Background: There have been reports showing the protective role of inducible Heat-shock protein (HSP) 70 in gastric epithelial cells. HSP70-2 gene is located in the class-III region of the MHC on the short arm of chromosome 6. The HSP70-2 gene has a pst1 site due to an A to G transition at the 1267 position and different genotypes of the HSP70-2 gene have been shown to be associated with a different level of HSP70 mRNA expression. This study was performed to investigate relations between polymorphism of the HSP70-2 gene and risk of peptic ulcer diseases.

Materials and Methods: In this descriptive analytical study, the studied population comprised 100 subjects, attending the Endoscopy Center of Hafez Hospital in Shiraz on 2012. All subjects underwent upper gastroscopy. Genomic DNA was extracted from bioptic tissues. Genotypes were determined in patients and controls using PCR-RFLP.

Results: In the non-ulcer subjects, the HSP70-2 genotype distribution was 20 AA (40%), 26 AG (52%), and 4 GG (8%). Meanwhile, the HSP70-2 genotype distribution in peptic ulcer patients were 5 AA (10%), 44 AG (88%) and 1 GG (2%). The results showed that 1267G/A HSP70-2 Gene polymorphism is associated with peptic ulcer.

Conclusion: It appears that polymorphism of HSP70-2 gene is associated with the susceptibility to peptic ulcer diseases. The analysis showed that the AG genotype increased the risk of peptic ulcer (OR=6.76, 95%CI=2.26-20.20, p=0.0006).

Background: Prion diseases are neurodegenerative disorders which ultimately results in the death of their victims. They are caused by structural transformation of cellular prion (PrPC) to its &beta-rich and anomalous isoform (PrPSc) and the accumulation of amyloid fibrillar deposits in the central nervous system. The precise mechanism underling this conversion is yet to be well understood. This study aimed to investigate the effect of non physiological temperatures on the misfolding mechanism of the human prion protein.

Materials and Methods: The crystal structure of human prion protein (90-231), (PDB code: 2Lej) in pdb format was used as a starting structure in this study. Three model structures of this coordinate structure were used separately to simulate PrPC at 27 , 37 and 47 . Molecular dynamic simulations were then performed using double-precision MPI version of GROMACS 4.5.5 for 10 ns and the results were analyzed using SPSS software, SPDBV and VebLab programs.

Results: The change of temperature from 37 to 27 or 47 induced significant structural changes to PrPC. These tempratures caused PrPC to attain a more folded and less flexible tertiary structure compared to its native structure at 37 . They, also, reduce protein-solvent hydrogen bonds and therefore increasing access of hydrophobic solvent to PrPC which may be behind the lower water solubility of PrPC and its increased resistance to proteolytic degradations.

Conclusion: This study shows that changes of temperatures accelerate structural changes of PrPC and reduce its solubility while rendering it vulnerable to transition into PrPSc.

Background: Regarding the importance of prevention of cardiovascular disease during menopause the purpose of present study was evaluating the influence of eight weeks progressive aquatic exercise on serum Apoproteins of A and B and lipoproteins in obese and normal weight menopause women.

Materials and Methods: This was a semi-experimental study in which 29 menopause women from Isfahan (age: 57.4 ±4.68 years) voluntarily participated in it. According to body mass index (BMI), participants were divided to obese (n=15, BMI= 30.21 ±3.90) or normal weight (n=14, BMI=22.44 ±2.25) groups. Subjects of both groups participated in 8 weeks aquatic exercise, three times a week, by progressive intensity of 50 to 70% of maximal heart rate and duration of 45 minutes.

Results: Findings of study indicated a significant influence of exercise on VLDL, Apo A and B and ApoB/ApoA in in obese group. In normal weight group, exercise caused significant changes in LDL, VLDL, Apo A and B and ApoB/ApoA, but there was no significaut difference between groups in none of study variables.

Conclusion: According to our findings, progressive aquatic aerobic exercise induces similar reduction of some cardiovascular risk factors especially ApoB and ApoB/ApoA and increasing ApoA in obese and normal weight menopause women.

Background: Delivery of antigens directly to dendritic cells enhances the immune responses to the antigen and is an attractive approach for eliciting cellular immune responses against mutagenic pathogens like HIV virus. So the aim of this study is evaluation of immune responses elicited by delivered multi-epitopic HIV-1 tat/pol/gag/env recombinant protein to dendritic cells in sito using &alphaDEC-205 mAb.

