RESUMO
Acute intermittent porphyria (AIP) is a heme pathway disorder caused by a decrease in the activity and synthesis of porphobilinogen deaminase. Thus, the first heme precursor 5-aminolevulinic acid (ALA) accumulates in the liver. Reactive oxygen species (ROS) resulting from ALA oxidation may be correlated to a higher incidence of hepatocellular carcinoma (HCC) in AIP patients. However, the molecular mechanisms of this relationship have not been thoroughly elucidated to date. In this study, we investigated the effect of increasing levels of ALA on the expression of proteins related to DNA repair, oxidative stress, apoptosis, proliferation and lipid metabolism. Primary rat hepatocytes were isolated by the collagenase perfusion method, lipoperoxidation was evaluated by a TBA fluorimetric assay and Western blotting was used to assess protein abundance. The data showed that ALA treatment promoted a dose-dependent increase of p53 expression, downregulation of Bcl-2, HMG-CoA reductase and OGG1 and an increase in lipoperoxidation. There was no alteration in the expression of the transcription factor NF-κB, catalase and superoxide dismutase. ALA oxidation products induced protein regulation patterns, suggesting the interconnection of cellular processes, such as the intrinsic pathway of apoptosis, redox homeostasis, cell proliferation, lipid metabolism and DNA repair. This study helps to elucidate the molecular mechanisms of hepatotoxicity mediated by ALA pro-oxidant effects and supports the hypothesis that ALA accumulation correlates with a higher incidence of hepatic carcinogenic events.
Assuntos
Ácido Aminolevulínico/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Carcinogênese/efeitos dos fármacos , Carcinoma Hepatocelular/metabolismo , Proliferação de Células/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Neoplasias Hepáticas/metabolismo , Masculino , Estresse Oxidativo/efeitos dos fármacos , Cultura Primária de Células , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio , Superóxido DismutaseRESUMO
We compared the application of high hydrostatic pressure (HHP) on unfrozen carpaccio (HHP at 20 °C) and on previously-frozen carpaccio (HHP at -30 °C). HHP at 20 °C changed the color. The pressure increase from 400 to 650 MPa and the time increment from 1 to 5 min at 400 MPa increased L* and b*. a* decreased only with 650 MPa for 5 min at 20 °C. The prior freezing of the carpaccio and the HHP at -30 °C minimized the effect of the HHP on the color and did not change the shear force, but increased expressible moisture as compared to the untreated carpaccio. HHP at 20 °C was more effective in reducing the counts of microorganisms (aerobic total count at 30 °C, Enterobacteriaceae, psychrotrophs viable at 6.5 °C and lactic acid bacteria) than HHP at -30 º C. With HHP at 20 °C, we observed a significant effect of pressure and time on the reduction of the counts.
Assuntos
Fast Foods/análise , Conservação de Alimentos/métodos , Qualidade dos Alimentos , Carne/análise , Animais , Argentina , Bovinos , Fenômenos Químicos , Temperatura Baixa , Dieta/etnologia , Enterobacteriaceae/crescimento & desenvolvimento , Enterobacteriaceae/isolamento & purificação , Fast Foods/microbiologia , Alimentos Congelados/análise , Alimentos Congelados/microbiologia , Pressão Hidrostática , Lactobacillales/crescimento & desenvolvimento , Lactobacillales/isolamento & purificação , Carne/microbiologia , Fenômenos Mecânicos , Viabilidade Microbiana , Pigmentação , Resistência ao Cisalhamento , Fatores de Tempo , Água/análiseRESUMO
Beef muscles submitted to four enhancement treatments (1.88% whey protein concentrate (WPC)+1.25% sodium chloride (NaCl); 1.88% modified whey protein concentrate (MWPC)+1.25%NaCl; 0.25% sodium tripolyphosphate (STPP)+1.25%NaCl; 1.25%NaCl) and a control treatment (non-injected muscles) were sous vide cooked. Muscles with STPP+NaCl presented a significantly higher total yield (106.5%) in comparison to those with WPC/MWPC+NaCl (94.7% and 92.9%, respectively), NaCl alone (84.8%) or controls (72.1%). Muscles with STPP+NaCl presented significantly lower shear force values than control ones; also, WPC/MWPC+NaCl added muscles presented similar values than those from the other treatments. After cooking, muscles with STPP+NaCl or WPC/MWPC+NaCl depicted compacted and uniform microstructures. Muscles with STPP+NaCl showed a pink colour, meanwhile other treatment muscles presented colours between pinkish-grey and grey-brown. STPP+NaCl added samples presented the highest values of global tenderness and juiciness. The addition of STPP+NaCl had a better performance than WPC/MWPC+NaCl. However, the addition of WPC/MWPC+NaCl improved total yield in comparison to NaCl added or control ones.
