Iranian Journal of Basic Medical Sciences
Volume:17 Issue: 8, Aug 2014

Helicobacter pylori infection occurs worldwide, but the prevalence of this infection varies greatly among different countries and population groups. The aim of this study was to determine the seroprevalence of anti-Helicobacter pylori and anti-cytotoxin-associated gene A (CagA) antibodies in asymptomatic healthy population in the center of Iran and to investigate the relation with different parameters. Totally, 525 individuals aged 17-60 years were enrolled in study. The serum samples of participants were tested for anti-H. pylori IgG and anti-CagA IgG by enzyme-linked immunosorbent method (ELISA). ABO blood grouping was also done by hemagglutination test. The seroprevalence of anti-H. pylori IgG was 74.2% and their rates increased with age. The seroprevalence of anti-H. pylori IgG was higher in males (74.6%) than in females (71.6%). There was statistically inverse association between H. pylori infection and education level (P=0.04) and marital status (P=0.000). The most prevalent blood group was type AB with positive Rh-phenotype (82.4%). In H. pylori infected individuals the seroprevalence of anti-CagA antibody was 46.9%. The seroprevalence of anti-CagA IgG was in males 48.6% and in females 31.6%. There was no statistically significant association between anti-CagA IgG positivity and age, occupation, socioeconomic status, ABO blood groups and Rh status. These resultsshowed that H. pylori infection was common in the asymptomatic individuals. Almost half of the infected individuals acquire CagA-positive strains of H. pylori. Moreover, it seems that males are more susceptible to infection with CagA-positive strains compared to females.

The aim of this study was to improve flowability and compressibility characteristics of starch to use as a suitable excipient in direct compression tabletting. Quasi-emulsion solvent diffusion was used as a crystal modification method. Corn starch was dissolved in hydrochloric acid at 80˚C and then ethanol as a non-solvent was added with lowering temperature until the formation of a precipitate of modified starch. Flow parameters, particle size and thermal behavior of the treated powders were compared with the native starch. Finally, the 1:1 mixture of naproxen and each excipient was tabletted, and hardness and friability of different tablets were evaluated. Larger and well shaped agglomerates were formed which showed different thermal behavior. Treated starch exhibited suitable flow properties and tablets made by the treated powder had relatively high hardness. It was found that recrystallization of corn starch by quasi emulsion solvent diffusion method could improve its flowability and compressibility characteristics.

Typhoid fever is a dreadful disease of a major threat to public health in developing countries. Vaccination with bacterial immunodominant components such as surface proteins may prove as a potent alternative to live attenuated vaccines. InvH, an important part of needle complex in type three secretion system (TTSS) plays important role in efficient bacterial adherence and entry into epithelial cells. In this work we used a 15 kDa recombinant InvH protein of Salmonella enteric serovar Enteritidis to provoke antibody production in mouse. The mice were immunized by recombinant InvH and challenged with Salmonella typhi. Histopathology of spleen and liver were studied. The immunized mice showed a significant rise of antibody after the second booster. The immunization induced protection against high doses of S. typhi. The bacterial challenge with sera showed significant protection against challenge dose of 2×109 CFU. Immunized sera reacted with S. typhi markedly. Immunoreaction of bacterially infected sera and InvH protein was significantly higher than the control group. Bacterial loads of S. typhi in spleen was more than liver. Decreased bacterial load was evident in immunized mice after 7 days. Histological examination of the liver showed the immunized mice liver remained unaffected. Efficacy of the virulence protein, InvH, in inhibition of this phenomenon by active immunization was shown here. It may be concluded that InvH, as an antigen, can develop protection against S. typhi infections. InvH may be exploited in protective measures as well as a diagnostic tool in Salmonella infections.

To culture thein vitro mouse embryonic stem cells (mESCs) and to direct their differentiation to germ-line cells; in present study we used a vector backbone containing the fusion construct Stra8-EGFP to select differentiated ES cells that entered meiosis. Retinoic acid was used to differentiate embryonic stem cells to germ cells. A fragment of Stra8 gene promoter (-1400 to +7) was inserted in ScaI/HindIII multiple cloning site of pEGFP-1 vector. Theelectroporationwas done on embryonic stem cells and positive colonies were selected as puromycin-resistant after three weeks of treatment with puromycin. All-trans retinoic acid (RA) was used for differentiation of mESCs at final concentration of 10-5M. The expression of protamine 1 (Prm1) gene was checked as post meiotic marker in differentiated mESCs after 5, 10, 15, 21 and 30 days after RA induction. The PCR amplification by specific primers for Stra8-EGFP fusion gene was detected in DNA sample from mESCs after electroporation and puromycin treatment. GFP-positive mESC colonies were observed after 72 hr RA induction. The protamine 1 gene was expressed after 21 days of RA induction. In this study, we demonstrated the in vitro generation of mouse embryonic stem cells to germ cells by using a backbone vector containing the fusion gene Stra8-EGFP. The Stra8 gene is a retinoic acid-responsive protein and is able to regulate meiotic initiation.

