Cell and Tissue Journal

Abstract Plants produce a big group of secondary metabolites which are used as medicinal compounds. According to recent estimates, global market value of herbal medicines, including medicinal plants and their products, significantly has been increasing. Considering to the fact that most of the world market for medicinal plants, production and supply of secondary metabolites derived from these plants are concerned and the plant secondary metabolites are of high economic value. Chemical synthesis of these metabolites is an expensive process. So production of metabolites by different biotechnological methods such as cell culture is a useful alternative. Molecular recognition and elicitor-plant receptors interaction is a complex process requiring for signal transduction. Biotic elicitors induce secondary metabolites and hypersensitive responses by activation of defense mechanisms. Manipulation of cell culture media by elicitors is an important strategy for inducing secondary metabolism and production of valuable metabolites. Molecular recognition and elicitor-plant receptors interaction is a complex process requiring for signal transduction. Following perception of elicitor signals, rapid defense responses can be organized as follows: increase of ionic currents across the plasma membrane, reactive oxygen species (ROS) production, activation of defense gene expression, structural changes in the cell wall and phytoalexin production. In this study, different aspects of increasing the production of secondary metabolites in cell culture of plants by biotic elicitors is investigated.  

Abstract Aim: In this study, the effect of silence on the expression of the key gene expression of DBOX (which encodes the final enzyme for the synthesis of two alkaloids, Sanguinarin and papaverin) was used by VIGS technique in a species of poppy (Papaver somniferum L.).
Material and methods: A fragment of 350 pairs of alkali from the DBOX gene sequence (within the range of 1112-1462bp) was selected based on the highest number of siRNA production with 21 nucleotides length. After cloning this segment into the pTZ57R/T vector and transferring the vector to pTRV2 viral vector, Agrobacterium inoculation liquid containing silencer was injected into the poppy plants leaves. Primary transgenic plants were selected by PCR reaction using a protein-binding protein coding gene primer (CP) and secondary screening was performed by semi-quantitative PCR technique. In the next step, the samples with the maximum silence (lowest expression) of the gene were examined by real-time RT-PCR technique.
Results: Cloning accuracy in pTZ57R/T and pTRV2 plasmids were confirmed using PCR and enzymatic digestion. Based on the results of semi-quantitative PCR, 5 transgenic plants were selected with the lowest expression for DBOX gene. Based on semi-quantitative PCR results, 5 transgenic plants with the lowest expression were selected for DBOX gene. The results of real-time RT-PCR showed averagely decrease of 81% in the expression of DBOX gene transcriptions in transgenic plants compared to control plants (inoculated with the pTRV2 empty plasmid).
Conclusion: The results generally showed that the VIGS technique could successfully reduce the DBOX gene expression in poppy plants. In addition, the results obtained for this gene can be used to understand the biosynthetic pathway of poppy alkaloids and transgenic plants for metabolic engineering purposes.
 

Abstract Aim: In present study the hepato-protective effects of Tragopogon graminifolius hydro-ethanolic extract (THE) was investigated in an experimental CCl₄- induced hepatotoxicity model.
Material and methods: Six groups (n =6) of male Wistar rats were treated with mixture of 2ml/kg BW of sterilized olive oil and CCl₄ (1:1), along with 200, 400 and 800 mg/kg BW of THE, respectively. Toxicant group received sterilized olive oil 2ml/kg and CCl₄ (1:1) mixture. After 96 hours, liver tissue pathological and serum biochemical parameters (ALT, AST and ALP) studies were done in CCL₄+THE different dosages treated groups in comparison with control.
Results: Results showed that receiving CCL₄ caused acute inflammation and necrosis in rat liver. THE treated groups significantly showed reduction in all of liver tissue biochemical and histological parameters (p<0.05).
Conclusion: It is concluded that T. graminifolius extract has protective chemical compounds such as antioxidants and flavonoids which are able to protect the liver from damages against CCl₄ poisoning by oxidative stresses confrontation.

