RESUMEN
The innate immune system is rapidly activated during myocardial infarction and blockade of extracellular complement system reduces infarct size. Intracellular complement, however, appears to be closely linked to metabolic pathways and its role in ischemia-reperfusion injury is unknown and may be different from complement activation in the circulation. The purpose of the present study was to investigate the role of intracellular complement in isolated, retrogradely buffer-perfused hearts and cardiac cells from adult male wild type mice (WT) and from adult male mice with knockout of complement component 3 (C3KO). Main findings: (i) Intracellular C3 protein was expressed in isolated cardiomyocytes and in whole hearts, (ii) after ischemia-reperfusion injury, C3KO hearts had larger infarct size (32 ± 9% in C3KO vs. 22 ± 7% in WT; p=0.008) and impaired post-ischemic relaxation compared to WT hearts, (iii) C3KO cardiomyocytes had lower basal oxidative respiration compared to WT cardiomyocytes, (iv) blocking mTOR decreased Akt phosphorylation in WT, but not in C3KO cardiomyocytes, (v) after ischemia, WT hearts had higher levels of ATP, but lower levels of both reduced and oxidized nicotinamide adenine dinucleotide (NADH and NAD+, respectively) compared to C3KO hearts. Conclusion: intracellular C3 protected the heart against ischemia-reperfusion injury, possibly due to its role in metabolic pathways important for energy production and cell survival.
Asunto(s)
Infarto del Miocardio , Daño por Reperfusión Miocárdica , Animales , Complemento C3 , Homeostasis , Masculino , Ratones , Infarto del Miocardio/metabolismo , Daño por Reperfusión Miocárdica/metabolismo , Miocitos Cardíacos/metabolismoRESUMEN
Docosahexaenoic acid (DHA) and arachidonic acid (ARA) are present in breast milk and play important roles in early infant development. A supply of these fatty acids in infant formula (typically following breast milk as a model with ARA > DHA) is thought to be important since endogenous synthesis is insufficient to maintain tissue levels equivalent to breast-fed infants. Intervention studies assessing the impact of DHA- and ARA-supplemented formulas have resulted in numerous positive developmental outcomes (closer to breast-fed infants) including measures of specific cognition functions, visual acuity, and immune responses. A critical analysis of outcome assessment tools reveals the essentiality of selecting appropriate, focused techniques in order to provide accurate evaluation of DHA- and ARA-supplemented formulas. Future research directions should encompass in-depth assessment of specific cognitive outcomes, immune function, and disease incidence, as well as sources of experimental variability such as the status of fatty acid desaturase polymorphisms.
Asunto(s)
Ácido Araquidónico/farmacología , Ácidos Docosahexaenoicos/farmacología , Fórmulas Infantiles/química , Fenómenos Fisiológicos Nutricionales del Lactante , Lactancia Materna , Desarrollo Infantil , Cognición/efectos de los fármacos , Cognición/fisiología , delta-5 Desaturasa de Ácido Graso , Ácido Graso Desaturasas/genética , Humanos , Inmunidad/efectos de los fármacos , Inmunidad/fisiología , Lactante , Recién Nacido , Leche Humana/química , Polimorfismo Genético , Visión Ocular/efectos de los fármacos , Visión Ocular/fisiologíaRESUMEN
BACKGROUND AND PURPOSE: Defects of coenzyme Q10 (CoQ10) metabolism cause a variety of disorders ranging from isolated myopathy to multisystem involvement. ADCK3 is one of several genes associated with CoQ10 deficiency that presents with progressive cerebellar ataxia, epilepsy, migraine and psychiatric disorders. Diagnosis is challenging due to the wide clinical spectrum and overlap with other mitochondrial disorders. METHODS: A detailed description of three new patients and one previously reported patient from three Norwegian families with novel and known ADCK3 mutations is provided focusing on the epileptic semiology and response to treatment. Mutations were identified by whole exome sequencing and in two measurement of skeletal muscle CoQ10 was performed. RESULTS: All four patients presented with childhood-onset epilepsy and progressive cerebellar ataxia. Three patients had epilepsia partialis continua and stroke-like episodes affecting the posterior brain. Electroencephalography showed focal epileptic activity in the occipital and temporal lobes. Genetic investigation revealed ADCK3 mutations in all patients including a novel change in exon 15: c.T1732G, p.F578V. There was no apparent genotype-phenotype correlation. CONCLUSION: ADCK3 mutations can cause a combination of progressive ataxia and acute epileptic encephalopathy with stroke-like episodes. The clinical, radiological and electrophysiological features of this disorder mimic the phenotype of polymerase gamma (POLG) related encephalopathy and it is therefore suggested that ADCK3 mutations be considered in the differential diagnosis of mitochondrial encephalopathy with POLG-like features.
