RESUMEN
The periparturient period is a metabolically demanding time for dairy animals because of increased nutrient requirements for milk yield. The objective of this study was to investigate the effect of feeding Saccharomyces cerevisiae boulardii (CNCM I-1079), a commercial active dry yeast (ADY), in dairy cows on productive and metabolic measures during the periparturient period. Primiparous (n = 33) and multiparous (n = 35) cows were fed a close-up total mixed ration (TMR) before calving and a lactation TMR postpartum. Three weeks before expected calving time, animals were blocked by parity and body weight and then randomly assigned to either control group (control; n = 34) or treatment (ADY; n = 34). All animals were housed in a tie-stall barn with individual feed bunks; the ADY animals received supplementary Saccharomyces cerevisiae boulardii (CNCM I-1079), top dressed daily at a predicted dosage of 1.0 × 1010 cfu (12.5 g) per head. Blood samples were collected weekly along with milk yield and milk composition data; feed intake data were collected daily. Serum samples were analyzed for glucose, nonesterified fatty acid, ß-hydroxybutyrate, haptoglobin (Hp), and the cytokines tumor necrosis factor-α, IL-6, and IL-18. Colostrum samples collected within the first 6 to 10 h were analyzed for somatic cell score and IgG, IgA, and IgM concentrations. Data were analyzed using PROC GLIMMIX in SAS with time as a repeated measure; model included time, parity, treatment, and their interactions. The ADY groups had greater milk yield (39.0 ± 2.4 vs. 36.7 ± 2.3 kg/d) and tended to produce more energy-corrected milk with better feed efficiency. There was no difference in plasma glucose, serum nonesterified fatty acid, serum ß-hydroxybutyrate, Hp, IL-6, or IL-18 due to ADY treatment. The tumor necrosis factor-α increased in ADY-supplemented animals (1.17 ± 0.69 vs. 4.96 ± 7.7 ng/mL), though week, parity, and their interactions had no effect. Serum amyloid A tended to increase in ADY-supplemented animals when compared to control animals and was additionally affected by week and parity; there were no significant interactions. No difference in colostrum IgG, IgA, and IgM was observed between treatments. Supplementing transition cow TMR with ADY (CNCM I-1079) improved milk production and tended to improve efficiency in early lactation; markers of inflammation were also influenced by ADY treatment, though the immunological effect was inconsistent.
Asunto(s)
Saccharomyces boulardii , Saccharomyces cerevisiae , Embarazo , Femenino , Bovinos , Animales , Saccharomyces cerevisiae/metabolismo , Interleucina-18/metabolismo , Ácido 3-Hidroxibutírico , Factor de Necrosis Tumoral alfa/metabolismo , Interleucina-6/metabolismo , Dieta/veterinaria , Metabolismo Energético , Lactancia , Leche/metabolismo , Ingestión de Alimentos , Periodo Posparto/metabolismo , Ácidos Grasos no Esterificados , Inmunoglobulina A/metabolismo , Inmunoglobulina G/metabolismo , Inmunoglobulina M , Alimentación Animal/análisisRESUMEN
BACKGROUND AND OBJECTIVE: The present study investigated the association between the RAGE G82S polymorphism, the plasma levels of sRAGE and chronic periodontitis in subjects with and without diabetes mellitus (DM). MATERIAL AND METHODS: A total of 230 patients with DM and 264 non-DM participants were recruited for this study. Genotyping of the RAGE G82S polymorphism was accomplished using polymerase chain reaction-restriction fragment length polymorphism, and associations were analyzed with the chi-squared test and logistic regression analysis. RESULTS: In the non-DM group, the chi-squared test showed that the frequency distributions of the G82S polymorphism were significantly different between chronic periodontitis and non-chronic periodontitis subjects (χ(2) = 8.39, p = 0.02). A multivariate logistic regression model showed that the (G82S + S82S) genotypes were associated with a significantly increased risk of chronic periodontitis development compared to the G82G genotype (adjusted odds ratio = 2.06, 95% confidence interval: 1.08-4.07). In the DM group, there was no association between the G82S polymorphism and chronic periodontitis development when a multivariate logistic regression was performed. Plasma levels of sRAGE were significantly higher in subjects with the G82G genotype compared to those with the (G82S + S82S) genotypes in both the non-DM (856.6 ± 332.0 vs. 720.4 ± 311.4 pg/mL, p = 0.003) and DM groups (915.3 ± 497.1 vs. 603.5 ± 298.3 pg/mL, p < 0.0001). However, there was no difference in plasma sRAGE levels between chronic periodontitis and non-chronic periodontitis subjects in both the DM and non-DM groups. Moreover, when the subjects were further sub-divided by the G82S polymorphism, the difference in plasma levels of sRAGE between chronic periodontitis and non-chronic periodontitis subjects in the DM and non-DM groups remained statistically insignificant. CONCLUSIONS: The present study revealed that the RAGE G82S polymorphism was associated with chronic periodontitis in the non-DM group but not in the DM group. Our results also showed that the plasma levels of sRAGE were significantly higher in subjects with the RAGE G82G genotype, and this correlation was not affected by the presence of chronic periodontitis in the DM and non-DM groups.
