Research in Molecular Medicine (RMM)
Molecular medicine is a rapidly expanding field that has revolutionized our understanding of pathophysiology of diseases. This field has grown to encompass researchers with backgrounds in many diverse fields, including pathology, immunology, biochemistry, molecular biology, genetics, microbiology, clinical medicine, and molecular epidemiology. Important contributions to this field have come from work with populations, with patients, with animal models, and with in vitro systems. The application of research involving gene technology, gene therapy, Immunological aspects of diseases, molecular structural analysis, molecular epidemiology and molecular and clinical pharmacology, pathology, and Microbial pathogenesis has made unprecedented progress and precision possible in the understanding, prevention, diagnosis and treatment of human diseases. These areas of molecular medicine, therefore, will be given particular attention by the editorial board of Research in Molecular Medicine (RMM).
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Materials and Methods: In 2020, 27 patients (21 males and 5 females) with imported P. falciparum cases were studied. The nested-PCR technique first confirmed the species in all samples and then amplification was done by the semi-nested-PCR method in order to detect single nucleotide polymorphisms (SNPs) in dhfr gene related to pyrimethamine resistance.
Results: All samples in the 18S rRNA gene had species-specific bands for P. falciparum strains. In the sequence analysis of pfdhfr gene amplification after comparison with the standard strain (wild type), 21 patients had a double mutation (C59R+S108N) and six patients had a triple mutation (N51I+C59R+S108N) of pyrimethamine resistance.
Conclusion: The results of this study showed that the susceptibility of P. falciparum to pyrimethamine in the treatment of malaria is significantly reducing. These findings can raise concerns about pyrimethamine resistance in P. falciparum. Due to the high emergence of double and triple mutants related to pyrimethamine resistance, the malaria surveillance and treatment systems in Iran, the use of pyrimethamine should be considered.
Materials and Methods: In our study, 74 pregnant women diagnosed with GDM according to the American Diabetes Association criteria and 49 healthy pregnant women were included. DNA isolations were made from peripheral blood cells collected from pregnant women and regions of targeted genes were scanned by the Polimerase Chain Reaction-Restriction fragment length polymorphism (PCR-RFLP) technique. The homeostatic model assessment for insulin resistance (HOMA-IR), which is an indicator of insulin resistance, was calculated for each individual in the biochemical examinations. The associations of genotypes detected in the target gene regions with the disease and their effects on the biochemical phenotypes were analyzed by establishing the dominant, recessive, and additive models along with calculating odd ratios. The P<0.05 was considered statistically significant in all analyses.
Results: A statistically significant association was found between R1273R substitution in the ABCC8 gene and GDM under dominant and additive models. No statistically significant correlation was found between the A1369S and e16/-3t→c variants in the ABCC8 gene and the screened variants in other genes and GDM. When the genotype-phenotype association data was evaluated, no association was detected between all the scanned variants and fasting blood sugar while a weak correlation was found between e16/-3t→c in the ABCC8 gene and fasting insulin (P=0.075) and HOMA-IR (P=0.067).
Conclusion: ABCC8 (R1273R and e16/-3t→c) gene variants may be a risk factor for the development of GDM in the Turkish population.
Materials and Methods: This experimental study was conducted on 24 adult male Wistar rats (eight weeks old and weighing 278.26±18.06g), which were randomly assigned to three groups of healthy control (n=8), diabetic control (n=8), and diabetes+aerobic exercise (n=8). The exercise protocol consisted of eight weeks of exercise, three sessions a week, starting with 10 minutes of running at a speed of 10m/s in the first week and ultimately reaching 40 minutes of running at a speed of 18m/s in the eighth week. The changes were analyzed using the one-way analysis of variance and Tukey’s post hoc test.
