Cell and Tissue Journal

Abstract Aim: In this study, with the aim of optimizing hairy root growth conditions, leaf, hypocotyl and root explants were inoculated by strain ‘A13’ of Agrobacterium rhizogenes. Then effect of four basal MS liquid media was considered with different combinations and temperature on rate of hairy roots growth.
Material and methods: In order to hairy root induction via spary and leaf disc methods, two strains of Agrobacterium rhizogenes (A13 and 9534) were used. For confirmation of transformation but not infection of hairy roots, PCR reaction was performed through specific amplification of rolB and virD genes. Obtained hairy roots in a basal liquid MS medium were cultured under different treatments in based on completely randomized design (CRD) with three replicates and dry weight of hairy roots was measured after two months.
Results: Hairy roots were observed in root, hypocotyl and leaf explants after 8 days. The highest percentage of transformation was found in leaf explants. The obtained results showed that the highest (0.19 gr) and the lowest(0.05 gr) rate of hairy root dry weight related to basal liquid MS medium containing 30 g/l sucrose in 35°C  and basal MS liquid medium containing 0.2 gr _∕ l NAA hormone,respectively,.
Conclusion: The results indicate the main role of culture medium compounds and temperature in enhancing of hairy root biomass.

Abstract Aim: In this study, it was done to evaluate the efficiency of both PRP and Fibrin Glue scaffolds in producing suitable environment for the growth of mesenchymal stem cells.
Material and Methods: In this study, the preparation of PRP and Fibrin Glue Scaffold were carry out and Mesenchymal Stem Cells (MSCs) isolated from adipose tissue. The mesenchymal phenotype of these cells was determined by mesenchymal surface marker using flow cytometry. Then, MSCs were cultured, and on the third passage (P3)stage, they were seeded separately on the two scaffolds and after 48 hours of cell culture, the ability of the scaffolds seeded cells was evaluated by MTT assay for cells viability.
Results:Flow cytometry results showed that human adipose-derived mesenchymal stem cell expressed CD44, CD90 and CD105 surface markers. Also, the results of this study showed that the active PRP could be creating a more suitable environment for the survival and proliferation of mesenchymal stem cells in compare with Fibrin glue
Conclusion:It is suggested that the PRP as a protective scaffold could be used for mesenchymal stem cells growth to provide effective strategies in order to tissue engineering development and regenerative medicine.

Abstract Aim: The purpose of this study is to investigate the pro-apoptotic and anti-metastatic potentials of Lippia citriodora alcoholic leaf extract on A2780 ovarian cancer cells and to evaluate the expression of E-cadherin as one of the most important marker in metastasis.
Material and Methods: A2780 ovarian cancer cell line was prepared from Pasteur cell bank and cultured in RPMI1640 medium containing 10% FBS and 1% antibiotic. After cell seeding and treatment with different concentrations of extract (10-400 µg/ml), the cells viability and pro-apoptotic potential were examined by MTT assay and acridine orange/ propodium iodide, respectively. Caspase-3 assay and anti-invasive effect were analyzed by migration assay and assessment of E-cadherin expression, respectively. Then, one way ANOVA test was employed for analysis of quantitative data.
Results: Data indicated that L. citriodora alcoholic extract attenuated ovarian cancer cell viability. Acridine orange/ propodium iodide and caspase-3 assays showed that this extract in IC50 concentration (100 µg/ml) induced apoptosis mainly through caspase dependent pathway in the ovarian cancer cells. Migration assay and RT-PCR exhibited that this extract has anti-invasive capability and by up- regulation of E-cadherin prevents the loss of attachment between cancer cells.
Conclusion: The results revealed that L. citriodora alcoholic extract has apoptosis inducing capacity and ability of restoration E-cadherin expression in A2780 cancer cells, which it able to suppress invasive potential of ovarian cancer cells.
 

