RESUMEN
Italian salami were sonicated in different times (0, 3, 6 and 9min) using ultrasound bath (US, 25kHz). The effect of sonication on microbial growth (lactic acid bacteria and Micrococcaceae), lipid and protein oxidation, total heme pigments (THP), non heme iron (NHI) and metmyoglobin (MMb) was investigated during processing (0, 2, 15, and 28days) and storage (1, 30, and 120days). US enhanced growth of microorganisms (P<0.05), mainly for the treatment 9min of sonication. The lipid (peroxide value and TBARS) and protein (thiol group) oxidative reactions were accelerated by US (P<0.05) and they should be considered to maintain Italian salami quality. Sonication contributed to maintenance of THP (P<0.05), especially during storage. MMb pigment was not affected by sonication (P>0.05). This study presented some features of US application that could be explored in the manufacture of Italian salami.
Asunto(s)
Fenómenos Químicos/efectos de la radiación , Microbiología de Alimentos , Productos de la Carne/efectos de la radiación , Sonicación , Fermentación , Manipulación de Alimentos/métodos , Hemo/análisis , Hemo/efectos de la radiación , Hierro/análisis , Hierro/efectos de la radiación , Italia , Lactobacillales/crecimiento & desarrollo , Lactobacillales/efectos de la radiación , Peroxidación de Lípido/efectos de la radiación , Productos de la Carne/análisis , Productos de la Carne/microbiología , Metamioglobina/análisis , Metamioglobina/efectos de la radiación , Micrococcaceae/crecimiento & desarrollo , Micrococcaceae/efectos de la radiación , Oxidación-Reducción/efectos de la radiación , Proteínas/química , Proteínas/efectos de la radiación , Sustancias Reactivas al Ácido Tiobarbitúrico/análisisRESUMEN
The aim of the present work was to evaluate the redox and oligomeric effects associated with the human hemoglobin of stored red blood cells that had been previously submitted to gamma radiation. Whole blood was collected from healthy donors and irradiated with 25 Gy of γ-radiation within 24 h of collection. At days 3, 5, 7, 9, 11, 14, and 28 postirradiation, fractions were removed and centrifuged, and the levels of methehemoglobin and oxyhemoglobin were measured. Hb was isolated to measure the denaturation and UV-vis spectra. The results from electrophoresis demonstrated that there was no fragmentation or cross-linking of the hemoglobin. However, ferrous center oxidation was identified as a very significant process. This mechanism is likely through an autoxidation process of the ferrous heme center, which has a maximal intensity between 5 and 7 days of storage. Interestingly, a subsequent reduction of the oxidized heme species was observed, and after 9 days of storage, the difference between the ferric species present in the control and irradiated samples was not representative. This interesting fact suggests a type of "protective action" by the blood to control the oxidative stress generated by the gamma irradiation. The UV-vis measurements demonstrated that the oxidized species was predominantly formed by hemichrome species (bis-histidine ferric heme species), which are usually associated with Heinz bodies. After 28 days of storage, evidence from the UV-vis measurements indicated that the oxidation of the irradiated sample was much higher than that observed in the control sample. These results demonstrate that despite the minimal polypeptide changes observed in the hemoglobin of stored red blood cells after gamma irradiation, the oxidation of the heme metallic center is not irrelevant and must be controlled to improve the hematological clinical procedures associated with the storage of red blood cells.
Asunto(s)
Conservación de la Sangre , Eritrocitos/efectos de la radiación , Rayos gamma/efectos adversos , Hemoglobinas/efectos de la radiación , Procedimientos de Reducción del Leucocitos/métodos , Electroforesis de las Proteínas Sanguíneas , Hemo/efectos de la radiación , Hemoglobinas/ultraestructura , Humanos , Metahemoglobina/análisis , Oxidación-Reducción , Estrés Oxidativo , Oxihemoglobinas/análisis , Conformación Proteica/efectos de la radiación , Desnaturalización Proteica , Sulfahemoglobina/análisis , Factores de TiempoRESUMEN
N-dodecyl-N'-(2-phosphonoethyl)-1,4,5,8-naphthalenetetracarboxylic diimide (DNDI) is a novel naphthalenic diimide with amphiphilic character. DNDI was synthesized through the sequential reaction of 1,4,5,8-naphthalenetetracarboxylic dianhydride, first with dodecylamine and then with 2-aminoethylphosphonic acid. Fluorescence measurements showed that DNDI forms excimers in water at sufficiently high concentrations. The fluorescence quantum yield of DNDI in diluted solutions is sensitive to the polarity of the microenvironment, decreasing as going from water to less polar solvents. This property allowed to monitor the incorporation of DNDI into cetyl trimethyl ammonium bromide (CTAB) micelles, with a binding constant of 1.2x10(4) M-1. UV irradiation (365 nm) of solutions containing DNDI and the redox protein cytochrome c (cyt c) resulted in the reduction of the heme iron from the Fe(III) to the Fe(II) state, a reaction that was inhibited by the incorporation of DNDI into CTAB micelles. DNDI formed host-guest complexes with alpha-cyclodextrin (alpha-CD) through the inclusion of the dodecyl group, resulting in an increased aqueous solubility of the compound.