Study of Morphology and Biochemistry of Rat Bone Marrow Mesenchymal Stem Cells Before and After Osteogenic Differentiation: A Comparative Study
Pages 103-111
https://doi.org/10.52547/JCT.3.2.103
Abstract Aim: The aim of this study was to compare the morphological and biochemical character of mesenchymal stem cells and osteoblasts in vitro.
Material and Methods: After extraction of rat bone marrow mesenchymal stem cells and culturing them till the 3rd passage, these cells were cultured in media containing beta-glycerol phosphate, dexamethasone and ascorbic acid for 21 days. Then morphology of the mesenchymal cells and differentiated one were investigated using Hoechst and acridine orange. In addition the level of matrix deposition, level of calcium, activity of alkaline phosphatase in mesenchymal cells and differentiated on as well as metabolic activity of osteoblasts with the help of MTT was determined.
Results: Morphological study showed delocalization of nuclei and roundness of cytoplasm in differentiated cells as compared to mesenchymal stem cells. In addition mineralization of the matrix started from the 10th day and reached the maximum level at the 21st day. Also from the day 10 the level of calcium increased significantly (p < 0.05) but activity of alkaline phosphatase decreased significantly (p < 0.05) in differentiated cells. Where as during the differentiation process the metabolic activity of the cells increased significantly (p < 0.05).
Conclusion: In addition to morphological differences, calcium content, alkaline phosphatase activity and metabolic activity of the mesenchymal cells and osteoblasts also differs. Therefore in addition to alizarin red staining these factors also can be used to investigate the differentiation processes in vitro.
Curcumin Inhibits the Expression of Aquaporin 5: The New Perspective in Inhibition of Colon Carcinogenesis
Pages 113-117
https://doi.org/10.52547/JCT.3.2.113
Abstract Aim: Expression of aquaporin5 (AQP5), a member of aquaporins family of water channel proteins, in the early stage of colon cancer suggests that AQP5 is probably a driving force in colon carcinogenesis. We hypothesized that curcumin can reduce the level of AQP5 protein in hct116 colon cancer cells.
Material and methods: hct116 colorectal cancer cells were cultured in DMEM, divided into three groups, nontreated control group, vehicle-treated and drug-treated. Cells were just exposed to medium, vehicle-treated (sham) cells received ethanol (as the solution of curcumin) and drug-treated cells were treated with different concentrations (10, 20, 30, 50, 80, 160 µM) of curcumin for 24 and 48h. Cell viability measured by MTT assay. Then cells were treated with 50 µM )IC50) of curcumin. immunocytochemistry performed to examine the effect of curcumin on the expression of AQP5.
Results: MTT assay indicated that curcumin decreased cell viability and proliferation in a time and dose dependent manner.Immunocytochemistry showed that the amount of AQP5 protein decreased in treated cell by curcumin.
Conclusion: our results suggested that curcumin inhibits the expression of AQP5 in human hct116 colon cancer cell line. In this article, for the first time, the effect of curcumin in inhibition of AQP5 investigated. Inhibition of AQP5 expression may provide a novel therapeutic target in colon cancer treatment and prevention.
Comparison of Phenotype of Lymphocytes in ABO Blood Groups
Pages 121-125
https://doi.org/10.52547/JCT.3.2.121
Abstract Aim: Researchers have made considerable efforts to determine the significance of particular ABO antigens to diseases susceptibility such as cancer and infections. O blood group people have a higher risk than other blood groups for catching cholera. Thus relationship between ABO blood groups and catching disease is unknown remained. The aim of this study is to determine the lymphocyte phenotypic profile in ABO blood groups.
Material and Methods: Peripheral blood samples collected from forty health male people with different ABO blood groups (each group=10). All of studied people were in 18-25 years old ranges and had a closely similar genetic background. The samples were studied using a FACSort flow cytometr for determination of CD3+T-lymphocytes phenotype and their subsets CD4+, CD8+ and Treg cells (CD4+/CD25+/Foxp3+), CD19+ B-lymphocytes and their subsets CD5+ and CD5‾ B cells.
Results: There were no significant differences in the frequency of examined lymphocyte subpopulations (CD8+ and Treg lymphocytes, and B- lymphocyte subsets) between the ABO blood groups. But, the CD4+ T lymphocytes percentage in B blood group was higher than other groups (p < /em>=0.05).
Conclusion: High frequency of CD4+T lymphocytes in B blood group may be reduced catching some disease such as bacterial and parasitic infections.
Revelation of Replicative Capability of Hair Follicle Germinative Epidermal Cells after Repeated Plucking: Implications for Stem Cells and Hair Growth Cycle Control
Pages 127-140
https://doi.org/10.52547/JCT.3.2.127
Abstract Aim: The aim of this study was to evaluate the validity of bulge hypothesis on the limited proliferative potential of epidermal cells located at the basal end of hair follicle.
Material and methods: Fibers of rat vibrissa follicles were plucked once or repeatedly and then pattern of cell replication and sequence of regenerative events was examined by measuring length of growing fiber, histology of regenerating follicle and immunohistochemistry (BrdU uptake of epidermal cells).
Results: Data provided here demonstrated that after single or multiple depletions, a new epidermal matrix originated merely from the residual germinative epidermal cells of the follicle base and cells from the bulge region had no role in formation of this structure. The matrix is the source of a new fiber which grows at a equivalent rate to its native counterparts from unplucked follicles. The total length of hair produced from the plucked follicles was much longer when compared to the fiber produced in normal follicles.
