مجله سلول و بافت
سال پنجم شماره 2 (تابستان 1393)

Petroleuciscus esfahani، is an important fish species which shows very wide distribution in the Zayandehrood River. As there was no scientific report regarding hematological characteristics of this species، therefore in this study، some major hematological indices of this fish from different stations of the river were analyzed. Totally، 127 fish were caught from 4 stations along the river including Cheshmeh dimeh، Khersonak، Chamgordan and Pol-e-Safaiyeh and blood was collected from their caudal vein using heparinised syringe. Then some routine hematological indices were measured using standard methods. The mean red blood cell counts، hematocrite and hemoglobin were in the range of 3. 02-3. 4×106 cell. mm-3، 28-34. 68 % and 9. 14-11. 69 g. dl-1 respectively without any significant differences among stations (P>0. 05). However، a significant elevation in number of white blood cell (WBC) counts in Chamgordan and Pol-e-Safaiyeh (down-stream) as well as significant changes in differential WBC (DWBC) counts and secondary hematological indices specially in Chamgordan station in comparison to other station were observed (P<0. 05). The differences in some blood characteristics such as WBC and DWBC may be reflecting the existence of some environmental pollution such as heavy metals in Chamgordan station.

Different methods have been used for introducing DNA into mammalian cells. Calcium phosphate-based transfection protocols are inexpensive and easy to perform، however، the transfection efficiency in some cases is not satisfactory. In this study we try to increase the efficiency of calcium phosphate-mediated cell transfection. 2. 0×105 HEK293T cells were plated in each well of a 6-well plate containing a 10% poly L lysine-coated coverslip and grown to 60% confluency. The cells were then transfected with a plasmid encoding GFP (Green Fluorescence Protein)، using a standard calcium phosphate protocol in the presence or absence of chloroquine phosphate. GFP protein expression was observed by immunofluorescence microscopy and quantified with the help of Image J software. We have shown that chloroquine phosphate can highly improve the efficiency of calcium phosphate-dependent transfection in the HEK293T cells، leading to an approximate 8-fold increase in the number of transfected cells. Chloroquine phosphate was not toxic to the cells، however، its positive effect on transfection efficiency was abolished in the presence of glycerol or DMSO (Dimethyl sulphoxide). we recommend chloroquine phosphate for increasing the efficiency of transfection by the inexpensive method of calcium phosphate. In addition، we recommend that chloroquine phosphate to be used alone and not along with other inducers of cell transfection.

The aim of the present study was to investigate the effects of ASA (acetyl salicylic acid) on pyramidal neurons of the hippocampal CA1 sector and spatial learning following transient middle cerebral artery occlusion (MCAO) in rats. Forty-nine male rat were randomly divided into control، sham، vehicle، ischemia and ASA (20، 40 and 80 mg/kg) groups. Transient focal cerebral ischemia was induced in rats by 20 min of MCAO. Animals received ASA or Vehicle by i. p 30 min after stroke onset. Four days after ischemia، animals were subjected to 4 days of training in the Morris water maze (MWM) with the invisible platform to test spatial learning. At the end of behavioral test، rats were sacrificed and pyramidal neurons in the CA1 sector were stained with Hematoxylin and Eosin (H&E) and observed by light microscope. MWM showed that the MCAO caused disturbance of spatial learning in rats، and ASA treatment could not improve the spatial learning function. But، ASA treatment significantly reduced the number of degenerating neurons in hippocampal CA1 region compared with vehicle group (P<0. 01). These results suggest that ASA injection، 30 min after stroke onset in rat decreases neuronal injuries following ischemia.

