KOWSAR MEDICAL JOURNAL
Year:2007 | Volume:12 | Issue:2|Papers Count:12

Objective: Study of the chondrogenesis of the marrow derived murine mesenchymal stem cells (MSCs) embedded three-dimensionally within an alginate gel. Materials and Methods: 4-6 week old NMRI mice were killed, the bone marrow cells were harvested, counted and 500 cells of which were plated per each well in 6-well-cell-culture plate. Fibroblastic cells were then purified and expanded through two successive subcultures. In order to prepare three-dimensional culture systems, 10 million of passaged-2-fibroblastic cells were suspended in 10 ml of alginate solution and the cell suspension were divided into 10 equal part each of which were placed per well of 12-well-culture plate. MSCs-alginate suspension were exposed to CaCl solution by which the gelation process was occurred. After that the CaCl was removed and the cultures were provided by a chondrogenic differentiation medium containing 10ng TGF-B3 and 10 ng BMP-6 for 3 weeks after which the occurrence of the differentiation were evaluated by toloidin blue staining, RT-PCR analysis and transmission electron microscopy. In present study the mesenchymal nature of the isolated cells were evaluated by differentiation into bone and adipocytic cell lineage. Results: In primary culture, variety of cell morphology ranging from the polygonal cells to the elongated, flattened and spindly–shaped cells was observed. The pure fibroblastic cells were not appeared until time the first subculture was performed. Enough number of the fibroblastic cells was then available for conducting the remaining part of experiment after the second round of cell passage was done. The result of the differentiation evaluation by toloidine blue staining method indicated that the MSCs in alginate assumed a rounded morphology with a lacunae around them. The cells have been surrounded by a methachromatic matrix. RT-PCR analysis revealed that the mRNA of collagen II, X and agreacan have been largely produced in differentiated cells. Ultra thin sections showed the cells with well-developed secretive organelles indicative of their active chondroblastic state rather than non-active chondrocytic state. Conclusion: MSCs could acquire cartilage-cell phenotype when being cultivated within the alginate gel in a three-dimensional culture system in the absence of cell condensation, an important prerequisite for chondrogenesis during development.

Objective: In the present study, the effects of intra-ventral tegmental area administration of L-arginine, a nitric oxide precursor, and NG-nitro-L-arginine methyl-ester (L NAME), a nitric oxide synthase inhibitor, on the acquisition and expression of morphine-induced place conditioning in morphine-sensitized rats were studied. Materials and Methods: Subcutaneous (s.c.) administration of morphine (5 mg/kg) induced conditioned place preference. Repeated pretreatment of morphine (5 mg/kg, s.c.) followed by 5 days no drug treatment, increased conditioning response induced by morphine (0.5 mg/kg, s.c.).Results: Intra-ventral tegmental area administration of L-arginine (0.01, 0.1 and 0.3 mg/rat) has no effect on the acquisition and/or expression of morphine place conditioning in non-sensitized and sensitized rats. Intra-ventral tegmental area injections of L-NAME (0.1 mg/rat) reduced the expression but not the acquisition of morphine place conditioning in the sensitized animals. Conclusion: The results indicate that nitric oxide (NO) within the ventral tegmental area is not involved in the acquisition and expression of morphine place conditioning in morphine-sensitized rats.

