Cell Journal (Yakhteh)
Year:2004 | Volume:6 | Issue:22|Papers Count:8

Introduction: Verapamil, a phenylalkylamin –type ca 2+ channel blocker, is widely used in the treatment of cardiovascular diseases . Material and Methods: In this study , the chronic effects of oral administration of the drug on hematologi and immunologic parameters of male Wistar rats have been studied. Verapamil was administered orally at dosages of 10,20,50 mg/kg for two months, while control rats received only distilled water. We studied leukocyte and erythrocyte counts by use of light microscopy. Results: The lymphocyte counts were decreased (%10) in rats treated with 20,50 mg/kg of verapamil compared to control group. The monocyte counts were signifigantly increased (%5-8) at the end of treatment in the verapamil group. There were no significant differences in the body weight, total white and red blood cell counts ,and neutrophil count compared to control group. Conclusion: These results should be taken into consideration in the chronic treatment of patients with verapamil and they suggest that immune parameters of such patients should be monitored during the treatment course.

Introduction: This Study was designed to compare the effects of three vitrification procedures [conventional (C), open pulled straw (OPS) and closed pulled straw (CPS)] on morphological survival of two-cell mouse embryos and their subsequent development to hatched blastocysts. Material and Methods: Two-cell mouse embryos were divided into four groups, control (194), OPS (150), CPS (155) and C (160). The control embryos were cultured in HTF, for 96-120hrs. Embryos in vitrification group were vitrified with 5.5M EG and 1M sucrose by three different methods. After rapidly thawing at room temperature, embryos were diluted in stepwise sucrose solution (0.5, 0.25 and 0.125). The embryos were rinsed 3 times in HTF methods, then cultured. Survival of embryos in each procedure was determined after thawing. Ability of the survived embryos to continue development was assessed upon subsequent culture in vitro and compared with control group. All of the results were analyzed by chi-square test. Results: The survival rate of the embryos after thawing was significantly higher in CPS (76%) compared to OPS(62%) (P<0.01) and C(66%), (P<0.05). The rate of hatched blastocyst didn’t differ significantly for C (58%), OPS (55%) and CPS (64%), but was significantly lower than control (72%) for C(P<0.05) and OPS (P<0.01). Conclusion: This Study revealed that the CPS method seems to be a usful procedure for cryopreservation of two-cell mouse embryo.

Introduction: It has been shown that γ-IFN is one of the major cytokines involved in lung fibrosis. It has also been demonstrated that Nitric Oxide (NO) may exert vasodilation, edema and cytotoxicity, and it may stimulate cytokinedependent processes in the lungs. In this study, the effect of 17-beta Estradiol (17βE2) and five alpha dihydrotestosterone (5α-DHT) on NO release by TGF beta treated alveolar macrophages was investigated. Material and Methods: Alveolar macrophages were obtained by bronco alveolar lavage of healthy male rats aged 8 to 10 weeks in the usual manner. Lavaged cells (95% alveolar macrophages) were plated out at 1x105 cells/well using a 24 well flat bottomed cell culture plate. Cells were incubated for 2hrs at 37°C and 5% CO2. Non adherent cells were removed by three washings and alveolar macrophages were covered with 1 ml complete medium without phenol Red containing 10 µg/ml LPS,100U γ-IFN and different concentration of 17β E2 or 5α-DHT in the presence or absence of TGF beta. Cells were incubated for another 24h and supernatants were collected and Nitrite was measured using indicator of NO by Griess method. Results: Results showed that TGF-β reduced 5α-DHT induced NO release. The minimum and maximum reduction in NO release was seen in response to 1x10-10M to 1x10-6M, indicating that TGF-β was able to reduced 5α-DHT induced NO. It also showed that 17β E2 reduced NO release in TGF-β treated macrophages in a dose dependent fashion. The minimum and maximum inhibition in NO release was seen in response to 1x10-11M and 1x10-6M respectively. Conclusion: According to results it seems that the part of pulmonary inflammation which is related to TGF-β probably due to nitric oxide release and that sex hormones may play a role in this process.

Introduction: The most important virulence factor of Brucella is related to its capability of intraphagocytic survival. Brucella with this ability,causes persistant infection in host. Garlic extract, because of has organosulfur compounds such as allicin, has antimicrobial activity. The aim of this study was to investigate the effect of garlic choloroformic extract on intramacrophage Brucella spp. Material and Methods: In this study, garlic choloroform was extracted and the quantity of allicin calculated with HPLC; then effect of the extract on intramacrophage survival of B.melitensis Rev1 and B.abortus S19 was studied on cell culture of mouse peritoneal macrophages. Results: Results indicated that the extract was effective and eradicated intramacrophage Brucella in 1:40 (equal to 439 microgram per ml allicin), 1:80 (equal to 218 microgram per ml allicin) and 1:160 (equal to 128 microgram per ml allicin) extract dilutions after 24 hours. Conclusion: Attention must be paid to antimicrobial effect of garlic on intramacrophage Brucella which, seems to be useful in treatment of brucellosis.