Asunto(s)
Ataxia/diagnóstico , Ataxia Cerebelosa/diagnóstico , Epilepsia/diagnóstico , Enfermedades Mitocondriales/diagnóstico , Encefalomiopatías Mitocondriales/diagnóstico , Proteínas Mitocondriales/genética , Debilidad Muscular/diagnóstico , Mutación , Ubiquinona/deficiencia , Adulto , Ataxia/genética , Ataxia Cerebelosa/genética , Diagnóstico Diferencial , Epilepsia/genética , Femenino , Humanos , Masculino , Enfermedades Mitocondriales/genética , Debilidad Muscular/genética , Fenotipo , Ubiquinona/genética , Adulto JovenRESUMEN
OBJECTIVE: Primary hyperparathyroidism (PHPT) has been associated with low-grade inflammation and increased risk of cardiovascular disease (CVD). The aim of the study was to investigate systemic levels of pro-inflammatory proteins that previously have not been examined in patients with PHPT. The selection of the pro-inflammatory biomarkers included in this study, MMP9, S100A4, S100A8/A9 and the receptors sCD14 and RAGE, was based on a previous microarray screen of mRNAs in adipose tissue from PHPT patients. DESIGN: A prospective study was conducted on a total of 57 patients with PHPT and a control group of 20 healthy blood donors. METHODS: PHPT patients with normalisation of serum calcium levels after parathyroidectomy were followed for 6 months. Forty-two patients participated in the longitudinal study, in which blood samples were taken at inclusion, and 1, 3 and 6 months after surgery. RESULTS: We observed increased serum levels of MMP9 (P=0.029), S100A4 (P<0.001) and sCD14 (P=0.002) in the 57 patients with PHPT compared to the control-group. During 6 months of follow up, S100A4 (P=0.022) and sCD14 (0.002) decreased significantly, while serum levels of MMP9 increased (P=0.025). CONCLUSIONS: The results demonstrate an increased inflammatory response in PHPT patients shown by elevated MMP9, S100A4 and sCD14 at inclusion. During the 6 months of follow-up, MMP9 increased further, possibly due to the tissue repair process after parathyroidectomy. S100A4 and sCD14 decreased after surgery demonstrating a partial reversal of the systemic inflammation.
Asunto(s)
Biomarcadores/sangre , Hiperparatiroidismo Primario/sangre , Inflamación/sangre , Proteína C-Reactiva/análisis , Estudios de Casos y Controles , Femenino , Estudios de Seguimiento , Humanos , Hiperparatiroidismo Primario/complicaciones , Hiperparatiroidismo Primario/cirugía , Inflamación/etiología , Receptores de Lipopolisacáridos/sangre , Estudios Longitudinales , Masculino , Metaloproteinasa 9 de la Matriz/sangre , Análisis por Micromatrices , Persona de Mediana Edad , Paratiroidectomía , Estudios Prospectivos , Receptor para Productos Finales de Glicación Avanzada , Receptores Inmunológicos/sangre , Proteína de Unión al Calcio S100A4 , Proteínas S100/sangreRESUMEN
The velocity of a swimmer is determined by biomechanical and bioenergetics factors. However, little is known about the effect of ankle flexibility on dolphin kick performance. Next to this, scientific evidence is lacking concerning the influence of ankle muscle strength. Therefore, the aim of this study was to investigate the effect of ankle flexibility and muscle strength on dolphin kick performance in competitive swimmers. Ankle range of motion (ROM) and ankle muscle strength were measured in 26 healthy competitive swimmers. The effect of both was assessed on the swimmer's velocity and lower extremity joint angles during three maximal dolphin kick trials. Additionally, the effect of a flexibility restriction by a tape on the dolphin kick performance was assessed. Correlations were calculated between the flexibility, muscle strength and dolphin kick performance and differences were investigated between the unrestricted and restricted condition. Muscle strength of dorsal flexors and internal rotators were positively significantly correlated with the velocity. Active and passive plantar flexion ROM and internal rotation ROM were not significantly correlated. A plantar flexion-internal rotation restriction during the dolphin kick showed a significant decrease in velocity. This restriction was associated with a changed movement pattern in the knee towards more flexion. The results suggest that dolphin kick velocity might be enhanced by ankle muscle strength exercises and that subjects with a restricted ankle flexibility might profit from a flexibility program.
Asunto(s)
Tobillo/fisiología , Atletas , Destreza Motora/fisiología , Fuerza Muscular/fisiología , Músculo Esquelético/fisiología , Natación/fisiología , Adolescente , Articulación del Tobillo/fisiología , Rendimiento Atlético , Fenómenos Biomecánicos , Niño , Femenino , Humanos , Rodilla/fisiología , Articulación de la Rodilla/fisiología , Masculino , Movimiento , Rango del Movimiento Articular/fisiología , Adulto JovenRESUMEN
The conversion of the plant-derived omega-3 (n-3) α-linolenic acid (ALA, 18:3n-3) to the long-chain eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) can be increased by ALA sufficient diets compared to ALA deficient diets. Diets containing ALA above an optimal level result in no further increase in DHA levels in animals and humans. The present study evaluates means of maximizing plasma DHA accumulation by systematically varying both linoleic acid (LA, 18:2n-6) and ALA dietary level. Weanling rats were fed one of 54 diets for three weeks. The diets varied in the percentage of energy (en%) of LA (0.07-17.1 en%) and ALA (0.02-12.1 en%) by manipulating both the fat content and the balance of vegetable oils. The peak of plasma phospholipid DHA (>8% total fatty acids) was attained as a result of feeding a narrow dietary range of 1-3 en% ALA and 1-2 en% LA but was suppressed to basal levels (â¼2% total fatty acids) at dietary intakes of total polyunsaturated fatty acids (PUFA) above 3 en%. We conclude it is possible to enhance the DHA status of rats fed diets containing ALA as the only source of n-3 fatty acids but only when the level of dietary PUFA is low (<3 en%).
