Introduction: The anti-infective effect of early colonization of infants by potentially probiotic lactic acid bacteria in human milk is a growing area of research. Lactobacillus plantarum colonization in early infancy may be important to health in later life. Here, we present an investigation into the presence of L. plantarum in breast milk from Iranian mothers. Materials and Methods: Human breast milk samples (n = 40) were randomly collected from lactating and breastfeeding women having undergone full-term pregnancies. Information concerning personal characteristics was collected after birth. The samples were cultured in de Man, Rogosa, and Sharpe medium using the pour plate technique, and isolates were initially identified by biochemical methods. Isolates were established as belonging to the genus Lactobacillus based on the 16- 23S rRNA region, and the species L. plantarum was identified using PCR and primers targeting the recA gene. Results: In our study, 35 samples (87.5%) contained suspected lactobacilli based on phenotypic tests. Thirty of these (85.71%) were confirmed as containing bacteria of the genus Lactobacillus using a genotypic method (PCR), all of which were found to be L. plantarum. Conclusion: Probiotic bacteria in a mother’s breast milk may have positive effects on her infant’s health. This insight creates new perspectives concerning the use of breast milk as a source of probiotic bacteria for bacteriotherapy.
{"title":"Identification of Lactobacillus plantarum in Breast Milk","authors":"Mansoureh Taghizadeh, H. Safaei, F. Poursina","doi":"10.18502/RMM.V5I4.3065","DOIUrl":"https://doi.org/10.18502/RMM.V5I4.3065","url":null,"abstract":"Introduction: The anti-infective effect of early colonization of infants by potentially probiotic lactic acid bacteria in human milk is a growing area of research. Lactobacillus plantarum colonization in early infancy may be important to health in later life. Here, we present an investigation into the presence of L. plantarum in breast milk from Iranian mothers. Materials and Methods: Human breast milk samples (n = 40) were randomly collected from lactating and breastfeeding women having undergone full-term pregnancies. Information concerning personal characteristics was collected after birth. The samples were cultured in de Man, Rogosa, and Sharpe medium using the pour plate technique, and isolates were initially identified by biochemical methods. Isolates were established as belonging to the genus Lactobacillus based on the 16- 23S rRNA region, and the species L. plantarum was identified using PCR and primers targeting the recA gene. Results: In our study, 35 samples (87.5%) contained suspected lactobacilli based on phenotypic tests. Thirty of these (85.71%) were confirmed as containing bacteria of the genus Lactobacillus using a genotypic method (PCR), all of which were found to be L. plantarum. Conclusion: Probiotic bacteria in a mother’s breast milk may have positive effects on her infant’s health. This insight creates new perspectives concerning the use of breast milk as a source of probiotic bacteria for bacteriotherapy. ","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42538240","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mehrnaz Bakhti, M. Haghshenas, R. Valadan, Mehdi Rabie Rudsari
Introduction: Human immunodeficiency virus (HIV) infection increases the risk of infection with other pathogens, including hepatitis B virus (HBV) and hepatitis C virus (HCV). A crucial aspect of HIV prevention and treatment programs is knowledge of the prevalence of co-infection of HIV and HBV and/or HCV. This study sought to determine HBV and HCV co-infection in HIV-positive patients in northern Iran. Materials and Methods: Blood samples were collected from 83 HIV-positive patients whose infection was previously confirmed by real-time polymerase chain reaction in the HIV center in the North of Iran. A structured questionnaire was used to obtain socio-demographic data from participants. Samples were screened for hepatitis B surface antigen and anti-HCV antibody. All non-reactive samples were recorded as negative. Results: The 83 patients comprised 50 (60%) males and 33 (40%) females. Twenty eight (33%) and 15 (18%) subjects were positive for HCV antibody and hepatitis B surface antigen, respectively. Seven (8%) of subjects were co-infected with all three viruses. Conclusion: Seroprevalence of HCV and HIV co-infection was high and was strongly related to mutual acquisition.
