Pathobiology Research
Year:2010 | Volume:13 | Issue:2|Papers Count:9

Objective: Helicobacter pylori is a widely distributed Gram negative bacterium that infects the human stomach and duodenum. Some antibiotic regimens are subjected to cure the infection but the cost of drugs, poor patient compliance and emerging of antibiotic-resistant strains are limiting the usefulness of these antibiotic therapies. Therefore, interest in developing a H. pylori vaccine is growing up rapidly. The aim of this study was to construct a recombinant vector containing fusion genes encoding a fragment of B subunit from Helicobacter pylori (H. pylori) urease (UreB332) and Helicobacter pylori adhesion A (HpaA) and expressed it in E. coli BL21, as well as determining its antigenicity as a vaccine candidate of H. pylori.Materials and Methods: The target genes encoding UreB332 and HpaA amplified from standard H. pylori chromosome by PCR, digested by restricted endonuclease enzyme and inserted into the prokaryotic expression vector pET28a (+) which was digested by corresponding restricted endonuclease enzyme. The target fusion protein was expressed in the BL21 (DE3) E. coli. Furthermore, UreB332-HpaA antigenicity was studied by western blotting after Ni-NTA agarose resin purification.Results: Enzyme digestion analysis, PCR and sequencing showed that the target genes were inserted correctly into the recombinant vector. The fusion protein UreB332-HpaA was recognized by the rabbit anti H. pylori polyclonal antibody and the human sera infected with H. pylori.Conclusion: Our results in addition to favorable properties of HpaA and UreB antigens, support the application of rUreB332-HpaA fusion protein, as a good candidate for the development of H. pylori vaccine.

Objective: This study was designed to investigate the antinociceptive effect of chronic administration of the flavonoid hesperetin in streptozotocin-diabetic rats using formalin and hot tail immersion tests.Materials and Methods: Rats were divided into 5 control groups, hesperetin-treated control, diabetic, sodium salicylate (SS) -treated diabetic, and hesperetin- and glibenclamide-treated diabetic groups. Hesperetin (10 mg/kg) was administered i.p. one other day 1 week after diabetes induction for 6 weeks. Finally, thermal pain tolerance and nociception were evaluated using hot water tail immersion and formalin tests respectively.Results: Diabetic rats exhibited a higher score of pain at both phases of the formalin test (P<0.05) and hesperetin-treated diabetic rats exhibited a lower nociceptive score at both phases of the test (P<0.05). Regarding thermal pain tolerance, diabetes significantly reduced tail immersion latency (P<0.01) and hesperetin treatment did produce a significant change in this respect (P<0.05).Conclusion: Chronic treatment with hesperetin for 6 weeks does mildly increase thermal pain tolerance and reduces chemical nociception in an experimental model of diabetes mellitus and this may be considered as an auxiliary treatment for diabetic hyperalgesia.

Objective: Hearing loss is the most frequent neurosensory defect in human. Mutations in GJB2 and GJB6 are responsible for 50% of autosomal recessive non-syndromic hearing loss (ARNSHL) cases. Here we report on the frequencies of GJB2 and GJB6 mutations and three large deletions spanning the GJB6 gene including Del (GJB6-D13S1830), Del (GJB6-D13S1854) and a>920 kb deletion in patients affected by ARNSHL referred to Kawsar's Human Genetics Research Center.Materials and Methods: In this study, 94 patients from 63 families with ARNSHL were investigated. Patient's homozygote for 35delG were screened and left out of the study and the remaining samples were analyzed by sequencing of GJB2 and GJB6 genes. Also the three large deletions spanning the GJB6 gene were analyzed by Real Time PCRResults: In this study we found GJB2 mutations in 13 families (20.6%) out of 63. The 35delG mutation was the most common mutation in the studied population (61.5%). Other GJB2 mutations were delE120, R127H, W24X, and V37I. The heterozygous or negative cases for the GJB2 mutations were screened for mutation in the GJB6 gene by sequencing and no mutation was observed. Also, we checked the three large deletions in GJB6, we found no mutations.Conclusion: Low frequency of mutations in the GJB2 gene implies that other genes may be involved in causing non-syndromic hearing loss in our country.

