Cell and Tissue Journal

Abstract Aim: Cadmium is one of the air pollutants and a part of diesel exhaust particles. The aim of this study was determination of the effects of cadmium on developmental process and protein pattern of anther and pollen grains in Petunia hybridia that is a common decorative plant in landscapes and parks.
Material and methods: Petunia plants were grown in experimental pots and treated with the different concentrations of CdCl₂ as spray. Flowers and young buds were collected, fixed in FAA₇₀ (formalin: acetic acid: ethanol with a ratio of 2: 1: 17) and subjected to developmental studies. Pollen grains of different groups were also collected and their proteins were studied using electrophoresis method. Statistical analysis was done using statistical software SAS (version 9.1) and one-way ANOVA analysis.
Results: The developmental process of anther and pollen grains in control plants were followed from the pattern of dicotyledonous plants, but some changes and abnormalities were observed in Cd-treated plants. Irregularity in anther wall, formation of wrinkled and vacuolated pollen grains, decreasing of pollen viability, damaged exine and formation of new protein bands are the effect of Cd treatment.  
Conclusion:Cadmium, as an atmospheric pollutant, influences developmental stages of anther and pollen grains. It causes to form different abnormalities and reduction of pollen viability. Formation of new proteins, detoxifying proteins, is a response to pollution stress. 

Abstract Aim: As there was no studies available regarding the morphological structure of skin of (Barbus grypus) therefore this research work was performed to help the other studies such as histopathology of the fish.
Material and Methods: Skin of the different part of the head of 20 Barbus grypus with 5mm thickness were removed and fixed in bouin solution. The routine procedures for preparation of tissues were followed. For transmission electron microscopic study, the samples after primary and post-fixation were dehydrated and embedded in resin. Then, thin sections 50 µm were prepared and stained with uranyl acetate.
Results: according to histomorphological examination, skin of head, lip and barbel was made of epidermis and dermis which were located on a loose connective tissue called hypoderm. In histometric study, skin of studied area, was divided into different regions in (Barbus grypus)based on cells distribution. In the micrograph of TEM, goblet cells containing many mucus granule, spinous and club cells with hyper chromatin nucleus were observed, which presence of hyper chromatin nucleus was more obvious in the club cells. 
Conclusion: The light and TEM studies on the skin in studied region of (Barbus grypus), showed there was differences and some similarity with the other reported species. 
 

Abstract -
Aim: This study was carried out to investigate the application of different magnetic intensities on the sperm freezing medium.
Material and methods: In this experiment, the magnetic field induction of 0, 2000, 4000 and 6000 Gauss was used to determine the optimum intensity during the freeze-thawing process. Semen samples were collected from 10 pieces of 24-week-old Ross 308 broiler chickens using abdominal massage technique. In this study, the experimental treatments were: 1) ordinary diluent, 2) G2000-intensive magnetic diluent, 3) G4000-intensive magnetic diluent, and G6000-intensive magnetic diluent. The used diluent in this study was the Lake diluent. This diluent was mixed with 1% soy lecithin and 5% glycerol, and then on the basis of the experimental treatments, they encountered different magnetic intensities. Diluted sperm was frozen based on each treatment, after cooling in a 0.25 ml straws. After freezing some sperm parameters such as total and progressive motilities, morphology, membrane integrity, MDA production, apoptosis status and mitochondrial activity were evaluated.
Results: Results showed that different magnetic fields had no significant effect on total and progressive mobilities, membrane integrity, morphology, lipid peroxidation, acrosome integrity and apoptosis mechanisms of rooster sperm. Just mitochondrial membrane potential in the control group was higher than other groups (60.5%, p < 0.05).
Conclusion: It seems that exposure of rooster semen cryopreservation media with magnetic field cannot be efficient for quality of sperm after thawing.

