Cell and Tissue Journal

Abstract Aim: In order to study the efficiency of the tuber specific promoter (Patatin1), the expression of the HBsAg antigen of the hepatitis B vaccine was evaluated using a transient expression method (AgroInfiltration) in the tubers and leaves of potato plants.
Material and Methods: A tuber specific promoter (Pat1) was isolated from the potato plant using a specific primer pairs by Polymerase Chain Reaction (PCR). It was cloned in the upstream of a synthetic optimized codon of the HBsAg gene in the binary plant vector pBI121. In order to compare the tissue specificity of Pat1, the HBsAg gene also was used under the control of a consultative CaMV35S promoter. The genetic constructs were transferred to the tubers and leaves of potato plants using the Agroinfiltration method. An HBsAg antigen content was measured using ELISA method.
Results: The level of HBsAg antigens in the leaves and tubers of potato plants indicated that Pat1 promoter specifically induced HBsAg high expression in the tuber tissues. Very low expression by the Pat1 promoter in the leaf tissues has been also reported and can be partly dependent on the presence of inducing factors in the inoculation media. However, the expression of HBsAg antigen occurs in both leaf and tuber tissues under the consultative promoter CaMV35S.  The results showed that the codon optimized HBsAg gene for potato plant was expressed properly in plant tissues.
Conclusion:  findings indicate that potato tuber specific promoter Pat1 can be used effectively to express the synthetic optimized HBsAg antigen in the transgenic potato plants.
 
 

Abstract Aim: In the present study, the expression of Beclin1 and LC3 genes has been investigated in 30 patients with non-small cell lung cancer.
Material and methods: RNA was extracted from tumor and adjacent normal tissues of 30 patients with non-small cell lung cancer. After synthesis of cDNA, real time-PCR was exploited for quantitative expression of Beclin1 and LC3 genes. Also, the expression of these genes was compared with the clinical and pathological findings of the patients.
Results: the expression levels of Beclin1 (p < 0.0001) and LC3 (p < 0.0001) genes increased significantly. Also, there was a significant correlation between increasing the expression of LC3 with the subtype of tissue (P=0.01).
Conclusion: The increased expression of Beclin-1 and LC3 genes may enhance the process of autophagy and this condition can develop tumorigenesis; however more research is needed to confirm these findings.
 

Abstract Aim: In this study, the histology effect of caffeine administration was evaluated on retina and PAX6 expression in rat neonates.
Material and Methods: A total of 24 pregnant rats were divided into three groups. Intraperitoneal injection of saline in the control group and also 50 mg/kg and 100 mg/kg caffeine injection from the ninth day of gestation was done. At 20 or 21 embryonic day, neonates on the first and fifth days were euthanized anesthetized and the eyes were isolated and fixed. The neonates were examined for abnormalities. The Retina Histological and morphometric evaluations, eye tissue fluorescence changes, and PAX6 gene expression were investigated by Real time PCR.
Results: The results showed that the effects of maternal caffeine treatment on the retina in neonates are associated with atrophic changes and cell counts of ganglion cell layer, retinal thickness and thickness of the inner plexiform layer, in the animals were treated with caffeine, were significantly reduced (p < 0.05). Also caffeine induced behavioral changes in mothers and tissue fluorescence and significantly increased (P <0.05) the PAX6 gene expression in the groups receiving caffeine.
Conclusion: Administration of caffeine during pregnancy can induce histopathological changes in organogenesis and developing in rat neonate’s retina.
 

Abstract Aim: The aim of this study was to investigate the bovine chondrocytes growth on new polymeric nanofibers silk fibroin/kitosan  scaffolds, designed by authors,  to form the cartilage tissue.
Material and Methods: The chondrocyte cells isolated from three calf articular cartilage were loaded on the   scaffold. After a monthof  incubation, cells morphology was studied by SEM and the rate of cell growth by H&E and DAPI staining. To assay chondrogenesis in the culture, production of collagene was assayed by masson’s trichrom staining and formation of Glycosaminoglycans (GAGs) and  Proteoglycan was assayed by DMMB and “safranin O” staining.
Results: The results showed that the cells attached and proliferated to scaffolds very well. The number of cultured cells after one month proliferated by 4 to 5 times. Masson’s trichrom, safranin O staining and GAG assay showed cartilage tissue production in the scaffolds.
Conclusion: According to the results, the scaffold of nanofibers Fibroin silk / chitosan could be a good scaffold candidate for cartilage tissue engineering. This is due to the nature of the proteins in silk fibroin and polysaccharides in chitosan for cellular attachment and also the diameter close to the diameter of extracellular matrix proteins in these scaffolds.
 

