Cell and Tissue Journal

Abstract Aim: During recent years many studies have suggested some developmental roles for embryonic cerebrospinal fluid. Here we examined the effect of embryonic cerebrospinal fluid on proliferation and self-renewal of embryonic ventricular zone derived neurosphere.
Material and Methods: Cortex from 15.5 day old embryonic Wistar rat dissected out and enzymaticaly dissociated to form single cell suspension. Cell suspension seeded in DMEM/F12 medium supplemented with N2 and mitogenes (10ng.ml^-1 EGF and 20ng.ml^-1 bFGF). The CSF-treated culture from different embryonic ages (E16, E18, and E20) in 10/100 ratio (v/v) was considered as experimental groups. Neurospheres number was counted for proliferation assay and cell viability evaluated with MTT assay. The data were analyzed with One-way ANOVA with turkey’s post hoc test.
Results: Embryonic cerebrospinal fluid (e-CSF) enhanced neurosphere number. Also, embryonic cerebrospinal fluid (e-CSF) enhanced cell viability and growth.
Conclusion: Embryonic cerebrospinal fluid (e-CSF) enhanced proliferation and viability of neural progenitor cells in age dependent manner. Moreover, this fluid play important role in establishment of neuroprogenitor cells self-renewal.

Abstract Aim: In this article, the ability of p < sup>32 radioactive drugs for treatment and control of breast cancer in rat model and also verifying of distribution of drug in rat organs are investigated.
Material and Methods: 10 to 20 mg per kg of DMBA gavaged to SD strain rats. Tumors 15-10 mm in diameter were selected for testing. P32 radioactive drugs injected to developed tumors with interval 3, 5 and 7 days. The distribution of the P32 radioactive drugs in the body of rat were analyzed by NI detector. After the treatment period, animals were anesthetized and the tumor is completely removed under sterile conditions and placed in 10% formalin. Sections of the tumors were obtained and processed using Tissue Processor. After tissue processing and sectioning, the samples were stained with hematoxylin - eosin were examined.
Results: The results obtained from the examining histological sections prepared from tumor tissue confirmed that, the pathology of tumors induced by DMBA in rats used in this study is very similar to human mammary tumors. Five days after injection, we could find tumor's cells and lymph ducts are covered by residual epithelial cells. Many of the cells are separated from the basement membrane and collapsed into the tubular cavity. Rest of the cells has small nuclei, with chromatin condensation mode and cytoplasm was compact and dense. In radius of about 7-8 mm around the site of injection the tumor's cells were necrosis and formed a cavity, inside of tumor.
Conclusions: These observations showed further penetration and influences of radiation emitted from the phosphor 32 in the tumor tissue based on our injection pattern.
Therefore if given the proper distance of injection within the tumor, destroying the tumor's ability to be on time will be shorter.
 

Abstract Aim: The aim of this research was to investigate the effects of different concentrations of Azadirachta indica )neem( aqueous extract on the expression of the encoding genes of the two phenylalanine ammonialysis and peroxidase enzymes in tomato (Matin and Early Urbana cultivars) in the interaction with root knot nematode caused by Meloidogyne javanica.
Material and methods: The tomato seedlings were inoculated in the 4-6 leafed stages with different concentrations of the neem extract and then incubated with the pathogen. The plants sampling were at six time points (0, 12, 24, 96, 192 and 288 hours) after pathogen inoculation. The gene expression of the defense enzymes was investigated using the Real-time PCR techniques.
Results: In both cultivars, the gene expression of POX and PAL enzymes in the combined treatment of the pathogen nematode and the aqueous neem extract had increasing trend with significant difference with the control. In Matin cultivar, the increase of the gene expression was higher and faster than of Early Urbana cultivar, so that the expression of PAL and POX genes in this cultivar was 18.2 and 14.5 times more than the control. In the early Urbana cultivar, the highest rate of PAL gene expression was observed at 19.2 hours and was 7.5 times, and in the case of POX gene at 96 hours and 3.9 times when it was compared with control. In greenhouse experiments, the nematode pathogenicity indexes showed a significant decrease compared with the Early Urbana cultivar.
Conclusion: According to the results, in order to protect the plant against the invasions of the patients, the expression of the defensive genes should be increased at the earliest stages after the inoculation of the patient. Early increase in expression of genes in the seedlings treated with neem extract seems to be a justifiable phenomenon in the plant resistance to the pathogenic agent.
 

