RESUMEN
Various complement-mediated renal disorders are treated currently with the complement inhibitor eculizumab. By blocking the cleavage of C5, this monoclonal antibody prevents cell damage caused by complement-mediated inflammation. We included 23 patients with atypical haemolytic uraemic syndrome (aHUS, n = 12), C3 glomerulopathies (C3G, n = 9) and acute antibody-mediated renal graft rejection (AMR, n = 2), treated with eculizumab in 12 hospitals in Germany. We explored the course of complement activation biomarkers and the benefit of therapeutic drug monitoring of eculizumab. Complement activation was assessed by analysing the haemolytic complement function of the classical (CH50) and the alternative pathway (APH50), C3 and the activation products C3d, C5a and sC5b-9 prior to, 3 and 6 months after eculizumab treatment. Eculizumab concentrations were determined by a newly established specific enzyme-linked immunosorbent assay (ELISA). Serum eculizumab concentrations up to 1082 µg/ml point to drug accumulation, especially in paediatric patients. Loss of the therapeutic antibody via urine with concentrations up to 56 µg/ml correlated with proteinuria. In aHUS patients, effective complement inhibition was demonstrated by significant reductions of CH50, APH50, C3d and sC5b-9 levels, whereas C5a levels were only reduced significantly after 6 months' treatment. C3G patients presented increased C3d and consistently low C3 levels, reflecting ongoing complement activation and consumption at the C3 level, despite eculizumab treatment. A comprehensive complement analysis together with drug monitoring is required to distinguish mode of complement activation and efficacy of eculizumab treatment in distinct renal disorders. Accumulation of the anti-C5 antibody points to the need for a patient-orientated tailored therapy.
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Anticuerpos Monoclonales Humanizados/uso terapéutico , Síndrome Hemolítico Urémico Atípico/tratamiento farmacológico , Complemento C3/inmunología , Glomerulonefritis Membranosa/tratamiento farmacológico , Rechazo de Injerto/prevención & control , Inmunosupresores/uso terapéutico , Trasplante de Riñón , Adolescente , Adulto , Citotoxicidad Celular Dependiente de Anticuerpos/efectos de los fármacos , Biomarcadores/metabolismo , Niño , Preescolar , Activación de Complemento/efectos de los fármacos , Complemento C5/inmunología , Femenino , Humanos , Lactante , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
INTRODUCTION: Nonobstructive cholestasis after pediatric liver transplantation is a common diagnostic and therapeutic dilemma. We describe a girl with neonatal cholestasis because of progressive familial intrahepatic cholestasis 2 (PFIC-2) and presence of a homozygous splice site mutation in the ABCB11 gene. Liver transplantation was performed because of end-stage liver disease at the age of 6. Cholestasis with normal gamma-glutamyl transferase (GGT) developed 8 years after liver transplantation. A liver biopsy showed canalicular cholestasis and giant cell hepatitis without evidence of rejection, mimicking PFIC-2. Immunofluorescence staining of normal human liver sections with patient's serum revealed reactivity toward a canalicular epitope, which could be identified as bile salt export pump (BSEP) using BSEP-yellow fluorescent protein (YFP) transfected cells. Our patient developed a recurrence of a PFIC-2 phenotype due to production of antibodies against BSEP (alloimmune BSEP disease [AIBD]). Intensification of immunosuppressive therapy as well as antibody treatment with plasmapheresis and Rituximab were initiated, leading to stabilization of the clinical condition and depletion of anti-BSEP antibodies in serum. However, after 1 year liver transplantation was necessary again because of end-stage liver insufficiency. Afterward, immunomodulatory treatment consisted of tacrolimus, mycophenolate mofetil, prednisone, immunoadsorption, and high-dose immunoglobulin therapy (1 g/kg/d). CONCLUSION: Cholestasis after liver transplantation may indicate an AIBD with a PFIC-2 phenotype. Besides enhancement of immunosuppressive therapy, an antibody depletion with plasmapheresis, immunoadsorption, immunoglobulins, and B-cell depletion represents a therapeutic option.