Objective: Since Candida albicans is the main cause of infection within candida factors, and ‎regarding that Candida glabrata replacement as a producing factor is the second and third within ‎Candida factors, the aim of this research was to evaluate the protein pattern of cell wall of ‎Candida albicans and Candida glabrata. One of the origins which can affect in causing disease is ‎organism protein especially cell wall protein. The objective of this research is firstly to acquire ‎Protein pattern of cell wall in approved factors of Candida albicans and Candida glabrata in a ‎seprate manner, and secondly to obtain protein pattern of cell wall of Candida albicans gathering ‎from clinical resources. Resulting information can present of similarities and differences ‎between obtained patterns and standard patterns. The results of total protein patterns of cell wall ‎can be used in comparing them with patterns of cytoplasmic proteins and the results can be used ‎at In vitro and molecular studies.‎ Materials and Methods: Cell wall proteins were separated after lysis and collected by the 2% ‎SDS (Sodium Dodecyl Sulfate) method. SDS-PAGE (Sodium Dodecyl Sulfate-Poly ‎Acrylamide Gel electrophoresis‏(‏‎ and Dice coefficient were used to evaluate the electrophoretic ‎pattern and percent homology respectively. Cell wall proteins were obtained by the Glass Bead ‎Friction method. Conclusion: The highest level of homology in cell wall proteins was between nail and mouth, ‎toes and arms, sample standard C.albicans and inguinal area with 95% homology and the lowest ‎level of homology was between toes and nail with 55% homology.‎ The highest similar molecular weight in cell wall proteins of all isolates (35 and 64Kd) in 12 ‎specimen, (24 Kd) in 10 specimen and (50 and 26 Kd) in 9 specimen and the lowest similar ‎molecular weight in cell wall proteins (29, 42, 60 Kd) was observed in 8 specimen and (29 Kd) in ‎one specimen of C. albicans. Results: Results obtained from the total protein patterns of cell walls can be used for comparing ‎with patterns cytoplasmic proteins. These results also can be useful for studying in vitro ‎conditions and molecular studies.

Objective: The rate determination of cellular proliferation and cell viability is critical to the assessment of the effects of drugs on cells and there are different standard methods in evaluation of cell proliferation. Calprotectin is a cytosol metaloprotein in the neutrophil, macrophase, lymphocyte and monocyte cytosol that induce growth-inhibitory against various cell types including tumor cells. These finding indicate it can be as a therapeutic agtent in cancer diseases. Biological characteristics of calprotectin are contained antibacterial, antifungal, immonoregulation, chemotactic activity and inhibition of cell proliferation. In the present study, two staining assays; trypan blue and MTT in vitro evaluation of human calprotectin proliferation inhibition on human gastric cancer cells was compared. Materials and Methods: Calprotectin was purified from human neutrophil by chromatography methods. The human gastric adenocarcinoma cell line (AGS) was used. These cells were incubated in RPMI 1640 medium supplemented with 10% FBS in a humidified incubator (37oC & 5% CO2). AGS cells (10000 cell per well) were incubated to different concentration of calprotectin (25, 50, 100, 200, 400 mg/ml) at 24, 48, 72 h. In this study trypan blue dye exclusion and MTT assay were used for evaluation of cell proliferation inhibition.Results: Results of cytotoxicity effect of calprotectin on AGS cells show that there are significant correlation between two methods (P<0.01) and LC50 value of calprotectin calculated by MTT assay is lower than trypan blue in all time intervals. The LC50 of calprotectin for AGS cell at 24, 48 and 72 h was determined 33.29, 71 and 141.8 mg/ml by trypan blue and 96.78, 38.66 and 9.86 mg/ml by MTT assay, respectively. There are significant correlation between both methods at different time intervals and it is positive linear (P<0.01–P<0.001). Conclusion: Result of cell cyto toxicity of calprotectin by both methods indicated that correlation is positive and significant. It can be said that MTT assay is more sensitive, easy to handle, a large number of probes can be assayed in a relatively short time.