Abstract Aim: To investigate spermatogenesis in the Marsh frog living in Makhmalkooh, Khorramabad, and determine which type of spermatogenesis found in this species.
Material and Methods: 34 male frogs were collected from the study area during September to February. First, the specimens were identified by measuring biometric, meristic characters and according the present keys. Then, abdominal cavity was dissected and the gonads were out and processed to prepare for histological sections methods. The tissue sections were finally examined for histological features and three quantitative parameters.
Results: Microscopic observations showed that in this species the event of sperm releasing starts in March and continue to June, during which spermatogenic cysts are observed on the wall of seminiferous tubules. This event ceases during July to September when spermatogenesis enters to its resting phase. During the resting phase the cysts are not found in the seminiferous tubules. A regenerative phase was distinguished during September to March as the number of cell increased in germinal layer. Quantitative surveys also illustrated that during March to September, diameter of the tubules and their germinal layer initially increased and then declined while the lumen’s diameter showed an opposite flux when compared with two other parameters.
Conclusion: Based on the results presented here, it seems that in this species the spermatogenesis does not occur as continuous but it is carried out in the form of semi-continuous or potentially continuous.
 

Abstract Aim: The aim of this study is determining the sensitivity and specificity of simultaneous use of both DNA methylation of RASSF1A gene and expression of FHL1 gene in the differential diagnosis of benign tumors from malign papillary carcinoma tumor in the thyroid gland.
Material and Methods: 160 samples of patients with malign thyroid tumors (80 samples) and benign thyroid (80 samples) were used in this study. The RNA was extracted from paraffinic samples, and then cDNA was made from it. Gene expression was investigated by Real Time PCR mixture containing the Cyber Green fluorescence material. After DNA extraction, the hypermethylation of the gene was performed using COBRA method. Finally, an epidemiological calculation was used for calculating the effects of sensitivity and specificity of two simultaneous tests.
Results: There was a negative significant correlation (P ≤0.05) between promoter hypermethylation of the RASSF1A gene and the FHL1 gene expression. Net sensitivity of simultaneous use of the promoter methylation of the RASSF1A gene (quantitative and quantitative) and expression of the FHL1 gene were 98.2% and 79.32%, respectively. The Net specificity of simultaneous use of the promoter methylation of the RASSF1A gene (quantitative and quantitative) and the FHL1 gene expression test were 39.47 and 75.29%, respectively.
Conclusion: simultaneous use of the hypermethylation of the RASSF1A gene promoter as a qualitative and expression test for FHL1 gene is more sensitive and more reliable in differential diagnosis of benign tumors from malign papillary carcinoma tumors.
 

Abstract Aim: The main goal of this research was to prepare a three-dimensional matrix from gingival palate tissues and investigate the possible application of this scaffold ​​in cell culture and tissue engineering.
Materials and methods: In order to fabricate the scaffolds, the biopsy samples of human palate gingival tissue were preparated surgically and divided in 5 groups then decellulization of the samples were carried out via physical method (put in nitrogen tanks and rinsing with distilled water) as well as chemical method using different concentrations of SDS (0.1%, 0.25%, 0.5%, 0.75% and 1%). Furthermore to evaluate the scaffold prepared with 1% SDS, embryonic like cells from blastema tissue were seeded on the three-dimensional scaffold.
Results: Concentration of SDS below 0.5% caused significant reduction (p < 0.05) of decellulization of the tissues. Microscopic studies of blastema tissue on the scaffold in different days revealed the penetration, migration, adhesion and differentiation of the cells.
Conclusion: This study showed that, it is possible to prepare a natural scaffold frome palatal gingiva tissue using SDS treatment. On the other hand, the results of histologic studies showed that the decellulized scaffolds of palatal gingival might be suitable as three-dimensional bioscaffold for movement, adhesion, differentiation and migration of cells. More investigation is needed to determine the identity of the differentiated cells which further it can help to improve our knowledge about cell-matrix interaction.
 