Conclusion: Based on our results, replicative potential of epidermal cells located at the follicle base is not limited to a single cycle. We therefore consider it unlikely as cessation of growth phase of follicle is due to restricted proliferative potential of these cells.
Effect of Salt Stress on PSII Efficiency of Dunaliella bardawil under Light and Dark Conditions
Pages 141-151
https://doi.org/10.52547/JCT.3.2.141
Abstract Aim: In this study, the effect of salt stress on PSII of Dunaliella bardawil as a photosynthesis species model was invesigated to determines more effects of this stress and appropriate solutions to salt stress.
Material and methods: In this study, OJIP-test method was used for analyzing of chlorophyll a fluorescence under stress. Different concentrations of NaCl, 1 to 2 M and 1 to 3 M, were applied at light and dark conditions.
Results: The results showed in both light and dark regimes, the rate of Fv/Fo, ΦPo, ψo , ΦEo, ΦRo and PIABS were reduced by salt stress of 1 to 3 M NaCl at primary hours after salt stress while Φ Do was increased. After first hours, the efficiency of PSII did not increased in dark regime unlike the samples in light condition.
Conclusion: According to the results, salt stress led to reduction of water-splitting complex activity and the function of other electron acceptor in PSII. The rate of electron transport to pheophytin, QA, QB and the other electron acceptors is also reduced. Hence it could be finalized that water-splitting complex is the first site damaged by salt stress. On the other hand, studying on salt stress under light and dark regimes showed that light accompanied by the other mechanisms such as photosynthesis and chlorophyll production increase the efficiency of algae system and recover to the condition before stress.
The Concomitant Effect of Shikonin and Glutathione Peroxidase-1 on Enhanced Survival of Dopaminergic Neurons against Parkinsonian Toxicity
Pages 153-160
https://doi.org/10.52547/JCT.3.2.153
Abstract Aim: the aim of this study was to examine the effect of glutathione peroxidase-1 (GPX-1) and shikonin on enhanced survival of dopaminergic neurons PC12 against parkinsonian toxicity.
Material and methods: in order to overexpress GPX-1 in PC12 neurons, recombinant lentiviruses carrying both GPX-1 and reporter GFP genes were generated and used to infect target cells. The survival rate of the transduced neurons in the presence or absence of shikonin was then quantified.
Results: following GFP gene expression observed under the fluorescent microscope, the overexpression of GPX-1 was determined using the RT-PCR analysis. Changes in cell survival against parkinsonian toxicity were examined in the presence of two factors: GPX-1 overexpression and shikonin treatment. The results indicated that both GPX-1 overexpression and shikonin treatment of the PC12 cells increased significantly cell survival against parkinsonian toxicity. Survival increased by 14% after GPX-1 overexpression and by 11% following shikonin treatment. More importantly, when the two factors were applied simultaneously (by shikonin treatment of GPX-1-overexpressing cells) they saved 83% of the neuronal cells that was up by 29%. This increase of survival rate was significant compared to the increase achieved by each factor alone.
Conclusion: our data showed that GPX-1 gene overexpression and shikonin treatment not only individually increase PC12 cell survival against oxidative stress caused by the parkinonian toxin 6-OHDA, but also will function additively and/ or synergistically if they are applied together.
Evaluation of Auxin (IAA) and Kinetin Effects on Lead Uptake and Accumulation in Matthiola flavida Bioss
Pages 161-169
https://doi.org/10.52547/JCT.3.2.161
Abstract Aim: The aim of present study was evaluation of IAA and kinetin effects on lead uptake and accumulation in Matthiola flavida.
Material and methods: Metallicolous population of M. flavida was grown in hydroponic condition. Then plants were exposed to combination of 1µM Pb and 0, 1, 10, 100 µM IAA and kinetin for 14 days. Finally, Pb concentration was determined in their aerial parts and roots using a flam atomic absorption. Also their Chlorophyll contents were measured and calculated by spectrophotometer.
Results: Results showed increasing aerial part Pb concentration more than 48 and 110% respectively in 1 and 10 µM IAA comparing to control. A significant decreasing aerial part Pb concentration was observed with increasing of kinetin concentration in culture medium. Root Pb concentration significantly decreased with increasing of auxin and kinetin in nutrient solution. Significant differences in aerial part dry weight were not observed in different IAA and kinetin concentrations in comprising with control.
Conclusion: Increasing aerial part Pb concentration in M. flavida was observed in low concentration of IAA but kinetin did not show clear and positive effects on biomass and Pb concentration.
Comparison of Sperm Parameters and DNA Integrity between Fertile and Varicocele Individuals
Pages 171-177
https://doi.org/10.52547/JCT.3.2.171
Abstract Aim: Varicocele is the most common causes of male sterility that is caused progressive testicular damage over time. In this study sperm parameters and their DNA integrity was compared in varicocele and fertile people.
Material and Methods: The study performed on 55 grades II and III Varicocele people and 25 fertiles. Semen parameters (sperm count, motility and morphology) and sperm DNA health were assessed using TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling). Data were analyzed by SPSS software.
Result: Varicoceles semen parameters consist of sperm count, motility and morphology had significantly low quality in comparison with fertile people. Also damaged DNA sperms percentage of Varicocele people (14.34%±1.43) was significantly increased compared to fertile people (9.2%±1.36). Sperm parameters and DNA damage percentage correlation showed that there is a reversed correlation between DNA damage percentage and sperm density and significant and positive correlations with sperm abnormal morphology.
Conclusion: Testicular temperature increasing in Varicocele people, caused to low quality of sperm parameters and also high sperm DNA damages. Therefore, Varicocele people are confronted with spermatogenesis process deficiency.
