JOURNAL OF CELL & TISSUE
Year:2018 | Volume:9 | Issue:3|Papers Count:8

Aim: In this study, the pattern of gene expression (PR1, NCED and SPR2) was considered as informative markers of systemic resistance, and positive beta-amino-butyric acid (BABA) effects was investigated on induction of resistance in Lycopersicon esclentum Mill. Material and Methods: L. esclentum (Tomato) cv. Hungarian was selected in the four-leaved stage. For each pot, 70 ml of a 250 mM BABA solution was prepared and sprayed on plant leaves. The treated pots were kept under controlled conditions (16 h light at 30 ° C and 8 h darkness at 25 ° C) for 2 days, before the bacteria were induced. After two days, the bacteria were inoculated. The total RNA from leaves was extracted at 0, 24, 72, and 96 h after inoculation. The cDNA was synthesized and the gene expression pattern was determined by RT-PCR method. Results: The level of expression of three defense-related genes (PR1, NCED and SPR2) increases as a result of bacterial contamination. However, pre-treatment with BABA resulted in a significant increase of resistance gene expression in response to the challenge of pathogen relative to the control plant and the apparent symptoms of the disease (As roundish and irregular spots up to brown and black with chlorosis) causing damage to tomatoes. Conclusion: Pre-treatment of the plant with BABA has enhanced the plant's defense system, and has shown the extreme effect of BABA on plant resistance. As a result, this indictor can be used to control a P. syringae pv. syring in tomatoes.

Aims: This study was aimed to investigate the anti-proliferative effects of hydroalcoholic extract of Blepharis persica seed and its synergy effect with doxorubicin on human breast cancer and prostate cancer cell lines. Materials and Methods: hydroalcoholic extract of seed was prepared using maceration, and ethanol%70. Eight concentrations of extract and four concentrations of combined with doxorubicin (125, 62. 5ngr/ml) and extract (0. 625, 0. 315mg/ml) were prepared. MCF7, LNCaP and SKM cell lines were cultured. Cell viability was evaluated by MTT assay after 24h. To show apoptosis induction of breast and prostate cancer cell death by extracts, Annexin/PI test were performed. BCL2 gene expression was analyzed using Real-Time RT-qPCR for 24, 48h. Results: The highest effects of growth inhibition were observed in the prostate, breast and fibroblast cell lines with extract, respectively. The combined effect of doxorubicin with extract in three cell lines in comparison with control did not show any significant difference. The results of Annexin / PI indicated that the percentage of initial apoptosis, delayed apoptosis and necrosis in treated cells increased compared to control. BCL2 expression in cell lines decreased significantly over 24 and 48 h compares to control(p<0. 01). Conclusion: hydroalcoholic extract of B. persica seed has ability to inhibit the proliferation of cancer cell lines as well as the induction of apoptosis in cancer cells, compared with using it with doxorubicin, although the extract did not have any synergy effect with doxorubicin at lower concentrations, it can be a substituted for this kind of drug with fewer side effects.

Aim: The aim of this study is to evaluate the interaction effect of salinity stress and superabsorbent polymers on physiological traits of basil. Material and methods: A pot experiment was conducted as factorial based on a completely randomized design with four levels of salinity (0, 40, 80 and 120 mM NaCl in irrigation water) and four levels of superabsorbent polymers included (control, Ackoasorb, Stockosorb and Terracottem). The measured traits were soluble protein, antioxidant enzyme activities, malondialdehyde content and essential oil production. Results: Antioxidant enzyme activities of catalase, guaiacol peroxidase, ascorbate peroxidase and polyphenol oxidase at first harvesting and high salinity (120 mM) under Ackoasorb, Terracottem, Terracottem and Ackoasorb usage decreased 52. 68, 73. 1, 68. 07 and 75. 35%, respectively, and also at second harvest at highest salinity level (80 mM) the activity of these enzymes under Ackoasorb, Terracottem, Ackoasorb, Terracottem and Terracottem decreased 37. 6, 62. 5, 46. 38 43. 06 and 38. 47%, respectively. With increasing salinity the essential oil production decreased and Tracheotem superabsorbent increase it. At first harvesting, a significant positive correlation was observed between malondialdehyde with guaiacol peroxidase (r = 0. 751) and at second harvesting ascorbate peroxidase with protein (r =-0. 753) had a significant negative correlations, moreover, significant positive correlations were found between superoxide dismutase with guaiacol peroxidase (r = 0. 848), malondialdehyde with guaiacol peroxidase (r = 0. 789) and malondialdehyde with ascorbate peroxidase (r = 0. 743). Conclusion: The results showed that application of superabsorbents under salinity stress conditions could decrease the severity of this stress and thereby cause to decrease the antioxidant enzyme activities and malondialdehyde amount significantly, but increased the soluble protein and essential oil content.

Aim: In this research, design and fabrication of graphene reinforced nano-composite scaffolds for neural induction in dental pulp stem cells (DPSCs) has been considered. Material and methods: Polycaprolactone / nano-graphene composite scaffolds with 1%, 3% and 5% wt. graphene were firstly fabricated by the solvent casting method. Subsequently, hydrophilicity and electrical conductivity of the nano-composites were measured. According to the results of the mentioned physical experiments, the appropriate scaffold with the optimal ratio of nano-graphene was selected and cellular morphology, metabolic activity and neural differentiation potential of cultured DPSCs on it were evaluated by scanning electron microscopy (SEM), MTS assay and Immunofluorescent staining, respectively. Results: Conductivity results demonstrated that by adding graphene to pure polymer, the electrical conductivity of the composite considerably increased. The addition of 5% wt. nano-graphene to the polymer can also reduce contact angle amount from 99. 89± 2. 86° to 64. 03± 3. 36° . Finally, SEM and Immunofluorescence images with MAP2 marker illustrated that the cultured cells on the surface of optimum scaffold differentiated into neuron-like cells with neural morphology. Conclusion: The results of this study showed that conductive polycaprolactone containing 5% graphene nano-composite scaffolds have the appropriate potential for induction of neural differentiation and application in neural tissue engineering.

