F. Ketabchi, Faezeh Namazi, Nasrin Hadi, Farinaz Khosravian, Mirza mohammad Raisinia, Mansoor Salehi
1. Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran. 2. Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran. 3. Medical Genetics Research Center of Genome, Isfahan University of Medical Sciences, Isfahan, Iran. 4. Cellular, Molecular and Genetics Research Center, Isfahan University of Medical Sciences, Isfahan, Iran. 5. Department of Microbiology, Kerman Branch, Islamic Azad University, Kerman, Iran.
{"title":"The Effect of Dracocephalum kotschyi Alcoholic Extract on the BCL2 and BAX Expression in SKBR3 Cell Line","authors":"F. Ketabchi, Faezeh Namazi, Nasrin Hadi, Farinaz Khosravian, Mirza mohammad Raisinia, Mansoor Salehi","doi":"10.32598/RMM.7.3.15","DOIUrl":"https://doi.org/10.32598/RMM.7.3.15","url":null,"abstract":"1. Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran. 2. Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran. 3. Medical Genetics Research Center of Genome, Isfahan University of Medical Sciences, Isfahan, Iran. 4. Cellular, Molecular and Genetics Research Center, Isfahan University of Medical Sciences, Isfahan, Iran. 5. Department of Microbiology, Kerman Branch, Islamic Azad University, Kerman, Iran.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45422586","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}
Seyed Morteza Robatjazi, Sanaz Mahboudi, M. Zeinoddini
Materials and Methods: First, the effect of chemical composition in culture medium (the complex and defined) was investigated in DT389-IL2 expression in E.coli BL21 (DE3) containing pET-IDZ plasmid in shack flask and lab-bioreactor culture. To enrich the carbon source, we added glucose 6-8 g/L to the complex culture medium. The composition of the defined medium was enriched by adding amino acids (valine, 0.0502; phenylalanine, 0.0132; lysine, 0.0184; aspartic acid, 0.016; and serine, 0.0251 g/L). The growth cell was measured by the optical density method and the expression yield was investigated by SDS-PAGE.
{"title":"Production of Recombinant Denileukin Diftitox: Assessment in the Lab-scale Bioreactor","authors":"Seyed Morteza Robatjazi, Sanaz Mahboudi, M. Zeinoddini","doi":"10.32598/RMM.7.3.21","DOIUrl":"https://doi.org/10.32598/RMM.7.3.21","url":null,"abstract":"Materials and Methods: First, the effect of chemical composition in culture medium (the complex and defined) was investigated in DT389-IL2 expression in E.coli BL21 (DE3) containing pET-IDZ plasmid in shack flask and lab-bioreactor culture. To enrich the carbon source, we added glucose 6-8 g/L to the complex culture medium. The composition of the defined medium was enriched by adding amino acids (valine, 0.0502; phenylalanine, 0.0132; lysine, 0.0184; aspartic acid, 0.016; and serine, 0.0251 g/L). The growth cell was measured by the optical density method and the expression yield was investigated by SDS-PAGE.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47672366","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}
zeinab Daneshyar, H. Goli, B. Mirzaei, M. Rabie, M. Haghshenas
{"title":"Prevalence and clinical symptoms of Human Parainfluenza and Influenza infections in patients admitted to Mazandaran province health centers, 2019","authors":"zeinab Daneshyar, H. Goli, B. Mirzaei, M. Rabie, M. Haghshenas","doi":"10.32598/RMM.7.3.29","DOIUrl":"https://doi.org/10.32598/RMM.7.3.29","url":null,"abstract":"","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47129906","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}
Hydrogels have been used for biomedical applications in recent decades. They are a perfect candidate for regenerative medicine as they resemble the extracellular matrix of native tissues. In addition, their highly hydrated structure makes them a suitable choice for drug and other therapeutics delivery. Injectable hydrogels have increasingly gained attention due to their capability for homogeneous mixing with cells and therapeutic agents, minimally invasive administration, and perfect defect filling. In this review, we discuss various mechanisms which facilitate injectability of hydrogels, including in situ gelling liquids, injectable gels, and injectable particles. Then, we explore the biomedical applications of injectable hydrogels, including tissue engineering, therapeutic agent delivery, and medical devices.