Osteoarthritis (OA) is globally one of the most common diseases from the middle age onwards. Cartilage is an avascular tissue therefore it cannot be repaired in the body. Conservative treatments have failed as a good remedy and cell therapy as a decisive cure is needed. One of the best and easily accessible cell sources for this purpose is adipose-derived stem cells which can be differentiated into chondrocytes by tissue engineering techniques. Chemical and physical inducers have a key role in stem cell – chondrocyte differentiation. We have tried to determine the role of electric fields (EF) in promoting this kind of chondrogenesis process. Human adipose derived stem cells (ADSCs) were extracted from subcutaneous abdominal adipose tissue during cesarean section. A high frequency (60 KHz) EF (20 mv/cm), as a physical inducer for chondrogenesis in a 3D micromass culture system of ADSCs was utilized. Also, MTT, ELISA, flow cytometry, and real-time PCR techniques were used for this study. We found that using physical electric fields leads to chondrogenesis. Furthermore, results show that using both physical (EF) and chemical (TGFβ3) inducers simultaneously, has best outcomes in chondrogenesis, and expression of SOX9 andtype II collagen genes. It also causes significant decreased expression of type I and X collagen genes in pure EF group compared with control group. The EF was found as a proper effective inducer in chondrogenic differentiation of human ADSCs micromass culture.

Obesity considered being a low-grade inflammatory disease. The objective of this study was to examine the association between inflammatory markers (IM) including C-reactive protein (hs-CRP), Interleukin-6 (IL-6), and homocystein (Hcy) and obesity[F1] -related factors (e.g. BMI, waist, hip) in adult participants of Tehran lipid and glucose study (TLGS). In this cross-sectional study, 352 individuals (132 men and 220 women), age ≥19 years, were randomly recruited from participants of TLGS population[F2]. The serum levels of hs-CRP, IL-6, Hcy were determined using the enzyme linked immunosorbent assay (ELISA) method[F3]. Variables were compared by sample t-test. Bivariate linear correlation was estimated using Pearson’s correlation coefficient. Linear regression analysis was applied to investigate the association between IMs and anthropometric and biochemical variables. The mean age of participants was 46.1±16.1 years. abdominal obesity was present in 199(56.5%) individuals. levels of hs-CRP and IL-6 increased in the abdominally obese group (1507±3.3 vs. 577.8±4.3 ng/ml P<0.001) (3.6±3.3 vs. 1.9±3.8 pg/ml P< 0.001), and in the same group, the best predictors for hs-CRP, IL-6 and Hcy were waist (WC), waist to height ratio (WHtR) and wrist respectively; hip and WHtR were the best predictors for Hcy and hs-CRP in the normal group. A linear augmentation in hs-CRP and IL-6 levels was observed in association with obesity categorizes. This study provides evidence that abdominally obese individuals had higher levels of IMs. Wrist, waist and WHtR were the best predictors for Hcy, hs-CRP and IL-6 respectively in this group.

Short tandem repeat (STR) loci are the most informative DNA genetic markers for attempting to individualize biological material for application in paternity and forensic cases. Blood samples were collected and the total genomic DNA was extracted. The DNA samples were used for genotyping VWA and TPOX STR loci using PCR and polyacrylamide gel electrophoresis. This report presents allele frequency data and parameters of biological or legal interest, such as heterozygosity value, polymorphic information content (PIC), genetic diversity index (GD) and population parameter (θ) in Arab and non-Arab population of Khuzestan province (Iran) for the loci VWA and TPOX. Blood samples (N= 392 for VWA and N=308 for TPOX) were collected from individuals unrelated throughout Khuzestan province. The loci were genotyped using the polymerase chain reaction (PCR) followed by polyacrylamide gel electrophoresis (PAGE) and silver staining. Chi-square test showed that neither STR loci were in agreement with the Hardy-Weinberg equilibrium. The examined STR loci in this study have proven a relatively high genetic variation in the Iranian population. The data could be used for construction of a forensic genetic database for the Iranian population.