Abstract Aim: The aim of this study was to investigate the preventive role of vitaminE (Vit.E) on spermatogenesis indexes in rats following exposure to Bisphenol A. Materialand and Methods: Adult male wistar rats with the mean body weight of 231±10g were randomly divided into 4 groups (n=6): control, BPA (250mg/kg/day), Vit.E (150mg/kg/day) and BPA + Vit.E.  Oral treatment was performed three times a week till 56 days. At the end of the treatment, the rats were killed and their left testis were weighed, fixed and stained using Heidenhain's Azan  methed. Histological and morphometrical analysis of spermatogenesis was carried out. Data were analyzed using One Way ANOVA and  the means were considered significantly different  at p < strong>Results: Testis weight (p < 0.01) significantly decreased in the BPA group compared to the control. A significant decrease (p < 0.001) in the mean diameter of seminiferous tubules and the germinal epithelium thickness and the indexes of tubular differentiation, spermiogenesis and meiosis were also observed in the testicular tissue of rats treated with BPA compared to the control ones. In the BPA +Vit.E group, the mentioned parameters increased significantly to the control level. Conclusion: Vitamin E can compensate for the undesired effects of BPA on spermatogenesis and therefore could be considered as a therapeutic supplement in the case of BPA toxicity.  

Abstract Aim: In this study the toxic effect of the three anti hypertensive drugs (atenolol, propranolol and diltiazem) on hepatocytes were investigated.
Materials and Methods: The 8 weeks old adult male NMRI mice were fed with or without the drugs for 1 month. Mice were divided into 4 groups that received atenolol (50 mg/kg/daily), propranolol (80 mg/kg/daily), and diltiazem (180 mg/kg/daily). Food access was stoped 12 to 15 h before the mice were sacrificed and processed for light and transmission electron microscopic evaluation. In addition modified Hepatitis Activity Index was computed too.
Results: In the control group no pathological sign was observed. Main histological changes such as portal infiltration of lymphocytes and piecemeal necrosis in all three experimental groups as well as expanded necrosis in proparanolol group were seen. In addition electron microscopic imaging indicated an increase in the level of free ribosomes and glycogen droplets in the experimental groups. Also in the mice treated with propranolol group mitochondria membrane disruption were seen, whereas in the group treated with diltiazem, the mitochondrial cristae and endoplasmic reticulum Disintegration could be observed.
Conclusion: Chronic oral administration of atenolol, propranolol and diltiazem in male mice caused hepatotoxicity. The most pathological effects were observed in the group treated with diltiazem, though electrone microscopic imaging indicated the propranolol and dilitiazem were more toxic than the other one.  
 

Abstract Aim: Varicocele is the most common causes of male sterility that is caused progressive testicular damage over time. In this study sperm parameters and their DNA integrity was compared in varicocele and fertile people.
Material and Methods: The study performed on 55 grades II and III Varicocele people and 25 fertiles. Semen parameters (sperm count, motility and morphology) and sperm DNA health were assessed using TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling). Data were analyzed by SPSS software.
Result: Varicoceles semen parameters consist of sperm count, motility and morphology had significantly low quality in comparison with fertile people. Also damaged DNA sperms percentage of Varicocele people (14.34%±1.43) was significantly increased compared to fertile people (9.2%±1.36). Sperm parameters and DNA damage percentage correlation showed that there is a reversed correlation between DNA damage percentage and sperm density and significant and positive correlations with sperm abnormal morphology.
Conclusion: Testicular temperature increasing in Varicocele people, caused to low quality of sperm parameters and also high sperm DNA damages. Therefore, Varicocele people are confronted with spermatogenesis process deficiency.
 
 

Abstract Aim: This research was conducted to investigate changes in single starnded DNA preferring nuclease (SSPN) activity, protein and chlorophyll contents in barley, under salt stress.

Material and Methods: Tolerant (Sahra) and sensitive (Rayhan) barley cultivars were used in a complete random plot with a factorial design, using 3 replicates at 0 and 120 mM NaCl concentrations. Nuclease activity on ssDNA was measured at pH 5.6 and 7.0 in the presence of 1 mM of one of the following divalent cations: Cu²⁺, Zn²⁺, Mg²⁺, Ca²⁺ and EDTA, by spectrophotometry and native-PAGE assays.

Results: The highest enzyme activity was observed in the presence of Ca²⁺ at 120 mM NaCl and pH 7.0 and the lowest enzyme activity was observed in the presence of Cu²⁺ at 0 mM NaCl and pH 5.6. EDTA inhibited SSPN activity. In the gel assay, 5 enzymes were revealed, amongst which 59, 47 and 43 kD enzyme at both pHs and 34 and 41 kD enzymes were revealed only at pH 7.0. Na+ content, K+/Na+ ratio and chlorophyll and protein contents decreased significantly upon salt stress in both cultivars.

