Pathobiology Research
Year:2020 | Volume:23 | Issue:2|Papers Count:7

Aims Many inhibitors have been introduced for the treatment of HIV-1 infections; however, most of these efforts have been failed due to the presence of resistant strains. The purpose of the current study was to investigate the treatment-resistance mutations in the HIV virus integrase gene and the effect of these mutations on the structure, function, and physical and chemical properties of this enzyme using bioinformatics software. Materials & Methods 36 HIV-1 integrase sequences form Iranian patients were obtained from the NCBI Genbank. After determining the mutations compared to the reference sequence, its post-modification and physical and chemical properties were described. Sequences subtypes, as well as the second and third structures, and possible interactions of this enzyme with the main inhibitors of the integrase were examined. Findings The analysis of selected sequences indicated a number of mutations in this protein. The subtype of most of the samples was A1 and the results of the analysis of the interaction showed that the mutations in the samples had no significant effect on the interaction of inhibitors with the integrase enzyme. Conclusion The binding site of these inhibitors is often found in the catalytic domain of integrase enzyme, and the results of this study depicted that most mutations were located outside this region, and this may be the main reason for the failure of these mutations to affect the interaction of inhibitors and integrase enzyme. Generally, the findings of this study suggest that anti-HIV inhibitors of HIV-1 can be used as an effective way to control this disease for Iranian patients.

Aims The development of new antiviral agents is an appropriate approach to eradicate hepatitis C infection. Due to the lack of suitable animal models, there is always a barrier to the proper evaluation of antiviral compounds in vivo. The growing attention to microRNAs is a new strength in antiviral therapy. The aim of the present study was to use luciferase assay to confirm the specific interaction between miRNA and genomic RNA of hepatitis C virus (HCV) genotype 1b to suppress the replication of the virus. Materials & Methods The NS5B genomic fragments of the HCV genome were sub-cloned into the psiCHECK-2-TM vector as MRE. The relative expression of lentivirus vectors expressing miRNAs in Huh7. 5 cells was assessed through fluorescence microscopy and real-time PCR. The lentivirus expressing let-7b was transduced to Huh7. 5 cells. The NS5B-psiCHECK-2-TM (MRE) was transfected to the Huh7. 5 cell. The relative expression of luciferase was measured using a luciferase dual assay kit. Findings With the use of lentiviruses expressing let-7b, high and permanent expression of let-7b was created in the target cell. On the other hand, the specific attachment of the responsive sequence (NS5B) to the microRNA of let-7b was shown by decreasing luciferase light. Conclusion Lentiviral vectors are used to maintain high and stable expression of microRNAs in cells. The use of luciferase assay is one of the most appropriate methods to confirm the interaction between miRNA-mRNA that can be used for other viral genes with different microRNAs.

The human leukocyte antigen (HLA) system has unique genetic and biochemical complexes, so it is important to determine the type of HLA in a clinical and laboratory process. In addition, to its application to assess the risk of various diseases, it also plays a vital role in organ transplants. The determination of HLA in patients requiring transplantation, in addition to the accuracy, also requires the speed of action. So that any delay and error in this work can endanger the patient’ s life. The need for a precise definition of the HLA type, along with improved speed and cost reduction, has led to the introduction of several methods for this test. Today, the introduction of a new generation of revolutionary sequencing has been carried out in genetic assessments. The HLA determination also uses this technique, and a variety of methods have been introduced for genetic HLA examination using a new generation of sequencing. In the present study, along with the review of traditional methods in HLA diagnosis, two of the most widely used techniques based on a new generation of sequencing methods, Thermal Sequencing method and Illumina Miseq method were evaluated.

Aims Smokers are exposed to significant quantities of oxidative factors. The exercise has been shown to increase activation of antioxidant enzymes and reduce the production of free radicals in the body. Therefore, the present study was investigated the effect of 12 weeks of combined training on oxidative stress and antioxidants capacity in smoker’ s football players. Materials & Methods 22 smoker’ s football players with normal weight and the average age of 23. 9± 1. 9 years were randomly divided into two experimental and control groups. The experimental group submitted to combine training including aerobic and resistance exercise (3 sessions per week) for 12 weeks. Antioxidant indicators (catalase and superoxide dismutase) and lipid peroxidation indicator (malondialdehyde) were measured 48 hours before and after protocol at least 8 hours of fasting. Dependent t-test was used to investigate the differences within the group data, and independent t-test was applied to investigate intergroup differences. The significance level was p≤ 0. 05. Findings 12 weeks of combined training (aerobic and resistance) was caused respectively significant increase and decrease amounts of enzymes CAT and SOD as antioxidant indicators and MDA as lipid peroxidation indicators in smoker’ s football players (p≤ 0. 05). Conclusion Combined exercise training (aerobic and resistance) likely by increase antioxidant capacity and decrease lipid peroxidation indicators eliminates the oxidative stress in smoker’ s football players.