Introduction: Survivin, an inhibitor of apoptosis (IAP) containing one baculoviral IAP repeat (BIR) domain, has been reported to be able to regulate both cellular proliferation and apoptotic cell death. In the present study, we evaluated the differential expression of different variants of survivin during prenatal and postnatal development of brain in mice. Material and Methods A total of 27 NMRI mice were categorized into 9 age groups and brain specimens were obtained accordingly (n=3 per each group): 11 and 17 days embryos and newborn from which the remaining brains were collected by intervals of 5 days up to 1 month of age. Total RNA was extracted from each brain and Reverse Transcription was performed by oligo dT and M-MLV enzyme. cDNAs were amplified with primers specific for survivin and ß2 microglobulin (as an internal control) via polymerase chain reaction technique. Results: We demonstrated that survivin is expressed during both fetal and postnatal development of brain in mice. RT-PCR performed on survivin showed two different variants of survivin with different intensities. The expression of the bigger variant (survivin140) during both prenatal development and at birth was significantly higher than its postnatal one. Conclusion: Our data suggests that the expression of survivin140 in brain is developmentally regulated; such a regulation may play a role in homeostasis of brain, and in refinement of synapses.

Introduction: Enrichment of intact peripheral blood dendritic cells (DCs) has been done using rosette and immunomagnetic depletion techniques. Material and Methods: Peripheral blood mononuclear cells were separated using ficoll density gradient centrifugation and T lymphocytes were depleted through reaction with AET- treated sheep red blood cells. The remaining cells were depleted from lineage marker positive cells using monoclonal antibodies against CD3, CD11b, CD14, CD16, CD19, CD56 and anti- mouse IgG magnetic beads. Flowcytometric analysis was used to determine the purity of the obtained DCs through HLA-DR expression and lack of lineage markers. Results: The obtained results showed that a significant population of isolated cells (58.5% ± 4.5, n=5) fail to react with anti-lineage marker monoclonal antibodies but express the HLA-DR antigen. Conclusion Immunomagnetic depletion is a new method for DCs separation that unlike classic methods, can isolate DCs in intact form. According to our information, this is the first report of DCs separation in Iran and purity of the obtained DCs is comparable with similar studies in other countries. Significant enrichment of intact DCs could facilitate the study of DCs natural functions in different fields such as autoimmunity, cancers and infectious diseases.

Introduction: Cyclosporin A (CYA) is a potent immunosuppressive drug widely used in transplant settings due to its specific inhibition of T–cell activation. Several Studies have reported the synergistic effect of CyA with hematopoietic growth factor in colony culture of mice bone marrow. We Studied the effect of IL-3, IL-6, SCF,CyA and nutrition medium ( condition medium - PHA) on the development of hematopoietic cells of human bone marrow because this unique cyclic endecapeptide has been successfully utilized in organ transplantation for graft rejection and graft versus host disease and also since CYA stimulates colony formation by hematopoietic cells in vitro. Material and Methods: Human bone marrow cells were taken from the posterior illiac crest of volunteer donors aged 5-35 years. Cells were cultured in complete culture medium (Containing 12.5% FCS 12.5% horse serum and 50 µmol Hydrocortisone with IL-3, IL-6, SCF and CY-A) on 24 –well microplates at 37 degree centigrade with 5% co2 for 4 week and colony culture was performed for 14 days in semisolid Agar medium .The cell count by light microscope and colony assay by inverted microscope were performed weekly for four weeks. Results: Long term bone marrow culture and semisolid agar culture stimulation with IL-3, IL-6 , SCF and CyA had two as much cloning efficiencies than that of parallel cultures without CyA and IL-3, IL-6 ,SCF in inducing proliferation and conservation of CFU – GM in Bone marrow culture. Culture inactive PHA- condition medium showed higher stimulation than active condition medium. Cloning efficiencies were calculated by mean mean colony number per medium and data were presented as comparison of mean colonies in CyA group to control group. Conclusion: Statistical analysis using the t-student test was performed to determine significance of differences between cultures in presence or absence of CyA. Consequently,these results show a direct positive effect of Cy A on the signal transduction pathways in hematopoietic stem and progenitor cells.

Introduction: Human Embryonic Stem (hES) Cells are pluripotent cells derived form the inner cell mass of blastocyst. These unique cells have the potential to form virtually any cell type in the body and can be propagated in vitro indefinitely in an undifferentiated state. This study was initiated to assess the effect of retinoic acid (RA) on hES cells with and without rosette morphology (neuroectodermal – like cells). Material and Methods: Embryoid bodies (EBs) were made from ES cell line (Royan H1) with and without rosset compartments. The EBs treated with retinoic acid (RA) (10 µM) for 20 days and the cultured on tissue culture plate individually containing neuronal medium. Differentiated cells were evaluated by phase contrast microscopy and immunocytochemistry. Results: Differentiated cells were positive with the neuron specific antibodies against microtubules associated protein 2 (a + b) (MAP – 2) , neurofilament protein (NP), neuron – specific enolase (NSE), synaptophysin, and tubulin III. In presence of RA, more EBs were differentiated into neurons in both groups (in group with rosette compartment, 73.2% v.s. 13.2%, P<0.01, and in group with non – rosette compartment, 57.5% v.s. 19.5% P< 0.01). Moreover, RA in EBs with rosette compartment increased neural morphology. Conclusion: These results showed RA induces hES into neurogenesis and rosette compartments f hES have more potential for neurogenesis and these cells may be used in stem cell transplantation therapies for diseases in future.