Abstract Aim: Due to the role of silver nanoparticles on elevation of reactive oxygen species, and high sensitivity of spermatozoa to these agents, it is possible that silver nanoparticles reduce the fertility potential of spermatozoa. The aim of present study was to evaluate the effects of silver nanoparticles on sperm parameters and the concentration of reactive oxygen species in mouse serum and seminal fluid.
Material and Methods: In this experimental study, 24 adult male Syrian mice were divided into a control and three experimental groups. The experimental groups received silver nanoparticles orally at different doses (0.05, 0.1 and 0.2 ml orally) for 5 weeks. Then blood samples were taken from hearts to examine the Malondialdehyde (MDA) concentration by spectrophotometry. In addition, to analyze the sperm parameter cauda epididymis was dissected and swim-out spermatozoa were analyzed.
Results: The 3rd experimental group (highest dose) significantly showed the lowest mean of sperm count (16±2.6), the lowest percentage of rapid progressive motile spermatozoa (18.16±5.45), the highest rates of abnormal spermatozoa (35.83±4.4) and the highest concentrations of MDA in serum and seminal plasma compared to the control and other experimental groups.
Conclusion: The silver nanoparticles as a factor which increase the effects of free radicals and reactive oxygen species in serum and seminal fluid caused reduction of sperm parameters.

Abstract Aim: The purpose of this study was to investigate the effect of ultrasound on growth and some physiological parameters in cultures of unicellular alga Dunaliella salina. Material and Methods: Ultrasound waves were applied with a frequency of 40 kHz, and 5W/Cm3 power, at the fourteenth day of subculture in a completely randomized design with 3 replications. Ultrasound exposure times to cells were 0, 2.5, 5 and 10 minutes. The parameters measured were: cell growth, total protein content, photosynthetic pigments, antioxidant potential, membrane lipid peroxidation, amount of phenolic compounds, flavonoids, anthocyanins, soluble sugars, beta-carotene and glycerol. Results: The results showed, due to increase of ultrasound irradiation time, cell growth and photosynthetic pigments were decreased. In contrast, total protein content, antioxidant potential, membrane lipid peroxidation, phenolic compounds, flavonoids, anthocyanins, beta-carotene and glycerol were increased. Maximum amount of beta-carotene (12.3 mg/l) and glycerol (13.5 mg/l) as the main metabolites were obtained at 10 minutes treatment. Conclusion: It seems that ultrasound waves increased beta-carotene, and glycerol production of the cell by induction of defensive responses and secondary metabolism.

Abstract Aim: In this century, the production of recombinant drugs such as growth hormone has increased. Different problems existed in the expressions of cytoplasmic and Periplasmic types of growth hormone. Therefore, finding a way to optimize expression is very necessary. In this study, we optimized expression of growth hormone in the form of solution state by trx-tag method. This method increase protein expression (Periplasmic problem) and also prevents formation of inclusion body (problem cytoplasmic).
Material and methods: Gene synthesis and gene cassette was done in pET 32a expression vector. Gene cassette contains trx tag for protein solubilization, His tag for purification and enterokinase for separate rHgh from previous tags.
 Results: After cloning the gene in vector, its expression was confirmed by Western blot technique. The results showed that the expression of fusion protein was done well.
Conclusion: the obtained finding proved that protein can be soluble by Trx-tag and increased its expression levels. Moreover, better results can be achieved in the fermentation and downstream processing.

Abstract Aim: The main goal was to evaluate the impact of different incubation time intervals on human sperm DNA status using terminal deoxyribonucleotidyl transferase–mediated dUTP nick-end labeling (TUNEL) test.
Material and methods: This prospective study involved 21 normozoospermic specimens. After direct swim-up, sperm cells were incubated at 37^°C and DNA damage was evaluated at different time intervals (0, 1, 2 and 3 h).  After slide fixation with methanol 100% for 4 minutes and rinse in phosphate-buffered solution (PBS), TUNEL was added to each slide and incubated for 60 minute in dark. Eosin-Nigrosin and Papanicolaou staining protocols were applied in order to assess sperm viability and morphology, respectively.
Results: Sperm viability and normal morphology was improved after sperm processing (100%, 72.3% respectively, p < 0.0001). The rate of DNA damage was significantly higher after 2h compared to 0h (9.19±0.8% Vs 4.9±0.9%, respectively, p=0.008). Also there was significant difference in abnormal sperm DNA between 3h and 1h (10.95±0.7% Vs 7.1±1%, respectively, p=0.020).
Conclusion: Incubation of prepared normozoospermic samples at 37^°C more than 2 h may be associated with sperm DNA fragmentation. Therefore, it seems that incubation of human spermatozoa at 37^°C should be limited up to 2h prior to use in ART clinics. 

