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Objective: Dendritic cells have a critical role in control and regulation of immune responses. It is believed that these cells can be used for the treatment of many diseases. One of the methods used in immunotherapy is based on generating of tolerogenic dendritic cells through inhibition of expression costimulatory molecules. CD40 is one of the costimulatory molecules, and inhibition of expression by antisense or siRNA techniques, can generate tolerogenic dendritic cells. Generation of tolerogenic dendritic cells will be useful in the treatment of many diseases. By developing a quantitive RT-PCR for evaluation of gene expression, generation of these cells could be possible. Using proper software we designed an Antisense and transfection of dendritic cells by lipofectamine 2000 (Invitrogen) could lead us to generate tolerogenic dendritic cells. Materials and Methods: In this study dendritic cells were extracted from of Balb/c mice Spleen and the purity of this extraction was determined by flow cytometry. BCL1 cell line as a CD40 expressing control group and Wehi-164 cell line were cultured in RPMI-1640+10%FCS. Primer design for CD40 gene and house keeping gene (GADPH) was done by bioinformatic soft wares such as Beacon designer, mfold and Blast. RNasy plus mini kit (Qiagen) was used for RNA extraction and the Purity and integrity were determined by O.D at 260/280 and agarose gel electrophoresis. In the next step cDNA synthesized and quantitative RT-PCR for CD40 using IQ sybergreen (Biorad) were setup. Finally, standard curve for CD40 and internal control in different RNA concentrations were performed. After transfection with lipofectamin 2000 the amount of gene suppression were quantified by qualitative RT-PCR. Results: Using gradient real time PCR, optimum annealing temperature, Ct and ∆Rn for CD40 and GADPH were determined, annealing temperature was 59.5ºc and melting temperature was 84°c. Slope of the curve and the efficacy of PCR for CD40 and GADPH genes were quantified by serial dilution method

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Background and Aim: Toxins produced by Clostridium botulinum are among the deadliest compounds known that cause botulism. Currently, the detection of BoNTs in food using bioassays on laboratory mice is a very sensitive method with a detection range of 7 to 20 pg.mL^-1. However, bioassay for mice is time consuming. This method is fast, highly specific, and sensitive to experiments on mice. The aim of this study was to use the modified Sandwich ELISA method to detect BoNT/B toxin.

Materials and Methods: Recombinant 370 amino acid protein was expressed from the carboxyl terminus of the binding moiety of BoNT / B toxin with a molecular weight of 45 kDa as antigen and purified by Ni-NTA affinity chromatography. IgG antibodies were isolated from mouse and rabbit sera byG protein affinity chromatography. The sensitivity and specificity of the method designed to detect recombinant BoNT/B-HcC antigen and botulinum toxin type B were evaluated.

Results: Purified rat and rabbit antibody concentrations were 3 and 4.5 mg / ml serum, respectively. The minimum concentrations of detectable protein were determined by indirect ELISA with purified mouse and rabbit antibodies at 475 and 118 pg. By optimizing the sandwich ELISA method, at least 30 ng of recombinant BoNT/B-HcC antigen and146 pg of highly specific BoNT/B were detected.

Conclusion: sandwich ELISA method can be used for accurate and sensitive identification of Clostridium botulinum toxin type B. It is necessary to evaluate the effectiveness of this method in the future to detect botulinum toxin in environmental and food samples.
 

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The short–term inflation dynamics and its cyclical interactions with real economic variables are basic issues in the context of monetary policies analysis. This study investigates and estimates the hybrid new Keynesian Phillips curve for Economy of Iran during 1971-2008. On this curve, the effective variables on current inflation would be future inflation, lagged inflation and GDP gap. This paper makes use of three Kalman, Hodrick- Prescott and band-pass filters to estimate GDP gap. There is a structural break in 1979 due to victory of the Islamic Revolution in Iran. Findings indicate that GDP gap has a significant positive impact on current inflation, which means the effectiveness of real variables, besides monetary policies, on inflation in the long-run. Our findings are consistent with other Phillips curve models, which confirm the effective role of output gap on current inflation. In addition, the coefficients of the expected inflation and lagged inflation variables are statistically significant, which indicate that firms look forward and backward in setting prices, but coefficient of expected inflation variable is higher than that of lagged inflation, means that firms pay more attention to the expected inflation in setting current prices. The evaluation tests indicate the accuracy and reliability of models.  