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Antígenos de Neoplasias/genética , Periodontitis Crónica/epidemiología , Periodontitis Crónica/genética , Complicaciones de la Diabetes , Predisposición Genética a la Enfermedad , Proteínas Quinasas Activadas por Mitógenos/genética , Polimorfismo Genético , Adulto , Sustitución de Aminoácidos , Antígenos de Neoplasias/sangre , Femenino , Técnicas de Genotipaje , Humanos , Masculino , Persona de Mediana Edad , Proteínas Quinasas Activadas por Mitógenos/sangre , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Taiwán/epidemiologíaRESUMEN
OBJECTIVES: We investigated the association between mRNA levels, polymorphisms of Kallikrein7 (KLK7) and Kallikrein10 (KLK10), and the development of oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: We recruited 217 OSCC patients and 138 healthy controls. All were men, betel quid chewers, cigarette smokers, and Minnan ethnicity. Genotyping was performed using a TaqMan probe genotyping assay. Gene expression levels were determined using real-time polymerase chain reactions (PCRs) for 20 pairs of cancerous and non-cancerous tissues. RESULTS: Kallikrein10 rs3745535G>T polymorphisms were significantly associated with OSCC development [adjusted OR (AOR) = 1.62, 95% CI = 1.02-2.59], but KLK7 polymorphisms were not. The KLK7 rs10581213(wt/ins + ins/ins) genotypes were significantly associated with early-stage cancer (AOR = 0.34, 95% CI = 0.14-0.78), but KLK10 polymorphisms were not. Relative expression analysis indicated that an increase in KLK7 and KLK10 mRNA levels was found in cancerous tissues (2(-ΔΔCT) = 25.23 ± 8.85 and 10.89 ± 4.97, respectively). A significantly higher level of KLK7 was expressed in early-stage cancer with the rs10581213(wt/ins + ins/ins) genotypes, but there was no significant difference in the mRNA levels of KLK7 and KLK10 between early- and advanced-stage cancers. CONCLUSIONS: This is the first correlation of OSCC with KLK10 rs3745535G>T polymorphisms. Early-stage OSCC and high KLK7 mRNA levels were correlated with the rs10581213(wt/ins + ins/ins) genotypes. More studies with large sample sizes are needed to verify our findings.
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Areca , Carcinoma de Células Escamosas/genética , Calicreínas/genética , Neoplasias de la Boca/genética , Polimorfismo Genético , Fumar , Areca/efectos adversos , Humanos , Masculino , Persona de Mediana Edad , Medición de Riesgo , Fumar/efectos adversos , TaiwánRESUMEN
BACKGROUND AND OBJECTIVE: The association between psychosocial factors and periodontal disease has been widely reported and might be modified by smoking status. This study investigated the association of periodontal status with psychosocial factors and smoking in a community population. MATERIAL AND METHODS: A structured questionnaire was administered to a total of 1,764 civilian noninstitutional (general population excluding from nursing homes, sanitariums and hospitals) Taiwanese individuals to assess the presence and severity of psychosocial factors [using the 12-item Chinese health questionnaire (CHQ-12)], smoking habits and other related factors. Periodontal status was established using the community periodontal index and by measuring clinical loss of attachment. RESULTS: Psychological factors and smoking were significantly associated with loss of attachment (odds ratio = 1.69, 95% confidence interval = 1.01-2.77, comparing the CHQ-12 score of >or= 6 with the CHQ-12 score of 0-2 and p = 0.032 for linear trend; odds ratio = 2.21, 95% confidence interval = 1.45-3.37, comparing smokers with nonsmokers) but not with community periodontal index. The association was found to be stronger among smokers than among nonsmokers. Smokers with a CHQ-12 score of >or= 6 had a higher odds ratio of loss of attachment (odds ratio = 2.49, 95% confidence interval = 0.91-6.49) than nonsmokers (odds ratio = 1.43, 95% confidence interval = 0.76-2.58). For periodontal health measured using the community periodontal index, married and divorced/widowed subjects tended to have poorer periodontal health (odds ratio = 3.38, 95% confidence interval = 1.26-10.81 and odds ratio = 3.83, 95% confidence interval = 1.21-13.83, respectively) than single subjects among nonsmokers but not among smokers. CONCLUSION: Poor mental health had a stronger association with periodontal disease among smokers than among nonsmokers, especially in accumulative attachment loss. Our findings suggest that mental health and smoking might have a synergistic effect on the risk of developing periodontal disease.
