Study of structure-activity relationship in Aurein 1.2 analogs

Two new analogs of Aurein 1.2 antimicrobial peptide were synthesized and the antimicrobial activities were investigated. The results showed that the activity of G1R/F3W analog was higher than the native peptide and the F3W analog. Circular dichroism studies also showed that the secondary structure of the F3W was concentration-dependent, whereas, there was no such relationship seen in the case of G1R/F3W analog. It has been proposed that G1R/F3W activity was based on a single mechanism (snorkeling), while Aurein 1.2 and F3W utilized the snorkeling mechanism at low concentrations (0-0.01 mM) and the carpet mechanism at higher concentrations (0.01-0.1 mM). This study suggests that one pay attention to the concentration of biomolecules in peptide-based drug design.     

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Background: Pseudomonas aeruginosa is one of the opportunistic pathogens causing frequent hospital-acquired life-threatening infections in mechanically ventilated patients. The most significant virulence factor of P. aeruginosa is T3SS. PcrV is an important structural protein of the T3SS. Methods:In the current investigation, a recombinant scFv mAb against the PcrV protein was expressed in EnBase® (fed-batch) cultivation mode. The pETiteTM N-His SUMO Kan vector, including anti-PcrV scFv gene, was transformed into Escherichia coli (BL21) cells. The expression and solubility of anti-PcrV scFv protein were investigated at two different temperatures (25 °C and 30 °C) and at different induction times (4, 6, 8, 12, and 24 hours). Results:Increased efficiency was achieved by EnBase® compared to LB broth; owing to the slow release of glucose, the maximum level of solubility and total protein expression was observed in EnBase® cultivation system at 30 °C and 24 h post induction. Furthermore, IC₅₀ for anti-PcrV scFv protein was determined to be approximately 7 μg/mL. Conclusion:Anti-PcrV scFv produced in this study showed promising in vitro results, protecting RBC from lysis by P. aeruginosa (exoU+). Key Words: Fed Batch, recombinant protein, Pseudomonas aeruginosa, scFv     

Evaluation of genetic diversity among some Iranian wild asparagus populations using morphological characteristics and RAPD markers

An evaluation of genetic diversity in 39 wild asparagus populations was carried out using morphological and RAPD markers. A combination of morphological traits and random RAPD primers was used to examine the level of genetic variation and polymorphisms among the populations. A factor analysis using Ward's method on mean values of morphological characteristics indicated seven main factors resulting in four groups. Analysis of polymorphic bands using Jaccard's similarity coefficient indicated that genetic similarity ranged between 0.71 and 0.29. At a similarity level of 0.64, the populations were divided in three sub-clusters, containing 34, four and one populations, respectively. Significant regression associations were found between 21 morphological characteristics and 18 RAPD markers, revealing some informative markers associated with some traits. The highest R² was related to 18 RAPD markers associated with gender (53.5%) that among them BA-04₂₀₀₀ had a maximum R². The results showed that Iranian wild asparagus with its high levels of genetic variation could be considered as a valuable gene pool for future asparagus breeding programs. Furthermore, it could be inferred that morphological characteristics and RAPD markers are suitable tools to discriminate asparagus populations for the evaluation of genetic diversity.     

Impact of some alternative methods to chemical control in controlling aphids (Hemiptera: Sternorrhyncha) and their side effects on natural enemies on young Moroccan citrus groves

Aphid control in Moroccan citrus orchards is based mainly on carbamate and neonicotinoid sprays, especially methomyl and imidacloprid. The extensive use of these insecticides may have side effects on natural enemies and environment quality and raises human health concerns. This research aimed to assess the control of aphids with insecticidal soap, kaolin and augmentative biological control using the indigenous predator Adalia decempunctata L. (Coleoptera: Coccinellidae). The insecticides were applied and the predators were released in April 2009 and 2010. Under field experimentation, the methomyl and imidacloprid foliar pulverization were very effective against aphids. In contrast, the insecticidal soap and kaolin application were less efficient while A. decempunctata adults were effective only in the first week after release. The side effects on beneficial insects were also assessed and discussed. The possibility of employing A. decempunctata in an integrated pest management package in citrus groves is discussed in relation to effectiveness and side effects on beneficial arthropods.     