Assuntos
Culinária/métodos , Aditivos Alimentares/administração & dosagem , Carne , Proteínas do Leite/administração & dosagem , Músculo Esquelético/química , Animais , Bovinos , Fenômenos Químicos , Cor , Concentração de Íons de Hidrogênio , Polifosfatos/administração & dosagem , Cloreto de Sódio/administração & dosagem , Paladar , Proteínas do Soro do LeiteRESUMO
BACKGROUND: Keratinocyte life span is modulated by receptors that control proliferation and differentiation, key processes during cutaneous tissue repair. The kinin B(1) receptor (B(1)R) has been reported in normal and pathological human skin, but so far there is no information about its role in keratinocyte biology. OBJECTIVES: To determine the consequence of kinin B(1)R stimulation on tyrosine phosphorylation, a key signalling mechanism involved in keratinocyte proliferation and differentiation. METHODS: Subconfluent primary cultures of human keratinocytes were used to investigate tyrosine phosphorylation, epidermal growth factor receptor (EGFR) transactivation, cell proliferation and keratinocyte differentiation. Cell proliferation was assessed by measuring bromodeoxyuridine incorporation whereas assessment of cell differentiation was based on the expression of filaggrin, cytokeratin 10 (CK10) and involucrin. RESULTS: The major proteins phosphorylated, after B(1)R stimulation, were of molecular mass 170, 125, 89 and 70 kDa. The 170- and 125-kDa proteins were identified as EGFR and p125(FAK), respectively. Phosphorylation was greatly reduced by GF109203X and by overexposure of keratinocytes to phorbol 12-myristate 13-acetate, indicating the participation of protein kinase C. B(1)R stimulation did not increase [Ca(2+)]i, but triggered EGFR transactivation, an event that involved phosphorylation of Tyr(845), Tyr(992) and Tyr(1068) of EGFR. B(1)R stimulation did not elicit keratinocyte proliferation, but triggered cell differentiation, visualized as an increase of filaggrin, CK10 and involucrin. Blockade of EGFR tyrosine kinase by AG1478, before B(1)R stimulation, produced an additional increase in filaggrin expression. CONCLUSIONS: The kinin B(1)R may contribute to keratinocyte differentiation and migration by triggering specific tyrosine signalling pathways or by interacting with the ErbB receptor family.
Assuntos
Diferenciação Celular , Receptores ErbB/metabolismo , Queratinócitos/citologia , Cininas/metabolismo , Receptor B1 da Bradicinina/metabolismo , Receptor B2 da Bradicinina/metabolismo , Células Cultivadas/metabolismo , Proteínas Filagrinas , Humanos , Queratinócitos/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Pele/metabolismoRESUMO
Sodium chloride (NaCl, 0-1.4%) and sodium tripolyphosphate (STPP, 0-0.5%) were added to Semitendinosus muscles and submitted to sous vide cooking at different temperatures (55-75°C). The effects of these three factors on pH, cooking loss, instrumental colour parameters, protein solubilization and distribution, and micro- and ultra-structure were evaluated. Quadratic surface responses equations were obtained from data (pH, cooking loss and colour parameters) as a function of the salts concentrations and cooking temperature. Both salts - alone or in combination - successfully reduced cooking loss. The best results were obtained for the combinations 0.25%STPP+1.20%NaCl and 0.25%STPP+0.70%NaCl, and temperatures between 60 and 65°C. Under these conditions, cooking loss was reduced close to 0%. pH was only dependent on STPP concentration, with a threshold concentration value of 0.25%. Temperature increment and NaCl addition produced a redness reduction. STPP incorporation recovered partially this parameter in comparison to non-added samples. Microscopy and SDS-PAGE results support the effect of the selected combinations of factors, suggesting that both salts together induced protein solubilization and gelation upon heating.
