Introduction: Proliferation of spermatogonial stem cells (SSCs) can be a treatment for infertile men. Here, we design an efficient method based on culturing in the presence of Sertoli cells to improve the expression level of some specific spermatogonia stem cell genes during two weeks post culture. Materials and Methods: Cells were derived from neonatal (2-6 days old) mice testes and were cultured in DMEM medium with FBS. The colonization of cultured SSCs in days 4, 7, and 14 of culture was counted via phase-contrast microscope and Image J software. Methyl thiazolyl tetrazolium (MTT) test was performed to evaluate the viability of cultured SSCs in days 3, 7, and 14 of culture. The expression level and the alteration pattern of specific spermatogonial markers, i.e., Stra8, DAZL, and Piwill2 was examined via real-time polymerase chain reaction (PCR) during two weeks post culture. Results: The number and the diameters of colonies showed a significant increase in cultured cells. MTT results proved the higher viability of testicular cells during the culture period. The results of ALP staining detected a positive reaction in spermatogonia colonies. Real-time PCR data showed that culturing SSCs in the presence of interstitial cells of the testis, amplified the level and alteration pattern of specific spermatogonia stem cells genes beneficial in the enrichment of SSCs propagation. Conclusion: Providing a similar culture environment to testicular niche increases viability, forms SSCs colonies, and regulates the level and alteration pattern of spermatogonia stem cell genes.
{"title":"An Efficient In Vitro Culture System To Amplify Spermatogonia Stem Cell Markers","authors":"Z. Narimanpour, M. Nazm Bojnordi, H. Ghasemi","doi":"10.32598/RMM.8.3.2","DOIUrl":"https://doi.org/10.32598/RMM.8.3.2","url":null,"abstract":"Introduction: Proliferation of spermatogonial stem cells (SSCs) can be a treatment for infertile men. Here, we design an efficient method based on culturing in the presence of Sertoli cells to improve the expression level of some specific spermatogonia stem cell genes during two weeks post culture. Materials and Methods: Cells were derived from neonatal (2-6 days old) mice testes and were cultured in DMEM medium with FBS. The colonization of cultured SSCs in days 4, 7, and 14 of culture was counted via phase-contrast microscope and Image J software. Methyl thiazolyl tetrazolium (MTT) test was performed to evaluate the viability of cultured SSCs in days 3, 7, and 14 of culture. The expression level and the alteration pattern of specific spermatogonial markers, i.e., Stra8, DAZL, and Piwill2 was examined via real-time polymerase chain reaction (PCR) during two weeks post culture. Results: The number and the diameters of colonies showed a significant increase in cultured cells. MTT results proved the higher viability of testicular cells during the culture period. The results of ALP staining detected a positive reaction in spermatogonia colonies. Real-time PCR data showed that culturing SSCs in the presence of interstitial cells of the testis, amplified the level and alteration pattern of specific spermatogonia stem cells genes beneficial in the enrichment of SSCs propagation. Conclusion: Providing a similar culture environment to testicular niche increases viability, forms SSCs colonies, and regulates the level and alteration pattern of spermatogonia stem cell genes.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"117-124"},"PeriodicalIF":0.0,"publicationDate":"2020-08-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49418408","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: Lactoferrin is a glycoprotein with antimicrobial, antioxidant, immune-modulating, antiviral, and most importantly anticancer properties. In the present study, the effect of lactoferrin on breast cancer cell growth and the expression of Bax and Bak genes are evaluated. Materials and Methods: MCF7 cells were cultured in a 96-well plate with 1×105 cells in each well. Different lactoferrin concentrations of 0, 50, 300, 600, and 800 μg/mL were added to each well in three replicates and the well was incubated for 24 hours. After treatment, cell survival was measured using the MTT assay. To determine the level of expression of Bax and Bak genes, the cells were treated with lactoferrin concentrations of 0, 50, and 800 μg/mL in 2 replicates for 24 hours. Then RNA extraction was performed and cDNA was synthesized immediately and the expression of the genes in the presence of beta-actin reference gene and cyber-green fluorescence color was investigated with real-time reactions. Results: The cells viability in lactoferrin concentrations of 0, 50, 300, 500 and 800 μg/μL were 100%, 94%, 83%, 62%, and 32%, respectively. The expression level of the Bax gene at a concentration of 50 μg increased by 2.71 times and in 800 μg concentration decreased by 0.88 times. Also, the expression level of the Bak gene at concentrations of 50 and 800 μg increased by 1.23 and 1.0 fold, respectively. Statistical analysis of the data indicated that the expression levels of two genes at a concentration of 50 μg/mL of lactoferrin significantly increased (P<0.01), compared to the control. The significance level in this study was set at < 0.05. Conclusion: In this study, lactoferrin showed a growth inhibitory effect on breast cancer cells and increased the expression of Bax and Bak genes involved in apoptosis at a concentration of 50 µg/mL.