Abstract Aim: Elaeagnus angustifolia L. is a plant with many therapeutic effects. In this study the effect of aqueous extract of E. angustifolia fruit on Balb/C mouse embryo lens development was investigated.
Material and Methods: Thirty pregnant mice were randomly divided into two equal groups. Control group received drinking water daily and the experimental group received E. angustifolia fruit aqueous extract orally at a dose of 500 mg/kg from zero days to 18th day of pregnancy. After killing all of mice at 18th day gestation, their fetal heads were removed, fixed and processed for histological procedures. Fetal heads were sagitally sectioned at 5 µ thickness and then were stained with hematoxylin-eosin (H&E) and acid fuchsin-light green techniques. Then sections were examined using a light microscope and Motic software.
Results: Results showed that weight and Crown-Rump length mean of fetuses significantly increased in experimental group compared to control. Mean number of fetuses in experimental group was no significant decreased in comparison with control. Means placenta weight and diameter significantly decreased in experimental group compared to control. Also antero-posterior and superior-inferior diameters of lens and lens surface area significantly decreased in experimental group in comparison with control.
Conclusion: These results indicate that using E. angustifolia fruit aqueous extract at 500 mg/kg dose in pregnancy during is caused delaying in mouse embryo lens development.
 

Abstract Aim: The toxic effects of cadmium exposure are mainly due to the production of oxygen free radicals, reduction of cellular antioxidants and oxidative stress, which can lead to cell component destruction, DNA damage, apoptosis and ultimately tissue damage. In this study, the protective effects of N-acetylcysteine, as a substance with antioxidant properties, in cadmium-exposed Wistar mice were investigated by liver histology and measuring the expression of effective genes in apoptosis and cell proliferation.
Material and Methods: Mice weighing approximately 150-200 g were classified into three treatments including, G1) control treatment, G2) cadmium recipient treatment, and G3) concomitant cadmium and N-acetylcysteine treatment and for four weeks received the desired. Then liver tissue samples were taken for histopathological examination and expression of eif4e and mad1 genes.
Results: The results of this study showed that cadmium exposure resulted in serious damage to rat liver tissue, including central artery hyperemia, increased number of inflammatory cells and inflammation in the liver parenchyma, while the use of N-acetylcysteine significantly reduced the mentioned injuries. Also, the use of N-acetylcysteine resulted in a significant reduction in the expression of eif4e and mad1 genes by 2.14 and 2.27 times, compared with mice that received only cadmium.
Conclusion: The results of this study suggest that N-acetylcysteine as an antioxidant can play an important role in preventing tissue damage due to oxidative stress and preventing the induction of cellular apoptosis.

Abstract Aim: Plant growth regulators are able to change the physiological process. Aim of this study is studies of gibberellin (GA3) effects on soluble proteins and root and leaf protein banding profile of short stem Ri T-DNA carrier transgenic Nicotiana tabacum L. plant in comprising with control.
Material and Methods: First, transformation of tobacco plant cv. Wisconsin was confirmed using PCR technique. Then transgenic and non transgenic plants were treated with 0, .2 .4 mg/l GA3 in MS media under in vitro culture for 4 weeks. Soluble proteins as well as SDS PAGE of root and leaves from transgenic and non transgenic plants treated with GA3 were investigated.
Results: Transformation in transgenic plant was confirmed using GUS primers. Assessment of roots and leaves protein of transgenic and non transgenic treated with different concentrations of GA3 revealed that exogenous GA3 had significant impact on increasing of total root protein of root and leaf in both transgenic and non transgenic plants. Also GA3 increased the number of protein bands in electrophoresis pattern.
Conclusion: It is believed that exogenous GA has an impact on root and leaf protein synthesis through transcription and translation processes and also inhibited the activity of protein degradation enzymes.
 

Abstract Aim: Study of gynoecium and female gametophye is important in botany and plant taxonomy. This research was done for studying the developmental stages of ovule in Ranunculus arvensis and comparing with other reported taxa in this family.
Material and methods: Flowers and young buds were collected in different developmental stages, fixed in FAA₇₀ and stored in 70% ethanol. Specimens were sliced using microtome after dehydration and embedding in the paraffin wax and stained. Developmental stages of ovule and embryo sac were studied and photographed using a light monitoring microscope.
Results: Results showed that the ovule is anatropous, unitegumic with 4 or 5 layers and full-nucellus. Embryo sac development is followed monospori and polygonum type. Increasing of antipodal cells and in some cases increasing of synergids were observed. Also their nuclei were polyploid. Archeosporial cells were surrounded by callosic wall after their formation.
Megaspore's tetrads had linear and T-shaped ornamentations. Embryo sac was large with irregular shape and considerably grows longitudinal in its maturation step.
Discussion: Results showed that although the overall pattern of ovule development in the studied species is similar to previous reports, but the tetrad arrangement, increasing of antipodal and synergid cells, and the presence of Hypostase, Postament and Nucellar Cap is the specific characteristic for this species.
 
