JOURNAL OF CELL & TISSUE
Year:2016 | Volume:7 | Issue:2 |Papers Count:10

Aims: The aim of this study is healthy and desirable plants production of two Crataegus species using meristem culture and somatic embryogenesis and synthetic seed preparation that is a biotechnological result.Material and methods: Strile seeds, vegetative - generative organs pieces and the buds of two Crataegusspecies (Crataegus pseudoheterophylla and C. microphylla) were cultured on solid MS medium with auxin hormones (IAA, NAA and 2, 4- D), cytokinins (KIN, BAP, Zeatin) and GA3 under sterile conditions. Samples were capsulated by sodium aglinate and calcium chloride.Results: Leaves, apical meristems and axillary bud meristems explants were the best for callus production in solid MS medium with some concentration of auxins and cytokinins. Embryoids and especially embryos were almost obtained from leaf explants or sub culturing of embryogenic callus in solid MS medium contains auxins, cytokinins and a little of NaCl. Early autumn and late winter axillary buds were suitable for synthetic seed production. Sodium alginate %3 and calcium chloride %1 were used as a capsule for synthetic seed production. For artificial seed storage 4°C and for germination of these seeds 24 ± 2oC were suitable.Conclusion: There are some difficulties for somatic embryogenesis in two studied Crataegus.Hormones type and dosage are very important and effective on callus, embryoids and embryos production. Buds harvesting time is also important for synthetic seed production. The supplements of buds with MS medium and capsulation those with sodium alginate are favorable.

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

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

Aim: The aim of this study was to convert lamp and solar energy into electrical energy with bacteriorhodopsin protein monolayer, evaluate examined compounds (TiO2, FTO, Br) to increase the biological photocells efficiency, photocells building feasibility for use in architecture, military and biological photocells production.Material and Methods: Titanium dioxide nano-particles were placed on the surface of a fluorine tin oxide (FTO) using Belding method. Bacteriorhodopsin (Br) monolayer was prepared by Langmuir-Blodgett (LB) technique and photo anodes were made. Then the FTO glass was covered with an H2PtCl6 solution drop (2 mg Pt in 1 ml EtOH), and cathode was prepared. Photo anodes electrode (FTO/TiO2/Br) and opposite electrode (cathode) (FTO/Pt) were placed as a sandwich.Results: Light dependent pH changes, demonstrated bacteriorhodopsin activity. The created monolayer photocells power was 0.7mA and their voltage was 190 mV, which was different from the accumulated photocells powers (0.5 mA), 0.2μA with its voltage (165 mV), 25 mV, indicating that monolayer cells had a better accumulated efficiency.Conclusions: Langmuir technique is a very suitable method for bacteriorhodopsin protein stabilization as a monolayer, on the nanoparticles substrate (titanium dioxide) with maximum activity maintenance.

Aim: Leukemia inhibitory factor (LIF) has beneficial effects on neural stem cell function and neurodegenerative disease treatment. However limited data is available about the effects of LIF overexpression in neural stem cells on brain tissue changes. In this study transgenic rats (TR) with over expression of LIF under the control of nestin gene promoter were produced and histological changes of their hippocampus were studied.Material and methods: To produce transgenic rats, promoter regions of pIRES2-EGFP vector were replaced with 2nd intron nestin promoter and Hsp68 enhancer sequence, and injected into testes (10mg) of 10 male rats. After mating with female rats, trangenesis confirmation was done using conventional PCR, real time PCR and fluorescent microscopy. Brain of mature transgenic and control rats (4 months of age) were obtained, sectioned and stained with H & E and then their hippocampus structure were studied.Results: LIF gene was significantly expressed in higher levels in hippocampus stem cells of TR comparing to control (P<0.05). This finding in concomitant with presence of EGFP signal in neural stem cells of hippocampus confirmed the transgenesis in offspring. Oligodendrocytes number was significantly higher in hippocampus of transgenic rats in comparison with control groups (P<0.05).Conclusion: Over expression of LIF in neural stem cells resulted in elevation of oligodendrocytes in hippocampus of rat brain. Because oligodendrocytes destruction are importance in neurodegenerative diseases progression, it is possible that successful LIF transmission into the brain can be used in the future to restore the oligodendrocyte mass.