Asunto(s)
Dieta Alta en Grasa/efectos adversos , Ácidos Docosahexaenoicos/metabolismo , Ácidos Grasos Esenciales/metabolismo , Ácidos Grasos Insaturados/administración & dosificación , Ácido alfa-Linolénico/metabolismo , Algoritmos , Animales , Dieta con Restricción de Grasas , Ácidos Docosahexaenoicos/sangre , Ácido Eicosapentaenoico/sangre , Ácido Eicosapentaenoico/metabolismo , Ácidos Grasos Esenciales/sangre , Ácidos Grasos Esenciales/deficiencia , Ácidos Grasos Omega-6/efectos adversos , Ácidos Grasos Omega-6/sangre , Ácidos Grasos Omega-6/química , Ácidos Grasos Omega-6/metabolismo , Ácidos Grasos Insaturados/efectos adversos , Ácidos Grasos Insaturados/análisis , Ácidos Grasos Insaturados/sangre , Ácido Linoleico/administración & dosificación , Ácido Linoleico/efectos adversos , Ácido Linoleico/sangre , Ácido Linoleico/metabolismo , Aceite de Linaza/administración & dosificación , Aceite de Linaza/química , Aceite de Linaza/metabolismo , Masculino , Fosfolípidos/sangre , Fosfolípidos/química , Fosfolípidos/metabolismo , Aceites de Plantas/administración & dosificación , Aceites de Plantas/efectos adversos , Aceites de Plantas/química , Aceites de Plantas/metabolismo , Ratas , Ratas Wistar , Aceite de Cártamo/administración & dosificación , Aceite de Cártamo/efectos adversos , Aceite de Cártamo/química , Aceite de Cártamo/metabolismo , Aceite de Girasol , Destete , Ácido alfa-Linolénico/administración & dosificación , Ácido alfa-Linolénico/análisis , Ácido alfa-Linolénico/sangreRESUMEN
OBJECTIVES: Primary hyperparathyroidism (PHPT) has been associated with low-grade inflammation and elevated risk of cardiovascular disease (CVD). In inflammatory conditions, interferon-γ (IFN-γ) activity is enhanced and a decreased circulating concentration of vitamin B6 is often observed. Such changes in IFN-γ activity or vitamin B6 levels have been associated with increased incidence of CVD. The aim of the study was to investigate systemic markers of IFN-γ-mediated immune activation, such as neopterin, the kynurenine-to-tryptophan ratio (KTR) and kynurenine pathway metabolites, as well as B6 vitamers in patients with PHPT. DESIGN/SUBJECTS: A total of 57 patients with PHPT and a control group of 20 healthy blood donors were included in this study. PHPT patients who responded positively to parathyroidectomy were followed for 6 months. Forty-three patients participated in the longitudinal study in which blood samples were taken at inclusion and 1, 3 and 6 months after surgery. RESULTS: Plasma concentrations of the B6 vitamers pyridoxal 5'-phosphate (PLP) (P = 0.007) and pyridoxal (P = 0.013) were significantly lower in the patient group compared to healthy control subjects. An increase in the KTR indicated that the kynurenine pathway of tryptophan metabolism was altered in PHPT patients (P = 0.015). During the initial 6 months after surgery, levels of PLP (P < 0.001) and anthranilic acid (P < 0.001) increased significantly, whereas neopterin decreased (P = 0.018). CONCLUSIONS: The results of this study demonstrate altered levels of vitamin B6 and the KTR in PHPT patients, both of which may reflect cellular immune activation. These abnormalities should be considered in relation to the increased risk of CVD previously observed in patients with PHPT.