RESUMO
Research was undertaken to investigate how the addition of sodium chloride (NaCl) and/or sodium tripolyphosphate (TPP) to sous vide cooked meat pieces produces an increase in water holding capacity (WHC). Semitendinosus muscles were injected to obtain tissue final concentrations of 0.70% NaCl, 0.25% TPP, 0.70% NaCl+0.25% TPP, and 1.20% NaCl+0.25% TPP. SDS-PAGE analysis showed increased protein solubilization in those treatments which included NaCl. Thermal analysis of whole muscles and isolated myofibrils showed the destabilizing effect of NaCl and a global stabilizing effect of TPP. Both salts together induced a destabilizing global effect, where TPP assisted NaCl in breaking the meat structure. It is suggested that the WHC increments are related to conformational changes in myofibrillar proteins and to the weakening of myofibrillar structure by the removal of myofibrillar proteins.
RESUMO
Response surface methodology was used to optimize the effect of cooking temperature (CT: 65-75°C) and the incorporation of whey protein concentrate (WPC: 0-3.5%) and sodium chloride (NaCl: 0-2.5%) on technological, physical and sensory characteristics of cooked whole-muscle beef. Post-injection weight loss diminished when NaCl concentration increased. Moreover, the increment of both additives produced a reduction of cooking loss. An opposite effect was observed with the increment of CT. As it was expected, a total yield improvement was achieved by increasing both ingredients and diminishing CT. Equivalent yields are achieved complementing both ingredients, meaning that if one ingredient concentration is reduced the other has to be increased. Shear force values were not affected by the studied factors. Instead, lightness was reduced by their increment. At 65°C, injected muscles had lower flavour and odour scores than control. At all CT analyzed, the incorporated brines improved juiciness and tenderness-related attributes. Present results recommend the use of a CT of 70°C and maxima WPC and NaCl concentrations of 2.6% and 1.9%, respectively.
RESUMO
Beef muscles cooked by the sous vide system were evaluated for the effects of pre-injection tumbling, brine addition and post-injection tumbling on technological parameters, physical properties, visual appearance and tissue microstructure. The muscles were injected at 120% (over original weight) with a brine formulated to give a concentration of 3.5% whey protein concentrate and 0.7% sodium chloride on an injected raw product basis. Pre-injection tumbling did not affect most of the evaluated parameters. Brine addition reduced significantly the cooking and total weight losses. Total weight loss was 7.2% for injected muscles, and significantly higher (28.2%) for non-injected ones. Brine incorporation increased pH and reduced shear force values of cooked muscles. Extended post-injection tumbling (5rpm-10h) improved brine distribution and visual appearance, and also diminished the shear force values of cooked muscles. However, this treatment increased the weight losses of post-injection tumbling and cooking-pasteurization stages.
RESUMO
Los problemas de salud más frecuentes fueron: Hipertensión arterial: 46 por ciento. Incontinencia urinaria: 30 por ciento. Enfermedad coronaria: 15 por ciento. Caídas: 15 por ciento. Obesidad: 12 por ciento. Problemas de salud no referidos por los pacientes: Incontinencia urinaria: 30 por ciento. Caídas: 37 por ciento. Constipación: 7 por ciento. Trastornos prostáticos: 7 por ciento. Elevada proporción de pacientes con Polofarmacia: Pudiendo ocasionar Caídas e Institucionalizaciones. La gran mayoría eran independientes para las actividades instrumentales de la vida diaria, dada su condición de ambulatorios. A partir de este relevamiento se sugieren nuevas líneas de trabajo: Corroborar la adherencia de los hipertensos a la medicación. Evaluar indicaciones de medicaciones y su utilidad terapéutica. Un elevado porcentaje de los pacientes evaluados no tenían médico de referencia. Actualmente continúan ingresando pacientes en el protocolo.