RESUMO
BACKGROUND: Neuroinflammation involves the activation of glial cells in neurodegenerative diseases such as Alzheimer's disease (AD). Plasmalogens (Pls) are glycerophospholipids constituting cellular membranes and play significant roles in membrane fluidity and cellular processes such as vesicular fusion and signal transduction. METHODS: In this study the preventive effects of Pls on systemic lipopolysaccharide (LPS)-induced neuroinflammation were investigated using immunohistochemistry, real-time PCR methods and analysis of brain glycerophospholipid levels in adult mice. RESULTS: Intraperitoneal (i.p.) injections of LPS (250 µg/kg) for seven days resulted in increases in the number of Iba-1-positive microglia and glial fibrillary acidic protein (GFAP)-positive astrocytes in the prefrontal cortex (PFC) and hippocampus accompanied by the enhanced expression of IL-1ß and TNF-α mRNAs. In addition, ß-amyloid (Aß3-16)-positive neurons appeared in the PFC and hippocampus of LPS-injected animals. The co-administration of Pls (i.p., 20 mg/kg) after daily LPS injections significantly attenuated both the activation of glial cells and the accumulation of Aß proteins. Finally, the amount of Pls in the PFC and hippocampus decreased following the LPS injections and this reduction was suppressed by co-treatment with Pls. CONCLUSIONS: These findings suggest that Pls have anti-neuroinflammatory and anti-amyloidogenic effects, thereby indicating the preventive or therapeutic application of Pls against AD.
Assuntos
Peptídeos beta-Amiloides/antagonistas & inibidores , Edema Encefálico/tratamento farmacológico , Lipopolissacarídeos/toxicidade , Plasmalogênios/uso terapêutico , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Galinhas , Glicerofosfolipídeos/uso terapêutico , Inflamação/tratamento farmacológico , Inflamação/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
BACKGROUND: Many disorders with plasmalogen deficiency have been reported. Replenishment or replacement of tissue plasmalogens of these disorders would be beneficial to the patients with these disorders, but effects of dietary plasmalogen on mammals have not been reported. METHODS: Plasmalogens were purified from chicken skin. The purified plasmalogens consisted of 96.4% ethanolamine plasmalogen (PlsEtn), 2.4% choline plasmalogen (PlsCho) and 0.5% sphingomyelin (SM). A diet containing 0.1% the purified plasmalogens (PlsEtn diet) was given to rats. Relative composition of phospholipids was measured by a high performance liquid chromatography (HPLC) method that can separate intact plasmalogens and all other phospholipid classes by a single chromatographic run. RESULTS: The PlsEtn diet given to Zucker diabetic fatty (ZDF) rats for 4 weeks caused decreases of plasma cholesterol and plasma phospholipid as compared to control diet. The other routine laboratory tests of plasma including triacylglycerol, glucose, liver and renal functions, albumin, and body weight were not different. Relative compositions of erythrocyte PlsEtn and phosphatidylethanolamine (PE) increased, and that of phosphatidylcholine (PC) decreased in PlsEtn diet group. The PlsEtn diet given to normal rats for 9 weeks again caused decrease of plasma cholesterol and phospholipid, and it induced increase of relative composition of PlsEtn of the erythrocyte membrane. The other routine laboratory tests of plasma and body weight were not different. CONCLUSIONS: Dietary PlsEtn increases relative composition of PlsEtn of erythrocyte membranes in normal and ZDF rats, and it causes decreases of plasma cholesterol and plasma phospholipids. Dietary PlsEtn for 9 weeks seemingly causes no adverse effect to health of normal rats.
Assuntos
Membrana Eritrocítica/metabolismo , Plasmalogênios/administração & dosagem , Plasmalogênios/sangue , Animais , Colesterol/sangue , Suplementos Nutricionais , Masculino , Fosfatidilcolinas/sangue , Fosfatidiletanolaminas/sangue , Fosfolipídeos/sangue , Ratos , Ratos ZuckerRESUMO
A standard plasmid was constructed as a novel reference molecule for use in real-time quantitative PCR assays to verify the identity of beef, pork, chicken, mutton, and horseflesh. The plasmid contained a target domain of the cytochrome b (cyt b) gene and an artificial DNA sequence. Primers CO-F and CO-R, and probe CO-P were specifically designed to detect the artificial sequence. The calculated R² values of the standard curves (10³-107 copies per reaction) for the five species ranged between 0.998 and 0.999 in the quantification analysis. The constructed plasmid provides a universal method for measuring the copy number of cyt b DNA in minced meat. This method would be a useful procedure for verifying food labels.
Assuntos
Bioensaio , Citocromos b/genética , Primers do DNA/genética , Sondas de DNA/análise , Produtos da Carne/análise , Plasmídeos/análise , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Sequência de Bases , Bovinos , Galinhas , Citocromos b/química , Primers do DNA/química , Sondas de DNA/química , Sondas de DNA/genética , Escherichia coli , Dosagem de Genes , Genes Sintéticos , Cavalos , Dados de Sequência Molecular , Plasmídeos/química , Plasmídeos/genética , Ovinos , SuínosRESUMO
To manufacture raw ham in an efficient manner, we recently developed a new system in which presliced pork loin was used, and the processing time was reduced to 5% of the conventional method. This study aimed to examine whether this raw ham could be as safe as ham produced by the conventional method. Pork loin spiked with enterohemorrhagic Escherichia coli serotype O157:H7, Listeria monocytogenes serotype 1/2c, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus were processed using either the new or conventional method. The fate of the foodborne pathogens and behavior of hygiene indicator bacteria were examined. Whereas nitrite had disappeared during the conventional packaging process, the reduced processing time in the new system allowed for the ham to be vacuum packed with retention of the nitrite (6.9±1.2 ppm, P<0.01). This accounts for the prominent decrease in L. monocytogenes (2.3 log reduction in 35 days) and S. aureus (3.3 log reduction in 13 days) counts during storage. E. coli O157 and Salmonella Enteritidis were likely resistant to the nitrite in the ham. However, they were unable to multiply in the ham and decreased gradually as in the conventionally produced ham. The bacteriostatic nature of the raw ham was also indicated by the gradual decrease in coliforms (1.3 log reduction in 13 days) in nonspiked ham. In conclusion, the raw ham produced using presliced pork loin is practically as safe as conventionally produced raw ham. It is worth validating these results in a small-scale production setting.
