Biochemical and Biological Evaluation of an L-Asparaginase from Isolated Escherichia coli MF-107 as an Anti-Tumor Enzyme on MCF7 Cell Line

Background: One of the most widely used anticancer agents is microbial L-ASNase. Herein, we assessed the biochemical and biological properties of an isolated L-ASNase from a Gram-negative bacteria strain, Escherichia coli MF-107. Methods:Using garden asparagus, we obtained several bacterial isolates. These strains were further screened for L-ASNase activity. A promising bacterial isolate was selected for L-ASNase production and subsequent purification. The molecular weight of purified L-ASNase was determined. The MTT assay was applied to assess the cytotoxic effect of the purified enzyme. Also, for caspase activity determination and the apoptotic effect of purified enzyme on in cells, we conducted a real-time PCR method. Results:The molecular weight of the enzyme was approximately 37 kDa. In the pH range of 7.5 to 8, the enzyme had considerable stability. At 35 °C, the purified L-ASNase optimum activity was recorded. The cytotoxic effect of the enzyme on treated cells was dose-dependent with an IC₅₀ value of 5.7 IU/ml. The Bax gene expression considerably raised by 5.75-fold (p < 0.001) upon L-ASNase treatment. On the other hand, the anti-apoptotic Bcl-2 gene expression showed a 2.63-fold increase compared to the control (p < 0.05). It was detected that the mRNA levels of caspase-3 and p53 were considerably upregulated (5.93 and 1.85-fold, respectively). We did not find any alternation in the caspase-8 activity of the treated cells compared to untreated cells. Conclusion:In this research, the proliferation of the breast cancer cells remarkably inhibited via the cytotoxic effect of isolated L-ASNase from microbial sources.Key Words: Apoptosis, Breast cancer, Escherichia coli, MCF7 cell line     

Wide hybridization and introgression breeding in safflower: Effectiveness of different selection methods

Possibility of using interspecific hybridization to increase genetic diversity of drought tolerance in safflower (Carthamus tinctorius L.) and effectiveness of different selection methods is poorly understood. The main objectives of this study were to evaluate the effectiveness of (a) interspecific hybridization to expand genetic diversity in safflower for agronomic traits and drought tolerance in advance generations and (b) different selection methods (direct vs. indirect) for improving seed yield. Interspecific hybridizations were performed between species of C. tinctorius × C. palaestinus (TP), C. palaestinus × C. oxyacantha (PO) and C. tinctorius × C. oxyacantha (TO). Based on the field screening of genotypes in F3 generation, four groups of selected genotypes (derived from combination of direct and indirect selection under both drought and normal environments along with selection based on drought tolerance index (STI)) were evaluated in F4 and F5 generations. Indirect selection for seed yield was significantly more efficient than direct selection. Selection in stress environment resulted to more efficiency than selection in normal environment. No significant difference was observed between direct selection and selection based on STI. The result showed that TP was the best population to combine high seed yield and drought tolerance in safflower.     

Colonization with endo-mycorrhiza affects the resistance of safflower in response to salinity condition

Two Safflower (Carthamus tinctoriusL.) cultivars' seeds were used to study the influence of inoculation with mycorrhiza arbuscular fungi under salinity stress condition. Factorial experiment based on completely randomized design (three-way analysis of variance (ANOVA)) with 3 replications was used. Salinity treatment with 3 levels (0.5, 6, and 12 dS/m) and mycorrhizal arbuscular inoculation with two species (three levels consist of non-inoculation, and inoculation with Glomus intraradices and G. moseae) were applied on two cultivars of safflower (Goldasht and Padide) in this experiment. Some important biochemical, mineral, and growth traits were measured in this study. Salinity had a significant negative effect on all growth and morphological parameters including shoot and root dry weight, stem and root height. On the other hand, colonized plants showed better growth parameters under saline conditions compared with the control. The colonization of both mycorrhizal species decreased with salinity. Sugar and pigment content decreased with salinity, but their levels in colonized plants under saline conditions were higher than that in non-colonized plants. Mineral elements including phosphorus (P), nitrogen (N), and magnesium (Mg) were higher in colonized plants, while salinity decreased the absorption of these elements in both inoculated and non-inoculated plants. Higher activity of the enzymatic antioxidant means higher removal of these compounds and higher resistance to stress condition. Overall, it is clear that salinity had a negative effect on both cultivars of safflower, but these negative effects were lower in inoculated plants than in the non-inoculated ones; so, the use of mycorrhizal inoculation is a proper way to control the effect of salinity and maintain plant production.     

Comparative analysis of 31 reference evapotranspiration methods under humid conditions

Evaluation of simple reference evapotranspiration (ET_o) methods has received considerable attention in developing countries where the weather data needed to estimate ET_o by the Penman–Monteith FAO 56 (PMF-56) model are often incomplete and/or not available. In this study, eight pan evaporation-based, seven temperature-based, four radiation-based and ten mass transfer-based methods were evaluated against the PMF-56 model in the humid climate of Iran, and the best and worst methods were selected from each group. In addition, two radiation-based methods for estimating ET_o were derived using air temperature and solar radiation data based on the PMF-56 model as a reference. Among pan evaporation-based and temperature-based methods, the Snyder and Blaney–Criddle methods yielded the best ET_o estimates. The ET_o values obtained from the radiation-based equations developed here were better than those estimated by existing radiation-based methods. The Romanenko equation was the best model in estimating ET_o among the mass transfer-based methods. Cross-comparison of the 31 tested methods showed that the five best methods as compared with the PMF-56 model were: the two radiation-based equations developed here, the temperature-based Blaney–Criddle and Hargreves-M4 equations and the Snyder pan evaporation-based equation.     

