Cell and Tissue Journal

Abstract Aim: Aquaporin4 (AQP4) is the main water channel in the brain that has important role in water transport across blood- brain- barrier. It has been proved that intracranial pressure and also expression of AQP4 increase in some disorders such as Hydrocephalus, Hyponatremia, cytotoxic oedema and brain tumours and drugs that can decrease the expression of this AQP can offer some treatment for these disorders. The aim of this study was to determine whether green tea extract (GTE) can down regulate the Aquaporin4 protein level in brain cortex.
Material and methods: This research is a laboratory experimental study. 40 Wister rats (4-6 weeks) were randomly divided into 4 groups as follows: untreated control group, sham group treated intra peritoneally (IP) with 200 µl saline (solvent GTE), experimental1&2treated IP with 100& 200 mg/kg GTE dissolved in 200 µl saline. After 6 hours, expression of AQP4 in control, sham, and experimental groups were tested by immunohistochemistry and western blotting.
Result: Results from immunohistochemistry and western blotting showed that GTE can down regulate the AQP4 expression level on dose-dependent manner in brain cortex.
Conclusion: It is suggested that GTE has potential to reduce the level of AQP4 protein in brain cortex and it can be useful as a herbal medicine to reduce intracranial pressure in diseases such as hyponatremia, cytotoxic oedema, brain tumours and etc.

Abstract Aim: In this study, the effects of curcumin- coated silver nanoparticles were examined on induction of apoptosis in ovarian cancer A2780 cell.
Material and Methods: Silver nanoparticles coated with curcumin were biosynthesized, then A2780 cells were treated with different concentration of silver nanoparticles. Cytotoxicity effects of silver nanoparticles in A2780 cells were assessed by MTT assay and apoptotic effects of these nanoparticles were examined using DAPI and acridine orange/ propidium iodide staining and caspase 3/9 activation assay. Changes in Bax and Bcl-2 gene expression were analyzed by Real Time PCR.
Results: Findings showed that silver nanoparticles inhibit A2780 cells proliferation in a dose dependent manner. 8 µg/ml 50% decreased cell viability at 24 hours, DAPI and, acridine orange propidium iodide staining represent that percentage of apoptotic cells in the treated groups was increased. The results of Real Time PCR showed that Bax gene expression in cells treated with silver nanoparticles increased, while Bcl-2 gene expression in the treated cells was decreased.
Conclusion: Silver nanoparticles coated with curcumin induce apoptosis in A2780 cancer cells. The use of the nanoparticles should be considered a promising strategy for the treatment of ovarian cancer.

Abstract Aim: The aim of the present study was determination of FecB and FecG^H gene mutations in Kamouri goats of Khouzestan Province. Material and Methods: For this study 40 blood samples were collected of prolific kamouri goats. DNA of blood samples was extracted by modified salting out method and the site of the FecB and GDF9 genes were amplified using specific primers. Results: After amplification of the site of genes, the products of 190 bp and 139 bp were determined by agarose gel electrophoresis. The PCR products were digested with AvaII and DdeI enzymes. Results show no mutations in the FecB and FecG^H genes in the Khouzestan kamouri goats. Conclusion: In this study the mutations of FecB and FecG^H are not observed in Khouzestan kamouri goats and all of the goats were wild type. Therefore these mutations are not the general cause of the prolificacy in the Khouzestan Kamouri goats and further investigations are required to evaluating the fecundity genes and the genotyping of Khouzestan Kamuori goats.

Abstract Aim: In the current study, leaf anatomical structure and tissue cellular variation was evaluated in three populations of M. anisodon in Iran.
Material and Methods: Two individuals were selected from each population. Leaves from middle part of the stems were fixed in F.A.A fixative and thin hand–cut sections were prepared of them.  Then leaf samples were double stained and studied using a light microscope. Data were analysed using SPSS and MVSP softwares.
Results: In all studied populations, leaves trans sections were dorsi–ventral. But, qualitative and quantitative traits of leaf differed among the populations. The ANOVA test did not show significant variations among most of the quantitative traits. Populations clustered separately in the UPGMA tree and also PCA and PCO plots. The population’s arrangement pattern did not fit with geographical distances between populations.
Conclusion: Similarity in the ecological conditions of habitats lead to anatomical structural likeness, and this case dose not related with geographical distance. Therefore, wherever the ecological conditions are similar, the likeness of leaf anatomical structure is very possible.
 

