Pathobiology Research
Year:2015 | Volume:18 | Issue:3|Papers Count:12

Objectives: Although several studies have examined the association between gene polymorphism GNB3 and endurance exercise, the results are inconsistent. This systematic review and meta-analysis aims to summarize the relationship between GNB3 C825T polymorphism and endurance exercise performance.Methods: We searched all studies published until January31, 2015 in databases PubMed, Google Scholar, Embase, Medline, Science Direct and SID. From ten studies of gene polymorphisms GNB3 and endurance exercise, we selected three studies for the met aanalysis.Results: No significant association existed between GNB3 polymorphism and endurance exercise in the T versus C allelic model (OR: 1.127; 95% CI: 0.877, 1.448; P=0.349),TT versus CC additive model (OR: 1.316; 95% CI: 0.900, 1.924; P=0.157), TT + CT versus CC dominant model (OR: 1.098; 95% CI: 0.856, 1.408; P=0.464), and the TT versus CT + CC recessive model (OR: 0.520; 95% CI: 0.520, 1.111; P=0.157).Conclusion: The results showed that the GNB3 gene polymorphism was not associated with improving the performance of elite athletes in endurance sports. However, further studies would be needed in different ethnicities.

Objective: This study aimed to evaluate the effect of vitrification on the morphology of human ovarian tissue in a stimulated group compared to a non-stimulated group.Methods: Ovarian cortex biopsies collected from stimulated and non-stimulated groups were transported to the laboratory in L-15 medium. Biopsies were cut into small pieces and divided randomly into the vitrified and non-vitrified subgroups. The vitrified-warmed and fresh samples were fixed using Bouin’s solution, then passaged, sectioned and stained with hematoxylin and eosin and Masson’s trichrome. The follicles at different developmental stages were counted and evaluated.Results: Morphological observations showed that the follicles and stromal tissue had well preserved normal structures after vitrification and warming. The percentage of normal follicles in the non-stimulated non-vitrified group was 89.22%; for the non-stimulated vitrified group, it was 84.60%. In previous groups the proportion of primordial follicles were 68.7 ± 1.60% and 67.80 ± 3.71%, primary follicles were 28.60 ± 1.72% and 29.40 ± 3.51%, and secondary follicles were 3.60 ± 0.66% and 2.70 ± 1.20%, respectively. The percentage of normal follicles in the stimulated non-vitrified group was 88.18%; for the stimulated vitrified group, it was 84.19%. In stimulated non-vitrified and stimulated vitrified groups the proportion of primordial follicles were 49.70 ± 4.13% and 49.34 ± 2.86%, primary follicles were 44.50 ± 3.83% and 44.72 ± 2.68%, and secondary follicles were 5.60 ± 0.72% and 5.91 ± 0.77%, respectively. There was no significant difference between the vitrified and non-vitrified and stimulated and non-stimulated groups.Conclusion: Vitrification had no harmful effect on the morphology of stimulated human ovarian tissue and stroma and ovarain tissue structure was similar to the non-stimulated group. This method could be a good alternative for fertility preservation.

Objective: Apical membrane antigen-1 (AMA-1) is one of the most promising blood stage candidate antigens for production of a malaria vaccine against the Plasmodium parasite. Genetic diversity in protective antigens, which is a common phenomenon in a complex pathogen such as the Plasmodium parasite, is responsible for problems with the development of an effective malaria vaccine. This phenomenon will increase the parasite’s ability to evade immune responses. Therefore, malaria vaccine development requires the evaluation of immune responses to different allelic forms of the vaccine candidate antigens.Methods: In this investigation, the two variant forms of PvAMA-1 (PvAMA-1A and B) were expressed in an Escherichia coli M15-pQE30 system using genomic DNA from Iranian individuals with patent Plasmodium vivax infection. The IgG responses of two antigens were evaluated in BALB/c mice with the purified protein emulsified in Freund’s adjuvant. In addition, the correct conformation of the recombinant proteins was evaluated by the indirect immunofluorescence antibody test (IFA).Results: The evaluation of immunogenic responses of two variant forms of PvAMA-1 showed the presence of IgG responses in mice after three immunizations. Cross-reactions were observed. Monitoring of IgG responses showed the persistence up to one year after the last immunization. The antibodies raised against recombinant PvAMA-1s in injected mice recognized the native protein (PvAMA-1) localized on Plasmodium vivax merozoites.Conclusion: The present outcomes confirmed the presence of common epitopes in recombinant forms of the protein that corresponded to native proteins. These emulsified proteins in Freund’s adjuvant were immunogenic in BALB/c mice and IgG responses persisted for up to one year. The IgG responses to two PvAMA-1 variants did not differ significantly. The presence of cross-reactive antibodies has implied that one of these two forms of protein could be used in a universal blood-stage vaccine based on the PvAMA-1 antigen.

