Tehran University Medical Journal
Year:2008 | Volume:66 | Issue:4|Papers Count:16

Background: MSCs have been isolated from a variety of mammals by the plastic adherence method. However, this method can be problematic due to the unwanted growth of hematopoietic cells and non-MSCs. The potential of MSCs to differentiate along multiple lineages is the key to the identification of stem cell populations in the absence of molecular markers. In the present study, we describe a homogeneous population of MSCs from mouse bone marrow isolated using an improved plastic adherence method that employs frequent medium change (FMC) at the initial hours of harvested bone marrow cell culture.Methods: Balb/c mice were sacrificed and whole bone marrow cells were aspirated from the femur and tibia and then cultivated in six-well plates. After 3-4 hours of culture, old medium was removed and fresh medium was added. FMC was performed every eight hours over a 72 hour period. When primary cultures became nearly confluent, the first passage was performed. These cells were then used for further examination. To investigate their mesenchymal nature, the cells were allowed to differentiate into mesenchymal lineages and examined at each passage up to the tenth passage for surface antigens by flow cytometry. Results: We achieved purified populations of fibroblast-like cells in the two weeks after culture initiation. The cells were capable of differentiating into osteocytes and adipocytes. Isolated MSCs were reactive to the CD44, Sca-1, and CD90 cell surface markers. MSCs were negative for hematopoietic surface markers such as CD34, CD11b, CD45, CD31, CD106, CD117 and CD135.Conclusions: This protocol provides an efficient isolation of homogeneous populations of MSCs from mouse bone marrow.

Background: Alkaloids are a group of nitrogenous compounds with potential effects on the physiological behavior of human and animals. Some of these compounds are considered important drugs in modern medicine, such as atropine and morphine. Plants are considered the most important source of alkaloids. Therefore, investigating the presence of alkaloids in different plants is very important. Usually, alkaloids in plants are identified by methods such as those of Dragendorf, Wagner and Meyer, among others, which require milligrams of alkaloids for identification. In the present study, a fast and sensitive procedure for detecting of alkaloids in plants is presented.Methods: Twelve dried plants samples were investigated for the presence alkaloids. After extracting the total alkaloid into methanol using a Soxhlet extractor, a few milligrams of the extract was transferred to a separatory funnel, buffered to pH 4.7, the bromocresol green (BCG) solution (10-4 M) was added, mixed and extracted with CHCl3 until a yellow color was observed in the CHCl3 layer, indicating the presence of the alkaloid. The crude extracts were also investigated by the standard methods of Dragendorf, Wagner and Meyer for the presence of alkaloids.Results: Investigation of the 12 plant samples for the presence of alkaloids by the standard reagents of Dragendorf, Wagner, and Meyer showed that only Camelia sinensis (flowers), Echium amoenum Fisch & Mey (flowers), and Stachys (aerial parts) are devoid of alkaloids, with all other samples positive for alkaloids. By the BCG procedure, similar results were obtained, except for the E. amoenum flower, which was positive. The minimum detectable limit for alkaloids by the BCG method is the equivalent of approximately 40mg atropine.Conclusions: According to previous reports, only one of these plants does not contain alkaloids. All studied plants positive for alkaloids by standard reagents were positive by the BCG procedure. Stachys was negative for alkaloids by both the standard reagents and the BCG method, in agreement with previous reports. However, black tea, reported to contain xanthine alkaloids, was negative for alkaloids by both the standard reagents and the BCG method. Therefore, the BCG method is not suitable for the detection of xanthine alkaloids. Nevertheless, the microgram detectable limit for alkaloids indicates that the BCG method is very sensitive.