{"title":"Anticancer Effect of Bovine Lactoferrin on Breast Cancer Cell Line MCF7 and the Evaluation of Bax and Bak Genes Expression Involved in Apoptosis","authors":"Amir Khalafi, F. Moradian, A. Rafiei","doi":"10.32598/RMM.8.3.726.3","DOIUrl":"https://doi.org/10.32598/RMM.8.3.726.3","url":null,"abstract":"Background: Lactoferrin is a glycoprotein with antimicrobial, antioxidant, immune-modulating, antiviral, and most importantly anticancer properties. In the present study, the effect of lactoferrin on breast cancer cell growth and the expression of Bax and Bak genes are evaluated. Materials and Methods: MCF7 cells were cultured in a 96-well plate with 1×105 cells in each well. Different lactoferrin concentrations of 0, 50, 300, 600, and 800 μg/mL were added to each well in three replicates and the well was incubated for 24 hours. After treatment, cell survival was measured using the MTT assay. To determine the level of expression of Bax and Bak genes, the cells were treated with lactoferrin concentrations of 0, 50, and 800 μg/mL in 2 replicates for 24 hours. Then RNA extraction was performed and cDNA was synthesized immediately and the expression of the genes in the presence of beta-actin reference gene and cyber-green fluorescence color was investigated with real-time reactions. Results: The cells viability in lactoferrin concentrations of 0, 50, 300, 500 and 800 μg/μL were 100%, 94%, 83%, 62%, and 32%, respectively. The expression level of the Bax gene at a concentration of 50 μg increased by 2.71 times and in 800 μg concentration decreased by 0.88 times. Also, the expression level of the Bak gene at concentrations of 50 and 800 μg increased by 1.23 and 1.0 fold, respectively. Statistical analysis of the data indicated that the expression levels of two genes at a concentration of 50 μg/mL of lactoferrin significantly increased (P<0.01), compared to the control. The significance level in this study was set at < 0.05. Conclusion: In this study, lactoferrin showed a growth inhibitory effect on breast cancer cells and increased the expression of Bax and Bak genes involved in apoptosis at a concentration of 50 µg/mL.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"107-116"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46376828","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. Habibi, Noorsaadat Saajedi, N. Ghahtan, Saajedeh Habibi
Background: Due to increased bacterial resistance to common antibiotics, the tendency towards using herbal extracts is increasing. Plantago major L, Plantago ovata, Lallemantia iberica L, Sinapis arvensis L, and Ficus carica are widely used as herbal plants in traditional medicine. They were known to have a variety of therapeutic effects. The current study aimed to evaluate the antibacterial activity of hydroalcoholic extract of these herbs against some hospital-acquired infections. Materials and Methods: Disk-diffusion antibiotic sensitivity testing, minimum inhibitory concentration, and minimum bactericidal concentration of hydroalcoholic extracts were applied to assess the antibacterial activity compared with tetracycline, as a control antibiotic. Results: The results of this experiment showed that the L. iberica and S. arvensis extract had the greatest effect on Pseudomonas aeruginosa, Staphylococcus aureus, and Proteus vulgaris. All the tested medicinal plants had a high antibacterial effect on P. vulgaris, except P. ovata. Conclusion: The results of this study show that the replacement of chemical drugs with herbal extract could be effective in the elimination of bacterial growth.