Objective: The use of stem cells, particularly mesenchymal stem cells (MSCs), with genes and various growth factors as treatments for myocardial infarction and various other diseases is highly regarded. However these cells meet with inflammation and a hypoxic environment in the target tissue. Hence, treatment with factors that increase the resistance of these stem cells is of importance. Stem cells also can be used as carriers for gene therapy. The aim of the present research is to produce VEGF expressing MSCs. We investigate the effect of stromal derived factor 1 on MSC survival in order to use these cells in a future rat myocardial infarction model.Methods: MSCs were purified from young male rats by aspirating the cavity of femurs and tibias. After characterization, MSCs were transduced with VEGF using lipofectamine. Expression and function of VEGF was confirmed. Next, we treated MSCs with SDF1a at various time points. The effect of this chemokine was investigated using the LDH assay and by viable cell counts.Results: The experiments confirmed the production and function of VEGF by MSCs. The LDH levels decreased significantly in SDF1a treated MSCs. Cell viability increased significantly in the presence of this chemokine.Conclusion: Treatment of MSCs with the SDF1a chemokine has increased the survival of these cells. These MSCs are proper candidates for increasing angiogenesis and for further analysis in a rat model of myocardial infarction.

Objective: The present study compared the efficiency of three mouse ovarian tissue culture methods - hanging drop, under mineral oil, and on the insert with regards to improving in vitro ovarian follicular development.Methods: Ovaries from 7-day-old old female NMRI mice sacrificed by cervical dislocation were collected and cultured in α-MEM medium supplemented with 5% fetal bovine serum for 7 days in 3 groups (hanging drop, under mineral oil and on the insert). We evaluated and compared the morphology and surface area of the ovaries and percentage of normal follicles in all groups. After the 7-day culture, the ovaries cultured on the insert showed better growth compared to the other groups. Their preantral follicles were isolated and cultured for 12 days. We assessed the follicular diameter, survival and maturation rate of these oocytes at the end of the last culture period.Results: The percentages of normal follicles in cultured ovaries were 73.85±2.49% (insert), 51.63±3.93% (hanging drop), and 40.52±5.86% (mineral oil) after the 7-day culture. The percentage of preantral follicles significantly increased from 2.1±0.44 to 24.5±2.4 in the group cultured on insert (P<0.05), however there was no significant difference in the other groups (P>0.05). There were significantly increased surface areas of the ovarian tissues after the 7-day culture in all groups (P<0.05). Ovaries cultured on the insert had a diameter of isolated follicles after 12 days of culture of 410±7.07 μm and the MII rate was 30.26%.Conclusion: The ovarian growth and morphology were well preserved in tissues cultured on the insert compared to the other culture methods.

Objective: Ilam is a border province and a high risk zone for zoonotic cutaneous leishmaniasis (ZCL). Identification of Leishmania parasite species in clinical infections is a prerequisite for planning appropriate control measures. This study investigates the demographic characteristics of patients and molecular epidemiology of Leishmania parasites in the skin lesions of patients from Ilam Province.Methods: A total of 106 cases of suspected cutaneous leishmaniasis were detected passively and microscopic slides prepared from their active skin lesions. We randomly selected 50 slides. A fragment of the rDNA-ITS1 gene was amplified after which the PCR products digested with HaeIII restriction enzyme. There were 18 samples sequenced and their phylogenetic relationships compared with sequences retrieved from GenBank.Results: Leishmania amastigotes were detected in 100 slides. The highest and lowest distribution of cases was from the Moosian and Dehloran districts, respectively. There were 68.9% males and 31.1% of cases were women. The RFLP pattern of all samples was similar to Leishmania major. Phylogenetic relationships displayed great similarity between our sequences and those of Leishmania major parasites from sandflies trapped in Ilam and South Khorasan Provinces and human hosts from Esfarayen, Mahshahr and Afghanistan plus Leishmania mexicana of Venezuelan origin classified together in the same clade.Conclusion: Due to homogeneous morphology, problems associated with the cultivation of Leishmania and the two-step molecular identification process, the rDNA-ITS1-RFLP method has gained considerable attention in recent years. This method could be used as a very sensitive, simple, rapid and inexpensive method to detect Leishmania parasites in a variety of clinical and non-clinical samples.