Asunto(s)
Hiperparatiroidismo Primario , Quinurenina/metabolismo , Paratiroidectomía/métodos , Triptófano/metabolismo , Vitamina B 6 , Anciano , Biomarcadores , Enfermedades Cardiovasculares/etiología , Enfermedades Cardiovasculares/metabolismo , Enfermedades Cardiovasculares/prevención & control , Femenino , Humanos , Hiperparatiroidismo Primario/complicaciones , Hiperparatiroidismo Primario/metabolismo , Hiperparatiroidismo Primario/cirugía , Inmunidad Celular , Factores Inmunológicos , Inflamación/metabolismo , Interferón gamma/metabolismo , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Monitorización Inmunológica/métodos , Neopterin/metabolismo , Cuidados Posoperatorios/métodos , Factores de Riesgo , Vitamina B 6/sangre , Vitamina B 6/metabolismo , ortoaminobenzoatos/metabolismoRESUMEN
BACKGROUND: Studies have shown that Ki-67 response after short-term neoadjuvant aromatase inhibitors may predict recurrence in postmenopausal breast cancer, whereas its prognostic effect in premenopausal women is unknown. PATIENTS AND METHODS: We compared the prognostic and predictive value of baseline and post-treatment Ki-67 in 120 pre- and postmenopausal women with early-stage estrogen receptor-positive breast cancer who participated in a 4-week presurgical trial of tamoxifen. RESULTS: After 7.2 years of follow-up, women with post-treatment Ki-67 in the second (14%-19%), third (20%-29%) and top (≥30%) quartiles had a recurrence hazard ratio of 2.92 [95% confidence interval (CI) 0.95-8.96], 4.37 (1.56-12.25) and 6.05 (2.07-17.65), respectively, as compared with those in the bottom quartile (<14%) (P-trend = 0.001). The risk of invasive disease recurrence was 2.2% (95% CI 0.9-5.0) per point increase in baseline Ki-67 (P-trend = 0.076) and 5.0% (95% CI 2.3-7.7) per point increase in post-tamoxifen Ki-67 (P-trend < 0.001). The risk of death was 5.5 (95% CI 1.26-23.16) times higher in patients with post-drug Ki-67 ≥20% than in those with Ki-67 <20% (P-trend = 0.006). CONCLUSIONS: Ki-67 response after short-term neoadjuvant tamoxifen is a good predictor of recurrence-free survival and overall survival, further supporting its use as surrogate biomarker to personalize adjuvant treatment and to screen novel drugs cost-effectively.
Asunto(s)
Antineoplásicos Hormonales/uso terapéutico , Neoplasias de la Mama/patología , Antígeno Ki-67/metabolismo , Neoplasias Hormono-Dependientes/patología , Receptores de Estrógenos/metabolismo , Tamoxifeno/uso terapéutico , Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/cirugía , Método Doble Ciego , Femenino , Estudios de Seguimiento , Humanos , Estimación de Kaplan-Meier , Persona de Mediana Edad , Terapia Neoadyuvante , Invasividad Neoplásica , Recurrencia Local de Neoplasia , Neoplasias Hormono-Dependientes/tratamiento farmacológico , Neoplasias Hormono-Dependientes/cirugía , Valor Predictivo de las Pruebas , Pronóstico , Modelos de Riesgos Proporcionales , Receptor ErbB-2/metabolismo , Receptores de Progesterona/metabolismo , Coloración y Etiquetado , Carga Tumoral/efectos de los fármacosRESUMEN
BACKGROUND: Acquired resistance to endocrine therapy in breast cancer is poorly understood. Characterisation of the molecular response to aromatase inhibitors in breast cancer tissue may provide important information regarding development of oestrogen hypersensitivity. METHODS: We examined the expression levels of nuclear receptor co-regulators, the orphan nuclear receptor liver receptor homologue-1 and HER-2/neu growth factor receptor using real-time RT-PCR before and after 13-16 weeks of primary medical treatment with the aromatase inhibitors anastrozole or letrozole. RESULTS: mRNA expression of the steroid receptor co-activator 1 (SRC-1) and peroxisome-proliferator-activated receptor gamma co-activator-1alpha (PGC-1alpha) was correlated (P=0.002), and both co-activators increased during treatment in the patient group as a whole (P=0.008 and P=0.032, respectively), as well as in the subgroup of patients achieving an objective treatment response (P=0.002 and P=0.006). Although we recorded no significant change in SRC-3/amplified in breast cancer 1 level, the expression correlated positively to the change of SRC-1 (P=0.002). Notably, we recorded an increase in HER-2/neu levels during therapy in the total patient group (18 out of 26; P=0.016), but in particular among responders (15 out of 21; P=0.008). CONCLUSION: Our results show an upregulation of co-activator mRNA and HER-2/neu during treatment with aromatase inhibitors. These mechanisms may represent an early adaption of the breast cancer cells to oestrogen deprivation in vivo.