Materials and Methods: In this study, recombinant protein expressed by pET23a-HIVtop4 plasmid including HIVtop4 sequence (Gag158-186, Pol150-190, ENV296-323, ENV577-610, Tat1-20 and Tat44-61) in BL21 E. coli cells was used as vaccine model. To exploiting dendritic cells, properties for immunization purposes, we conjugated this recombinant protein chemically to anti body against DEC-205 receptor on these cells. Balb/c mice immunized subcutaneously (s.c.) with conjugated multi-epitopic protein or un-conjugated one (as control) simultaneously with Poly I: C as dendritic cell maturation factor. Lymphocyte proliferation was measured by bromo di uridine assay, Cytotoxicity by Grenzyme B production activity, IL-4, IL-17, IFN- cytokines production and total antibody by direct and indirect ELISA methods in order.

Results: Immunization by anti DEC-205 conjugated peptide led to a significant increase in the proliferative responses of lymphocytes, production of Gr-B, IFN-&gamma, IL-4 and IL-17 cytokines and total antibody titer in comparison with the none targeted groups.

Conclusion: It is concluded that targeting of protein antigens to DEC-205+dendritic cells significantly enhances immune responses in compare to non-targeting strategies.

Background: The current study aims to investigate the impact of high intensity interval training (HIIT) on serum levels of CK and LDH as the muscle damage indicators and on Gelatinase-A (MMP-2) serum levels as the tissae inflammatory marker among young sedentary girls.

Materials and Methods: For this quasi-experimental study, 14 sedentary female college students were selected and randomly divided into two groups including the exercise HIIT group (means and standard deviations of age: 21.28 ± 2.56 (years)  weight:52.86 ±4.95 (kg) and height: 163.1±3.7 (cm)) and the control group (means and standard deviations of age: 20.25 ±7.50 (years) weight:52.64 ±3.67 (kg) and height: 162.4±4.5 (cm)). The experimental group performed six repetitions of one-minute runs at 90%- 95% of HRmax. The blood samples were collected before and 30 minutes after the exercise protocol. The serum CK, LDH and MMP-2 levels were measured using corresponding kits. The data were analyzed through the independent t-test at the significance level of 0.05 (p<0.05).

Results: After collecting and analyzing Data, the results showed that CK and LDH levels increased significantly after performing HIIT, while there was no significant change in MMP-2 due to the HIIT.

Conclusion: It can be concluded that the HIIT protocol will lead to an increase in some indicators of muscle damage such as CK, LDH, and that no significant changes could be observed for MMP-2 as the body's inflammation response.

Background: Newcastle disease virus (NDV) is one of the major pathogens in poultry and vaccination is intended to control the disease, as an effective solution, yet. Fusion protein (F) on surface of NDV, has a fundamental role in virus pathogenicity and can induce protective immunity, alone. With this background, here our aim was to construct a baculovirus derived recombinant bacmid shuttle vector (encoding F-protein) in order to express it in insect cell line.

Materials and Methods: In this experimental study, at first complete F gene from avirulent strain La Sota of NDV was amplified by RT-PCR to produce F cDNA. The amplicon was cloned into T/A cloning vector and afterwards into pFastBac Dual donor plasmid. After the verification of cloning process by two methods, PCR and enzymatic digestion analysis, the accuracy of F gene sequence was confirmed by sequencing. Finally, F-containing recombinant bacmid was subsequently generated in DH10Bac cell and the construct production was confirmed by a special PCR panel, using F specific primers and M13 universal primers.

Results: Analysis of confirmatory tests showed that the recombinant bacmid, expressing of F-protein gene in correct sequence and framework, has been constructed successfully.

Conclusion: The product of this F-containing recombinant bacmid, in addition to its independent application in the induction of protective immunity, can be used with the other individual recombinant baculoviruses, expressing HN and NP genes to produce NDV-VLPs in insect cell line.

Background: Hypertrophic cardiomyopathy (HCM) is a various collection of heart diseases with autosomal dominant inheritance affecting 0.2% of the global population. HCM is also the most common cause of sudden cardiac death in individuals younger than 35 years old. Approximately, 40% of affected cases are associated with MYBPC3 gene. The aim of this study is to investigate the possible presence of mutation in 15 and 18 exons of MYBPC3 gene in patients with HCM in Chaharmahal Va Bakhtiyari province.

Materials and Methods: In this experimental study, 30 HCM patients were selected. DNA was extracted using standard phenol-chloroform method. Certain exons were amplified by PCR method. And then, SSCP and HA methods were run.

Results: Significant differences were observed between the positive control samples and other samples. However, there were no difference in studied exons or shift in the bands.

Conclusion: Mutations in the exons of MYBPC3 gene may cause the HCM disease, and change in other exons may be the causative agent in this geographical region and change in this studied exons may not have contributed to the HCM disease. However, it is necessary to study more patients for getting a better conclusion.

