Mitochondrial NAD(P)-dependent malic enzyme from herring testicular tissue: Purification,kinetic behaviour and regulatory properties

Mitochondrial NAD(P)-dependent malic enzyme [EC 1.1.1.39, L-malate: NAD⁺ oxidoreductase (decarboxylating)] was purified from herring testicular tissue to a specific activity of 26.4 μmol NADH/min/mg protein. Herring testicular tissue is one of the most abundant sources of this enzyme. The purification procedure involved differential centrifugation of mitochondria and then chromatography on DEAE-Sephacel, Red Agarose and Sephacryl S-300. This enzyme catalyzes the oxidative decarboxylation of malate in the presence of Mn²⁺ and either NAD or NADP. Under V_max conditions the ratios for the rate of NAD/NADP reduction was 1.8. A study of the reductive carboxulation reaction indicated that this enzyme reaction is reversible; at pH 7.0 the reverse reaction exhibited 22% of the activity of forward reaction. Some kinetic characteristics of the enzyme were determined. ATP was found to be a competitive inhibitor with respect to malate. Fumarate reversed ATP inhibition. Regulation of NAD(P)-dependent malic enzyme from herring testicular tissue mitochondria could respond to changing levels of mitochondrial ATP and fumarate in vivo.     

Effects of green tea polyphenol (-)-epigallocatechin-3-gallate on newly developed high-fat/Western-style diet-induced obesity and metabolic syndrome in mice

The aim of this study was to investigate the effects of (-)-epigallocatechin-3-gallate (EGCG) on newly developed high-fat/Western-style diet-induced obesity and symptoms of metabolic syndrome. Male C57BL/6J mice were fed a high fat/Western-style (HFW; 60% energy as fat and lower levels of calcium, vitamin D(3), folic acid, choline bitartrate, and fiber) or HFW with EGCG (HFWE; HFW with 0.32% EGCG) diet for 17 wks. As a comparison, two other groups of mice fed a low-fat diet (LF; 10% energy as fat) and high-fat diet (HF; 60% energy as fat) were also included. The HFW group developed more body weight gain and severe symptoms of metabolic syndrome than the HF group. The EGCG treatment significantly reduced body weight gain associated with increased fecal lipids and decreased blood glucose and alanine aminotransferase (ALT) levels compared to those of the HFW group. Fatty liver incidence, liver damage, and liver triglyceride levels were also decreased by the EGCG treatment. Moreover, the EGCG treatment attenuated insulin resistance and levels of plasma cholesterol, monocyte chemoattractant protein-1 (MCP-1), C-reactive protein (CRP), interlukin-6 (IL-6), and granulocyte colony-stimulating factor (G-CSF). Our results demonstrate that the HFW diet produces more severe symptoms of metabolic syndrome than the HF diet and that the EGCG treatment can alleviate these symptoms and body fat accumulation. The beneficial effects of EGCG are associated with decreased lipid absorption and reduced levels of inflammatory cytokines.     

Extraction of newly immobilized n from an illinois mollisol using aqueous phenol

Studies were conducted on an ¹⁵N-enriched Illinois Mollisol (Flanagan silt loam) to determine the extractability of newly immobilized N with aqueous phenol (45%, v/v). In comparison to mild extradants proposed as indexes of plant available N (i.e. 10 mw NaHCO₃ and hot 10 m CaCl₂), phenol showed the highest selectivity for the immobilized N, both with regard to the amount of biomass N extracted and the magnitude of the extractability ratio (ER). Aqueous phenol was also more selective in removing the immobilized N than reagents used for extraction of soil organic matter (i.e. 150 m Na₄P₂O₇ and 100 m KOH). Results with soils incubated for up to 120 days showed that the amount of ¹⁵N extracted with aqueous phenol fluctuated with time and was related to changes in the soil microbial population, e.g. ER was highest when the bacterial population was maximal. Aqueous phenol shows promise as a selective extradant of potentially-available (labile) organic N in soils.     

