanced green fluorescent protein (EGFP) or luciferase encoding genes. Results: Our data demonstrated a very low expression of miR-302 in glioma cell lines, compared with that of embryonal carcinoma cell line NT2 being used as a positive control. The expression of miR-302 promoter-EGFP construct in the aforementioned cell lines demonstrated GFP expression in a rare subpopulation of the cells. Serum deprivation led to the generation of tumorospheres, enrichment of miR-302 positive cells and upregulation of a number of pluripotency genes. Conclusion: Taken together, our data suggest that miR-302 could potentially be used as a novel putative cancer stem cell marker to identify and target cancer stem cells within tumor tissues

Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol

Purpose: The gene coding cytochrome P4501B1 (CYP1B1) has been shown to be a major cause of primary congenital glaucoma in the Iranian population. More recently it was shown to also be important in juvenile-onset open angle glaucoma (JOAG). We aimed to further investigate the role of CYP1B1 in a larger cohort of primary open angle glaucoma (POAG) patients which included late-onset patients. We also aimed to set up a microarray based protocol for mutation screening with an intent of using the protocol in a future population level screening program. Methods: Sixty three POAG patients, nine affected family members, and thirty three previously genotyped primary congenital glaucoma (PCG) patients were included in the study. Clinical examination included slit lamp biomicroscopy, IOP measurement, gonioscopic evaluation, fundus examination, and measurement of perimetry. G61E, R368H, R390H, and R469W were screened by a protocol that included multiplexed allele specific amplification in the presence of a protease (PrASE), use of sequence tagged primers, and hybridization to generic arrays on microarray slides. The entire coding sequences of CYP1B1 and myocilin (MYOC) genes were sequenced in all individuals assessed by the microarray assay to carry a mutation. Intragenic single nucleotide polymorphism (SNP) haplotpes were determined for mutated alleles. Results: Genotypes assessed by the array-based PrASE methodology were in 100 concordance with sequencing results. Seven mutation carrying POAG patients (11.1) were identified, and their distribution was quite skewed between the juvenile-onset individuals (5/21) as compared to late-onset cases (2/42). Four of the seven mutation carrying Iranian patients harbored two mutated alleles. CYP1B1 mutated alleles in Iranian PCG and POAG patients shared common haplotypes. MYOC mutations were not observed in any of the patients. Conclusions: The PrASE approach allowed reliable simultaneous genotyping of many individuals. It can be an appropriate tool for screening common mutations in large sample sizes. The results suggest that CYP1B1 is implicated in POAG among Iranians, notably in the juvenile-onset form. Contrary to POAG patients studied in other populations, many mutation harboring Iranian patients carry two mutated alleles. We propose an explanation for this observation. © 2008 Molecular Vision

Various indicators for the assessment of hospitals' performance status: Differences and similarities

Background: Hospitals are the most costly operational and really important units of health system because they consume about 50-89 of total health resources. Therefore efficient use of resources could help in saving and reallocating the financial and physical resources. Objectives: The aim of this study was to obtain an overview of hospitals' performance status by applying different techniques, to compare similarities and differences between these methods and suggest the most comprehensive and practical method of appraisal for managers and policy makers. Patients and Methods: This is a cross sectional study conducted in all hospitals of Ahvaz (eight hospitals affiliated with Jundishapur University of Medical Sciences and eight non-affiliated hospitals) during 2007 to 2011. Two kinds of data were collected through separate special checklists. Excel 2007 and Windeap 2.1 software were applied for data analysis. Results: The present findings show that the average of bed occupancy rate (BOR) in the studied hospitals was about 65.91 ± 1.16. The maximum number of inefficient hospitals in the present study happened in the years 2007, 2008 and 2010 (four hospitals) but there were two hospitals in the third part of the present graph which had maximum level of efficiency and optimal level of productivity in the years 2007 and 2009. Data Envelopment Analysis (DEA) showed that the mean score of technical efficiency for the studied hospitals is 0.924 ± 0.105 with the minimum of 0.585 ± 0.905 for hospital number 1. Furthermore It shows that only five hospitals (31.25) reach complete technical efficiency (TE) scores across all five years of 2007-11 (TE = 1). Conclusions: Results of the present and similar studies should be considered for the future planning and resource allocation of Iranian public hospitals. At the same time it is very important to consider need assessment results for each region according to its potentials, population under the coverage and other geographical and cultural indices. Furthermore because of potential limitations of each of the above models it is highly recommended to apply different methods of performance evaluation to reach a complete and real status view of the hospitals for future planning. © 2014, Iranian Red Crescent Medical Journal; Published by Kowsar Corp

Fluorescent in situ sequencing (FISSEQ) of RNA for gene expression profiling in intact cells and tissues

RNA sequencing measures the quantitative change in gene expression over the whole transcriptome, but it lacks spatial context. On the other hand, in situ hybridization provides the location of gene expression, but only for a small number of genes. Here we detail a protocol for genome-wide profiling of gene expression in situ in fixed cells and tissues, in which RNA is converted into cross-linked cDNA amplicons and sequenced manually on a confocal microscope. Unlike traditional RNA-seq our method enriches for context-specific transcripts over house-keeping and/or structural RNA, and it preserves the tissue architecture for RNA localization studies. Our protocol is written for researchers experienced in cell microscopy with minimal computing skills. Library construction and sequencing can be completed within 14 d, with image analysis requiring an additional 2 d