Detection and localization of oxidized proteins in muscle cells by fluorescence microscopy

In meat, no detailed studies on the intracellular distribution of oxidized proteins during oxidative stress have been performed, to our knowledge. Therefore, we used fluorescence microscopy to detect and locate protein carbonyls, oxidation products of basic amino acids, generated in bovine M. Rectus abdominis during either exposition to a chemical free radical generating system, or refrigerated storage, or cooking. The technique consisted of an immunohistochemical detection of carbonyls by reaction with the specific probe DNPH (2,4-dinitrophenylhydrazine) followed by the sequential addition of a first antibody against DNPH-carbonylated proteins and a CY3-labeled secondary antibody. The fluorescence of the CY3 probe increased regularly with level of free radical generating system and storage time. Moreover, an important heterogeneity of carbonyl distribution was observed, with a higher oxidation level at the periphery than inside the muscle cells. Cooking induced fluorescence increase only at the periphery of cells. Specific coloration of collagen by Sirius red showed that collagen was not involved in fluorescence. We can deduce that accumulation of oxidized proteins observed in the cell periphery was linked to membrane protein oxidation and not to connective tissue oxidation. Biochemical assays were performed in parallel on membrane and myofibrillar proteins to provide complementary quantitative data on level of oxidized proteins.     

Classification of gilthead sea bream (Sparus aurata) from 1H NMR lipid profiling combined with principal component and linear discriminant analysis

The combination of (1)H NMR fingerprinting of lipids from gilthead sea bream (Sparus aurata) with nonsupervised and supervised multivariate analysis was applied to differentiate wild and farmed fish and to classify farmed specimen according to their areas of production belonging to the Mediterranean basin. Principal component analysis (PCA) applied on processed (1)H NMR profiles made a clear distinction between wild and farmed samples. Linear discriminant analysis (LDA) allowed classification of samples according to the geographic origin, as well as for the wild and farmed status using both PCA scores and NMR data as variables. Variable selection for LDA was achieved with forward selection (stepwise) with a predefined 5% error level. The methods allowed the classification of 100% of the samples according to their wild and farmed status and 85-97% to geographic origin. Probabilistic neural network (PNN) analyses provided complementary means for the successful discrimination among classes investigated.     

Carbon-14 biolabeling of (+)-catechin and proanthocyanidin oligomers in willow tree cuttings.

Proanthocyanidin polymers, oligomers, and the structurally related monomer (+)-catechin were labeled by incorporation of radioactive precursors in shoots of willow tree (Salix caprea L.). [1-(14)C]-Acetate and [U-(14)C]-phenylalanine precursors were fed through the cut stems or petioles of leaves. Optimization of several parameters such as the nature and origin of the plant material, leaf maturity, nature, and quantity of radioactive precursor applied and the duration of metabolism led to incorporation yields of 3.2% and to specific activities of 500 microCi/g. Detailed characterization of the products (polymerization degree, procyanidin/prodelphinidin ratio, specific activities) and purification by chromatography are reported. Some sugars bound to radiolabeled proanthocyanidin polymers were removed by enzymic treatment with a mixture of glycosidases. A radioactive purity close to 100% and specific activities suitable for bioavailability studies were obtained.     

1-methylcyclopropene increases storability and shelf life in climacteric and nonclimacteric plums

The effect of 1-methylcyclopropene (1-MCP) at three different doses (0.25, 0.50, and 0.75 microL L(-1)) on the ripening processes of a climacteric, cv. Santa Rosa, and a suppressed climacteric type, cv. Golden Japan, plum was studied. For both cultivars, positive effects were observed in terms of inhibition of ethylene production and delays of the physical, chemical, and biochemical changes associated with ripening. 1-MCP-treated plums were firmer with lower weight loss, reduced degrees Brix/titratable acidity ratios, and lower color changes during cold storage and subsequent shelf life at 20 degrees C than controls. For most factors, the effectiveness of 1-MCP was dose-dependent in Santa Rosa but dose-independent for Golden Japan.     

