مجله فیض
سال سیزدهم شماره 3 (پیاپی 51، پاییز 1388)

Considering the immunosuppressive effects and prevalent mutations in some HCV antigens, induction of CD8+ T cell responses is focused on conserved and critical epitopes which as a multi-epitope vaccine can prevent the chronic nature of the disease. Two immunodominant HLA-A2-restricted human epitopes (E2614-622 and NS31406-1415) and two H-2d-restricted mouse epitopes (core132-142 and E2405-414) were designed in a sequential tandem, predicted by immunoinformatic analyses. Following the synthesis, related nucleotide sequence was cloned into the pcDNA3.1 vector with and without the fusion of hepatitis B surface antigen (HBsAg). Two constructed plasmids (pcDNA3.1.HPOL and pcDNA3.1.POL, respectively) were evaluated for the protein expression and secretion in Cos-7 cell line. After the vaccination of BALB/c mice (n=6 in each group) with different DNA and peptide immunization regimens, CD8+ T cell activity as well as the type and protective potency of the induced responses were evaluated. Despite the induction of epitope-specific responses in pcDNA3.1.POL injected mice, the group immunized with pcDNA3.1.HPOL indicated a significant increase in the number and activity of CD8+ T cells (P<0.05). Peptide boosting of this group (formulated in two human-compatible adjuvant) still led to the more activation of CD8+ cells, induction of Th1 response and the inhibition of tumor model growth (P<0.05). Fusion of HBsAg as a particle-forming sequence and the source of helper epitopes along the DNA-prime/peptide-boosting immunization regimen are proposed as two promising strategies to improve the CTL multi-epitope vaccines against HCV.

Brucellosis is one of the five common bacterial zoonoses caused by a gram negative, non-spore forming, and facultative intracellular bacterial organism belonging to the genus Brucella. Although brucellosis is considered as a health problem for both men and domestic animals in many countries, any licensed human vaccine has not been designed and produced for it yet. To overcome the problem, currently, antigenic determinants of Brucella cell wall e.g. outer membrane proteins [OMPs] and lipopolysaccharide [LPS] are considered as potential candidates to develop subunit vaccines. Brucella abortus S99 used in the present study is obtained from the standard bacterial collection of Institute Pasteur of Iran. OMPs were extracted by deoxycholate extraction technique and further purification performed by sequential centrifugation and ultracentrifugation. Protein concentration was determined using the Nanodrop ND–10000 spectrophotometery. SDS-polyacrylamied gel electrophoresis (SDS–PAGE) was performed to determine the electerophoretic pattern and the molecular weight of the extracted OMP samples. OMPs concentration of B.abortus S99 has been measured and reported as 6.27 mg/ml. SDS-PAGE analysis indicated one protein band in the range of 36-38 kDa which would be classified as the porins of B.abortus S99. Extraction of B.abortus S99 OMPs with the applied method in the present study produced a satisfactory yield of OMPs. These proteins belonging to the second group of OMPs, called porins.

Azathioprine (AZA) is the most widely used immunosuppressive drug for preventing graft rejection and autoimmune disease. However, the therapeutic treatment induces several side effects such as toxicity to bone marrow, pancrease, liver and gastrointestinal tract. One of the major functions of Vitamin A is to act as a natural antioxidant by scavenging free radicals. Considering the kind of Azathioprine-induced damage in Liver tissue, we decided to study the protective effect of Vitamin A against Azathiprine-induced toxicity. Forty Male Wistar rats were divided into 4 groups (each group contains 10 rats). Group 1 was control group and only took normal saline. Groups 2 & 3 were administrated daily use of Vitamin A for 7 days I.M. and Group 4 was administrated with normal saline instead of Vitamin A in same condition as groups 2&3. In the last day groups 3 & 4 were administrated with single dose of AZA, 15 mg/kg (IP). After 24 hours, we took the animals blood and tissue samples and studied them for biochemical and pathological examinations. This study showed that Azathioprine-induced damage on liver in group 3 is less than that in group 4 while the function of organ in group 3 is nearly the same as control group. Also vitamin A decreases Azathioprine-induced hepatotoxicity in rats. Regarding importance of Azathioprine-induced damage, the usage rate of this drug in medicine, and the results of this study, we suggest that co-administration of Azathioprine and vitamin E decreases the toxicity of this drug.