Leptin receptor (LEPR) is a member of the class I cytokine receptor super-family that is known implicated in the initiation and progression of breast cancer. We have investigated the effect of Q223R polymorphism on the breast cancer susceptibly in a sample of Iranian subjects. We utilized a polymerase chain reaction (PCR) restriction fragment length polymorphism (RFLP) method to investigate the association ofLEPRQ223R polymorphism with breast cancer risk in a case control study consisting of 100 breast cancer cases and 100 controls without breast cancer. Serum levels of leptin and soluble leptin receptor (sOB-R) were measured by ELISA method. The genotype (QQ, QR, and RR) distributions were 25, 56, and 19 % in breast cancer cases and 54, 40, and 6% in controls, respectively. The frequency of 223 RR genotype was significantly elevated in breast cancer cases as compared to controls (χ2= 20.072, P<0.001). Similar significance differences were also found in allele frequencies for Q and R between two groups (χ2= 19.027, P< 0.001). Additionally, there weresignificant association between Q223R genotypes and breast cancer risk; homozygotes for RR genotype (OR= 6.840; 95% confidence interval [CI] = 2.434-19.218), heterozygotes for QR (OR=3.024; 95% CI = 1.620-5.644, P = 0.001), and QR+RR genotype (OR= 3.522; 95% CI = 1.934-6.414, P < 0.001), respectively. Our results showed thattheLEPRQ223R polymorphism is associated with increased breast cancer risk as well as tumor grade in a sample of Iranian subjects.

Macrolide resistant Streptococcus pneumoniae pose an emerging problem globally. The aim of this study was to investigate the prevalence of ermB and mefA genes (macrolide resistant genes) by polymerase chain reaction (PCR) method and to detect drug resistance patterns of S. pneumoniae isolated from clinical samples to macrolides and other antibiotic agents by E-test method. Fifty five isolates of S. pneumoniae were obtained from clinical samples with microbial tests. The antibiotic susceptibility of isolates for erythromycin, azithromycin, clarithromycin, ceftazidime, ciprofloxacin and vancomycin were determined by E-test method. Genotypic antibiotic resistance pattern was determined by PCR with primer designed for ermB and mefA genes. The number of S. pneumoniae isolates resistance to erythromycin, azithromycin, clarithromycin, ceftazidim, ciprofloxacin were 25.5%, 18.2%, 16.4%, 21.8% and 10.9%, respectively while no resistance to vancomycin was observed. The macrolide resistance genes of ermB and mefA were found in 10.9% and 18.2% of the isolates, respectively. The result of the current study suggests the necessity of evaluation the changes in MIC[FA1] [a2] (minimum inhibitory concentration) values as well as genetic mutations to estimate the prevalence of the resistance antimicrobial agents in S. pneumoniae.

The effect of litter size and suckling intensity on the expression of KiSS-1 mRNA in the arcuate nucleus (ARC) of rats were evaluated. Thirty two pregnant and four non-lactating ovariectomized (as control group) rats were used in this experiment. Lactating rats were allotted to eight equal groups. In three groups, litter size was adjusted to 5, 10, or 15 pups upon parturition and allowed to suckle their pups continuously by 8 days postpartum. In the other three groups, litter size was adjusted to five upon birth; the pups were separated from the dams for 6 hr on day 8 postpartum, after which the pups were allowed to suckle their dams for 2.5, 5, or 7.5 min prior to killing the dams. Two groups of lactating rats with either 10 or 15 pups were separated from their pups for 6 hr on day 8 postpartum, after which the pups were allowed to suckle their dams for 5 min before the dams were killed on day 8 postpartum. The ARC was removed and the expression of KiSS-1 mRNA was evaluated, using real-time PCR. The expression of KiSS-1 mRNA in the ARC was decreased as the litter size and intensity of suckling stimulus were increased. The effect of suckling intensity on the expression of KiSS-1 mRNA was more pronounced than that of litter size. Increased litter size and suckling intensity decreased KiSS-1 mRNA expression in the ARC which may contribute to lactation anestrus in rat.

