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Cloning, Expression, and Purification of Hyperthermophile α-Amylase from Pyrococcus woesei
Amir Ghasemi,Sobhan Ghafourian,Sedighe Vafaei,Reza Mohebi,Maryam Farzi,Morovat Taherikalani,Nourkhoda Sadeghifard 질병관리본부 2015 Osong Public Health and Research Persptectives Vol.6 No.6
Objectives: In an attempt α-amylase gene from Pyrococcus woesei was amplified and cloned into a pTYB2 vector to generate the recombinant plasmid pTY-α- amylase. Methods: Escherichia coli BL21 used as a host and protein expression was applied using IPTG. SDS-PAGE assay demonstrated the 100 kDa protein. Amylolytic activity of proteins produced by transformed E. coli cells was detected by zymography, and the rate of active α-amylase with and without the intein tag in both soluble conditions and as inclusion bodies solubilized by 4M urea were measured. Results: Amylolytic activity of ~185,000 U/L of bacterial culture was observed from the soluble form of the protein using this system. Conclusion: These results indicate that this expression system was appropriate for the production of thermostable α-amylase.
High Prevalence of AmpC β-Lactamases in Clinical Isolates of Escherichia coli in Ilam, Iran
Abbas Maleki,Afra Khosravi,Sobhan Ghafourian,Iraj Pakzad,Shiva Hosseini,Rashid Ramazanzadeh,Nourkhoda Sadeghifard 질병관리본부 2015 Osong Public Health and Research Persptectives Vol.6 No.3
Objectives: Widespread use of β-lactam antibiotics could cause resistance to this group of antibiotics in pathogenic bacteria through the production of the enzyme β-lactamases. The aimof this study is to determine themolecular detection of AmpC β-lactamases among clinical Escherichia coli isolated from Ilam hospitals in Ilam, Iran. Methods: One hundred and twelve clinical isolates of E. coli were collected from hospitalized patients and were identified by biochemical tests. They were evaluated for extended spectrum beta-lactamases (ESBLs) production, and the positive strains were subjected to AmpC enzymes; for detection of AmpC cluster genes, multiplex polymerase chain reaction was applied. Results: The analysis showed 62.5% of isolates were ESBLs positive and that five strains revealed the AmpC cluster genes. This is the first report of FOXM cluster genes in E. coli in Iran. Conclusion: Based on our results, the prevalence of AmpC β-lactamases is increasing in Iran, which caused failure in antibiotic therapy. So, the current study recommended the revision of antibiotic policy in Iranian hospitals.
Traditional and Modern Cell Culture in Virus Diagnosis
Ali Hematian,Nourkhoda Sadeghifard,Reza Mohebi,Morovat Taherikalani,Abbas Nasrolahi,Mansour Amraei,Sobhan Ghafourian 질병관리본부 2016 Osong Public Health and Research Persptectives Vol.7 No.2
Cell cultures are developed from tissue samples and then disaggregated by mechanical, chemical, and enzymatic methods to extract cells suitable for isolation of viruses. With the recent advances in technology, cell culture is considered a gold standard for virus isolation. This paper reviews the evolution of cell culture methods and demonstrates why cell culture is a preferred method for identification of viruses. In addition, the advantages and disadvantages of both traditional and modern cell culture methods for diagnosis of each type of virus are discussed. Detection of viruses by the novel cell culture methods is considered more accurate and sensitive. However, there is a need to include some more accurate methods such as molecular methods in cell culture for precise identification of viruses.
Anti-biofilm Properties of Peganum harmala against Candida albicans
Elham Aboualigalehdari,Nourkhoda Sadeghifard,Morovat Taherikalani,Zaynab Zargoush,Zahra Tahmasebi,Behzad Badakhsh,Arman Rostamzad,Sobhan Ghafourian,Iraj Pakzad 질병관리본부 2016 Osong Public Health and Research Persptectives Vol.7 No.2
Objectives: Vaginitis still remains as a health issue in women. It is notable that Candida albicans producing biofilm is considered a microorganism responsible for vaginitis with hard to treat. Also, Peganum harmala was applied as an anti fungal in treatment for many infections in Iran. Therefore, this study goal to investigate the role of P. harmala in inhibition of biofilm formation in C. albicans. Methods: So, 27 C. albicans collected from women with Vaginitis, then subjected for biofilm formation assay. P. harmala was applied as antibiofilm formation in C. albicans. Results: Our results demonstrated that P. harmala in concentration of 12 ㎍/ml easily inhibited strong biofilm formation; while the concentrations of 10 and 6 ㎍/ml inhibited biofilm formation in moderate and weak biofilm formation C. albicans strains, respectively. Conclusion: Hence, the current study presented P. harmala as antibiofilm herbal medicine for C. albicans; but in vivo study suggested to be performed to confirm its effectiveness.