Asunto(s)
Inhibidores de la Aromatasa/uso terapéutico , Neoplasias de la Mama/tratamiento farmacológico , Proteínas de Choque Térmico/genética , Histona Acetiltransferasas/genética , Receptor ErbB-2/genética , Factores de Transcripción/genética , Neoplasias de la Mama/metabolismo , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Terapia Neoadyuvante , Coactivador 1 de Receptor Nuclear , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma , ARN Mensajero/análisis , Receptores Citoplasmáticos y Nucleares/genética , Receptores de Estrógenos/análisisRESUMEN
OBJECTIVE: To evaluate the efficacy and safety of an alpha-lactalbumin-enriched formula with a protein profile and total protein concentration closer to human milk (HM) and lower than conventional formulas. SUBJECTS/METHODS: Two hundred and sixteen healthy, term infants, Asunto(s)
Aminoácidos Esenciales/sangre
, Fórmulas Infantiles/química
, Fenómenos Fisiológicos Nutricionales del Lactante/fisiología
, Recién Nacido/sangre
, Recién Nacido/crecimiento & desarrollo
, Lactalbúmina/administración & dosificación
, Necesidades Nutricionales
, Proteínas en la Dieta/administración & dosificación
, Femenino
, Humanos
, Lactante
, Fórmulas Infantiles/administración & dosificación
, Masculino
, Leche Humana/química
, Estudios Prospectivos
, Aumento de Peso
RESUMEN
BACKGROUND: Tamoxifen is hydroxylated by cytochrome P450 (CYP) 2D6 to the potent metabolites 4-hydroxytamoxifen (4OHtam) and 4-hydroxy-N-demethyltamoxifen (4OHNDtam), which are both conjugated by sulphotransferase (SULT)1A1. Clinical studies indicate that CYP2D6 and SULT1A1 genotypes are predictors for treatment response to tamoxifen. Therefore, we examined the relationship between CYP2D6 genotype, SULT1A1 genotype, SULT1A1 copy number and the pharmacokinetics of tamoxifen. PATIENTS AND METHODS: The serum levels of tamoxifen and metabolites of 151 breast cancer patients were measured by high-pressure liquid chromatography-tandem mass spectrometry. The CYP2D6 and SULT1A1 polymorphisms and SULT1A1 copy number were determined by long PCR, PCR-based restriction fragment length polymorphism, DNA sequencing and fluorescence-based PCR. RESULTS: The levels of 4OHtam, 4OHNDtam and N-demethyltamoxifen were associated with CYP2D6 predicted enzymatic activity (P < 0.05). The SULT1A1 genotype or copy number did not influence the levels of tamoxifen and its metabolites. However, the ratios of N-demethyltamoxifen/tamoxifen and N-dedimethyltamoxifen/N-demethyltamoxifen were related to SULT1A1 genotype. CONCLUSION: CYP2D6 and SULT1A1 genotypes may partly explain the wide inter-individual variations in the serum levels of tamoxifen and its metabolites. We propose that therapeutic drug monitoring should be included in studies linking CYP2D6 and SULT1A1 genotypes to clinical outcome.
Asunto(s)
Antineoplásicos Hormonales/farmacocinética , Arilsulfotransferasa/genética , Neoplasias de la Mama/tratamiento farmacológico , Citocromo P-450 CYP2D6/genética , Moduladores Selectivos de los Receptores de Estrógeno/farmacocinética , Tamoxifeno/farmacocinética , Adulto , Anciano , Anciano de 80 o más Años , Arilsulfotransferasa/metabolismo , Biotransformación/genética , Neoplasias de la Mama/sangre , Neoplasias de la Mama/enzimología , Citocromo P-450 CYP2D6/metabolismo , Femenino , Dosificación de Gen , Frecuencia de los Genes , Genotipo , Humanos , Persona de Mediana Edad , Noruega , Polimorfismo de Longitud del Fragmento de Restricción , Tamoxifeno/análogos & derivados , Tamoxifeno/sangreRESUMEN
Aromatase inhibitors improve relapse-free survival in early breast cancer, but there is concern about possible detrimental effects on bone mineral density (BMD) and plasma lipids. This paper presents the results of a 2-year study evaluating the effects of exemestane versus placebo on BMD, bone markers, plasma lipids and coagulation factors, including a 1-year follow-up after termination of treatment in 147 patients. During treatment, the mean annual rate of loss of BMD in the lumbar spine was 2.17% in the exemestane group versus 1.84% in the placebo group (n.s.) and 2.72% versus 1.48%, respectively, in the femoral neck (P=0.024). A loss of BMD above that expected in both arms of this study could be due to low vitamin D status (88% of all patients had vitamin D levels <30 ng/ml). The changes observed with exemestane were partially reversed during a 1-year follow-up, with no significant difference between the two arms. Similarly, the moderate decrease in high-density lipoprotein (HDL)-cholesterol was reversed. The bone marker values decreased, although a difference at 6 months of follow-up was still recorded, in particular for the markers of bone synthesis.
