Inotropic and chronotropic effects of 6-hydroxy-4-methylquinolin-2(1H)-one derivatives in isolated rat atria

Background: Selective phosphodiesterase (PDE3) inhibitors improve cardiac contractility and may use in congestive heart failure. However, their proarrhythmic potential is the most important side effect. Methods: In this research, we evaluated the potential cardiotonic activity of six new synthesized selective PDE3 inhibitors (6-hydroxy-4-methylquinolin-2(1H)-one derivatives) using the spontaneously beating atria model. In each experiment, atrium of reserpine-treated rat was isolated and the contractile and chronotropic effects of a synthesized compound were assessed. The 3-isobutyl-1-methylxanthine, a non-selective PDE inhibitor, was used for comparison. Results: The results showed that, among new compounds, the best pharmacological profile was obtained with the compound 6-[4-(4-methylpiperazine-1-yl)-4-oxobutoxy]-4-methylquinolin-2(1H)-one, C6, which displayed selectivity for increasing the force of contraction (168 +/- 5% change over the control) rather than the frequency rate (138 +/- 5% change over the control) at 300 muM. However, C6 at concentrations of 10 and 100 muM produced left and upward shift in the positive inotropic concentration-response curve of isoprenaline. The -log EC50 of isoprenaline was 8.843 +/- 0.171 in the absence, 9.448 +/- 0.138 and 9.456 +/- 0.107 in the presence of 10, 100 muM of C6, respectively (P<0.001, n = 6). Also, amrinone, a selective PDE3 inhibitor, shifted the isoprenaline concentration-response curve to the left and upward. The concentration of 10 and 100 muM amrinone decreased -log EC50 of isoprenaline to 9.527 +/- 0.287 and 9.423 +/- 0.243, respectively (P<0.001, n = 6). Moreover, the positive chronotropic effect of isoprenaline was not affected by amrinone or C6. Conclusion: This study provides functional evidence for the positive inotropic effect of C6. Considering the augmentation of isoprenaline positive inotropic concentration-response with C6 and amrinone, we conclude that C6 produces its effect via potentiation of cAMP-dependent signaling system and possibly by inhibition of PDE3 activity.     

Leishmania Parasite: the Impact of New Serum-Free Medium as an Alternative for Fetal Bovine Serum

Background:Flagellated protozoan of the genus Leishmania is the causative agent of vector-borne parasitic diseases of leishmaniasis. Since the production of recombinant pharmaceutical proteins requires the cultivation of host cells in a serum-free medium, the elimination of FBS can improve the possibility of large-scale culture of Leishmania parasite. In the current study, we aimed at evaluating a new serum-free medium in Leishmania parasite culture for future live Leishmania vaccine purposes. Methods:Recombinant L. tarentolae secreting PpSP15-EGFP and wild type L. major were cultured in serum-free (CSFM) and serum-supplemented medium. The growth rate, protein expression, and infectivity of cultured parasites in both conditions was then evaluated and compared. Results:Diff-Quick staining and epi-fluorescence microscopy examination displayed the typical morphology of L. major and L. tarentolae-PpSP15-EGFP promastigote grown in CSFM medium. The amount of EGFP expression was similar in CSMF medium compared to M199 supplemented with 5% FBS in flow cytometry analysis of L. tarentolae-PpSP15-EGFP parasite. Also, a similar profile of PpSP15-EGFP proteins was recognized in Western blot analysis of L. tarentolae-PpSP15-EGFP cultured in CSMF and the serum-supplemented medium. Footpad swelling and parasite load measurements showed the ability of CSFM medium to support the L. major infectivity in BALB/C mice. Conclusion:This study demonstrated that CSFM can be a promising substitute for FBS supplemented medium in parasite culture for live vaccination purposes. Key Words: Growth rate, L. major, L. tarentolae, Serum-free medium, PpSP15-EGFP protein     

Identifying Soil and Plant Nutrition Factors Affecting Yield in Irrigated Mature Pistachio Orchards

The main objective of this study was to evaluate the potential use of a hybrid Genetic Algorithm-Artificial Neural Network (GA–ANN) method for predicting pistachio yield and for identifying the determinant factors affecting pistachio yield in Rafsanjan region, Iran. A total of 142 pistachio orchards were selected randomly and soil samples were taken at three depths. Besides, water samples and leaves from branches without fruit were taken in each sampling point. Management information and pistachio yields were achieved by completing a questionnaire. Primarily, 58 variables affecting pistachio yield were measured, and then 26 out of them were selected by minimizing mean square error (MSE) using a feature selection (FS) method. The results showed that the accuracy of the method was acceptable. Furthermore, the sensitivity analysis showed that the main determinant features affecting the pistachio yield were the irrigation water amount, leaf phosphorus, soil soluble magnesium, electrical conductivity (EC), and leaf nitrogen.     