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Enfermedades Periodontales/etiología , Índice Periodontal , Fumar/fisiopatología , Estrés Psicológico/complicaciones , Adolescente , Adulto , Factores de Edad , Anciano , Actitud Frente a la Salud , Susceptibilidad a Enfermedades , Escolaridad , Femenino , Hemorragia Gingival/clasificación , Conductas Relacionadas con la Salud , Humanos , Masculino , Estado Civil , Salud Mental , Persona de Mediana Edad , Pérdida de la Inserción Periodontal/clasificación , Enfermedades Periodontales/clasificación , Bolsa Periodontal/clasificación , Factores de Riesgo , Factores Sexuales , Encuestas y Cuestionarios , Pérdida de Diente/clasificación , Adulto JovenRESUMEN
ABSTRACT Infective endocarditis is less likely to sparkle out preferentially in our minds when evaluating and making differential diagnosis of patients with fever daily in emergency departments. We describe a case of infective endocarditis. He was initially diagnosed with pyelonephritis of the right kidney at a hospital because of the noted right flank knocking pain. His computed tomography showed two wedge-shaped low-density lesions in the spleen and the right kidney separately. It dropped a hint to the emergency department physician of thinking of the feature of infarct. The previously neglected cardiac murmurs were then an important clue. We then performed transthoracic emergent echocardiography and confirmed the diagnosis of infective endocarditis.
RESUMEN
The binding mechanism of Mg(2+) at the M3 site of human placental alkaline phosphatase was found to be a slow-binding process with a low binding affinity (K(Mg(app.)) = 3.32 mM). Quenching of the intrinsic fluorescence of the Mg(2+)-free and Mg(2+)-containing enzymes by acrylamide showed almost identical dynamic quenching constant (K(sv) = 4.44 +/- 0.09 M(-1)), indicating that there is no gross conformational difference between the M3-free and the M3-Mg(2+) enzymes. However, Zn(2+) was found to have a high affinity with the M3 site (K(Zn(app.)) = 0.11 mM) and was observed as a time-dependent inhibitor of the enzyme. The dependence of the observed transition rate from higher activity to lower activity (k(obs)) at different zinc concentrations resulted in a hyperbolic curve suggesting that zinc ion induces a slow conformational change of the enzyme, which locks the enzyme in a conformation (M3'-Zn) having an extremely high affinity for the Zn(2+) (K*(Zn(app.)) = 0.33 microM). The conformation of the M3'-Zn enzyme, however, is unfavorable for the catalysis by the enzyme. Both Mg(2+) activation and Zn(2+) inhibition of the enzyme are reversible processes. Structural information indicates that the M3 site, which is octahedrally coordinated to Mg(2+), has been converted to a distorted tetrahedral coordination when zinc ion substitutes for magnesium ion at the M3 site. This conformation of the enzyme has a small dynamic quenching constant for acrylamide (K(sv) = 3.86 +/- 0.04 M(-1)), suggesting a conformational change. Both Mg(2+) and phosphate prevent the enzyme from reaching this inactive structure. GTP plays an important role in reactivating the Zn-inhibited enzyme activity. We propose that, under physiological conditions, magnesium ion may play an important modulatory role in the cell for protecting the enzyme by retaining a favorable geometry of the active site needed for catalysis.
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Fosfatasa Alcalina/metabolismo , Magnesio/farmacología , Placenta/enzimología , Zinc/farmacología , Fosfatasa Alcalina/química , Fosfatasa Alcalina/efectos de los fármacos , Sitios de Unión , Interacciones Farmacológicas , Activación Enzimática/efectos de los fármacos , Estabilidad de Enzimas , Humanos , Fosfatos/farmacología , Sustancias Protectoras/farmacología , Conformación Proteica , Factores de TiempoRESUMEN
1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is known to produce a differential toxicity in the nigrostriatal and mesolimbic dopaminergic pathways with the nigrostriatal pathway being more vulnerable. We, therefore, investigated whether oxidative stress and the antioxidant system play a role in this phenomenon. Balb/c mice were treated with either saline or MPTP (30 mg/kg/d) for 7 d, and were sacrificed on the next day. Results revealed that MPTP increased lipid peroxidation in the striatum (ST) and decreased glutathione concentration in the substantia nigra (SN) without markedly affecting these measures in the nucleus accumbens (NAc) and ventral tegmental area (VTA). Further, MPTP produced approximately twofold increases in both manganese superoxide dismutase (MnSOD) and copper-zinc superoxide dismutase (CuZnSOD) activities in the VTA while it only increased MnSOD activity in the SN. Both catalase and glutathione peroxidase (GPx) activities were not markedly altered by MPTP in both systems. However, the basal levels of catalase and GPx activities were higher in the VTA and NAc than in the SN and ST. These results together suggest that a lesser degree of oxidative damage and a more inducible CuZnSOD activity observed in the mesolimbic dopaminergic pathway may partially explain the differential toxicity MPTP produced in these two dopaminergic systems.