Effects of tetracycline administration on the proteomic profile of pig muscle samples (L. dorsi)

Effect of tetracycline (TC) administration on the proteomic profile of pig muscle was evaluated by 2D electrophoresis and MALDI-TOF mass spectrometry. The TC content at slaughter was determined in L. dorsi samples by HPLC-DAD. Mean residual concentration of TC in the muscle of treated animals, calculated as the sum of TC and epi-TC was 126.3 microg/kg, indicating a rapid elimination of TC in this tissue. Several differential spots (n = 54, p < 0.05) were observed in protein profiles from control and treated animals. MALDI-TOF identification gave a positive match for 5 differential spots, that is, glycerol-3-phosphate dehydrogenase 1 (G3PD1), phosphoglycerate kinase 1, novelprotein (0610037L13Rik), leucine aminopeptidase 3 (LAP), and hypothetical protein isoform 2. Results show that proteomics could be a useful tool to reveal pharmacological treatments with TC, even if the possible uses of differential spots as biomarkers to detect illegal administration of TC require further studies. Different spot patterns as a consequence of TC treatments seem to be another interesting issue for the consequences on tissue metabolism and meat quality.     

Effects of continuous spectrum LEDs used in indoor cultivation of two coniferous species Pinus sylvestris L. and Abies borisii-regis Mattf

The effects of four different continuous spectrum LED light qualities on the growth characteristics of Pinus sylvestris L. (PS) and Abies borisii-regis Mattf. (AB) seedlings were studied. The seedlings were exposed for 35 days inside growth chambers to G2 (high in red and far-red), AP67 (high in blue and far-red), AP67-ARCH (high in green), and NS2 (high in blue including a small percentage in the UV area) LEDs, as well as fluorescent light (FL) as Control. Each species showed a unique light-adapted response. G2 treatment stimulated needle formation of PS seedlings, while AB seedlings were unaffected. Hypocotyl elongation was promoted by FL; however, LEDs resulted in compact plants with greater root development, especially under the AP67-ARCH and AP67 spectra for PS and AB, respectively. In PS, AP67-ARCH and G2 significantly increased dry weight, while AB was affected significantly by AP67 and NS2. Furthermore, root growth potential of both species was better after LED pre-cultivation than the FL. Therefore, the use of continuous spectrum LEDs can enhance desirable quality characteristics of seedlings, which may be advantageous in large scale seedling production for reforestation.     

Climate change will affect the ability of forest management to reduce gaps between current and presettlement forest composition in southeastern Canada

Landscape Ecology - Forest landscapes at the boreal–temperate ecotone have been extensively altered. Reducing the gap between current and presettlement forest conditions through...     

Agriculturally productive yet biodiverse: human benefits and conservation values along a forest-agriculture gradient in Southern Ethiopia

Landscape Ecology - It remains unclear how agricultural landscapes can best serve multiple purposes such as simultaneously maintaining agricultural productivity and conserving biodiversity. Our...     

<Emphasis Type="Italic">Lupinus albus</Emphasis> Conglutin Gamma Modifies the Gene Expressions of Enzymes Involved in Glucose Hepatic Production <Emphasis Type="Italic">In Vivo</Emphasis>

Lupinus albus seeds contain conglutin gamma (Cγ) protein, which exerts a hypoglycemic effect and positively modifies proteins involved in glucose homeostasis. Cγ could potentially be used to manage patients with impaired glucose metabolism, but there remains a need to evaluate its effects on hepatic glucose production. The present study aimed to analyze G6pc, Fbp1, and Pck1 gene expressions in two experimental animal models of impaired glucose metabolism. We also evaluated hepatic and renal tissue integrity following Cγ treatment. To generate an insulin resistance model, male Wistar rats were provided 30% sucrose solution ad libitum for 20 weeks. To generate a type 2 diabetes model (STZ), five-day-old rats were intraperitoneally injected with streptozotocin (150 mg/kg). Each animal model was randomized into three subgroups that received the following oral treatments daily for one week: 0.9% w/v NaCl (vehicle; IR-Ctrl and STZ-Ctrl); metformin 300 mg/kg (IR-Met and STZ-Met); and Cγ 150 mg/kg (IR-Cγ and STZ-Cγ). Biochemical parameters were assessed pre- and post-treatment using colorimetric or enzymatic methods. We also performed histological analysis of hepatic and renal tissue. G6pc, Fbp1, and Pck1 gene expressions were quantified using real-time PCR. No histological changes were observed in any group. Post-treatment G6pc gene expression was decreased in the IR-Cγ and STZ-Cγ groups. Post-treatment Fbp1 and Pck1 gene expressions were reduced in the IR-Cγ group but increased in STZ-Cγ animals. Overall, these findings suggest that Cγ is involved in reducing hepatic glucose production, mainly through G6pc inhibition in impaired glucose metabolism disorders.