Effects of the dietary protein levels and the protein to energy ratio in sub‐yearling Persian sturgeon,Acipenser persicus (Borodin)

A 3 × 4 factorial design was used to evaluate the dietary protein requirement and to determine the optimum dietary protein to energy (P/DE) ratio in sub‐yearling Persian sturgeon, Acipenser persicus, reared in the indoor system. Twelve experimental diets (₄₀P₁₆, ₄₀P₁₇, ₄₀P₁₈, ₄₀P₁₉, ₄₅P₁₉, ₄₅P₁₇, ₄₅P₁₈, ₄₅P₁₉, ₅₀P₁₆, ₅₀P₁₇, ₅₀P₁₈ and ₅₀P₁₉) were formulated and prepared to contain three protein levels (40%, 45% and 50%) and four digestible energy levels (16, 17, 18 and 19 kJ g^?1 diet) at each protein level. Fish averaging 103.3 ± 3.5 (mean ± SD) were fed one of the experimental diets for 14 weeks. At the end of the experimental period, there were significant energy effects (P < 0.05) on weight gain (WG) and specific growth rate (SGR). Weight gain and SGR tended to decrease, although non‐significantly, with increase in dietary protein levels. Furthermore, there were significant protein and energy interaction effects on WG, SGR, hepatosomatic index and protein efficiency ratio. However, there were no significant dietary protein, energy or their interaction effects on feed efficiency for fish fed all diets. Weight gain and SGR of fish fed ₄₀P₁₉ were significantly higher than those of fish fed ₄₀P₁₆, ₄₅P₁₆, ₄₅P₁₇, ₅₀P₁₆ and ₅₀P₁₇ diets (P < 0.05). There were no significant differences in WG and SGR among fish fed ₄₀P₁₇, ₄₀P₁₈, ₄₀P₁₉, ₄₅P₁₈, ₄₅P₁₉, ₅₀P₁₈ and ₅₀P₁₉ diets. These results may indicate that the optimum dietary protein requirement and the P/DE ratio could be 40% protein and 22.0 mg protein kJ^?1 (₄₀P₁₈), respectively, in Persian sturgeon, based on growth performance and feed utilization.     

Chrysin Reduced Acrylamide-Induced Neurotoxicity in Both in vitro and in vivo Assessments

Background: Acrylamide (ACR) is a well-known industrial toxic chemical that produces neurotoxicity, which is characterized by progressive central and peripheral neuronal degeneration. Chrysin is a natural, biologically active flavonoid compound, which is commonly found in many plants. The antioxidant and neuroprotective properties of chrysin have been demonstrated. Methods: In this study, the possible effect of chrysin on ACR-induced toxicity was evaluated in both in vitro and in vivo experiments. PC12 cells were used as a suitable in vitro model. Cells were exposed to chrysin (0.5-5 µM) for 12 and 24 h, and then ACR in IC₅₀ concentration was added to the cells. Finally, cell viability was determined using (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium assay. For in vivo assay, Wistar rats were treated with ACR (50 mg/kg i.p. for 11 days) alone or in combination with chrysin (12.5, 25, and 50 mg/kg). At the end of treatment, behavioral index was evaluated. Results: ACR decreased cell viability and pre-treatment with chrysin (0.5-5 µM) significantly decreased ACR-induced cytotoxicity in the time- and dose-dependent manner. In Wistar rats, exposure to ACR significantly induced severe gait abnormalities, but treatment with chrysin (50 mg/kg) reduced ACR-induced neurotoxicity in animals. Conclusion: In the current study, chrysin exhibited neuroprotective effect on PC12 cells as an in vitro model and also on Wistar rats. Iran. Key Words: Acrylamide, Chrysin, Neurotoxicity, Antioxidant     

