Effect of Selenium on Expression of Apoptosis-Related Genes in Cryomedia of Mice Ovary after Vitrification.

INTRODUCTION: Freezing of ovarian tissue is used for preservation of fertility. The freezing-thawing process is accompanied by oxidative stress and induction of apoptosis. Apoptosis is a complex process that has been studied in animal models. The present study was aimed at investigating the effect of selenium on suppression of apoptosis during vitrification-thawing process of mice ovary via studying expression of apoptosis-related genes, and also, we aimed to design statistical models for the roles of single genes and gene-gene interactions in suppression of apoptosis. METHODS: A total of 10 right ovary samples from 10 mice were randomly divided into two groups of selenium treatment (at dose 5 µg/ml sodium selenite, through adding to the media) and control group. Vitrification-thawing process was done according to the existed protocols. Real-time PCR was used for gene expression study. The apoptosis gene profile included P53, Bax, Fas, and Bcl-2. General linear model was applied to study single gene associations and gene-gene interactions. RESULTS: From the studied genes, P53 showed a significant downregulation in the selenium group in comparison to the control group (∆∆CT = 1.96; P = 0.013; relative expression (RE) = 0.28). Bcl-2 showed a significant upregulation in the selenium group in comparison to the control group (∆∆CT = -2.49; P < 0.001; RE = 3.49). No significant result was found for other genes. According to the multiple models, Bcl-2 showed a protective single gene association (beta = -0.33; P = 0.032), and Fas∗Bcl-2 interaction was significantly positive (beta = 0.19; P = 0.036). CONCLUSION: Addition of selenium to cryomedia of vitrification-thawing process could reduce the apoptosis induced by freezing-thawing stress in mice ovary via downregulation of P53 and upregulation of Bcl-2 at transcription level. Multivariable statistical models should be performed in future researches to study biological systems.

Therapeutic effects of curcumin and ursodexycholic acid on non-alcoholic fatty liver disease.

Fatty liver disease is commonly associated with inflammation, oxidative stress and apoptosis of hepatocytes. This study was designed to investigate the combinational therapeutic effects of curcumin (CMN) and Ursodeoxycholic acid (UDCA) on non-alcoholic fatty liver disease (NAFLD). Male Wistar rats were divided into 8 groups: NAFLD-induced rats, NAFLD-induced rats + CMN, NAFLD-induced rats + UDCA, and NAFLD-induced rats that received CMN + UDCA. CMN (200 mg/kg) and UDCA (80 mg/kg) was administered orally for 14 and 28 consecutive days. Biochemical and histopathological analysis were conducted in all the groups. It was seen that co-administration of CMN and UDCA significantly reduced fatty degeneration, cellular necrosis, edema, and immune cell infiltration compared to non-treated NAFLD-induced rats. Whereas, combinational therapy caused a significant decrease in levels of SGOT and SGPT enzymes and expression of p53, caspase III, iNOS and bcl-2 mRNA and proteins, in variant with the treatment of CMN and UDCA, respectively. Co-administration of CMN and UDCA was also associated with the restoration of the levels of serum TG and HDL-C however, had no effect on LDL-C. It also resulted in an in TAC, GSH- PX, and SOD and decrease in MDA level. Our study concludes that combinational therapy of CMN and UDCA is effective for the treatment of NAFLD, as compared to their solo treatment.

Using CuO nanoparticles and hyperthermia in radiotherapy of MCF-7 cell line: synergistic effect in cancer therapy.

The aim of this paper was examining the combined impacts of CuO nanoparticles (CuO NPs), hyperthermia (H), and irradiation (R) on an increment of MCF-7 cells. The MTT assay was employed to assess the antiproliferative effects of CuO NPs (25, 50, and 100 µg/ml), hyperthermia (41 °C for 1 h), and irradiation (200 cGy). Moreover, the perniciousness was estimated through the survival capability of cells, and apoptosis, ROS production, and levels of caspase-3, -8 and -9 proteins were determined. A significant (p < .01) decrease in proliferation index (0.124 ± 0.021), a significant (p < .01) increase in apoptosis (42% ± 1.54) of MCF7 cells, a significant (p < .03) increase in ROS formation (32.16 ± 1.9) and a significant (p < .01) increase in LDH release (33.28 ± 1.56) were recorded in the adjacency of MCF-7 cells by a combination of CuO NPs (100 µg/ml) and R + H compared to control and other treatments. The activities of caspase-3 (0.33 ± 0.014) and caspase-9 (0.389 ± 0.019) also increased significantly (p < .05). However, caspase-8 showed no significant changes in its activity (p = .065). Based on these observations, a combination of CuO NPs, hyperthermia, and irradiation could suppress the growth of MCF-7 cells and evoke cell apoptosis via mitochondrial membrane potential.

Extract of Berula angustifolia (L.) Mertens Enhances Wound Healing in Streptozotocin-induced Diabetic Rats.