Abstract Aim: Researchers have made considerable efforts to determine the significance of particular ABO antigens to diseases susceptibility such as cancer and infections. O blood group people have a higher risk than other blood groups for catching cholera. Thus relationship between ABO blood groups and catching disease is unknown remained. The aim of this study is to determine the lymphocyte phenotypic profile in ABO blood groups.
Material and Methods: Peripheral blood samples collected from forty health male people with different ABO blood groups (each group=10). All of studied people were in 18-25 years old ranges and had a closely similar genetic background. The samples were studied using a FACSort flow cytometr for determination of CD3+T-lymphocytes phenotype and their subsets CD4+, CD8+ and Treg cells (CD4+/CD25+/Foxp3+), CD19+ B-lymphocytes and their subsets CD5+ and CD5‾ B cells.
Results: There were no significant differences in the frequency of examined lymphocyte subpopulations (CD8+ and Treg lymphocytes, and B- lymphocyte subsets) between the ABO blood groups. But, the CD4+ T lymphocytes percentage in B blood group was higher than other groups (p < /em>=0.05).
Conclusion: High frequency of CD4+T lymphocytes in B blood group may be reduced catching some disease such as bacterial and parasitic infections.
 

Abstract Aim: In this study, the histology effect of caffeine administration was evaluated on retina and PAX6 expression in rat neonates.
Material and Methods: A total of 24 pregnant rats were divided into three groups. Intraperitoneal injection of saline in the control group and also 50 mg/kg and 100 mg/kg caffeine injection from the ninth day of gestation was done. At 20 or 21 embryonic day, neonates on the first and fifth days were euthanized anesthetized and the eyes were isolated and fixed. The neonates were examined for abnormalities. The Retina Histological and morphometric evaluations, eye tissue fluorescence changes, and PAX6 gene expression were investigated by Real time PCR.
Results: The results showed that the effects of maternal caffeine treatment on the retina in neonates are associated with atrophic changes and cell counts of ganglion cell layer, retinal thickness and thickness of the inner plexiform layer, in the animals were treated with caffeine, were significantly reduced (p < 0.05). Also caffeine induced behavioral changes in mothers and tissue fluorescence and significantly increased (P <0.05) the PAX6 gene expression in the groups receiving caffeine.
Conclusion: Administration of caffeine during pregnancy can induce histopathological changes in organogenesis and developing in rat neonate’s retina.
 

Abstract Aim: In this study, the effects of simulated microgravity on the apoptosis and expression of p75NTR gene were investigated in the adipose derived stem cells before and after the neural differentiation.
Material and Methods: Human adipose derived stem cells were isolated, cultured and differentiated.  A single-axis clinostat apparatus was used to simulate microgravity for 6, 24 and 72 hours. Real time PCR technique was used for gene expression analysis after extraction of RNA of samples. Cell viability was assessed by MTT assay and apoptosis rate was calculated by Annexin V staining.
Results: Our results showed that microgravity led to a significant decrease in p75NTR gene expression in adipose derived stem cells. However, microgravity had no significant effect on viability of cells before and after differentiation, but apoptosis in undifferentiated cells was decreased in contrast to controls.
Conclusion: Due to reduction of apoptosis and increment of differentiation potential of cells, microgravity can be introduced as a powerful tool and also new condition for cell culture and neural differentiation for achievement to effective cell therapy. 
 

Abstract Aim: Plant production from tissue culture is of high importance from genetic engineering point of view, so in this study, optimization of tissue culture conditions for callus production and plant regeneration of Salsola arbuscula were examined. Material and Methods: Seeds were planted MS medium and after two months, roots, stems (internodes) and leaves explants were transferred to the MS medium with different concentration of 2,4-D and Kinetin hormones for production callus. Plant regeneration was also examined on the various media. Results: Results showed that the best medium to induce callus production were medium of MS + 2,4-D (1 mg/L) + Kin(1 mg/L) and the best explants were roots. So, direct shoot regeneration produced in the medium MS + Kin(1 mg/l) and MS + Kin (0.5 mg/L). Conclusion: Despite production of callus on medium containing only 2,4-D, combined two hormones, auxin and cytokinin, increased the rate of callus production. Also direct shoot regeneration has occurred in medium without auxin. So, production rate of callus and plant regeneration are dependent on amount of external plant growth regulators, and also, amount of external plant growth regulators significantly dependent to genotype and amount of internal plant hormones.