Results: Significant differences were observed between the groups in terms of body mass (P=0.0001), fasting glucose (P=0.004), serum insulin (P=0.023), and myocardial Ppargc- 1α expression (P=0.031). The post hoc test represented a notable weight decrease in the diabetic control group (P=0.001) and the diabetic exercise group (P=0.001) compared to the healthy control group. The results also showed a significant increase in the glucose level of the diabetic control group compared to the healthy control group (P=0.008) and a notable decrease in the diabetic exercise group’s glucose level in comparison with the diabetic control group (P=0.001). A significant decrease was also observed in the insulin level of the diabetic exercise group compared to the diabetic control group (P=0.034). The results of the post hoc test for Ppargc-1α expression changes showed significantly increased myocardial Ppargc-1α expression in the diabetic exercise group compared to the diabetic control group (P=0.009). No significant change was detected in the expression of SIRT1 (P=0.075).
Conclusion: The findings suggest that exercise positively affects insulin resistance and weight changes by regulating genes related to mitochondrial biogenesis.
Materials and Methods: The study included 351 healthy men (aged 20-40 years) referred to the organization of blood transfusion in Shiraz City, southern Iran. The participants’ temperament (warm, temperate, and cold) was determined using a standard self-reported scale. Then, we performed Polymerase Chain Reaction (PCR) to determine their 5-HTTLPR polymorphism genotype. Multinomial logistic regression was used to evaluate a 95% CI and odds ratios for the association of temperament with the 5-HTTLPR genotypes. Statistical analysis was performed using the SPSS at a significance level of less than 0.05.
Results: Regarding the warm temperament, no association with the 5-HTTLPR genotypes was observed. However, regarding cold temperament, although our data showed no association with the SS, the LS genotype showed some association. With reference to LL, the LS genotype decreased the possibility of coldness rather than temperateness for the temperament (OR=0.471, P=0.040).
Conclusion: Our data revealed the association of temperament with the 5-HTTLPR polymorphism, suggesting that the serotonergic system may influence temperament. Further studies are required to explore the relation of genetic factors with temperament.
Material and methods: Of the total 120 collected samples for this study, every 60 samples were collected from animals and humans from respective laboratories. Total 76 isolates of P. aeruginosa were isolated and identified by morphological and biochemical tests. The presence of virulence factors like oprL and toxA were evaluated by PCR analysis and antimicrobial resistance was assessed by antibiotic susceptibility test (Kirby-Bauer method).
Results: From the total 76, P. aeruginosa isolates obtained from both animal and human isolates, alone presence and coexistence of both toxA and oprL genes in P. aeruginosa isolates; were detected in PCR analysis. PCR analysis results showed in P. aeruginosa isolates, alone distribution of toxA and oprL genes is, 75% and 54.16% in animals, and 84.61% and 80.76% in humans, respectively. The coexistence of both genes was 37.50% and 40.32% in animals and human isolates, along with high antibiotic resistance in most P. aeruginosa isolates.
Conclusion: Therefore, this study suggested PCR analysis can be used for fast and specific detection of oprL and toxA genes in P. aeruginosa. Monitoring of these genes can help to prevent the risk of transmission of multi-drug resistant P. aeruginosa, from animals to humans.
Materials and Methods: For this purpose, the proteins without allergenicity and high antigenicity, as well as conservancy levels from SARS-CoV-2, were chosen for computational epitope mapping. The T-cell epitope mapping process was performed based on the most frequent human leukocyte antigen (HLA) alleles in Iran. The B- and T-cell epitopes were determined based on their allergenicity, antigenicity, and hemolytic potential. Then, the epitopes with acceptable features were subjected to the final construct. The screened epitopes were structured in the final vaccine sequence. The secondary and tertiary structures of the proposed vaccine were predicted, and its affinity to HLA-I, HLA-II, toll-like receptor (TLR)-3, and TLR-4 were evaluated by the molecular docking method. Additionally, possible immune responses against the vaccine were predicted through immune simulation.
Results: The final vaccine construct includes six linear B-cell epitopes, eight HLA-I restricted epitopes, and six HLA-II restricted epitopes. The evaluations confirmed that the proposed vaccine is a 60.3 kDa stable, water-soluble, and high antigenic protein with high affinity to the selected target molecules and could elicit both humoral and cellular responses.
