Erk1/2 signaling mediates the HGF-induced protection against ethanol and acetaldehyde-induced toxicity in the pancreatic RINm5F cell line

Alcohol-induced pancreas damage remains as one of the main risk factors for pancreatitis development. This disorder is poorly understood, particularly the effect of acetaldehyde, the primary alcohol metabolite, in the endocrine pancreas. Hepatocyte growth factor (HGF) is a protective protein in many tissues, displaying antioxidant, antiapoptotic, and proliferative responses. In the present work, we were focused on characterizing the response induced by HGF and its protective mechanism in the RINm5F pancreatic cell line treated with ethanol and acetaldehyde. RINm5F cells were treated with ethanol or acetaldehyde for 12 h in the presence or not of HGF (50 ng/ml). Cells under HGF treatment decreased the content of reactive oxygen species and lipid peroxidation induced by both toxics, improving cell viability. This effect was correlated to an improvement in insulin expression impaired by ethanol and acetaldehyde. Using a specific inhibitor of Erk1/2 abrogated the effects elicited by the growth factor. In conclusion, the work provides mechanistic evidence of the HGF-induced-protective response to the alcohol-induced damage in the main cellular component of the endocrine pancreas.     

Branched chain amino acids chronic treatment and muscular exercise performance in athletes: a study through plasma acetyl-carnitine levels

Summary In relation to energy request during physical exercise, muscular tissue Branched Chain Amino Acids (BCAA) are metabolized particularly when the oxidation rises. But in the whole-human body, it is difficult to estimate, in a quantitative sense, the role played by BCAA in sustaining exercise. During a BCAA treatment, made on a group of athletes kept under observation, it was observed, through Conconi's test, that this treatment influenced physical performance. Aim of present work is to investigate if BCAA chronic treatment effect on physiological trial is confirmed on blood circulating biochemical energy parameters and in particular on acetyl-carnitine, since acetyl-linked compounds may be an important biochemical factor.Fourteen athletic well trained male subjects, were randomly divided into two subgroups; a first group was submitted to a chronic treatment (n = 7) of BCAA (oral intake was 0.2 g/Kg die) and a second group, as controls (n = 7), assumed oral placebo. Conconi's test demonstrated a significant difference (p < 0.005) in the exercise performance of the two sub-groups, comparing the measurements of ratios of deflection velocity (V _d ), before and after the treatment. Therefore we studied the athletes performing a muscular exercise test (40 Km/h, cycle race, for 90 min) after one month of treatment. During this treatment period the subjects followed a well standardized diet. Samples of blood were drawn before, at the end and during the recovery (60 min) to study if traditional biochemical parameters varied and confirmed the observed differences in Conconi's test. The measurements of concentrations of FFA, KB, free carnitine, acetyl-carnitine and BCAA were performed. Plasma BCAA levels did not demonstrate variations either before or after the exercise performance, or between the two groups. The biochemical factors, substrates and hormones, KB, FFA, lactate, insulin and growth hormone plasma levels did not demonstrate significant differences from the patterns present in literature. Plasma free and acetyl-carnitine followed the well known variations, but only acetyl-carnitine levels demonstrated, at the end increase in acetyl-carnitine levels could be related to a minor fatigue situation and to a larger energy supply availability perhaps present in BCAA treated athletes (Sahlin et al., 1990; May et al., 1989). Both mentioned hypothesis seem in concordance with a smaller acetyl-CoA substrate accumulation, or better for present study, is even more successful with athletes who give a better physical performance. In fact Conconi's test in the two sub-groups of athletes seems to suggest that BCAA treated athletes were able to give a better performance, furthermore out of curiosity we point out that the athletes treated with BCAA won more races than the untreated.We would also like to add in conclusion that although confirming the difficulties of studies in the whole-body, our work gives an interesting clue about the possibility to use acetyl-carnitine plasma levels to understand the biochemical importance of the BCAA as substrate able to influence physical performance, but further research is needed.The phenomenon presence might be showed better perhaps by studying untrained groups during prolonged exercise and with physical performance at exhaustion. If treatment were able to help the physical performance and to shift the fatigue, then confirmation might be a less raised plasma acetyl-carnitine level. In effect blood ammonium levels in present study did not demonstrate any variation in and between sub-groups; this latter observation could be caused by the quantity of work load, and training state of the athletes (Ji et al., 1987; Kirkendall, 1990). Moreover, as observed by Hageloch et al. (1990), the ammonia increases less during prolonged endurance exercise, and in fact the athletes of present study were all middle distance racing cyclists, and the physical performance was a prolonged endurance exercise.     