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Colestasis Intrahepática/inmunología , Enfermedad Hepática en Estado Terminal/inmunología , Inmunosupresores/uso terapéutico , Trasplante de Hígado/efectos adversos , Plasmaféresis/métodos , Miembro 11 de la Subfamilia B de Transportador de Casetes de Unión al ATP/genética , Miembro 11 de la Subfamilia B de Transportador de Casetes de Unión al ATP/inmunología , Adolescente , Anticuerpos/sangre , Anticuerpos/inmunología , Linfocitos B/inmunología , Niño , Colestasis Intrahepática/complicaciones , Colestasis Intrahepática/genética , Diagnóstico Diferencial , Enfermedad Hepática en Estado Terminal/genética , Enfermedad Hepática en Estado Terminal/cirugía , Epítopos , Femenino , Humanos , Factores Inmunológicos/uso terapéutico , Terapia de Inmunosupresión/métodos , Mutación , Fenotipo , Periodo Posoperatorio , Recurrencia , Reoperación/métodos , Rituximab/uso terapéutico , Resultado del TratamientoRESUMEN
BACKGROUND: Recent evidence indicates that certain genotypes of beta(2)-adrenoceptors (AR) may indicate an increased risk of cardiovascular disease or an increased rate of disease progression. Of particular importance, the Thr164Ile polymorphism, which is found in approximately 4% of humans, shows decreased receptor signaling, blunted cardiac response when expressed in transgenic mice, and is associated with a decreased survival rate in patients with congestive heart failure. METHODS AND RESULTS: In this study, we compared functional activity, ie, chronotropic (heart rate increases) and inotropic (duration of the electromechanical systole) responses to intravenously administered terbutaline, in 6 subjects (4 women and 2 men) who were heterozygous for Thr164Ile with the responses in 12 volunteers (6 women and 6 men) who were homozygous for wild-type (WT) beta(2)-AR (ie, Arg16, Gln27, and Thr164). The beta(2)AR polymorphism significantly affected the dose-response curves for terbutaline-induced inotropic and chronotropic responses: compared with WT individuals, subjects with the Thr164Ile receptor had substantial blunting in maximal increases in heart rate (WT, 29.7+/-3.9 beats/min; Ile164, 20.7+/-1.9 beats/min; P:=0.016) and a shortening of the duration of electromechanical systole (WT, 51.9+/-4.5 ms; Ile164, 37.9+/-4.6 ms; P:=0.02). CONCLUSIONS: These data show that humans with the Ile164 genotype show blunted cardiac beta(2)-AR responsiveness, which may help explain the decreased survival of patients with this genotype in the setting of congestive heart failure.
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Agonistas Adrenérgicos beta/farmacología , Insuficiencia Cardíaca/genética , Receptores Adrenérgicos beta 2/genética , Terbutalina/farmacología , Adulto , Sustitución de Aminoácidos , Femenino , Pruebas Genéticas , Genotipo , Insuficiencia Cardíaca/metabolismo , Insuficiencia Cardíaca/fisiopatología , Pruebas de Función Cardíaca/efectos de los fármacos , Humanos , Isoleucina/genética , Masculino , Contracción Miocárdica/efectos de los fármacos , Contracción Miocárdica/fisiología , Receptores Adrenérgicos beta 2/metabolismo , Receptores Adrenérgicos beta 2/fisiología , Treonina/genéticaRESUMEN
To control and minimize wear of metal-on-metal hip joints it is essential to understand the mechanisms of debris generation. In vivo, mainly nanosize globular and needle-shaped particles are found. These can neither stem from the action of abrasion nor from tribochemical reactions. In this study the acting wear mechanisms have been first identified on the surface by means of scanning electron microscopy (SEM). Afterwards, the microstructures of the subsurface regions of explants have been investigated using a transmission electron microscope (TEM). Observation of the subsurface gave additional insight about the microstructural changes of cobalt-base alloys subjected to wear. At some distance from the surface, a network of stacking faults and hexagonal epsilon-martensite was found strengthening the bulk material. This microstructure changed into a nanocrystalline type moving closer towards the surface. A comparison of in vivo debris size and grain size of the surface suggests that the globular wear particles result from torn off nanocrystals, while the needle shaped particles are generated by fractured epsilon-martensite. Identified cracks, propagating through the nanocrystalline layer, further support these findings. Thus, it is suggested that the dominating mechanism of particle generation for metal-on-metal joints is surface fatigue within a nanocrystalline surface layer.
