Cell and Tissue Journal

Abstract Aim: The role of Hsp70 chaperone from Rutilus frisii kutum in the thermal inactivation of luciferase in E. coli cell carrying Hsp70 and firefly luciferase was investigated.
Material and Methods: Co-transformation of E. coli cell was carried out with two expression vectors containing Hsp70 and firefly luciferase. The co-transformed cells carrying Hsp70 and luciferase were expressed under optimum conditions. After adding tetracycline, the cells were then incubated for 60 min at 40, 42, 44, 46, 48 and 50°C treatments. Finally, the luminescence activity of samples was calculated.
Results: After 30 min at 40 and 42°C temperatures, the luciferase activity in control samples reached almost zero, while in the co-transformed samples, about 72% and 60% of the luminance activity maintained compared with control samples (before heat treatment), and even after 60 min, up to 36% and 22% of the activity remained. Also, at 44 and 46°C temperatures in the initial times after stress, a significant difference was observed between the luciferase activity of the co-transformed and the control samples. In contrast, at 48 and 50°C temperatures, the changes of the luciferase activity of co-transformed samples were small compared with control samples even in the early stages of stress.
Conclusion: The thermal aggregation of luciferase as a significant reporter protein is inhibited at high temperatures via the activity of Hsp70 in the bacterial cell, which this process can be widely used in the food and pharmaceutical industries and the providing cancer diagnostic kits.
 

Abstract Aim: The aim of this study was to evaluate the biocompatibility and differentiation of human umbilical cord mesenchymal stem cells to osteoblast on prepared polycaprolactone scaffolds by electrospinning method, which is located under the surface modification by oxygen plasma.
Material and methods: After isolation of mesenchymal stem cells from human umbilical cord, flow cytometry analysis was performed. Biocompatibility of scaffold was examined using MTT assay. Cells morphology and their adhesion characteristics on the scaffold surface were studied using Scanning Electron Microscopic (SEM) images. Also biodegradability of scaffold calculated using weight loss method. Finally, for study the differentiation of cells on the scaffold surface, osteogenic differentiation medium was used and differentiation was performed by red alizarin staining and RT-PCR.
Results: Results showed that the cells not only had more suitable connection and reproduction abilities on the scaffold, but also were morphologically in a natural condition. Polycaprolacton scaffold is enjoyment of high biocompatibility and its biodegradability up to 15th day done with more rates in comparison with 15 next days. Also red alizarin staining and RT-PCR results showed high rate cell differentiation on polycaprolactone scaffold.
Conclusion: The obtained results of this research suggested that polycaprolactone scaffolds have potential usage as biocompatible biomaterials in tissue engineering and osteogenic differentiation applications.

Abstract Aim: The aim of this study is to evaluate the spermatogonial stem cells differentiation into male gametes.
Materia and Methods: Spermatogonial stem cells were cultured in T25 flasks after the enzymatically isolation of the testicular biopsies through azoospermic patients. In the third passage, cells divided into 4 different groups and were treated for 1 to 4weeks under the effect of a medium containing extracts of sheep testes as inducer and then expression of sperm maturation genes: Acrosin and Protamine1 were investigated by using of western blotting technique.
Results: After spermatogonial stem cells treatment by the extracts of sheep testes, variations were seen in cell shape and they convert into sperm- like. Moreover, Acrosin and Protamine1 expression were confirmed.
Conclusion: examination of induced cells showed that Acrosin and Protamin1 were expressed. Since Acrosin and Protamine1 are the major proteins of the spermiogenesis, it could be concluded that these cells had been completed spermatogenesis stage and started the spermiogenesis stage.
 

Abstract Aim: In this study zinc oxide nanoparticles (ZnO nanoparticles) toxicity effects on some male sexual hormone levels in blood serum and testicular tissue in adult intact rats was studied.
Material and Methods: The rats were divided into a control and four treatment groups. ZnO nanoparticles at concentrations of 2.5 and 1 g/kg were orally administered for two treated groups and intra-peritoneal were synchronic used for two other treated groups. Nanoparticles effect and its damages on testicular tissue were studied one, seven and fourteen days after treatment.
Results: Blood serum tests showed that serum LH and FSH levels increased in zinc oxide nanoparticles treated groups in comparison with control (p < /em><0.05). Also, testes tissue slides examination showed specific tissue damages in all four zinc oxide nanoparticles treated groups in comparing to control. These injuries represent irregularities in the cell layers, cell-free seminiferous tubules, congestion in seminiferous tubules area and changes in spermatogene cell nucleus.
Conclusion: Results of the present study show nanoparticle zinc oxide effect on morphological structure and sex hormones.
 