Asunto(s)
Androstadienos/efectos adversos , Inhibidores de la Aromatasa/efectos adversos , Densidad Ósea/efectos de los fármacos , Neoplasias de la Mama/tratamiento farmacológico , Biomarcadores/metabolismo , Factores de Coagulación Sanguínea/metabolismo , Remodelación Ósea , Huesos/metabolismo , Neoplasias de la Mama/metabolismo , Femenino , Estudios de Seguimiento , Hormonas Esteroides Gonadales/sangre , Homocisteína/sangre , Humanos , Lípidos/sangre , Persona de Mediana Edad , Posmenopausia , Vitamina D/sangre , Privación de TratamientoRESUMEN
We have developed a method for the determination of tamoxifen (tam) and its metabolites 4-hydroxytamoxifen (4OHtam), N-demethyltamoxifen (NDtam), N-dedimethyltamoxifen (NDDtam), tamoxifen-N-oxide (tamNox), and 4-hydroxy-N-demethyltamoxifen (4OHNDtam) in 50 microl human serum. Serum proteins were precipitated with acetonitrile. Deuterated-tamoxifen (D5 tam) was added as internal standard. Sample supernatant was injected into an on-line reversed-phase extraction column coupled with a C18 analytical column and analytes were detected by tandem mass spectrometry. The lower limits of quantification were 0.25 ng/mL for 4OHtam, NDtam and tam, 1.0 ng/mL for NDDtam and tamNox. Ranges of within- and between-day variation were 2.9-15.4% and 4.4-12.9%, respectively.
Asunto(s)
Cromatografía Liquida/métodos , Espectrometría de Masas/métodos , Tamoxifeno/sangre , Neoplasias de la Mama/tratamiento farmacológico , Fraccionamiento Químico/métodos , Humanos , Sensibilidad y Especificidad , Tamoxifeno/análogos & derivados , Tamoxifeno/metabolismo , Tamoxifeno/uso terapéuticoRESUMEN
Toll-like receptor 2 (TLR2) stimulation in monocytes may contribute to enhanced inflammation and viral replication in HIV infection. In the present study we examined if TLR2 stimulation could modulate chemokine responses in peripheral blood mononuclear cells (PBMC) from HIV-infected patients and healthy controls. Our main findings were, with similar qualitative patterns in both healthy controls and HIV-infected patients: (1) TLR2 stimulation induced up-regulation of several chemokines at the mRNA level as well as increased protein levels of macrophage inflammatory protein (MIP)-1alpha, interleukin (IL)-8 and regulated on activation, normal T cell expressed and secreted (RANTES); (2) TLR2 stimulation induced enhanced protein expression of CCR5 (a receptor for MIP-1alpha and RANTES) on monocytes; (3) In vitro stimulation with RANTES induced release of MIP-1alpha, MCP-1, IL-8 and interferon-gamma from PBMC. While increased levels of beta-chemokines possibly have antiviral effects, TLR2 stimulation may also promote a chemokine-driven inflammatory loop, potentially contributing to the immunopathogenesis of HIV infection.
Asunto(s)
Quimiocinas/inmunología , Infecciones por VIH/inmunología , Leucocitos Mononucleares/inmunología , Glicoproteínas de Membrana/inmunología , Receptores de Superficie Celular/inmunología , Adulto , Terapia Antirretroviral Altamente Activa/métodos , Quimiocina CCL2/inmunología , Quimiocina CCL3 , Quimiocina CCL4 , Quimiocina CCL5/análisis , Femenino , Infecciones por VIH/tratamiento farmacológico , Humanos , Interferón gamma/inmunología , Interleucina-8/análisis , Interleucina-8/inmunología , Proteínas Inflamatorias de Macrófagos/análisis , Proteínas Inflamatorias de Macrófagos/inmunología , Masculino , Persona de Mediana Edad , ARN Mensajero/análisis , ARN Viral/análisis , Receptores CCR5/análisis , Proteínas Recombinantes/inmunología , Receptor Toll-Like 2 , Receptores Toll-Like , Regulación hacia Arriba/inmunologíaAsunto(s)
Lactoferrina/análisis , Leche Humana/química , Adolescente , Adulto , Análisis de Varianza , Australia , Canadá , China , Cromatografía Líquida de Alta Presión/métodos , Femenino , Humanos , Recién Nacido , Japón , México , Nitrógeno/análisis , Filipinas , Reino Unido , Estados UnidosRESUMEN
BACKGROUND: From the deductive point of view, neurotransmitter receptors can be divided into categories such as cholinergic (muscarinic, nicotinic), adrenergic (alpha- and beta-), dopaminergic, serotoninergic (5-HT1 approximately 5-HT5), and histaminergic (H1 and H2). Selective agonists and antagonists of each receptor subtype can have specific useful therapeutic applications. For understanding the molecular mechanisms of action, an inductive method of analysis is useful. OBJECTIVE: The aim of the present study is to examine the structure-activity relationships of agents acting on G-protein coupled receptors. METHOD: Representative sets of G-PCR agonists and antagonists were identified from the literature and Medline [P.M. Walsh (2003) Physicians' Desk Reference; M.J. O'Neil (2001) The Merck Index]. The molecular weight (MW), calculated logarithm of octanol/water partition coefficient (C log P) and molar refraction (CMR), dipole moment (DM), E(lumo) (the energy of the lowest unoccupied molecular orbital, a measure of the electron affinity of a molecule and its reactivity as an electrophile), E(homo) (the energy of the highest occupied molecular orbital, related to the ionization potential of a molecule, and its reactivity as a nucleophile), and the total number of hydrogen bonds (H(b)) (donors and receptors), were chosen as molecular descriptors for SAR analyses. RESULTS: The data suggest that not only do neurotransmitters share common structural features but their receptors belong to the same ensemble of G-protein coupled receptor with seven to eight transmembrane domains with their resultant dipoles in an antiparallel configuration. Moreover, the analysis indicates that the receptor exists in a dynamic equilibrium between the closed state and the open state. The energy needed to open the closed state is provided by the hydrolysis of GTP. A composite 3-D parameter frame setting of all the neurotransmitter agonists and antagonists are presented using MW, Hb and mu as independent variables. CONCLUSION: It appears that all neurotransmitters examined in this study operate by a similar mechanism with the G-protein coupled receptors.
