مجله سلول و بافت
سال ششم شماره 1 (بهار 1394)

Our aim in this study was to analyze and quantify mRNA expression of SALL4 in mesencephalon, during different stages of chicken embryogenesis. In this experimental study, incubated Ross fertilized eggs were applied in 37°C-37.5°C in 60-65% humidified atmosphere after beginning of embryogenesis. Mesencephalon part of the brain tissue was collected from the eggs, daily. Total RNA extraction and cDNA synthesis was performed from resected tissues. The synthesized cDNA was used as template for quantitatively analysis of SALL4 mRNA expression by real-time PCR. The Real-time PCR analysis of SALL4 mRNA expression in mesencephalon tissues indicated that the level of gene expression is significantly variable during embryogenesis. While the basal level of SALL4 mRNA expression was detected during the rest of embryogenesis, the maximum copy number of SALL4 mRNA was quantified on 19th day of chicken development. Having analyzed the level of SALL4 mRNA expression in different stages of chicken embryogenesis, we can extrapolate that a probable relationship may be existed between expression of SALL4 in nerve centers of mesencephalon brain and development of optic organs.

Periwinkle (Catharanthus roseus) is a medicinal plant as a result of which its medicinal compounds and alkaloids have been under a great consideration. Lead is one of the most common heavy metal contaminant in the environment. Induction of oxidative stress is a result of the presence of heavy metals in plant cells. Due to the problem of heavy metal pollution and the dangers of pollution to plant in this study, the effects of oxidative stress caused by the presence of lead on activity of antioxidant enzymes, level of prolin and alkaloid in callus grown on MS medium with different concentrations of lead were investigated. Catharanthus roseus seeds were planted in the vase containing Pearlite, then to induce callus, the leaves under the sterile condition were put on the solid MS culture media. Calluses were subcultured after three weeks, the calluses after two subculture were treated with different concentrations of lead nitrate (0, 10 and 50 μM). The activity of antioxidant enzymes and content of proline and alkaloid were assayed. Data analyzed at P ≤ 0.05 level using SPSS software. The results of this study showed that the activity of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX), catalase (CAT) and the level of proline and alkaloid as well as lipid peroxidation increased significantly (p<0.05) in comparison to control. Due to induction of oxidative stress, lead was caused an increase in antioxidant enzymes activity and the content of malondialdehyde, alkaloid and proline.

In this study, it was done to evaluate the efficiency of both PRP and Fibrin Glue scaffolds in producing suitable environment for the growth of mesenchymal stem cells. In this study, the preparation of PRP and Fibrin Glue Scaffold were carry out and Mesenchymal Stem Cells (MSCs) isolated from adipose tissue. The mesenchymal phenotype of these cells was determined by mesenchymal surface marker using flow cytometry. Then, MSCs were cultured, and on the third passage (P3)stage, they were seeded separately on the two scaffolds and after 48 hours of cell culture, the ability of the scaffolds seeded cells was evaluated by MTT assay for cells viability. Flow cytometry results showed that human adipose-derived mesenchymal stem cell expressed CD44, CD90 and CD105 surface markers. Also, the results of this study showed that the active PRP could be creating a more suitable environment for the survival and proliferation of mesenchymal stem cells in compare with Fibrin glue It is suggested that the PRP as a protective scaffold could be used for mesenchymal stem cells growth to provide effective strategies in order to tissue engineering development and regenerative medicine.

In this study the hepatoprotectivity effect of Malva neglecta hydroethanolic extract (MHE) were investigated in male rats. 35 male rats with 230-250 gr weight were divided in 5 groups: control (taking normal saline,0.5ml/day, IP), sham(taking olive oil, 0.5ml/day,IP),carbon tetrachloride 1:1 with olive oil,0.5ml single dose, IP), treated 1&2 (carbon tetrachloride 1:1 with olive oil,0.5ml single dose and 300mg/Kg and 600mg/Kg MHE /day for 4 days, IP). After the end of examination the blood samples were collected from heart directly and AST, ALT and AP enzymes were analyzed. The microscopic studies of hepatic tissue were done and histological changes observed by (H&E) staining. All data analyzed with ANOVA test. Statistical significance was accepted at P<0.05. The serum liver enzymes were increased in rats induced by ccl4 compare to the control group (P<0.001) and decreased significantly in rats by treated MHE (P<0.01). Histological finding revealed the necrosis in hepatic tissue. In animals treated with MHE the results showed that the reduction in necrotic cells and increase in hepatocytes regeneration. Our results showed that the ccl4 has hepatotoxicity effect and it can improve inflammation and necrosis in liver tissue. The MHE have antioxidant and flavonoid that can decrease toxic effect of carbon tetrachloride in liver tissue.

The aim of this study was the evaluation of possible genetic and protein pattern changes of callus and of Artemisia plant.

In this study, 2 weeks old shoot buds and shoots grown in MS medium were exposed to gamma radiation with 50,100 and 200 Gy. Then five RAPD_PCR primers, five ISSR-PCR primers for evaluation of somaclonal variation and SDS-PAGE for electrophoresis pattern of proteins in callus of Artemisia plants were studies.

Results showed that the evaluation of DNA bands amplified in the callus and plant samples treated with gamma ray compared to the untreated samples using OPA20 in plant and FPK105 and (GATA)4 in callus in PCR products was obvious but no difference was observed using other primers. In electrophoresis of protein of plant increasing of protein bands due to application of 50 and 200 Gy were observed.