Abstract Aim: The purpose of this study was to investigate the effects of leptin on in vitro maturation, fertilization and embryonic development of immature mouse oocytes.
Material and Methods: Immature oocytes were isolated from NMRI female mice and divided randomly into 4 groups: leptin 0 ng/ml (control), 10 ng/ml, 50 ng/ml, 100 ng/ml. 24 h later matured (MII) oocytes were identified and fertilized with incubated sperm. Embryonic development was investigated every 24 h using inverted microscope.
 Results: From isolated oocytes 53/1% in control group, 56/2% in first group,42/4% in second group and 38/3% in third group developed to MII. The rate of GVBD was 22/05% in control group, 31/27% in first group, 40/63% in second group and 36/39% in third group. The physiologic concentration of leptin (10 ng/ml) considerably increased the rate of MII when compared to other groups (p < 0.05), whereas the GVBD rate was higher in group treated with concentrations more than physiologic dose.
Conclusion:  When different concentration of leptin was applied after IVF, no significant effects were observed between four treatment groups in different stages of embryonic development.
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Abstract Aim: The aim of the present study was to investigate the effects of royal jelly (RJ) and vitamin C (vit C) against PHZ-induced nephropathy in mice.
Material and Methods: Adult male mice were randomly divided into eight eight-membered groups. The PHZ was administered to four groups of mice at a dose of 60 mg/kg per 48 hours intraperitoneally for 35 days. Three of these groups received vit C (250 mg/kg per day) intraperitoneally, RJ (100 mg/kg per day) orally and vit C+RJ with same doses, four hours before PHZ administration, respectively. A vehicle-treated control group and vit C, RJ and vit C+RJ control groups were also included. Serum and renal tissue samples were collected 24 hours after the last treatment and subjected to biochemical and histological examinations, respectively. The data were analyzed by one-way analysis of variance followed by Duncan test for post hoc comparisons.
Results: The PHZ treatment caused a significant (P < 0.05) increased malondealdehyde concentration in serum, but decreased total antioxidant capacity of it. Moreover, PHZ significantly (p < 0.05) increased luminal diameter of proximal convoluted tubule, while decreased (p < 0.05) their epithelial cells height. Considerably, co-treatment of vit C and RJ improved all changes in the above-mentioned parameters.
Conclusion: The Vit C and RJ can minimize PHZ-induced nephropathy in mouse through oxidative reactions inhibition.

Abstract Aim: In this project, production of haploid parthenogenetic embryonic stem cells has been studied in presence of the ERK and TGF-β signaling pathway inhibitors. Material and Methods: First the parthenogenetic blastocyst- produced with chemical activation of oocytes- was cultured in R2i culture condition (containing PD0325901 and SB431542 which inhibit ERK and TGF-β signaling pathways respectively). Then parthenogenetic embryonic stem cell lines was subsequently expanded and preserved in the same culture medium. Using immunofluorescent staining and qRT-PCR, expression of the pluripotent markers were evaluated. Moreover, differentiation potential of these cells was assessed by spontaneous differentiation and teratoma formation assay. Degree of haploid in this cell lines was determined by flowcytometry methods. Results: In R2i culture condition, about 50% of parthenogenetic embryos produce cell lines, out of which 10-13% are haploid. Established parthenogenetic stem cell lines demonstrate pluripotent stemness characteristics including morphology, passagability as well as expression of pluripotent specific genes in mRNA and protein levels. In addition, these cells were able to differentiate spontaneously to embryonic germ layers and form teratoma. Conclusion: Inhibition of ERK and TGF-β signaling pathways can provide an appropriate condition for producing haploid stem cells from parthenogenetic embryos.  