{"title":"Injectable Hydrogels: A Review of Injectability Mechanisms and Biomedical Applications","authors":"A. Mellati, J. Akhtari","doi":"10.18502/RMM.V6I4.4799","DOIUrl":"https://doi.org/10.18502/RMM.V6I4.4799","url":null,"abstract":"Hydrogels have been used for biomedical applications in recent decades. They are a perfect candidate for regenerative medicine as they resemble the extracellular matrix of native tissues. In addition, their highly hydrated structure makes them a suitable choice for drug and other therapeutics delivery. Injectable hydrogels have increasingly gained attention due to their capability for homogeneous mixing with cells and therapeutic agents, minimally invasive administration, and perfect defect filling. In this review, we discuss various mechanisms which facilitate injectability of hydrogels, including in situ gelling liquids, injectable gels, and injectable particles. Then, we explore the biomedical applications of injectable hydrogels, including tissue engineering, therapeutic agent delivery, and medical devices.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44849988","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}
Background: The most malignant form of infiltrating astrocytoma, glioblastoma multiforme (GBM), is one of the most aggressive human cancers. Foretinib diminished GBM cell invasion by downregulating the expression of matrix metalloproteinase 2 (MMP2). Aim: The study aimed to examine the anti-tumor activity of foretinib and to test its effect on MMP2 expression in T98 cells. Materials and Methods: T98 cells were used as an experimental model of glioblastoma. The effect of foretinib on the expression of MMP2 was evaluated using quantitative real-time polymerase chain reaction. Thereafter, the effect of foretinib on the enzyme levels of MMP was examined by zymography. Statistical analyses were performed with GraphPad Prism software. Results: A reduction in the expression of MMP2 was observed with an increase in the concentration of foretinib. Conclusion: Foretinib treatment leads to downregulation of MMP-2 and has a positive effect on T98 cells. We believe that foretinib can be subjected to further clinical investigation to develop a treatment for GBM.
{"title":"Effect of Foretinib on Matrix Metalloproteinase-2 (MMP2) Expression in Glioblastoma","authors":"M. Fotoohi, Nasrin Hadi, Faezeh Namazi","doi":"10.18502/RMM.V6I4.4800","DOIUrl":"https://doi.org/10.18502/RMM.V6I4.4800","url":null,"abstract":"Background: The most malignant form of infiltrating astrocytoma, glioblastoma multiforme (GBM), is one of the most aggressive human cancers. Foretinib diminished GBM cell invasion by downregulating the expression of matrix metalloproteinase 2 (MMP2). \u0000Aim: The study aimed to examine the anti-tumor activity of foretinib and to test its effect on MMP2 expression in T98 cells. \u0000Materials and Methods: T98 cells were used as an experimental model of glioblastoma. The effect of foretinib on the expression of MMP2 was evaluated using quantitative real-time polymerase chain reaction. Thereafter, the effect of foretinib on the enzyme levels of MMP was examined by zymography. Statistical analyses were performed with GraphPad Prism software. \u0000Results: A reduction in the expression of MMP2 was observed with an increase in the concentration of foretinib. \u0000Conclusion: Foretinib treatment leads to downregulation of MMP-2 and has a positive effect on T98 cells. We believe that foretinib can be subjected to further clinical investigation to develop a treatment for GBM.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48689895","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}
Nafiseh Pourmahdi, M. Zeinoddini, Mohamad Javad Dehghan Esmatabadi, F. Sheikhi
Background: Yersinia pestis and Francisella tularensis cause plague and tularemia, which are known as diseases of the newborn and elderly, respectively. Immunological and culture-based detection methods of these bacteria are time-consuming, costly, complicated and require advanced equipment. We aimed to design and synthesize a gene structure as positive control for molecular detection of these bacteria. Materials and Method: Conserved regions of each bacterium were determined. A fragment containing the fopA and caf1 genes (conserved genes of F. tularensis and Y. pestis, respectively) was artificially synthesized, cloned into the pUC57 vector (pUCfopA- caf1), transformed into E. coli DH5α, and used in a multiplex PCR assay. The sensitivity of this assay was examined by serial dilution of the extracted plasmid, whereas the specificity was examined using genomes of Escherichia coli, Salmonella typhi, Enterobacter aerogenes, Vibrio cholerae as templates. Finally, PCR products were analyzed in agarose gel electrophoresis. Results: As expected, our analysis showed a clear dual band in the size range of 107 bp to 176 bp, confirming the presence of fopA and caf1 genes. Another 351 bp band was detected due to amplification being dependent on the forward primer of fopA and the reverse primer of caf1. Optimization of the PCR protocol reduced the amplification of this 351 bp band. The sensitivity of this assay was determined to be 36×10 −3ng/μl and the selectivity test confirmed the specificity of this method is appropriate for the detection of target genes. Conclusion: This multiplex PCR method could be used in research laboratories for identification of these important pathogens.