Background: Ewing’s sarcoma is one of the most malignant tumors in children and young adults. Only a few established cell lines of Ewing’s sarcoma have been reported, which makes it difficult to study the biological features of these tumors. We have recently established a new Ewing’s sarcoma cell line designated SS-ES-1, originating from a thoracic tumor of a 16-year-old female patient. The SS-ES-1 cells have been grown continuously in culture for over 90 passages. In this report, some characteristics of SS-ES-1 cells are presented.Methods: The cells were grown in DMEM medium supplemented with 10% fetal bovine serum (FBS), 100 mg/ml streptomycin and 100 U/ml penicillin in a humidified atmosphere with 7% CO2 at 37 1/4C. The cells were immunocytochemically characterized using a panel of monoclonal and polyclonal antibodies. Furthermore, the chemo-sensitivity of this cell line to some anticancer drugs was assessed using MTT assay and IC50 values were determined. Results: Morphologically, the SS-ES-1 cell line is composed of poorly differentiated small round cells growing in a multilayer pattern. The immunocytochemical staining demonstrates strong reactivity to CD99, cytokeratin, neurofilament, p53 and Ki67 proteins, but no reactivity to GFAP. Based on IC50 values, SS-ES-1 cells display considerable sensitivity to vinblastine (2±0.7 pM), followed by vincristine (0.3±0.12 nM), doxorubicin (0.05±0.03 mM), etoposide (0.64±0.28 mM) and cisplatin (0.67±0.45 mM). Conclusions: In conclusion, the SS-ES-1 cell line demonstrates unique cellular properties, which make it a useful model for studying various aspects of the biology of Ewing’s sarcoma.

Background: The induction of catalytic antibodies (abzymes) was first postulated by Pauling in 1948. Various catalytic antibodies have been detected recently in the sera of patients with several autoimmune pathologies such as systemic lupus erythematosus and rheumatoid arthritis. In addition, antibodies with DNase and RNase activity have been discovered in the milk and sera of healthy human mothers, which shows the physiologic role of these antibodies. In this study, we examined the proteolytic activity of antibodies in the sera of pregnant women.Methods: IgG antibody fractions were isolated from the sera of 30 healthy pregnant women in the first trimester of pregnancy and 10 control samples (men and nonpregnant women) by subsequent steps of chromatographic purification on Protein G sepharose and sephacryl S-300. All patients were in their first pregnancy and aged 25-35 years. The conditions for proteolytic activity, such as type of buffer, pH and temperature, were optimized. The proteolytic activity of these antibodies was demonstrated by in-gel assay with gelatin as the substrate.Results: Antibody treatments at the optimum temperature showed that some samples from pregnant women contain proteolytic abzymes, as demonstrated by in-gel assays. Western blot results confirmed that the proteolytic activity is an intrinsic property of the antibodies. Conclusions: During pregnancy and immediately after delivery women very often experience autoimmune processes similar to those in patients with autoimmune disease. Because of their specific immune status, pregnant women can produce various catalytic antibodies with different enzymatic activity. These proteolytic abzymes might be involved in the direct clearance of antigens from blood.

Background: The Ross procedure has been known as a good method for aortic valve replacement. Pulmonary allograft postoperative stenosis subsequent to the Ross procedure has been noted as an important disadvantage of this technique, although risk factors related to this complication are not clearly recognized. In this study we evaluate the risk factors of pulmonary allograft stenosis after Ross procedure. Methods: This retrospective cohort study was carried out on 42 patients who underwent the Ross procedure. Left ventricular outflow tract obstruction was repaired using pulmonary allografts 22.7±2.5 mm in diameter. Echocardiographic and clinical examinations were performed for all patients at six and 12 months after surgery. Echocardiographic data as well as data related to the allografts were recorded.Results: The mean age of the patients was 28.5±10 years. Postoperative pulmonary valve stenosis was detected in 13 patients (31%). Nine patients (21.4%) had mild stenosis and four patients (9.5%) had moderate stenosis. No relationship was detected between the degree of stenosis and the size of allograft (p=0.08). There was a significant correlation between postoperative pulmonary stenosis and donor age (p=0.04). Predictive variables of pulmonary allograft stenosis after the Ross procedure were low donor and recipient age (p=0.03 and 0.05, respectively).Conclusions: This study shows that the Ross procedure has a rather low incidence of postoperative stenosis and is a suitable substitute for aortic valve replacement. Low donor age is associated with pulmonary allograft stenosis formation, perhaps due to greater viability of tissue cells from younger donors.

Background: The function of internal systems can be influenced significantly by hypernatremia, even anesthesia may be affected by this electrolyte imbalance. The aim of this study was to determine the effects of a single anesthetic dose of ketamine in an experimental rat model of chronic hypernatremia.Methods: Sixty male Wistar rats, weighing 300 (±20)g, were allocated randomly to three groups: the control group received drinking water and test groups 1 and 2 received 1% and 2% NaCl, respectively, for 144 hours. We measured the parameters of sensitivity to the ketamine injection (125mg/kg) including length of times to the inhibition of the righting reflex, inhibition of the response to painful stimulus and the times to the reappearance of the response to painful stimulus and recovery from anesthesia. We also determined the mortality rates during anesthesia. Results: The times for inhibition of the righting reflex and response to painful stimulus for group 2 were significantly shorter than those for group 1 and the control group. The times for the reappearance of response to painful stimulus and full recovery from anesthesia in group 2 were significantly longer than those of group 1 and the control group.Conclusions: Hypernatremia affects ketamine anesthesia in the rat, increasing the speed of passing through the different steps of anesthesia. The duration of ketamine efficacy increases, while recovery from anesthesia is significantly delayed.