Conclusion: Altogether, the study results suggest that the planned vaccine can be an adequate anti-COVID-19 vaccine candidate for the Iranian population.
Materials and Methods: In the current review, based on relevantly reported cases, possible mechanisms are suggested on the relationship between the anti-platelet factor 4 (anti-PF4) antibody assays, previous exposure to heparin, and the involved mechanisms of post-vaccination thrombocytopenia and thrombotic events, which might help the experts for selecting the appropriate therapeutic measures.
Results: Possibly involved mechanisms in VITTS after ChAdOx1 nCoV-19 vaccination include binding of anti-PF4 antibodies to heparin/PF4 complex or receptor-binding domain (RBD) protein-PF4 complex. Another mechanism could be the binding of anti-RBD antibodies to the RBD protein-PF4 complex. Finally, anti-RBD or anti-PF4 antibodies may bind to the heparin-RBD protein-PF4 complex. The binding of either of the mentioned antibodies to these complexes via the Fc/angiotensin-converting enzyme 2 receptors can cause activation/removal of platelets leading to thrombocytopenia and thrombosis.
Conclusion: The suggested mechanisms in this article provide a relationship between the results of anti-PF4 antibody assays, previous exposure to heparin, and the involved mechanisms of post-vaccination thrombocytopenia and thrombotic events, which might help the experts in selecting the therapeutic measures.
molecular pathways in the inflamed and non-inflamed intestinal mucosa.
Materials and Methods: We obtain GSE83448 gene expression profiles from the Omnibus gene expression database. Also, for the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of Differentially Expressed Gene (DEG) pathways, we used DAVID software. DEGs were detected in the inflamed and non-inflamed intestinal mucosa of CD patients compared to the control group using the GEO2R instrument. Significant modules and hub genes were identified after producing protein-protein interaction (PPIs) networks of DEGs using Cytoscape software.
Results: The 10 specific hub genes of CD, including Matrix Metallopeptidase 2 (MMP2), Cadherin 1 (CDH1), Periostin (POSTN), Collagen type I alpha 2 chain (COL1A2), C-X-C motif chemokine ligand 8 (CXCL8), Collagen type III alpha 1 chain (COL3A1), JUN, Serine Protease Inhibitor clade E member 1 (SERPINE1), Integrin alpha M (ITGAM), and Connective Tissue Growth Factor (CTGF), were used as biomarkers to discriminate between inflamed and non-inflamed intestinal mucosa groups in patients.
Conclusion: These findings could lead to new molecular targets and diagnostic biomarkers for both inflamed and non-inflamed intestinal mucosa in CD patients.
Materials And Methods: A total of 34 transplant recipients comprising 15 Kidney Transplant (KT) recipients and 19 Bone Marrow Transplant (BMT) recipients admitted to the Imam Reza Hospital in Kermanshah Province, Iran, were enrolled in this study. The CMV viral load was quantified by the real-time PCR technique.
Results: The CMV viral load in KT recipients was significantly higher than in BMT recipients (P=0.03), and there was a positive association between the level of virus and the level of cyclosporine in the blood of patients (r=0.51, P=0.02). Besides, CMV viral load was positively correlated with WBC (r=0.32, P=0.04), urea (r=0.47, P=0.002), creatinine (r=0.39, P=0.01), aspartate aminotransferase (r=0.33, P=0.04), and lactate dehydrogenase (r=0.4, P=0.01). Also, it was negatively associated with albumin (r=-0.61, P<0.001), sodium (r=-0.4, P=0.01), and calcium levels (r=-0.46, P=0.003). There were also significant differences between KT and BMT recipients regarding the CMV-related clinical laboratory findings of urea (P=0.02), creatinine (P=0.001), uric acid (P=0.005), direct bilirubin (P=0.04), albumin (P=0.04), platelet (P<0.001), and sodium (P=0.04) levels.
Conclusion: Based on present data, we conclude that despite careful monitoring of patients, infection with CMV is still one of the most important problems associated with organ transplantation, which is directly related to many laboratory findings.
and C chains, several unwanted products are formed after treatment with these enzymes. To overcome this problem, we introduced three thrombin recognition sites into the proinsulin encoding sequence.
