Volume & Issue: Volume 3, Issue 4, Spring 2013 
Research - Scientific

The Anti- Proliferative and Lethal Effects of D-alpha Tocopheryl Succinate (Vitamin E Succinate) and Honey Bee Venom(BV) on Human PromyelocyteLeukemia Cell Line (HL-60)

Pages 287-296

https://doi.org/10.52547/JCT.3.4.287

Abstract Aim: Acute promyelocytic leukemia is the most common leukemia among adults. Nowadays vitamin E has been used for cancer treatment in research and clinical approach. Application of high concentrations of vitamin E to induce apoptosis have side effects so in this study we examined the effect of bee venom or vitamin E or both together on the induction of apoptosis in HL-60 cell line in order to increase anti-proliferative ability and reduce side effect of vitamin E.
Material and Methods:HL-60 cell line were cultured in RPMI medium and then treated with different concentrations of vitamin E and bee venom. The effect of bee venom and vitamin E on cell viability and proliferation were measured by MTT assay and cell counting trypan blue staining. Data were analyzed using one-way ANOVA test and Instate 3 software.
Result: Our results show that inhibitory effect of vitamin E increased dramatically in the presence of bee venom. On this basis we suggest that non- toxic concentration of bee venom can increase anti –proliferative activity of vitamin E on HL-60 cancer cells.
Conclusion: Our result show that  non-toxic concentration of BV can increase inhibitory effects of vitamin E and it make hope that BV can increases the anti-proliferative  potential of vitamins  in non-toxicconcentration and  concentration  withoutside effectsin the future.
 
 

Research - Scientific

Stereological Study of the Effect of Vitamin E on Rat Kidney Tissue Treated with Para-Nonylphenol

Pages 297-306

https://doi.org/10.52547/JCT.3.4.297

Abstract Aim: Para-Nonylphenol is an environmental pollutant which can generate free radicals causing tissue damage. The aim of this study was to investigate the effect of vitamin E as a strong antioxidant on kidney tissue in rats treated with para-Nonylphenolusing’stereological technique.
Material and Methods: Twenty fourmaleWistar rats with the mean body weight of 198±20 g were randomly divided into 4 groups (n=6), control, vitamin E (100mg/kg/day), Para-Nonylphenol (250mg/kg/day) and Para-Nonylphenol+ vitamin E, and treated orally for 56 days. Finally, the rats were weighted and their right kidneys were taken out, fixed, sectioned, processed and stained using Heidenhain̕& azan method. The kidney tissue was then studied using stereological methods. Data were analyzed using one way ANOVA and means difference was considered significant at p < 0.05.
Results: The mean kidney weight, the total volume of kidney, the volume of cortex and medulla, the volume of proximal and distal tubule lumen, interstitial tissue and the volume of glomerulus and tuft significantly increased in Para-Nonylphenol group compared to the control ones, while the volume of proximal and distal tubules epithelium and the volume of Bowman’s space reduced significantly.
Conclusion: Vitamin E could compensate many adverse effects of Para-Nonylphenol on the kidney tissue. Consequently, vitamin E could be used as a potential antioxidant (supplement) against NP intoxication.
 

Research - Scientific

Cloning and Study the BioinfomaticTrait of TropinoneReductase-II (TR II) Gene fromHyoscyamusniger

Pages 307-318

https://doi.org/10.52547/JCT.3.4.307

Abstract Aim: The purpose of present research was extraction and cloning of tropinonereductase-II gene (tr-II) at antisense direction in pBI121 binary vector to provide transgenic plants with low rate of tropinonereductase-II enzyme and high production of scopolamine and hyoscyamine for future projects.
Material and methods:  Total RNA was extracted from Iranian native Hyoscyamusnigerroots, and the interest gene after cDNA synthesis and cloning at antisense direction in pBI121 binary vector, was transfered to Agrobacterium tumefacience. Accurate cloning was studied through 3 methods; enzymatic digestion, PCR and DNA sequencing. The bioinformatic characters of the gene were then surveyed.
Results: Three used methods confirmed true cloning in high efficiency. Nucleotide sequence of the gene revealed the 783 bp in length, encoding a polypeptide of 260 amino acid residues, with high similarity to that one registered in NCBI. The predicted molecular mass and isoelectric point of deduced polypeptide were 28437.3 Da and 5.46, respectively. Protein structures were not completely similar to those previously reported at PDB data base. Also, phylogenic study demonstrated that this gene belongs to the group I of TRs.
Conclusion: Due to successful cloning and high similarity of nucleotide and polypeptide sequences of gene with those recorded in world data bases; it is expected to get success in access to main purpose.
 
