REVIEW_ARTICLE Utilization of Site-Specific Recombination in Biopharmaceutical Production http://ibj.pasteur.ac.ir/article-1-1586-fa.pdf 2015-11-25 68 76 10.7508/ibj.2016.02.001 Utilization of Site-Specific Recombination in Biopharmaceutical Production Mammalian expression systems, due to their capacity in post-translational modification, are preferred systems for biopharmaceutical protein production. Several recombinant protein systems have been introduced to the market, most of which are under clinical development. In spite of significant improvements such as cell line engineering, introducing novel expression methods, gene silencing and process development, expression level is unpredictable and unstable because of the random location of integration in the genome. Site-specific recombination techniques are capable of producing stable and high producer clonal cells therefore, they are gaining more importance in the biopharmaceutical production. Site-specific recombination methods increase the recombinant protein production by specifically inserting a vector at a locus with specific expression trait. The present review focused on the latest developments in site-specific recombination techniques, their specific features and comparisons. http://ibj.pasteur.ac.ir/article-1-1586-en.pdf 2015-11-25 68 76 10.7508/ibj.2016.02.001 Site-specific recombinase Gene targeting CHO cell Maryam Ahmadi 1 AUTHOR Narges Damavandi 2 AUTHOR Mohammad Reza Akbari Eidgahi 3 AUTHOR Fatemeh Davami f_davami@pasteur.ac.ir 4 AUTHOR
ORIGINAL_ARTICLE Metabolomics-Based Study of Logarithmic and Stationary Phases of Promastigotes in Leishmania major by 1H NMR Spectroscopy http://ibj.pasteur.ac.ir/article-1-1581-fa.pdf 2015-11-23 77 83 10.7508/ibj.2016.02.002 Metabolomics-Based Study of Logarithmic and Stationary Phases of Promastigotes in Leishmania major by 1H NMR Spectroscopy Background: Cutaneous leishmaniasis is one of the most important parasitic diseases in humans. In this disease, one of the responsible organisms is Leishmania major, which is transmitted by sandfly vector. There are specific differences in biochemical profiles and metabolite pathways in logarithmic and stationary phases of Leishmania parasites. In the present study, 1H NMR spectroscopy was used to examine the metabolites outliers in the logarithmic and stationary phases of promastigotes in L. major to enlighten more about the transmission mechanism in metacyclogenesis of L. major. Methods: Promastigote was cultured, logarithmic and stationary phases were separated by the peanut agglutinin, and cell metabolites were extracted. 1H NMR spectroscopy was applied, and outliers were analyzed using principal component analysis. Results: The most altered metabolites in stationary and logarithmic phases were limited to citraconic acid, isopropylmalic acid, L-leucine, ornithine, caprylic acid, capric acid, and acetic acid. Conclusion: 1H NMR spectroscopy could play an important role in the characterization of metabolites in biochemical pathways during a metacyclogenesis process. These metabolites and their pathways can help in exploiting a transmission mechanism in metacyclogenesis, and outcoming data might be used in the metabolic network reconstruction of L. major modeling. http://ibj.pasteur.ac.ir/article-1-1581-en.pdf 2015-11-23 77 83 10.7508/ibj.2016.02.002 Leishmania major Metabolomics Principal component analysis Mohammad Arjmand 1 AUTHOR Azadeh Madrakian 2 AUTHOR Ghader Khalili 3 AUTHOR Ali Najafi Dastnaee 4 AUTHOR Zahra Zamani 5 AUTHOR Ziba Akbari Zibaakbari330@yahoo.com 6 AUTHOR