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     The objective of this study was to extract the essential oil of purple basil leaf, to identify its compounds and to investigate its antimicrobial effects on some food-borne pathogenic bacteria through different qualitative and quantitative methods, and eventually, to compare it with some antibiotics including vancomycin and gentamycin in vitro. The essential oil components were identified with GC/MS. The antimicrobial effect of basil essential oil was measured through well diffusion agar (WDA), and finally, the minimum inhibitory concentration (MIC) of the essential oil was determined using microdilution broth and triphenyl tetrazolium chloride. The results revealed that 28 identified compounds constituted 99.28% of the whole essential oil compounds. p-Allylanisole (51.64%) was the most abundant component of the essential oil. In addition, other main components such as n-Tricosane (24.83%) and Linalool (14.81%). In the wall in agar method, the mean free zone diameter was equal to 15.9 mm in the case of Gram-positive bacteria and 11.15 mm in the case of Gram-negative ones. The minimum MIC of purple basil essential oil ranged from 4.6 to 36.8 mg/ml in the case of pathogenic bacteria. Moreover, its minimum bactericidal concentration varied from 4.6 to 73.6. in conclusion, it can be said that purple basil essential oil was effective on Gram-positive bacteria at lower concentrations and could inhibit their growth.

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The aim of this study was to investigate the in vitro proliferation of Astragalus adscendens. Explants were taken from hypocotyl and cotyledon and were cultured on the basic medium of Murashige and Skoog (MS) complemented with various plant growth regulators, (NAA, BAP, KIN, ZEA), to induce direct shoot regeneration. Callus induction was significantly affected by different concentrations of PGRs. Callus formation was observed from hypocotyl explants, but they were not induced to adventitious shoot regeneration and most of them were turned into brown. Therefore, rapid multiplication, performed using shoot apical buds, and obtained from 15-day old sterile seedlings. Apical buds were cultured on MS medium containing various levels of BAP, KIN and ZEA (0.5, 1.0, 2.0 and 4.0 mg L^-1) alone or in combination with 0.5 mg L^-1 NAA. The highest number of shoot regenerants (8.5/explants) and leaves (22.4/explants) obtained on MS medium with 4 mg L^-1 BAP. The highest root induction (100%) was obtained from MS media supplemented with 0.3 mg L^-1 NAA. Rooted plantlets were successfully acclimatized in pots with 1:1:1 mixture of soil, peat, and perlite.

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In most semiarid and arid areas, fresh water shortage compels managers to use low quality water sources with high salinity to irrigate turf and landscape. Recent research has noticed that management of nitrogen fertilization can alleviate salinity effects on plants. This greenhouse sand culture experiment was conducted in order to investigate morphological and physiological responses to salinity stress in Kentucky bluegrass (Poa pratensis L.) grown using different nitrogen sources. Three salinity levels (0, 40 and 80 mM NaCl) and three NO₃^-/NH₄⁺ ratios (6/0.5, 6/1 and 6/2) were applied in nutrient solutions. Under non saline conditions, higher ammonium concentration increased Turf Quality (TQ), leaf NO₃^-, proline content, Nitrate Reductase Activity (NRA), shoot and root growth. On the other hand, leaf potassium (K⁺) sodium (Na⁺) and MalonDiAldehyde (MDA) content were not affected. During the first week, the 40 mM NaCl treatment showed that the positive effects of NH₄⁺ on salinity tolerance were still perceptible. However, the 80 mM NaCl treatment showed that the adverse effects of high salinities were more pronounced when turf received high ammonium rate nutrient solution, as manifested by the decrease of TQ, NO₃^-, NRA, K⁺/Na⁺ ratio, shoot and root growth and by the increase of leaf MDA content. This suggests that effects of NO₃^-/NH₄⁺ ratio on salt tolerance varies with salinity levels.