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Antioxidantes/metabolismo , Encéfalo/efectos de los fármacos , Dopaminérgicos/toxicidad , Dopamina/fisiología , Intoxicación por MPTP , Estrés Oxidativo , Animales , Encéfalo/metabolismo , Catalasa/metabolismo , Cuerpo Estriado/efectos de los fármacos , Glutatión/metabolismo , Glutatión Peroxidasa/metabolismo , Sistema Límbico/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos BALB C , Sustancia Negra/efectos de los fármacos , Superóxido Dismutasa/metabolismoRESUMEN
Both genetic and environmental factors are involved in the development of cancer; some phase I and II enzymes involved in the metabolism of carcinogens are polymorphic in genotypes. This case-control study focused on the interactions between oral cancer risk factors and genetic polymorphisms of cytochrome P-450 (CYP) 2E1 and glutathione S-transferase (GST) M1 and GSTT1. A total of 41 male oral cancer cases was recruited from National Taiwan University Hospital, and 123 healthy controls frequency-matched on ethnicity, sex, and age were recruited from residents living in Taipei City and Taipei County. History of cigarette smoking, alcohol drinking, and betel quid chewing was obtained through a standardized questionnaire interview, and genotypes of CYP2E1, GSTM1, and GSTT1 were determined by PCR. Cigarette smoking, alcohol drinking, and betel quid chewing were significantly associated with the risk of oral cancer in a dose-response relationship. All betel quid chewers smoked cigarettes in both the case and control groups. In the multiple logistic regression analysis, those who had null genotypes of GSTM1 and/or GSTT1 had an increased oral cancer risk compared with those who had non-null genotypes of both GSTM1 and GSTT1, showing a multivariate-adjusted odds ratio (OR) of 4.6 with a 95% confidence interval (CI) of 0.9-23.7 (P = 0.08). The CYP2E1 c1/c2 and c2/c2 genotypes were associated with a significantly increased oral cancer risk compared with the c1/c1 genotype among those who did not chew betel quid (OR, 4.7; 95% CI, 1.1-20.2), but not among betel quid chewers. Habitual alcohol drinking was associated with a significantly increased oral cancer risk, showing an OR of 3.0 (95% CI, 1.1-8.8). These results implied that there are gene-gene and gene-environment interactions in the development of oral cancer.
Asunto(s)
Citocromo P-450 CYP2E1/genética , Glutatión Transferasa/genética , Neoplasias de la Boca/genética , Consumo de Bebidas Alcohólicas , Areca , Estudios de Casos y Controles , Exposición a Riesgos Ambientales , Genotipo , Humanos , Modelos Logísticos , Masculino , Neoplasias de la Boca/enzimología , Neoplasias de la Boca/epidemiología , Análisis Multivariante , Plantas Medicinales , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Factores de Riesgo , Fumar , Taiwán/epidemiologíaRESUMEN
In the present study we examined the role of BDNF gene expression involved in the differential vulnerability of the nigrostriatal and mesolimbic dopaminergic pathways to environmental damage. The toxins for dopamine (DA) neurons 1-methyl-4-phenyl-1,2,3,6,-tetrahydropyridine (MPTP) and 1-methyl-4-phenylpyridinium (MPP+) were used as pharmacological tools. Results revealed that chronic MPTP treatment produced a significant and irreversible DA depletion in the striatum (ST) as well as a marked decrease in tyrosine-hydroxylase (TH) mRNA level in the substantia nigra (SN). Under these conditions, the endogenous brain-derived neurotrophic factor (BDNF) mRNA level was increased in the SN. Only acute DA reduction was found in the nucleus accumbens (NAc) and TH mRNA level was not affected in the ventral tegmental area (VTA) by MPTP treatment. Further, when MPP+ produced a similar extent of DA depletion in the ST and NAc, the TH mRNA level was also decreased while BDNF mRNA level was increased in the SN. The same alterations were not observed in the VTA. Results from the BDNF mRNA regional distribution study revealed that structures in the mesolimbic dopaminergic pathway expressed a more than 2-fold higher basal BDNF mRNA level than structures in the nigrostriatal dopaminergic pathway. Presumably, enhanced BDNF gene expression would help the survival of DA neurons and these findings suggest a better protective mechanism in the mesolimbic pathway. Lastly, direct BDNF infusions to the SN partially protected against MPTP's toxicity on DA neurons in the ST in mice. These results together suggest that a more abundant BDNF mRNA level along the mesolimbic pathway than the nigrostriatal pathway may, at least partially, explain the differential vulnerability of different DA neurons to MPTP and MPP+ toxicity.
