Molecular characterization and identification of facilitative glucose transporter 2 (GLUT2) and its expression and of the related glycometabolism enzymes in response to different starch levels in blunt snout bream (<Emphasis Type="Italic">Megalobrama amblycephala</Emphasis>)

Facilitative glucose transporters (GLUT) are transmembrane transporters involved in glucose transport across the plasma membrane. In this study, blunt snout bream GLUT2 gene was cloned, and its expression in various tissues and in liver in response to diets with different carbohydrate levels (17.1; 21.8; 26.4; 32.0; 36.3; and 41.9% of dry matter). Blunt snout bream GLUT2 was also characterized. A full-length cDNA fragment of 2577 bp was cloned, which contains a 5′-untranslated region (UTR) of 73 bp, a 3′-UTR of 992 bp, and an open reading frame of 1512 bp that encodes a polypeptide of 503 amino acids with predicted molecular mass of 55.046 kDa and theoretical isoelectric point was 7.52. The predicted GLUT2 protein has 12 transmembrane domains between amino acid residues at 7–29; 71–93; 106–123; 133–155; 168–190; 195–217; 282–301; 316–338; 345–367; 377–399; 412–434; and 438–460. Besides, the conservative structure domains located at 12–477 amino acids belong to the sugar porter family which is the major facilitator superfamily (MFS) of transporters. Blunt snout bream GLUT2 had the high degree of sequence identity to four GLUT2s from zebrafish, chicken, human, and mouse, with 91, 63, 57, and 54% identity, respectively. Quantitative real-time (qRT) PCR assays revealed that GLUT2 expression was high in the liver, intestine, and kidney; highest in the liver and was regulated by carbohydrate intake. Compared with the control group (17.1%), fed by 3 h with higher starch levels (32.0; 36.3; and 41.9%), increased plasma glucose levels and glycemic level went back to basal by 24 h after treatment. Furthermore, higher dietary starch levels significantly increase GLUT2, glucokinase (GK), and pyruvate kinase (PK) expression and concurrently decrease phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6P) mRNA levels (P?<?0.05), and these changes were also back to basal levels after 24 h of any dietary treatment. These results indicate that the blunt snout bream is able to regulate their ability to metabolize glucose by improving GLUT2, GK, and PK expression levels and decreasing PEPCK and G6P expression levels.     

Influence of different microalgal diets on gonadal development of the carpet shell clam <Emphasis Type="Italic">Ruditapes decussatus</Emphasis> broodstock

Studies considering broodstock conditioning of Ruditapes decussatus mostly used flagellates and diatoms for feeding, mainly Isochrysis galbana and Chaetoceros calcitrans, respectively. The present study evaluated the effect of different microalgal diets on gonadic maturation of R. decussatus including three microalgae species (C. calcitrans, Tetraselmis suecica, and Nannochloropsis oculata) which were used to prepare six diets: three monospecific and three bispecific mixed diets at a proportion of 1:1. Clams were daily fed at a ratio of 1% dry weight of algae/live weight of clam and unfed treatment was used as control. Results showed that clams fed C. calcitrans alone or mixed with T. suecica attained earlier full maturity with the highest response to spawning induction (after 34 conditioning days) and largest proportion in the partial spawning phase for both, followed by T. suecica alone after 48 days. In contrast, N. oculata alone or mixed diets demonstrated poor performance for gonadal maturation. Unfed treatment did not show any sign of maturation. The results indicated that T. suecica has high conditioning potential like C. calcitrans and their mixture gave the highest benefit. Therefore, T. suecica can be recommended for R. decussatus broodstock conditioning.     

Preparation of Polyurethane Silicon Oxide Nanomaterials as a Binder in Leather Finishing

Leather finishing processes using toxic organic solvent based produce volatile organic compounds (VOC), chronic exposure to this chemicals effect on workers' health causing many diseases especially lung cancer. So, polyurethane waterbased was synthesized for application in leather finishing instead of organic solvent based because it’s economic and safety for industry and workers. Preparation of water-based polyurethane (PU) depends on the reaction of polyethylene glycol (PEG, 300) with isophorone diisocyanate (IPDI) and the reaction of IPDI-1,4-butanediol (BDO) together with dimethylolpropionic acid (DMPA), was synthesized by poly-addition polymerization reaction. PU was then modified with different amounts of silicon dioxide nanoparticles (1-5 % SiO₂), used as a binder in leather finishing. Leather coated was characterized physically, chemically and thermally by FTIR, GPC, DLS, TEM, SEM and TGA. The results revel that, water vapor permeability (WVP) of leather coated with PU modified with SiO₂ showed improvement due to the existence of SiO₂ particles which increases the interspaces of the polyurethane coating. SEM showed that when the amount of SiO₂ nanoparticles increases, there is uniform nanoparticles accumulated can be observed. EDX prove the presence of Si and O₂ elements and the formation of SiO₂ nanoparticles. Mechanical properties discussed that tensile strength; tear strength and elongation at break % increase with increase SiO₂ concentration until 3 % SiO₂ nanoparticles. TGA showed an improvement of thermal stability of coated leather modified with SiO₂. Therefore, this study succeeded in preparation of safe, ecofriendly of water-based polyurethane binders which modified with SiO₂ for using in leather finishing.     