背景：由于细菌对常见抗生素的耐药性增加，使用草药提取物的趋势也在增加。Plantago major L、Plantago ovata、Lallemania iberica L、Sinapis arvensis L和Ficus carica是传统医学中广泛使用的草药植物。众所周知，它们具有多种治疗作用。本研究旨在评估这些草药的水醇提取物对一些医院获得性感染的抗菌活性。材料与方法：采用纸片扩散法测定水醇提取物的抗生素敏感性、最低抑菌浓度和最低杀菌浓度，并与四环素作为对照抗生素进行比较。结果：本实验结果表明，白藜芦醇和山葡萄提取物对铜绿假单胞菌、金黄色葡萄球菌和普通变形杆菌的影响最大。除卵形假单胞菌外，所有受试药用植物对寻常假单胞菌均具有较高的抗菌作用。结论：本研究结果表明，用草药提取物代替化学药物可以有效地消除细菌生长。
{"title":"Antibacterial Effect of Hydroalcoholic Extracts of Herbal Plants Against Some Hospital-Acquired Infections","authors":"H. Habibi, Noorsaadat Saajedi, N. Ghahtan, Saajedeh Habibi","doi":"10.32598/RMM.8.3.1","DOIUrl":"https://doi.org/10.32598/RMM.8.3.1","url":null,"abstract":"Background: Due to increased bacterial resistance to common antibiotics, the tendency towards using herbal extracts is increasing. Plantago major L, Plantago ovata, Lallemantia iberica L, Sinapis arvensis L, and Ficus carica are widely used as herbal plants in traditional medicine. They were known to have a variety of therapeutic effects. The current study aimed to evaluate the antibacterial activity of hydroalcoholic extract of these herbs against some hospital-acquired infections. Materials and Methods: Disk-diffusion antibiotic sensitivity testing, minimum inhibitory concentration, and minimum bactericidal concentration of hydroalcoholic extracts were applied to assess the antibacterial activity compared with tetracycline, as a control antibiotic. Results: The results of this experiment showed that the L. iberica and S. arvensis extract had the greatest effect on Pseudomonas aeruginosa, Staphylococcus aureus, and Proteus vulgaris. All the tested medicinal plants had a high antibacterial effect on P. vulgaris, except P. ovata. Conclusion: The results of this study show that the replacement of chemical drugs with herbal extract could be effective in the elimination of bacterial growth.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"133-138"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44218477","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}
Roya Abbasinatajomrani, D. Qujeq, V. Hosseini, R. Hajihosseini
Background: Currently, glycans, which are known as functional molecules in the biological system, are being under study as potential cancer markers. This study aimed to determine the level of serum L-fucose and sialic acid as the biomarkers in patients with colorectal cancer (CRC). Materials and Methods: The patients with CRC (n=40, 20 men and 20 women) participated in the present study. The spectrophotometric method was used to measure the levels of L-fucose and sialic acid in the serum of the patients. SPSS (version 21) was used to analyze the obtained data. The results were expressed as mean ± SD. Results: The mean ± SD L-fucose level in patients with CRC was 27.46 ± 4.8 ng/mL, which was more than this level in the healthy control group (18.64±3.1 ng/mL). Also, the mean ± SD serum concentration of sialic acid in patients with CRC was 2.1 ± 0.41 ng/mL, which was more than the mean ± SD sialic acid level of 1.23±0.21 ng/mL in the healthy controls. Conclusion: Serum concentration of L-fucose and sialic acid increased significantly (P < 0.05) in patients with CRC compared with the healthy controls. We believe that determining serum L-fucose and sialic acid levels could be useful for the detection of CRC patients in the early stage.
{"title":"Evaluation of L-fucose and Sialic Acid Levels in Patients With Colorectal Cancer and Control Subject","authors":"Roya Abbasinatajomrani, D. Qujeq, V. Hosseini, R. Hajihosseini","doi":"10.32598/RMM.8.3.609.7","DOIUrl":"https://doi.org/10.32598/RMM.8.3.609.7","url":null,"abstract":"Background: Currently, glycans, which are known as functional molecules in the biological system, are being under study as potential cancer markers. This study aimed to determine the level of serum L-fucose and sialic acid as the biomarkers in patients with colorectal cancer (CRC). Materials and Methods: The patients with CRC (n=40, 20 men and 20 women) participated in the present study. The spectrophotometric method was used to measure the levels of L-fucose and sialic acid in the serum of the patients. SPSS (version 21) was used to analyze the obtained data. The results were expressed as mean ± SD. Results: The mean ± SD L-fucose level in patients with CRC was 27.46 ± 4.8 ng/mL, which was more than this level in the healthy control group (18.64±3.1 ng/mL). Also, the mean ± SD serum concentration of sialic acid in patients with CRC was 2.1 ± 0.41 ng/mL, which was more than the mean ± SD sialic acid level of 1.23±0.21 ng/mL in the healthy controls. Conclusion: Serum concentration of L-fucose and sialic acid increased significantly (P < 0.05) in patients with CRC compared with the healthy controls. We believe that determining serum L-fucose and sialic acid levels could be useful for the detection of CRC patients in the early stage.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"147-152"},"PeriodicalIF":0.0,"publicationDate":"2020-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49600633","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}
Gholamreza Farnoosh, K. Hassanpour, T. Badri, Reza Hosseiniara