Recently، attention has been diverted to the pluripotency characteristic of Spermatogonia Stem Cells (SSCs) in in-vitro culture. The aim of this study was to evaluate the morphological and molecular genetics changes of differentiated SSCs to Embryonic-like Stem cell (ESC). The SSCs were collected from neonatal mouse testis via a two-step mechanical and enzymatic digestion and cultured in DMED containing 15% FBS. ES-like cells colonies was appeared in the fifth passage. The expression of pluripotency and spermatogonia specific genes; Nanog، SOX2، Klf4، Stra8، DAZL، Plzf as well as the expression of puripotency markers Oct4 and Alkaline Phosphatase Activity (ALP) of ESC respectively was investigated using RT-PCR and immunocytochemical techniques and compared with ESCs. The pluripotent characteristic of ES-like cells derived from SSCs was confirmed based on morphological investigation and high expression of ALP as well as pluripotency marker Oct4. The expression of specific spermatogonia genes like Stra8، DAZL، Plzf was decreased and the expression of pluripotency genes such as Nanog، SOX2، Klf4 was increased during the differentiation of SSCs to ES-like cells. In vitro culture of SSCs was conformed to induced ES-like cells. This process was accompanied by wide alteration in molecular profile expression of specific spermatogonia and pluripotency genes. In over all، SSCs can be considered as an opening window to generate pluripotent stem cell.

Myostatin gene or growth and differentiation factor 8 (GDF8) which has been identified as the factor causing muscle doubling، undergoes a series of mutations that causes gene inactivation. In bovine the loss of gene activity has been associated with double- muscled phenotype.. Due to the role of myostatin gene in muscle development، the objective of this study was to analyze the coding region containing mutations which potentially altering the myostatin gene expression in Farahani sheep. DNA from blood samples of eighty six Farahani sheep was extracted and used to amplify a 337-bp fragment in myostatin gene. Restriction fragment length polymorphism (RFLP) of the PCR product was performed by addition of enzyme and then PCR-RFLP genotypes were analyzed. Genotype frequencies of MM، Mm and mm were characterized as 0. 046، 0. 127 and 0. 827، respectively. In addition the allelic frequencies for alleles M and m was estimated as 0. 11 and 0. 89 respectively. The results of this study indicated that the Farahani sheep was polymorphic for myostatin، but this population was not at Hardy- Weinberg equilibrium.

Manipulation of cell culture media by elicitors is one of the important strategies for inducing secondary metabolism production as valuable metabolites. Salicylic acid (SA) as a biotic elicitor is effective for increasing medicinal metabolites. The aim of this study was to evaluate the effectiveness of salicylic acid on the growth، some of physiological parameters and trigonelline production in fenugreek cell culture. The cell was obtained from hypocotyl explants of fenugreek on MS solid medium supplemented with naphthaleneacetic acid (0. 4 mg/L) and benzyladenine (0. 4 mg/L). Salicylic acid with concentrations of 12. 5 and 25 and 50 mg/L were used. After one week، some of physiological parameters and trigonelline production in fenugreek cell culture were investigated. Following treatments with salicylic acid، the results showed that the cell growth and viability of cells were decreased as compared to control. The treatments increased hydrogen peroxide content and lipid peroxidation rate in cells compared to control. In addition trigonelline and total phenol production were improved by all the treatments. Treatment of cells with 50 mg/L salicylic acid increased trigonellin 2- fold higher than control. Salicilyc acid can be used as an elicitor in fenugreek cell culture and induce more trigonelline production.

The effects of different concentrations of ZnO nanoparticles on some growth parameters، the amounts of proline and alkaloids and changes in activity of some antioxidant enzymes were investigated. Experiments were performed under controled greenhouse conditions and completely randomized design with three replications. Plants were exposed to different concentrations (2، 4، 5، 10 and 15 mM) of nano zinc oxideand the effect of these concentrations were compared with the plants that used a complete nutrient solution containing zinc Sulfate. Treatment of the plant with concentration of 0 to 2 micromolar caused increased and higher concentrations significantly reduced dry weight as well as root length. The results showed that the antioxidant enzymes activities in the treated plants increased with increasing concentration of nano zinc oxidein the nutrient solution compared to the control plants. As the level of stress was elevated، the amount of proline and total alkaloids also increased. With increasing concentrations of zinc oxide nanoparticles، Zn absorptionraised. Due to this toxicity، plant growth parameters were affected and oxidative stress in plants was induced. To overcome free radical production، plants have increased their antioxidant enzyme activities and proline.

