RESUMO
Quality and safety of fresh produce are important to public health and maintaining commerce between Mexico and USA. While preventive practices can reduce risks of contamination and are generally successful, the variable environment of the supply chain of fresh produce can be suitable for introduction or proliferation of pathogenic microorganisms. As routine surveillance of these pathogens is not practical, indicator microorganisms are used to assess the sanitary conditions of production and handling environments. An opportunity exists to use indicators on fresh produce to measure how handling and transport from field to market may affect microbial populations that contribute to their quality or safety. The objective was to quantify indicator microorganisms on tomatoes sampled along the supply chain during the harvest year, in order to observe the levels and changes of populations at different locations. Roma tomatoes (n=475) were taken from the same lots (n=28) at four locations of the postharvest supply chain over five months: at arrival to and departure from the packinghouse in México, at the distribution center in Texas, and at retail in USA. Samples were analyzed individually for four microbial populations: aerobic plate count (APC), total coliforms (TC), generic Escherichia coli, and yeasts and molds (YM). APC population differed (p<0.05) from 1.9±1.1, 1.7±1.1, 2.3±1.1 and 3.5±1.4logCFU/g at postharvest, packing, distribution center and supermarket, respectively. TC populations were <1logCFU/g at postharvest, increased at packing (0.7±1.0logCFU/g), decreased in distribution (0.4±0.8logCFU/g) and increased in supermarkets (1.4±1.5logCFU/g). Generic E. coli was not identified from coliform populations in this supply chain. YM populations remained <1logCFU/g, with the exception of 1.1±1.3logCFU/g at supermarkets and tomatoes were not visibly spoiled. The levels reported from this pilot study demonstrated the dynamics within populations as influenced by time and conditions in one supply chain during a harvest year, while the large variances in some locations indicate opportunities for improvement. Overall, packinghouse and supermarket locations were identified as crucial points to control microbial safety risks.
Assuntos
Bactérias/isolamento & purificação , Contaminação de Alimentos/análise , Frutas/microbiologia , Solanum lycopersicum/microbiologia , Bactérias/classificação , Bactérias/genética , Contagem de Colônia Microbiana , Contaminação de Alimentos/economia , Microbiologia de Alimentos/economia , Frutas/economia , Humanos , México , Projetos PilotoRESUMO
Recent studies have shown that the epithelial sodium channel (ENaC) is expressed in vascular tissue. However, the role that ENaC may play in the responses to vasoconstrictors and NO production has yet to be addressed. In this study, the contractile responses of perfused pressurized small-diameter rat mesenteric arteries to phenylephrine and serotonin were reduced by ENaC blockade with amiloride (75.1+/-3.2% and 16.9+/-2.3% of control values, respectively; P<0.01) that was dose dependent (EC(50)=88.9+/-1.6 nmol/L). Incubation with benzamil, another ENaC blocker, had similar effects. alpha, beta, and gamma ENaC were identified in small-diameter rat mesenteric arteries using RT-PCR and Western blot with specific antibodies. In situ hybridization and immunohistochemistry localized ENaC expression to the tunica media and endothelium of small-diameter rat mesenteric arteries. Patch-clamp experiments demonstrated that primary cultures of mesenteric artery endothelial cells expressed amiloride-sensitive sodium currents. Mechanical ablation of the endothelium or inhibition of eNOS with N(omega)-nitro-L-arginine inhibited the reduction in contractility caused by ENaC blockers. ENaC inhibitors increased eNOS phosphorylation (Ser 1177) and Akt phosphorylation (Ser 473). The presence of the phosphoinositide 3-kinase inhibitor LY294002 blunted Akt phosphorylation and eNOS phosphorylation and the decrease in the response to phenylephrine caused by blockers of ENaC, indicating that the phosphoinositide 3-kinase/Akt pathway was activated after ENaC inhibition. Finally, we observed that the effects of blockers of ENaC were flow dependent and that the vasodilatory response to shear stress was enhanced by ENaC blockade. Our results identify a previously unappreciated role for ENaC as a negative modulator of eNOS and NO production in resistance arteries.
Assuntos
Endotélio Vascular/fisiologia , Canais Epiteliais de Sódio/metabolismo , Artérias Mesentéricas/fisiologia , Óxido Nítrico Sintase/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Amilorida/análogos & derivados , Amilorida/farmacologia , Análise de Variância , Animais , Velocidade do Fluxo Sanguíneo , Western Blotting , Endotélio Vascular/efeitos dos fármacos , Canais Epiteliais de Sódio/efeitos dos fármacos , Imuno-Histoquímica , Hibridização In Situ , Masculino , Artérias Mesentéricas/efeitos dos fármacos , Modelos Animais , Óxido Nítrico/metabolismo , Fosforilação , Probabilidade , Distribuição Aleatória , Ratos , Ratos Sprague-DawleyRESUMO
We have investigated the relationship between the status of red blood cells (RBCs) and their susceptibility to toxin sticholysin II (StII) hemolytic activity; we have evaluated this effect in different RBC ensembles, comprising young and old cells, and in cells partially damaged by their pre-exposition to a free radical source. Upon action of StII, young cell populations are less prone to hemolysis than the whole population, while old cell populations and peroxyl-oxidized red cells are lysed faster than the whole population. Cell K(+) content was higher in young cells and lower in both senescent cells and in peroxyl-damaged cells relative to whole cell population. The relevance of cell K(+) content in St II-induced lysis was shown when external Na(+) was partially replaced by K(+); under this condition, RBC lysed faster in the presence of St II but no difference was observed among young cells, whole cells population and peroxyl-damaged cells; only old cells lysed faster that the whole population, response that can be due to an enhanced St II-induced pore formation as supported by evaluation of St II irreversible binding to RBC. It is concluded that this factor and the amount of intracellular K(+) are the dominant parameters that modulate the resistance of RBC to St II-induced lysis.