On March 11, 2020, the World Health Organization (WHO) announced the pandemic outbreak of coronavirus disease 2019 (COVID-19), due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which began in December 2019 in Wuhan, China (1). Iran was the 25th country affected by COVID-19. According to the Iranian Ministry of Health, every 12 minutes, one person dies in Iran because of COVID-19. After a 3.5-fold and 12.9-fold increase in the number of COVID-19 affected people and deaths, respectively in the second two weeks compared with the first two weeks after the official announcement of COVID-19 on February 19, 2020, in Iran (2), the coincidence of this epidemic with the celebration of New Year Eve, may lead to a human and health catastrophe in Iran. Nowruz (New Year Eve) is the first day of the Iranian New Year on March 20 (or the previous or following day). It marks the beginning of spring in the Northern Hemisphere and takes place with celebrations. Before the start of the New Year in Iran on March 20, all 31 provinces were affected by COVID-19, with some provinces including Qom, Tehran, Guilan reporting the highest prevalence rates (2). The preparations for the celebration of New Year's Eve in Iran include shopping in crowded markets and beginning of the New Year Eve holiday in Iran (6 official days for administrations and 13 days for schools and universities) is along with traveling, that accelerates human-to-human transmission, effectively spreading SARS-CoV-2 (3). Due to the outbreak of COVID-19 during Nowruz, abroad travel had decreased, but domestic travel in Iran had not stopped. Despite the closing of museums and recreation centers, and repeated official announcements that no amenities will be offered, Nowruz trips continued. Besides domestic tourism attractions in Iran as a four-season country, many people in big cities are returned to their villages and hometowns for the New Year's holiday to visit their parents or grandparents. These visits can lead to an increased prevalence of COVID-19 in adults, which is dangerous and worrying. This year, with the beginning of Nowruz, major highways and tourist areas were hit by heavy traffic, too. Nowruz trips of three million Iranians from the 13 affected provinces with COVID-19 (during March 17-20, 2020) demonstrated that many people refused to stay at home, while Iranian authorities and the WHO have repeatedly urged people not to travel, in order to prevent the spread of SARS-CoV-2 (2). Quarantine of cities with COVID-19 cases and controlling citizens' traffic to these cities are strategies recommended by WHO to control the outbreak of SARS-CoV-2, and China's experience as the main sources of this outbreak has confirmed the effectiveness of the measure (4). But no quarantine in any city had been implemented in Iran. Although some traffic restrictions had been put in place, they were not fully enforced in any province. According to the Iranian Ministry of Roads and Urban Development, t
{"title":"What Are the Consequences When the New Year Eve in Iran Coincides With COVID-19?","authors":"Gholamreza Farnoosh, K. Hassanpour, T. Badri, Reza Hosseiniara","doi":"10.32598/RMM.8.2.1145.1","DOIUrl":"https://doi.org/10.32598/RMM.8.2.1145.1","url":null,"abstract":"On March 11, 2020, the World Health Organization (WHO) announced the pandemic outbreak of coronavirus disease 2019 (COVID-19), due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which began in December 2019 in Wuhan, China (1). Iran was the 25th country affected by COVID-19. According to the Iranian Ministry of Health, every 12 minutes, one person dies in Iran because of COVID-19. After a 3.5-fold and 12.9-fold increase in the number of COVID-19 affected people and deaths, respectively in the second two weeks compared with the first two weeks after the official announcement of COVID-19 on February 19, 2020, in Iran (2), the coincidence of this epidemic with the celebration of New Year Eve, may lead to a human and health catastrophe in Iran. Nowruz (New Year Eve) is the first day of the Iranian New Year on March 20 (or the previous or following day). It marks the beginning of spring in the Northern Hemisphere and takes place with celebrations. Before the start of the New Year in Iran on March 20, all 31 provinces were affected by COVID-19, with some provinces including Qom, Tehran, Guilan reporting the highest prevalence rates (2). The preparations for the celebration of New Year's Eve in Iran include shopping in crowded markets and beginning of the New Year Eve holiday in Iran (6 official days for administrations and 13 days for schools and universities) is along with traveling, that accelerates human-to-human transmission, effectively spreading SARS-CoV-2 (3). Due to the outbreak of COVID-19 during Nowruz, abroad travel had decreased, but domestic travel in Iran had not stopped. Despite the closing of museums and recreation centers, and repeated official announcements that no amenities will be offered, Nowruz trips continued. Besides domestic tourism attractions in Iran as a four-season country, many people in big cities are returned to their villages and hometowns for the New Year's holiday to visit their parents or grandparents. These visits can lead to an