ORIGINAL_ARTICLE Identification of Sequence Variation in the Apolipoprotein A2 Gene and Their Relationship with Serum High-Density Lipoprotein Cholesterol Levels http://ibj.pasteur.ac.ir/article-1-1573-fa.pdf 2015-11-14 84 90 10.7508/ibj.2016.02.003 Identification of Sequence Variation in the Apolipoprotein A2 Gene and Their Relationship with Serum High-Density Lipoprotein Cholesterol Levels Background: Apolipoprotein A2 (APOA2) is the second major apolipoprotein of the high-density lipoprotein cholesterol (HDL-C). The study aim was to identify APOA2 gene variation in individuals within two extreme tails of HDL-C levels and its relationship with HDL-C level. Methods: This cross-sectional survey was conducted on participants from Tehran Glucose and Lipid Study (TLGS) at Research Institute for Endocrine Sciences, Tehran, Iran from April 2012 to February 2013. In total, 79 individuals with extreme low HDL-C levels (&le5th percentile for age and gender) and 63 individuals with extreme high HDL-C levels (&ge95th percentile for age and gender) were selected. Variants were identified using DNA amplification and direct sequencing. Results: Screen of all exons and the core promoter region of APOA2 gene identified nine single nucleotide substitutions and one microsatellite five of which were known and four were new variants. Of these nine variants, two were common tag single nucleotide polymorphisms (SNPs) and seven were rare SNPs. Both exonic substitutions were missense mutations and caused an amino acid change. There was a significant association between the new missense mutation (variant Chr.1:16119226, Ala98Pro) and HDL-C level. Conclusion: None of two common tag SNPs of rs6413453 and rs5082 contributes to the HDL-C trait in Iranian population, but a new missense mutation in APOA2 in our population has a significant association with HDL-C. http://ibj.pasteur.ac.ir/article-1-1573-en.pdf 2015-11-14 84 90 10.7508/ibj.2016.02.003 Apolipoprotein A-II Hyperlipidemia Polymorphism Mutation Genes Fatemeh Bandarian 1 AUTHOR Maryam Sadat Daneshpour 2 AUTHOR Mehdi Hedayati 3 AUTHOR Mohsen Naseri 4 AUTHOR Fereidoun Azizi Azizi@endocrine.ac.ir 5 AUTHOR
ORIGINAL_ARTICLE Comparison of Antimicrobial Susceptibility of Campylobacter Strains Isolated from Food Samples and Patients with Diarrhea http://ibj.pasteur.ac.ir/article-1-1585-fa.pdf 2015-11-25 91 96 10.7508/ibj.2016.02.004 Comparison of Antimicrobial Susceptibility of Campylobacter Strains Isolated from Food Samples and Patients with Diarrhea Background: Campylobacter infections may lead to serious conditions, including septicemia or other invasive forms of the disease, which require rapid and accurate laboratory diagnosis and subsequently appropriate antimicrobial therapy. The aim of this study was to compare the species distribution and antimicrobial susceptibility pattern of Campylobacter spp. strains isolated from patients and food samples. Methods: Biochemical identification was performed on 15 clinical and 30 food isolates of Campylobacter recovered onto Brucella agar containing 5% sheep blood. PCR was carried out to confirm the identity of Campylobacter spp. using primers for cadF, hipO, and asp genes of Campylobacter. To determine antibiotic sensitivity of isolates, Kirby-Bauer assay was carried out using 16 different antibiotic discs. Results: PCR assay and biochemical tests confirmed all 45 isolates as Campylobacter: 20 (44.44%) as C. jujeni, 10 (22.22%) as C. coli, and 15 (33.34%) as other Campylobacter strains. The maximum resistance was observed to cefotaxime and imipenem (each 86.49%) and the maximum sensitivity to erythromycin (48.65%). Conclusion: C. jujeni is dominant among isolates from clinical and food samples. In addition, tetracycline remains the first-line therapeutic agent against Campylobacter infections in Iran. http://ibj.pasteur.ac.ir/article-1-1585-en.pdf 2015-11-25 91 96 10.7508/ibj.2016.02.004 Campylobacter Genetic diversity Drug resistance Bita Bakhshi b.bakhshi@modares.ac.ir 1 AUTHOR Amin Naseri 2 AUTHOR Masoud Alebouyeh 3 AUTHOR