  Background: In spite of designing and applying an effective vaccine against Hepatitis B virus (HBV), chronic infection with this virus is still one of the most important health problems worldwide. Host genetic background including single nucleotide polymorphisms play a significant role in chronicity or clearance of the infection. The final product of programmed cell death 1 gene (PDCD1) is expressed frequently on T-cells and in chronic viral infections, prevent the virus-specific T-cell response against the virus. In this study, the association of a single nucleotide polymorphism (+7146A/G) in intron 4 of PD1 gene with chronic hepatitis B infection in Iranian population has been assessed.

  Materials and Methods: 212 chronic HBV patients and 208 healthy controls were analyzed in this case-control study. Genomic DNA of the studied individuals was extracted and after performing polymerase chain reaction (PCR), polymorphism of +7146 was determined via RFLP method.

  Results: Frequencies of GG, GA and AA genotypes on position 7146 of the intron 4 of PD1 gene were 77.4%, 20.7% and 1.9% in patient group and 80.8%, 15.4% and 3.8% in control group, respectively. After statistical analysis, No significant difference was observed between patient and control groups (p=0.198).

  Conclusion: Genotype frequencies in the studied population are in accordance with the results of previous studies. Results of the present study suggest that there is not any association between A/G single nucleotide polymorphism in intron 4 of PD1 gene and susceptibility to chronic hepatitis B in Iranian population.

Background: Inflammation plays an important role in the pathogenesis of atherosclerosis and change in lifestyle represents a successful strategy to prevent cardiovascular disease. Therefore, the present study investigated the combined effects of regulae Pilates training and Apium Graveolens seed supplement on monocyte chemoattractant protein-1(MCP-1) and C-reactive protein (CRP) in sedentary women.

Materials and Methods: In this quasi-experimental study, 28 healthy sedentary woman with age average 31±5.5 years old are selected in a convenience sampling way and randomely divided into and control, exercise, supplement and exercise+supplement groups. Pilates exercise was consisted of exercise movements at 50% to 80% maximal heart rate in, 3 sessions per week for 8 weeks. Supplement and exercise+supplement groups consumed 1.3 gr Apium Graveolens seed capsule three times a day after meals. Fasting blood samples were collected before and 48 hours after the last interventions. MCP-1 and CRP levels were measured by ELISA method.

Results: Eight weeks of Pilates exercise, supplementation and the combined intervention were associated with a significant increase in MCP-1 and CRP levels (p<0.05). Furthermore Pilates exercise and combined intervention were associated with significantly greater increases in percent changes of the MCP-1 and CRP compared to supplement group (p<0.05).

Conclusion: These findings suggest that the protective effect of Pilates exercise and Apium Graveolens seed supplementation non-drug interventions might in part be due to suppression of the inflammatory processes.

Background: The Amyloid beta (Aβ) level increases in the brain of patients with Alzheimer's disease. The present study aimed to investigate the effects of eight weeks continuous training with low and high intensities on Aβ_1-42 levels in hippocampus of Alzheimer model rats.

Materials and Methods: Fifty male Wistar rats (12 weeks old and mean weight 219.82±13.10 g) were divided into five groups including: healthy control, Alzheimer’s control, Alzheimer's low-intensity training, Alzheimer's high-intensity training and sham. To induce Alzheimer's disease, homocysteine is infused into the rats cerebroventricular (dose of 0.6M). Low intensity groups trained with 20m/min (50-55% VO₂max) and high-intensity groups trained with 27m/min (75-80% VO₂max), 60min/day, and five days per week on the treadmill. For data analysis, one-way ANOVA and post hoc Tukey test were performed (p<0.05).

Results: The Aβ_1-42 levels in hippocampus of Alzheimer's control group was significantly higher than healthy control group (p=0.001) and in training groups with both low and high intensity was significantly lower than Alzheimer's control group (p=0.02). But no significant differences were found between two intensity (p=0.99).

Conclusion: It seems that continuous exercise training, through reducing the level of Aβ_1-42 in hippocampus, can be useful for Alzheimer’s disease model rats and continuous training can be studied as a complementary therapy in Alzheimer's disease.

Background: The evidence suggests that obesity causes the chronic inflammation. Chemerin is a new adipokine which is associated with obesity and metabolic syndrome. The effects of combined training on levels of inflammatory markers specialy chemerin and serum amyloid-A has been less studied.The present study aims to examine the effect of six weeks combined training on plasma levels of chemerin, CRP and SAA and plasma lipid in obese men.