increased prevalence of COVID-19 in adults, which is dangerous and worrying. This year, with the beginning of Nowruz, major highways and tourist areas were hit by heavy traffic, too. Nowruz trips of three million Iranians from the 13 affected provinces with COVID-19 (during March 17-20, 2020) demonstrated that many people refused to stay at home, while Iranian authorities and the WHO have repeatedly urged people not to travel, in order to prevent the spread of SARS-CoV-2 (2). Quarantine of cities with COVID-19 cases and controlling citizens' traffic to these cities are strategies recommended by WHO to control the outbreak of SARS-CoV-2, and China's experience as the main sources of this outbreak has confirmed the effectiveness of the measure (4). But no quarantine in any city had been implemented in Iran. Although some traffic restrictions had been put in place, they were not fully enforced in any province. According to the Iranian Ministry of Roads and Urban Development, t","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"49-50"},"PeriodicalIF":0.0,"publicationDate":"2020-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47778615","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}
Mohamad Najarasl, M. Zeinoddini, A. Saeeidinia, Reza Hasan Sajedi
BackgroundDenileukin diftitox (trade name, Ontak) is the first recombinant immunotoxin (IM), in which the binding domain of diphtheria toxin has been replaced by the amino acid sequence of human interleukin-2 (DT389IL-2) using genetic engineering. Purity, stability, and structural property of the protein are critical factors for the scale-up production of this fusion protein. In this IM, location 519 has free cysteine residue that leads to cross S-S bound formation in the refolding process and, as a result, misfolding/aggregation of the protein may occur.Materials and MethodsTo inhibit misfolding/aggregation, we substituted cysteine 519 by a serine residue with site-directed mutagenesis, and then the ability of the mutated protein for binding to the IL-2 receptor was predicted and determined by bioinformatics tools. For this purpose, the sequence of the denileukin diftitox was adopted from Drugbank, and the mentioned substitution applied. Two methods determined the folding of the fusion protein: de novo modeling method (by utilizing the I-TASSER database) and homology modeling method (by using some databases and tools, including Swiss-Model, PHYRE2, M4T, ModWeb, RaptorX, and EasyModeller). Finally, the ability of the proteins for binding to the IL-2 receptor was investigated by pyDock and Zdock docking servers, as well as Hex software.ResultsThe result showed that the mutated form (C519S) of this protein folds appropriately, and the ΔG of the models, measured by STRUM, showed no significant variation. Also, docking analysis has shown that the protein can efficiently bind to the IL-2 receptor without any substantial changes in the binding energy.ConclusionThe present study shows that the suggested mutation of this protein can be an acceptable replacement for denileukin diftitox with a similar affinity and a more proper refolding process.
{"title":"Molecular Docking and In Silico Study of Denileukin Diftitox: Comparison of Wild Type With C519S Mutant","authors":"Mohamad Najarasl, M. Zeinoddini, A. Saeeidinia, Reza Hasan Sajedi","doi":"10.32598/RMM.8.2.903.6","DOIUrl":"https://doi.org/10.32598/RMM.8.2.903.6","url":null,"abstract":"BackgroundDenileukin diftitox (trade name, Ontak) is the first recombinant immunotoxin (IM), in which the binding domain of diphtheria toxin has been replaced by the amino acid sequence of human interleukin-2 (DT389IL-2) using genetic engineering. Purity, stability, and structural property of the protein are critical factors for the scale-up production of this fusion protein. In this IM, location 519 has free cysteine residue that leads to cross S-S bound formation in the refolding process and, as a result, misfolding/aggregation of the protein may occur.Materials and MethodsTo inhibit misfolding/aggregation, we substituted cysteine 519 by a serine residue with site-directed mutagenesis, and then the ability of the mutated protein for binding to the IL-2 receptor was predicted and determined by bioinformatics tools. For this purpose, the sequence of the denileukin diftitox was adopted from Drugbank, and the mentioned substitution applied. Two methods determined the folding of the fusion protein: de novo modeling method (by utilizing the I-TASSER database) and homology modeling method (by using some databases and tools, including Swiss-Model, PHYRE2, M4T, ModWeb, RaptorX, and EasyModeller). Finally, the ability of the proteins for binding to the IL-2 receptor was investigated by pyDock and Zdock docking servers, as well as Hex software.ResultsThe result showed that the mutated form (C519S) of this protein folds appropriately, and the ΔG of the models, measured by STRUM, showed no significant variation. Also, docking analysis has shown that the protein can efficiently bind to the IL-2 receptor without any substantial changes in the binding energy.ConclusionThe present study shows that the suggested mutation of this protein can be an acceptable replacement for denileukin diftitox with a similar affinity and a more proper refolding process.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"83-90"},"PeriodicalIF":0.0,"publicationDate":"2020-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49249461","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}