{"title":"Prevalence of HBV/HCV Infections in HIV-Positive Patients in Northern Iran","authors":"Mehrnaz Bakhti, M. Haghshenas, R. Valadan, Mehdi Rabie Rudsari","doi":"10.18502/RMM.V5I4.3066","DOIUrl":"https://doi.org/10.18502/RMM.V5I4.3066","url":null,"abstract":"Introduction: Human immunodeficiency virus (HIV) infection increases the risk of infection with other pathogens, including hepatitis B virus (HBV) and hepatitis C virus (HCV). A crucial aspect of HIV prevention and treatment programs is knowledge of the prevalence of co-infection of HIV and HBV and/or HCV. This study sought to determine HBV and HCV co-infection in HIV-positive patients in northern Iran. Materials and Methods: Blood samples were collected from 83 HIV-positive patients whose infection was previously confirmed by real-time polymerase chain reaction in the HIV center in the North of Iran. A structured questionnaire was used to obtain socio-demographic data from participants. Samples were screened for hepatitis B surface antigen and anti-HCV antibody. All non-reactive samples were recorded as negative. Results: The 83 patients comprised 50 (60%) males and 33 (40%) females. Twenty eight (33%) and 15 (18%) subjects were positive for HCV antibody and hepatitis B surface antigen, respectively. Seven (8%) of subjects were co-infected with all three viruses. Conclusion: Seroprevalence of HCV and HIV co-infection was high and was strongly related to mutual acquisition. ","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46030772","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Maryam Samareh Salavati Pour, Fatemeh Hoseinpour Kasgari, A. Farsinejad, A. Fatemi, R. M. Khalilabadi
Introduction: Mesenchymal stem cells (MSCs) are widely studied due to their self- renewal potential and capacity to differentiate into multiple tissues. However, they have a limited life span of several divisions in vitro, which alters various cellular characteristics and reduces their application. Aim: We evaluated the effect of platelet-derived microparticles on gene expression of hTERT, one of the main factors involved in aging and cell longevity. Materials and methods: Umbilical cord MSCs were used for this study. Cells were characterized by evaluating morphology via inverted microscope and identifying associated surface markers using flow cytometry. Platelet-derived microparticles were prepared by centrifuging platelet bags at varying speeds, and their concen- trations were determined by Bradford assay. At 30% confluency, MSCs were treated with 50 μg/mL of microparticles for five days. Then, RNA was extracted and cDNA was synthesized. Quantitative expression of hTERT was assessed using real-time polymerase chain reaction (PCR). Results: Fibroblast-like cells were isolated from umbilical cord tissue and MSCs were identified by the presence of mesenchymal surface markers via flow cytometry. Real- time PCR showed that gene expression of hTERT increased by more than three times when treated with platelet-derived microparticles, in comparison to expression of the control group. Conclusion: We concluded that platelet-derived microparticles may be a potentially safe and effective method to increase hTERT gene expression in MSCs, ultimately prolonging their life span in vitro.
{"title":"Platelet-Derived Microparticles Increase Expression of hTERT in Umbilical Cord Mesenchymal Stem Cells","authors":"Maryam Samareh Salavati Pour, Fatemeh Hoseinpour Kasgari, A. Farsinejad, A. Fatemi, R. M. Khalilabadi","doi":"10.18502/RMM.V5I4.3063","DOIUrl":"https://doi.org/10.18502/RMM.V5I4.3063","url":null,"abstract":"Introduction: Mesenchymal stem cells (MSCs) are widely studied due to their self- renewal potential and capacity to differentiate into multiple tissues. However, they have a limited life span of several divisions in vitro, which alters various cellular characteristics and reduces their application. Aim: We evaluated the effect of platelet-derived microparticles on gene expression of hTERT, one of the main factors involved in aging and cell longevity. Materials and methods: Umbilical cord MSCs were used for this study. Cells were characterized by evaluating morphology via inverted microscope and identifying associated surface markers using flow cytometry. Platelet-derived microparticles were prepared by centrifuging platelet bags at varying speeds, and their concen- trations were determined by Bradford assay. At 30% confluency, MSCs were treated with 50 μg/mL of microparticles for five days. Then, RNA was extracted and cDNA was synthesized. Quantitative expression of hTERT was assessed using real-time polymerase chain reaction (PCR). Results: Fibroblast-like cells were isolated from umbilical cord tissue and MSCs were identified by the presence of mesenchymal surface markers via flow cytometry. Real- time PCR showed that gene expression of hTERT increased by more than three times when treated with platelet-derived microparticles, in comparison to expression of the control group. Conclusion: We concluded that platelet-derived microparticles may be a potentially safe and effective method to increase hTERT gene expression in MSCs, ultimately prolonging their life span in vitro. ","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48170332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Fatemeh Hoseinpour Kasgari, Maryam Samareh Salavati Pour, A. Farsinejad, A. Fatemi, R. M. Khalilabadi