Spatial variability of soil microbial indices in common alder COMMON ALDER(Alnus glutinosa) stands using a geostatistical approach in northern Iran

Microbial indices and their spatial patterns are strongly affected by environmental factors. Spatial variability of soil properties is one of the most important causes of variability in soil microbial indices. This research was conducted in the Caspian forest to assess spatial variabilities and frequency distributions of microbial properties.Ninety soil samples were taken using a grid sampling design 40 9 40 m. Microbial indices, organic carbon,nitrogen and pH were determined. Soil variable distributions showed that microbial indices had abnormal distributions. Logarithmic transformation produced normal distribution. Spatial continuity using geostatistical(variogram) was studied and maps obtained by point kriging.The variograms revealed the presence of spatial autocorrelation. The results indicate that spatial dependence of soil microbial indices was affected by non-intrinsic factors and forest management procedures. The maps show that soil microbial indices and soil properties have spatial variability. The spatial pattern of microbial indices was correlated to organic carbon and nitrogen.     

Response of wheat growth and productivity to exogenous polyamines under lead stress

Polyamines [i.e. spermine (Spm), spermidine (Spd) and putrescine (Put)] antagonistic effects on stress imposed by 2.0 mM lead (Pb²⁺) on growth, yield and its components and changes in the osmoprotectant and endogenous Pb²⁺ concentrations, the contents of some nutrients and tissue health in wheat plants were evaluated. Under the three applied polyamine (PAs) applications, the efficiency of wheat plants to tolerate Pb²⁺ stress in terms of growth and yield characteristics was noticed to varying degrees. The enhancements in osmoprotectant concentrations and plant health [in terms of relative water content (RWC) and membrane stability index (MSI)], and reductions in electrolyte leakage (EL) and plant Pb²⁺ concentration were correlated with the reasonable growth of Pb²⁺-stressed plants and their grain yield. Results point out that, better growth and yield characteristics, MSI, RWC, leaf photosynthetic pigment and osmoprotectant concentrations, and nutrient contents were obtained with seed soaking in 0.25 mM Spm, 0.50 mM Spd or 1.0 mM Put than those generated with seed soaking in water under 2.0 mM Pb²⁺ stress. In contrast, EL and the concentration of endogenous Pb²⁺ were significantly reduced. However, the Pb²⁺-free control positively exceeded the all stressed treatments. Among all tested PAs, 1.0 mM Put showed the best results and thus is recommended, as seed soaking, for wheat to grow well under Pb²⁺ stress.     

Nitrate leaching in a silage maize field under different irrigation and nitrogen fertilizer rates

Quantification of the interactive effects of nitrogen (N) and water on nitrate (NO₃) loss provides an important insight for more effective N and water management. The goal of this study was to evaluate the effect of different irrigation and nitrogen fertilizer levels on nitrate-nitrogen (NO₃-N) leaching in a silage maize field. The experiment included four irrigation levels (0.7, 0.85, 1.0, and 1.13 of soil moisture depletion, SMD) and three N fertilization levels (0, 142, and 189 kg N ha⁻¹), with three replications. Ceramic suction cups were used to extract soil solution at 30 and 60 cm soil depths for all 36 experimental plots. Soil NO₃-N content of 0-30 and 30-60-cm layers were evaluated at planting and harvest maturity. Total N uptake (NU) by the crop was also determined. Maximum NO₃-N leaching out of the 60-cm soil layer was 8.43 kg N ha⁻¹, for the 142 kg N ha⁻¹ and over irrigation (1.13 SMD) treatment. The minimum and maximum seasonal average NO₃ concentration at the 60 cm depth was 46 and 138 mg l⁻¹, respectively. Based on our findings, it is possible to control NO₃ leaching out of the root zone during the growing season with a proper combination of irrigation and fertilizer management.     