Materials and methods: After the design, the modified proinsulin encoding sequence containing the 5′ His-Tag tail and three thrombin recognition sites located between the His-Tag and B chain, B and C chains, and C and A chains, respectively, was synthesized by overlap extension Polymerase Chain Reaction (PCR) using seven specific primers in multiple sequential PCR reactions. The final amplified fragment was cloned in the pGEM-5zf vector by the EcoRV enzyme. After sequencing, the modified proinsulin encoding sequence was subcloned into the pET-26b(+) vector using NdeI and XhoI enzymes. Finally, the modified proinsulin was expressed in E. coli BL-21(DE3) by induction with Isopropyl β-d-1-thiogalactopyranoside (IPTG).
Results: The accuracy of the synthesized modified proinsulin sequence was confirmed by DNA sequencing. The modified proinsulin cloning was evaluated by PCR with specific primers and digestion with specific restriction enzymes. In this study, the modified proinsulin protein was expressed up to 40%. The modified proinsulin protein expression was assessed using sodium dodecyl Sulfate–Polyacrylamide Gel Electrophoresis (SDS-PAGE) and western blotting.
Conclusions: This modified proinsulin can be used to easily and efficiently produce insulin glargine without any impurities after processing with thrombin in one step in a nickel chromatographic column.
Materials and Methods: A total of 21 male Wistar rats (Mean±SD weight: 220±20 g) were obese by 6 weeks High-Fat Diet (HFD) and randomly assigned to 1) non-diabetic, 2) control T2D, and 3) exercise diabetic groups. T2D was induced by intraperitoneal injection of streptozotocin (30 mg/kg) for diabetic groups. The exercise group did the resistance exercise program (5 times per week for 6 weeks). After exercise training, PI3K and mTORc1 expression in the left ventricle and the ratio of the left ventricle to heart and heart to body weight were compared between groups. The obtained data were compared by 1-way Analysis of Variance (ANOVA) (P<0.05).
Results: Induction of diabetes resulted in significant decrease in all mentioned variables in control diabetic to non-diabetic rats (PI3K; P=0.021, mTORc1; P=0.004, left ventricle/heart weight; P=0.045, heart/body weight; P=0.035). Significant increase was observed in all variables (PI3K; P=0.028, mTORc1; P=0.015, left ventricular/heart weight; P=0.002, heart/body weight; P=0.001) in response to resistance training compared to the control rat.
Conclusion: Based on our results, cardiac hypertrophy in studied diabetic rats can be attributed to improved PI3K/mTORc1 signaling in response to resistance training. Exploring the exact mechanisms responsible for these changes in response to exercise requires further molecular-cellular studies.
Materials and Methods: T2D was induced by HFD/low-dose STZ in 30 male Wistar rats (21-monthold, Mean±SD weight 418±43 g). The rats received HFD (55%, 31%, and 14% of energy from fat, carbohydrate, and protein, respectively; 5.2 kcal/g). The diets continued for eight weeks in both groups. Over week four, the rats in the group with HFD/STZ-induced T2D received treatment with low-dose STZ. After one week of familiarity with the laboratory environment, they were randomly divided into three groups: Diabetic Endurance Training (DET, n=10), Diabetic Resistance Training (DRT, n=10), and Diabetic Control (DC, n=10). The eight weeks of endurance training protocol comprised five sessions of moderate-intensity training (60%-75% velocity at maximal oxygen uptake (vVO2max) and low intensity (30%-30% vVO2max). In 60% Maximum Voluntary Carrying Capacity (MVCC), the resistance group climbed the ladder 14-20 times with 1-minute rest, five days a week.
Results: The results of the 1-way ANOVA test showed no significant change in serum miR-133 expression (P=0.411) and muscle tissue (P=0.077) following resistance and endurance training. However, significant differences were observed in bone marrow miR-133 expression (P=0.003) and Runx2 gene expression (P=0.002) between groups. Tukey’s post hoc tests showed that the bone marrow miR-133 expression had a significant increase following eight weeks of resistance training compared to the endurance training (P=0.006) and control (P=0.002) groups, and bone marrow Runx2 gene expression in rats exposed to resistance training compared to the endurance training (P=0.044) and the control (P=0.018) groups.