Copper Toxicity in Maize: The Severity of the Stress is Reduced Depending on the Applied Fe-Chelating Agent

Journal of Plant Growth Regulation - The wide use of copper (Cu)-based fungicide has caused a stepwise accumulation of Cu in the environment increasing the occurrence of phytotoxicity in crops. To...     

Molecular characterization of the proteinase-encoding gene, prb1, related to mycoparasitism by Trichoderma harzianum

The soil fungus Trichoderma harzianum is a mycoparasitic fungus known for its use as a biocontrol agent of phytopathogenic fungi. Among other factors, Trichoderma produces a series of antibiotics and fungal cell wall-degrading enzymes. These enzymes are believed to play an important role in mycoparasitism. Among the hydrolytic enzymes, we have identified a basic proteinase (Prb1) which is induced by either autoclaved mycelia, fungal cell wall preparation or chitin; however, the induction does not occur in the presence of glucose. The proteinase was purified and biochemically characterized as a serine proteinase of 31 kDa and pl 9.2. Based on the sequence of three internal peptides, synthetic oligonudeotide probes were designed. These probes allowed subsequent isolation of a cDNA and its corresponding genomic clone. The deduced amino acid sequence indicates that the proteinase is synthesized as a pre-proenzyme and allows its classification as a serine proteinase. Northen analysis shows that the induction of this enzyme is due to an increase in the corresponding mRNA level.     

Phytochemical Characterization of Cannabis sativa L. Roots from Northeastern Brazil

This article describes the phytochemical study of Cannabis sativa roots from northeastern Brazil. The dried plant material was pulverized and subjected to exhaustive maceration with ethanol at room temperature, obtaining the crude ethanolic extract (Cs-EEBR). The volatile compounds were analyzed by gas chromatography coupled with mass spectrometry (GC/MS), which allowed to identify 22 compounds by comparing the linear retention index (LRI), the similarity index (SI) and the fragmentation pattern of the constituents with the literature. By this technique the major compounds identified were: friedelan-3-one and β-sitosterol. In addition, two fractions were obtained from Cs-EEBR by classical column chromatography and preparative thin layer chromatography. These fractions were analyzed by NMR and IR and together with the mass spectrometry data allowed to identify the compounds: epifriedelanol, friedelan-3-one, β-sitosterol and stigmasterol. The study contributed to the phytochemical knowledge of Cannabis sativa, specifically the roots, as there are few reports on the chemical constituents of this part of the plant.     

BLOEM: A spatially explicit model of bioenergy and carbon capture and storage,applied to Brazil

Bioenergy could play a major role in decarbonizing energy systems in the context of the Paris Agreement. Large-scale bioenergy deployment could be related to sustainability issues and requires major infrastructure investments. It, therefore, needs to be studied carefully. The Bioenergy and Land Optimization Spatially Explicit Model (BLOEM) presented here allows for assessing different bioenergy pathways while encompassing various dimensions that influence their optimal deployment. In this study, BLOEM was applied to the Brazilian context by coupling it with the Brazilian Land Use and Energy Systems (BLUES) model. This allowed investigating the most cost-effective ways of attending future bioenergy supply projections and studying the role of recovered degraded pasture lands in improving land availability in a sustainable and competitive manner. The results show optimizing for limiting deforestation and minimizing logistics costs results in different outcomes. It also indicates that recovering degraded pasture lands is attractive from both logistics and climate perspectives. The systemic approach of BLOEM provides spatial results, highlighting the trade-offs between crop allocation, land use and the logistics dynamics between production, conversion, and demand, providing valuable insights for regional and national climate policy design. This makes it a useful tool for mapping sustainable bioenergy value chain pathways.     