Conclusion: Salt stress caused an increase in the nuclease activity and a decrease in protein and chlorophyll contents in both cultivars. However, these changes were significantly lower in the tolerant cultivar, compared to the sensitive one

Abstract Aim: The purpose of this study was to investigate the effects of benfotiamine treatment of hyperglycemic rats on viability of adipose tissue- derived mesenchymal stem cells on sciatic nerve acellular scaffolds.
Material and method: After induction of hyperglycemia (STZ) and treatment with benfotiamine segments for 4 and 8 weeks from the middle part of the sciatic nerves were  decellularized using Sandell method and seeded with  mesenchymal stem cells derived from adipose tissue. after 8 days of culture cells viability in the culture medium was evaluated by MTT assay and  adhesion of the cells to the scaffold was examined by scanning electron microscopy (SEM).
Results: In comparison with intact group, cell viability of untreated hyperglycemic rats was significantly decreased while, there was no significant difference between benfotiamine  treated group and intact. Results of electron microscopy confirmed adhesion of  the cells on the scaffolds.
Conclusion: In hyperglycemic condition it is likely that glycosylation of ECM constituents and the production of AGEs decrease the inducible effects of ECM on the level of cell viability and probably cell adhesion to the scaffolds. It seems that benfotiamine treatment of hyperglycemic rats by reduction of glycation and AGEs production may prevent the structural changes of the ECM.
 

Abstract Aim: Sulfur mustard (SM) is an alkylating agent which targets skin, eyes and respiratory systems. Also, an insertion/deletion polymorphism in angiotensin converting enzyme (ACE) gene has been studied in relation to cardiovascular diseases and lung fibrosis. In pulmonary fibrosis, D-allele frequency is higher than in comparison with control. So, the aim of this research was studying relashionship between ACE genotype and late respiratory complications of mustard gas in Kermanshah Zardeh Village chemical exposed people.
Material and Methods: Blood samples of thirty-four people of Kermanshah Zardeh Village, whose were exposed to mustard gas as a case study and thirty people of Eslam Abad Gharb in Kermanshah Province as control, were taken. Further information about the existence of respiratory, skin and ocular delayed effect of exposure to the mustard gas, were collected by questionnaires. ACE genotype was determined using polymerase chain reaction amplification (PCR) and subsequent agarose gel electrophoresis.
Results: Incidence of respiratory, skin and eye complications were 52.9%, 50% and 44.1% respectively. Genotype frequencies for the three genotypes DD, ID and II in case group of people with pulmonary complications were 0.5, 0.44 and 0.06 respectively, in people without this complications 0.12, 0.69 and 0.19 and in the control group were 0.3, 0.53 and 0.17 respectively. It was found that the DD genotype in patients with respiratory symptoms were higher than those without complications ( =6.22, p=0.045, df=2).
Conclusion: The results showed that DD genotype of ACE gene increases the risk of respiratory complications of mustard gas.

Abstract Aim: In this study, sodium nitroprusside effect as a producer of NO on growth, antioxidant enzyme activities and some physiological parameters in in vitro cultured Melissa officinalis seedlings were studied.
Material and methods: After establishment of Melissa officinalis seedling culture, seedlings were treated with different concentrations of sodium nitroprusside (0, 25, 50 and 100 µM). Then, they were harvested for growth, protein content, secondary metabolites, lipid peroxidation level and antioxidant enzymes activities analysis after 10 days.
Results: Results showed that growth and protein content decreased with increasing concentration of sodium nitroprusside. Phenolic compounds amount, total flavonoids and anthocyanins increased by sodium nitroprusside. Also Malondialdehyde content as lipid peroxidation product was significantly increased. In response to oxidative stress, chatalase and peroxidase enzymes activities were respectively increased at 50 and 25 µM sodium nitroprusside while enzymes activities showed decreasing at 100 µM.
 Conclusion: Overall, the results indicate that sodium nitroprusside as NO producer damage the cell membrane. Therefore reduced Melissa officinalis seedling growth and caused oxidative stress in them. To overcome produced free radical, secondary metabolites amounts and antioxidant enzymes activities were increased.