Objective: Polymerase chain reaction (PCR) is one of the most important progress in the field of molecular biology and diagnosis. Despite simplicity in concept, the reaction needs complex interaction between target sequence, primers, dNTPs and DNA polymerase for a successful amplification and diagnosis. For the detection of RNA viruses highly sensetive and specific technique is required. Hence amplification based on Nested PCR would improve sensitivity, and also use of one step reaction would decrease probable contaminations as reported previously in several studies. The aim of the current study was to develop a new, rapid and sensitive one step- one tube Nested PCR in a closed system, by using two novel coherent primers.Materials and Methods: In this study, a novel and special primer development method was used for one step- one tube reaction. After development and optimization, the assay was evaluated with known positive and negative controls.Results: The developed assay was performed on 50 HCV positive samples and 10 negative controls and 5 samples from each HIV, HBV, TTV (Torque Teno Virus) and GBV-C (Hepatitis G Virus: HGV). Based on the obtained results, sensitivity and specifity was calculated.48 out of 50 HCV positive samples showed expected band while none of the negative controls gave any band.Conclusion: Based on the specific primer design system which has been used in the current study; the inner primer was synthesized as complementary of routine PCR primers and was bound to the outer primers. Therefore, there is no probability for false priming in the both rounds of the reaction, hence there would be no nonspecific amplifications. Other advantages of this assay system were prevention of contamination which was due to one step- one tube reaction, decrease in duration and the cost of the reaction.

Objective: Considering the various evidences due to effects of trace elements on the outcome of pregnancy, we decided to assess the effect of these elements levels in maternal and cord serum on low birth weight.Materials and Methods: An Analytical cross-sectional study was performed on 344 pregnant women in labour and their neonates in Maryam, Akbarabadi and Vali-e-Asr hospitals in Tehran, Iran. Iron, zinc, copper, magnesium and calcium concentrations in cord and mother serum were measured. Collected data were analyzed by spss 13 software, using Chi- square and Logistic regression tests with significant level p<0.05.Results: The characteristics of studied subjects were: Mean age: 27.02±5.3 years, pregnancy duration: 38.9±1.3 weeks, number of low birth weight neonates: 13 (3.8%). Prevalence of trace elements deficiencies in mothers were: zinc: 26.7%, copper: 53.5%, magnesium: 72.4%, Iron: .6%, calcium: 49.4%, and in neonates were: zinc: 3.5%, copper: 41.6%, magnesium: 67.4%, Iron: 3.2% and calcium: 2.3%.In Chi- square test, a significant negative relationship between maternal calcium deficiency and low birth weight (p= .011) was found and logistic regression analysis showed a significant negative relationship between maternal calcium serum and low birth weight (odd: 0.27, %95CI: 0.09-0.77).Conclusion: Delivering low birth weight neonates were more common in mothers with calcium deficiency and other elements did not show any significant relationships with low birth weight. It is possible to find different results about the effect of these minerals on pregnancy outcome with more samples and in different settings. It is recommended to perform more research on effects of trace elements on neonatal and maternal pregnancy outcomes to help prevent mother and fetus mortality and morbidity.

Objective: The theraputic exercise is regarded as one of the most important theraputic interventions in the chronic low back pain patients. Stabilization exercise is among the most significant practical ones in these patients. The study of the effects of these exercises on the variability of trunk muscles postural strategies and the ability in normalizing these strategies are among the uncertainties that have remained unknown up to the present time. The purpose of this research is the study of the effect of stabilization exercises on the variability of postural control strategies in these patients.Materials and Methods: The study was performed on 21 (14 males, 7 females) recurrent non-specific low back pain patients. The electromyographic activity of Deltoid, Transverse Abdominis/Internal Oblique, External Oblique and Erector Spinae muscles of each person was recorded in 75 rapid arm flexion with maximum acceleration. Then, 14 individuals were placed in the experimental group and 7 in the control group randomly. The experimental group performed isolated abdominal hollowing and bracing exercises using pressure biofeedback with the supervision of a physiotherapist in different positions including supine, sitting on a chair, standing still and standing with rapid arm raises with maximum acceleration once a day, for 20 sessions. The variables studied in this research were evaluated in both experimental and control groups after the training sessions. The standard deviation of the trunk muscles onset latencies relative to deltoid muscle was statistically analyzed by a dependent t-test in both groups.Results: The results revealed that performing the stabilization exercises increases the variability of the timing of anticipatory postural adjustments in TrA/IO in the experimental group (P= .037), while there were no significant changes in the variability of EO (P= .0346) and ES (P= .673) muscles in these patients. The variability of the timing of the anticipatory postural adjustment in TrA/IO (P= .199), EO (P= .702) and ES (P= .465) muscles did not show any significant changes in the control group.Conclusion: The study of the results showed that the reduction in the variability of postural control system in recurrent non-specific low back pain patients as a factor that causes the continuity of back pain can be improved by goal-directed stabilization exercises.

