Molecular characterization of environmental non-Tuberculous mycobacteria using PCR- RFLP analysis of 441 Bp heat shock protein 65 fragments

Non- Tuberculous Mycobacteria are environmental opportunistic pathogens that can be found in various terrestrial and aquatic habitats. There are an epidemiological links between species isolated in tap water and those isolated from patients. hsp65 gene has more variability in its sequences, compared to the some more conserved genes in NTM, for identification of mycobacteria to species level. In this study, the prevalence of NTM in Isfahan City water samples was determined using culture, biochemical tests and PCR-RFLP analyses of hsp65 gene. Eighty-five water samples were collected and cultured. The mycobacterial isolates were identified by conventional biochemical tests. A 441 bp fragment of hsp65 genes was amplified and digested by two restriction enzymes, BstEII and HaeII. Digested products were analyzed using polyacrilamid gel electrophoresis [PAGE]. 25.9% of the water samples contained different species of NTM. Dominant isolates were M. fortuitum [26.7%], M. chelonae like organism [13.3%] and M. mucogenicum [13.3%]. Nineteen isolates of Mycobacteria were differentiated using hsp65 genes PCR-RFLP. Three isolates could not be identified at the species level because their RFLP patterns were different from other known PCR-RFLP profiles. There were different hsp65 gene PCR-RFLP profiles produced by digestion with BstEII and HaeIII. This study showed that PCR-RFLP of hsp65 gene in mycobacteria is more reliable method for identification of NTM at the specie level than conventional phenotypic methods [P<0.05]. In comparing of RFLP patterns of this study to other investigation, some minor differences were negligible

Construction of a new fusion protein vector associated to fibronectin binding protein A and clumping factor A derived from Staphylococcus aureus NCTC 8325

Staphylococcus aureus is a leading cause of many nosocomial and community acquired infections. According to many reports, antibiotic therapy cannot guarantee the eradication of S. aureus infections. Thus designing an adhesin based vaccine could restrain the S. aureus infections. This study designed for construction of a new fusion protein vaccine against S. aureus infections based on adhesin molecules fibronectin binding protein A [FnBPA] and clumping factor A [ClfA]. Bioinformatic experiments were performed using Oligo analyzer and DNAMAN softwares. The fragments corresponding to fnbA binding domain and a C-terminal fragment from clfA were amplified from S. aureus NCTC8325 genomic DNA. Purified PCR products and the vector, pET15b, were digested with NcoI and BamHI. The digested PCR products were hybridized together and then ligated to digested vector. Finally incomplete construct was assembled by Taq DNA polymerase. To quick confirmation of cloning procedure the new construct designated pfnbA-clfA was digested with NcoI and BamHI. To further verification, the product was sent for sequencing. The data based on bioinformatics analysis showed no homology between fusion protein and human proteins. Digestion of new vector with NcoI and BamHI confirmed the ligation of fusion protein sequence into pET15b. Sequencing results verified the integrity of target sequences. This study is the first effort to construct a new fusion protein vector based on S. aureus adhesins using a new design. This project is being continued to study the expression and biological activity of the fusion protein in a cell culture model

Prevalence of Porphyromonas gingivalis and Bacteroides forsythus in chronic periodontitis by multiplex PCR

Periodontitis is a common inflammatory and infectious disease which destroys the supporting structure of the teeth. Recent studies show that periodontal infection significantly increases the risk of some systemic diseases. It is generally accepted that bacterial species notably Porphyromonas gingivalis and Bacteroides fosythus are highly associated with periodontium. Molecular methods such as Multiplex PCR seem to be more sensitive and faster. Multiplex PCR alone can lower the limit of bacterial detection. Several pathogens can be detected simultaneously by this method. The Subgingival plaque samples from 61 patients including 34 women and 27 men in the age range of 24-69 years and an average age of 43 suffering from chronic periodontitis with probing depth of PD>/=6, and from 40 periodontally healthy controls including 22 women and 18 men in the age range of 21-69 years and an average age of 41.35 were collected by sterile curette. In this study, two species-specific forward primers were used in combination with a single reverse primer. The samples' DNA was extracted and Multiplex PCR was administered. Porphyromonas gingivalis was detected in 51 samples [81.61%] and 16 samples [40%] of the chronic periodontitis patients and the healthy subjects repectively. Moreover, Bacteroides forsythus was detected 32 samples [52.50%] of the chronic periodontitis patients but it was not detected in any of the samples from the healthy group. P. gingivalis and B. forsythus can be simultaneously detected using Multiplex PCR. The present data suggest that P. gingivalis is a more important factor in the etiology of chronic periodontitis. Further studies are needed to determine the spectrum of pathogenicity of the disease and effective management of diagnosis and treatment in order to decrease the risk of periodontal complications such as systemic infections

Job satisfaction among the staff in Al-Zahra training hospital

One of the managers' main concerns in maintaining and preserving human resources is developing job satisfaction. Job satisfaction enhances the staffs productivity and efficiency. In this study, staff's job satisfaction in one of the largest training hospitals was investigated. It was a cross - sectional study in which 142 people working in different wards were given a questionnaire. The questionnaire was used for the assessment of job satisfaction and its validity and reliability were checked in a pilot study. 69.8% of the staff had job satisfaction. They were satisfiedhighly [71.6%] with their department head, colleagues and their private life. They were satisfied the least [42.9%] with their income and what they had to do and 8% decided to leave the hospital because of unsatisfactory conditions. Scientific, purposeful and logical study of the staffs needs and punishing and rewarding systems and regarding the staffs views and suggestions to solve their current problems allow managers to develop job satisfaction and consequently promote human productivity