Materials and Methods: 18 obese men were divided into control and experimental groups. General characteristics of subjects serum levels of chemerin, CRP and SAA were examined (by ELISA method), before and after one session training. Endurance training protocol was performed on the large muscles included in 6 weeks running around the track with 60 to 75% HRmax, 4 sessions per week, intense resistance training for 25 to 30 minutes with 50 to 70% of one repetition maximum of 6 stations (2 sets, 12 Reps). Data analysis was performed by T-independent test for comparison of two control and training groups, T-paired test for comparison of two groups before and after the test and significant level of p≤0.05 was considered.

Results: The results show that 6 weeks combined training significantly decrease the plasma levels of chemerin (p=0.004), and SAA (p=0.009), but there was not any significant decrease on CRP levels(p=0. 476). So, it can be concluded that combined training will affect on some inflammatory markers in obese men and improve them.

Conclusion: One session combined training for 6 weeks significantly affects on plasma levels of chemerin and serum amyloid-A, however, it doesn't decrease the plasma levels of C-reactive protein (CRP).

Background: Influenza A viruses are globally important respiratory pathogens which cause a high degree of morbidity and mortality during annual epidemics. M2 protein which expressed on the viral surface facilitates virus entry to the host cells. The extracellular domain of M2 protein (M2e) consists of N-terminal 24 residue which shows remarkable conservation among all subtypes of influenza A viruses. In this study, we evaluated the immunogenicity of three tandem repeats of M2e along with different adjuvants in BALB/C mice model.

Materials and Methods: Recombinant protein (3M2e) was expressed in Escherichia coli and purified. Six weeks old BALB/c mice were immunized interdermally with three doses of 3M2e alone or supplemented with Alum/CpG motif as adjuvant. Control group was injected with PBS. Two weeks after the last immunization, specific anti-M2 was measured using ELISA method and finally mice were challenged with one lethal dose (LD90) of PR8 virus.

Results: The results showed that 3M2e can induce specific antibody alone. However, 3M2e protein supplemented with Alum-CpG induced higher level of specific antibodies, so that, there was a significant difference with 3M2e group (p<0.05). Anti-M2 antibodies mostly consisted of IgG2a subclass which considered as activity index of TH1 Cells. Moreover, this group showed enhanced protection against wild-type virus (survival rate=60%).

Conclusion: Applying Alum-CpG as a complex adjuvant may play a crucial role in integrating innate and acquisitive immunity. We increased density of M2e in combination with complex adjuvant and showed that this vaccine induced power immune responses and semi-protected mice against lethal challenge.

Background: Nef is one of the HIV-1 critical proteins, because it is essential for viral replication and AIDS disease progression and induction of immune response against it can partially inhibit viral infection. Moreover, a domain of the HIV-1 Trans-Activator of Transcription (Tat, 48-60 aa) could act as a cell penetrating peptide (CPP). In current study, cloning and expression of Tat-Nef fusion protein was performed in E. coli for the first time. The protein expression was confirmed by western blot analysis and was purified using reverse staining method.

Materials and Methods: In this experimental study, primarily, cloning of Tat-Nef fusion gene was done in pGEX6p2 expression vector. Then, the expression of Tat-Nef recombinat protein in E.coli BL21 (DE3) strain was performed by using IPTG inducer. The protein expression was confirmed by SDS-PAGE and western blotting using anti-Nef monoclonal antibody. Then, the recombinant fusion protein was purified from gel using reverse staining method.

Results: The results of PCR analysis and enzyme digestion showed a clear band of ~ 726 bp in agarose gel indicating the correct Tat-Nef fusion cloning in pGEX6p2 prokaryotic expression vector. In addition, a 54 kDa band of Tat-Nef on SDS-PAGE revealed Tat-Nef protein expression that western blot analysis using anti-Nef monoclonal antibody confirmed it.

Conclusion: The purified Tat-Nef recombinant fusion protein will be used as an antigen for protein vaccine design against HIV infection.

Background: Immunotoxin (IT) is a directed toxin containing two distinct sections (immune and toxin parts) covalently bond using specific chemical or peptide linkers. The aim of this study is to produce a recombinant and hybrid protein containing diphtheria toxin (DT) and granulocyte colony stimulating factor (G-CSF).

Materials and Methods: According to the structure of the first commercial recombinant immunotoxin (Ontak, hybrid protein containing DT fused interlukine2), gene encoding of DT and G-CSF was amplified using specific primers and plasmid template of pET-IDZ3 (pET21 harboring the gene encoding ontak immunotoxine) and pET-GCSF (pET23 harboring G-CSF), respectively. The DT-GCSF fusion protein produced using soeing PCR and specific primers. Finally, pET-DT-GCSF construction prepared using subcloning of DT-GCSF into pET21a(+) and confirmed by sequencing, SDS-PAGE and western blot technique.