Abstract < p >Aim: Curcumin, a yellow pigment obtained from turmeric (Curcumina longa). It is a dietary polyphenol that has been reportedto possess anticancer, anti-inflammatory, and antioxidant properties. Curcumin is potent radical scavenger and has key role in cell death prevention and because of its antioxidant ability, curcumin has possible role in protection against neurodegenerative diseases. In this study we evaluate the effects of curcumin on rat embryonic neuroprogenitor cells viability and proliferation.
Material and methods: In this study brain cortex of 15.5 day old embryos of Wistar rat dissected out and enzymaticaly dissociated to form single cell suspension. Cell suspension seeded in DMEM/F12 medium supplemented with N2, 1% antibiotic and mitogenes (10 ng.ml^-1EGF and 20 ng.ml^-1 b-FGF). Cells were treated with different concentrations of curcumin (0.1, 0.5, and 1μM). Cell viability was evaluated with MTT assay and immunocytochemical analysis was performed to evaluate astrocytic marker (GFAP). The data were analyzed with One-way ANOVA with Tukey’s post hoc test.
Results: curcumin (0.5 μM) enhanced astrocytes neurite size (p < 0.001) and also has significante effects on neuroprogenitor cells differentiation in culture media.
Conclusion: Data suggest that curcumin enhanced neuroprogenitor cells viability, growth and promoted differentiation of these cells in to astrocytes.
 

Abstract Aim: Dunaliella as a phytoplanktonic unicellular alga is important in aquatic organism nutrition (special in salinity waters). Chlorophyll (chl) content in relation to other algal biomass characteristics was studied under different culture media conditions for better strain selection.
Material and methods: Different Iranian and foreign strains alga were exposed to nitrate deficiency, light, iron and salt increasing treatments. Then some physiological characters such as chls contents, cell number, dry weight, photosynthesis and respiration were measured in control and treated samples.
Results: Different stresses caused decreasing cell number, chls contents and also changing in chla/chlb ratio from 5 to 15. The amount of chla and total chl on a per cell basis (10^-6 cells), at all conditions and the chla amount per ml, only at control showed positive or possitive is correct significant correlation with strains dry weight. Chla or total chl per 10⁶ cells had negative significant correlation with cell numbers per ml in all of conditions.
Conclusion: Algal biomass evaluation that almost is assessed by chla content, do not have a direct relationship with the cell number or dry weight at all cases. Therefore, the higher dry weight, chl content and also cell divisions introduced as better and more suitable indicators for strain selection than others. Strains 10 and 11, having more dry weight at both conditions (control and treatments) nearly with the higher chl content, then 4, 6 and 13 could be introduced.
 

Abstract Aim: In this study, the effect of IAA on salt tolerance of tobacco plants (Nicotiana plumbaginifolia) was investigated under in vitro culture conditions.
Material and Methods: Sterile seeds of N. plumbaginifolia were cultured in MS medium. In vitro propagated seedlings under tissue culture conditions were treated with IAA (mg / l) and salinity (0, 100 and 200 mM) for 4 weeks. Physiological and biochemical parameters including H2O2, chlorophyll, soluble carbohydrate, protein and antioxidant enzyme activity were measured.
Results: Salt stress increased H2O2, proline, catalase, superoxide dismutase and ascorbate peroxidase, while reverse patterns were observed for photosynthetic pigments, soluble carbohydrates and also total protein.  In salt-treated seedlings, application of auxin increased proline, catalase and H2O2, but decreased photosynthetic pigmentation, soluble carbohydrates, total protein and some enzymes such as superoxide dismutase and ascorbate peroxidase.
Conclusion: Auxin treatment in salt-treated tobacco seedlings improved salinity tolerance by changes in some physiological parameters.
 
 

Abstract Angiogenesis is a necessary process in many physiological and pathological events such as tumor growth. Therefore angiogenesis therapy can be a strategy to cancer treatment. It is important to identify different aspects of tumor’s angiogenesis.  Formation of functional vessels within tumors is essential for its growth and progression. The induction of new blood vessel growth by tumors is mediated through different molecules. A balance between pro-angiogenic and anti-angiogenic growth factors strictly control the process. This fact has led to design of therapeutic agents against tumor angiogenesis. Recently, endogenous (such as interleukin -8) and exogenous angiogenesis inhibitors (such as: Avastin) have been described. Furthermore because of the low mutagenic potential of endothelial cells, tumor’s vessel do not show resistance to the effects of many of these compounds. In addition, it is recognized that anti-angiogenesis and anti-cancer tumors can be more effective by combination of factors than the use of either alone. This article would review different aspects of angiogenesis in tumors. Since tumor growth is dependent on the development of blood vessels in the tumor, inhibition of angiogenesis maybe considered an appropriate treatment for cancer. 
 

Abstract Aim: 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.
Material and Method: 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.
Results: 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.
Conclusion: These findings indicate that control of culture aid to development of an efficient large-scale and industrial process for the production of rhGH.