 

Abstract 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 inductionof breast and prostate­ cancer cell death by extracts, Annexin/PI test were performed. BCL₂ gene expression was analyzed using Real-Time RT-qPCRfor 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.
 

Abstract Aim: In the present study, to compare the toxicity and possible oxidative stress that may result from application of silver nanoparticles and silver ions, the impacts of their different concentrations on some biochemical indices related to protein oxidation in potato (Solanum tuberosum) were investigated.
 Material and methods: potato explants with one node were transferred to MS medium containing 0, 2, 10 and 20 mg.L^-1silver nanoparticles (AgNPs) and silver nitrate (AgNO₃). After four weeks of exposure, in vitro-grown explants were harvested for measurement of various parameters.
Results: Total protein content in explants treated with either AgNPs or Ag ions decreased with increase in silver concentration, except in Ag ion treatment at 2 mg.L^-1, which it increased significantly as compared to control samples. Also, remarkable changes were observed in five bands of protein electrophoresis patterns. The contents of Carbonyl groups were significantly increased relative to control in a dose-dependent manner. In both silver treatments, protein thiols were significantly decreased, while non-protein thiols were amplified. Moreover, explants treated with Ag ions showed higher protease activity and total antioxidant power as compared to AgNPs treated ones.
Conclusion: Based on the results, it could be concluded that oxidative damage to explants treated with AgNPs was much more than explants under a similar mass of Ag ions. In addition to the release of silver ions, special effects associated with nanoparticles could be involved in the oxidative stress induced by AgNPs in potato explants.

Abstract -
Aim: The purpose of this research was to design and prepare a suitable gene construct and transfer it to the tobacco plant and analysis of transgenic plants.
Material and methods: Seed-specific construct that containing Napin promoter, Ω sequence, GUS gene, and SAR sequence was prepared in pBI121 plasmid and proliferated in E.coli.. Then tobacco leaf explants were inoculated with LBA4404 agrobacterium strain by standard protocol. Selection of regenerated shoots also were performed in a selection media (co-culture medias + 25 mg/L Kan + 200 mg/L Cef). Transgenic plants were analyzed by PCR, RT-PCR and histochemical assay.
Results: Analysis of regenerated plantlets by using PCR and specific primers of nptII and GUS indicated that transfer of these genes to plantlets was successful. RT-PCR reaction results showed that nptII is transcribed in both tissues while GUS is transcribed only in the seed tissue. This finding was expected because Nos promoter (which controls the nptII transcription) is a constitutive and Napin is a seed specific promoter.. Expression and activity of Beta-glucuronidase enzyme in seeds of selected plants was confirmed by SDS-PAGE and histochemical assay.
Conclusion: The result of this research showed that designed gene construct was appropriate,  because Napin promotes the expression of GUS gene in the seeds and Omega sequences have also been effective in increasing of transgene expression.  In the fallowing, this construct can be used for the production of recombinant proteins by replacing valuable genes with GUS..
 

Abstract Aim: The aim of this study was to evaluate the effect of Valproic acid (VPA) as a histone deacetylase inhibitor on decrease of microglia /macrophage activaty and nervous tissue destruction after spinal cord injury of rat (SIC). Material and Methods: To SCI, contusion model was used. Ten contused rats were equally divided into two groups. Control group did not receive any injection and treatment group received valproic acid (400 mg/kg) intraperitoneally daily for two weeks. Rats were killed at 28 days post injury then damaged spinal cord was removed and examined for H4 acetylation, ED-1 and OX-42 positive cells using immunohistochemistry procedure. Also the cavity volume percentage in 4200 μm length of the spinal cord (central, rostral and caudal regions from the injury epicenter) was assessed for each sample. Results: Results showed increases of histone H4 acetylation and decreases of ED-1 (lysosomal marker) and OX-42 (microglia marker) positive cells. Also the percentage of cavity volume in valproic acid-treated group compared to control (untreated) showed significant decreases. Conclusion: Prescription of valproic acid in the early stages of SIC decreases microglia/macrophages activity, and neural tissue damage in a spinal cord injury model.