Production of Lectins from Marine Algae: Current Status,Challenges, and Opportunities for Non-Destructive Extraction

Marine algae are an excellent source of novel lectins. The isolation of lectins from marine algae expands the diversity in structure and carbohydrate specificities of lectins isolated from other sources. Marine algal lectins have been reported to have antiviral, antitumor, and antibacterial activity. Lectins are typically isolated from marine algae by grinding the algal tissue with liquid nitrogen and extracting with buffer and alcohol. While this method produces higher yields, it may not be sustainable for large-scale production, because a large amount of biomass is required to produce a minute amount of compound, and a significant amount of waste is generated during the extraction process. Therefore, non-destructive extraction using algal culture water could be used to ensure a continuous supply of lectins without exclusively disrupting the marine algae. This review discusses the traditional and recent advancements in algal lectin extraction methods over the last decade, as well as the steps required for large-scale production. The challenges and prospects of various extraction methods (destructive and non-destructive) are also discussed.     

Developmental temperature stress and parental identity shape offspring burst swimming performance in sockeye salmon (Oncorhynchus nerka)

Abstract – The persistent effects of embryonic temperature stress and individual parentage on fry swimming performance were examined in a cross‐fertilisation experiment using sockeye salmon (Oncorhynchus nerka). A fixed‐velocity test of burst swimming was used to assess the endurance capacity and behavioural performance of individual fry from 10 offspring families incubated at 12, 14 or 16 °C to hatch and then reared through yolk absorption and exogenous feeding stages in a common posthatch environment (average 6.9 °C). Fry burst swim time (BST) was influenced by an interaction between incubation temperature and family identity. Average BST was longer for fry from the 12 °C prehatch treatment compared to 14 and 16 °C, although differences were largely attributable to temperature effects on average fry size. Behavioural observations revealed that fish incubated at 16 °C performed more poorly, having a larger proportion of individuals that required stimulation to swim, fatigued more frequently or were classified as ‘nonswimmers’. Within all three incubation temperature treatments, mean BST varied significantly among offspring families, independent of fry mass and length. An interesting relationship was observed within the 16 °C treatment, whereby families with higher survivorship were characterised with lower mean BSTs. Collectively, these findings demonstrate that exposure to high temperatures in early sockeye salmon development can result in persistent, parentally mediated effects on fry performance. As such, these results provide important insight into how elevated temperature events during egg incubation may affect early life history selection processes and survival in stages beyond when the stressor is experienced.     

Influence of undegraded intake protein on reproductive performance of primiparous beef heifers maintained on stockpiled fescue pasture

The objectives of this study were to evaluate the effects of pre- and postpartum undegraded intake protein (UIP) supplementation on body condition score (BCS), BW, calf weight, milk production, serum IGF-I concentrations, and postpartum interval in primiparous beef heifers (n = 44). Heifers were maintained on endophyte-free stockpiled tall fescue (11.7% CP, 38% ADF) and individually fed supplement daily beginning 60 d prepartum. Pre- and postpartum supplements provided 19.3% CP, 83.4% TDN (UIP); 14.1% CP, 84.1% TDN (Control); 21.5% CP, 81.5% TDN (UIP); and 14.6% CP, 81.4% TDN (Control); respectively. Blood meal (146 g/d) was the source of UIP. Six heifers were removed from the study due to calf loss unrelated to treatment; therefore, postpartum measurements are based on 19 animals per treatment. Statistical analyses using ANOVA and a split-plot design revealed no effects of treatment (P > 0.2) on BCS, BW, calf weight, milk production, or postpartum interval. There tended to be a treatment x time interaction on BCS (P < 0.09) with UIP heifers having higher BCS than Control at wk 5, 7, and 9 postpartum. There was a treatment x time interaction on serum IGF-I (P < 0.06) during the first 35 d postpartum. In UIP heifers, serum IGF-I was greater at calving compared with Control heifers (117.5 vs 92.4 ng/mL, respectively); however, these differences were not related to changes in BCS or BW. Although serum IGF-I concentrations were increased at calving in heifers receiving UIP, there were no treatment effects on postpartum interval (P > 0.7). During the first 30 d postpartum, IGF-I differed (P < 0.01) among heifers with postpartum intervals defined as short, < 50 d (128.9 ng/mL); medium, 51 to 65 d (115.2 ng/mL); and long, 66 to 130 d (52.9 ng/mL). When analyzed as a regression, a 1 ng/mL increase in IGF-I (UIP and Control heifers) at calving (P < 0.05) and throughout the postpartum period (P < 0.01) corresponded to a decrease in postpartum interval of 0.13 d. Based on the results of this study, the inclusion of UIP in diets for primiparous heifers and its effects on postpartum interval warrant further evaluation.