Background: The annual amount of mortality in ESRD exceeds the expectation and represents the recent evidences of the inflammation as its etiology. The etiology of inflammation is not clearly known. Chronic inflammation is a dominant occurrence of ESRD which increases the risk of atherosclerosis, malnutrition and peripheral vascular disease. Inflammatory responses are orchestrated by cytokines. Some of the proinflammatory cytokines like IL-6 have a crucial role in this phenomenon. The IL-6 and its receptor activity is up regulated in ESRD patients and the increased level of IL-6 predicts cardiovascular mortality and morbidity in normal and CRF patients. This study devotes itself to determining the serum level of IL-6 and factors affecting it in patients undergoing chronic hemodialysis in Imam Khomeini Hospital which can represent the Iranian Society. By identifying factors affecting the serum level of IL-6 and high-risk patients we can provide treatment possibilities, a decrease in mortality and an improvement in its prognosis.Methods: In this study 42 patients in Imam Dialysis Center were chosen and their serum IL-6 levels were measured at 2 times at three month interval and at the same time blood sample analysis were done for the following: Alb CPR, Ca, P, PTH, TIBC, Ferritin, TG, Chol, LDL, HDL, Uric Acid, Hb, WBC and urea.Results: The mean serum level of IL-6 in hemodialysis patients was 6.35±4.47pg/ml (minimum: 0.55, maximum: 18.25) with the normal range of 1.3±3.2pg/ml. Conclusions: The IL-6 level was higher than normal range in the 52% of the patients. The serum IL-6 level had a significant correlations with CPR, Ferritin, TIBC, WBC and their serum IL-6 level was significantly higher in patients with hypertension, but no significant correlation was observed between other parameters and IL-6.

Background: Hyperglycemia after acute stroke is a common finding that has been associated with an increased risk of death. For the last several years, it was believed that post-stroke hyperglycemia may worsen brain infarction in animal models. According to previous studies, the anti-inflammatory effect of insulin has a protective role on ischemic tissues. Glucose-insulin-potassium (GIK) infusions can be safely administered to acute stroke patients with mild to moderate hyperglycemia producing a physiological but attenuated glucose response to acute stroke, the effectiveness of which remains to be elucidated. In this study, we compared the effects of GIK infusion vs. placebo in ischemic stroke patients.Methods: In this double blind randomized controlled trial, the intervention group consisted of 48 patients who received GIK infusion for 24 hours and the control group included 50 patients who received normal saline infusion for 24 hours. All patients had been admitted to Shariati Hospital during the first 24 hours after the onset of ischemic stroke. At one and three months after the treatment, patients were examined for morbidity and mortality using the Bethel and Modified– Rankin-Scale (MRS) questionnaires.Results: GIK infusions significantly reduced plasma glucose concentrations. Statistically significant differences between the two groups were observed for disability and mortality. However, in spite of the protective effects of insulin and adverse effect of hyperglycemia in ischemic tissues, we observed no significant therapeutic effect from the GIK solution on patient outcome.Conclusions: Although hyperglycemia following acute stroke has been associated with subsequent mortality and impaired neurological recovery, the maintenance of euglycemia in the acute phase has not been shown to improve prognosis. According to this study, GIK infusion has no significant clinical benefit on the outcome of stroke patients. Thus, we cannot recommend routine use of GIK infusion in post-stroke hyperglycemia as an adjuvant treatment for ischemic stroke.