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1-Metil-4-fenilpiridinio/toxicidad , Factor Neurotrófico Derivado del Encéfalo/fisiología , Cuerpo Estriado/efectos de los fármacos , Dopaminérgicos/toxicidad , Dopamina/fisiología , Sistema Límbico/efectos de los fármacos , Intoxicación por MPTP , Proteínas del Tejido Nervioso/biosíntesis , Vías Nerviosas/efectos de los fármacos , Sustancia Negra/efectos de los fármacos , Tirosina 3-Monooxigenasa/biosíntesis , Animales , Factor Neurotrófico Derivado del Encéfalo/biosíntesis , Factor Neurotrófico Derivado del Encéfalo/genética , Cuerpo Estriado/fisiología , Dopamina/biosíntesis , Inducción Enzimática/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Sistema Límbico/fisiología , Masculino , Ratones , Ratones Endogámicos BALB C , Proteínas del Tejido Nervioso/genética , Núcleo Accumbens/metabolismo , Especificidad de Órganos , Reacción en Cadena de la Polimerasa , ARN Mensajero/biosíntesis , Sustancia Negra/fisiología , Tirosina 3-Monooxigenasa/genéticaRESUMEN
This study examines the roles of PKC and protein phosphorylation in the retention performance of a passive avoidance learning (PAL) task in rats. Results revealed that H7 injected into the dentate gyrus (DG) of the hippocampus impaired retention in a dose-dependent manner. Corticotropin-releasing factor (CRF) injected into the DG improved retention and this facilitation was antagonized by H7 pretreatment. CRF increased phosphorylation of five proteins, whereas H7 decreased phosphorylation in three of these proteins in both the cytosol and the membrane fractions of hippocampus. These effects were shown not to be associated with stress. These results demonstrate that CRF increased protein phosphorylation associated with enhanced retention of PAL task in rats.
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Reacción de Prevención/fisiología , Hormona Liberadora de Corticotropina/fisiología , Hipocampo/fisiología , Proteínas del Tejido Nervioso/metabolismo , Proteína Quinasa C/fisiología , Retención en Psicología/fisiología , 1-(5-Isoquinolinesulfonil)-2-Metilpiperazina , Animales , Isoquinolinas/farmacología , Masculino , Fosforilación , Piperazinas/farmacología , Proteína Quinasa C/antagonistas & inhibidores , Ratas , Ratas EndogámicasRESUMEN
The present study used pharmacological, biochemical, and behavioral methods to examine the role of protein synthesis in the hippocampus in memory processes of a passive avoidance learning in rats. Results indicated that corticotropin-releasing factor (CRF) significantly improved memory retention in rats. Both cycloheximide (CHX) and actinomycin-D (ACT-D) impaired memory at high doses. At doses of CHX and ACT-D that did not affect memory alone, they both antagonized the memory-enhancing effect of CRF. Biochemically, there were specific increases in the optical density of three protein bands in the cytosolic fraction of hippocampal cells in rats showing good memory. There were also marked increases in the optical density of two protein bands in the nucleus fraction of the same animals. Similar results were observed in animals injected with CRF. However, no significant protein alteration was observed in animals receiving stress. These results together suggest that there are new protein syntheses in the hippocampus that are specifically associated with passive avoidance learning in rats.
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Hormona Liberadora de Corticotropina/farmacología , Hipocampo/efectos de los fármacos , Memoria/efectos de los fármacos , Proteínas del Tejido Nervioso/biosíntesis , Animales , Reacción de Prevención/efectos de los fármacos , Reacción de Prevención/fisiología , Hormona Liberadora de Corticotropina/administración & dosificación , Cicloheximida/farmacología , Dactinomicina/farmacología , Hipocampo/anatomía & histología , Hipocampo/metabolismo , Masculino , Memoria/fisiología , Proteínas del Tejido Nervioso/aislamiento & purificación , Ratas , Ratas Sprague-DawleyRESUMEN
The present study examined the possible differential toxicity of 1-methyl-4-phenyl-pyridinium (MPP+) in the nigrostriatal and mesolimbic dopaminergic pathways in BALB/c mice, and investigated whether MPP+ uptake may account for this differential toxicity. Results indicated that chronic MPP+ infusions to the striatum (ST) and nucleus accumbens (NAc) both produced a toxicity on dopamine (DA) neurons. However, MPP+ produced a more severe DA depletion in the ST than in the NAc. Kinetic analyses from MPP+ uptake studies revealed a similar Km value in both the ST and NAc, suggesting that the affinity for MPP+ uptake is not different. The Vmax was approximately 1.6-fold higher in the ST than in the NAc. These results together suggest that chronic MPP+ infusions produce a differential toxicity on DA neurons in the nigrostriatal and mesolimbic dopaminergic pathways and that the ability of MPP+ uptake in DA terminals probably does not account for this differential toxicity.