Abstract Aim: In this study the effects of stress on induction of structural chromosomal abnormalities on L929 cell line using micronucleus assay on cytokinesis-blocked binucleated cells were investigated in vitro. Material and methods: L929 cells were cultured in DMEM containing 10% FBS. The cells were divided into four groups including; control, 2Gy gamma-irradiated cells, and cells treated with doses of 25, 50 and 100 µg/ml hydrocortisone, and cells co-treated with these three doses of hydrocortisone and irradiation. The treated cells were harvested, stained and chromosome abnormalities were scored using micronucleus assay. Results: Results showed that hydrocortisone did not induce micronuclei as compared to the control group. However, the frequency of micronuclei in cells co-treated with doses of hydrocortisone and irradiation was significantly higher than cells treated only with gamma irradiation (p < 0.05). Conclusion: According to the data of this study, stress hormones are not able to induce any chromosomal abnormalities; however, they are able to increase the cell susceptibility to clastogenic effects of irradiation. Key words: Stress, Hydrocortisone, L929, Gamma ray, Micronucleus      

Abstract Aim: In this study we were investigated the importance of the preparation method of the substrate based on PLGA polymer by electrospinning and freeze-drying method. To investigate the effect of nanotopogeraphy on cell behavior, the prepared nanofibers were compared with each other in three speeds of the collector.
Material & Methods: PLGA substratums were made by electrospinning and freeze-dying. The morphology of the structures was compared by use of the Scanning electron microscope images. Mouse fibroblast cells (L929 cell line) were seeded on substrates to determine the cell viability and followed by MTT assay.
Results: Scanning electron microscope images showed that, by increasing the speed of collector, the nanofibrous orientation increased. MTT assay (p < 0.05) showed in 24 h that PLGA substrates prepared by freeze-drying method were provided the appropriate attachment for the cell by producing a porous structure. Cell viability was significantly increased on the PLGA nanofibers with increase of fibers regularity after 48 and 72 h cell culture. This feature did not change on freeze-drying substrate.
Conclusion: The findings were indicated that electrospun PLGA nanofibers had the better performance to provide cell behavior. It was expected because of similarity to the structure of the natural extracellular matrix. It seems that aligned fibers acted as a positive factor to support cell proliferation.

Abstract Aim: Para-Nonylphenol is an environmental pollutant which can generate free radicals causing tissue damage. The aim of this study was to investigate the effect of vitamin E as a strong antioxidant on kidney tissue in rats treated with para-Nonylphenolusing’stereological technique.
Material and Methods: Twenty fourmaleWistar rats with the mean body weight of 198±20 g were randomly divided into 4 groups (n=6), control, vitamin E (100mg/kg/day), Para-Nonylphenol (250mg/kg/day) and Para-Nonylphenol+ vitamin E, and treated orally for 56 days. Finally, the rats were weighted and their right kidneys were taken out, fixed, sectioned, processed and stained using Heidenhain̕& azan method. The kidney tissue was then studied using stereological methods. Data were analyzed using one way ANOVA and means difference was considered significant at p < 0.05.
Results: The mean kidney weight, the total volume of kidney, the volume of cortex and medulla, the volume of proximal and distal tubule lumen, interstitial tissue and the volume of glomerulus and tuft significantly increased in Para-Nonylphenol group compared to the control ones, while the volume of proximal and distal tubules epithelium and the volume of Bowman’s space reduced significantly.
Conclusion: Vitamin E could compensate many adverse effects of Para-Nonylphenol on the kidney tissue. Consequently, vitamin E could be used as a potential antioxidant (supplement) against NP intoxication.
 