Abstract Aim: Infertility is the failure of a couple to engender after endeavoring at least one year of unprotected intercourse. male factor infertility accounts for approximately 50 percent of causes. Tumor necrosis factor-α (TNFα) is a multifunctional cytokine. TNF-α plays important role in the regulation of cellular processes related to spermatogenesis. There are two variants of the cell receptors that interacts with TNF-α. In the present study, the association of TNFR1 36A/G polymorphisms with idiopathic male infertility in the Guilan population was studied. Materials and Methods: This study consists of 106 infertile men and 114 fertile men as control group. Blood samples were taken and genomic DNA was extracted. Then genotypes and allele frequencies were assessed by PCR-RFLP method and the statistical analysis was performed by MedCalc software. Results: The frequencies of AA, AG, GG genotypes in patients were 41.5%, 7.5% and 50.9%, respectively and in controls were 45.6%, 33.3%, and 21.1%, respectively. The frequencies of A and G in patients were 0.45 and 0.55, respectively and in controls were 0.62 and 0.38, respectively. The results showed that there is a significant association between genotype frequency (p= 0.002) and allele frequency (p= 0.0004) in infertile and control groups. Conclusion: In conclusion, the subjects with G allele and GG genotype appears to be at greater risk of developing idiopathic infertility in Guilan province. Further studies with larger numbers of patients are required to elucidate the potential role of TNFR1 polymorphism in male infertility.

Abstract Aim: During embryonic development, both bird’s feather follicle and mammal’s hair follicle originate by a similar manner, the aim of this research was to access and compare histology, cell growth potential and inductive properties of papilla cells this organs for initiating dermal-epidermal interactions.
Material and Methods: Feather and hair follicles were provided from house pigeons and PVG rats, respectively. Some of the follicles were processed for histological examination whiles others were used to dissect out their dermal papilla. The dissected dermal papillae were cultured in vitro. The cultured cells were then implanted into papilla-depleted follicles. After 28 days the implanted follicles were histologically examined.
Results: Histological surveys revealed that despite some distinct differences, both follicles (feather and hair) share a similar histological structure. Hair’s dermal papilla cells showed a greater rate of growth as well as a higher and denser cell aggregation when compared to that of feather follicles. In contrast to hair papilla, the feather papilla cells were not able to grow a hair fiber in implanted follicles.
Conclusion: Despite similarities in embryonic development, histological structure and cell growth in vitro, it seems that the signals coming from cultured dermal cells of a mature bird are not able to be understood by hair follicle epidermal cells and can not initiate dermal-epidermal interactions

Abstract Aims: Given the importance of the identification of scientific and commercial communities of Artemia species, especially hybrid populations, some phenotypic and genotypic characteristics of hybrids between A. sinica and A. urmiana were studied in vitro.
Materials and Methods: breeding two species of brine shrimp from hatching to maturity in standard laboratory conditions was provided. Then 64 males and females of each species were separated and mutual hybrid production was done in 50 ml tubes. Daily, the larvae were separated and reared partly and independently. Studies of fatty acids profile and survey by restriction enzyme pattern of 12S-16S mtDNA genome with HpaII enzyme in comparing of the parents and offspring’s of first-generation were done.
Results: Comparing hybrids fatty acids profiles to pure parents showed that some fatty acids amounts are highly dependent on parental origin. In some samples the same restriction enzyme pattern was observed in male parents and the first generation hybrids that was unexpected.
Conclusion: The results revealed that fatty acids profile is strongly influenced by maternal or paternal origin genes of inheritance. Therefore, by directional selection of parents it is possible to produce special Artemia offspring’s with unique phenotypic characteristics. Comparing of enzymatic digestion patterns with the first generated hybrid revealed that the mitochondrial genome (or mitochondria) also is inherited from the sire.