ORIGINAL_ARTICLE Chitosan-Based Intranasal Vaccine against Escherichia coli O157:H7 http://ibj.pasteur.ac.ir/article-1-1607-fa.pdf 2016-01-02 97 108 10.7508/ibj.2016.02.005 Chitosan-Based Intranasal Vaccine against Escherichia coli O157:H7 Background: Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is an infectious zoonotic pathogen causing human infections. These infections, in some cases, can lead to hemolytic uremic syndrome and its life-threatening complications and even death worldwide. The first intimate bacterial adhesion, intimin (I), with its own receptor translocated intimin receptor (Tir) and E. coli secreted protein A, acting as Tir conduit, are highly immunogenic proteins for vaccine development against E. coli O157:H7. Methods: A chimeric trivalent recombinant protein was previously found to be a suitable strategy for developing vaccines against E. coli O157:H7. In this study, the recombinant EIT (rEIT) was used to design a protective EHEC nasal nanovaccine. Chitosan and its water-soluble derivative, trimethylated chitosan (TMC), as muco-adhesive biopolymers, are good candidates for preparation of nanovaccines.  Using the electrospraying technique, as a novel method, we could obtain particles of rEIT loaded with chitosan and TMC on a nanometer scale. Mice were immunized with intranasal administration or intrapretoneal injection of rEIT. Results: The rEIT-specific immune responses (IgG and IgA) were measured by indirect ELISA. Only nasal administration of chitosan electrospray and TMC formulation produced significant secretion IgA. Intranasal administration of nanovaccine reduced the duration of bacterial fecal shedding on mice challenged with E. coli O157:H7. Conclusion: Since development of mucosal vaccines for the prevention of infectious diseases requires efficient antigen delivery therefore, this research could be a new strategy for developing vaccine against E. coli O157:H7. http://ibj.pasteur.ac.ir/article-1-1607-en.pdf 2016-01-02 97 108 10.7508/ibj.2016.02.005 Enterohemorrhagic Escherichia coli Nanoparticles Intranasal vaccination Tahere Doavi 1 AUTHOR Seyed Latif Mousavi slmousavi@shahed.ac.ir 2 AUTHOR Mehdi Kamali 3 AUTHOR Jafar Amani 4 AUTHOR Mahdi Fasihi Ramandi 5 AUTHOR
ORIGINAL_ARTICLE Characteristics of Human Endometrial Stem Cells in Tissue and Isolated Cultured Cells: An Immunohistochemical Aspect http://ibj.pasteur.ac.ir/article-1-1575-fa.pdf 2015-11-14 109 116 10.7508/ibj.2016.02.006 Characteristics of Human Endometrial Stem Cells in Tissue and Isolated Cultured Cells: An Immunohistochemical Aspect Background: The aim of this study was to investigate the percentage of the stem cells population in human endometrial tissue sections and cultured cells at fourth passage. Methods: Human endometrial specimens were divided into two parts, one part for morphological studies and the other part for in vitro culture. Full thickness of human normal endometrial sections and cultured endometrial cells at fourth passage were analyzed via immunohistochemistry for CD146 and some stemness markers such as Oct4, Nanog, Sox2, and Klf4 and the expression of typical mesenchymal stem cell markers CD90, CD105. Results: 11.88 ± 1.29% of human endometrial cells whitin tissue sections expressed CD146 marker vs. 28±2.3% of cultured cells, CD90 and CD105 were expressed by functionalis stroma (85±2.4 and 89±3.2%) than basalis stroma (16±1.4 and 17±1.9%), respectively (P<0.05). Oct4 and Nanog-expressing cells comprise 1.43±0.08 and 0.54±0.01% of endometrial stromal cells in endometrial sections vs. 12±3.1% and 8±2.9% of cultured cells, respectively. They reside near the glands in the basal layer of endometrium. Sox2 and Klf4 were not commonly expressed in tissue samples and cultured cells. CD9 and EpCAM were expressed by epithelial cells of the endometrium, rather than by stroma or perivascular cells. Conclusion: The human endometrial stem cells and pluripotency markers may be localized more in basalis layer of endometrium. The immunostaining observations of endometrial cells at fourth passage were correlated with the immunohistochemistry data. http://ibj.pasteur.ac.ir/article-1-1575-en.pdf 2015-11-14 109 116 10.7508/ibj.2016.02.006 Endometrium Immunohistochemistry Mesenchymal stem cells Mehri Fayazi 1 AUTHOR Mojdeh Salehnia mogdeh@dr.com 2 AUTHOR Saeideh Ziae 3 AUTHOR
