Quercetin attenuates H2O2‐induced toxicity of rooster semen during liquid storage at 4°C

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H₂O₂ (40 μM) and combination groups which incubated with constant dose of H₂O₂ (40 μM) plus various levels of quercetin (20, 40 and 80 μM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 μmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 μmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 μmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co‐administration of quercetin (especially at 40 and 80 μM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide‐mediated damage during cold liquid storage of rooster semen.     

Sperm motility and seminal plasma characteristics in Barbus sharpeyi (Günther, 1874)

Spermatozoa concentration, ionic composition, osmolality, glucose and total protein contents of seminal plasma and sperm motility were determined in Barbus sharpeyi (Cyprinidae, Teleosotei). Spermatozoa concentration ranged from 9.77 to 20.20 × 10⁹ spermatozoa mL^?1. Osmolality (mOsmol kg^?1) and ionic contents (mM L^?1) of the seminal plasma were 274.5±9.0, 70.0±3.4 Na⁺, 28.8±0.9 K⁺, 101.7±3.1 Cl^?, 0.9±0.1 Mg²⁺ and 2.1±0.1 Ca²⁺ respectively. Total protein and glucose were 5.3±0.2 g L^?1 and 76.7±4.3 mM L^?1 respectively. Sperm motility was initiated in a hypo‐osmotic condition, composed of either an ionic (KCl or NaCl) or a non‐ionic (sucrose) activation medium. Duration of sperm motility was very short: <2 min after activation in distilled water. Percentage of motile spermatozoa was significantly higher in an activation medium containing NaCl compared with that of distilled water. An activating medium containing NaCl or KCl higher than 150 mM or sucrose higher than 275 mM totally inhibited the activation of sperm motility. Immediately after sperm activation, wave(s) propagated along the flagellum, but waves were restricted to the proximal part of the flagellum (close to the head) at 1 min post activation. Studied characteristics in the present study were compared with those of other cyprinids for understanding inter‐species differences.     

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     

Mucoadhesive Chitosan Electrospun Nanofibers Containing Tetracycline and Triamcinolone as a Drug Delivery System

The mucoadhesive Chitosan (CS) nanofibers as a drug delivery system were developed. Chitosan was modified via the immobilization of thiol groups from L-cysteine as a mucoadhesive reagent. The mucoadhesive properties of the chitosan nanofibers were evaluated by tensiometer set and via tensile studies. Drug and mucoadhesive agent loading lead to decrease diameters and increased porous of nanofibers. The release of Tetracycline (Tet) and Triamcinolone (Tri) were increased with increasing immersion time and it became constant at long immersion times. Mucoadhesion studies were done at pH 2–7 and in pH 6 maximum mucoadhesive properties observed. Release studies demonstrated a sustained release of both drug continued up to 48 hours. Microbial studies were performed on the nanofibers. The drug delivery system represented a novel tool for improve the therapeutic efficacy of various drugs that are poorly absorbed from the gastrointestinal tract. Also it is an efficient system for treatment of oral ulceration.     

Response of European seabass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) to canola oil diets: effect on growth performance,fish health and liver and intestine histomorphology

This study was undertaken to assess the effects of feeding European seabass (Dicentrarchus labrax) canola oil-added diets on growth, health status and liver and intestine histomorphology. Seabass (56.18 ± 0.16 g initial body weight) were fed one of three fish meal-based diets with ~48 % crude protein and ~16.0 % lipid, combining fish oil (FO) and canola oil (CO) for 12 weeks. The diets contained: zero (control, CTRL), 45 (CO50) or 63 (CO70) g CO kg^?1 assigned in triplicates to three dietary groups. The results indicated that neither dietary oil type (FO or CO) nor CO level adversely affected (P > 0.05) the growth, feed utilization or major blood constituents’ composition as an indicator of the overall health status of fish. Despite the CO diets influence on head kidney macrophage activity being unappreciable (P > 0.05), there was a reducing trend with an increase in CO level incorporation. The CO70 diet induced a minor fat infiltration in hepatocytes and leucocytes infiltration, hyperplasia of the basal nuclei and supranuclear vacuolization of the enterocytes of the distal intestine. The present observations suggest that it is possible to incorporate up to 63 g CO Kg^?1 in the feed for European seabass juveniles without major negative effects on growth, health status or liver and intestinal histomorphology.     