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1-Metil-4-fenilpiridinio/metabolismo , 1-Metil-4-fenilpiridinio/farmacología , Cuerpo Estriado/efectos de los fármacos , Dopamina/metabolismo , Núcleo Accumbens/efectos de los fármacos , Animales , Relación Dosis-Respuesta a Droga , Cinética , Masculino , Ratones , Ratones Endogámicos BALB C , Vías Nerviosas/efectos de los fármacosRESUMEN
Studies of the dietary phytoestrogen biochanin A on cell proliferation of the cultured estrogen responsive cells human breast carcinoma MCF-7 showed that biochanin A exhibits biphasic regulation on MCF-7 cells. At concentrations of less than 10 microg/mL, cells respond to biochanin A by increasing cell growth and de novo DNA synthesis. The addition of biochanin A at concentrations of greater than 30 microg/mL significantly inhibited cell growth and DNA synthesis in a dose-dependent fashion, resulting in an IC(50) value of 40 microg/mL. The reversibility of these inhibitory effects by biochanin A appears also to be concentration dependent. Cells previously treated with high concentrations (>60 microg/mL) of biochanin A did not regain normal growth after treatment ceased. Biochanin A was cytostatic at low concentrations (<40 microg/mL) and cytotoxic at higher concentrations. Upon exposure to 100 microg/mL of biochanin A, cell morphology was severely altered, cell volume decreased, and condensation of cell components was clearly noticeable. In addition, biochanin A damaged cell membranes by increasing membrane permeability. These results suggest possible molecular and cellular mechanisms of the action of dietary phytoestrogens on estrogen target cells.
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The hematopoietic growth factor, granulocyte colony-stimulating factor (G-CSF), has become one of the few growth factors approved for clinical use. It has therapeutic potential for numerous neurodegenerative diseases; however, at present the cellular effects of G-CSF on the central nervous system remain unclear and in need of investigation. In the present study, we used spinal cord ischemia, a neurodegenerative model, to examine the effects of intrathecal (i.t.) G-CSF on glial cell (microglia and astrocyte) activation and neuroprotective factor expression, including glial cell line-derived neurotrophic factor (GDNF) and vascular endothelial growth factor A (VEGF-A) protein expression. Our results indicate that i.t. G-CSF could enhance ischemia-induced microglial activation and inhibit ischemia-induced astrocyte activation. Both GDNF and VEGF-A are upregulated after injury, and i.t. G-CSF could enhance GDNF and VEGF-A expressions after injury. Interestingly, our results indicate that performing i.t. G-CSF alone on normal animals could have the effect of microglial and astrocyte activation and enhanced GDNF and VEGF-A expressions. Furthermore, through laser scanning confocal microscopy, we found that astrocytes may contribute to the majority of GDNF and VEGF-A expressions of G-CSF after spinal cord ischemia. Overall, this G-CSF-induced upregulation suggests that activation of endogenous neuroprotective mechanisms could resist neurodegenerative insults. These observations demonstrate the cellular mechanism of i.t. G-CSF after spinal cord ischemia and confirm the neuroprotective effect of G-CSF after spinal cord ischemia injury.
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Astrocitos/efectos de los fármacos , Factor Neurotrófico Derivado de la Línea Celular Glial/biosíntesis , Factor Estimulante de Colonias de Granulocitos/administración & dosificación , Factor Estimulante de Colonias de Granulocitos/farmacología , Isquemia/metabolismo , Médula Espinal/patología , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Animales , Astrocitos/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Factor Estimulante de Colonias de Granulocitos/uso terapéutico , Inyecciones Espinales , Isquemia/tratamiento farmacológico , Isquemia/patología , Masculino , Microglía/efectos de los fármacos , Fármacos Neuroprotectores/administración & dosificación , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Ratas , Ratas Wistar , Recuperación de la Función , Médula Espinal/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Nonketotic hyperglycemia (NKH) is a clinical syndrome consisting of hyperglycemia, hyperosmolality and intracellular dehydration but not ketoacidosis. This prospective study evaluated the clinical and magnetic resonance imaging abnormalities in six patients with NKH complicated with simple or complex partial seizures. Subcortical T2 hypointensity rather than hyperintensity together with contrast enhancement was a characteristic feature of seizures associated with NKH. Restricted diffusion on DWI and decreased NAA and/or Choline on MRS studies were also noted.
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The aim of this study was to examine the relationship between placental polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDD/F) and polychlorinated biphenyls (PCBs) toxic equivalent (TEQ) levels and the consumption of various food types in pregnant women from central Taiwan. Placental PCDD/Fs and PCB congener TEQ levels were evaluated in 109 pregnant women and dietary information was obtained by questionnaire. TEQ levels of PCDD/Fs and PCBs were positively associated with age and annual family incomes (p < 0.05). PCDD/F TEQs were significantly associated with freshwater fish and dairy product consumption after adjustment for age and body mass index (BMI) (p < 0.05). For PCB TEQs, significant associations were detected for saltwater fish consumption (p < 0.05). In summary, positive correlations were found between freshwater fish and dairy product intake and PCDD/F levels, and a marginal correlation between saltwater fish intake and the body burden of PCBs in pregnant women from central Taiwan. Risk assessment of PCDD/Fs and PCB in fishery products is warranted in a future study to quantify the benefits of fish consumption during the perinatal period.