Objective: Saffron is the dried stigmas of Crocus sativus L. which has various therapeutic properties in addition to its use as a spice. More than 80% of the world present production of saffron which is about 190 tons is produced in Iran. To compare the quality of saffron' from different parts of the world, researcher used one sample from each country. Since cultivation of saffron is done in different geographical areas of Iran, it seems that the results of these studies, using only one sample of Iranian saffron, are not consistent with the scientific and statistical rules. More over the results of therapeutical studies using saffron extracts from different sources with different concentrations of effective elements would not give reproducible results. As an affirmative reason, here we compared the saffron’s important components, crocin, picrocrocin and safranal, from different packages produced by some companies.Materials and Methods: Five certified saffron packages, prepared by different companies named: Ehteshamiyeh, Tarvand, Abbaszadeh, Sabagh and Novin-Saffron were analyzed using HPLC with an UV/Vis detector. To avoid the conflict of inerest, these samples were randomly labeled as 1 to 5.Results: Results indicated that all of the samples contained these ingredients at the standard levels, but the highest level of crocin as a carotenoid was seen in the sample 3, and sample 1 and 2 contained the highest concentrations of monoterpene aldehydes, picrocrocin and safranal, respectively.Conclusion: According to the obtained results and the vast area under the saffron cultivation with various climates, it is confirmed that one sample of saffron is not a measure of all of the Iranian saffron. Our research is continuing.

Objective: Cryptosporidiosis is a parasitic disease that is causing small protozoan genus Cryptosporidum and transmission take place through fecal- oral by direct contact or indirectly through food or drink. The aim of this study was detection of anti-Cryptosporidum parvum Immunoglobulin IgG, in newborn BALB/c infected with C. parvum.Materials and Methods: Oocysts of C. parvum were obtained from the feces of diarrheic lambs and following purification they were suspended in 2.5% aqueous potassium dichromate solution and stored at 4oC. Forty suckling BALB/c (3-4 days old) were divided in 8 groups (4 case groups and 4 control groups) each group consist of 5 suckling BALB/c. The mice in case groups were infected oraly with 105 C. parvum oocysts, and the mice in control groups served as non-infected. Blood samples were collected at 6, 9, 12 and 16 days post-infection (pi). Immunoglobulin IgG were extracted by salting out method and confirmed with SDS-PAGE.Results: Antibodies were analyzed by western blot and increased secretion of IgG was confirmed in neonatal mice infected with Cryptosporidum oocysts. Mean OD of Immunoglobulin IgG increased from 0.350±0.099 to 0.6776±0.099 in case groups but in control groups the increase was from 0.244±0.016 to 0.322±0.16 (P<0.05).Conclusion: The type of antibody in neonetal mice infected with Cryptosporidum oocysts was IgG which is secreted against external memberane of oocysts. Significant differences in neonatal mice case groups as compared with the control groups were observed.

Objective: Several vaccines against HIV have been investigated but none has been approved as an effective HIV vaccine. An approach that could induce stronger immune response against the pathogen is utilizing a multi-epitopic vaccine. This strategy was used in the design of several vaccines and resulted in improved immune responses.Materials and Methods: In this study a multi-epitopic fusion peptide including parts of HIV-1 Nef and P24 as a vaccine candidate was injected into mice and immune humoral responses measured with total antibody and IgG sub-classes using ELISA. Also measurement of cellular immune responses through evaluation of spleen cells proliferation response using MTT and cytotoxicity by LDH were performed. Finally, the cytokine pattern of IFN-g and IL-4 were also determined with ELISA.Results: The results indicate that candidate vaccine stimulated mouse splenic lymphocyte proliferation response and also induced strong cytotoxicity responses. Analysis of humoral immune response has shown that the candidate vaccine has induced specific antibody production mainly of the IgG2a sub-class. Also cytokine pattern evaluation has shown that IFN-g secretion was dominant.Conclusion: The use of immunogen and conserved epitopes from P24 and Nef induced strong humoral and cellular immune responses and this construct could be candidate for further studies in animal models.