Physicochemical and Biological Characterization of Fucoidan from Fucus vesiculosus Purified by Dye Affinity Chromatography

A comparative study concerning the physicochemical, monomeric composition and biological characters among different fucoidan fractions is presented. Common purification techniques for fucoidan usually involve many steps. During these steps, the important structural features might be affected and consequently alter its biological activities. Three purified fractions were derived from Fucus vesiculosus water extract which, afterwards, were purified by a recently-developed dye affinity chromatography protocol. This protocol is based on dye-sulfated polysaccharide interactions. The first two fractions were obtained from crude precipitated fucoidan at different pH values of the adsorption phase: pH 1 and 6. This procedure resulted in fucoidan_1 and 6 fractions. The other, third, fraction: fucoidan_M, however, was obtained from a buffered crude extract at pH 1, eliminating the ethanol precipitation step. All of the three fractions were then further evaluated. Results revealed that fucoidan_M showed the highest sulfur content (S%), 12.11%, with the lowest average molecular weight, 48 kDa. Fucose, galactose, and uronic acid/glucose dimers were detected in all fractions, although, xylose was only detected in fucoidan_1 and 6. In a concentration of 10 µg·mL⁻¹, Fucoidan_6 showed the highest heparin-like anticoagulant activity and could prolong the APTT and TT significantly to 66.03 ± 2.93 and 75.36 ± 1.37 s, respectively. In addition, fucoidan_M demonstrated the highest potency against HSV-1 with an IC₅₀ of 2.41 µg·mL⁻¹. The technique proved to be a candidate for fucoidan purifaction from its crude extract removing the precipitation step from common purification protocols and produced different fucoidan qualities resulted from the different incubation conditions with the immobilized thiazine toluidine blue O dye.     

Purification of glucose-6-phosphate dehydrogenase and glutathione reductase enzymes from the gill tissue of Lake Van fish and analyzing the effects of some chalcone derivatives on enzyme activities

Glucose-6-phosphate dehydrogenase (G6PD) and glutathione reductase (GR) are metabolically quite important enzymes. Within this study, these two enzymes were purified for the first time from the gills of Lake Van fish. In the purifying process, ammonium sulfate precipitation and 2′,5′-ADP Sepharose 4B affinity column chromatography techniques for glucose-6-phosphate dehydrogenase, temperature degradation and 2′,5′-ADP Sepharose 4B affinity column chromatography for glutathione reductase enzyme were used. The control of the enzyme purity and determination of molecular weight were done with sodium dodecyl sulfate polyacrylamide gel electrophoresis. K _M and V _max values were determined with Lineweaver–Burk plot. Besides, the effects of some chalcone derivatives on the purified enzymes were analyzed. For the ones showing inhibition effect, % activity–[I] figures were drawn and IC₅₀ values were determined. K _i value was calculated by using Cheng–Prusoff equation.     

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.     

Comparison of New Ultrasonic Digestion Approaches for Plant Matrices in the Analysis of Trace Metals by Inductively Coupled Plasma–Optical Emission Spectroscopy Analysis: Contrast with USEPA Method 3050B

An ultrasonic method using two approaches, A and B, along with a reference Environmental Protection Agency (EPA) Method 3050B [i.e., a mixture of 30 mL of nitric acid–hydrochloric acid–hydrogen peroxide–water (HNO₃-HCl-H₂O₂-H₂O)] were contrasted for leaching of a plant matrix. The trace metals were arsenic, cadmium, cobalt, chromium, copper, mercury, manganese, nickel, lead, selenium, and zinc (As, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Se and Zn) and quantified by ICP-OES followed by an investigation into residue formation and the impact of digestion time. Approach B was the most accurate and precise with percent recoveries ranging between 99 and 120%, whereas ultrasonic approach A and the USEPA method 3050B gave similar results with poor accuracies and precisions. In the optimization of the digestion time using approach B, the total metal recovery was fairly the same over a period of 120 min except for Cr and Cu, which showed slight variations.