Results: Gene encoding of DT-GCSF inserted into NdeI/EcoRI site of pET21 and the construction of strain producing DT-GCSF recombinant immunotoxin was confirmed using customary methods.

Conclusion: The cytokine fusion protein, DT-GCSF, could be used for the inhibition of G-CSF receptor bearing cancer cells.

Background: The renin-angiotensin system(RAS) has a major role in development of diabetic nephropathy and blocking of RAS by inhibitors and blocking of angiotesin receptors is standard treatment for preventing kidney disease and proteinuria. It is reported that VIT-D analogues are able to suppress renin exertion and improve proteinuria. The aim of this study is to evaluate the effect of VitD analogue (calitriol) on reducing proteinuria in patients with diabetic nephropathy.

Materials and Methods: In this clinical trial study, 132 eligible patients that had diabetic nephropathy and hadn’t vit D deficiency were selected. The patients were divided into two equal groups. First group received the combination of losartan 25mg twice daily and calcitriol 0.25mg and second group received losartan 25 mg twice daly alone for 3 months. The FBS, lipid profile, ESR-CRP BUN, Cr, HbA1c, Ca, P, and 24 hours urine protein were evaluated in all patients at beginning and end of study and the results were statistcally compared.

Results: The 24-hour urine protein in losartan and calitriol group was improved compared to losartan. This difference was statistically significant (p=0.003). As well as, in kidney function (BUN, Cr) in the losartan and calcitriol group compared to losartan alone was significantly improved(p<0.05).

Conclusion: Combination of calcitriol with angiotesin receptor blockers(ARBs) is more effective than ARBs alone in improvement of proteinuria and real function.

Abstract

Background: Carbon tetrachloride is one of the chemical toxins, disturbing bone marrow texture and changing the serum blood proteins. In this study, the protective effect of Avicennia marina leaf extract on bone marrow texture of rats induced by carbontetrachloride is investigated.

Materials and Methods: 42 male rats were divided randomly in to 6 groups: group induced by CCl₄ (carbon tetrachloride 1:1 with olive oil, 2 ml/kg single dose, i,p), sham(taking olive oil, 2ml/kg i,p single dose) and control (taking normal saline, 2ml/kg, i,p single dose ). Treated groups (1,2 and3): induced by carbon tetrachloride 1:1 with olive oil, 2ml/kg single dose and then after two hours treated by 200mg/Kg, 400mg/Kg and 800mg/kg AME /day for 96 hrs, i,p) After the examination the blood samples were collected from heart directly and WBC and blood proteins such as Albumin, total protein separation of serum and Sections sternum bone were analyzed. Data were analyzed by ANOVA and statistical significance differences were accepted at(p<0.05).

Results:  The necrotic bone marrow texture, WBC, serum Albumin and total protein of the treatment groups showed a significant increase rather than group induced by CCl₄(p<0.001).

Conclusion:  The Avicennia marina leaf has active antioxidant and flavonoids compounds which probably have protective effects on bone marrow texture from toxic agents such as CCl₄.

Abstract

Background: The aim of this study was to evaluate the effects of supplementation of Branched-Chain Amino Acids (BCAAs) plus carbohydrate (CHO) and whey protein plus CHO on muscle damage indices after eccentric resistant exercise.

Materials and Methods: Twenty four untrained healthy males participated in this study. They were randomly divided into three groups, BCAA +glucose (0.1+0.1g/kg) supplement group (n=8), Whey+glucose (0.1+0.1g/kg) supplement group (n=8), and placebo (malto dextrin 0.2g/kg) group (n=8). Each subject consumed a carbohydrate beverage with addition of whey protein or branched-chain amino acid or placebo 30 minutes before exercise in a randomized,double-blind fashion. Serum levels of Creatine Kinase (CK), Lactate dehydrogenase (LDH), and muscle pain were measured before, 24, 48, 72 h after exercise. Follow-up analyses included 1-way repeated measures ANOVAs, and Bonferroni post hoc comparisons.

Results: 24 h after test, serum levels of CK, LDH and muscle pain in both supplement groups were increased less than placebo group (0.015, 0.001 and 0.001, respectively). Also, the levels of CK and LDH showed significant changes in both intervention groups compared to placebo group at 24 h (0.001, 0. 015, respectively). Similarly, significant differences in the levels of CK and LDH between groups were observed.

Conclusion: These data indicate that muscle damage and pain after resistant exercise were reduced by an ingestion of either BCAA drink or whey protein drink.