Asunto(s)
Benzofuranos/análisis , Dioxinas/análisis , Placenta/química , Bifenilos Policlorados/análisis , Adulto , Dieta , Exposición a Riesgos Ambientales , Contaminantes Ambientales/análisis , Femenino , Contaminación de Alimentos , Humanos , Embarazo , TaiwánRESUMEN
Peptidylarginine deiminases (PADIs) convert peptidylarginine into citrulline via posttranslational modification. One member of the family, PADI4, plays an important role in immune cell differentiation and cell death. To elucidate the participation of PADI4 in haematopoietic cell death, we examine whether inducible overexpression of PADI4 enhances the apoptotic cell death. PADI4 reduced the viability in a dose- and time-dependent manner of human leukemia HL-60 cells and human acute T leukemia Jurkat cells. The apoptosis-inducing activities were determined by nuclear condensation, DNA fragmentation, sub-G1 appearance, loss of mitochondrial membrane potential (delta psi(m)), release of mitochondrial cytochrome c into cytoplasm and proteolytic activation of caspase 9 and 3. Following PADI4 overexpression, cells arrest in G1 phase significantly before their entrance into apoptotic cell death. PADI4 increases tumor suppressor p53 and its downstream p21 to control cell cycle. In the detections of protein expression and kinase activity, all protein levels of cyclin-dependent kinases (CDKs) and cyclins are not reduced except cyclin D, however, CDK2 (G1 entry S phase) and CDK1 (G2 entry M phase) enzyme activities are inhibited by conditionally inducible PADI4. p53 also expands its other downstream Bax to induce cytochrome c release from mitochondria. According to these data, we suggest that PADI4 induces apoptosis mainly through cell cycle arrest and mitochondria-mediated pathway. Furthermore, p53 features in PADI4-induced apoptosis by increasing intracellular p21 to control cell cycle and by Bax accumulation to decline Bcl-2 function, destroy delta psi(m), release cytochrome c to cytoplasm and activate the caspase cascade.
Asunto(s)
Apoptosis/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica , Hidrolasas/metabolismo , Linfocitos T/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Células HL-60 , Humanos , Hidrolasas/farmacología , Células Jurkat , Potenciales de la Membrana/efectos de los fármacos , Mitocondrias/efectos de los fármacos , Arginina Deiminasa Proteína-Tipo 4 , Desiminasas de la Arginina ProteicaRESUMEN
Prolactin has more than 300 separate functions including affecting mammary growth, differentiation, secretion and anti-apoptosis. In the previous studies, prolactin induced Bcl-2 expression to prevent apoptosis and also provoked the activity of ornithine decarboxylase (ODC). Our previous data showed that ODC overexpression upregulates Bcl-2 and prevents tumor necrosis factor alpha (TNF-alpha)- and methotrexate (MTX)-induced apoptosis. Here, we further investigate whether prolactin prevents MTX-induced apoptosis through inducing ODC activity and the relationship between ODC and Bcl-2 upon prolactin stimulation. Prolactin prevented MTX-induced apoptosis in a dose-dependent manner in HL-60 cells. Following prolactin stimulation, ODC enzyme activity also shows an increase in a dose-dependent manner, expressing its maximum level at 3 h, and rapidly declining thereafter. Prolactin-induced ODC activity is completely blocked by a protein kinase C delta (PKCdelta) inhibitor, rottlerin. However, there are no changes in the expressions of ODC mRNA and protein level after prolactin stimulus. It indicates that prolactin may induce ODC activity through the PCKdelta pathway. Besides, Bcl-2 expresses within 1 h of prolactin treatment and this initiating effect of prolactin is not inhibited by alpha-difluoromethylornithine (DFMO). However, Bcl-2 is further enhanced following prolactin stimulation for 4 h and this enhancement is blocked by DFMO. Bcl-2 has no effect on ODC activity and protein levels, but ODC upregulates Bcl-2, which is inhibited by DFMO. Overall, there are two different forms of prolactin effect, it induces Bcl-2 primarily, and following this it stimulates ODC activity. Consequently induced ODC activity further enhances the expression of Bcl-2. The anti-apoptotic effect of prolactin is diminished by DFMO and recovered by putrescine. Obviously, ODC activity is one basis for the anti-apoptotic mechanisms of prolactin. A Bcl-2 inhibitor, HA14-1, together with DFMO, completely blocks the anti-apoptotic effects of prolactin. These results suggest that increasing ODC activity is another way of prolactin preventing MTX-induced apoptosis and that this induction of ODC activity enhances the expression of Bcl-2 strongly enough to bring about the anti-apoptotic function.