Conclusion: It seems that longer periods of exercise are required for cellular changes in the metabolism of these tissues after these exercise protocols. This topic should be studied in future research.
Materials and Methods: In 2020, 27 patients (21 males and 5 females) with imported P. falciparum cases were studied. The nested-PCR technique first confirmed the species in all samples and then amplification was done by the semi-nested-PCR method in order to detect single nucleotide polymorphisms (SNPs) in dhfr gene related to pyrimethamine resistance.
Results: All samples in the 18S rRNA gene had species-specific bands for P. falciparum strains. In the sequence analysis of pfdhfr gene amplification after comparison with the standard strain (wild type), 21 patients had a double mutation (C59R+S108N) and six patients had a triple mutation (N51I+C59R+S108N) of pyrimethamine resistance.
Conclusion: The results of this study showed that the susceptibility of P. falciparum to pyrimethamine in the treatment of malaria is significantly reducing. These findings can raise concerns about pyrimethamine resistance in P. falciparum. Due to the high emergence of double and triple mutants related to pyrimethamine resistance, the malaria surveillance and treatment systems in Iran, the use of pyrimethamine should be considered.
Materials and Methods: In our study, 74 pregnant women diagnosed with GDM according to the American Diabetes Association criteria and 49 healthy pregnant women were included. DNA isolations were made from peripheral blood cells collected from pregnant women and regions of targeted genes were scanned by the Polimerase Chain Reaction-Restriction fragment length polymorphism (PCR-RFLP) technique. The homeostatic model assessment for insulin resistance (HOMA-IR), which is an indicator of insulin resistance, was calculated for each individual in the biochemical examinations. The associations of genotypes detected in the target gene regions with the disease and their effects on the biochemical phenotypes were analyzed by establishing the dominant, recessive, and additive models along with calculating odd ratios. The P<0.05 was considered statistically significant in all analyses.
Results: A statistically significant association was found between R1273R substitution in the ABCC8 gene and GDM under dominant and additive models. No statistically significant correlation was found between the A1369S and e16/-3t→c variants in the ABCC8 gene and the screened variants in other genes and GDM. When the genotype-phenotype association data was evaluated, no association was detected between all the scanned variants and fasting blood sugar while a weak correlation was found between e16/-3t→c in the ABCC8 gene and fasting insulin (P=0.075) and HOMA-IR (P=0.067).
Conclusion: ABCC8 (R1273R and e16/-3t→c) gene variants may be a risk factor for the development of GDM in the Turkish population.
Materials and Methods: This experimental study was conducted on 24 adult male Wistar rats (eight weeks old and weighing 278.26±18.06g), which were randomly assigned to three groups of healthy control (n=8), diabetic control (n=8), and diabetes+aerobic exercise (n=8). The exercise protocol consisted of eight weeks of exercise, three sessions a week, starting with 10 minutes of running at a speed of 10m/s in the first week and ultimately reaching 40 minutes of running at a speed of 18m/s in the eighth week. The changes were analyzed using the one-way analysis of variance and Tukey’s post hoc test.
Results: Significant differences were observed between the groups in terms of body mass (P=0.0001), fasting glucose (P=0.004), serum insulin (P=0.023), and myocardial Ppargc- 1α expression (P=0.031). The post hoc test represented a notable weight decrease in the diabetic control group (P=0.001) and the diabetic exercise group (P=0.001) compared to the healthy control group. The results also showed a significant increase in the glucose level of the diabetic control group compared to the healthy control group (P=0.008) and a notable decrease in the diabetic exercise group’s glucose level in comparison with the diabetic control group (P=0.001). A significant decrease was also observed in the insulin level of the diabetic exercise group compared to the diabetic control group (P=0.034). The results of the post hoc test for Ppargc-1α expression changes showed significantly increased myocardial Ppargc-1α expression in the diabetic exercise group compared to the diabetic control group (P=0.009). No significant change was detected in the expression of SIRT1 (P=0.075).