Objective: Worldwide, Leishmania major is one of the major causes of cutaneous leishmaniasis, including Iran. In the present study we investigate the effect of a direct electricity current in combination with silver nanoparticle on the killing of Leishmania major in vitro.Methods: We evaluated the effects of different concentrations of silver nanoparticles against Leishmania major promastigotes in vitro, then the half maximal inhibitory concentration (IC50) of the nanoparticles was determined. In the second step, the killing effect of silver nanoparticles alone or in combination with 3mA of direct electric current was assessed in promastigote cultures for 10 minutes. Next, we evaluated the survival rate of treated promastigotes with the MTT assay.Results: The parasite count showed that the various concentrations of silver nanoparticles significantly decreased the numbers of live promastigotes over time compared with the control group after 24, 48 and 72 hours of culture. The IC50 of the nanoparticles was 39.8 mg/ml after 48 hours of cultivation. Promastigote mortality occurred in 33.5% with the use of silver nanoparticles alone at concentrations of 160 mg/ml and 100% when combined with 3 mA direct current electricity after 10 minutes.Conclusion: Silver nanoparticles alone did not completely kill Leishmania major promastigotes. However, the combined use of both direct current electricity and silver nanoparticles had a significant synergistic effect on promastigote mortality.

Objective: Azole antifungal drugs have been a treatment option for Candida albicans infections. However, azole resistance can occur through different mechanisms such as alterations in ERG11 (lanosterol 14α-demethylase). This study assesses ERG11 gene mutations in Candida albicans strains isolated from patients with Candidia Volvo vaginitis in a number of Rasht hospitals between 2012-2014 by direct PCR and sequencing.Methods: We identified the yeast strains by standard identification methods, such as germ tubes. Drug sensitivity was determined as MIC 90 values by the macro dilution broth method based on the CLSI protocol. We screened the resistant strains prior to DNA extraction and ERG11 gene mutations were confirmed by PCR sequencing.Results: From 40 strains, 4 showed high levels of resistance to fluconazole. Of these, two species had a MIC 90 of 512mg/ml and the other two species had a MIC 90 of 1024 mg/ml. Three strains had D116E and V456G polymorphisms.Conclusion: The most fluconazole resistant Candida albicans strains worldwide were reported. Our results suggested a correlation between the polymorphism and fluconazole resistance in the Candida albicans strains.

Objective: Interleukin 6 (IL6) is a pro-inflammatory cytokine that plays an important role in obesity and related metabolic disorders. Also, exercise training has been offered in obesity prevention and immune system improvement. Hence, this study intends to survey the effects of high fat diet and high intensity aerobic training on plasma levels of IL6 in rats.Methods: We divided 28 male Wistar rats into two control groups with normal (CN) or high fat (CH) diet and two training groups with normal (EN) or high fat (EH) diet. The training groups ran for 60 minutes on a treadmill at 35 m/min for 5 days/week (75% VO2 max). After 8 weeks, we collected blood samples for plasma IL6 assessment. Two-way ANOVA was used for statistical analysis (P£0.05).Results: The high fat diet significantly increased the rats’ weights and plasma IL6 levels. Performance of 8 weeks of high intensity aerobic training increased IL6 levels in the normal diet group and decreased IL6 levels in the high fat diet group. This change was not significant in the normal diet group.Conclusion: High fat diets probably induced inflammation due to elevated IL6 levels. High intensity aerobic training for 8 weeks significantly decreased IL6 levels.