ntroduction: Due to high differentiation potential and self-renewality, Mesenchymal stem cells are now widely considered by researchers in several diseases. FBS is used as a supplement in culture media for proliferation, differentiation, and other culture processes of MSCs, which is associated with transmission risk of a variety of infections as well as immune responses. PRGF derived from platelets contains growth factors causing the growth and proliferation of MSCs. This study was conducted to compare the effect of PRGF in comparison to FBS on the expression of h-TERT gene, in umbilical cord-derived MSCs. Materials and Methods: This study is an experimental research. Four expired platelet concentrate bags were obtained from Kerman blood transfusion center, and PRGF was prepared by multiple centrifugation rounds of the platelet bag. Calcium chloride was added as an anticoagulant to PRGF in order to prevent gelatinization of the culture medium. On the other hand, mesenchymal stem cells were isolated from the umbilical cord as a primary culture. The phenotype of the cells was confirmed by flow cytometry, and the cells were randomly cultured as control (using FBS) and experimental (using PRGF) groups. The expression of the gene involved in increasing cell longevity (h-TERT) was investigated by real-time PCR technique after six days. Results: Mesenchymal stem cells were successfully isolated from the umbilical cord. Morphologically, the mesenchymal cells cultured in the experimental group (using PRGF) were similar to the cells in the control medium. The cells exhibited a high expression level of CD73, CD90, and CD105, while the surface markers of hematopoietic cells such as CD45 and CD34 were slightly expressed. Therefore, there was no significant difference in the expression of cell surface markers between control and experimental groups. In this study, using the real-time PCR technique, it was shown that PRGF derived from the platelet could increase the expression of h- TERT gene in the umbilical cord mesenchymal stem cells compared with the control. (P = 0.034)
{"title":"Platelet Rich in Growth Factors (PRGF): A Suitable Replacement for Fetal Bovine Serum (FBS) in Mesenchymal Stem Cell Culture","authors":"Fatemeh Hoseinpour Kasgari, Maryam Samareh Salavati Pour, A. Farsinejad, A. Fatemi, R. M. Khalilabadi","doi":"10.18502/RMM.V5I4.3061","DOIUrl":"https://doi.org/10.18502/RMM.V5I4.3061","url":null,"abstract":" ntroduction: Due to high differentiation potential and self-renewality, Mesenchymal stem cells are now widely considered by researchers in several diseases. FBS is used as a supplement in culture media for proliferation, differentiation, and other culture processes of MSCs, which is associated with transmission risk of a variety of infections as well as immune responses. PRGF derived from platelets contains growth factors causing the growth and proliferation of MSCs. This study was conducted to compare the effect of PRGF in comparison to FBS on the expression of h-TERT gene, in umbilical cord-derived MSCs. Materials and Methods: This study is an experimental research. Four expired platelet concentrate bags were obtained from Kerman blood transfusion center, and PRGF was prepared by multiple centrifugation rounds of the platelet bag. Calcium chloride was added as an anticoagulant to PRGF in order to prevent gelatinization of the culture medium. On the other hand, mesenchymal stem cells were isolated from the umbilical cord as a primary culture. The phenotype of the cells was confirmed by flow cytometry, and the cells were randomly cultured as control (using FBS) and experimental (using PRGF) groups. The expression of the gene involved in increasing cell longevity (h-TERT) was investigated by real-time PCR technique after six days. Results: Mesenchymal stem cells were successfully isolated from the umbilical cord. Morphologically, the mesenchymal cells cultured in the experimental group (using PRGF) were similar to the cells in the control medium. The cells exhibited a high expression level of CD73, CD90, and CD105, while the surface markers of hematopoietic cells such as CD45 and CD34 were slightly expressed. Therefore, there was no significant difference in the expression of cell surface markers between control and experimental groups. In this study, using the real-time PCR technique, it was shown that PRGF derived from the platelet could increase the expression of h- TERT gene in the umbilical cord mesenchymal stem cells compared with the control. (P = 0.034) ","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41450847","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Meysam Aghajani, A. Rafiei, J. Ghaffari, R. Valadan, M. Kardan
.