INTRODUCTION: Diabetes-impaired wound healing and other tissue abnormalities are considered to be a major concern. OBJECTIVE: The aim of this study is to assess the wound healing activity of the methanolic extracts of Berula angustifolia leaves. MATERIALS AND METHODS: Seven- week-old male Wistar rats with diabetes induced by streptozotocin injection were randomized into 5 groups of 6 rats based on allocated treatment. Wounds were created by an excision-based or incision-based wound model. For wound healing activity, the extracts were applied topically in the form of ointment and compared with the control groups. The healing of the wound was assessed based on excision, incision, hydroxyproline estimation, biomechanical, and biochemical studies. RESULTS: The healing rate of the extract-treated groups was significantly different compared with the control group (P < .05). Hydroxyproline contents increased significantly in the extract-treated groups (P < .05). There were significant differences in the extract-treated versus nonextract-treated groups, particularly in terms of cellular infiltration, acute hemorrhage, congestion, edema, collagen production and density, reepithelialization, and neovascularization. CONCLUSIONS: The methanolic extract of B angustifolia enhances wound healing activity significantly in both studied wound models. From this animal study, enhanced wound contraction, decreased epithelialization time, increased hydroxyproline content, improved mechanical indices, histological characteristics, and biochemical studies suggest the extract of B angustifolia leaves may have therapeutic benefits in diabetes-impaired wound healing.

DNA-Hybridization Detection on Si(100) Surfaces Using Light-Activated Electrochemistry: A Comparative Study between Bovine Serum Albumin and Hexaethylene Glycol as Antifouling Layers.

Light can be used to spatially resolve electrochemical measurements on a semiconductor electrode. This phenomenon has been explored to detect DNA hybridization with light-addressable potentiometric sensors and, more recently, with light-addressable amperometric sensors based on organic-monolayer-protected Si(100). Here, a contribution to the field is presented by comparing sensing performances when bovine serum albumin (BSA) and hexaethylene glycol (OEG6) are employed as antifouling layers that resist nonspecific adsorption to the DNA-modified interface on Si(100) devices. What is observed is that both sensors based on BSA or OEG6 initially allow electrochemical distinction among complementary, noncomplementary, and mismatched DNA targets. However, only surfaces based on OEG6 can sustain electroactivity over time. Our results suggest that this relates to accelerated SiO x formation occasioned by BSA proteins adsorbing on monolayer-protected Si(100) surfaces. Therefore, DNA biosensors were analytically explored on low-doped Si(100) electrodes modified on the molecular level with OEG6 as an antifouling layer. First, light-activated electrochemical responses were recorded over a range of complementary DNA target concentrations. A linear semilog relation was obtained from 1.0 × 10-11 to 1.0 × 10-6 mol L-1 with a correlation coefficient of 0.942. Then, measurements with three independent surfaces indicated a relative standard deviation of 4.5%. Finally, selectivity tests were successfully performed in complex samples consisting of a cocktail mixture of four different DNA sequences. Together, these results indicate that reliable and stable light-activated amperometric DNA sensors can be achieved on Si(100) by employing OEG6 as an antifouling layer.

Assessment of synergistic effect of combining hyperthermia with irradiation and calcium carbonate nanoparticles on proliferation of human breast adenocarcinoma cell line (MCF-7 cells).

The present study was undertaken to evaluate the synergistic effect of combining hyperthermia with irradiation and calcium carbonate nanoparticles (CC NPs) on proliferation of MCF-7 cells. The cells were randomly allocated to 19 groups: one negative control, three positive controls and 15 treatment groups. MCF-7 cells were treated with three concentrations of CC NPs (50, 100 and 150 µg/mL), gamma radiation (200 cGy), hyperthermia (41 °C for 1 h) and three concentrations of doxorubicin (200, 400 and 800 nm) and incubated at 37 °C for 24 h. Then the cell viability, the percentage of apoptosis and the levels of caspase-3, -8 and -9 proteins were measured. The results indicated that the combination group (150 µg/mL CC NPs + thermoradiotherapy) had a significant (p < .001) decrease in cell viability (48.65 ± 4.8%) and a significant (p < .001) increase in apoptosis percentage (45 ± 1.63%) of MCF-7 cells, as compared with the negative control and most of the other treatment groups. Moreover, a significant (p < .05) increase was observed in the activity of caspase-3 and caspase-9. Our findings revealed that CC NPs in combination with irradiation and hyperthermia could significantly reduce the cell viability and enhance the apoptosis of the MCF-7 breast cancer cells, the same as doxorubicin anti-cancer drug.

The Extract of Lycium depressum Stocks Enhances Wound Healing in Streptozotocin-Induced Diabetic Rats.

In diabetes, impaired wound healing and other tissue abnormalities are considered major concerns. The aim of the present study was to assess the wound-healing activity of methanolic extracts of the extract of Lycium depressum leaves. A total of 60 healthy male Wistar diabetic rats weighing approximately 160 to 180 g and 7 weeks of age were randomized into 10 groups for incision and excision wound models: sham surgery group (SHAM), including creation of wounds and no treatment; base formulation group (FG) with creation of wounds and application of base formulation ointment; treatment group 1 (TG1) with 1 g of powder extract of the plant material in ointment; treatment group 2 (TG2) with 2 g; and treatment group 4 (TG3) with 4 g of powder extract of the plant material in ointment. A wound was induced by an excision- and incision-based wound model in male rats. The mature green leaves of L depressum were collected and authenticated. Extractions of dried leaves were carried out. For wound-healing activity, the extracts were applied topically in the form of ointment and compared with control groups. The healing of the wound was assessed based on excision, incision, hydroxyproline estimation, and biomechanical and biochemical studies. The extract of L depressum leaves enhanced wound contraction, decreased epithelialization time, increased hydroxyproline content, and improved mechanical indices and histological characteristics in treatment groups compared with SHAM and FG ( P < .05). These findings permit the conclusion the extract of L depressum benefits parameters of wound healing in a diabetes induced model.