Abstract Aim: In this study, the effects of recombinant human growth hormone and Gemcitabine alone and in combination with each other were investigated on the lung fibroblast cell lines.
Material and methods: Human growth hormone at 10-400 ng/ml and Gemcitabine at 1-100 μg/ml concentrations were treated with the human lung fibroblast cells and the analysis were performed with MTT assay, propidium iodide staining and scratch assay.
Results: Studies of the cell cycle and MTT assay revealed the effect of growth hormone on the progression of the cell cycle and exit of G1 phase and the cell proliferation, and the inhibitory effects of the cell cycle and growth process by Gemcitabin compared with the control sample. The results of the scratch test indicated that the growth hormone at its effective concentrations caused increased the cell migration compared to control, while the Gemcitabin reduced the migration of the cell to the control.
Conclusions: Using growth hormone in combination to gemcitabin during chemotherapy treatment, it seems that we can reduce the rate of the neem normal cells death. Since these cells are more sensitive to growth hormone´s effect than cancer cells.
 

Abstract Aim: In this study, the effect of newborn rat brain extract to induce neuronal differentiation in P19 embryonal carcinoma (EC) stem cells was investigated.
Material and Methods: Newborn rat brain extract was collected in sterile condition and the concentration of its total protein was determined. Then, the amount of cell viability was determined after treatment with brain extract. In order to differentiation, the embryoid bodies resulted from cells suspension culture were exposed to culture medium containing 3% serum that supplemented by the extract, for 7-14 days. Specific staining and real-time PCR methods were applied to evaluate neural differentiation.
Results: Cresyl violet staining confirmed neuronal morphology of the differentiated cells. The gene expression of neural specific was confirmed by real–time PCR. Developing brain extract, which contains numerous neorotrophic factors, could induce expression of synaptophysin (presynaptic membrane protein) and nestin (intermediate filament of stem cells in the neural tube) genes. In addition, the expression of transcription factor Nanog, an important factor for pluripotency and self-renewal of stem cells, decreased under the influence of newborn rat brain extract.
Conclusion: The results of the present study indicated that newborn rat brain extract can induce neuronal phenotype as well as neuronal specific gene expression in P19 stem cells. This study suggests the potential use of combined newborn rat brain extract and stem cell therapy to improvement of deficits in neurodegenerative diseases.

Abstract Aim: Cisplatin is one of the most successful drugs to fight many cancers, but its nephrotoxicity limits its administration. The aim of this study is investigation of the effect of human mesenchymal stem cells secretory factors on Cisplatin-induced acute renal failure in rats.
Material and Methods: In this study, male wistar rats were used. Animals were randomly divided into four groups and six animals were in each group. One group was considered as control which this group received no injections. Other three groups were treated with intraperitoneal 5 mg/kg Cisplatin once at the beginning of the experiment. Then one of these three groups was injected with conditioned medium of human mesenchymal stem cells and other groups were injected with medium without secretory factors intraperitoneally for three consecutive days. After five days Cisplatin injection, kidney and blood samples from all of examined animals were collected for histological and biochemical studies.
Result: Results showed significant decrease in the kidney injury level, blood serum BUN and creatinine amount. Also
Also body weight increasing significantly was observed in conditioned medium of human mesenchymal stem cells receiver group in comparison with treated Cisplatin and non secretory factors receiver groups.
Conclusion: Results of this research suggest that human mesenchymal stem cells secretory factors can be affect against Cisplatin-induced nephrotoxicity.

Abstract Aim: In accordance with conflicting results about the effect of dietary supplements on exercise-induced inflammatory responses, this study was conducted to identify the effect of 14-day caffeine supplementation on the response of serum C-reactive protein (CRP) and peripheral blood leucocytes by following one bout downhill running in male non-athletes.
Material and Methods: Eighteen male volunteer non-athletes (aged 25±3 years, body fat 13±2 and VO2max 50±4 ml/kg/min) in a semi-experimental, randomized and double-blind process were allocated equally into supplement and placebo groups.
Each subject received caffeine or dextrose (5 mg/kg body weight/day) for 14 consecutive days.
After the supplementation (14 days), all subjects were participated in one bout downhill running on a treadmill (-15% incline) for 30 minutes with 65% VO2max.
Changes in serum CRP and peripheral blood leukocytes were counted and determined in four phases (the base line, after the supplement period, immediately and 24 hours after the exercise).The normal data were analyzed by repeated measure ANOVA, Bonferroni and independent t test at α≤0.05.
Results: The results show that the caffeine supplementation has no significant effect (P>0.05) on the basal inflammatory indices. The serum CRP and peripheral blood leukocytes were counted and significantly increased after following the exercise protocol and higher than the baseline levels until 24 hours later (p < 0.05). However, the change of serum CRP and peripheral blood leukocyte counts in placebo group was significantly higher than in caffeine group (p < 0.05).
Conclusion: Based on the findings we can conclude that 14-day caffeine supplementation can probably decrease exercise-induced inflammatory response (CRP elevation and Leukocytosis) following 30 min downhill running in male non-athletes.
 