{"title":"Immune Dysregulation in Children with Allergic Asthma: A Close Relationship Between IL-17 but not IL-4 or IFN-","authors":"Meysam Aghajani, A. Rafiei, J. Ghaffari, R. Valadan, M. Kardan","doi":"10.18502/RMM.V6I1.3926","DOIUrl":"https://doi.org/10.18502/RMM.V6I1.3926","url":null,"abstract":"<jats:p>.</jats:p>","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49446642","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
R. Vinodhini, Legesse Kebede, G. Teka, Bersisa Asana, T. Abel
1 Unit of Biochemistry, Department of Medicine, College of Medicine and Health Sciences, Ambo University, Ambo Town, Ethiopia. 2 Department of Internal Medicine, Ambo University Referral Hospital, Ambo Town, Ethiopia. 3 Department of Public Health, College of Medicine and Health Sciences, Ambo University, Ambo Town, Ethiopia. 4 Department of Surgery, Ambo University Referral Hospital, Ambo Town, Ethiopia. 5 Department of Surgery, Aka-Kotebe General Hospital, Addis Ababa, Ethiopia.
{"title":"Prevalence of Prediabetes and its Risk Factors among the Employees of Ambo University, Oromia Region, Ethiopia","authors":"R. Vinodhini, Legesse Kebede, G. Teka, Bersisa Asana, T. Abel","doi":"10.29252/RMM.5.3.11","DOIUrl":"https://doi.org/10.29252/RMM.5.3.11","url":null,"abstract":"1 Unit of Biochemistry, Department of Medicine, College of Medicine and Health Sciences, Ambo University, Ambo Town, Ethiopia. 2 Department of Internal Medicine, Ambo University Referral Hospital, Ambo Town, Ethiopia. 3 Department of Public Health, College of Medicine and Health Sciences, Ambo University, Ambo Town, Ethiopia. 4 Department of Surgery, Ambo University Referral Hospital, Ambo Town, Ethiopia. 5 Department of Surgery, Aka-Kotebe General Hospital, Addis Ababa, Ethiopia.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"5 1","pages":"11-20"},"PeriodicalIF":0.0,"publicationDate":"2017-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44454054","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Mona Akhondnezhad, Mehrnaz Bakhti, M. Nasrolahei, B. Shabankhani, H. Goli
Corresponding Author: Hamid Reza Goli Department of Microbiology, Faculty of Medicine, KM 18 Khazarabad Road, Khazar Sq, Sari, Iran. Phone: +98-1133542067 E-mail: goli59@gmail.com Abstract Background: Enterococci are important gram-positive bacteria causing dental calculus in human beings; however, the role of these bacteria in oral cavity is unclear. The aim of this study was to investigate the presence of Enterococcal Surface Protein (esp) gene in Enterococcus faecalis isolated from dental calculus in the city of Sari, Iran. Materials and Methods: In the present study, 207 dental calculus samples were collected from patients. The isolates were identified by growth on Bile Esculin agar, Gram stain, Catalase test, Growth at 6.5% NaCl, PYR and arabinose fermentation test. Antimicrobial susceptibility pattern of the isolates was determined by disk agar diffusion method. The presence of esp gene was assessed by polymerase chain reaction (PCR). Results: Among the 56 (27%) enterococci isolated from dental calculus, 43 (76.7%) were determined as E. faecalis. The resistance rate to ampicillin, vancomycin, tetracycline, ciprofloxacin and erythromycin in E. faecalis isolates was estimated as 13.9%, 4.6%, 11.6%, 6.9% and 13.9%, respectively. The esp gene was detected in 18.6% of E. faecalis isolates. Among the isolates containing esp gene, 33.3%, 50%, 40%, 33.3% and 33.3% of them were resistant to ampicillin, vancomycin, tetracycline, ciprofloxacin and erythromycin, respectively. Conclusion: E. faecalis is an important organism causing dental calculus but the presence of esp gene had no correlation with the resistance to tested antimicrobial agents.