Abstract Aim: The purpose of this study is to examine DJ-1effect on increasing survival of dopaminergic (DAergic) cells against parkinsonian toxicity.
Material and Methods: First recombinant lenti-viruses transporters were produced with both DJ-1 and reporter Jred genes and were used to cells infection. To this end, three lentiviruses vectors namely transporter, packaging and envelope were applied for co-transfection of HEK-293T cells as virus-producing cell line. 24 and 48 hours transfected cell media were collected and concentrated till lentivirus stock generation. Following transduction of DAergic PC12 cells with this concentrated virus stock, the infected cells overexpressed DJ-1. After treatment of these transduced cells with the 6-OHDA toxin, their survival rate was measured in comparison with control.
Results: Transfection HEK-293T cells steps and PC12 cells transduction with the virus stock were done successful, because reporter Jred gene expression was observed using fluorescence microscope in both steps. Then DJ-1 overexpression was proofed using RT-PCR method. Next experiments indicated that DJ-1 overexpression causes significant increase in PC12 cells survival against produced toxicity of 6-OHDA. PC12 cell survival percentage that had DJ-1-overexpressing was 30% more than control cells survival percentage and this increasing was statistically significant.
Conclusion: increasing of DJ-1 expression in DAergic PC12 cells significantly increased their resistance and perpetuity against 6-OHDA neurotoxicity.

Abstract Aim: The aim of this research was cloning and expression of human Midkine coding gene (mdk) in Escherichia coli that achieved in a laboratory-scale experiment.
Material and Methods: Methods were included cell culture, RNA extraction, cDNA synthesis, cloning techniques, induction of expression by IPTG (isopropyl thiogalactosidase), expression evaluation using polyacrylamide gel and confirmation by Western blot techniques.
Results: Midkine gene was cloned in pET-21a (+) and then transformed into Origami strain of E. coli. This growth factor was expressed in cytoplasmic level by a colony containing pETmdk recombinant after 16 hours incubation at 18°C and 250 rpm mixing. Expression of histidine tagged 13 kD protein confirmed by Western blotting technique.
Conclusion: Because Origami strain is trxB and gor genes mutant strain its cytoplasm is an oxidizing environment. Due to this, it enhances disulfide bond forming, therefore it seems that after expression of midkine, cysteine residues make an intra-molecular disulfide bridge and remains in soluble form. These conditions provide suitable environment for the proper folding of the protein and consequently solubilization of the protein.

Abstract Aim: This study aimed to produce recombinant hybrid protein, DT389GCSF, in E. coli BL-21(DE3) in a soluble and active form and to purify it and evaluate its cytotoxicity.
Material and Methods: The protein, His6-tagged DT389GCSF was expressed in E. coli BL-21(DE3) in a soluble form at 28 °C. Then it was purified by affinity chromatography on nickel sepharose column. Finally, the function of purified recombinant protein was evaluated by MTT assay on HL-60 cancer cell line.
Results: The yield of expression and purification of DT389GCSF were determined at about 12 and 80 percent, respectively, using Image J software. The IC50 value upon 48 hours of exposure of DT389GCSF toward cell line was about 0.00017±0.000019 M.
Conclusion: By reducing the temperature and using the intracellular mechanism, the refolding of hybrid protein, DT389GCSF, can be observed in a proper and active form. As a result, in vitro production of relevant immunotoxins, by using this method, the steps of inclusion body production can be neglected.
 