Buthionine sulfoximine (BSO) inhibits synthesis of glutathione as the main intracellular antioxidant. The aim of the present study is to investigate the effect of BSO-induced oxidative stress on histological structure of testis, testosterone secretion and semen parameters. Thirty male BALB/c mice were divided into 3 groups. In control group, the mice did not receive any chemical. In the experimental group, the mice received 2 mmol/kg BSO for 35 days. In the sham group, the mice received the solvent of BSO (0.9% saline). After the treatment, the mice were sacrificed. Their testes were fixed in Buein’s fixative, embedded in paraffin and prepared for histological studies. To assess semen parameters, the sperms were collected from cauda epididymis. Blood samples were used for determination of super oxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GPX), glutathione (GSH), catalase (CAT) and the serum testosterone level. The data analyzed using ANOVA and Dunnett''s tests and SPSS software, version11.5. P- values at 0.05 level considered significant. Data showed that in experimental group in comparison to control group; the concentration of CAT, GPX, SOD,GSH and the total level of testosterone is reduced while MDA level is increased significantly. The number of sperms with progressive motility were decreased (P<0.001) but sperms with abnormal morphology were increased (P<0.001). Histological studies revealed that the values for tubal differentiation index and spermatogenic index in experimental group were reduced (P<0.001). It is concluded that exposure to oxidative stress induced by BSO could affect testicular structure and semen parameters.

Hyperglycemia, oxidative stress and apoptosis have key roles in pathogenesis of diabetic neuropathy. There are local renin-angiotensin systems (RASs) in different tissues such as neural tissue. Local RASs are involved in physiological and pathophysiological processes such as inflammation, proliferation and apoptosis. This study aimed to investigate the role of local renin-angiotensin system on high glucose-induced cell toxicity, apoptosis and reactive oxygen species (ROS) production in PC12 cells, as a cell model of diabetic neuropathy. PC12 cells were exposed to a high glucose concentration (27 mg/ml), captopril (ACE inhibitor), telmisartan and losartan (AT1 antagonists), and also PD123319 (AT2 antagonist) were administered before and after induction of high glucose toxicity. Then cell viability was assessed by MTT assay and apoptotic cells and intracellular ROS production were detected by annexin V-propidium iodide and DCFDA, respectively, using flow cytometry. High glucose concentration decreased cell viability, and increased apoptotic cells. Intracellular ROS production was also increased. In PC12 cells pretreatment and treatment by the drugs showed a significant improvement in cell viability and reduced apoptosis in captopril, telmisartan and PD123319 but only captopril and telmisartan were able to reduce ROS production. Losrtan significantly lowered ROS but didn’t show any improvements in cell viability and apoptotic cells. The results of the present study showed that RAS inhibitors reduced cell toxicity and apoptosis and ROS production was induced by high glucose. It may be suggested that local RAS has a role in high glucose toxicity.

To know whether Scurrula atropurpurea is able to modulate total plasma nitrate/nitrite levels, decrease endothelial damage, and increase endothelial progenitor cells (EPCs) in hypertensive rats. The rats were divided in 5 groups: control (normotensive) group, Desoxy cortico sterone (DOCA)-salt hypertensive group, and three DOCA-salt hypertensive groups. All 5 groups received methanolic extract of S. atropurpurea (MESA) at a dosage of 50; 100; and 200 mg/KgBW. Serum nitric oxide (NO) was assayed by colorimetric. Circulating endothelial cells (CECs) and EPCs were assayed using flow cytometry. The administration of MESA100 and MESA200 elevated the total plasma nitrate/nitrite levels but cannot reach the level in control group. MESA100 and MESA200 also elevated the EPCs number compared with hypertensive group. The administration of MESA significantly (P< 0.05) decreased the CECs number compared to hypertensive groups. Methanolic extract of S. atropurpurea is able to modulate total plasma nitrate/nitrite levels and diminish endothelial damage via increasing EPCs.

Sea cucumber derived bioactive compound is considered efficient in treatment of bone disorders. This study was performed to evaluate the effect of this extract on differentiation of rat bone marrow mesenchymal stem cells (rBMMSc) into osteogenic lineage. Isolated rBMMSc were grown in DMEM supplemented with 10% FBS. The cells were exposed to different concentration of extract. After 21 days, Alizarin red staining, alkaline phosphatase assay and RT-PCR were performed. The results were analyzed by ANOVA software and P value <0.05 was considered significant. Morphological methods revealed that appropriate concentrations of extract increased osteogenic differentiation in a dose-dependent manner. RT-PCR revealed that extract without or with osteogenic medium due to osteopontin expression had a potential role in osteogenesis. Based on our data it concluded that S. cucumber extract stimulated Bone marrow mesenchymal cells to differentiate into osteogenic lineage without existence of osteogenic medium