{"title":"Simple and Rapid Detection of Yersinia pestis and Francisella tularensis Using Multiplex-PCR: Molecular Detection of Yersinia pestis and Francisella tularensis","authors":"Nafiseh Pourmahdi, M. Zeinoddini, Mohamad Javad Dehghan Esmatabadi, F. Sheikhi","doi":"10.18502/RMM.V6I4.4801","DOIUrl":"https://doi.org/10.18502/RMM.V6I4.4801","url":null,"abstract":"Background: Yersinia pestis and Francisella tularensis cause plague and tularemia, which are known as diseases of the newborn and elderly, respectively. Immunological and culture-based detection methods of these bacteria are time-consuming, costly, complicated and require advanced equipment. We aimed to design and synthesize a gene structure as positive control for molecular detection of these bacteria. \u0000Materials and Method: Conserved regions of each bacterium were determined. A fragment containing the fopA and caf1 genes (conserved genes of F. tularensis and Y. pestis, respectively) was artificially synthesized, cloned into the pUC57 vector (pUCfopA- caf1), transformed into E. coli DH5α, and used in a multiplex PCR assay. The sensitivity of this assay was examined by serial dilution of the extracted plasmid, whereas the specificity was examined using genomes of Escherichia coli, Salmonella typhi, Enterobacter aerogenes, Vibrio cholerae as templates. Finally, PCR products were analyzed in agarose gel electrophoresis. \u0000Results: As expected, our analysis showed a clear dual band in the size range of 107 bp to 176 bp, confirming the presence of fopA and caf1 genes. Another 351 bp band was detected due to amplification being dependent on the forward primer of fopA and the reverse primer of caf1. Optimization of the PCR protocol reduced the amplification of this 351 bp band. The sensitivity of this assay was determined to be 36×10 −3ng/μl and the selectivity test confirmed the specificity of this method is appropriate for the detection of target genes. \u0000Conclusion: This multiplex PCR method could be used in research laboratories for identification of these important pathogens.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49581033","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}
Background: Breast cancer is a hormone-dependent malignancy that is associated with estrogen and progesterone interactions. The liver is the most important organ to be affected by the metastasis of breast cancer, which causes functional impairment. Aim: We compared levels of obesity, 17β-estradiol, and secreted proteins in postmenopausal women with breast cancer but without hepatic symptoms to those in healthy postmenopausal women. Materials and Methods: We recruited 105 postmenopausal women with breast cancer but without any clinical hepatic symptoms based on a physician’s diagnosis, and 105 healthy postmenopausal women. After taking blood samples, we separated the serum and determined the levels of alanine aminotransferase (ALT), enzyme aspartate aminotransferase (AST), sex hormone-binding globulin (SHBG), and 17β-estradiol using an enzyme-linked immunosorbent assay (ELISA). The results were statistically analyzed using SPSS. Results: The mean ages of the subjects in the cancer and control groups were 60.88 ± 0.85 and 55.56 ± 0.81 years, respectively. The exception ages (p=0.002), body mass index (BMI) values (p=0.033), serum glutamic oxaloacetic transaminase (SGOT) levels/AST levels (p=3.1*10−4), serum glutamic pyruvic transaminase (SGPT) levels/ALT levels(p=0.001), SHBG levels(p=0.014), and 17β-estradiol levels(p=0.003) in the serum differed significantly between the groups. Moreover, the mean serum 17β-estradiol (E2) levels and weights were higher in the cancer group than in the control group. Nevertheless, the mean serum levels of synthetic liver enzymes (SHBG, ALT, and AST) were lower in the cancer group than in the control group. Conclusion: In general, the postmenopausal cancer patients had higher serum estrogen levels and BMIs than their healthy counterparts. Furthermore, the levels of liver enzymes apparently decreased in the cancer group, probably owing to liver malfunction.