High lipid serum level was a risk factor for cardiovascular disease. This study was performed to evaluate the effects of Lovastatin and exercise on hyperlipidemic patients at Kashan Shahid Beheshti Hospital during 2005–6. This randomized clinical trial study was carried out on hyperlipidemic patients (age>30 years). After registering the personal data of the patients the participants were selected based on the inclusion criteria (HDL<35 mg/dl, LDL≥160 mg/dl, TG≥200 mg/dl, Chol≥200 mg/dl). the Lovastatin group (40 mg/daily) and the exercise group (waking 35 minutes/day, 3 times a week). The clinical tests were repeated for six consecutive weeks. Data were analyzed using t-Test, Wilcoxon Signed Rample Test, Kolomogorou-Smirnov Test, Mann-Whitney Test, levens Test. from Ninety-six out of 150 cases were hyperlipidmic signifying that only 80 (59 male, mean age 55.12±8.4 case and control 55.35±9.2 years) out of 150 hyperlipidemic patients were qualified for the study. The mean decrease in TG level between the Lovastatin and exercise groups were 21.4 (P=0.2) and 20.5 mg/dl (P=0.2), respectively. The mean decrease in cholesterol level between Lovastatin and exercise groups were 41.3 and 1.5 mg/dL (P=0.001). The mean decrease in LDL level between Lovastatin and exercise groups were 35.8 and 8.5 mg/dl (P<0.001), respectively. Finally, the mean increase in HDL level between Lovastatin and exercise groups were 5.4 and 0.4 mg/dl. exercise may be effective for decreasing the LDL. While Lovastatin could decrease the total cholesterol TG and increase HDL levels, using a combination is recommended for hyperlipidemic patients.

The differentiation of pleural effusions as exudates or transudates is the first step in the diagnosis of pleural effusions. The aim of this study was to evaluate the usefulness of pleural adenosine deaminase (PADA), for separating transudative pleural effusion from exudative pleural effusion. Using light's criteria, after careful evaluation of all relevant clinical and biochemical parameters of pleural fluid and serum, classically effusions were classified to transudative or exudative. The means and standard deviations of PADA, and pleural/serum ADA (P/S ADA) ratio were calculated for transudative and exudative effusions. The best cut-off values for each test were identified by using the receiver operating characteristic (ROC) curve. We compared PADA levels between transudative and exudative patients with pleural effusion. According to the criteria used, 32 patients were included in the transudate group and 73 patients in the exudate group. Pleural fluid ADA levels were significantly lower in the transudate group than in exudate one (16.21±6.10 U/lit vs. 22.24±7.50 U/lit P<0.001). Also the mean pleural fluid/serum ADA ratio were (P<0.0001) lower in transudates as compared to exudates (0.76±0.32 vs. 0.91±0.26 P=0.021). Using a cut-off point of 14.5 IU/L, the sensitivity and specificity of pleural ADA in the diagnosis of exudates was computed to be 84.7% and 48.3% respectively. At a cut-off point 0.75, pleural fluid/serum ADA ratio was found to have sensitivity and specificity of 73.2% and 65.5%, respectively. Measuring PADA is suggested as a useful biochemical marker in differentiating between pleural exudates and transudates.

Considering the significance of early diagnosis of cervical cancer and low sensitivity of conventional pap smear tests, this study was carried out to evaluate the adequacy of papanicolaou smear taken by liquid-based thin layer method compared to the conventional methods for women referring to obs/gynecological clinics of Kashan (March to October 2008). This cross-sectional descriptive study was done on pap smear specimens (n=400) of women referring to gynecological clinics. Randomly for half of samples, pap smear was prepared by manual liquid-based thin layer (direct to vial) method and the conventional method for half of the samples. Samples were prepared using cytobrush and interpreted based on Bethesda (2001) criteria. In each of sampling methods both satisfactory and unsatisfactory cases were identified and analyzed using Chi-square test. In both methods, the number of satisfactory and unsatisfactory smears were 191 (%95.5) and 9 (%4/5) respectively, signifying no difference between methods. The factors for unsatisfactory smear in conventional smear were exudates (8 cases [(88.9%]) blood (1 case [11.1%]) and in liquid based samples the lower cellularity (9 cases [100%]). The number of satisfactory cases in liquid-based thin layer method was the same as the conventional method.

Although type-2 mellitus diabetes is the most common type of diabetes, it's main cause yet to be identified. Chemokines and their receptors are probable effective systems on diabetes. CCR5 is a chemokine receptor playing an important role in immune responses. Studies showed that the known 32 mutation in CCR5 gene leads to disorder in the expression and function of this receptor. Hence, this project aimed to analyze the known 32 mutation in CCR5 chemokine receptor. Blood samples were collected from 200 type 2 diabetic patients and 300 healthy adult controls on EDTA pre-coated tubes. DNA was extracted using commercial kit. DNA samples were analyzed for 32 mutation by Gap-PCR in diabetic patients in compared to controls. The demographic information were collected through questionnaire. Our results showed that none of the diabetic patients displayed CCR5 32 mutation. While 2 out of 300 healthy controls had heterozigotic form of this mutation. Statistical analysis didn’t show any significant difference between the two groups. Several different studies analyzed the relation of this mutation with different types of diseases including diabetes. All studies failed to find a relation between this mutation and type 2 diabetes. Since these studies were performed in different geographical points and races, we studied this mutation in Rafsanjanese population. Based on the results of our study it could be probably concluded that this mutation does not play a key role in the establishment of type 2 diabetes.