Abstract Aim: The purpose of this study was to evaluate the effects of sub-lethal osmotic stress (325, 350, 375 and 400 mOsm) during cryopreservation in commercially diluter medium (Bioexcell) on Holstein mail cow sperm qualitative characters after freezing-thawing. Material and Methods: Semen was collected from four Holstein mail cows using artificial vagina two times for a week. All of obtained semen mixed together and then were divided into five equal parts. Each part was freeze-melted according to the experimental treatments consisting of different osmotic stresses. 300 mOsm treatment medium was applied as control group. Sperms motility and progressive motility were assessed by computer assessment semen analysis. Also sperms viability, membrane integrity, mitochondria activity and membrane lipid peroxidation of frozen-thawed semen were assessed using Eosin-Nigrosin, hypo osmotic swelling test, Hankok, Rhodamin 123 and TBA procedures, respectively. Results: Results showed that 375 mOsm osmotic stress treatment had the most significant improvement of motility%, progressive motility and viability in comparison with other treatments. Also mitochondria activity in 350 and 375 mOsm osmotic pressures treatments was significantly higher than other experimental treatments (p < 0.05). Different osmotic stresses treatments did not show significant effect on sperm morphology and membrane lipid proxidation. Conclusion: It is seemed that mild and sub-lethal osmotic stresses induction in cow sperm freezing diluters can significantly improved some of sperm qualitative characters such as their motility and viability.

Abstract Aim: In current study capability of differentiation of embryonic carcinoma cells P19 to cardiomyocyte through inducing effects of DMSO and 5-Azacytidin individually or in combinative form in laboratory condition were examined. Material and Methods: In order to differentiate induction of P19, Embryoid Bodies (EBs) formed through hanging drops method during two days. Then EBs have induced by 2 μM of 5-Azacytidin (5-Aza group), 0/5% DMSO  (DMSO group) or both 5-Azacytidin and DMSO (Aza+DM Group) during ten more days. During differentiation beating number per minute counted every two days in all groups by invert Microscopy. Gene expression such as:  α-Myosin Heavy Chain (α-MHC) and Myosin Light Chain (MLC) and Connexin-43 have examined by RT-PCR in last day. To more accuracy, F-actin protein expression also illustrated through immunocytochemistry method. Results: Morphological changes in differentiated cardiomyocytes derived from P19 illustrated in all groups. However the most changes such as significant increasing in size and number of process, their branches and attachment between branches happened in Aza+DM group. Daily beating counter per minute revealed increasing in beating number from 2+2 to 2+8 in all groups. RT-PCR analysis also revealed expression of both α-MHC and MLC in Aza+DM Group. However, F-action has expressed only in Aza+DM group. Conclusion: The Results have shown combination of two 5-Azacytidin and DMSO inducer caused effective cell culture differentiate of P19 to cardiomyocyte.

Abstract Aim: The aim of this study was to assess the protective effect of Meristotropis xanthioides extract on ethanol-induced hepatotoxicity.
Material and methods: The total phenol and flavonoid contents of the species were measured by Folin Ciocalteu and AlCl₃, respectively. Male Wistar rats were divided into three groups: group 1 (control), group 2: received 1ml 50 % ethanol, group 3 : received 1ml ethanol (50 %) plus the extract (500 mg/kg), each group consisted of six rats. All treatments were performed by intragastric administration. Silymarin was used as positive control. Biochemical, histological and morphological analyses were used for evaluation of hepatotoxicity.
Results: The extract had high total phenolic and flavonoid contents. Alcohol consumption significantly increased liver/body weight ratio, amounts of tissue’s malondialdehyde (MDA), H₂O₂ and also liver enzymes in blood in comparison to other groups. Histological examination showed that alcohol consumption injures liver tissue intensely. All these alcohol-induced changes were effectively inhibited by treatment with the extract.
Conclusion: This research confirmed that the species can represent the protective role on ethanol-induced hepatotoxicity and suggested its capacity to apply as a new healthcare food and drug supplement.

Abstract Aim:The aim of this study was to investigate histopathological alterations and their incidence time pattern due to chromium (Cr) exposure during 18 days in gill, digestive gland and foot in freshwater bivalve, Anodonta cygnea.
Material and Methods: 24 bivalve specimens with length range of 12.7-13.3 cm were collected from Semeskandeh region, Sari. They were exposed to 125 µg l^-1 of Cr for 18 days. Bivalves were sampled in days 4, 9 and 18 for obtaining tissue samples of studied organs. Tissue sections of exposed and control group samples were prepared and staining was done using haematoxylin and eosin (H&E) method.
Results: Histology of all of three studied organs of exposed bivalves in comparison with control group showed significant histopathological changes. Hypoplasia and changes in shape and size of gill lamellae, granuloma, hyperplasia and atrophy of haemolymph channels were observed. In digestive gland, atrophy of digestive tubules, loss of digestive and basophilic cells from tubules, and haemocyte aggregation and granuloma in connective tissue were occurred. Hypoplasia of external epithelium, increase of mucus cells, and swelling and tissue rapture in myocyte blocks were observed in foot. Primary signs of histopathological alteration were appeared in fourth day, and in following days appearance of new signs and expansion of affected area was detected in all of investigated organs.
Conclusion: Based on our results, histological changes due to Cr exposure in studied organs in A. cygnea are proposed as appropriate histopathological biomarkers to monitoring Cr levels in aquatic environments and their incidence time pattern can increase fidelity of these indices.