Aim: The aim of this study was to determine whether varicocele can cause any changes in number and size of ovarian follicles.Materials and methods: In this study, 30 adult female Wistar albino rats in 3 groups (n=10) consisting of: 1. control, 2. Sham and 3. varicocele are prepared. Varicocele group were operated and left ovarian vein was respectively closed. After 2 months the animal scarified and the ovaries taken for histological process. Statistical analysis was performed using the one way ANOVA and Turkey’s tests were used forpost hoc multiple comparisons, was considered statistically significant.Results: There is no significant differences in rat and ovary weights before and after varicose induction between groups. The number of primordial follicles in varicocele group increased significantly. The number of primary, secondary and graffian follicles in varicocele group decreased significantly. The size of secondary follicles in varicocele group decreased significantly (P<0.05).Conclusions: Finally the results of this study shows that varicocele may lead to increased female infertility through various factors, which includes changes in follicular growth.

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 RTPCR.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.

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 (χ2=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.

Aim: In this study gill and mantle histopathological and ultrastructural changes of freshwater bivalve, Anodonta cygneafrom Unionidae in exposure to sub-lethal concentrations of copper oxide nanoparticles for 14 days were investigated.Materials and Methods: Forty numbers of A. cygnea (length: 11.5 ± 0.9 cm; weight: 84.5±6 g) were collected from Tajan River estuary, Sari, Mazandaran Province and after transferring to the laboratory were divided into two control and treated groups. Treated group were exposed to 25 ppm CuO nanoparticles for 14 days. Gill and mantle tissue sections of control and treated samples were taken in days 4, 9 and 14, and then prepared for light microscopy using hematoxylin and eosin (H & E) staining method. Samples were studied by scanning electron microscopy (SEM) after fixation, drying and preparation.Results: Light microscopy results showed significant histological alterations in all of studied organs of exposed bivalves compared to the controls. Changes were hyperplasia, hypoplasia, gill filaments shape and dimension changes and also haemolymph channels adema. Also increasing mucous cells number and external epithelium hyperplasia in mantle were observed. SEM analyses showed adverse destruction of gill filaments and blocking water channels in gill specimens, as well as, mantel necrosis and indentation.Conclusion: according to obtained results, it can be concluded that short time exposure to nanoparticles may lead to noticeable changes evident in histological and ultrastructural internal organs of freshwater bivalveAnodonta cygnea.

Aims: Given the importance of the identification of scientific and commercial communities of Artemiaspecies, especially hybrid populations, some phenotypic and genotypic characteristics of hybrids between A. sinica and A. urmiana were studied in vitro.Materials and Methods: breeding two species of brine shrimp from hatching to maturity in standard laboratory conditions was provided. Then 64 males and females of each species were separated and mutual hybrid production was done in 50 ml tubes. Daily, the larvae were separated and reared partly and independently. Studies of fatty acids profile and survey by restriction enzyme pattern of 12S-16S mtDNA genome withHpaII enzyme in comparing of the parents and offspring’s of first-generation were done.Results: Comparing hybrids fatty acids profiles to pure parents showed that some fatty acids amounts are highly dependent on parental origin. In some samples the same restriction enzyme pattern was observed in male parents and the first generation hybrids that was unexpected.Conclusion: The results revealed that fatty acids profile is strongly influenced by maternal or paternal origin genes of inheritance. Therefore, by directional selection of parents it is possible to produce special Artemia offspring’s with unique phenotypic characteristics. Comparing of enzymatic digestion patterns with the first generated hybrid revealed that the mitochondrial genome (or mitochondria) also is inherited from the sire.