Asunto(s)
Neurotransmisores/antagonistas & inhibidores , Receptores Acoplados a Proteínas G/efectos de los fármacos , Receptores Acoplados a Proteínas G/fisiología , Relación Estructura-Actividad , Agonistas Adrenérgicos/química , Agonistas Adrenérgicos/clasificación , Antagonistas Adrenérgicos/química , Antagonistas Adrenérgicos/clasificación , Fenómenos Químicos , Química Física , Agonistas Colinérgicos/química , Agonistas Colinérgicos/clasificación , Antagonistas Colinérgicos/química , Antagonistas Colinérgicos/clasificación , Agonistas de Dopamina/química , Agonistas de Dopamina/clasificación , Agonistas de Dopamina/farmacología , Antagonistas de Dopamina/química , Antagonistas de Dopamina/clasificación , Antagonistas de Dopamina/farmacología , Agonistas de los Receptores Histamínicos/química , Agonistas de los Receptores Histamínicos/clasificación , Agonistas de los Receptores Histamínicos/farmacología , Antagonistas de los Receptores Histamínicos/química , Antagonistas de los Receptores Histamínicos/clasificación , Antagonistas de los Receptores Histamínicos/farmacología , Modelos Biológicos , Estructura Molecular , Neurotransmisores/agonistas , Neurotransmisores/química , Receptores Adrenérgicos/clasificación , Receptores Adrenérgicos/efectos de los fármacos , Receptores Adrenérgicos/fisiología , Receptores Colinérgicos/clasificación , Receptores Colinérgicos/efectos de los fármacos , Receptores Colinérgicos/fisiología , Receptores Histamínicos/clasificación , Receptores Histamínicos/efectos de los fármacos , Receptores Histamínicos/fisiología , Antagonistas de la Serotonina/química , Antagonistas de la Serotonina/clasificación , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/química , Agonistas de Receptores de Serotonina/clasificación , Agonistas de Receptores de Serotonina/farmacologíaRESUMEN
Earlier findings have suggested that the balance between interleukin-10 and tumor necrosis factor alpha levels in serum may influence the outcome of cytomegalovirus infection in renal transplant recipients. Therefore, the aim of the present study was to investigate whether human cytomegalovirus induces interleukin-10 production in macrophages. Experiments using human cytomegalovirus (strain 2006), ultraviolet-inactivated cytomegalovirus, and mock-infected differentiated THP-1 cells with or without ganciclovir or monoclonal anti-tumor necrosis factor alpha antibodies were performed. Cytomegalovirus-infected cells produced significantly higher levels of human interleukin-10 mRNA and interleukin-10 than ultraviolet-inactivated cytomegalovirus or mock-infected cells. The addition of ganciclovir had little effect on interleukin-10 production. Anti-tumor necrosis factor alpha antibodies appeared to reduce the interleukin-10 levels. In conclusion, human cytomegalovirus infection of macrophages induces production of human interleukin-10. This requires viral entry, but not full viral replication.
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Infecciones por Citomegalovirus/fisiopatología , Citomegalovirus/inmunología , Citomegalovirus/fisiología , Interleucina-10/biosíntesis , Macrófagos/fisiología , Factor de Necrosis Tumoral alfa/biosíntesis , Biomarcadores , Células Cultivadas , Humanos , Probabilidad , Pronóstico , ARN Mensajero/análisis , Sensibilidad y Especificidad , Estadísticas no ParamétricasRESUMEN
OBJECTIVES: To investigate plasma total homocysteine levels and its relation to B-vitamins and smoking in Graves' disease before and after antithyroid therapy. DESIGN: A longitudinal study taking place at four hospitals in Norway. METHODS AND SUBJECTS: Plasma total homocysteine, serum folate, serum cobalamin and riboflavin, flavin mononucleotide and flavin adenine dinucleotide in plasma were investigated in 182 patients with hyperthyroidism before treatment. The same parameters were reinvestigated in 112 of these patients after attaining euthyroid state. RESULTS: In hyperthyroidism, plasma total homocysteine was low, and inversely related to folate, cobalamin and riboflavin, and positively related to serum creatinine and age. Following antithyroid therapy, total homocysteine increased and the concentration of folate, cobalamin, riboflavin, flavin mononucleotide and flavin adenine dinucleotide decreased significantly. The most pronounced reduction (35%) was observed for flavin mononucleotide. In the hyperthyroid state, smokers had lower levels of folate and flavin mononucleotide than non-smokers. After restoration of euthyroidism, both folate and riboflavin were significantly lower in smokers than non-smokers. Plasma total homocysteine increased according to decreasing quartiles of B-vitamins. For riboflavin, this relation was confined to smokers. CONCLUSION: Plasma total homocysteine changes according to thyroid status. These changes may be partly attributable to altered folate, cobalamin but also riboflavin status, particularly in smokers.