Asunto(s)
Apoptosis/fisiología , Antagonistas del Ácido Fólico/metabolismo , Metotrexato/metabolismo , Ornitina Descarboxilasa/metabolismo , Prolactina/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Apoptosis/efectos de los fármacos , Fragmentación del ADN , Relación Dosis-Respuesta a Droga , Células HL-60 , Humanos , Metotrexato/farmacología , Ornitina Descarboxilasa/genética , Prolactina/farmacología , Regulación hacia Arriba , Proteína bcl-X/metabolismoRESUMEN
Antizymes delicately regulate ornithine decarboxylase (ODC) enzyme activity and polyamine transportation. One member of the family, antizyme-1, plays vital roles in molecular and cellular functions, including developmental regulation, cell cycle, proliferation, cell death, differentiation and tumorigenesis. However, the question of how does it participate in the cell apoptotic mechanism is still unsolved. To elucidate the contribution of human antizyme-1 in haematopoietic cell death, we examine whether inducible overexpression of antizyme enhances apoptotic cell death. Antizyme reduced the viability in a dose- and time-dependent manner of human leukemia HL-60 cells, acute T leukemia Jurkat cells and mouse macrophage RAW 264.7 cells. The apoptosis-inducing activities were determined by nuclear condensation, DNA fragmentation, sub-G(1) appearance, loss of mitochondrial membrane potential (Deltapsi( m )), release of mitochondrial cytochrome c into cytoplasm and proteolytic activation of caspase 9 and 3. Following conditional antizyme overexpression, all protein levels of cyclin-dependent kinases (Cdks) and cyclins are not significantly reduced, except cyclin D, before their entrance into apoptotic cell death. However, introduced cyclin D1 into Jurkat T tetracycline (Tet)-On cell system still couldn't rescue cells from apoptosis. Antizyme doesn't influence the expression of tumor suppressor p53 and its downstream p21, but it interferes in the expressions of Bcl-2 family. Inducible antizyme largely enters mitochondria resulting in cytochrome c release from mitochondria to cytosol following Bcl-xL decrease and Bax increase. According to these data, we suggest that antizyme induces apoptosis mainly through mitochondria-mediated and cell cycle-independent pathway. Furthermore, antizyme induces apoptosis not only by Bax accumulation reducing the function of the Bcl-2 family, destroying the Deltapsi( m ), and releasing cytochrome c to cytoplasm but also by the activation of apoptosomal caspase cascade.
Asunto(s)
Apoptosis/fisiología , Caspasas/fisiología , Sistema Hematopoyético/fisiología , Potenciales de la Membrana/fisiología , Membranas Mitocondriales/fisiología , Proteínas/fisiología , Animales , Caspasa 3/metabolismo , Ciclina D , Ciclina D1/genética , Ciclina D1/metabolismo , Ciclinas/metabolismo , Citocromos c/metabolismo , Células HL-60 , Humanos , Células Jurkat , Ratones , Mitocondrias/metabolismo , Mitocondrias/fisiología , Ornitina Descarboxilasa/metabolismo , Inhibidores de la Ornitina Descarboxilasa , Poli(ADP-Ribosa) Polimerasas/metabolismo , Transporte de Proteínas , Proteínas/genética , Proteínas/metabolismo , Transfección , Transgenes/genética , Células Tumorales Cultivadas , Proteína X Asociada a bcl-2/metabolismo , Proteína bcl-X/metabolismoRESUMEN
Ornithine decarboxylase (ODC) plays an essential role in various biological functions, including cell proliferation, differentiation and cell death. However, how it prevents the cell apoptotic mechanism is still unclear. Previous studies have demonstrated that decreasing the activity of ODC by difluoromethylornithine (DFMO), an irreversible inhibitor of ODC, causes the accumulation of intracellular reactive oxygen species (ROS) and cell arrest, thus inducing cell death. These findings might indicate how ODC exerts anti-oxidative and anti-apoptotic effects. In our study, tumor necrosis factor alpha (TNF-alpha) induced apoptosis in HL-60 and Jurkat T cells. The kinetic studies revealed that the TNF-alpha -induced apoptotic process included intracellular ROS generation (as early as 1 h after treatment), the activation of caspase 8 (3 h), the cleavage of Bid (3 h) and the disruption of mitochondrial membrane potential (Delta psi(m)) (6 h). Furthermore, ROS scavengers, such as glutathione (GSH) and catalase, maintained Delta psi(m) and prevented apoptosis upon treatment. Putrescine and overexpression of ODC had similar effects as ROS scavengers in decreasing intracellular ROS and preventing the disruption of Delta psi(m) and apoptosis. Inhibition of ODC by DFMO in HL-60 cells only could increase ROS generation, but did not disrupt Delta psi(m) or induce apoptosis. However, DFMO enhanced the accumulation of ROS, disruption of Delta psi(m) and apoptosis when cells were treated with TNF-alpha . ODC overexpression avoided the decline of Bcl-2, prevented cytochrome c release from mitochondria and inhibited the activation of caspase 8, 9 and 3. Overexpression of Bcl-2 maintained Delta psi(m) and prevented apoptosis, but could not reduce ROS until four hours after TNF-alpha treatment. According to these data, we suggest that TNF-alpha induces apoptosis mainly by a ROS-dependent, mitochondria-mediated pathway. Furthermore, ODC prevents TNF-alpha -induced apoptosis by decreasing intracellular ROS to avoid Bcl-2 decline, maintain Delta psi(m), prevent cytochrome c release and deactivate the caspase cascade pathway.