Abstract Aim: This study was conducted to determine the neuroprotective effects of mint extract on alpha motor neuron degeneration at the anterior horn of the spinal cord after sciatic nerve compression in rats.
Material and Methods: In this study, 30 male Wistar rats weighing 200-250 g were randomly divided into 5 groups including control, compression and treatment groups of 50,75 and 100 mg.  In order to induce the compression, sciatic nerve was undergone to compress by using locking- scissors for 60 seconds. Hydroalcoholic extract of mint was injected intraperitoneally during the first and second weeks after the compression. After 28 days, rats were undergone by the perfusion method and after sampling the lumbar spinal cord, neuronal density was calculated by using dissector and stereological methods and the findings were compared together.
Results: a significant decrease was observed in compression group compared to the control for the neuronal density and also a significant increase was seen in treatment groups rather than the compression group (p

Abstract Aim: Regarding economic, medicinal, food and decorative importance of Artemisia genus,this study was carried out to determination of intraspecific diversity in the Artemisia incana populations collected from East Azerbaijan. 
Materials and methods: To study of intraspecific diversity in A. incana in East-Azerbaijan D.S.S. method was used. Six special habitats of this population were recognized. Seeds of populations as well asand floristic-ecologic data were collected from each special habitat. Seed storage proteins were subjected for electrophoresis studies.
Results:  In survey of all special habitats, 54 species were distinguished as associated species. Analysis of floristic data (Floristic compositions of each special habitat as floristic marker) led to identification of 4 separate groups that was indicated the existence of intraspecific diversity in this species. Analysis of ecologic data was also confirmed the groupment. The analysis of seed storage protein electrophoresis banding pattern showed three quite obvious groups.
Conclusion: In the survey of electrophoresis pattern, existence of differences regarding number and density of the protein bands indicating existence of intraspecific diversity in the populations of           A. incana. Therefore in this species, the grouping that introduced with floristic marker, confirmed also with ecological and electrophoresis markers. This means that floristic grouping can only be used as a cost-effective and efficient method for studying of intraspecific diversity.

Abstract Aim: In this study, effects of Rhizobium on alfalfa plant growth under the SO₂ pollution were evaluated
Material and methods: 35 days plants (non-inoculated and inoculated with native or standard Rhizobium) exposed to the different concentration of SO₂ (0, 0.5, 1, 1.5 and 2ppm) for 6 consecutive days for 2 hours per day.
Results: Results indicated significant decrease of Leaf area, stem length, root depth and fresh and dry weight of leaf, stem and root under high concentration of SO₂. Number of leaves at all concentrations showed no significant change. Inoculation of alfalfa with Rhizobium significantly reduced the negative effects of high concentration of SO₂. In this study, positive effects of native bacteria were impressive.
Conclusion: The results indicate the harmful effects of high concentrations of SO₂ pollution on growth of plants and the positive effects of bacterial inoculation on resistance to air SO₂ pollution stress.
 

Abstract Aim: In the current study, the protective effect of quercetin (as an antioxidant) was investigated on the ovarian tissue and fertility of female rats exposed to cyclophosphamide.
Material and Methods: 24 Wistar rats were divided into four six-membered groups. The first group was treated with cyclophosphamide (30 mg/kg) and the second group was treated with tween solution. The third and fourth groups administered quercetin (50 and 100 mg/kg) with cyclophosphamide (30mg/kg) intraperitonealy for 30 days. The left ovaries of the rats were removed and after sectioning and staining using Hematoxylin-Eosin were examined using light microscopy. In addition, 16 rat were treated in the same way in 4 groups (one male and three female rat in each group), then the mice mating. After birth, growth indexes were studied. All data were analyzed using one-way ANOVA.
Results: Both doses of quercetin significantly increased the number of primordial follicles and diameter of the primary follicles, the number of blood vessels, the number of offspring and their growth indexes, while decreased the number of atretic graffian follicles,. In addition, a significant increase in the diameter of primordial follicles was observed in rats received 100 mg/Kg quercetin compared to rats received only cyclophosphamide.
Conclusion: Quercetin can reduce the side effects of cyclophosphamide on ovarian tissue and improve fertility indexes.
 