Assuntos
Qualidade de Produtos para o Consumidor , Manipulação de Alimentos/métodos , Carne/microbiologia , Viabilidade Microbiana , Animais , Contagem de Colônia Microbiana , Escherichia coli O157/crescimento & desenvolvimento , Microbiologia de Alimentos , Humanos , Concentração de Íons de Hidrogênio , Listeria monocytogenes/crescimento & desenvolvimento , Salmonella enteritidis/crescimento & desenvolvimento , Staphylococcus aureus/crescimento & desenvolvimento , Suínos , Fatores de TempoRESUMO
The aim of this study was to evaluate the effect of lactic acid (LA) with and without organic material at various post-treatment recovery times on the heat resistance of Listeria monocytogenes (Lm). LA decreased Lm numbers; however, the effect was remarkably attenuated by the presence of organic matter. Five strains of Lm were treated with LA and the listericidal effects were compared. The effect of LA varied depending on the strain, with ≥3.0% (w/w) LA required to kill the Lm strains in a short time. The heat resistance of Lm treated with LA was examined with respect to the time interval between the acid treatment and the subsequent manufacturing step. The heat resistance of Lm was shown to significantly increase during the post-treatment period. Heat tolerance (D value) increased up to 3.4-fold compared with the non-treated control bacteria. RNA sequencing and RT-PCR analyses suggested that several stress chaperones, proteins controlled by RecA and associated with high-temperature survival, were involved in the mechanism of enhanced heat resistance. These results are applicable to manufacturers when LA and heat treatment methods are utilized for the effective control of Lm in foods.
Assuntos
Ácido Láctico/farmacologia , Listeria monocytogenes/efeitos dos fármacos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Contagem de Colônia Microbiana , Temperatura Alta , Listeria monocytogenes/química , Listeria monocytogenes/crescimento & desenvolvimento , Listeria monocytogenes/isolamento & purificação , Fatores de TempoRESUMO
Neuronal cells are susceptible to many stresses, which will cause the apoptosis and neurodegenerative diseases. The precise molecular mechanism behind the neuronal protection against these apoptotic stimuli is necessary for drug discovery. In the present study, we have found that plasmalogens (Pls), which are glycerophospholipids containing vinyl ether linkage at sn-1 position, can protect the neuronal cell death upon serum deprivation. Interestingly, caspse-9, but not caspase-8 and caspase-12, was cleaved upon the serum starvation in Neuro-2A cells. Pls treatments effectively reduced the activation of caspase-9. Furthermore, cellular signaling experiments showed that Pls enhanced phosphorylation of the phosphoinositide 3-kinase (PI3K)-dependent serine/threonine-specific protein kinase AKT and extracellular-signal-regulated kinases ERK1/2. PI3K/AKT inhibitor LY294002 and MAPK/ERK kinase (MEK) inhibitor U0126 treatments study clearly indicated that Pls-mediated cell survival was dependent on the activation of these kinases. In addition, Pls also inhibited primary mouse hippocampal neuronal cell death induced by nutrient deprivation, which was associated with the inhibition of caspase-9 and caspase-3 cleavages. It was reported that Pls content decreased in the brain of the Alzheimer's patients, which indicated that the reduction of Pls content could endanger neurons. The present findings, taken together, suggest that Pls have an anti-apoptotic action in the brain. Further studies on precise mechanisms of Pls-mediated protection against cell death may lead us to establish a novel therapeutic approach to cure neurodegenerative disorders.
Assuntos
MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Neurônios/citologia , Neurônios/efeitos dos fármacos , Plasmalogênios/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Caspases/metabolismo , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Hipocampo/citologia , Camundongos , Neurônios/metabolismo , Fosforilação/efeitos dos fármacos , Proteólise/efeitos dos fármacos , Tretinoína/farmacologiaRESUMO
Neuroinflammation essentially involves an activation of glial cells as the cause/effect of neurodegenerative diseases such as Alzheimer's disease (AD). Plasmalogens (Pls) are glycerophospholipids constituting cellular membranes and play significant roles in membrane fluidity and cellular processes like vesicular fusion and signal transduction. Intraperitoneal (i.p.) injection of lipopolysaccharide (LPS, 250 µg/kg) for 7 days resulted in the morphological changes and increase in number of Iba-1(+) microglia showing neuroinflammation in the adult mouse hippocampus. The LPS-induced activation of glial cells was significantly attenuated by i.p. pretreatment with Pls dissolved in corn oil. In addition, systemic injection of LPS induced Aß(1-16) (+) neurons in the hippocampus were also abolished by application of Pls. Finally, contents of Pls in the hippocampus decreased after LPS injection, and the reduction was suppressed by administration of Pls. These findings suggest an antiamyloidogenic effect of Pls, implicating a possible therapeutic application of Pls against AD.