Conclusion: The findings suggest that exercise positively affects insulin resistance and weight changes by regulating genes related to mitochondrial biogenesis.
Materials and Methods: The study included 351 healthy men (aged 20-40 years) referred to the organization of blood transfusion in Shiraz City, southern Iran. The participants’ temperament (warm, temperate, and cold) was determined using a standard self-reported scale. Then, we performed Polymerase Chain Reaction (PCR) to determine their 5-HTTLPR polymorphism genotype. Multinomial logistic regression was used to evaluate a 95% CI and odds ratios for the association of temperament with the 5-HTTLPR genotypes. Statistical analysis was performed using the SPSS at a significance level of less than 0.05.
Results: Regarding the warm temperament, no association with the 5-HTTLPR genotypes was observed. However, regarding cold temperament, although our data showed no association with the SS, the LS genotype showed some association. With reference to LL, the LS genotype decreased the possibility of coldness rather than temperateness for the temperament (OR=0.471, P=0.040).
Conclusion: Our data revealed the association of temperament with the 5-HTTLPR polymorphism, suggesting that the serotonergic system may influence temperament. Further studies are required to explore the relation of genetic factors with temperament.
Material and methods: Of the total 120 collected samples for this study, every 60 samples were collected from animals and humans from respective laboratories. Total 76 isolates of P. aeruginosa were isolated and identified by morphological and biochemical tests. The presence of virulence factors like oprL and toxA were evaluated by PCR analysis and antimicrobial resistance was assessed by antibiotic susceptibility test (Kirby-Bauer method).
Results: From the total 76, P. aeruginosa isolates obtained from both animal and human isolates, alone presence and coexistence of both toxA and oprL genes in P. aeruginosa isolates; were detected in PCR analysis. PCR analysis results showed in P. aeruginosa isolates, alone distribution of toxA and oprL genes is, 75% and 54.16% in animals, and 84.61% and 80.76% in humans, respectively. The coexistence of both genes was 37.50% and 40.32% in animals and human isolates, along with high antibiotic resistance in most P. aeruginosa isolates.
Conclusion: Therefore, this study suggested PCR analysis can be used for fast and specific detection of oprL and toxA genes in P. aeruginosa. Monitoring of these genes can help to prevent the risk of transmission of multi-drug resistant P. aeruginosa, from animals to humans.
Materials and Methods: For this purpose, the proteins without allergenicity and high antigenicity, as well as conservancy levels from SARS-CoV-2, were chosen for computational epitope mapping. The T-cell epitope mapping process was performed based on the most frequent human leukocyte antigen (HLA) alleles in Iran. The B- and T-cell epitopes were determined based on their allergenicity, antigenicity, and hemolytic potential. Then, the epitopes with acceptable features were subjected to the final construct. The screened epitopes were structured in the final vaccine sequence. The secondary and tertiary structures of the proposed vaccine were predicted, and its affinity to HLA-I, HLA-II, toll-like receptor (TLR)-3, and TLR-4 were evaluated by the molecular docking method. Additionally, possible immune responses against the vaccine were predicted through immune simulation.
Results: The final vaccine construct includes six linear B-cell epitopes, eight HLA-I restricted epitopes, and six HLA-II restricted epitopes. The evaluations confirmed that the proposed vaccine is a 60.3 kDa stable, water-soluble, and high antigenic protein with high affinity to the selected target molecules and could elicit both humoral and cellular responses.
Conclusion: Altogether, the study results suggest that the planned vaccine can be an adequate anti-COVID-19 vaccine candidate for the Iranian population.
Materials and Methods: In the current review, based on relevantly reported cases, possible mechanisms are suggested on the relationship between the anti-platelet factor 4 (anti-PF4) antibody assays, previous exposure to heparin, and the involved mechanisms of post-vaccination thrombocytopenia and thrombotic events, which might help the experts for selecting the appropriate therapeutic measures.