Abstract Aim: 7-bromoindirubin-3'-oxime (7-BIO) is a well-known glycogen synthase kinase-3β (GSK3β) inhibitor with anticancer effects on different cancer cell lines. However, due to its low water solubility, absorption and high gastrointestinal toxicity, it has been inapplicable in clinics so far. In the present study, bovine serum albumin nanoparticles (BSA NPs) were prepared as drug carriers.
 Material and methods: to be conjugated with 7-BIO to increase the effectiveness and reduce the toxicity. The nanoparticles were synthesized and conjugated with 7-BIO and their sizes were analysed by using scanning electron microscopy. Also, the viability and apoptosis in the cells treated with 7-BIO conjugated nanoparticles were examined by MTT assay and acridine orange/ethidium bromide staining (AO/EB), respectively. The significance of the data variation was evaluated by using ANOVA test.
Results: 10 mg/ml albumin led to spherical nanoparticles (NPs) of 89.42±7 nm in diameter with mono dispersity. MTT assay showed that 7Bio-loaded BSA NPs had a higher toxic effect on MCF-7 cell line compared to the free 7-BIO. Also, AO/EB staining showed that the apoptosis was induced by 7-BIO treatments. 7-BIO is known to specifically inhibit GSK3β phosphorylate GSK3β, which was also evident in our treated breast cancer cell line MCF-7 both with free and loaded BSA NPs 7Bio.
 Conclusion: The albumin nanoparticles conjugated with the GSK3 inhibitor effectively decreased the viability of the proliferating breast cancer cell line MCF-7, pointing at its possible clinical impact in future.
 
 

Abstract Aim: In the present study the effects of central injection of Orexin A on the expression of TSPO gene in the ovaries of pubertal androgenized female rats were investigated. Material and Methods: 24 neonatal female rats were androgenized on the third day after birth by subcutaneous injection of 50µg TP. Six neonatal female rats in one group were considered as controls. After puberty, the animals in 5 groups (n=6 in each group) received central injections of saline, different doses of Orexin A (2, 4 or 8µg). The ovaries were removed bilaterally and frozen. TSPO gene expression levels was determined by semi quantitative RT-PCR. Results: The mRNA levels of TSPO increased significantly in the ovaries of the androgenized rats compared to controls. Orexin A injections decreased significantly TSPO gene expression compared to the androgenized rats (P <0.05). Conclusion: Androgens may stimulate TSPO gene expression in the ovaries. Orexin A may exert inhibitory effects on reproductive axis partly via reducing the expression of genes involved in the steroidogenesis.

Abstract Aim: The aim of this study was to use carbon nanotube incorporated electrospun chitosan/polyvinyl alcohol (CS/PVA/CNT) to induce neural differentiation of the stem cells for potential neural tissue engineering applications.
Material and Methods: P19 embryonal carcinoma (EC) stem cells were cultured on the CS/PVA/CNT scaffold to induce neuronal differentiation. Cresyl violet staining was used to investigate neuronal phenotype and immunoflourescence technique was carried out to evaluate expression of β-tubulin, a neural specific marker.
Results: Cresyl violet staining confirmed the neuronal morphology of the differentiated cells. These cells were immunoreactive to neuron specific marker, β-tubulin.
Conclusion: the findings suggested the potential use of combined tissue engineering and stem cell therapy to improve deficits in neurodegenerative diseases.
 

Abstract Histological and Electron Microscopy Study of the Intestinal Tissue of Rainbow Trout (Oncorhynchusmykiss): an In Vitro Model

Abstract Aim: Evaluation ofprotective effectsof Ephedra pachyclada on carbon tetrachloride- induced acuteliverinjury.
Material and method: In this study,initially wedesignedan experiment to achievea standardanimalmodel, in which 30mice were divided into three control, sham andmodelgroups. In themodel group the animal received2ml/kgcarbon tetrachloride50 %( 1:1 in sterile olive oil)intraperitoneally. 72 hourslater,bloodand liversamples were collected and analyzed using biochemicalas well ashistologicalstudies. Afterensuring theeffectiveness of the modelinduction, in the secondtrial, 40 micewere divided into4 groups including controlandthreeexperimentalgroups. Induction ofacuteliverinjury was performedsame asprevious on experimentalgroups, whichtwoofthemweretreatedwith Ephedra pachycladaextract(140, 1400 mg / kg) daily. Bloodand liversampleswere collected96 hafter carbon tetrachloride injection then serumactivity of Alanine amino transferase (ALT), Aspartateamino transferase (AST) andnecrosisas well asinflammationin the liverwere studied.
Result: Treatmentwith Ephedra pachycladaextract significantly decreased the activity of ALTandAST in the serum as well as thenecrosisandinflammation in liver tissue. In addition the survival rate was increased in the animal receivingcarbon tetrachloride.
Conclusion: Results of this researchsuggestthat Ephedra pachycladaprotects the liver against acute liver injuryinduced by carbon tetrachloridethroughinhibition ofoxidative stressandsuppressing theinflammation.
 