ORIGINAL_ARTICLE Investigation of Histone Lysine-Specific Demethylase 5D (KDM5D) Isoform Expression in Prostate Cancer Cell Lines: a System Approach Background: It is now well-demonstrated that histone demethylases play an important role in developmental controls, cell-fate decisions, and a variety of diseases such as cancer. Lysine-specific demethylase 5D (KDM5D) is a male-specific histone demethylase that specifically demethylates di- and tri-methyl H3K4 at the start site of active genes. In this light, the aim of this study was to investigate isoform/transcript-specific expression profiles of KDM5D in three prostate cancer cell lines, Du-145, LNCaP, and PC3. Methods: Real-time PCR analysis was performed to determine the expression levels of different KDM5D transcripts in the prostate cell lines. A gene regulatory network was established to analyze the gene expression profile. Results: Significantly different expression levels of both isoforms were found among the three cell lines. Interestingly, isoform I was expressed in three cell lines while isoform III did only in DU-145. The expression levels of both isoforms were higher in DU-145 when compared to other cell lines (P<0.0001). The observed expression profile was determined by using regulatory network analyses. Conclusion: The present study, for the first time, not only showed the expression profiles of KDM5D isoforms in prostate cancer cell lines but also evaluated the effects of the gene regulatory network on the expression profile of this gene. http://ibj.pasteur.ac.ir/article-1-1608-en.pdf 2016-01-03 117 121 10.7508/ibj.2016.02.007 KDM5D Prostate cancer Regulatory networks Zohreh Jangravi 1 AUTHOR Mohammad Najafi 2 AUTHOR Mohammd Shabani shabani.mo@iums.ac.ir 3 AUTHOR
SHORT_COMMUNICATION Association Study of rs1333040 and rs1004638 Polymorphisms in the 9p21 Locus with Coronary Artery Disease in Southwest of Iran http://ibj.pasteur.ac.ir/article-1-1693-fa.pdf 2016-04-27 122 127 10.7508/ibj.2016.02.008 Association Study of rs1333040 and rs1004638 Polymorphisms in the 9p21 Locus with Coronary Artery Disease in Southwest of Iran Background: Coronary artery disease (CAD) is a multifactorial and heterogenic disease. Recently, genome-wide association studies have reported that rs1333040 (C/T) and rs1004638 (A/T) single nucleotide polymorphisms (SNPs) in the 9p21 locus have very strong association with CAD. This study aimed to examine these associations in Southwest of Iran. Methods: Blood samples were collected from 200 CAD patients and 110 healthy individuals with no CAD. The association of two SNPs with CAD was evaluated by PCR and restriction fragment length polymorphism. Results: Chi-square test showed no association between rs1333040 SNP and CAD (X2: 4.66, df: 2, P=0.09). Also, there was no association between rs1004638 SNP and CAD (X2: 0.27, df: 2, P=0.88). Conclusion: No association was observed between rs1333040 and rs1004638 SNPs in the 9P21 region and CAD in Southwest of Iran. http://ibj.pasteur.ac.ir/article-1-1693-en.pdf 2016-04-27 122 127 10.7508/ibj.2016.02.008 Coronary artery disease Single nucleotide polymorphisms Genetic association study Iran Khadijeh Golabgir Khademi 1 AUTHOR Ali Mohammad Foroughmand Foroughmand.a@scu.ac.ir 2 AUTHOR Hamid Galehdari 3 AUTHOR Saied Yazdankhah 4 AUTHOR Mahdi Pourmahdi Borujeni 5 AUTHOR Zahra Shahbazi 6 AUTHOR Parvaneh Dinarvand 7 AUTHOR