{"title":"Molecular Detection of Enterococcal Surface Protein (esp) Gene in Enterococcus faecalis Isolated from Dental Calculus of Patients in Sari, Iran","authors":"Mona Akhondnezhad, Mehrnaz Bakhti, M. Nasrolahei, B. Shabankhani, H. Goli","doi":"10.29252/RMM.5.3.21","DOIUrl":"https://doi.org/10.29252/RMM.5.3.21","url":null,"abstract":"Corresponding Author: Hamid Reza Goli Department of Microbiology, Faculty of Medicine, KM 18 Khazarabad Road, Khazar Sq, Sari, Iran. Phone: +98-1133542067 E-mail: goli59@gmail.com Abstract Background: Enterococci are important gram-positive bacteria causing dental calculus in human beings; however, the role of these bacteria in oral cavity is unclear. The aim of this study was to investigate the presence of Enterococcal Surface Protein (esp) gene in Enterococcus faecalis isolated from dental calculus in the city of Sari, Iran. Materials and Methods: In the present study, 207 dental calculus samples were collected from patients. The isolates were identified by growth on Bile Esculin agar, Gram stain, Catalase test, Growth at 6.5% NaCl, PYR and arabinose fermentation test. Antimicrobial susceptibility pattern of the isolates was determined by disk agar diffusion method. The presence of esp gene was assessed by polymerase chain reaction (PCR). Results: Among the 56 (27%) enterococci isolated from dental calculus, 43 (76.7%) were determined as E. faecalis. The resistance rate to ampicillin, vancomycin, tetracycline, ciprofloxacin and erythromycin in E. faecalis isolates was estimated as 13.9%, 4.6%, 11.6%, 6.9% and 13.9%, respectively. The esp gene was detected in 18.6% of E. faecalis isolates. Among the isolates containing esp gene, 33.3%, 50%, 40%, 33.3% and 33.3% of them were resistant to ampicillin, vancomycin, tetracycline, ciprofloxacin and erythromycin, respectively. Conclusion: E. faecalis is an important organism causing dental calculus but the presence of esp gene had no correlation with the resistance to tested antimicrobial agents.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"5 1","pages":"21-25"},"PeriodicalIF":0.0,"publicationDate":"2017-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45805636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sanaz Yari, F. Behzadian, H. R. Nejad, M. Masoumian, M. Karimi
Corresponding Author: Farida Behzadian Department of Biosciences and Biotechnology, Malek-Ashtar University of Technology, Tehran, Iran. Phone: +98-2122974603 E-mail: fbehzadian@yahoo.com Abstract Background: Parathyroid Hormone (PTH) is secreted by parathyroid glands and controls the level of calcium in bones and kidney. PTH is a small polypeptide with 84 amino acids, but the first 34 amino acids of which are enough for hormone biological activity and can be used in the treatment of Osteoporosis. The expression efficiency of recombinant human parathyroid hormone rhPTH (1-34) or Teriparatide using a cleavable fusion protein strategy was compared in two strains of E. coli. Materials and Methods: A cassette was designed and fully synthesized for prokaryotic expression of rhPTH using pET system. From 5’ to 3’, the cassette consisted of: Trx tag to increase the solubility of protein, His tag for purification and detection of protein, enterokinase site to cleave all fusion moieties, and an optimized gene code for Teriparatide corresponding to the amino acid sequence of hPTH. This cassette was cloned into pET32a vector. The vector was simultaneously transformed and expressed in two different E. coli strains. The ability of strains for expression of this recombinant pharmaceutical was compared. Early expression was confirmed by SDS-PAGE and Western Blotting. The soluble fusion protein was harvested and purified by immobilized affinity chromatography. Then the fusion moiety was released from Teriparatide by enterokinase digestion. Results: The fusion form of rhPTH was efficiently expressed in both E. coli strains. However, the percentage of the target protein to the total protein content in Rosetta-gami was more than its amount in BL21 (60 % vs 25%).The fusion protein was highly purified with Ni-NTA column. Up to 18.5 mg/ml of pure fusion protein has been obtained from 1-liter Rosetta-gami strain of E. coli. The pure Teriparatide was released by enterokinase digestion. Conclusion: The pure rhPTH (1-34) produced here, could be the subject for biological activity and quality control assessments, and following formulation processing, it could be applied as a peptide drug in the treatment of Osteoporosis.