Direct somatic embryogenesis in Epipactis veratrifolia,a temperate terrestrial orchid

Most temperate terrestrial orchids are endangered species. Attempts to produce plantlets from plants of the genus Epipactis by asexual methods have totally failed. This study was conducted using somatic embryogenesis as a rapid vegetative propagation technique for conservation of E. veratrifolia. For these purposes, effects of different types of plant growth regulators (PGRs), different types of explants, light and dark conditions, and the effect of gibberellic acid (GA₃) and Paclobutrazol (PBZ) were investigated on somatic embryogenesis induction. Abscisic acid (ABA) pre-treatment effectiveness on inducting somatic embryogenesis and increasing the number of embryos per explant were investigated. Subsequently, 16 media were tested to find the best medium for plant regeneration and shoot and root proliferation. BA (3 mg L^?1) resulted in a better response than the other PGRs by supporting the development of 17 embryos per protocorm. PBZ, which resulted in 11 embryos per explant, was better than GA₃. FAST medium supplemented with organic substances was recognized as the best medium for plant regeneration and shoot and root proliferation. ABA pre-treatment had positive effect on somatic embryogenesis initiation. This study, for the first time, succeeded in finding a rapid and suitable protocol for propagation and genetic resource conservation of E. veratrifolia.     

Effects of partial replacement of fish meal with Spirulina platensis meal in practical diets on growth,survival, body composition,and reproductive performance of three-spot gourami (Trichopodus trichopterus) (Pallas, 1770)

This study investigated the effects of fish meal replacement by Spirulina platensis meal on growth, survival, body biochemical composition, and reproductive performance of Trichopodus trichopterus until the first spawning for 16 weeks. The fish fed dietary algal meal at levels of 2.5, 5, 10, and 20 % against those received fish meal only as control. The results showed that indicators such as belly diameter, feed intake, feed conversion ratio (FCR), and viscerosomatic index were significantly different between treatments with 5 and 10 % algal replacements (P < 0.05). The growth factors including final weight, weight gain, total length, fork length, and body height of fish did not show significant differences (P > 0.05) between the treatments. Feed intake increased up to 10 % replacement level and then decreased, though the best FCR values (0.77–0.79) were obtained with 2.5 and 5 % substitutions. The body lipid content showed a statistically marked decrease (P < 0.05) as a result of the algal replacements, but the amounts of protein and moisture were not significantly (P > 0.05) different between treatments. All fish fed S. platensis meal had greater gonadosomatic indices (19.4–21.85 %) than that of the control in spite of insignificant differences (P > 0.05) in their ovary weights (2.07–2.21 g). Absolute fecundity ranged between 7300 and 12,700 eggs/female with the highest amount in fish fed with replaced S. platensis levels of 2.5–10 %. The second-order polynomial regression analysis between absolute fecundity and substitution levels of algal meal was y = ?0.5319x ² + 106.87x + 7812.9 (R ² = 0.9642), and the optimal substitution level was determined to be 8.13 % for maximum absolute fecundity. Hatching percentages in the control and 20 % replacement were much lower (P < 0.05) than those in the other treatments (78–86 %). Given the relative enhancement of growth parameters and significant improvement in reproductive performance of broodstock three-spot gourami, 8.1–9.6 % S. platensis meal instead of fish meal is recommended in the diets.     

Immobilization of Urease onto Modified Egg Shell Membrane through Cross Linking

Background:Immobilization is an approach in industry to improve stability and reusability of urease. The efficiency of this technique depends on the type of membrane and the method of stabilization. Methods:The PEI-modified egg shell membrane was used to immobilize urease by absorption and glutaraldehyde cross-linking methods. The membranes were characterized by FTIR and AFM, and Nessler method was applied to measure the kinetic of the immobilized enzymes. Finally, the storage stability (6 °C for 21 days) and reusability (until enzyme activity reached to zero) of the immobilized enzymes were investigated. Results:Based on FTIR, three new peaks were observed in both the absorption- (at 1389.7, 1230.8, and 1074.2 cm^-1) and the cross-linking (at 1615-1690, 1392.7, 1450 cm^-1) immobilized enzymes. The surface roughness of the native membrane was altered after PEI treatment and enzyme immobilization. The optimal pH of cross-linking immobilized enzymes was shifted to a more neutral pH, while it was alkaline in adsorption-immobilized and free enzymes. The reaction time decreased in all immobilized enzymes (100 min for free enzyme vs. 60 and 30 min after immobilizing by adsorption and cross-linking methods, respectively). The optimal temperature for all enzymes was 70 °C and they had a higher K_m and a lower V_max than free enzyme. The stability and reusability of urease were improved by both methods. Conclusion:Our findings propose these approaches as promising ways to enhance the urease efficiency for its applications in industries and medicines. Key Words: Egg shell, Immobilization, Polyethylenimine, Urease