Evaluación de aceitunas negras estilo griego utilizando salmueras de diferentes concentraciones

hree fermentation processes with black table-olives were tested. Olives were placed in: a) 16 %(w/v) concentration of NaCl, (traditional treatment), b) a buffer of CH3COOH (0.05M) and Ca(OH)2, (0.025M) without any NaCl and initial pH 4.7, and c) a buffer of CH3COOH (0.05M) and Ca(OH)2 (0.025M) containing 12.8 % (w/v) NaCl, and pH 4.3. Isolation, identification and enumeration of predominant microorganisms from fruits and brines sampled during the fermentation periods as well as color, intensity, texture and sensory evaluation tests of the final products were conducted. The third fermentation process, (c), yielded a product with low salt content no presence of spoilage microflora or other alterations during the fermentation period, with significantly better final texture and color, and higher acceptability among the consumers (PSe han ensayado tres procesos para la elaboración de aceitunas negras de mesa. Los frutos se colocaron en: a) una salmuera con una concentración de sal del 16 % (w/w), proceso tradicional; b) en una solución tampón compuesta de CH3COOH (0.05M) y Ca(OH)2 (0.025M) , sin NaCl y con un pH inicial de 4.3; c) una solución tampón compuesta de CH3COOH (0.05M) y Ca(OH)2 (0.025M),conteniendo 12.8 % (w/w) NaCl y un pH de 4.3. Se realizó el aislamiento, identificación y el recuento de los micoorganismos predominantes, tanto del fruto como de las salmueras, durante la fermentación. Asimismo, se estudió el color, textura y las características organolépticas de los productos finales. El tercer tipo de proceso fermentativo, tipo c, dio lugar a un producto con baja sal, ausencia de microorganismos alterantes o de cualquier otra alteración, dando una textura y un color significativamente mejor y resultando con una mayor aceptación entre los consumidores (

Mitochondrial targeting adaptation of the hominoid-specific glutamate dehydrogenase driven by positive Darwinian selection

Many new gene copies emerged by gene duplication in hominoids, but little is known with respect to their functional evolution. Glutamate dehydrogenase (GLUD) is an enzyme central to the glutamate and energy metabolism of the cell. In addition to the single, GLUD-encoding gene present in all mammals (GLUD1), humans and apes acquired a second GLUD gene (GLUD2) through retroduplication of GLUD1, which codes for an enzyme with unique, potentially brain-adapted properties. Here we show that whereas the GLUD1 parental protein localizes to mitochondria and the cytoplasm, GLUD2 is specifically targeted to mitochondria. Using evolutionary analysis and resurrected ancestral protein variants, we demonstrate that the enhanced mitochondrial targeting specificity of GLUD2 is due to a single positively selected glutamic acid-to-lysine substitution, which was fixed in the N-terminal mitochondrial targeting sequence (MTS) of GLUD2 soon after the duplication event in the hominoid ancestor ~18–25 million years ago. This MTS substitution arose in parallel with two crucial adaptive amino acid changes in the enzyme and likely contributed to the functional adaptation of GLUD2 to the glutamate metabolism of the hominoid brain and other tissues. We suggest that rapid, selectively driven subcellular adaptation, as exemplified by GLUD2, represents a common route underlying the emergence of new gene functions

A methodological approach for optimum preservation results: The packaging paradigm

The food preservation hypothesis as impacted by overall packaging applications is considered in this work. The objective was to devise a decision supportive method for the selection of “just-right” packaging materials, techniques and procedures. For that, food preservation was critically approached in order to identify the optimum outcome at experimental and packaging selection decision-making levels. A mathematically supported and proven knowledge classification, and the establishment of a straightforward coherence mode among the principles of the natural systemic phenomena, were used. The ultimate aim of this work was to justifiably surpass a simple description of packaging according to its measurable specifications, and instead, engage its inherent properties into a cyclic 8-steps-process for eventually understanding its potential to support any particular preservation hypothesis in question. The proposed methodology includes primarily, the consideration of the study hypothesis and, in parallel, the conclusive remarks and claims with respect to the experimental factors involved (properties, parameters, relations and conditions). Considering the experimentally controlled set-ups that a researcher has to expose the food system to and the role of packaging in obtaining its preservation potential, our method supports the experimenters in selecting the experimental conditions under which the preservation hypothesis can be disclaimed and furthermore, it could indicate the way to reduce experimentation research waste. </p

Human GLUD2 Glutamate Dehydrogenase Is Expressed in Neural and Testicular Supporting Cells*

Mammalian glutamate dehydrogenase (GDH) is an allosterically regulated enzyme that is expressed widely. Its activity is potently inhibited by GTP and thought to be controlled by the need of the cell for ATP. In addition to this housekeeping human (h) GDH1, humans have acquired (via a duplication event) a highly homologous isoenzyme (hGDH2) that is resistant to GTP. Although transcripts of GLUD2, the gene encoding hGDH2, have been detected in human neural and testicular tissues, data on the endogenous protein are lacking. Here, we developed an antibody specific for hGDH2 and used it to study human tissues. Western blot analyses revealed, to our surprise, that endogenous hGDH2 is more densely expressed in testis than in brain. At the subcellular level, hGDH2 localized to mitochondria. Study of testicular tissue using immunocytochemical and immunofluorescence methods revealed that the Sertoli cells were strongly labeled by our anti-hGDH2 antibody. In human cerebral cortex, a robust labeling of astrocytes was detected, with neurons showing faint hGDH2 immunoreactivity. Astrocytes and Sertoli cells are known to support neurons and germ cells, respectively, providing them with lactate that largely derives from the tricarboxylic acid cycle via conversion of glutamate to α-ketoglutarate (GDH reaction). As hGDH2 is not subject to GTP control, the enzyme is able to metabolize glutamate even when the tricarboxylic acid cycle generates GTP amounts sufficient to inactivate the housekeeping hGDH1 protein. Hence, the selective expression of hGDH2 by astrocytes and Sertoli cells may provide a significant biological advantage by facilitating metabolic recycling processes essential to the supportive role of these cells

Gain-of-function variant in GLUD2 glutamate dehydrogenase modifies Parkinson's disease onset

Parkinson's disease (PD), a common neurodegenerative disorder characterized by progressive loss of dopaminergic neurons and their terminations in the basal ganglia, is thought to be related to genetic and environmental factors. Although the pathophysiology of PD neurodegeneration remains unclear, protein misfolding, mitochondrial abnormalities, glutamate dysfunction and/or oxidative stress have been implicated. In this study, we report that a rare T1492G variant in GLUD2, an X-linked gene encoding a glutamate dehydrogenase (a mitochondrial enzyme central to glutamate metabolism) that is expressed in brain (hGDH2), interacted significantly with age at PD onset in Caucasian populations. Individuals hemizygous for this GLUD2 coding change that results in substitution of Ala for Ser445 in the regulatory domain of hGDH2 developed PD 6–13 years earlier than did subjects with other genotypes in two independent Greek PD groups and one North American PD cohort. However, this effect was not present in female PD patients who were heterozygous for the DNA change. The variant enzyme, obtained by substitution of Ala for Ser445, showed an enhanced basal activity that was resistant to GTP inhibition but markedly sensitive to modification by estrogens. Thus, a gain-of-function rare polymorphism in hGDH2 hastens the onset of PD in hemizygous subjects, probably by damaging nigral cells through enhanced glutamate oxidative dehydrogenation. The lack of effect in female heterozygous PD patients could be related to a modification of the overactive variant enzyme by estrogens