Zahra Norouzi Bazgir, B. Mirzaei, M. Haghshenas, H. Goli, Ebrahim Shafaie
BackgroundMulti-drug resistant (MDR) Citrobacter freundii (C. freundii) as a causative agent of nosocomial infections is a health threat, especially in hospitals. This study was conducted to determine the prevalence of MDR C. freundii, considering isolation sites and a variety of utilized antibiotics.Materials and MethodsIn this cross-sectional study, the clinical samples of C. freundii strains were collected and screened using traditional bacteriological tests in Zareh Hospital, Sari City, Iran, during 2016-2017. We used disk diffusion methods to assess the susceptibility patterns of isolates according to the Clinical Laboratory Standard Institute (CLSI) guidelines.ResultsOut of 3248 clinical samples, C. freundii strains were detected in 109 samples (32.1% females and 67.9% males). Susceptibility tests indicated that 89 isolates (81.65%) were MDR strains. Frequencies of MDR C. freundii strains were higher in the Behavioral Intensive Care Unit (BICU) (37.61%) and restoration ward (29.35%) compared with other hospital wards.ConclusionConsidering the MDR C. freundii strains detected from burn hospital wards, it is necessary to implement prevention criteria for their eradication from burn hospitals. The results indicate the urgent need to design more practical methods for controlling infection in hospital wards.
{"title":"Multi-drug Resistant Citrobacter freundii Isolates in a Burn Hospital in Northeast of Iran: A Single-Center Cross-sectional Study","authors":"Zahra Norouzi Bazgir, B. Mirzaei, M. Haghshenas, H. Goli, Ebrahim Shafaie","doi":"10.32598/RMM.8.2.893.1","DOIUrl":"https://doi.org/10.32598/RMM.8.2.893.1","url":null,"abstract":"BackgroundMulti-drug resistant (MDR) Citrobacter freundii (C. freundii) as a causative agent of nosocomial infections is a health threat, especially in hospitals. This study was conducted to determine the prevalence of MDR C. freundii, considering isolation sites and a variety of utilized antibiotics.Materials and MethodsIn this cross-sectional study, the clinical samples of C. freundii strains were collected and screened using traditional bacteriological tests in Zareh Hospital, Sari City, Iran, during 2016-2017. We used disk diffusion methods to assess the susceptibility patterns of isolates according to the Clinical Laboratory Standard Institute (CLSI) guidelines.ResultsOut of 3248 clinical samples, C. freundii strains were detected in 109 samples (32.1% females and 67.9% males). Susceptibility tests indicated that 89 isolates (81.65%) were MDR strains. Frequencies of MDR C. freundii strains were higher in the Behavioral Intensive Care Unit (BICU) (37.61%) and restoration ward (29.35%) compared with other hospital wards.ConclusionConsidering the MDR C. freundii strains detected from burn hospital wards, it is necessary to implement prevention criteria for their eradication from burn hospitals. The results indicate the urgent need to design more practical methods for controlling infection in hospital wards.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"63-70"},"PeriodicalIF":0.0,"publicationDate":"2020-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"47216580","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}
Atefeh Verdi, S. Sahraei, E. Asa, Rahil Jannatifar, Mohamad Bagher Masaeimanesh
BackgroundOne of the main causes of male infertility is the negative effects of oxidative stress. Follicle-stimulating hormone (FSH) plays an essential role in spermatogenesis, as well as in the maintenance of sperm DNA integrity. This study aimed to determine whether the recombinant human follicle-stimulating hormone (rhFSH) treatment of sperm parameters could positively affect sperm DNA and oxidative DNA fragmentation in oligozoospermia infertile men.Materials and MethodsThis interventional study was carried out on a sample of 50 oligozoospermia infertile men. To this end, sperm DNA fragmentation and ROS as an oxidative stress marker were measured before and after treatment with the rhFSH sperm parameters.ResultsThe sperm parameters (concentration, mobility, and morphology) were significantly different in the oligozoospermia infertile patients before and after the rhFSH treatment (p<0.05). Moreover, sperm DNA fragmentation had a significant decrease in the patients after the FSH treatment (p<0.05). In addition, the ROS level in sperm, and the malondialdehyde level of seminal plasma significantly decreased after the treatment (P<0.05).ConclusionThe findings indicated that the rhFSH treatment significantly improved the sperm parameters. Further, the treatment led to a meaningful reduction of sperm DNA fragmentation and oxidative stress in the oligozoospermia infertile patients. Similarly, the malondialdehyde concentration markedly decreased in correlation with DNA fragmentation after the rhFSH treatment.