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Artroplastia de Reemplazo de Cadera , Metales/química , Adolescente , Adulto , Aleaciones/química , Niño , Preescolar , Análisis de Falla de Equipo , Humanos , Lactante , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Nanoestructuras/química , Estrés MecánicoRESUMEN
1 Cyclic AMP formation has consistently been reported to be desensitized in various tissues including heart of animal models of end-stage renal failure (ESRF). In contrast, reports on desensitization of cAMP formation in ESRF patients remain contradictory. Whether this discrepancy results from a difference between human ESRF and its animal models or from the use of circulating blood cells in the human and various solid tissues in the animal studies, remains unclear. Therefore, we performed three studies with heart and platelets of ESRF patients undergoing haemodialysis or continuous ambulatory peritoneal dialysis and age- and gender-matched controls with normal renal function (n = 11-13 each). 2 In platelets from haemodialysis patients adenylyl cyclase activity in response to receptor-dependent and -independent agonists was reduced by approximately 30%, and this could be explained by an alteration at the level of adenylyl cyclase itself. However, no such desensitization was seen in platelets from peritoneal dialysis patients. 3 In hearts from ESRF patients undergoing haemodialysis, beta-adrenoceptor density and subtype distribution, cAMP formation in response to the beta-adrenoceptor agonist isoprenaline or various receptor-independent stimuli, were very similar to those in control patients but activity of G-protein-coupled receptor kinase was increased by approximately 20%. 4 We conclude that conflicting reports on the desensitization of cAMP formation between ESRF patients and ESRF animal models are not explained by the use of solid tissues in animal studies vs. circulating blood cells in patient studies. Rather desensitization of cAMP formation seems to be a less consistent feature of human ESRF than of its animal models.
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AMP Cíclico/metabolismo , Fallo Renal Crónico/metabolismo , Adenilil Ciclasas/sangre , Apéndice Atrial/efectos de los fármacos , Apéndice Atrial/metabolismo , Apéndice Atrial/cirugía , Plaquetas/química , Plaquetas/metabolismo , Enfermedad Crónica , Quimioterapia/métodos , Femenino , Humanos , Imidazoles/farmacología , Fallo Renal Crónico/fisiopatología , Fallo Renal Crónico/terapia , Masculino , Persona de Mediana Edad , Diálisis Peritoneal Ambulatoria Continua/métodos , Receptores Adrenérgicos beta 1/efectos de los fármacos , Receptores Adrenérgicos beta 1/fisiología , Receptores Acoplados a Proteínas G/metabolismo , Diálisis Renal/métodosRESUMEN
We studied the ontogenetic development of renal alpha 1-, alpha 2-, and beta-adrenergic receptors and their coupling to inositol phosphate and cyclic AMP formation in spontaneously hypertensive and normotensive Wistar-Kyoto rats. alpha 1-, alpha 2-, and beta-Adrenergic receptor number was significantly increased in hypertensive compared with normotensive rats, but the increase did not precede blood pressure elevation. Despite increased alpha 1-adrenergic receptors, basal and norepinephrine-stimulated inositol phosphate formation remained unchanged in all age groups. Rat kidney contains alpha 1A- and alpha 1B-adrenergic receptors coupling to inositol phosphate formation by different mechanisms, but the relative contribution of alpha 1A- and alpha 1B-adrenergic receptors to norepinephrine-stimulated inositol phosphate formation was similar in normotensive and hypertensive rats. Despite increased beta-adrenergic receptors, basal, isoproterenol-, and forskolin-stimulated cyclic AMP accumulation was similar in normotensive and hypertensive rats. We conclude that the number but not the functional responsiveness of renal adrenergic receptors increases in spontaneously hypertensive rats. Thus, the additional receptors are unlikely to contribute to the pathophysiology of elevated blood pressure in this model.
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Riñón/metabolismo , Receptores Adrenérgicos alfa/fisiología , Receptores Adrenérgicos beta/fisiología , Transducción de Señal , Sistema Nervioso Simpático/crecimiento & desarrollo , Animales , AMP Cíclico/metabolismo , Femenino , Fosfatos de Inositol/metabolismo , Masculino , Norepinefrina/farmacología , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Sistema Nervioso Simpático/fisiologíaRESUMEN
beta 1-adrenoceptors play a pivotal role in regulating contractility and heart rate in the human heart. Recently, a polymorphism of the beta 1-adrenoceptor has been detected: at amino acid position 389 either Gly or Arg has been found with the Gly389 exhibiting reduced responsiveness upon agonist-induced stimulation in vitro. In order to find out whether the Gly389 polymorphism exhibits blunted responsiveness also in vivo we studied, in healthy volunteers, the effects of exercise on heart rate and heart rate-corrected duration of electromechanical systole (QS2c as a measure of inotropism) which, in humans, is mediated by beta 1-adrenoceptors stimulation. Twenty-four healthy volunteers (12 female, 12 male) homozygous for the Gly389 or Arg389 exercised on a bicycle in supine position (25, 50, 75 and 100 W for 5 min each), and heart rate and QS2c were assessed; in addition, plasma renin activity (PRA) was determined which is also regulated by beta 1-adrenoceptors in humans. Exercise caused work-load dependent increases in heart rate and PRA, and shortening of QS2c; however, these changes were not significantly different between the Gly389 and Arg389 polymorphism. Thus, these three beta 1-adrenoceptor responses did not differ between volunteers with the Arg389 versus the Gly389 polymorphism. Intragroup analysis, however, revealed that exercise induced increase in heart rate and shortening of QS2c were higher in female than in male volunteers. In conclusion, our data do not support the idea that the reduced responsiveness of Gly389 against agonist-induced stimulation observed in vitro is of major functional importance in vivo.