Abstract Aim: Recently, researchers have reported that BV (Bee Venom) has a potent anti inflammatory, anti tumor and anticancer effects. Besides, one of the essential aim in cancer therapy is restoring apoptosis. The Objective in this study was determination of apoptotic concentration induced by BV (Autumn, Semnan) on HL60 cancer cells.
Material & Methods: In this study, HL60 cells were purchased from the Pasteur Institute  in Tehran and were grown in RPMI-1640 medium supplemented with 10% FBS and 1% antibiotics (including penicillin and streptomycin) in the plate. After 2 hours, the cells were exposed by BV concentrations (2.5, 5, 7.5,12,15 µg/ml) for 24, 48, 72 hours. After passing desired time, the morphology of cells was determined under inverted microscope and cell viability was studied by MTT assay and determination of cell death was evaluated by flow cytometery assay and Hoescht staining.
Result: The morphological analysis and the results from Hoescht staining & flowcytometery Annexin V antibody exhibited that the cell death induced by BV was significantly apoptosis and bee venom by concentration of 12 µg/ml results in %50 of apoptosis cell death.
Conclusion: Our findings from this study showed that using lower dosage of BV during 48h treatment period cause inhibition of proliferation in time and dose dependent manner and higher dosage of 15 µg/ml cause cell lysis and necrosis. Experimental data showed that 12 µg/ml is inducer of apoptosis in HL60 cells.                                                                            
 

Abstract Aim: The aim was to investigate the interactions and cellular behavior of the blastema tissue and natural 3D elastic scaffold in vitro.
Material and Methods: In this study cows aorta was used as a scaffold. To prepare a very porous elastic scaffold, the cells and collagen were removed by treating that with 50 mg/ml cyanogen bromide in 70% formic acid. The prepared scaffold were then placed inside the blastmea rings and kept in culture media for 40 days. The interaction between blastema tissues and elastic scaffolds were studied in 10 days intervals.
Results: Microscopic studies in different day on blastema tissue and scaffold revealed that the cells and collagen fibers were omitted successfully from the elastic scaffold. Moreover, histological studies indicated that the cells had penetrated into the scaffold. In addition cell devision, probable differentiation of blastema cells to fibroblast and myocyte and also angiogenesis due to inductve effect of elastic scaffold were abserved.
Conclusion: Our results indicated that, it is possible to prepare a natural elastic scaffold from aorta by treatment with cyanogen bromide. On the other hand, this scaffold may have inductive effects on cell behaviors such as migration, adhesion, cell division and probably differentiation. Further studies are required to confirm the indentity of cells and other properties of the scaffold and also its possible use in engineering of vascular tissue.

Abstract Aim: The aims of study were to investigate callogenesis and regeneration of the studied cultivars and also exploring the possibility of gene transfer into maize by using complete bud and stem apical meristem explants.
Material and methods: Complete bud and stem apical meristem of six cultivars were used as explants on MS medium + 5 mg.L-¹ 2,4-D. For regeneration, calluses were cultured in MS including 1 mg.L-¹ Kn and 10 mg.L-¹ BAP. According to the results of tissue culture, SC703 and SC704 cultivars were selected for gene transfer. Transformation of explants was done with LBA4404 strain of Agrobacterium tumefaciens containing pBI121 vector. To investigate the transgenic nature of the samples, PCR and histochemical assay were used.
Results: Callus induction was taking place after 30 to 40 days and the frequency of callus induction in six cultivars was 100%, in addition the highest rate of regeneration (90 %) was observed in stem apical meristem explants of SC704. In some cultivars, stem apical meristem explants were more suitable, although in other cultivars, complete bud explants were better. PCR results for NPTII gene and GUS histochemical assay showed that some calli are transgenic and GUS gene is expressed.
Conclusion: The results of callogenesis, regeneration and genetic transformation showed that SC 704 was better than the others.
 