Volatile components of Camellia sinensis inhibit growth and biofilm formation of oral streptococci in vitro

This study aimed to evaluate the efficacy of semi fermented and non fermented Camellia sinensis extracts (Black and Green tea) and comparison between them against Streptococcus mutans ATCC 25175, S. mitis ATCC 9811 and S. sanguis ATCC 10556 that are responsible for dental caries and bacteremias following dental manipulations. Minimum inhibitory concentrations of both tea extracts were assessed by Well diffusion and Broth dilution methods and examination of cell adherence (Biofilm inhibitory concentrations) was observed on glass slides under phase contrast microscope and colony counts from glass beads. Concentration of 1 mg mL(-1) of semi fermented tea extract was completely biofilm inhibitor but biofilm formation by these bacteria was seen 7 days after treatment with 1 mg mL(-1) of non fermented Camellia sinensis on glass beads and BIC for oral streptococci treated with this extract was 1.5, 2.5 mg mL(-1) of semi fermented and 3 mg mL(-1) of non fermented extracts had bactericidal effect on these bacteria. Semi fermented and non fermented Camellia sinensis extracts were able to prevent growth of oral streptococci. Therefore dental caries significantly reduce and the efficiency of semi fermented tea was higher due to rich content of volatile components rather than non fermented extracts.     

Characterisation of a ω-gliadin gene in Triticum tauschii

A ω-gliadin gene at the Gli-D^t1 locus of Triticum tauschii accession AUS18913 was isolated using PCR primers, designed from published sequences of ω-gliadin genes of bread wheat cv Cheyenne, and deduced sequences of the N-terminal amino acids of ω-gliadin proteins. Further, the derived protein was isolated from A-PAGE and was sequenced. The protein sequence contained a signal peptide of 19 amino acids followed by a short N-terminal sequence of 11 amino acids, a central repetitive domain that covers approximately 90% of the sequence and a short C-terminal domain of 12 amino acids. The sequence comparison with other ω-gliadins showed a high level of similarities between them. Further analysis of the ω-gliadins using A-PAGE revealed that there are three ω-gliadin proteins in AUS18913 accession. Comparison of N-terminal sequences of these proteins revealed that two of these proteins have very high homologies with ω-gliadins of Cheyenne while the third one was significantly different.     

YIELD AND FRUIT QUALITY OF ALMOND,PEACH AND PLUM UNDER REGULATED DEFICIT IRRIGATION

• Regulated deficit irrigation was assessed in almond, peach and plum over 3 years.• Fruit-growth slowdown stages are appropriate periods to apply deficit irrigation.• Peach yields were unaffected under a regulated deficit irrigation of 75% ET_C.• Regulated deficit irrigation of 50% ET_C maintained yields of almond and plum.• Fruit quality improved under regulated deficit irrigation.The effects of regulated deficit irrigation (RDI) on the performance of almond cv. Tuono, peach cv. JH-Hall and plum cv. Stanley were assessed on the Saiss Plain (NW, Morocco) over three consecutive growing seasons (2011–2013). Irrigation treatments consisted of a control, irrigation applied to fully satisfy crop water requirements (100% ET_C), and two RDI treatments, irrigation applied to 75% ET_C (RDI-75) and 50% ET_C (RDI-50). These three treatments were applied during fruit-growth slowdown periods corresponding to Stages II and III in almond and Stage II in peach and plum. Yield and fruit quality traits were determined. The effect of RDI differed between species. Yield and fruit size were reduced significantly only in peach under RDI-50. Fruit quality improved in this species in the first year of the experiment, with an increase of sugar/acid ratio and polyphenol content. Plum quality also improved but the effects were significant only in the second and third years. Similar results were recorded in almond kernel, but their epidermal grooves were deeper under RDI-50, and this may have affected their commercial value. It is concluded that water can be saved during the fruit-growth slowdown period by up to 25% in peach and 50% in almond and plum with improvements in fruit quality without affecting total yield.