Abstract Aim: Mangifera indica is an evergreen tree belongs to the Anacardiaceae family, in Iran it is found in the Hormozgan and Baluchistan provinces. Till now, no research has been carried out regarding the structural characteristic and development of this species; therefore in the present study structure of vegetative organs and developmental characteristics of reproductive stages was investigated.
Material and Methods: The vegetative organs were cut and fixed in glycerin and ethanol. The young flowers and buds were sampled at different developmental stages. Samples were fixed in FAA and sliced using a microtome. Staining was carried out with Eosin and Haematoxylin.
Results: The primary and secondary structures of root, shoot, petiole, leaf, inflorescence axil and shoot apical meristem showed similarity to dicotyledons. Generative organs contained androecium with 5 stamens which one was reproductive while others were sterile. The anther had 2 parts and 4 pollen sacs. The tapetum layer was secretary type and pollens were three porated. The gynocecium had a carpel with one cameotropous ovule and one tegument. The placentation was of a basal type. The secretary tissues were seen in vegetative organs of the plant.
Conclusion: Base on the anatomical findings, we observed a characteristics similarity between this plants and other Angiosperms. With respect to the results of the developmental stages of reproductive organs new information for recognition of this plant was obtained.

Abstract Aim: In the current study, CM anticancer function of the MCF7 cells was investigated in vitro.
Material and Methods: Adipose stem cells were extracted from the cesarean women’s abdominal fat with their written informed consent at the Velayat hospital, Damghan. CM was prepared for fourth passage of hASCs that was cultured in serum-free medium for 72h. MCF7 cells were exposed to CM for 24 and 48 hours. Then cell proliferation rate, survival and apoptotic gene expression were determined using MTT, cell counting (hemocytometer) and RT-PCR.
Results: CM-treated MCF7 for 24 and 48h, showed a significant decrement in cell proliferation and viability as compared to the cells cultured in medium containing serum (control). In addition, caspase3 gene expression of CM-induced cells was increased significantly at 24 and 48h as compared to the control group. While, CM-treated cells showed a significant increment of caspase9 gene expression after 48h induction.
Conclusion: It was concluded that CM induced apoptosis by activation of caspases and reduced proliferation rate and survival of MCF7 cancer cells. Therefore, it seems that conditioned medium can be used as a supplement with other anti-cancer therapies.

Abstract Aim: Main goal of this investigation were verification of gene's behavior which located on RNO15 with allelic imbalance during Endometrium cancer development.
Material and methods: BDII/Han females were crossed to males from two other inbred rat strains known to have low incidence of EAC (BN/Han and SPRD-Cu3/Han). To generate F2 rats the F1 animals were intercrossed. In addition, backcross populations were generated by crossing male F1 rats to female BDII rats.  In order to characterize these tumor-specific genetic aberrations in greater detail, we applied molecular cytogenetic methods (FISH, chromosome painting) to 18 rat endometrial adenocarcinoma (EAC) cell cultures.
Results: Analysis in the tumors by FISH analysis showed the presence of small regions of amplifications.
Conclusion: These changes were cryptic so that they occurred inside cytogenetically normal-looking chromosomes. We detected rather specific chromosome aberrations (translocations, deletions, amplifications) often leading to copy number changes in small DNA segments (gains, losses).
 