 

Abstract Aim:This study was performed to evaluate the cytotoxic effect of Ocimum basilicum and Impatiens walleriana aqueous extract on the gastric cancer of AGS cell line.
Material and methods: FRAP method was used to measure the antioxidant activity of Ocimum basilicum and Ipatiems walleriana extracts. After preparation of O. basilicum and I. walleriana aqueous extract, we measured the anticancer effect on growth of AGS cancer cells at 24, 48 and 72 hours by MTT.
Results: The highest antioxidant activity of O. basilicum and I. walleriana  was measured by FRAP method for water solvent and water bath extraction method, which in O. basilicum was

0.778 ± 0.11 mM and in I.walleriana  1.30 ± 0.03 mM. Based on the obtained data of the MTT test, the highest toxicity of the extract occurs 72 hours after adding the extract to the cells. The lowest amount of IC50 of the aqueous extract of O. basilicum was measured at 0.012 ± 0.002 mg / ml and in I. walleriana extract was 2.87 ± 0.001 mg / ml., which had a significant difference ( p <0.05).
Conclusion: The results of the anticancer study of the aqueous extract of O. basilicum and I. walleriana indicate that these plants have a cytotoxicity effect in the higher concentrations and need a long duration for their effects.
 

Abstract Aim: One of the most important infraspecific variations is differences of tissues and cells in plant that occur between various populations of the same species for adaption with environmental factors. These variations can be consider as a starter for speciation.
Material and methods: In the present study, leaf anatomical traits of four Nepeta heliotropifolia populationswere examine. Three flowering stems were collected from each population, and from each stem a mature intact leaf was elected. Leaves were fixed in fixative and then transferred to ethanol. Hand sections were decolorized, double stained and their anatomical structure were studied using light microscopy. The MVSP and SPSS soft wares were used for statistical analyses. 
Results: In total, twenty two qualitative and quantitative anatomical characteristics were studied. Qualitative features were stable between populations, while ANOVA test showed significant differences for most of the studied variables. In addition, significant correlations were found between morphological characters with each other and ecological factors of habitats. The studied populations clustered separately in UPGMA tree, PCA and PCO plots.
Conclusion: Environmental factors have effects on tissues and cells of plant and create infraspecific variations. They have more effects on quantitative variables compared with qualitative ones. Leaf anatomical variations of populations have no relationship with distances between the populations, but similarity or difference of ecological factors is a very important factor in creating similarity or difference between populations. The pattern of population’s arrangement in the created tree and plots approved it. 
 

Abstract Aim: The purpose of this study was to consider the effect of mild oxidative stress period on the frozen-thawed bull semen performance.
Material and Methods: In this study, 120 minutes was considered for equilibration of sperm before freezing. One µm of Nitric Oxide (NO) at the 0, 45, 90 and 120 cooling period was added to the diluant before cryopreservation. Sperm motion parameters, plasma membrane integrity, acrosome integrity, viability, apoptosis and mitochondria activity were assessed.
Results: Induction of oxidative stress with nitric oxide in the T0 and T45 groups led to significant improvement of total (88.4 ± 2.8, 84.7 ± 2.7) and progressive motility (50.4 ± 2.5, 49.5 ± 2.5) when compared to other groups. Percentage of plasma membrane integrity and viability were not different in T0 (74.4 ± 2.9 and 85.6 ± 2.9) and T45 (75.6 ± 2.9 and 84.6 ± 2.3) groups, where as it was significantly higher when compared with T90 (63.6 ± 2.9 and 69.6 ± 2.3) and T120 (61.5 ± 2.9 and 54 ± 2.3). Moreover, the highest significant percentage of spermatozoa with active mitochondria was observed in the T0 (82.5 ± 3.1) when compared with T45 (65.7 ± 3.1), T90 (42 ± 3.1) and T120 (43 ± 3.1). Acrosome integrity and linearity of sperm were not affected by the oxidative stress treatment time.
Conclusion: It seems that applying oxidative stress using 1 µM NO would improve post-thawed bull sperm quality at the T0 and T45 time of cooling.