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Antitiroideos/uso terapéutico , Enfermedad de Graves/sangre , Homocisteína/sangre , Fumar/sangre , Complejo Vitamínico B/sangre , Adolescente , Adulto , Anciano , Carbimazol/uso terapéutico , Creatinina/sangre , Femenino , Ácido Fólico/sangre , Enfermedad de Graves/tratamiento farmacológico , Humanos , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Análisis de Regresión , Riboflavina/sangre , Vitamina B 12/sangreRESUMEN
The use of tamoxifen as a breast cancer preventive agent may be contraindicated by an increased risk of endometrial cancer and venous thromboembolic events, particularly in postmenopausal women. Since these estrogenic effects may be dose-related, a dose reduction may reduce toxicity. We have recently shown a comparable activity of lower doses of tamoxifen on putative surrogate biomarkers of cardiovascular disease and breast cancer. To provide further insight into the effect of tamoxifen at low doses on the cardiovascular system, we compared the effect of three different doses on circulating levels of C-reactive protein (CRP), an independent risk marker for cardiovascular disease (CVD), which was lowered by tamoxifen at the standard dose of 20 mg day-1 in previous studies. We compared the changes in CRP after 2 months of either placebo (n = 24), or tamoxifen 10 mg alternate daily (n = 26), or 10 mg day-1 (n = 22), or 20 mg day-1 (n = 19) in healthy women aged 35-70 years. The median percent change was -2.2% (95% CI, -23.3 to 42.8) with placebo, -39.1 (95% CI, -59.9 to -28.7) with 10 mg alternate daily, -56.9% (95% CI, -68.6 to -38.4) with 10 mg day-1 and -42.9% (95% CI, -62.6 to 1.6) with 20 mg day-1 (P = 0.291 for the linear dose-response trend). Similar results were obtained when the data were classified according to blood tamoxifen concentrations, with a median reduction of 47% (95% CI, 65.5-36.3) for women with low tamoxifen concentrations (< 30 ng mL-1). We conclude that tamoxifen at low doses is able to lower ultrasensitive CRP and that this might be associated with a beneficial effect on CVD.
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Proteína C-Reactiva/biosíntesis , Tamoxifeno/uso terapéutico , Adulto , Anciano , Neoplasias de la Mama/prevención & control , Enfermedades Cardiovasculares/etiología , Enfermedades Cardiovasculares/prevención & control , Estudios de Cohortes , Relación Dosis-Respuesta a Droga , Antagonistas de Estrógenos/uso terapéutico , Estrógenos/metabolismo , Femenino , Humanos , Histerectomía , Persona de Mediana Edad , Placebos , Factores de RiesgoRESUMEN
OBJECTIVE: A large number of structurally and functionally diverse compounds act as substrates or modulators of p-glycoprotein (p-gp). Some of them possess multiple drug resistance (MDR)-reversing activity, but only a small number of them have entered clinical study. In order to uncover the factors which exert a significant impact on the interaction between substrates/modulators and p-gp, we have performed structure-activity relationship (SAR) analyses, including molecular modelling, two-dimensional (2D) and three-dimensional (3D) parameter-frame-setting analysis, quantitative structure activity relationship (QSAR) analysis among substrates/modulators, as well as clinically promising MDR-reversing agents. METHODS: The physicochemical parameters C log P, CMR and all regression equations were derived by using C log P version 4.0 and the latest CQSAR software, respectively. Molecular modelling and all other parameter calculations were performed by using HyperChem version 5.0 program, after geometry optimization and energy minimization using the AM1 semiempirical method. RESULTS: SAR analyses indicate that MDR reversal activity is correlated with the lipophilicity (C log P), molecular weight (log Mw), longest chain (Nlc) of the molecule and the energy of the highest occupied orbital (Ehomo). In addition, the presence of a basic tertiary nitrogen atom in the structure is also an important contributor to p-gp inhibitory activity. Some separation in space is achieved for different subsets of p-gp substrates and inhibitors using Nlc, C log P and Ehomo as three independent parameters in the 3D-parameter-frame setting. CONCLUSION: A highly effective p-gp modulator candidate should possess a log P value of 2.92 or higher, 18-atom-long or longer molecular axis, and a high Ehomo value, as well as at least one tertiary basic nitrogen atom. The results obtained may be useful in explaining drug-p-gp interactions for different compounds, including drug interactions and the development of new MDR chemosensitizers.