{"title":"Expression and Purification of Soluble form of Human Parathyroid Hormone (rhPTH1-34) by Trx Tag in E. coli","authors":"Sanaz Yari, F. Behzadian, H. R. Nejad, M. Masoumian, M. Karimi","doi":"10.29252/RMM.5.3.26","DOIUrl":"https://doi.org/10.29252/RMM.5.3.26","url":null,"abstract":"Corresponding Author: Farida Behzadian Department of Biosciences and Biotechnology, Malek-Ashtar University of Technology, Tehran, Iran. Phone: +98-2122974603 E-mail: fbehzadian@yahoo.com Abstract Background: Parathyroid Hormone (PTH) is secreted by parathyroid glands and controls the level of calcium in bones and kidney. PTH is a small polypeptide with 84 amino acids, but the first 34 amino acids of which are enough for hormone biological activity and can be used in the treatment of Osteoporosis. The expression efficiency of recombinant human parathyroid hormone rhPTH (1-34) or Teriparatide using a cleavable fusion protein strategy was compared in two strains of E. coli. Materials and Methods: A cassette was designed and fully synthesized for prokaryotic expression of rhPTH using pET system. From 5’ to 3’, the cassette consisted of: Trx tag to increase the solubility of protein, His tag for purification and detection of protein, enterokinase site to cleave all fusion moieties, and an optimized gene code for Teriparatide corresponding to the amino acid sequence of hPTH. This cassette was cloned into pET32a vector. The vector was simultaneously transformed and expressed in two different E. coli strains. The ability of strains for expression of this recombinant pharmaceutical was compared. Early expression was confirmed by SDS-PAGE and Western Blotting. The soluble fusion protein was harvested and purified by immobilized affinity chromatography. Then the fusion moiety was released from Teriparatide by enterokinase digestion. Results: The fusion form of rhPTH was efficiently expressed in both E. coli strains. However, the percentage of the target protein to the total protein content in Rosetta-gami was more than its amount in BL21 (60 % vs 25%).The fusion protein was highly purified with Ni-NTA column. Up to 18.5 mg/ml of pure fusion protein has been obtained from 1-liter Rosetta-gami strain of E. coli. The pure Teriparatide was released by enterokinase digestion. Conclusion: The pure rhPTH (1-34) produced here, could be the subject for biological activity and quality control assessments, and following formulation processing, it could be applied as a peptide drug in the treatment of Osteoporosis.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"5 1","pages":"26-31"},"PeriodicalIF":0.0,"publicationDate":"2017-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46445300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Corresponding Author: Hassan Habibi Assistant professor of Agriculture and natural sciences college, Persian Gulf University, Bushehr, Iran. Phone: +98-9173034921 E-mail: H.habibi@pgu.ac.ir Abstract Background: Researchers are seeking new plant compounds as an alternative to chemical drugs and antibiotics due to the increasing resistance of pathogenic bacteria to antibiotics. This study investigated the antibacterial effect of Plantago ovata and Lallemantia iberica L. seed extracts on some foodborne human pathogenic bacteria. Materials and Methods: Disk-diffusion antibiotic sensitivity testing, Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were used to evaluate the antibacterial activity of plant extracts in comparison to the tetracycline, as a control antibiotic. Results: The results of this experiment showed that the L. iberica seed extract had the greatest effect on Bacillus subtilis, Bacillus sphaericus and Pseudomonas aeruginosa and did not have inhibitory effect or moderate inhibitory effect against other bacteria. Also, P. ovata extract had a high and moderate effect against Bacillus sphaericus and Pseudomonas aeruginosa, respectively. This extract had no inhibitory effect on the other bacteria. Tetracycline also had a significant inhibitory effect on all tested bacteria. Conclusion: According to the results of this study, it can be concluded that extracts of some Iranian native plants can be a suitable alternative to the existing antibiotics.
{"title":"Antibacterial Effect of Plantago Ovata and Lallemantia Iberica Seed Extracts against Some Bacteria","authors":"L. Karami, N. Ghahtan, H. Habibi","doi":"10.29252/RMM.5.3.32","DOIUrl":"https://doi.org/10.29252/RMM.5.3.32","url":null,"abstract":"Corresponding Author: Hassan Habibi Assistant professor of Agriculture and natural sciences college, Persian Gulf University, Bushehr, Iran. Phone: +98-9173034921 E-mail: H.habibi@pgu.ac.ir Abstract Background: Researchers are seeking new plant compounds as an alternative to chemical drugs and antibiotics due to the increasing resistance of pathogenic bacteria to antibiotics. This study investigated the antibacterial effect of Plantago ovata and Lallemantia iberica L. seed extracts on some foodborne human