The contamination of soil by crude oil is an important environmental problem. Bioremediation have gained more attention in recent years as a low-cost technique for pollutant removal. The aim of this research was to investigate the ability of rhizospheral fungal strains collected from plants grown in a petroleum-polluted soil using different concentrations of pollutant. In this research rhizospheral fungal strains were collected from the plants grown in the Tehran refinery area and cultured on the PDA media containing different concentrations (0-15%) of crude oil. Efficiency of removal for each isolated fungi was determined after one month. Results showed that، all fungi were able to remove petroleum at the concentrations of 1، 2، 4 and 6% (P≤0. 01). In addition all the fungi except Diplosporium sp. and Penicillium sp.، were able to decrease crude oil at the concentrations of 8، 10 and 15% (P≤0. 05). It was concluded that the studied fungi were able to remove the crude oil from the growth media. Since Aspergillus sp. was relatively more effective therefore this fungus has the highest bioremediation potency of petroleum polluted environments.

The aim of this study was to investigate the effect of cold temperature and cold water stresses on chlorophyll fluorescence parameters in different rice genotypes. Six rice genotypes including Koohsar، Shiroodi and Tarom Hashemi cultivars as well as PSB44، RI96 and IR752 lines were grown under hydroponic conditions. After two weeks، plants were kept as control (28 °C)، cold temperature stress (8 °C) and cold water stress (8 °C) then chlorophyll fluorescence parameters were measured. Cold temperature stress significantly reduced some of the measured parameters including variable fluorescence (Fv)، maximum photochemical quantum yield of photosystem II (Fv/Fm)، effective photochemical quantum yield of photosystem II [Y (II)]، photochemical quenching coefficient (qP) and electron transport rate (ETR); while significantly increased quantum yield of regulated non-photochemical [Y (NPQ)]، quantum yield of non-regulated non-photochemical[Y (NO)]، non-photochemical quenching (NPQ) and Non-photochemical quenching coefficient (qN). Whereas cold water stress had significantly affected minimum fluorescence (Fo)، maximum fluorescence (Fm)، Fv and ETR. Inaddition PB44 line and Tarom Hashemi cultivar in cold temperature stress and RI752 and PB44 lines in cold water stress had the minimum change in fluorescence parameters. cold temperature stress was more effective than cold water stress. In cold temperature stress، PB44 and Tarom Hashemi were most tolareant genotypes while RI96، IR752 and Shiroodi were the most susceptible genotypes. In cold water condition، RI752 and PB44 were the most tolerant genotypes while IR96 and Shiroodi genotypes were the most susceptible genotypes.

the aim of this study was to investigate the effect of iron on cell division، interacellular beta-carotene and chlorophyll synthesis in unicellular green alga Dunaliella. In this study 4، 32، 64، 128، 256 and 512µM concentrations of iron was applied on D. bardawil and D. salina. The culture media were kept in a growth chamber in a light/ dark regime of 16/8 hours at 26±2 °C. Then beta-carotene، chlorophyll content and cell divisions were determined. In both species the maximum cell growth was observed in 4 µM and the amount of chlorophyll as well as cell growth decreased by the increase of iron concentration in the medium. The maximum reduction of the chlorophyll and cell growth was observed in 512 µM، where as the maximum intracellular beta-carotene content was shown in 128 µM. Results showed that probably free radicals contents of both species have been increased in presence of iron. Iron treatment decreased the chlorophyll content to reduce the free radicals produced due to excitation of chlorophyll molecules. Moreover، the cells produced high amount of beta-carotene as an antioxidant against free radicals in presence of Iron. It seems D. salina has better performance in presence of high concentration of iron than D. bardawil due to less synthesis of beta-carotene and more production of other antioxidants as well as reduction of chlorophyll production.