Litter quality and nutrient cycling affected by grazing-induced species replacements along a precipitation gradient

One of the potential mechanisms for the impact of herbivores on nutrient cycling is the effect of selective grazing on litter quality through changes in species composition. However, the scarce evidence collected on this mechanism is controversial and seemingly influenced by site-specific variables. In this paper, we explored the consequences of grazing-induced changes in species composition on litter quality and nitrogen cycling with a regional perspective. Along a 900-mm of mean annual rainfall gradient, we selected species promoted and diminished by grazing from three natural rangelands of Argentina, analyzed their litter quality, and determined their decomposition and nutrient release kinetics under common greenhouse conditions. Litter quality and decomposition rates were strongly associated with plant response to grazing. However, the magnitude and direction of these differences depended on the ecosystem considered. In the wettest site, the species promoted by grazing (forbs) had higher nitrogen and phosphorus contents, faster decomposition rates, and higher release of nitrogen to the soil than species diminished by grazing (C₃ and C₄ grasses). In the intermediate and dry sites, species promoted by grazing had lower nitrogen and phosphorus contents, and slower decomposition rates than those diminished by grazing (C₃ grasses in both cases). Decomposition of the entire group of species was not correlated with mean annual rainfall, but when litter of the species diminished by grazing was analyzed, it was negatively correlated with precipitation. Nitrogen was immobilized more often than mineralized, even after one year of incubation. Immobilization was negatively correlated with precipitation. All these results indicate that grazing may significantly alter nutrient cycling by affecting litter quality through changes in species composition. These effects seem to be larger when species replacements induced by grazing either involve functional groups, as it was the case in our wettest site, or change root to shoot ratios. Therefore, the functional groups involved in the replacement of species as well as shifts between belowground and aboveground allocation should play a key role in grazing-induced changes on nitrogen cycling.     

Electron paramagnetic resonance studies and effects of vanadium in Saccharomyces cerevisiae

Vanadium uptake by whole cells and isolated cell walls of the yeast Saccharomyces cerevisiae was studied. When orthovanadate was added to wild-type S. cerevisiae cells growing in rich medium, growth was inhibited as a function of the VO₄ ^3- concentration and the growth was completely arrested at a concentration of 20 mM of VO₄ ^3- in YEPD. Electron paramagnetic resonance (EPR) spectroscopy was used to obtain structural and dynamic information about the cell-associated paramagnetic vanadyl ion. The presence of EPR signals indicated that vanadate was reduced by whole cells to the vanadyl ion. On the contrary, no EPR signals were detected after interaction of vanadate with isolated cell walls. A mobile and an immobile species associated in cells with small chelates and with macromolecular sites, respectively, were identified. The value of rotational correlation time _r indicated the relative motional freedom at the macromolecular site. A strongly immobilized vanadyl species bound to polar sites mainly through coulombic attractions was detected after interaction of VO²⁺ ions with isolated cell walls.     

Single-cell investigation by laser scanning confocal microscopy of cytochemical alterations resulting from extracellular oxidant challenge

Confocal laser scanning fluorescence microscopy coupled to image analysis was employed in order to develop and evaluate procedures for the appraisal at the single-cell level of: (1) protein-bound 4-hydroxynonenal, the specific product of membrane peroxidation (by means of immunocytochemistry with biotin-avidin revelation); (2) protein oxidation (by reaction of protein carbonyls with 2,4-dinitrophenyl-hydrazine followed by immunocytochemistry of dinitrophenyl moieties); and (3) cellular protein thiols (by direct alkylation of sulfhydryl groups with thiol-specific fluorescent reagents possessing different cell permeabilities). The procedures proved able to reveal the subcellular distribution of cytochemical parameters useful as indices of oxidative stress conditions, and may allow redox phenotyping of isolated cells, which would provide an efficient tool in selected experimental models.