The Effects of Estrogen Receptors' Antagonist on Brain Edema,Intracranial Pressure and Neurological Outcomes after Traumatic Brain Injury in Rat

Background:

In previous studies, the neuroprotective effect of 17β-estradiol in diffuse traumatic brain injury has been shown. This study used ICI 182,780, a non-selective estrogen receptor antagonist, to test the hypothesis that the neuroprotective effect of 17β-estradiol in traumatic brain injury is mediated by the estrogen receptors.

Methods:

The ovariectomized rats were divided into eight groups. Brain injury was induced by Marmarou’s method. Estrogen was injected 30 minutes after traumatic brain injury, and ICI 182,780 was injected before traumatic brain injury and also before estrogen treatment. In one group only ICI 182,780 was injected. The brain water content and Evans blue dye content were measured 24 and 5 hours after traumatic brain injury, respectively. The neurologic outcomes and intracranial pressure were assessed before, 4, and 24 hours after traumatic brain injury.

Results:

Brain water content and Evans blue content were less in estrogen-treated group comparison to vehicle group. ICI 182,780 eliminated the effects of estrogen on brain edema and brain blood barrier permeability. Intracranial pressure was increased significantly after trauma, and estrogen decreased intracranial pressure at 4 and 24 hours after traumatic brain injury in comparison to vehicle. This inhibitory effect was also eliminated by treatment with ICI182,780. ICI 182,780 also inhibited the estrogen induced increase in neurologic outcomes following traumatic brain injury. However, the use of ICI 182,780 alone had no neuroprotective effect after traumatic brain injury.

Conclusion:

The results suggest that classical estrogen receptors have probably a role in the neuroprotective function of estrogen following traumatic brain injury.Key Words: Estrogens, Intracranial pressure (ICP), Brain edema     

Oxidative Stability of Spray-Dried Microencapsulated Fish Oils with Different Wall Materials

Multilayer microencapsulation of fish oil was evaluated by using fish gelatin (treated with or without microbial transglutaminase [MTGase]), chitosan, and maltodextrin for its ability to control lipid oxidation. This study showed that a mixture containing gelatin and maltodextrin obtained the highest percentage of core release. The oxidative stability of the formulated mixtures and the bulk oils was investigated during a period of 60 days. The peroxide value (PV) was assessed as a parameter for primary oxidation products, and p-anisidine (p-AV) and thiobarbituric acid reactive sub-stances (TBARS) was used to analyze secondary oxidation compounds. Observation of oxidation products showed that combinations of gelatin and maltodextrin made by adding MTGase and a mixture of gelatin and chitosan were able to increase the oxidative stability, and increases in PV and p-AV and TBARS were found for all oils.     

Stimulation in the movement and uptake of phosphorus in response to magnetic P solution and arbuscular mycorrhizal fungi in Ocimum basilicum

A greenhouse experiment was conducted to assess the effects of magnetic field, arbuscular mycorrhizal fungi (AMF), and phosphorus (P) concentration in the nutrient solution (0, 5, 10, 20, or 40 mg L^?1) on the mobility and accumulation of P in soil and plant tissues of basil (Ocimum basilicum L.). The experiment was designed as a factorial combination and treatments were arranged in a completely randomized design with four replicates. Magnetic field increased water-soluble P in the soil and P concentration in plant shoot by 30.0% and 13.0%, respectively, in comparison to the control. The application of magnetic field and inoculation of AMF at 10 mg P L^?1 increased the P translocation efficiency by 23.3% and 17.8%, respectively. Overall, our results demonstrated that the use of magnetic field and AMF could be an effective tool for enhancing of uptake and movement efficiency of P even at low concentrations.