Abstract Aim: Plants face many stresses in nature and have different defense mechanisms against stress. Rice is a cold sensitive plant and its product considerably decreases under cold stress. Since rice is an important crop, recognizing the tolerant cultivar and their defense mechanism against low temperature seems necessary.
Materials and Methods: In the present study anatomic and morphological features of tolerant (Onda) and sensitive (Nemat) rice varieties were studied at the germination phase under low (10, 15^°C) and room temperature (25^°C) for a period of of 8 days.
Results: Results show that dry weight, length of the hypocotyl and radicle of both Onda and Nemat cultivars under stress were decreased. The decrease was greater in the Nemat cultivar than the Onda cultivar. In addition, anatomic features of both cultivars under cold stress underwent some changes including increase of thickness of suberin layers and decrease of the amount of aeranchyma tissues.
Conclusion: Changes observed in the two varieties of rice were towards increasing resistance to low temperature stress. The Onda variety had a more suitable defense mechanism compared to the Nemat variety.
 
 

Abstract Aim: Movement of water across cellular membranes is facilitated by the presence of water channels named aquaporins. Plant plasma membrane intrinsic proteins (PIPs) fall into two groups, PIP1s and PIP2s that exhibit different water channel activities. So PIP1s are inactive, whereas PIP2s induce a marked increase in the membrane osmotic water permeability coefficient, p < /em>_f. This difference may be due to their subcellular localization. The aim of this research was to determine the subcellular localization of NtPIP1;1 and NtPIP2;1 in living tobacco cells.
Material and methods: In this experimental study, NtPIPs cDNA sequences were fused to green fluorescent protein gene and expressed transiently in tobacco mesophyll protoplasts. Then, cellular localization of aquaporins was investigated using fluorescent microscope under blue excitation wavelength.
Results: When expressed alone, NtPIP1;1 fusion protein was retained in the internal membrane structures, whereas NtPIP2;1 was found in membrane structure within the cell. To identify the internal structures containing NtPIP1;1 fusion proteins, NtPIP1;1-GFP was co-expressed in tobacco protoplasts with the YFP::HDEL protein, an ER marker. As shown, NtPIP1;1-GFP and YFP::HDEL colocalized.
Conclusion: NtPIP2;1 was transported to plasma membrane, but NtPIP1 was retained in the ER of tobacco protoplasts so showed no water transport activity.
 

Abstract Aim: The present study aims to investigate the effect of simultaneous encapsulation of DNA with different ratio of PLL via PLA-PEG copolymer, on gene delivery efficiency into mammalian cells. Some characteristics such as biocompatibility, DNA protecting against restriction enzymes, DNA release rate, size and zeta potential were also investigated.
Material and Methods: PLA-PEG/PLL/DNA nanoparticles with different ratio of PLL were prepared by double emulsion-solvent evaporation technique. Then, the release percentage of DNA and the zeta potential of particles in Phosphate Saline buffer (PH=7) were measured.
Results: Increasing the PLL percentage in the PLA-PEG/PLL/DNA nanoparticles resulted in enhancement of particles size and zeta potential. Gel electrophoresis analysis of DNA extracted from PLA-PEG/DNA and PLA-PEG/PLL/DNA nanoparticles after treatment with DNase I enzyme indicates the ability of PLA-PEG and PLA-PEG/PLL copolymers in DNA protection. The flow cytometry and fluorescent microscopy study confirmed that gene transfer efficiency was improved by increasing the ratio of PLL in PLA-PEG/PLL/DNA nanoparticles.  Moreover we found that gene transfer efficiency was one and half fold higher then PLL/DNA complex in the serum medium.
Conclusion: The results showed that PLA-PEG nanoparticles in the serum medium have higher potential gene delivery to the MCF-7 cells in comparison with PLA-PEG/PLL/DNA nanoparticles.
 