{"title":"The effect of rhFSH on Sperm DNA Fragmentation and sperm parameters in Oligozoospermia Infertile Men","authors":"Atefeh Verdi, S. Sahraei, E. Asa, Rahil Jannatifar, Mohamad Bagher Masaeimanesh","doi":"10.32598/RMM.8.2.1130.1","DOIUrl":"https://doi.org/10.32598/RMM.8.2.1130.1","url":null,"abstract":"BackgroundOne of the main causes of male infertility is the negative effects of oxidative stress. Follicle-stimulating hormone (FSH) plays an essential role in spermatogenesis, as well as in the maintenance of sperm DNA integrity. This study aimed to determine whether the recombinant human follicle-stimulating hormone (rhFSH) treatment of sperm parameters could positively affect sperm DNA and oxidative DNA fragmentation in oligozoospermia infertile men.Materials and MethodsThis interventional study was carried out on a sample of 50 oligozoospermia infertile men. To this end, sperm DNA fragmentation and ROS as an oxidative stress marker were measured before and after treatment with the rhFSH sperm parameters.ResultsThe sperm parameters (concentration, mobility, and morphology) were significantly different in the oligozoospermia infertile patients before and after the rhFSH treatment (p<0.05). Moreover, sperm DNA fragmentation had a significant decrease in the patients after the FSH treatment (p<0.05). In addition, the ROS level in sperm, and the malondialdehyde level of seminal plasma significantly decreased after the treatment (P<0.05).ConclusionThe findings indicated that the rhFSH treatment significantly improved the sperm parameters. Further, the treatment led to a meaningful reduction of sperm DNA fragmentation and oxidative stress in the oligozoospermia infertile patients. Similarly, the malondialdehyde concentration markedly decreased in correlation with DNA fragmentation after the rhFSH treatment.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"55-62"},"PeriodicalIF":0.0,"publicationDate":"2020-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44557477","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}
Behnam Mortazavi, N. Allahyari Fard, F. Heidari, A. Karkhaneh, Mohammad Ali Eslamizade
BackgroundContraceptive vaccines (CVs) can be used as a valuable and alternative method for the prevention of gestation in humans and animals. These vaccines can have several targets, such as superficial sperm proteins. Vaccines based on sperm antigens are quite efficacious to create a contraceptive effect. However, multi-epitope vaccines are more effective in stimulating the immune system and producing more antibodies to reduce the infertility rate.Materials and MethodsThis study aimed to design and evaluate a chimeric fusion protein containing IZUMO, SACA3, and PH-20 epitopes. IZUMO1, SACA3, and PH-20 were assessed, and appropriate regions were selected using various bioinformatics tools, including IEDB, I-TASSER, ProtParam, Asa-View, and Chimera software. Protein epitopes were selected based on various characters, including specificity, solvent accessibility, their weight and length, antigenic intensity, and topology. Epitopes with high antigenic potential were selected and joined together by linkers. The designed fusion protein was simulated using Molecular Dynamic, GROMACS 5, and Chimera 1.14 software.ResultsThe results demonstrated that all antigenic plots and availability of epitopes in the new construct remained constant. The spermatic antigens were combined using rigid linkers as a new construct and showed a stable formation with proper solvent accessibility validated by ProSA-web and PROCHECK. Also, comparing the new structure with its original one did not show any structural change.ConclusionBased on bioinformatics results, the fusion protein that consists of three spermatic antigens has productive potential to stimulate the immune system and capable of producing more antibodies in circulation and reliable infertility.
