RESUMEN
This paper provides insights in the isothermal gelation behavior of white and red chicken myofibrillar proteins (CMP) at different temperatures (20 to 80°C) and the underlying aggregation mechanism, allowing understanding of structure formation in poultry products during thermal processing. At low temperatures (20 to 60°C), an increase in aromatic surface hydrophobicity (SoANS) was found, suggesting potential formation of hydrophobic interactions between CMP. At higher temperatures (60 to 80°C), high SoANS and a significant decrease in total sulfhydryl amount (SH-amount) strongly indicate the presence of hydrophobic interactions and disulfide bonding, resulting in aggregation, as suggested by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The elastic modulus G' after 60 min isothermal heating (G'60min) significantly increased at 70 or 80°C, depending on the type of CMP. Differences in G'60min between white and red CMP were rather small at low temperatures (20 to 60°C). However, at 70°C, white CMP reached higher G'60min compared to red CMP, while the opposite was observed at 80°C. Overall, for every temperature studied, SoANS and SH-amount of red CMP were higher compared to white CMP. The differences in G'60min, SoANS, and SH-amount between white and red CMP were probably due to different isoforms. The results may help to steer quality characteristics of poultry products through intelligent choice of processing conditions.
Asunto(s)
Electroforesis en Gel de Poliacrilamida/veterinaria , Geles/análisis , Carne/análisis , Proteínas Musculares/química , Miofibrillas/clasificación , TemperaturaRESUMEN
Equine herpesvirus type 1 (EHV-1) replicates extensively in the epithelium of the upper respiratory tract, after which it can spread throughout the body via a cell-associated viremia in mononuclear leukocytes reaching the pregnant uterus and central nervous system. In a previous study, we were able to mimic the in vivo situation in an in vitro respiratory mucosal explant system. A plaquewise spread of EHV-1 was observed in the epithelial cells, whereas in the connective tissue below the basement membrane (BM), EHV-1-infected mononuclear leukocytes were noticed. Equine herpesvirus type 4 (EHV-4), a close relative of EHV-1, can also cause mild respiratory disease, but a cell-associated viremia in leukocytes is scarce and secondary symptoms are rarely observed. Based on this striking difference in pathogenicity, we aimed to evaluate how EHV-4 behaves in equine mucosal explants. Upon inoculation of equine mucosal explants with the EHV-4 strains VLS 829, EQ(1) 012 and V01-3-13, replication of EHV-4 in epithelial cells was evidenced by the presence of viral plaques in the epithelium. Interestingly, EHV-4-infected mononuclear leukocytes in the connective tissue below the BM were extremely rare and were only present for one of the three strains. The inefficient capacity of EHV-4 to infect mononuclear cells explains in part the rarity of EHV-4-induced viremia, and subsequently, the rarity of EHV-4-induced abortion or EHM.
Asunto(s)
Infecciones por Herpesviridae/veterinaria , Herpesvirus Équido 1/patogenicidad , Herpesvirus Équido 4/patogenicidad , Enfermedades de los Caballos/virología , Caballos/virología , Animales , Células Epiteliales/virología , Infecciones por Herpesviridae/virología , Herpesvirus Équido 1/fisiología , Herpesvirus Équido 4/fisiología , Enfermedades de los Caballos/diagnóstico , Leucocitos Mononucleares/virología , Mucosa Nasal/virología , Técnicas de Cultivo de Tejidos , Ensayo de Placa Viral/veterinaria , Tropismo Viral , Viremia/veterinaria , Viremia/virología , Replicación ViralRESUMEN
Equine herpesvirus type 1 (EHV-1) is the causative agent of equine herpes myeloencephalopathy, of which outbreaks are reported with increasing frequency throughout North America and Europe. This has resulted in its classification as a potentially emerging disease by the US Department of Agriculture. Recently, it was found that a single nucleotide polymorphism (SNP) in the viral DNA polymerase gene (ORF30) at aa 752 (N-->D) is associated with the neurovirulent potential of EHV-1. In the present study, equine respiratory mucosal explants were inoculated with several Belgian isolates typed in their ORF30 as D(752) or N(752), to evaluate a possible difference in replication in the upper respiratory tract. In addition, to evaluate whether any observed differences could be attributed to the SNP associated with neurovirulence, the experiments were repeated with parental Ab4 (reference neurovirulent strain), parental NY03 (reference non-neurovirulent strain) and their N/D revertant recombinant viruses. The salient findings were that EHV-1 spreads plaquewise in the epithelium, but plaques never cross the basement membrane (BM). However, single EHV-1-infected cells could be observed below the BM at 36 h post-inoculation (p.i.) for all N(752) isolates and at 24 h p.i. for all D(752) isolates, and were identified as monocytic cells and T lymphocytes. Interestingly, the number of infected cells was two to five times higher for D(752) isolates compared with N(752) isolates at every time point analysed. Finally, this study showed that equine respiratory explants are a valuable and reproducible model to study EHV-1 neurovirulence in vitro, thereby reducing the need for horses as experimental animals.
Asunto(s)
ADN Polimerasa Dirigida por ADN/genética , Herpesvirus Équido 1/crecimiento & desarrollo , Herpesvirus Équido 1/patogenicidad , Caballos/virología , Mucosa Nasal/virología , Factores de Virulencia/genética , Replicación Viral , Animales , Bélgica , ADN Polimerasa Dirigida por ADN/fisiología , Herpesvirus Équido 1/aislamiento & purificación , Monocitos/virología , Mutación Missense , Técnicas de Cultivo de Órganos , Linfocitos T/virología , Factores de Tiempo , Proteínas Virales/genética , Proteínas Virales/fisiología , Virulencia , Factores de Virulencia/fisiologíaRESUMEN
Conformational alterations induced by streptomycin in the bacterial ribosome have been investigated using as probes, ethidium bromide, N-[14C]ethylmaleimide and a spin label nitroxide analog of N-ethylmaleimide. 1. The binding of the antibiotic to the ribosome does not affect the reactivity of sulfhydryl groups towards N-ethylmaleimide. 2. The motional freedom of spin labels bound to ribosomal proteins S1 and S18 is increased but it is hardly affected at other labeled sites. This observation suggests that the binding of streptomycin causes a local loosening of the ribosomal structure. 3. Ribosomes are found to bind less ethidium bromide in the presence of streptomycin, which suggests that the binding of streptomycin decreases the degree of organization of ribosomal RNA.