Applicability of support vector machines and adaptive neurofuzzy inference system for modeling potato crop evapotranspiration

Estimation of crop evapotranspiration (ET_C) for certain crops such as potato is very important for irrigation planning, irrigation scheduling and irrigation systems management. The primary focus of this study was to investigate the accuracy of the adaptive neurofuzzy inference system (ANFIS) and support vector machines (SVM) for potato ET_C estimation when lysimeter measurements or the complete weather data for applying the FAO method are not available. The estimates of the ANFIS and SVM models were compared with the empirical equations of Blaney–Criddle, Makkink, Turc, Priestley–Taylor, Hargreaves and Ritchie. The performances of the different SVM and ANFIS models were evaluated by comparing the corresponding values of root mean square error (RMSE), mean absolute error (MAE) and correlation coefficient (r). The drawn conclusions confirmed that the SVM and ANFIS models could provide more accurate ET_C estimates than the empirical equations. Overall, the minimum RMSE (0.042 mm/day) and MAE (0.031 mm/day) values and the maximum r value (0.98) were obtained using the SVM model with mean air temperature, relative humidity, solar radiation, sunshine hours and wind speed as inputs.     

Evaluation of aqueous and ethanolic extracts of saffron, Crocus sativus L., and its constituents, safranal and crocin in allodynia and hyperalgesia induced by chronic constriction injury model of neuropathic pain in rats

The current study was designed to evaluate therapeutic potential of systemically administered ethanolic and aqueous extracts of saffron as well as its bioactive ingredients, safranal and crocin, in chronic constriction injury (CCI)-induced neuropathic pain in rats. The von Frey filaments, acetone drop, and radiant heat test were performed to assess the degree of mechanical allodynia, thermal allodynia and thermal hyperalgesia respectively, at different time intervals, i.e., one day before surgery and 3, 5, 7 and 10 days post surgery. The ambulatory behavior was evaluated using the open field test. A 7-day treatment with the ethanolic and aqueous extracts (50,100 and 200 mg/kg, i.p.) and safranal (0.025, 0.05 and 0.1 mg/kg, i.p.), attenuated the behavioral symptoms of neuropathic pain in a dose dependent manner. Crocin even at the high dose (50 mg/kg) failed to produce any protective role. However, gabapentine (100 mg/kg) as a reference drug significantly alleviated all behavioral manifestations of neuropathic pain compared to control group. In conclusion, the results of this study suggest that ethanolic and aqueous extracts of saffron as well as safranal could be useful in treatment of different kinds of neuropathic pains and as an adjuvant to conventional medicines.     

The true nature of Ganoderma in Iran: Taxonomy based on ITS and mtSSU rDNA

The genus Ganoderma Karst. has broad‐spectrum usage in biotechnology, medicine and the food industry. The complexity of the morphology within species has led to uncertain identification in the past, but recent advancements in molecular identification methods have provided scientists with the opportunity to better understand the taxonomy of the species. The present study attempts for the first time to elucidate the distinctiveness of the Ganoderma species growing in Iran concerning those elsewhere in the world based on mitochondrial small subunit ribosomal DNA (mtSSU rDNA) and internal transcribed spacer (ITS) sequence data. The results disclosed that the G. lucidum Karst. samples collected in Iran are more similar to the European Ganoderma species than to the Asian (Chinese) one used in this study.     

Isolation and identification of effective cellulolytic bacteria in composting process from different sources

Cellulolytic bacteria have a bio-activating role in the composting process. A study was carried out to isolate and identify cellulolytic bacteria from various sources. The isolates were cultured in the carboxymethyl cellulose (CMC) agar medium and incubated at 30°C for 3–7 days. Based on morphological characteristics of the isolates, maximum diameter of a clear zone around the colony and maximum cellulolytic activity, eight isolate were selected for further studies regarding composting experiments.

Molecular tests based on PCR amplification and sequencing of 16S rRNA gene of isolates showed the closest phylogenetic similarity with the species of Stenotrophomonas rhizophila DSM14405 (99.8%), Brevibacterium halotolerans DSM8802 (99.6%), Achromobacter marplatensis B2 (99.8%), Bacillus methylotrophicus CBMB205 (100%), Pseudomonas azotoformans IAM 1603 (99.7%), Bacillus sonorensis NBRC 101234 (99.8%), Bacillus subtilis KCTC 13429 (100%) and Ochrobactrum thiophenivorans DSM 7216 (99.3%). The study of the isolates impact on the composting of palm wastes in a randomized complete block design with 11 treatments in 3 replications showed that strain IB (B. methylotrophicus) caused a significant decrease in C:N ratio (58%). The increasing of microbial respiration compared with control after 30 days incubation at 37°C showed that the B. methylotrophicus strain IB with cellulolytic characteristics can be applied to hydrolysis of cellulosic biomass in the composting processes.     

Integrated pest management of Tuta absoluta: practical implementations across different world regions

Journal of Pest Science - The South American tomato pinworm, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), has invaded most Afro-Eurasian countries and is threatening worldwide tomato...