Aims Vitrification affects intracellular calcium, fertilization ability, and developmental competence of mammalian oocytes. This effect may be more closely associated with an intracellular calcium rise induced by cryoprotectants. The present study aimed to assess whether reducing calcium of vitrification solution could improve the fertilization and developmental competence of ovine oocytes. Materials & Methods COCs were collected from the ovine ovary. MII oocytes were divided into 5 groups, one non-vitrified (control) and four vitrified groups 24 hours after COC culture. Vitrified groups were designed according to the presence or absence of EGTA (a calcium chelator) and/or calcium in base media, including mPB1+ (modified PBS with Ca2+), mPB1-(modified PBS without Ca2+), mPB1+/EGTA (mPB1+ containing EGTA), mPB1-/EGTA (mPB1-containing EGTA). Fertilization rate and in vitro development were evaluated after embryo thawing. Also, blastocyst quality was assessed using differential staining. Data analysis was carried out using one-way analysis variance. Findings There was no significant difference in the viability rate between vitrified groups. Fertilization and the developmental rate decreased in the presence of calcium (p<0. 05) but in the calcium-free medium with the EGTA supplementation group, the developmental rate obviously increased. On the other hand, blastocyst cell count in the control group was similar to vitrified groups. Conclusion Using a calcium-free cryoprotectant by adding EGTA can improve the quality of vitrified-thawed ovine MII oocyte and also a higher developmental rate in obtained embryos.

Aims The present study aims to compare the survival rate, reconstitution and dislocation of meiotic spindle of mouse oocyte vitrification between open and close systems. Materials & Methods 131 matured oocytes (containing meiotic spindle) harvested from 6-8 week-old BALB/C and divided into two groups of open and close systems. After exposure of oocytes in vitrification media, oocytes were loaded on cryotop in open group and on Rapid-i in close group, and then vitrified. After warming, oocytes were incubated for 3 hours and evaluated for survival rate, presence and location of re-polymerized meiotic spindle by PolScope system. Findings Survival rate was significantly higher in open system compared to close system (91. 1% vs 68. 6%, respectively). 81. 3% and 51. 2% of survived oocyte contained meiotic spindle in close and open system, respectively (p<0. 05). The percentage of nature location of meiotic spindle was more in close system, significantly. Conclusion Survival rate in open system is more than close system, but close system can preserve meiotic spindle and nature location better than open system. Also, it seems that the evaluation of meiotic spindle is essential for vitrified oocytes by PolScope system.

Aims Despite the efficacy of current therapies against HIV-1 infection, these methods are not a permanent treatment because they cannot prevent the return of viremia from latent cell reservoirs. On the other hand, the virus may become resistant to these drugs. Therefore, providing safer and more effective therapeutic strategies, such as inhibition of genes by siRNA, is essential. The successful therapeutic application of siRNAs requires an efficient delivery system to target cells. Materials & Methods In this study, a specific siRNA was designed against the HIV-1 nef gene. Then a stable HEK293 cell line expressing HIV-1 nef was developed and after fabrication and evaluation of superparamagnetic iron oxide nanoparticles (SPION) coated with trimethyl chitosan, the efficiency of nanoparticles for delivering siRNA into the cells and inhibition of nef gene was investigated. Findings Iron oxide nanoparticles (spherical-shaped with an average size of 85nm and the average zeta potential of +29mV) were significantly effective in transporting siRNA into HEK293 cells compared to control groups and at the same time had low toxicity to the cells. In addition, SPION-TMC containing anti-nef siRNA inhibited about 85% of the expression of this gene in stable cells (compared to control cells). Conclusion The optimized SPION-TMC nanocarriers can be used as a promising approach in HIV-1 infection therapy. However, pre-clinical in vivo evaluation of the drug/siRNA delivery system efficiency remains to be conducted.