 

Research - Scientific

Application of Seed Storage Protein Marker for Identification of Seven Soybean (Glycine max) Cultivars

Pages 319-327

https://doi.org/10.52547/JCT.3.4.319

Abstract Aim: The purpose of this study is evaluation of seven different varieties of soybean seeds cultivated in Iran based on seed storage protein patterns using SDS-PAGE method.
Material and methods: First, total soluble protein extracted from seeds of seven different varieties of soybean cultivated in Iran including: GK, L17, Sari, 033, 032, Talar and Sahar were measured. The electrophoretic pattern of storage proteins of soybean seeds was evaluated by SDS_PAGE method. Relative density of protein bands was analyzed. Relationships among the seven varieties of soybeans were evaluated based on the presence or absence of protein bands and cluster analysis.
Results: Protein content of soybean seeds showed that total protein in only one variety decreased compared to other varieties. In this study, 11 polymorphic bands on a gel for evaluation of genetic variation were detected and analyzed. Differences in intensities of protein expression were observed among different genotypes. Relationships among the seven varieties of soybeans were studied based on protein patterns and the highest and lowest similarities among varieties were identified.
Conclusion: The protein pattern of soybean seeds based on the presence or absence of protein bands can identify genetic variation and will also introduce specific protein markers that might be used to identify different varieties.
 

Research - Scientific

Regeneration and Somatic Variation in Pelargonium roseum L.

Pages 227-336

https://doi.org/10.52547/JCT.3.4.227

Abstract Aim: The Pelargonium roseum L. is an ornamental plant with important medical properties. The purpose of this study was to optimized the proliferation and regeneration of Pelargonium roseum L. and investigation of the possible somatic variation in the regenerated plant.
Material and methods: In this study first explants from the terminal buds as well as some small part of the stem were prepared and cultured in MS basal medium. Then some fragments of the cultured explants including parts of stems and leaves were isolated and transferred to mediums containing different growth regulators to stimulate micro-regeneration of the plants. From regenerated plants several lines were selected and their morphological characters as well as genetic diversity were evaluated.
Results: The rate of shoots and roots production was maximum in mediums containing leaf explants supplemented with 2mg/l BAP and 0.1 mg/l NAA. Somatic variation of the regenerated lines in comparison with control group showed differences in leaf shape and number, in addition genetic differences were expressed in form of band addition and band removal in bonding pattern of DNA.
Conclusion: The different percentage of shoots and roots production was an indication of the inductive effect of mediums and explants variation on regeneration of the Pelargonium roseum L. Comparison of genetic variation and morphological diversity of regenerated plants could show the common base of genetic for somatic variation.
 
 

Research - Scientific

Function and Phenotype of Lymphocytes in Normal and Inflamed Appendix in Appendicitis Patients

Pages 337-343

https://doi.org/10.52547/JCT.3.4.337

Abstract Aim: Appendicitis is one of the most common abdominal inflammatory diseases. The function of appendix is not clearly defined. In this study to understand the immunological function of the appendix, we investigated the function and phenotypic pattern of lymphocyte in appendix of patients with normal and inflamed appendix tissue.
Material and Methods: Appendix tissue and appendiceal mononuclear cells obtained from 81 patients (mean age; 23±10.5), who was clinicallysuspected of having appendicitis were collected. Based on histopathological examination, twenty-five patients had normal appendix while 40 and 16 were diagnosed with suppurative and gangrene appendicitis, respectively. The phenotypic characteristics of lymphocyte subsets in appendix was analyzed using three color-flow cytometry. In addition, the proliferative response of tissue mononuclear cells was assayed by MTT method.
Results: A significant difference (p < 0. 01) was observed  in the percentage of CD19/HLA-DR, HLA-DR and CD19 cellsfrom patients suffering from suppurative appendicitis in comparison with the patients having normal appendix or gangrene one.  There was also a significant difference (p < 0.01) in the proliferative responses of the appendiceal mononuclear cells to PHA and LPS from pathients with suppurative appendicitis when compare with other groups.
Conclusion: The phenotypic and function of lymphocytes are different between normal and inflamed appendix tissue. These results showed that the appendix tissue with special lymphocyte profile may be effective in preventing or reducing intestinal infections.
 