Abstract Aim: In the present study, The anti-metastatic and cytotoxicity effects of cerium oxide nanoparticles on two MCF-7 and T47D breast cancer cell lines have been studied.
Material and methods: MCF-7 and T47D breast cell lines and normal HEK293 cells were treated with 6.5 mg, 650μg, 65μg, 650ng, 65 ng of cerium oxide nanoparticles and MTT analysis was performed to investigate the toxicity of nanoparticles. Then, NM23 and KAI-1 genes expression were measured by real-time PCR method.
Results: Concentrations of 650 μg and 6.5 mg of CNP resulted in approximately 60% death of MCF-7 and T47D cells. Also, in treatment of normal cells with a concentration of 6.5 mg of CNP, Reduced survival of 25% was observed. Gene expression analysis showed that the two anti-metastatic NM23 and KAI-1 genes did not increase expression under  CNP treatment.
Conclusion: According to the results of this study, the cerium oxide nanoparticles have toxic effects on breast cancer cells, However, this effect varies dose and type of cell line dependent. On the other hand, the study of the anti-metastatic expression of genes showed that this nanoparticle is not capable of enhancing the expression of these genes and therefore is ineffective in controlling cell invasion and is likely to exert its anticancer effect through induction of cell death.
 

Abstract Aim: The aim of this study was to evaluate the effect of walnut green skin extract on blue mold apple, peroxidase activity and catalase activity, as well as the level of gene expression of these two enzymes.
Material and methods: In this study, extracts of green skin of walnut were used to control the apple blue mold caused by Penicillium expansum, in vitro and in vivo. Then, the effect of the aqueous extract of this plant on the activity and also expression of the genes of the two enzymes of peroxidase and catalase were evaluated.
Results: The results of the extract mixed with culture media tests showed the aqueous and methanolic extracts of green walnut skin inhibit with the rate of 86.41 and 75.84 percent of the fungal pathogen were respectively the best treatment in comparison with the control. In vivo test (the 4 ° C), the spot surface, in the treatment of aqueous and alcoholic extracts with a concentration of  6×1000, reduced 94.50 and 81.69%, respectively. In the test for the activity of peroxidase and catalase activity, the walnut skin extract increase the activity of these enzymes at 9th day. The results of enzymes genes expression showed that the expression of peroxidase and catalase genes was 227.66 and 314.08 fold on day 9, in the plant extract+ pathogen treatment, respectively.
Conclusion: The extract of walnut skin had direct fungal effects, and it can was induce the defense genes expression and subsequently increased activity of defense enzymes.
 
 

Abstract Aim: The role of Hsp70 chaperone from Rutilus frisii kutum in the thermal inactivation of luciferase in E. coli cell carrying Hsp70 and firefly luciferase was investigated.
Material and Methods: Co-transformation of E. coli cell was carried out with two expression vectors containing Hsp70 and firefly luciferase. The co-transformed cells carrying Hsp70 and luciferase were expressed under optimum conditions. After adding tetracycline, the cells were then incubated for 60 min at 40, 42, 44, 46, 48 and 50°C treatments. Finally, the luminescence activity of samples was calculated.
Results: After 30 min at 40 and 42°C temperatures, the luciferase activity in control samples reached almost zero, while in the co-transformed samples, about 72% and 60% of the luminance activity maintained compared with control samples (before heat treatment), and even after 60 min, up to 36% and 22% of the activity remained. Also, at 44 and 46°C temperatures in the initial times after stress, a significant difference was observed between the luciferase activity of the co-transformed and the control samples. In contrast, at 48 and 50°C temperatures, the changes of the luciferase activity of co-transformed samples were small compared with control samples even in the early stages of stress.
Conclusion: The thermal aggregation of luciferase as a significant reporter protein is inhibited at high temperatures via the activity of Hsp70 in the bacterial cell, which this process can be widely used in the food and pharmaceutical industries and the providing cancer diagnostic kits.
 

Abstract Aim: The purpose of this study was to investigate the effects of benfotiamine treatment of hyperglycemic rats on viability of adipose tissue- derived mesenchymal stem cells on sciatic nerve acellular scaffolds.
Material and method: After induction of hyperglycemia (STZ) and treatment with benfotiamine segments for 4 and 8 weeks from the middle part of the sciatic nerves were  decellularized using Sandell method and seeded with  mesenchymal stem cells derived from adipose tissue. after 8 days of culture cells viability in the culture medium was evaluated by MTT assay and  adhesion of the cells to the scaffold was examined by scanning electron microscopy (SEM).
Results: In comparison with intact group, cell viability of untreated hyperglycemic rats was significantly decreased while, there was no significant difference between benfotiamine  treated group and intact. Results of electron microscopy confirmed adhesion of  the cells on the scaffolds.
Conclusion: In hyperglycemic condition it is likely that glycosylation of ECM constituents and the production of AGEs decrease the inducible effects of ECM on the level of cell viability and probably cell adhesion to the scaffolds. It seems that benfotiamine treatment of hyperglycemic rats by reduction of glycation and AGEs production may prevent the structural changes of the ECM.