Abstract Aim: The aim of the present study was to stereological evaluate the protective effect of green tea extract on the morphological changes of seminiferous tubules following sodium arsenite exposure in mice.
Material and Methods: 24 adult male NMRI mice were randomly divided into four groups including control, sodium arsenite (5mg/kg/day), green tea extract (100 mg/kg/day) and finally sodium arsenite+green tea extract. Mice were (orally) treated for 34 days. At the end, mice were sacrificed and their right testis were taken out, fixed, processed and stained with heidenhain' azan method. The morphological and changes of testicular tissue was examined using stereological methods. Data were analyzed using one way ANOVA and means difference was considered significant at p < 0.05.
Results: Sodium arsenite treatment caused a significant decrease in the mean seminiferous tubules diameter (p < 0.008), germinal epithelium height (p < 0.001) and the basement membrane thickness (p < 0.011) as well as in the number of spermatocyte (p < 0.003), round spematid (p < 0.028), long spermatid (p < 0.011) and sertoli (p < 0.03) cells compared to the control group. The above changes increased to the control level in sodium arsenite+green tea exteract group.
Conclusions: The results indicate that green tea extract may be useful in reducing the toxic effects induced by sodium arsenite.

Abstract Aim: This study was performed to investigate if silymarin can prevent the adverse effects of sodium arsenite on ram sperm DNA and nuclear integrity.
Material and Methods: Epididymal sperm obtained from Farahani’s ram (Ovis aries) was swim up and divided into five groups: 1. Sperm at 0 hour, 2. sperm at 180 minutes (control), 3. sperm treated with sodium arsenite (10μM) for 180 minutes, 4. sperm treated with silymarin (20 μM) + sodium arsenite (10 μM) for 180 minutes and 5. sperm treated with silymarin (20 μM) for 180 minutes. Ram´s sperm DNA integrity was assessed by SCD (Sperm Chromatine Dispersion) test to study DNA fragmentation and Acridine orange staining was used to estimate DNA denaturation (double-strand DNA versus single-strand DNA). To evaluate sperm nuclear integrity, Diff-quick staining was used and sperm nuclear diameter was measured.
Results: DNA fragmentation percent and nuclear diameter of the spermatozoa were significantly increased and decreased, respectively, in sodium arsenite group compared to the control samples. While this toxicant had no effect on sperm DNA denaturation. In silymarin + sodium arsenite group, silymarin was able to significantly ameliorate the adverse effects of sodium arsenite on these sperm parameters compared to sodium arsenite group.
Conclusion: Silymarin as a potent antioxidant could compensate the adverse effects of sodium arsenite on DNA fragmentation and nuclear diameter of ram sperm.

Abstract Aim: In this study the efficiency of lentiviral gene transfer and cytomegalovirus promoter mediated overexpression has been investigated. Furthermore, the effect of small molecule-mediated inhibition of epigenetic pathways on gene overexpression has been studied. Materials and Methods: Mouse ES cells were transduced with different doses of lentiviruses  harboring Green Fluorescent Protein (GFP) and the percentage of GFP expressing cells was quantified with flowcytometery. The persistence of GFP expression was assessed after 8 days of transduction. Using embryonic stem (ES) cells transduced with a lentiviral vector harboring pancreatic and duodenal 1 (Pdx1) gene, we studied the influence of 5-azacytidin (5-AZA), DZNep, and BIX01294 small molecules on the gene expression system. Results: Lentiviral mediated gene transfer to ES cells with 10 and 20 multiplicities of infection (MOI) resulted in more than 90 percent transgenesis. However, the expression of transgene showed a dramatic decrease during 8 days of post-transduction. Treatment with 5-AZA leads to a 2.5 folds increase in the transgene expression in a permissive culture medium. DZNep also elevated the transgene expression up to 26.5 and 5.9 folds in pluripotency and permissive media respectively. The expression of endogenous Pdx1 gene also increased following DZNep treated. Conclusion: Lentiviral transduction with MOIs more than 10, is an efficient method for transgenesis of mouse ES cells. However, co-application of this method along with the CMV promoter leads to inactivation of the gene expression in long term. DZNep treatment results in reactivation of transgene expression but also can influence the expression of endogenous genes.  