Aim: In the present study, the effects of homocysteinylated and normal forms of substance P were investigated on the proliferation of PC12 cell-line in the presence of cysteine. Material and methods: Cells were cultured in an RPMI-Glutamax medium containing 10% FBS and metabolic activity was assessed in the presence of different concentrations of peptide and cysteine using the MTT assay. Afterward, the morphological changes were determined using phase-contrast microscopy. In order to evaluate the protective effect of homocysteinylated peptide against H2O2, cells were treated with 150 μ g/ml H2O2 for 3 h, and their nitric oxide production was assessed using the Griess colorimetric assay. Results: findings showed that two forms of substance P, at selected concentrations, caused a significant increase in proliferation of PC12 cells and a decrease in the amount of NO production in response to H2O2 toxicity. At a concentration of 6-10 M, homocysteinylated and normal forms of substance P increased the rate of PC12 proliferation up to 78. 65% and 38. 55 % and decreased the levels of NO up to 36% and 14%, respectively. This indicates that homocysteinylated form of substance P is more potent (~40%) than the normal form. The percentage of protection for homocysteinylated substance P was about 22% higher than the normal form. Conclusion: It seems that homocysteinylated substance P can increase the growth rate of cancer cells and promote the disease course through the reduction in NO production in pathological conditions.

Aim: The main goal of this study is to determine optimum conditions for mouse 2-celled and 4-celled single blastomeres for high-qualified development into a blastocyst and the production of embryonic stem cells (ESCs) from these blastocysts without the presence of feeder cells. Material and methods: At first step, 2 and 4-celled embryos were collected from oviducts. Blastomeres were isolated and cultured in 1 and 5μ l of medium in the conditions single, group and with intact embryos. Then, the resultant blastocysts were cultured on gelatin-coated dishes for ESCs production. The expression of specific markers for both blastocysts and ESCs were analyzed with immunocytochemistry and PCR. Results: The results revealed that the volume 1μ l was better than 5μ l and co-culture with embryos and group culture significantly better supported blastocyst development than single culture. Based on Oct4 expression in the ICM of blastocysts, cell count was performed and also showed significant increase in blastocyst quality. We also demonstrated that 4-celled blastomeres not only have heterogeneity in the potential of development among them, but also have lower potential than 2-celled blastomeres. Finally, it was also shown that, single culture derived-blastocysts could generate embryonic stem cells in the specific medium and these cells showed the differentiation potential into different cells. Conclusion: The method of the present study for the evaluation of single blastomeres developmental potential and the production of ESCs can be used for other species. The production of ESCs without feeder cells in human is an important and big challenge.

Aim: In the present study, we examined the possibility of astaxanthin usage in the MS model of mice, experimental autoimmune encephalomyelitis (EAE). Material and methods: EAE disease was induced using MOG35-55 peptide and complete Freund adjuvant (CFU) in female C57BL/6 mice. Mice were divided into control, patient group (MS model, EAE), and prevention group, the group received astaxanthin for 21days before immunization, and astaxanthin treatment was initiated when mice showed the first symptoms of EAE. Each group consisted of 8 mice. The consumption of Astraxanthin was continued for 35 days, and each mouse received astaxanthin about 0. 4% of the weight of used pellet (about 400 mg). Cell proliferation was measured by MTT, IL1β , but IL6 levels (proinflammatory cytokines) were measured by ELISA. In addition, the spinal cord tissues were also taken for immunohistopathological evaluations. Results: Based on the results, the use of astaxanthin produced appropriate clinical and pathological outcomes, lead to decrement of IL1β , and IL6 proinflammatory cytokines (P<0. 0001) production. Immunohistopathologic studies also showed a decrease in leukocyte infiltration in the spinal cord of experimental groups. Conclusion: Overall, this drug approach can be considered as a useful strategy in the prevention and treatment of multiple sclerosis.

Aim: 7-bromoindirubin-3'-oxime (7-BIO) is a well-known glycogen synthase kinase-3β (GSK3β ) inhibitor with anticancer effects on different cancer cell lines. However, due to its low water solubility, absorption and high gastrointestinal toxicity, it has been inapplicable in clinics so far. In the present study, bovine serum albumin nanoparticles (BSA NPs) were prepared as drug carriers. Material and methods: to be conjugated with 7-BIO to increase the effectiveness and reduce the toxicity. The nanoparticles were synthesized and conjugated with 7-BIO and their sizes were analysed by using scanning electron microscopy. Also, the viability and apoptosis in the cells treated with 7-BIO conjugated nanoparticles were examined by MTT assay and acridine orange/ethidium bromide staining (AO/EB), respectively. The significance of the data variation was evaluated by using ANOVA test. Results: 10 mg/ml albumin led to spherical nanoparticles (NPs) of 89. 42± 7 nm in diameter with mono dispersity. MTT assay showed that 7Bio-loaded BSA NPs had a higher toxic effect on MCF-7 cell line compared to the free 7-BIO. Also, AO/EB staining showed that the apoptosis was induced by 7-BIO treatments. 7-BIO is known to specifically inhibit GSK3β phosphorylate GSK3β , which was also evident in our treated breast cancer cell line MCF-7 both with free and loaded BSA NPs 7Bio. Conclusion: The albumin nanoparticles conjugated with the GSK3 inhibitor effectively decreased the viability of the proliferating breast cancer cell line MCF-7, pointing at its possible clinical impact in future.