Abstract‎ Objective: Aim of this study was to purify Human serum paraoxonase-1, which is one of the ‎important organophosphate detoxifying enzymes. Materials and Methods: In this study we found anion exchange DEAE Sephadex A-50 and gel ‎filtration media Sephadex G-200 is suitable for purification of this enzyme. Using Triton X-100 ‎a non ionic detergent, enzyme was separated from lipoproteins. Then enzyme solution was ‎applied to DEAE column and washed to elute non specific binding. The bind enzyme was eluted ‎with sodium chloride gradient. Active fractions were pooled and applied to G-200 column. The ‎active fractions were applied to the second DEAE column. Changing the buffer and the pH ‎resulted to a purified enzyme. Results: Results of these chromatographic procedures showed purified enzyme with greater than ‎‎95% purity and 320 U/L of specific activity. SDS-Poly acryl amid gel electrophoresis showed a ‎single band of approximately 43 KD protein. Kinetic properties of the purified enzyme showed ‎that, affinity of the enzyme to the substrates is dependent on buffer composition and the calcium ‎and sodium ions concentration. Using paraoxon as the substrate the activity was related to the ‎calcium and sodium concentration (Km=1.39±0.52), but for phenyl acetate as the substrate the ‎activity was independent of sodium and calcium (Km=0.728±0.105). The optimal pH for ‎paraoxon hydrolysis was around 9.5-11, but for phenyl acetate it was from 8-8.5. Glycerol 20% ‎‎(v/v) was most effective stabilizer. Keeping at 25oC for 20 days 75% of the original activity was ‎restored. Conversely at 56oC the ammonium sulfate was better stabilizer. EDTA and zinc ‎chloride both inhibited the enzyme activity. Concomitant incubation of the inhibitors with ‎calcium ion resulted to increased IC50 values.‎ Conclusion: The stablished purification procedure is simple and non-expensive and can be used ‎for large scale purification of this enzyme which can be used for organophosphate ‎decontamination or prevention of intoxication.

Objective: Asthma is a common chronic disease with episodic attacks which induces heavy economic burden on the health system, while by considering an effective and cheap therapeutic method, it is possible to reduce the burden. Salmeterol spray is one of the common used drugs in asthmatic patients. There are three brands of this drug in Iran: Iran-manufactured Salmeterol (Salmeterol), India-manufactured (Serobide) and England-manufactured (Serovent), which have different efficacy and cost and have been studied in this survey.Materials and Methods: In this randomized clinical trial 30 asthmatic patients with inclusion criteria randomly divide into three groups and each group received one of the brands. A base Spirometery was done for each patient firstly, and then they used 50 mcg of drug in two doses (q12h) for three days. For every patient diurnal pick flow meter, three times a day, and also a final Spirometery in the third day of study was done. To calculate cost-efficacy the Economical Preferability Index was used. Results: All three brands caused increase in lung volume, after three days treatment, while Serovent and Salmeterol led to significant rise (P<0.05). The final cost of Iran-manufactured salmeterol to increase one liter in lung volume was less than Serobide but non-significantly while it was significantly less than Serovent. (P<0.05) Conclusion: In recent studies combination therapy of long-acting inhaled bronchodilators like Salmeterol and inhaled corticosteroids is the most effective therapeutic method to control asthma. Regarding numerous asthmatic patients and heavy burden of import of foreign brands and beside considerable difference between their costs, such these studies are valuable due to efficacy and cost-efficacy of Iran-manufactured drugs.

Objective: Chronic viral hepatitis is a common disease that has variable Prevalence in the world. In our country about 2 millions are carrier of HBV and about 200-300 thousands of them have chronic liver disease. Psychological symptoms as depression in the patients are related to: chronicity of the disease, drugs such as Interferon used in the treatment and the fear of complications. We decided to study the prevalence of depression in these patients. Materials and Methods: Our study is a cross-sectional study that done in Tehran hepatitis center. Two questionnaires used in this study. The first included demographic information and progressing of the disease and another was Beck questionnaire. Beck test is a standard questionnaire for evaluation of depression. Result: In the current study 290 patients were evaluated. Average age was 37.4±11.4 years (Min=18 years and Max=69 years). From all of the patients 226 (77.9%) were male and 46 (22.1%) were female. The average of Beck score in men was 10.5±9.2 and in women 14.6±10.5 that this difference was statistically significant (p<0.002). The rate of depression in the patients treated with interferon (INF) was 86.7%, in patients treated with INF-Ribaverin was 59.5%, in patients with no treatment was 37.6 and in patients treated with lamivudine was 35.5% and treatment by interferon had significant difference with all of the groups (p<0.01). Conclusion: According to high prevalence of depression in the patients under treatment with INF, it seems that psychological assessment and treatment is necessary.