{"title":"Serum Liver Proteins and 17","authors":"Z. T. Fard, F. Rouhollah, N. Nafisi","doi":"10.18502/RMM.V6I4.4803","DOIUrl":"https://doi.org/10.18502/RMM.V6I4.4803","url":null,"abstract":"Background: Breast cancer is a hormone-dependent malignancy that is associated with estrogen and progesterone interactions. The liver is the most important organ to be affected by the metastasis of breast cancer, which causes functional impairment. \u0000Aim: We compared levels of obesity, 17β-estradiol, and secreted proteins in postmenopausal women with breast cancer but without hepatic symptoms to those in healthy postmenopausal women. \u0000Materials and Methods: We recruited 105 postmenopausal women with breast cancer but without any clinical hepatic symptoms based on a physician’s diagnosis, and 105 healthy postmenopausal women. After taking blood samples, we separated the serum and determined the levels of alanine aminotransferase (ALT), enzyme aspartate aminotransferase (AST), sex hormone-binding globulin (SHBG), and 17β-estradiol using an enzyme-linked immunosorbent assay (ELISA). The results were statistically analyzed using SPSS. \u0000Results: The mean ages of the subjects in the cancer and control groups were 60.88 ± 0.85 and 55.56 ± 0.81 years, respectively. The exception ages (p=0.002), body mass index (BMI) values (p=0.033), serum glutamic oxaloacetic transaminase (SGOT) levels/AST levels (p=3.1*10−4), serum glutamic pyruvic transaminase (SGPT) levels/ALT levels(p=0.001), SHBG levels(p=0.014), and 17β-estradiol levels(p=0.003) in the serum differed significantly between the groups. Moreover, the mean serum 17β-estradiol (E2) levels and weights were higher in the cancer group than in the control group. Nevertheless, the mean serum levels of synthetic liver enzymes (SHBG, ALT, and AST) were lower in the cancer group than in the control group. \u0000Conclusion: In general, the postmenopausal cancer patients had higher serum estrogen levels and BMIs than their healthy counterparts. Furthermore, the levels of liver enzymes apparently decreased in the cancer group, probably owing to liver malfunction.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48009464","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}
H. Mousavi, H. R. Nejad, M. Zandi, Amir Khodavirdipour
Background: Pseudomonas aeruginosa is the primary cause of infection with impaired defense mechanisms. P. aeruginosa commonly causes nosocomial infections and is the most common pathogen isolated from patients hospitalized for longer than 1 week. We examined the antimicrobial effect of multilayered carbon nanotubes on multi-drug-resistant. Materials and Methods: In this research, 20 clinical isolates collected at Motahari Hospital (Tehran, Iran) were compared with the standard (ATCC 27853) and identified as P. aeruginosa based on biochemical testing. Conventional disk diffusion assay demonstrated the methicillin resistance of the isolates. Minimal inhibitory concentrations for antibiotics and the multilayer CNTs were determined using the microdilution method. Single-walled CNTs were prepared and their efficacy and potential synergism with antibiotics was assessed. Results: Synergism against P. aeruginosa was evident for methicillin + single-walled CNTs. Conclusion: The inhibitory effect of single-walled CNTs and methicillin was synergistic against the growth of P. aeruginosa.