Results: Possibly involved mechanisms in VITTS after ChAdOx1 nCoV-19 vaccination include binding of anti-PF4 antibodies to heparin/PF4 complex or receptor-binding domain (RBD) protein-PF4 complex. Another mechanism could be the binding of anti-RBD antibodies to the RBD protein-PF4 complex. Finally, anti-RBD or anti-PF4 antibodies may bind to the heparin-RBD protein-PF4 complex. The binding of either of the mentioned antibodies to these complexes via the Fc/angiotensin-converting enzyme 2 receptors can cause activation/removal of platelets leading to thrombocytopenia and thrombosis.
Conclusion: The suggested mechanisms in this article provide a relationship between the results of anti-PF4 antibody assays, previous exposure to heparin, and the involved mechanisms of post-vaccination thrombocytopenia and thrombotic events, which might help the experts in selecting the therapeutic measures.
molecular pathways in the inflamed and non-inflamed intestinal mucosa.
Materials and Methods: We obtain GSE83448 gene expression profiles from the Omnibus gene expression database. Also, for the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of Differentially Expressed Gene (DEG) pathways, we used DAVID software. DEGs were detected in the inflamed and non-inflamed intestinal mucosa of CD patients compared to the control group using the GEO2R instrument. Significant modules and hub genes were identified after producing protein-protein interaction (PPIs) networks of DEGs using Cytoscape software.
Results: The 10 specific hub genes of CD, including Matrix Metallopeptidase 2 (MMP2), Cadherin 1 (CDH1), Periostin (POSTN), Collagen type I alpha 2 chain (COL1A2), C-X-C motif chemokine ligand 8 (CXCL8), Collagen type III alpha 1 chain (COL3A1), JUN, Serine Protease Inhibitor clade E member 1 (SERPINE1), Integrin alpha M (ITGAM), and Connective Tissue Growth Factor (CTGF), were used as biomarkers to discriminate between inflamed and non-inflamed intestinal mucosa groups in patients.
Conclusion: These findings could lead to new molecular targets and diagnostic biomarkers for both inflamed and non-inflamed intestinal mucosa in CD patients.
Materials And Methods: A total of 34 transplant recipients comprising 15 Kidney Transplant (KT) recipients and 19 Bone Marrow Transplant (BMT) recipients admitted to the Imam Reza Hospital in Kermanshah Province, Iran, were enrolled in this study. The CMV viral load was quantified by the real-time PCR technique.
Results: The CMV viral load in KT recipients was significantly higher than in BMT recipients (P=0.03), and there was a positive association between the level of virus and the level of cyclosporine in the blood of patients (r=0.51, P=0.02). Besides, CMV viral load was positively correlated with WBC (r=0.32, P=0.04), urea (r=0.47, P=0.002), creatinine (r=0.39, P=0.01), aspartate aminotransferase (r=0.33, P=0.04), and lactate dehydrogenase (r=0.4, P=0.01). Also, it was negatively associated with albumin (r=-0.61, P<0.001), sodium (r=-0.4, P=0.01), and calcium levels (r=-0.46, P=0.003). There were also significant differences between KT and BMT recipients regarding the CMV-related clinical laboratory findings of urea (P=0.02), creatinine (P=0.001), uric acid (P=0.005), direct bilirubin (P=0.04), albumin (P=0.04), platelet (P<0.001), and sodium (P=0.04) levels.
Conclusion: Based on present data, we conclude that despite careful monitoring of patients, infection with CMV is still one of the most important problems associated with organ transplantation, which is directly related to many laboratory findings.
and C chains, several unwanted products are formed after treatment with these enzymes. To overcome this problem, we introduced three thrombin recognition sites into the proinsulin encoding sequence.