Objective: Hemorrhoidal disease is a common anorectal condition which surgical hemorrhoidectomy remains its standard treatment. Post-operative pain is a common and important problem following hemorrhoidectomy. Doppler guided-hemorrhoidal artery ligation (DG-HAL) is a relatively new method which is based on selective ligation of the arteries supplying blood to the piles. In this study we compared the post-operative pain and patients satisfication of conventional hemorrhoidectomy with DG-HAL.  Materials and Methods: This is a radomized clinical trail which carried out on 51 consecutive cases which were admited to Baghiyatallah and Day surgical day clinic from 2004 to 2006. Patients randomized into two groups: group A (n=25) were treated with conventional hemorrhoidectomy and group B (n=26) with Doppler-guided hemorrhoid artery ligation. Post-operative pain assessed by visual analog scale (VAS) and scored between 0-10. Data analyzed using SPSS 11.5 for windows and p values < 0.05 considered statistically significant. Results: Among 51 studied cases, 25 cases treated with DG-HAL method and 26 cases treated by conventional surgery. The average age of patients was 42.29 years and 82.4% were male and 17.6% were female. The average pain scores of patent in the 1st, 2nd and 3rd post-operative day and 1st and 2nd week post operatively were respectively 1.92, 1.08, 0.36, 0.04 and 0.04 for DG-HAL group and 6.58, 6.31, 5.81, 4.58, 2.85 for surgery group. 80% of cases in DG-HAL group returned to their usual daily activities in first 3 days after intervention and in surgery group 15.4% asumed their usual activites in the same period (P value<0.05). The rate of satisfaction in terms of complete satisfaction in HAL group versus conventional hemorrhoidectomy was 76% and 50% respectively; relative satisfaction was 20% versus 38.9% and no satisfaction 4% versus 7.7%.Conclusion: In comparison with conventional hemorroidectomy, post operative pain in HAL technique was less than conventional hemorrhoidectomy and patients satisfication in later tecnique was beter. Return to usual daily activities in HAL group was also considerabley shorter than conventional hemorrhoidectomy.

Objective: Abdominal operations is associated with celiac adhesions; this study compares the frequency and severity of adhesions following cesarean sections performed with and without closure of the parietal and visceral peritoneum.Materials and Methods: A prospective cohort study of women undergoing first repeat cesarean delivery was designed. All surgeons were asked immediately after surgery to score the severity and location of adhesions. Patient records were then abstracted to assess prior surgical technique, including parietal peritoneal closure, other attributes of first surgery, and patient characteristics. The chi square test and multivariable logistic regression were used for statistical comparison and analysis. Results: One hundred fifty patients were enrolled. (70 patients with Prior parietal peritoneal nonclosure and 80 patients with Prior parietal peritoneal closure): closure of the parietal peritoneum at the time of the first cesarean section was associated with significantly less visceral adhesions than closure (45% versus 77%, P=0.006) and significantly fewer dense adhesions (33% versus 48%, P=0.03). Conclusion: Parietal peritoneal closure at primary cesarean delivery was associated with significantly fewer and filmy adhesions. The practice of nonclosure of the parietal peritoneum at cesarean delivery should be questioned.

Objective: Stress urinary incontinence (SUI) is most common form of female Incontinence. The purpose with this study was to determine frequency of SUI and examine obstetric risk factors in Baghiatallah Hospital. Materials and Methods: In a cross- sectional study 400 women assessed by a questionnaire that focusing on clinical finding of SUI in Baghiatallah Hospital in 2005. Other health variables collected from obstetric charts and analyzed.  Results: The frequency of SUI four months after delivery was 4%. In these women, method of delivery (cesarean section versus normal vaginal delivery) was not associated with SUI (CS, 2.7% versus NVD, 5.6%, p=0.33). SUI four month after delivery was associated with younger maternal age (age<25, p=0.04) and higher body mass index (BMI>25, p=0.04(. Conclusion: The frequency of SUI four months after delivery was low in our study. SUI was associated with younger maternal age (<25) and higher BMI (>25) and was not associated with mode of delivery.