Due to the possible effects by free radicals and reactive oxygen species produced by gamma radiation caused changes in genetic and gene expression.

The purpose of this study was to identify LEAFY homologous genes in a type of nightshade (Solanum villosum Mill.). Total RNA was isolated from shoot apical meristem tissue of S. villosum and was used for First-strand cDNA synthesis. The specific primers were designed based on nucleotide sequence alignment of LEAFY homologous genes from other plants and were used in RT-PCR. The RT-PCR products were purified and sequenced. The results indicated amplification of 540 nucleotides fragment, and named SvLFY. Comparison of SvLFY deduced protein sequences with LEAFY homologous proteins from other species showed high similarity between this fragment and in other C-terminal regions. These results suggest that SvLFY may play a similar role as LEAFY in flower development and could act as a flower meristem identity gene in Solanum villosum Mill.

The aim of this study was to investigate the ability of Lepidium draba L. seedlings to eliminate zinc and silver ions from the medium. The seeds of L. draba were cultured on MS solid medium for 15 days in the presence of different concentrations of both silver and zinc ions. The bioaccumulation of these ions in L. draba tissues were measured by atomic absorption. Furthermore, biochemical and morphological properties of the seedlings such as catalase and superoxide dismutase activities, total flavonoid content, germination percentage, root and stem lengths were analyzed. Results revealed that L. draba seedlings were able to absorb high level of zinc and silver ions and a direct relationship between the level of absorption and the amount of ions in media was seen. On the other hand, total flavonoid contents, catalase and superoxide dismutase activities were elevated by increasing concentrations of the ions in media. Silver ion increased the germination, while the seed germination did not show significant different in treatment with zinc ion in compared to the control. Upon the treatment seedlings with silver ion, the stem growth was significantly increased, while the root growth was decreased. Significant decrease in the stem and root growth were occurred upon seedlings treatment with high level concentrations of zinc ion. Based on the results, it seems that L. draba could be introduced an appropriate option for refining of the aforementioned metals from contaminated area.

Cadmium is one of the air pollutants and a part of diesel exhaust particles. The aim of this study was determination of the effects of cadmium on developmental process and protein pattern of anther and pollen grains in Petunia hybridia that is a common decorative plant in landscapes and parks. Petunia plants were grown in experimental pots and treated with the different concentrations of CdCl2 as spray. Flowers and young buds were collected, fixed in FAA70 (formalin: acetic acid: ethanol with a ratio of 2: 1: 17) and subjected to developmental studies. Pollen grains of different groups were also collected and their proteins were studied using electrophoresis method. Statistical analysis was done using statistical software SAS (version 9.1) and one-way ANOVA analysis. The developmental process of anther and pollen grains in control plants were followed from the pattern of dicotyledonous plants, but some changes and abnormalities were observed in Cd-treated plants. Irregularity in anther wall, formation of wrinkled and vacuolated pollen grains, decreasing of pollen viability, damaged exine and formation of new protein bands are the effect of Cd treatment. Cadmium, as an atmospheric pollutant, influences developmental stages of anther and pollen grains. It causes to form different abnormalities and reduction of pollen viability. Formation of new proteins, detoxifying proteins, is a response to pollution stress.

The aim of this study was to investigate the protective effect of Nigella sativa oil (NSO),as an antioxidant, on the testicular tissue in mice treated with bisphenol A. In this study, 24 adult male mice NMRI were divided into four groups (n=6): control, bisphenol A (200 mg/kg/day), nigella sativa oil (5 ml/kg/day) and bisphenol A + nigella sativa oil. 34 days after oral treatment, the right testis was taken out, fixed and processed followed by Heidenhain's Azan staining. The morphometric parameters of the testicular tissue were evaluated using stereological methods. The mean volume of seminiferous tubules and it's height of the germinal epithelium, the mean diameter and thickness of the basement membrane, the mean number of spermatocytes, round and long spermatids, spermatogonia and sertoli cells significantly reduced in the bisphenol A group compared to the controls (P<0.05). The mean volume of seminiferous tubules and the mean number of Sertoli cells significantly increased in the bisphenol A + nigella sativa oil group to the control level, The mean number of spermatocytes and round and long spermatids also showed a significant increase in this group compared to the bisphenol A group (P<0.05). Co-administration of nigella sativa oil with bisphenol A can prevent the adverse effects of bisphenol A on the testicular tissue in adult mice.

The purpose of this study was enhancement of hGH synthesis by chemical components addition to Serum-Free CHO cells culture and investigate the characteristics of produced hGH.

The effects of different treatments including various concentrations of glycerol, DMSO, NaBu and ZnSO4 in CHO cells were investigated and the GH production was assessed using ELISA method. Cells are seeded into each well of a 12 well plate containing DMEM-F12 and 10% FBS. After 24 hours, media was aspirated and replaced with SFM and treatment with different concentrations of chemicals and rhGH production was assessed using ELISA, dot blotting and western blotting.

Concentrations of 1%DMSO, 0.5% Glycerol, 1mM NaBu and 50 μM ZnSO4 were caused to elevate the expression of rhGH in the CHO cells.

These findings indicate that control of culture aid to development of an efficient large-scale and industrial process for the production of rhGH.