{"title":"Antimicrobial Effect of Multilayered Carbon Nanotubes on Multi-Drug-Resistant Pseudomonas aeruginosa","authors":"H. Mousavi, H. R. Nejad, M. Zandi, Amir Khodavirdipour","doi":"10.18502/RMM.V6I4.4802","DOIUrl":"https://doi.org/10.18502/RMM.V6I4.4802","url":null,"abstract":"Background: Pseudomonas aeruginosa is the primary cause of infection with impaired defense mechanisms. P. aeruginosa commonly causes nosocomial infections and is the most common pathogen isolated from patients hospitalized for longer than 1 week. We examined the antimicrobial effect of multilayered carbon nanotubes on multi-drug-resistant. \u0000Materials and Methods: In this research, 20 clinical isolates collected at Motahari Hospital (Tehran, Iran) were compared with the standard (ATCC 27853) and identified as P. aeruginosa based on biochemical testing. Conventional disk diffusion assay demonstrated the methicillin resistance of the isolates. Minimal inhibitory concentrations for antibiotics and the multilayer CNTs were determined using the microdilution method. Single-walled CNTs were prepared and their efficacy and potential synergism with antibiotics was assessed. \u0000Results: Synergism against P. aeruginosa was evident for methicillin + single-walled CNTs. \u0000Conclusion: The inhibitory effect of single-walled CNTs and methicillin was synergistic against the growth of P. aeruginosa.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49251359","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}
N. Boroumand, Amir Eshaghiyan, P. Behshood, Behnaz Nateghi, F. Emadi
Background: Multiple sclerosis (MS) is an autoimmune disease that causes chronic inflammation of the central nervous system. MicroRNAs (miRNAs) are small non-coding RNAs 19–24 nucleotides long, which are differentially expressed in different tissues. The role of miRNAs in MS remains unclear. Aim: We assessed miR-10a transcript levels in MS patients during recurrence and two months after relapse. Materials and Methods: In this case-control study, we used real-time PCR to examine miR-10a expression in the peripheral blood mononuclear cells of 60 patients with relapsing-remitting multiple sclerosis (RRMS), 30 during recurrence and 30 two months after relapse, and 30 healthy subjects who were referred to the MS Clinic of Kashani Hospital, Isfahan Province. In silico analysis was also performed on the validated miR- 10a targets using miRTarBase. Results: miR-10a expression was higher in RRMS patients during recurrence and two months after relapse (p < 0.0001 and p < 0.0001, respectively) than in the healthy subjects. Furthermore, in silico molecular signaling enrichment analysis identified 12 mRNAs as validated miR-10a targets. Conclusion: The expression of miR-10a was elevated in patients with RRMS compared to healthy subjects, suggesting that miR-10a could be a potential biomarker for RRMS diagnosis.
{"title":"Increased Circulating miR-10a Levels Associated with Multiple Sclerosis","authors":"N. Boroumand, Amir Eshaghiyan, P. Behshood, Behnaz Nateghi, F. Emadi","doi":"10.18502/RMM.V6I4.4804","DOIUrl":"https://doi.org/10.18502/RMM.V6I4.4804","url":null,"abstract":"Background: Multiple sclerosis (MS) is an autoimmune disease that causes chronic inflammation of the central nervous system. MicroRNAs (miRNAs) are small non-coding RNAs 19–24 nucleotides long, which are differentially expressed in different tissues. The role of miRNAs in MS remains unclear. \u0000Aim: We assessed miR-10a transcript levels in MS patients during recurrence and two months after relapse. \u0000Materials and Methods: In this case-control study, we used real-time PCR to examine miR-10a expression in the peripheral blood mononuclear cells of 60 patients with relapsing-remitting multiple sclerosis (RRMS), 30 during recurrence and 30 two months after relapse, and 30 healthy subjects who were referred to the MS Clinic of Kashani Hospital, Isfahan Province. In silico analysis was also performed on the validated miR- 10a targets using miRTarBase. \u0000Results: miR-10a expression was higher in RRMS patients during recurrence and two months after relapse (p < 0.0001 and p < 0.0001, respectively) than in the healthy subjects. Furthermore, in silico molecular signaling enrichment analysis identified 12 mRNAs as validated miR-10a targets. \u0000Conclusion: The expression of miR-10a was elevated in patients with RRMS compared to healthy subjects, suggesting that miR-10a could be a potential biomarker for RRMS diagnosis.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48040721","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}