Abstract Aim: The aim of this essay was to assess the ointment effects of alcoholical extraction prepared from annual leaves of Verbascum speciocum grown in 'Bouein Zahra' area, on skin lesions made in Wistar rats.
Materials and Methods: In this study, at in vivo condition, 36 adult male Wistar rats were divided in 3 groups: Control (no treatment), Sham (treated with petroleum jelly) and Experimental (treated with ointment verbascum extract in different doses).
The operation day was considered as day zero. 10 days after treatment, up to the day of 14^th, after the surgery on the killed rats, the samples were collected from wound part of each rat for histological analysis and also collagen special staining and average of epidermis thickness, and hair follicle & blood vessels diameter were done.
Results: There was an increase in hair follicles’ diameter in experimental groups with 1/3LD50 Ointment comparing with control groups also a significant  increase was found in skin epidermis thickness in experimental groups with 1/4 LD50 versus control groups. Moreover a significant increase was observed in blood vessels’ diameter in experimental groups exposed to 1/3LD50 ointment to control.
Conclusion: In this study, we realized that the external use of V. speciocum extract on skin lesion with high dosage has a cytotoxic effects and were shown necrosis in the treated tissues but the lower dosages of extraction have been shown better wound healing.
 

Abstract Aim: Alpha smooth muscle actin (α-SMA) is an actin isoform present in many kinds of eukaryotic cells. In this survey the expression of this protein in skin appendages was examined.
Materials and Methods: In present study we examined the expression of this protein in 3 populations of dermal cells derived from three different skin appendages in vitro and in one case in situ by means of immunohistochemistry.
Observations: The results obtained here showed that in culture this protein is expressed nearly in all dermal papilla cells derived from vibrissa follicles. In case of dermal cells derived from claw unit a high percentage of cells expressed the protein, but in feather follicles only a small percentage of dermal papilla cells were positive for this protein. The α-SMA was also strongly expressed in dermal components reside at basal region of feather follicles in situ.
Discussion: Here we report for the first time that α-SMA is expressed in cultured dermal cells of claw and feather follicle. Moreover, these findings along with the results provided by other workers reinforce the crucial role of this protein in normal activities of skin and cutaneous appendages in mammals and other vertebrates. Given to a high expression of this protein in dermal cells of feather follicle particularly in situ, it can be concluded that α-SMA plays a more important role in birds compared to its role in hair follicle of mammals but the exact role of this protein needs to be elucidated. 

Abstract Aim: The aim of this study was to evaluate histometrical and stereological properties of the different tissue layers in male and female goats.
Material and Methods: To study the microscopic structure of the goat esophagus by stereological method, the eight healthy adult goats from both sexes were selected and the cervical region was dissected and esophagus removed. The obtained samples were fixed in 10% buffered formalin solution and then prepared tissue sections, stained with hematoxylin and eosin and studied using light microscopy.
Results: Results showed that there are significant variation in micrometry thicknesses of the submucosal and muscular layer at the cervical and thoracic regions (p < /em><0.05). The stereological results showed that mean volume of the esophageal muscular layer was significantly greater in male than female, but in other layers significant differences between both sexes was not observed (p < /em>>0.05).
Conclusion: Although morphometrical results showed that there are significant variations in different layers of the esophagus tissue, but stereological results showed no significant difference.
 

Abstract Aim: The somites are epithelial blocks of mesodermal cells differentiating to sclerotome and dermomyotome and notochord or axial mesoderm plays a major role in somitic cell differentiation to sclerotome. This study was aimed to evaluate the role of the notochord in sclerotomal differentiation of the somites in vitro. Material and Methods: In this experimental study, isolated notochords from chick embryo were encapsulated in alginate beads and co-cultured with somites for six days. In control group, the somites were cultured alone. Newly isolated and non-cultured somites were considered as somites on day 0. Eventually, morphology of cultured somitic cells was evaluated by inverted microscope and then RT-PCR method was used to analyze the sclerotomal and dermomyotomal gene expression profile. Results: In comparison with control group, the somitic cells co-cultured with notochord had highly differentiation potential and showed mesenchymal morphology with numerous slender processes. Gene expression profile of co-cultured somitic cells indicated upregulation of Pax1 and BMP4 and downregulation of MyoD, presenting sclerotomal cell characteristics. Conclusion: Embryonic notochord can induce in vitro sclerotomal differentiation in somites through upregulation of genes involving in this differentiation.

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