pathogenic bacteria. Materials and Methods: Disk-diffusion antibiotic sensitivity testing, Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were used to evaluate the antibacterial activity of plant extracts in comparison to the tetracycline, as a control antibiotic. Results: The results of this experiment showed that the L. iberica seed extract had the greatest effect on Bacillus subtilis, Bacillus sphaericus and Pseudomonas aeruginosa and did not have inhibitory effect or moderate inhibitory effect against other bacteria. Also, P. ovata extract had a high and moderate effect against Bacillus sphaericus and Pseudomonas aeruginosa, respectively. This extract had no inhibitory effect on the other bacteria. Tetracycline also had a significant inhibitory effect on all tested bacteria. Conclusion: According to the results of this study, it can be concluded that extracts of some Iranian native plants can be a suitable alternative to the existing antibiotics.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"5 1","pages":"32-36"},"PeriodicalIF":0.0,"publicationDate":"2017-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49196734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Corresponding Author: Amin Talebi Bezmin Abadi Department of Bacteriology, Tarbiat Modares University, P.O. Box: 14115111, Tehran, Iran. Phone: +989120310214 E-mail: amin.talebi@modares.ac.ir Abstract Helicobacter pylori (H. pylori) is the pivotal cause of chronic gastritis, peptic ulcer diseases (PUD) and gastric cancer. Morphologically, the bacterium is spiral, Gram-negative and microaerophilic which survives lifespan in the human stomach in case of weak antibiotic therapy. There is a major difference in the pattern of global prevalence of H. pylori infection based on different levels of urbanization, hygiene, sanitation, access to clean water and other socioeconomic factors. To date, many studies have attempted to find significant associations between specific gastroduodenal diseases and dupA-positive strains, but no conclusive conclusion has been declared. The main reason for these inconsistent findings is the various methodologies applied in experiments which in turn have resulted in inaccurate observation. Our analysis showed that the existence of various alleles located in the dupA cluster would be a novel explanation for different associations found between this bacterial gene and diseases. In detailed experiments examining our proposed alleles using a large number of patients can be useful to disclose a significant clinical association between H. pylori dupA-positive strains and duodenal ulcer.
通讯作者:Amin Talebi Bezmin Abadi Tarbiat Modares大学细菌学系，邮政信箱:14115111，德黑兰，伊朗。摘要幽门螺杆菌(Helicobacter pylori, H. pylori)是慢性胃炎、消化性溃疡疾病(PUD)和胃癌的关键病因。在形态上，该细菌呈螺旋状，革兰氏阴性，嗜微气，在抗生素治疗薄弱的情况下在人胃中存活终生。根据城市化水平、个人卫生、环境卫生、获得清洁水和其他社会经济因素的不同，幽门螺杆菌感染的全球流行模式存在重大差异。迄今为止，许多研究试图发现特定胃十二指肠疾病与dupa阳性菌株之间的显著关联，但尚未宣布结论性结论。这些不一致的发现的主要原因是实验中应用的各种方法，而这些方法反过来又导致了不准确的观察。我们的分析表明，位于dupA簇的各种等位基因的存在将是这种细菌基因与疾病之间发现的不同关联的一种新的解释。在详细的实验中，使用大量患者检查我们提出的等位基因可能有助于揭示幽门螺杆菌dupa阳性菌株与十二指肠溃疡之间的显着临床关联。
{"title":"A report on Allelic Variation in Helicobacter pylori dupA: A viewpoint","authors":"Golzar Fatahi, A. Abadi","doi":"10.29252/RMM.5.3.1","DOIUrl":"https://doi.org/10.29252/RMM.5.3.1","url":null,"abstract":"Corresponding Author: Amin Talebi Bezmin Abadi Department of Bacteriology, Tarbiat Modares University, P.O. Box: 14115111, Tehran, Iran. Phone: +989120310214 E-mail: amin.talebi@modares.ac.ir Abstract Helicobacter pylori (H. pylori) is the pivotal cause of chronic gastritis, peptic ulcer diseases (PUD) and gastric cancer. Morphologically, the bacterium is spiral, Gram-negative and microaerophilic which survives lifespan in the human stomach in case of weak antibiotic therapy. There is a major difference in the pattern of global prevalence of H. pylori infection based on different levels of urbanization, hygiene, sanitation, access to clean water and other socioeconomic factors. To date, many studies have attempted to find significant associations between specific gastroduodenal diseases and dupA-positive strains, but no conclusive conclusion has been declared. The main reason for these inconsistent findings is the various methodologies applied in experiments which in turn have resulted in inaccurate observation. Our analysis showed that the existence of various alleles located in the dupA cluster would be a novel explanation for different associations found between this bacterial gene and diseases. In detailed experiments examining our proposed alleles using a large number of patients can be useful to disclose a significant clinical association between H. pylori dupA-positive strains and duodenal ulcer.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"5 1","pages":"1-4"},"PeriodicalIF":0.0,"publicationDate":"2017-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43071679","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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