Research - Scientific

Modeling Breast Cancer Using Chemical and Radiopharmaceutical Effect of P32 on Related Cancer Cells

Pages 345-350

https://doi.org/10.52547/JCT.3.4.345

Abstract Aim: In this article, the ability of p < sup>32 radioactive drugs for treatment and control of breast cancer in rat model and also verifying of distribution of drug in rat organs are investigated.
Material and Methods: 10 to 20 mg per kg of DMBA gavaged to SD strain rats. Tumors 15-10 mm in diameter were selected for testing. P32 radioactive drugs injected to developed tumors with interval 3, 5 and 7 days. The distribution of the P32 radioactive drugs in the body of rat were analyzed by NI detector. After the treatment period, animals were anesthetized and the tumor is completely removed under sterile conditions and placed in 10% formalin. Sections of the tumors were obtained and processed using Tissue Processor. After tissue processing and sectioning, the samples were stained with hematoxylin - eosin were examined.
Results: The results obtained from the examining histological sections prepared from tumor tissue confirmed that, the pathology of tumors induced by DMBA in rats used in this study is very similar to human mammary tumors. Five days after injection, we could find tumor's cells and lymph ducts are covered by residual epithelial cells. Many of the cells are separated from the basement membrane and collapsed into the tubular cavity. Rest of the cells has small nuclei, with chromatin condensation mode and cytoplasm was compact and dense. In radius of about 7-8 mm around the site of injection the tumor's cells were necrosis and formed a cavity, inside of tumor.
Conclusions: These observations showed further penetration and influences of radiation emitted from the phosphor 32 in the tumor tissue based on our injection pattern.
Therefore if given the proper distance of injection within the tumor, destroying the tumor's ability to be on time will be shorter.
 

Research - Scientific

Protective Effects of Ephedra pachyclada on Animal Models with Acute Liver Disease Induced by Carbon Tetrachloride

Pages 351-362

https://doi.org/10.52547/JCT.3.4.351

Abstract Aim: Evaluation ofprotective effectsof Ephedra pachyclada on carbon tetrachloride- induced acuteliverinjury.
Material and method: In this study,initially wedesignedan experiment to achievea standardanimalmodel, in which 30mice were divided into three control, sham andmodelgroups. In themodel group the animal received2ml/kgcarbon tetrachloride50 %( 1:1 in sterile olive oil)intraperitoneally. 72 hourslater,bloodand liversamples were collected and analyzed using biochemicalas well ashistologicalstudies. Afterensuring theeffectiveness of the modelinduction, in the secondtrial, 40 micewere divided into4 groups including controlandthreeexperimentalgroups. Induction ofacuteliverinjury was performedsame asprevious on experimentalgroups, whichtwoofthemweretreatedwith Ephedra pachycladaextract(140, 1400 mg / kg) daily. Bloodand liversampleswere collected96 hafter carbon tetrachloride injection then serumactivity of Alanine amino transferase (ALT), Aspartateamino transferase (AST) andnecrosisas well asinflammationin the liverwere studied.
Result: Treatmentwith Ephedra pachycladaextract significantly decreased the activity of ALTandAST in the serum as well as thenecrosisandinflammation in liver tissue. In addition the survival rate was increased in the animal receivingcarbon tetrachloride.
Conclusion: Results of this researchsuggestthat Ephedra pachycladaprotects the liver against acute liver injuryinduced by carbon tetrachloridethroughinhibition ofoxidative stressandsuppressing theinflammation.