Asunto(s)
Frecuencia Cardíaca/fisiología , Polimorfismo Genético , Receptores Adrenérgicos beta 1/fisiología , Adulto , Arginina/química , Cartilla de ADN/química , Epinefrina/sangre , Ejercicio Físico/fisiología , Femenino , Glicina/química , Humanos , Masculino , Norepinefrina/sangre , Reacción en Cadena de la Polimerasa , Renina/sangreRESUMEN
1. To elucidate a possible role of species differences in the classification of alpha 1-adrenoceptor subtypes, we have characterized the alpha 1-adrenoceptors in guinea-pig spleen, kidney and cerebral cortex and in bovine cerebral cortex using concentration-dependent alkylation by chloroethylclonidine and competitive binding with 5-methlurapidil, methoxamine, (+)-niguldipine, noradrenaline, oxymetazoline, phentolamine, SDZ NVI-085, tamsulosin and (+)-tamsulosin. Rat liver alpha 1B-adrenoceptors were studied for comparison. Chloroethylclonidine-sensitivity and (+)-niguldipine affinity were also compared at cloned rat and bovine alpha 1a-adrenoceptors. 2. Chloroethylclonidine concentration-dependently inactivated alpha 1-adrenoceptors in all five tissues. While chloroethylclonidine inactivated almost all alpha 1-adrenoceptors in rat liver and guinea-pig kidney and brain, 20-30% of alpha 1-adrenoceptors in guinea-pig spleen and bovine brain were resistant to alkylation by 10 microM chloroethylclonidine. With regard to concentration-dependency guinea-pig kidney and brain were approximately 10 fold less sensitive than guinea-pig spleen or rat liver. 3. In rat liver, all drugs tested competed for [3H]-prazosin binding with steep and monophasic curves. Drug affinities were relatively low and resembled most closely those of cloned rat alpha 1b-adrenoceptors. 4. In guinea-pig spleen, all drugs tested competed for [3H]-prazosin binding with steep and monophasic curves. Drug affinities were relatively low and resembled most closely those of cloned rat alpha 1b-adrenoceptors. 5. In guinea-pig kidney most drugs tested competed for [3H]-prazosin binding with steep and monophasic curves and had relatively low drug affinities close to those of cloned rat alpha 1b- and alpha 1d-adrenoceptors. However, noradrenaline and tamsulosin had consistently biphasic competition curves recognizing 36-39% high and 61-64% low affinity sites. 6. In guinea-pig cerebral cortex, all drugs tested competed for [3H]-prazosin binding with shallow and biphasic curves. While most drugs recognized approximately 25% high affinity sites, tamsulosin and noradrenaline recognized approximately 50% high affinity sites. Drug affinities at the high and low affinity sites except those for tamsulosin and noradrenaline resembled those at cloned alpha 1a- and alpha 1b-adrenoceptors, respectively. 7. In bovine cerebral cortex all drugs tested except for noradrenaline competed for [3H]-prazosin binding with shallow and biphasic curves. All drugs recognized approximately 70% high affinity sites. Drug affinities at the high and low affinity sites resembled those at cloned alpha 1a- and alpha 1b-adrenoceptors, respectively. Noradrenaline competition curves in bovine cerebral cortex were steep and monophasic. 8. When cloned rat and bovine alpha 1a-adrenoceptors transiently expressed in COS cells were studied in a direct side-by-side comparison, both species homologues had similar chloroethylclonidine-sensitivity and (+)-niguldipine affinity. 9. We conclude that properties of bovine alpha 1A- and alpha 1B-adrenoceptors are very similar to those of other species such as rat. alpha 1-Adrenoceptor subtypes in guinea-pigs resemble alpha 1A- and alpha 1B-adrenoceptors in other species but chloroethylclonidine sensitivity and competition binding profiles of noradrenaline and tamsulosin are not compatible with previously established alpha 1-adrenoceptor subtype classification.