Abstract Aim: The aim of this study was to collect and identify Fusarium of Elegans and Martiella-Ventricosum sections related to cucurbits plant in Tehran province and evaluation the efficiency of ITS-RDNA RFLP polymorphisms as a molecular marker in assessment of relationships between these Fusaria. Materials and Methods: During the cropping season, sampling from cucurbit field was done from the root, crown, stem up to height of 15 cm, as well as the soil (rhizosphere). Fungal isolates were cultured on PPA culture medium, purified by single sporulation method, and then identified based on morphological characteristics. Genomic DNA of fungi was extracted. The genomic DNA of Fusarium isolates was extracted and ITS-rDNA region was amplified with ITS1 and ITS4 primers. PCR products were digested with SmaI, BglΠ and MboI restriction enzymes and then loaded on 2.5% agarose gel. Data analyses were performed by NTSYS-PC V2.02 software. At first, a data matrix of the presence (1) or absence (0) of each band was drawn for each isolate. Then using the SM similarity coefficient, the genetic distance matrix was produced and the similarity dendogram of isolates was drawn based on the SAHN coefficient and the UPGMA method. Results: A total of 95 Fusarium isolates belonging to species of Fusarium solani and Fusarium oxysporum were identified. Of which, 45 isolates belonging to F. solani and 50 isolates were identified as F. oxysporum. In this research, based on morphological characteristics, F. redolens isolates were not differentiated from F. oxysporum isolates and both were placed under F. oxysporum group. PCR reproduction of ITS region result in fragments in size of 550±25 bp in F. oxysporum isolates, and 575±25 bp in F. solani isolates. Bgl Π enzyme had no cleavage site in ITS products in both species, neither in F. solani nor in F. oxysporum. SmaI enzyme had one cleavage site in F. solani isolates and produced two fragments (350bp and 230bp) but had no cleavage site in F. oxysporum isolates. The MboI enzyme in some F. oxysporum isolates produced four fragments (180bp, 160bp, 130 bp and 90 bp), so these isolates called as Fusarium redolens. MboI enzyme in others Fusarium oxysporum isolates produced two bands (320 bp and 190 bp). MboI enzyme had no restriction site in F. solani isolates. Conclusion: It seems that the use of rDNA-ITS RFLP marker is a suitable method to differentiate Fusarium species belonging to section Elegance from those of Martiella-Ventricosum sections. It is more efficient method for studying diversity within and between species in section Elegans.

Abstract Aim: Lizards are one of the most diverse and successful vertebrates in hot deserts of the world that biologists focus on their reproduction and gametogenesis.
Materials and Methods: This research is experimental study and the aim of that is to investigation of spermatogenesis steps and sperm production limit identification belongs spring three months. 10 male lizards (Laudakia Caucasia, Syn: Stellio caucasicus: Agamidae) were collected at the end of each month. After determination and morphological studies, their testes were dissected of animal body and prepared for histological studies using light microscopy. Quantitative data were analyzed using SPSS by ANOVA and Kruskal-Wallis tests (p < /em><0.05).
Results: Results showed that the spermatogenesis is started in the lizard from beginnings April and is gradually activated more with warming weather, As maximum spermatogenesis and mature spermatozoa production were observed in the end of June.
Conclusion: Based on histological evidences, spermatogenesis step is activated from early spring and is continued to the late of spring. Maximum sperm production is in June.
 

Abstract Aim: In this study, gene transfer efficiency of human lentiviral vector (HIV) into rat bone marrow stromal cells was compared with that of the feline lentiviral vector (FIV).
Material and Methods: To produce lentiviral vectors, a triple transfection with a shuttle vector, a backbone which carries either the GFP gene as a control or the NGF gene, and an envelope vector was performed using calcium-phosphate method in HEK-293T cell line. Supernatants of HEK-293T cell line containing the produced viruses were then exposed to bone marrow stromal cells (BMSCs) of the rat.
Results: While bone marrow stromal cells infected with HIV-NGF showed a relatively high immunoreactivity of NGF, those infected with FIV-NGF did not express NGF. Furthermore, BMSCs infected with FIV-NGF viruses containing antibiotic resistant gene could not survive in the culture media containing the antibiotic.
Conclusion: FIV based lentiviral vectors may not have a desirable efficacy for gene transfer into bone marrow stromal cells.
 

Abstract 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.
 

Abstract Aim:In the study, the effects of Bacillus thuringiensis parasporin were examined on mice cancer cell line (4T1).
Material and Methods: Purification of crystal toxins was done from the isolates of B. thuringiensis. Breast cancer cells line 4T1 were treated with activated toxins. Cytopathic effects were photographed. Crystal toxin with the most cytotoxicity was identified by using SDS-PAGE and also molecular method.
Results: From forty seven tested isolates of B. thuringiensis, E8 isolate shown the most cytotoxicity effect on 4T1 cell line. In addition, parasporin-4 was identified by using SDS-PAGE and PCR methods. Parasporin-4 disintegrated 4T1 cell line and it has cytolysin effects.
Conclusion: Our data suggest that, parasporin is cytolysin protein and has a specific site on the cell surface. This crystal protein can induce apoptosis in breast cancer cell line.

