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: The current research was done to investigate the sperm plasma membrane integrity, oxidative stress factors of sperm and testis in male mice treated with cadmium.
Material and Methods: 24 adult NMRI mice were divided into four groups: 1. Control group 2.  Treated with cadmium chloride (5 mg/kg) 3. Treated with Curcumin (100 mg/kg) 4. Treated with curcumin + cadmium chloride. The treatments were performed as a single dose and after 24 hours, epididymal spermatozoa of different groups were used to evaluate the plasma membrane integrity. In addition, the amount of malondialdehyde (MDA) and the anti-oxidant activity of serum and testis were evaluated. Data were analyzed using ANOVA test followed by Turkey’s test (p < 0.05).
Results: findings showed that cadmium chloride caused a significant decrement in plasma membrane integrity compared to the control groups. In addition, cadmium chloride induced a significant increment of MDA in serum and testis and a significant decrement in total antioxidant activity of serum and testis as compared to the group. In curcumin + cadmium chloride- treated group, curcumin could significantly compensate the toxic effect of cadmium on these parameters compared to the cadmium-treated group.
Conclusion: it seems thatcurcumin as a potent antioxidant is able to ameliorate the adverse effects of cadmium chloride on sperm plasma membrane integrity, lipid oxidation and total antioxidant activity of serum and testis.
 

Abstract Aim: The aim of this study was to evaluate the effect of silymarin and lithium chloride on DNA integrity and nucleus of ram sperm.
Material and Methods: In this study, Farahani's ram testes were obtained from Arak slaughterhouse immediately after ram daily slaughter and transferred to the research laboratory. A few incisions were made in the epididymis, and spermatozoa were  then washed into a sterile falcon tube by Ham's F10 medium. collected spermatozoa of ram were divided into four groups: 1. Sperm at 0 hour, 2. Sperm incubated for180 minutes (control), 3. Sperm treated with lithium chloride for 180 minutes and 4. Sperm treated with silymarin + lithium chloride for 180 minutes. DNA integrity and DNA fragmentation were investigated by acridine orange staining sperm chromatin expersion (SCD) test respectively. Morphological feature of apoptosis in sperm nucleus was assessed using Diff-Quick staining. Data were analyzed using one-way analysis of variance test (ANOVA) followed by Turkey's test .
Results: The percentage of DNA fragment and apoptosis were significantly increased in lithium chloride-treated group compared to the control. In silymarin+ lithium chloride group, Silymarin could signicantly compensate these effect compared to the lithium choloride group.
Conclusion: Silymarin as a potent antioxidant could  prevent toxic effect of  lithium  on DNA  fragmentation and apoptosis in sperm nucleus.