Abstract Aim: The aim of this study was to  investigate the effect of different concentrations of Aluminum oxide nano-particles on seed germination, root length, and the amounts of photosynthetic pigments, total protein content, and changes in activity of some antioxidant enzymes and carbohydrates content in Phaseolus vulgaris. Material and methods: Experiment was performed under greenhouse conditions and completely randomized designed with four replications. Plants were exposed to different concentrations (0.01, 0.5 and 1 g/L) of nano-Aluminum oxide and the physiological and biochemical characteristics of treated plants were compared with control ones. Results: The results showed that treatment by Aluminium oxide nano-particles had a positive impact on the seed germination, root length, total chlorophyll and chlorophyll b content and also sugar content. Decrease in the content of chlorophyll a, protein and catalase activity was observed in the treated plants in comparison to control. Aluminium oxide nano-particle didn’t have a significant effect on seed germination speed and peroxidase activity. Conclusion: Based on the results of this study, nano-particles are able to have a positive effect on some developmental and physiological characteristics of Phaseolus vulgaris L. This indicated that applied nano-particles are not toxic in the used concentrations. Decreasing in characteristics (chlorophyll a, total protein and catalase activity) was observed that could be related to plant resistance.

Abstract Aim: The aim of this research is the investigation of anti- inflammatory effect of silymarin on the treatment of colon ulcer induced acetic acid in mice Balb/C spicies.
Materials and methods: In this study, 32 mice are divided into 4 groups (n=8).These groups include: control group with no colitis report, the Sham group with colitis report that are not treated and the groups with colitis report that received 40 and 80 mg/kg B.W. of silymarin orally.The groups with silymarin treatment initially received silymarin for 14 days before induction of the disease and then for 1 week after effected by disease. The drugs are fed to the mice per oral. To induce colitis, 1 ml of acetic acid (%4) is injected directly into the rectum. In the one week after induction of colitis, the animals sacrificed and the damages of the colon investigated with Murthy method. Also, the concentration of alkaline phosphatase enzyme measured. Also, tissue oedema  verified.
Results: The results shown the concentration of alkaline phosphatase enzyme has been increased significantly (p < 0.05) in colitis group in comparision with control group whear as the concentration of these enzyme has been decreased significantly (p < 0.05) in treatment groups in comparision with colitis group. Also, acetic acid causes severe inflammation and damages of the colon.
Conclusion: It is concluded treatment utilized silymarin can be considered as a suggestive way.
 
 

Abstract Aim: The aim was to study the histology of somatic embryogenesis stages in Dorema ammoniacum D. using tissue cultures as an efficient method for propagation of the plants with restriction in cultivation. Material and methods: Various explants were cultured on MS (Murashige and Skoog ) medium containing  30 g l^-1 sucrose and 7 g l^-1 agar as well as supplemented with different concentrations of  auxin and cytokinin. After eight weeks, different stages of development of somatic embryos in embryogenic callus were evaluated using histological method. Results: Out of different explants, only zygotic embryo was able to induce embryogenic callus. Presence of NAA (α-Naphthaleneacetic acid) increased callus induction and somatic embryogenesis. The highest callus formation was observed in MS medium supplemented with 1 mg/L NAA and 0.5 mg/L BA (N6-Benzyladenine). Different stages of somatic embryos including: globular, heart-shaped, torpedo and cotyledonus stages were observed in the prepared sections of callus. Conclusion: Using zygotic embryo the in vitro somatic embryogenesis of Dorema ammoniacum D. was possible. In addition the histological examinations of embryogenic callus showed the existence of the different stages of embryonic development.  

Abstract Aim: The present study was designed to investigate the effects of chitosan/poly vinyl alcohol scaffold incorporated by NRBE (neonatal rat brain extract) on neuronal differentiation of P19 EC (embryonal carcinoma) stem cells.
Material and Methods: In order to induce neuronal phenotype, P19-derived embryonic carcinoma stem cells were cultured on a rat chitosan/poly (vinyl alcohol) scaffold with newborn rat brain extracts. To evaluate the survival potential, migration and differentiation of the three-dimensional culture cells, these cells were grafted to the developing central nervous system of the chick embryo. Finally, the transplanted cells were detected using specific staining and immunofluorescence methods.
Results: Cresyl-Violet specific staining confirmed the neuronal phenotype of the transplantation cells. Also, the expression of specific neural protein, synaptophysin, was shown using the immunofluorescence process.
Conclusion: The results of this study showed that P19 embryonic carcinoma stem cells can be used as a source of pluripotent cells in transplantation studies in order to investigate cellular and molecular aspects of developmental and cellular differentiation.