Abstract Aim: The present study aimed to investigate the effects of aqueous and alcoholic extracts of shallot on breast cancer cells compared to carboplatin and taxol treatments. Material and methods: Different concentrations of drugs taxol and carboplatin and aqueous and alcoholic extracts of shallots were prepared. Using Trypan blue and MTT, cytotoxicity on breast cancer cells, in different times and concentrations were evaluated. Utilized Hoechst and propidum iodide staining morphological changes and possible apoptosis of cells were determined. The data statistically analyzed using one-way analysis of variance and the mean level of P Results: The results indicate that shallot alcoholic extract has more inhibitory effects on cancer cells than aqueous extract at concentration range (0.01 to 0.05 g/L). Also, the simultaneous effect of shallot’s extract, accompaniment with carboplatin, caused enhancement in cytotoxicity of carboplatin. On the other hand, accompaniment of aqueous extract with taxol led to reduction of taxol cytotoxicity. Conclusions: This study showed aqueous and alcoholic extracts of shallots has cytotoxicity effects on breast cancer cells on rats affected by breast cancer. This plant is a herb which can be used against breast cancer  can be subjects for further research.

Abstract Aim: The effect of chitosan (low molecular weight) as elicitor was evaluated on silymarin production and biochemical factors in hairy root cultures of Silybum marianum. Material and methods: Different concentrations of chitosan (0, 5, 10, 20 and 30 mg/50 ml  of culture media) was added to hairy root cultures of S. marianum (30 days) and sampling  were carried out after 12, 24, 48, 72, 96 and  120 hours. The silymarin level, activity of guaiacol peroxidase and ascorbate peroxidase as well as concentration of H₂O₂ were determined in the samples. Results: The results indicated that the highest dry weight was in cultures treated with 10 mg /50 ml culture chitosan after 48 and 96 h. Silymarin production was significantly increased from 0.37 in control to 1.19 mg g^-1 dry weight in the treated samples, at which the increased level was 1.4-fold higher than the control. Guaiacol peroxidase was activated by chitosan and reaching a pick after 120 h (21.86 ∆OD g^-1FW min^-1) which was 2.2 fold higher than the control. The ascorbate peroxidase activity kept increasing until 96 h after elicitation (5.14 ∆ODg^-1FW). Conclusion: The results showed that the silymarin production has been promoted by chitosan (low molecular weight). Increased of enzymes activity occurs as a result of free radical elimination and represents a reaction to chitosan as a stress factor.  

Abstract Aim: The aim of this study was to evaluate the effects of rhizobial inoculation on the anatomical parameters of clover under SO₂ pollution.
Material and methods: 31 day-old plants (no-inoculated and inoculated with two strains of Rhizobium) exposed to the different concentrations of SO₂ (0 as a control, 0.5, 1, 1.5 and 2 ppm) for 5 consecutive days for 2 hours per day. The anatomical parameters of 40 day-old plants were Then investigated.
Results: Under high SO₂ concentrations (1, 1.5 and 2ppm), length of the palisade cells and diameter of the spongy cells decreased in both leaf surfaces. Stomatal density decreased in adaxial epidermis and increased in abaxial epidermis. Also, stomatal opening increased in adaxial epidermis and decreased in abaxial epidermis. Trichome density in both leaf surfaces significantly increased in 1.5 and 2ppm SO₂ compared with control plants. Under 2 ppm, trichome length increased significantly in both leaf surfaces. Inoculation of clover with two strains of Rhizobium significantly reduced the negative effects of high SO₂ concentration.
Conclusion: The results indicate the negative effects of high concentrations of SO₂ air pollution on the some anatomical parameters and the positive effects of bacterial inoculation on resistance to air SO₂ pollution stress.