Abstract Aim: Papaver somniferum remains the sole commercial source for several pharmaceutical benzophenanthridine alkaloids from benzylisoquinoline branch alkaloids, which includes the narcotic analgesics codeine and the semi-synthetic drugs such as; oxycodone, buprenorphine and naltrexone. Although, most of the biosynthetic pathways genes of this alkaloid have been identified, the post-transcriptional regulation of these alkaloids pathway has not been completely determined. Virus induced gene silencing (VIGS) is a method for fast functional genomics, and in this study, this technique was used to investigate the silencing one of the most important genes in this alkaloid pathway.
Materials and Methods: In the current research VIGS technique was used for systematic reduction in the level of genes expression involves in benzylisoquinoline alkaloids pathway. For silencing of codeinone reductase gene, pTRV vector was used for cloning. The specific silencing of COR gene was evaluated in 2-3 weeks old leaves of Papaver somniferum using agro-infiltration method.
Results: Result of cloning was confirmed by using of different molecular methods such as enzyme digestion and also PCR. PCR technique was used for confirmation of transgenic plants in some transformed plants. Using of semi-quantitative PCR and real-time PCR showed that the level of reduction in transcription of COR gene was about 89 percent.
Conclusion: The obtained results confirmed that by application of RNA interference method, the level of COR gene expression was significantly reduced compared with control.

Abstract Aim: In this research morphological, cell culture, immunochemical and inductive properties of rat’s claw dermal cells were investigated. Material and Methods: First, the toes and fingers of rat were amputated at the point of the last joint and the amputated parts were processed for histological studies. Moreover, the dermal tissue located beneath the claw’s plate was dissected and cultured in vitro. Cultured cells at passage two or three were transferred on the slide and treated with α-smooth muscle actin (ASMA) antibody. Finally the cultured cells were stained with DiI and implanted into the vibrissae semi-follicles and ears. Results: The rat’s claw dermal cells in terms of morphological, cell culture and expression of ASMA demonstrated similarity with dermal papilla cells of hair follicles. These cells showed a bipolar spindle-shaped morphology having an extracellular matrix rich in glycosaminoglycans. Majority of the cells expressed ASMA in cytoplasm with a higher intensity in lamelopodia.  However, with respect to hair follicular dermal papilla cells, the claw dermal cells were not able to induce an ectopic epidermis to form the skin appendage.  Conclusion: The inductive capability which has been seen in hair follicle’s dermal papilla cells can not be generalized to dermal cells present in other skin appendages, including claw. Based on the results presented here, the inductive factors exist in hair follicle dermal papilla cells is not expressed in the claw dermal cells.

Abstract Aim: The aim of this project is finding a relationship between body characters of Hirudo orientalis with testis sac and spermatic follicles structure in order to find a relationship between these characters and sexual maturity, regarding farming goals.
Material and methods: 30 individuals of H. orientalis were collected From 3 ponds located in Gonbad-é-Kavous city in May, July and November. Body length, width and weight, Testis sac diameter, testis sac diameter to body diameter ratio and spermatic follicle numbers obtained for each sample. For understanding starvation effect on testis sac, 20 individuals of collected leeches were starved for 120 days. Data were analyzed using ANOVA test and SPSS 16 software.
Results: Body length, width and weight, testis sac diameter, testis sac diameter to body diameter ratio and spermatic follicle numbers showed significantly reduction in May comparing with July and November (p < /em><0.05). Mature spermatic follicles were observed in samples with more than 3 gr weight, >10.3 cm length and >0.79 cm diameter in July and November. All of studied characters were reduced with the exception of body length and testis sac diameter to body diameter ratio in starved samples (p < /em><0.05).
Conclusion: This study showed that testis sac structure and spermatic follicle numbers can directly related to body weight and diameter in H. orientalis but not to body length.