Materials and methods: After the design, the modified proinsulin encoding sequence containing the 5′ His-Tag tail and three thrombin recognition sites located between the His-Tag and B chain, B and C chains, and C and A chains, respectively, was synthesized by overlap extension Polymerase Chain Reaction (PCR) using seven specific primers in multiple sequential PCR reactions. The final amplified fragment was cloned in the pGEM-5zf vector by the EcoRV enzyme. After sequencing, the modified proinsulin encoding sequence was subcloned into the pET-26b(+) vector using NdeI and XhoI enzymes. Finally, the modified proinsulin was expressed in E. coli BL-21(DE3) by induction with Isopropyl β-d-1-thiogalactopyranoside (IPTG).
Results: The accuracy of the synthesized modified proinsulin sequence was confirmed by DNA sequencing. The modified proinsulin cloning was evaluated by PCR with specific primers and digestion with specific restriction enzymes. In this study, the modified proinsulin protein was expressed up to 40%. The modified proinsulin protein expression was assessed using sodium dodecyl Sulfate–Polyacrylamide Gel Electrophoresis (SDS-PAGE) and western blotting.
Conclusions: This modified proinsulin can be used to easily and efficiently produce insulin glargine without any impurities after processing with thrombin in one step in a nickel chromatographic column.
Materials and Methods: A total of 21 male Wistar rats (Mean±SD weight: 220±20 g) were obese by 6 weeks High-Fat Diet (HFD) and randomly assigned to 1) non-diabetic, 2) control T2D, and 3) exercise diabetic groups. T2D was induced by intraperitoneal injection of streptozotocin (30 mg/kg) for diabetic groups. The exercise group did the resistance exercise program (5 times per week for 6 weeks). After exercise training, PI3K and mTORc1 expression in the left ventricle and the ratio of the left ventricle to heart and heart to body weight were compared between groups. The obtained data were compared by 1-way Analysis of Variance (ANOVA) (P<0.05).
Results: Induction of diabetes resulted in significant decrease in all mentioned variables in control diabetic to non-diabetic rats (PI3K; P=0.021, mTORc1; P=0.004, left ventricle/heart weight; P=0.045, heart/body weight; P=0.035). Significant increase was observed in all variables (PI3K; P=0.028, mTORc1; P=0.015, left ventricular/heart weight; P=0.002, heart/body weight; P=0.001) in response to resistance training compared to the control rat.
Conclusion: Based on our results, cardiac hypertrophy in studied diabetic rats can be attributed to improved PI3K/mTORc1 signaling in response to resistance training. Exploring the exact mechanisms responsible for these changes in response to exercise requires further molecular-cellular studies.
Materials and Methods: T2D was induced by HFD/low-dose STZ in 30 male Wistar rats (21-monthold, Mean±SD weight 418±43 g). The rats received HFD (55%, 31%, and 14% of energy from fat, carbohydrate, and protein, respectively; 5.2 kcal/g). The diets continued for eight weeks in both groups. Over week four, the rats in the group with HFD/STZ-induced T2D received treatment with low-dose STZ. After one week of familiarity with the laboratory environment, they were randomly divided into three groups: Diabetic Endurance Training (DET, n=10), Diabetic Resistance Training (DRT, n=10), and Diabetic Control (DC, n=10). The eight weeks of endurance training protocol comprised five sessions of moderate-intensity training (60%-75% velocity at maximal oxygen uptake (vVO2max) and low intensity (30%-30% vVO2max). In 60% Maximum Voluntary Carrying Capacity (MVCC), the resistance group climbed the ladder 14-20 times with 1-minute rest, five days a week.
Results: The results of the 1-way ANOVA test showed no significant change in serum miR-133 expression (P=0.411) and muscle tissue (P=0.077) following resistance and endurance training. However, significant differences were observed in bone marrow miR-133 expression (P=0.003) and Runx2 gene expression (P=0.002) between groups. Tukey’s post hoc tests showed that the bone marrow miR-133 expression had a significant increase following eight weeks of resistance training compared to the endurance training (P=0.006) and control (P=0.002) groups, and bone marrow Runx2 gene expression in rats exposed to resistance training compared to the endurance training (P=0.044) and the control (P=0.018) groups.
Conclusion: It seems that longer periods of exercise are required for cellular changes in the metabolism of these tissues after these exercise protocols. This topic should be studied in future research.