 

Abstract Aim: In the present study, we examined the possibility of astaxanthin usage in the MS model of mice, experimental autoimmune encephalomyelitis (EAE).
Material and methods: EAE disease was induced using MOG35-55 peptide and complete Freund adjuvant (CFU) in female C57BL/6 mice. Mice were divided into control, patient group (MS model, EAE), and prevention group, the group received astaxanthin for 21days before immunization, and astaxanthin treatment was initiated when mice showed the first symptoms of EAE. Each group consisted of 8 mice. The consumption of Astraxanthin was continued for 35 days, and each mouse received astaxanthin about 0.4% of the weight of used pellet (about 400 mg). Cell proliferation was measured by MTT, IL1β, but IL6 levels (proinflammatory cytokines) were measured by ELISA. In addition, the spinal cord tissues were also taken for immunohistopathological evaluations.
Results: Based on the results, the use of astaxanthin produced appropriate clinical and pathological outcomes, lead to decrement of IL1β, and IL6 proinflammatory cytokines (p < 0.0001) production. Immunohistopathologic studies also showed a decrease in leukocyte infiltration in the spinal cord of experimental groups.
Conclusion: Overall, this drug approach can be considered as a useful strategy in the prevention and treatment of multiple sclerosis.
 

Abstract Aim: This study was carried out to evaluate association of ISSR and RAPD markers with fruit traits of hazelnut (Corylus avellana). Material and Methods: In this research, associations between ISSR and RAPD markers with fruit traits were investigated in 35 genotypes of hazelnut by applying multiple regression analysis (MRA). Results: Some ISSR and RAPD fragments were found to be associated with seven of affecting fruit traits. Some of ISSR and RAPD markers were associated with more than one fruit trait in multiple regression analysis. Such an association may arise due to pleiotropic effect of the linked quantitative trait locus (QTL) on different traits. For example, some informative markers were associated with both nut length and kernel weight traits. Also, some informative markers showed association with all nut length and weight and kernel weight. Also, some markers had correlation with both kernel length and weight, indicating a significant correlation among these traits. Conclusion: It is possible to use these markers along with morphological traits in hazelnut breeding programs for identification of suitable parents to produce mapping populations and hybrid cultivars. Also, these results could be useful in marker-assisted breeding programs when no other genetic information is available.

Abstract Aim: In the present investigation, the effects of grape seed (Vitis Vinifera) alcoholic extract were examined on skin Histomorphometric wound healing changes in diabetic Wistar rats.
Material and methods: forty eight male wistar rats were divided into four groups: negative control, positive control, the first experimental and the second experimental. In all of treated rats, a scar in the length of three centimeter was created on the left of the vertebral column. The wound healing was evaluated microscopic as well as macroscopic. The methods Mann – Whitney, Kruskal – Wallis were used for data analyses. The used software was SPSS ver. 16. 
Results: agglutination in diabetic groups was longer than normal, while the process in the group treated with V. vinifera was quicker than the control and also first experimental groups, respectively.
Conclusions: the obtained results showed that V. vinifera ointment accelerate the wound healing in normal and diabetic samples.

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: The purpose of this study was to investigate the effect of adding different levels of zinc (Zn) to semen extender on the sperm parameters after freezing and thawing in rams.
Material and methods: The spermatozoa of five Farahani rams breed (with a mean age of 3 ± 0.5 years and weight of 60 ± 5 kg) were taken twice a week. The semen was collected from the rams by artificial vagina and then the samples were mixed to removing the individualistic effects. The sperm samples (in five replicates) were randomly divided into four groups (zero, 50, 100 and 150 μM of zinc sulfate). Each falcon had the desired diluent that was mixed with Zn and tris and prepared for freezing steps. Samples were stored in semi-milliliter straw and kept with samples under the same conditions and temperatures of 37 °C. Moreover, in these conditions were filled and frozen. Sperm samples were evaluated after thawing for motility, progressive motility, survival (eosin-neogrosin test), sperm membrane integrity (HOST) and sperm morphology (Hancock test).
Results: Results showed no significant difference in the motility percentage and sperm membrane integrity in zinc sulfate environments compared with the control group. The treatment with 50 μm of zinc sulfate showed a higher level of significant (P˂0.05) in the percentage of progressive motility compared with the control group. In sperm viability test, the treatment with 100 μm of zinc sulfate was significantly different from those of 50 and 150 μm zinc sulfate treatment (P˂0.05).
Conclusion: application of zinc sulfate in ram semen extender might be improving some of sperm parameters after the freezing-thawing.
 

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.