{"title":"Designing a Construct of Chimeric Multi-Epitopes Protein for Contraceptive Vaccine in Mice: An Immunoinformatics and In Silico Study","authors":"Behnam Mortazavi, N. Allahyari Fard, F. Heidari, A. Karkhaneh, Mohammad Ali Eslamizade","doi":"10.32598/RMM.8.2.894.1","DOIUrl":"https://doi.org/10.32598/RMM.8.2.894.1","url":null,"abstract":"BackgroundContraceptive vaccines (CVs) can be used as a valuable and alternative method for the prevention of gestation in humans and animals. These vaccines can have several targets, such as superficial sperm proteins. Vaccines based on sperm antigens are quite efficacious to create a contraceptive effect. However, multi-epitope vaccines are more effective in stimulating the immune system and producing more antibodies to reduce the infertility rate.Materials and MethodsThis study aimed to design and evaluate a chimeric fusion protein containing IZUMO, SACA3, and PH-20 epitopes. IZUMO1, SACA3, and PH-20 were assessed, and appropriate regions were selected using various bioinformatics tools, including IEDB, I-TASSER, ProtParam, Asa-View, and Chimera software. Protein epitopes were selected based on various characters, including specificity, solvent accessibility, their weight and length, antigenic intensity, and topology. Epitopes with high antigenic potential were selected and joined together by linkers. The designed fusion protein was simulated using Molecular Dynamic, GROMACS 5, and Chimera 1.14 software.ResultsThe results demonstrated that all antigenic plots and availability of epitopes in the new construct remained constant. The spermatic antigens were combined using rigid linkers as a new construct and showed a stable formation with proper solvent accessibility validated by ProSA-web and PROCHECK. Also, comparing the new structure with its original one did not show any structural change.ConclusionBased on bioinformatics results, the fusion protein that consists of three spermatic antigens has productive potential to stimulate the immune system and capable of producing more antibodies in circulation and reliable infertility.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"71-82"},"PeriodicalIF":0.0,"publicationDate":"2020-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45548065","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}
BackgroundCell viability and apoptosis are two crucial factors that may determine cell fate. There are several factors, such as hypoxia, which may be effective in cell processes. Because of its unique features, such as its antioxidant, anti-inflammatory, and anti-apoptosis mechanisms, kohlrabi (Brassica oleracea var. gongylodes) extract may be used in the amelioration of cell viability and a decrease in cell apoptosis. In this study, we evaluate the effect of kohlrabi extract on the viability and apoptosis of adipose-derived mesenchymal stem cells (AD-MSCs).Material and MethodsIn this study, extract from kohlrabi and mesenchymal stem cells from adipose tissue were isolated in a laboratory under sterile conditions. Expression of mesenchymal stem cell (MSC) surface markers, including CD44, CD90, and CD105 was evaluated by flow cytometry method. Besides, CD34 was used as a negative marker. MTT assay was carried out to determine the cell viability. Evaluation of BCL2 and BAX expression levels was performed by real-time PCR.ResultsMSC surface markers were verified by flow cytometry. The obtained results demonstrated a significant difference between the cell viability of the kohlrabi-extract treated and control group over time (P=0.03). In addition, the real-time PCR analysis showed that expression levels of BCL2 significantly increased in hypoxic condition after treatment with leaf extract (P=0.019). However, there was no significant expression change in the BAX gene.ConclusionOur study illustrates that kohlrabi extract may have positive effects on cell survival while having inhibitory effects on apoptosis.
{"title":"Evaluation of Kohlrabi (Brassica oleracea Var. Gongylodes) Extract Effect on Mesenchymal Stem Cells Viability and Apoptosis","authors":"Naser Kalhor Qom, Mohsen Sheykhhasan, A. Kowsari","doi":"10.32598/RMM.8.2.1","DOIUrl":"https://doi.org/10.32598/RMM.8.2.1","url":null,"abstract":"BackgroundCell viability and apoptosis are two crucial factors that may determine cell fate. There are several factors, such as hypoxia, which may be effective in cell processes. Because of its unique features, such as its antioxidant, anti-inflammatory, and anti-apoptosis mechanisms, kohlrabi (Brassica oleracea var. gongylodes) extract may be used in the amelioration of cell viability and a decrease in cell apoptosis. In this study, we evaluate the effect of kohlrabi extract on the viability and apoptosis of adipose-derived mesenchymal stem cells (AD-MSCs).Material and MethodsIn this study, extract from kohlrabi and mesenchymal stem cells from adipose tissue were isolated in a laboratory under sterile conditions. Expression of mesenchymal stem cell (MSC) surface markers, including CD44, CD90, and CD105 was evaluated by flow cytometry method. Besides, CD34 was used as a negative marker. MTT assay was carried out to determine the cell viability. Evaluation of BCL2 and BAX expression levels was performed by real-time PCR.ResultsMSC surface markers were verified by flow cytometry. The obtained results demonstrated a significant difference between the cell viability of the kohlrabi-extract treated and control group over time (P=0.03). In addition, the real-time PCR analysis showed that expression levels of BCL2 significantly increased in hypoxic condition after treatment with leaf extract (P=0.019). However, there was no significant expression change in the BAX gene.ConclusionOur study illustrates that kohlrabi extract may have positive effects on cell survival while having inhibitory effects on apoptosis.","PeriodicalId":30778,"journal":{"name":"Research in Molecular Medicine","volume":"8 1","pages":"93-102"},"PeriodicalIF":0.0,"publicationDate":"2020-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"45136349","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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