Abstract Aim: The aim of study was designed to determine the created macroscopic abnormalities by Gabapentin during of implantation and organogenesis of fetus.
Material and methods: In this experimental study forty female mice (NMRI type) were used. Female pregnant mice were divided in four groups (each one 10) consist of three experimental groups (I, II, III) and one control (received normal saline). Three experimental groups received 15, 30 and 45 mg/kg/day of Gabapentin respectively by intraperitoneally (ip) injection from day 6.5 (GD_6.5) to day 14.5 (GD_14.5) of pregnancy. Animals were dissected in 18.5 day of pregnancy and their embryos were studied using stereomicroscope from the macroscopical abnormalities. Embryos weight and length were measured and recoded based on CR length. Data were analyzed by T-test, Tukey test and ANOVA using SPSS software.
Results: In all three experimental groups (I, II, III) was observed a significant reduction in comparison with control in body weight and CR length (p < /em><0.001). Also significant increasing was observed in abnormalities appearance such as fetal absorption, organs bleeding and follicular thyroid in comparison with control.
Conclusion: Gabapentin drug is caused fetal absorption, weight and CR length reduction in mice embryo during organogenesis. Also it induces some abnormalities such as hemorrhage and follicular thyroid in embryonic period.

Abstract Aim: The aim of this study was to investigate the effece of mare serum and fetal bovine serum (FBS) on meiotic and fertilization capacity of sheep oocytes in vitro
Material and Methods: Cumulus Oocyte Complexes (COCs) were recovered from ovaries using slicing method. COC’s were washed three time in maturation medium without any serum supplementation. The COC’s were randomly divided into three groups. Group 1 (n = 170) COC’s were fresh control and cultured in maturation medium without serum supplementation. Group 2 (n = 169) COC’s were cultured in maturation medium supplemented with 10% mare serum. Group 3 (n = 167) COC’s were cultured in maturation medium supplemented with 10% FBS. Cumulus expansion was the index of oocyte maturation. To fertilize the mature oocytes, 24 h after maturation oocytes were transferred to the fertilization medium and then sperm was added to the fertilization medium. Twenty four hours post-fertilization, oocytes were denuded mechanically by gentle pipeting, fixed in a mixture of acetic acid and alcohol, stained for 10 min with 1% (w/v) orcein in 45% acetic acid and examined for their fertilization potential.
Results: The rate of oocyte maturation in groups 1, 2 and 3 were 59/82%, 77/55% and 89/27%, respectively. The rate of fertilization in three groups was 19/19%, 39/64% and 50%, respectively. Significant difference was observed between three groups (p < 0.05).
Conclusion: The results showed that the medium contain 10% percent fetal bovine have higher in vitro maturation and fertilization of oocyte rates than mare serum and the control groups

Abstract Aim: The purpose of the present research project was evaluation of the effect of silicon on the severity of peroxidase genes expression and some morphological traits in both resistant and susceptible barley lines under drought stress. Material and Methods: Total soluble protein content, photosynthetic pigments, and total RNA were extracted from the leaves affected by various treatments. Target gene was evaluated by Semi-quantitative RT-PCR analysis using synthesized cDNA. Both the peroxidase enzyme activity by Chance method and proline with Bates method were also evaluated under drought stress treatment. Accordingly, the experiment was analyzed in a factorial test based on the completely randomized design with three treatments of control, drought and silicon-drought (sodium silicate 2 mg / 1 Kg soil) with three replications in a greenhouse. Results: Silicon application caused to increase the amount of total soluble protein and photosynthetic pigments in both lines under drought stress. Semi-quantitative RT-PCR analysis of treatments observed significant differences. Maximum of peroxidase gene expression was observed in the silicon-drought treatment. Antioxidant enzyme activities were at the highest level as shown by gene expression pattern of polyacrylamide gel analysis for treated silicon samples. A further increase was exhibited for proline accumulation caused by silicon application rather than such accumulation in stress treatment compared to control. Conclusion: Silicon reduces oxidative damage induced by reactive oxygen species and with the increase in antioxidant enzyme activities caused to protect plants against environmental stresses.