Background: L-Asparaginase (L-Asp) is used as an efficient anti-cancer drug, especially for acute lymphoblastic leukemia (ALL). Currently, two bacterial asparaginase isoenzymes are used for cancer treatment. Therefore, this research focused on isolating native bacteria with the ability to produce L-Asp. Materials and Methods: L-Asp producing bacteria were isolated from soil samples on 9K medium supplemented with L-Asp as nitrogen source. Detection of L-Asp activity was performed by observing color change of the agar medium from yellow to orange due to the release of ammonia around the colonies. After the isolation and identification of the bacterium, L-Asp production was first optimized by the one factor-at-the-time (OFAT) technique followed by the response surface method. Next, the enzyme was extracted, purified, and assessed for antileukemia activity on U937 and MRC-5 cell lines. Results: The results revealed that L-Asp produced by Rouxiella sp. AF1 significantly inhibited the growth of U937 cells at a dose of up to 0.04 IU/ml, while MRC-5 was not affected at any enzyme doses. The final purification of the enzyme was achieved by column chromatography (Sephadex G-100) at approximately 0.31 mg/ml, and its specific activity was determined to be 0.51 IU/mg. The OFAT optimization experiments were performed primarily to determine optimal enzyme conditions, which were found to be neutral pH (pH7), 30 ∘C temperature, and 3 % NaCl, 1 % peptone, and 1% glucose concentrations. Statistical optimization was based on five factors obtained from OFAT, and response surface method (RSM) analysis introduced a quadratic model for enzyme production at the optimal range of these variables. This model provided an equation for measuring the effect of physiochemical conditions on final enzyme production. Conclusion: We showed that native bacteria may be novel candidates for isolating new metabolites such as L-Asp. Because many bacteria grow in unknown environments with unique ecological properties, the probability of discovering novel bacterial species producing bioactive compounds is high.
{"title":"L-Asparaginase-producing Rouxiella Species Isolation, Antileukemia Activity Evaluation, and Enzyme Production Optimization","authors":"F. Gilavand, A. Kavyanifard, A. Marzban","doi":"10.18502/RMM.V6I3.4608","DOIUrl":"https://doi.org/10.18502/RMM.V6I3.4608","url":null,"abstract":"Background: L-Asparaginase (L-Asp) is used as an efficient anti-cancer drug, especially for acute lymphoblastic leukemia (ALL). Currently, two bacterial asparaginase isoenzymes are used for cancer treatment. Therefore, this research focused on isolating native bacteria with the ability to produce L-Asp. Materials and Methods: L-Asp producing bacteria were isolated from soil samples on 9K medium supplemented with L-Asp as nitrogen source. Detection of L-Asp activity was performed by observing color change of the agar medium from yellow to orange due to the release of ammonia around the colonies. After the isolation and identification of the bacterium, L-Asp production was first optimized by the one factor-at-the-time (OFAT) technique followed by the response surface method. Next, the enzyme was extracted, purified, and assessed for antileukemia activity on U937 and MRC-5 cell lines. Results: The results revealed that L-Asp produced by Rouxiella sp. AF1 significantly inhibited the growth of U937 cells at a dose of up to 0.04 IU/ml, while MRC-5 was not affected at any enzyme doses. The final purification of the enzyme was achieved by column chromatography (Sephadex G-100) at approximately 0.31 mg/ml, and its specific activity was determined to be 0.51 IU/mg. The OFAT optimization experiments were performed primarily to determine optimal enzyme conditions, which were found to be neutral pH (pH7), 30 ∘C temperature, and 3 % NaCl, 1 % peptone, and 1% glucose concentrations. Statistical optimization was based on five factors obtained from OFAT, and response surface method (RSM) analysis introduced a quadratic model for enzyme production at the optimal range of these variables. This model provided an equation for measuring the effect of physiochemical conditions on final enzyme production. Conclusion: We showed that native bacteria may be novel candidates for isolating new metabolites such as L-Asp. Because many bacteria grow in unknown environments with unique ecological properties, the probability of discovering novel bacterial species producing bioactive compounds is high.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2019-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45124750","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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