Background: Transfusion-Transmitted Virus (TTV) is a nonenveloped, single-stranded and circular DNA virus belongs to circuviridae family genus Anellovirus, discovered by Nishizawa in 1997. As the usage of common syringes is the known and most common route of the virus transmission, and because of increasing population of Injection drug users (IDUs) we decided to study infection rate in IDU population of our Society. Methods: In a cross-sectional study at Infectious ward of Imam Khomeini hospital, 60 IDU patients were studied. Blood samples were dispatched to lab in citrated test tube for Genome Virus isolation operation, using boiling method, then PCR assay performed based on their available primers. Patient’s information gathered by interview and questionnaire methods.Results: All of our 60 patients were men and their age average was 35.30 (SD±9.68) years old. 26 (43.3%) patients had positive TTV PCR and 24 (92.30%) of them had prison history. 23 (88.50%) of these 26 patients had positive HCV Ab, 17 (65.40%) had positive HIV Ab and 8 (30.80%) had positive HBS Ag. Of 60 study patients 48 (80%) had HCV Ab+, 43(71.70%) HIV Ab+, 26 (43%) TTV PCR and 43 (26.70%) had HBS Ag+ Of 26 patients who had TTV, 34.60% of them had no contemporary sickness and 11.50% of them displayed clear sign of hepatitis (fever, abdominal pain, nausea, vomiting, RUQ tenderness and Icter). 34.60% of them had LFT more than Upper limit normal (45u/l). Time average of injection in 26 TTV patient was 9 years (SD±7.16) and the patient’s age average was 36.35% (SD±9.2).Conclusions: One of the most important route of TTV infection is use of common syringes, TTV infection transmission chance is less than HIV and HCV infection and is more than HBV. In regard to high prevalence of TTV infection in IDU population and because there is no comprehensive information about pathogenesis of this virus in addition to another way of transmission of the virus, the fecal-oral way, we must make plans and policies to decrease danger of transmission of this virus to health care workers, their families and other near relatives.

Background: Much has changed in neonatal care for extremely low birth weight (ELBW; birth weight <1000g) infants over the recent years in Iran, resulting in an increase in their survival rate. We determined neonatal survival and short-term morbidity rates among ELBW infants, and the correlation of these factors with mortality risk. Methods: All single live-born ELBW babies delivered at Vali-e-Asr Hospital over a fouryear period were identified. Data that were prospectively collected included: maternal age, prenatal and perinatal complications, antenatal steroid use, birth weight, gestational age, route of delivery, sex, need for resuscitation, APGAR score, need for mechanical ventilation, and complications including respiratory distress syndrome (RDS), sepsis, jaundice, intraventricular hemorrhage (IVH), necrotizing entrocolitis (NEC) metabolic and hematologic and cardiac disorders. Information was statistically analyzed and a probability value of <0.05 was considered significant.Results: A total of 93 neonates were included in the study. The survival rate in the delivery room was 36.6%. The lowest birth weight was 400g, with a mean birth weight of 850.43 (±136.48)g, and the lowest gestational age was 22 weeks. The mean gestational period was 28.31 (±2.77) weeks. The most frequent complication after birth was RDS (69.89%) and the least frequent was NEC (6.4%). Short and normal gestation was observed in 44.08% and 48.4% of the neonates, respectively. There was a significant relationship between neonatal mortality and the need for resuscitation after birth, RDS and need for a respirator (p<0.05). Furthermore, there was significant increase in mortality among neonates with gestational age <30 weeks, birth weight under 750g and lack of steroids before birth (p<0.05).Conclusions: To enhance perinatal and neonatal care and decrease the rate of preterm labor, improved standard resuscitation and increased administration of antenatal steroids and respirators are required.

Background: Chronic pancreatitis and perivasculitis is the most common etiology of splenic vein thrombosis (SVT). Reported in up to 45% of patients with chronic pancreatitis, SVT may also be seen in patients with acute pancreatitis and pancreatic adenocarcinoma. It causes a localized portal hypertension called sinistral portal hypertension. Unlike those with generalized portal hypertension, patients with sinistral portal hypertension are asymptomatic and have normal liver function. Upper gastrointestinal bleeding from gastric varices is a life threatening complication of SVT. In patients with gastrointestinal bleeding secondary to esophageal or gastric varices, late-phase celiac angiography is used to determine the presence SVT. Splenectomy is effective in treating the collateral outflow for patients with massive gastrointestinal bleeding.Case report: a 23-year-old patient with SVT due to chronic pancreatitis with gastrointestinal bleeding. The patient came to the hospital with upper gastrointestinal bleeding of unknown etiology. Diagnostic workups revealed chronic pancreatitis and SVT with bleeding gastric fundal varices, after which the patient underwent splenectomy. A review of current literature on SVT, known etiologies, diagnosis and treatment is discussed.