Abstract Aim: The purpose of present research was extraction and cloning of tropinonereductase-II gene (tr-II) at antisense direction in pBI121 binary vector to provide transgenic plants with low rate of tropinonereductase-II enzyme and high production of scopolamine and hyoscyamine for future projects.
Material and methods:  Total RNA was extracted from Iranian native Hyoscyamusnigerroots, and the interest gene after cDNA synthesis and cloning at antisense direction in pBI121 binary vector, was transfered to Agrobacterium tumefacience. Accurate cloning was studied through 3 methods; enzymatic digestion, PCR and DNA sequencing. The bioinformatic characters of the gene were then surveyed.
Results: Three used methods confirmed true cloning in high efficiency. Nucleotide sequence of the gene revealed the 783 bp in length, encoding a polypeptide of 260 amino acid residues, with high similarity to that one registered in NCBI. The predicted molecular mass and isoelectric point of deduced polypeptide were 28437.3 Da and 5.46, respectively. Protein structures were not completely similar to those previously reported at PDB data base. Also, phylogenic study demonstrated that this gene belongs to the group I of TRs.
Conclusion: Due to successful cloning and high similarity of nucleotide and polypeptide sequences of gene with those recorded in world data bases; it is expected to get success in access to main purpose.
 
 

Abstract Aim: An endogenous repairman of the central nervous system is possible via neural stem cells. Some researchers have suggested the neural protective of some medicinal herbs such as; Urtica dioica. The purpose of this study is creating an oxidative condition in the neural stem cell cultures and then treated the cells with methanolic extract of nettle leaves in vitro.
Material and Methods: In this experimental study, the hippocampal neural stem cells extraction was performed by enzymatic digestion in newborn rats. The verification of these cells as nerve cells was carried out by the morphological and immunocytochemistry examinations. Before treatment, the cells were exposed for 24 hours to oxidative stress condition and then nettle leaf extract was added to the cell plates at of 2.5, 5, 10, 20 and 40 μg/ml and cells were maintained for 24 hours, separately. The final evaluation of the cell proliferation was performed by MTT viability test. The groups included; an experimental group that was treated with extract and the control group.
Results: Neural stem cells had neural morphology and expressed nestin marker. Proliferation percentage of the neural stem cells was higher in the treatment groups than the control group. In addition, MTT assay results showed that the percentage of live cells in the treated groups increased, as the proliferation of the neural stem cells significantly increased (p < 0/05) at 20μg/ml.
Conclusion: Methanolic extract of nettle leaf have neuroprotective effects and could adjust oxidative stress condition in vitro. It was able to improve the dysfunction of the central nervous system, following the production of free radicals.
 

Abstract Aim: Effects of two nanoparticles were investigated on the expression of STR, DAT and D4H genes in periwinkle.
Material and Methods: For this purpose, concentrations of 0.5, 0.75 and 1 mg per liter of cobalt and zinc oxide nanoparticles were used. Sampling was carried out at 0, 8, 24 and 48 hours after treatment.
Results: Gene expression analysis was performed by the SQ-RT-PCR method. In general, both elicitors influenced gene expression. But, the effects of zinc oxide nanoparticles on the gene expression were more pronounced than cobalt oxide nanoparticles. The highest expression of STR and D4H genes were occurred in 0.5 mg per liter of zinc oxide nanoparticles after 8 hours treatment, while in the case of DAT gene, it occurred in concentration of 1 mg per liter of this nanoparticle. Moreover, Cobalt oxide nanoparticles in most of the studied concentrations and time intervals caused decreases in gene expression.
Conclusion: Both nanoparticles had significant effects on gene expression of vinblastine and vincristine pathways.

Abstract Aim: In this study, the extract of earthworm (Eisenia foetida) (in solvent: olive oil and vaselline) was applied as a novel healing stimulator on suture wound in goldfish (Carassius auratus).  
Material and Methods: Groups were: negative control (Intact skin), positive control (suture wound with phenytoin), pseudo-control (sham: suture wound with solvent), and experimental (suture wound with 10 mg earthworm extract). A 2 cm longitudinal cut was made from head to tail, and then suture was applied followed by daily treatments. After 14 days, blood samples were collected. Macroscopic and microscopic observations were also done. Meanwhile, the content of lipid peroxidation (LPO/MDA) was measured in fish blood sera.
Results: surface and microscopic observations of wounds revealed a better result for earthworm extract treatment in suture wound healing when compared with phenytoin. MDA level in goldfish blood sera was lesser in earthworm extract-treated groups compared to other groups.
Conclusion: Earthworm extract is a new candidate in aquatic wound healing. Some factors of the extract may enter the blood and ultimately activate the antioxidant systems leading to decrease the content of LPO in fish blood sera.