Asunto(s)
Receptores Adrenérgicos alfa 1/clasificación , Agonistas alfa-Adrenérgicos/farmacología , Antagonistas Adrenérgicos alfa/farmacología , Animales , Unión Competitiva , Bovinos , Clonidina/análogos & derivados , Clonidina/metabolismo , Cobayas , Técnicas In Vitro , Masculino , Prazosina/metabolismo , Ensayo de Unión Radioligante , Ratas , Ratas Wistar , Receptores Adrenérgicos alfa 1/efectos de los fármacos , Receptores Adrenérgicos alfa 1/metabolismo , Especificidad de la Especie , TritioRESUMEN
Beta2-adrenergic receptors (beta2-AR) contribute to cardiovascular regulation by influencing several functions and previous studies suggest that a decreased function of the beta2-AR may be involved in essential hypertension. Beta2-AR are polymorphic and certain polymorphisms of these receptors are of functional importance. We focus here on the Arg16-->Gly16 beta2-AR polymorphism, which shows enhanced agonist-promoted downregulation of the receptor and which, in two recent studies, yielded opposite results in terms of association with essential hypertension: an increased frequency of the Gly16 variant in African-Caribbean hypertensives and of the Arg16 variant in offspring of Norwegian white hypertensive parents. In the current study, we genotyped 243 subjects, including both African-American and white individuals, for codon 16 polymorphism and assessed blood pressure and cardiovascular function using impedance cardiography and pressor sensitivity to phenylephrine. We found similar patterns of cardiovascular function and expression of hypertension with the two genotypes of codon 16. There was no statistically significant difference in the overall allelic distribution of the two genotypes: among African-Americans, 51% of the hypertensives and 50% of the normotensives carried the Arg16 allele, whereas among the white subjects 40% of the hypertensives and 47% of the normotensives were carriers of the Arg16 allele. Although we observed a statistically significant increase in the Arg16/Gly16 heterozygotes in the African-American population, the Gly16 allele was not significantly increased in the African-Americans compared to whites. These findings indicate that the codon 16 polymorphisms are not associated with hypertension in a mixed American study population nor do they appear to substantially impact on a variety of hemodynamic variables.
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Población Negra/genética , Sistema Cardiovascular/fisiopatología , Codón/genética , Hipertensión/genética , Polimorfismo Genético/genética , Receptores Adrenérgicos beta/genética , Población Blanca/genética , Adulto , Presión Sanguínea , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Hipertensión/etnología , Hipertensión/fisiopatología , Masculino , FenotipoRESUMEN
We have studied the role of alpha 1A- and alpha 1B-adrenoceptors in noradrenaline- and methoxamine-stimulated inositol phosphate accumulation in rat renal cortical slices. [3H]Prazosin binding studies with and without inactivation of alpha 1B-adrenoceptors by chloroethylclonidine treatment suggested that noradrenaline lacks relevant selectivity for alpha 1-adrenoceptor subtypes. Both agonists stimulated [3H]inositol phosphate accumulation with similar maximal effects. The alpha 1A-selective antagonists 5-methyl-urapidil and (+)-niguldipine inhibited inositol phosphate formation by both agonists with shallow biphasic curves but the high affinity component was only 15%-31% and 38%-41%, respectively. The irreversible alpha 1B-selective antagonist chloroethylclonidine inhibited inositol phosphate generation by both agonists by 54%-57%. In contrast to our previous data in rat cerebral cortical slices, we conclude that in rat renal cortex both alpha 1A- and alpha 1B-adrenoceptors are involved in noradrenaline- and methoxamine-stimulated inositol phosphate generation.
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Fosfatos de Inositol/biosíntesis , Corteza Renal/metabolismo , Receptores Adrenérgicos alfa/fisiología , Antagonistas Adrenérgicos alfa/farmacología , Animales , Dihidropiridinas/farmacología , Corteza Renal/ultraestructura , Cinética , Masculino , Metoxamina/farmacología , Norepinefrina/farmacología , Piperazinas/farmacología , Ratas , Ratas Wistar , Especificidad por Sustrato , TritioRESUMEN
We have used the alpha1D-adrenoceptor selective antagonist, BMY 7378, the alpha1D-selective agonists, adrenaline and phenylephrine, the alpha1A-selective antagonists, (+)-niguldipine, SB 216469 and WB4101, and the non-subtype-selective alpha1-adrenoceptor antagonist, nemonapride, to investigate the presence of alpha1D-adrenoceptors in rat tissues at the protein level. Radioligand binding studies using [3H]prazosin as the radioligand were performed in three tissues containing alpha1D-adrenoceptor mRNA, spleen, cerebral cortex and kidney, and in comparison in one tissue not containing alpha1D-adrenoceptor mRNA, liver. Cerebral cortex and kidney were also studied upon alpha1B-adrenoceptor inactivation by chloroethylclonidine treatment (10 microM, 30 min, 37 degrees C). Experiments with cloned rat alpha1-adrenoceptor subtypes transiently expressed in COS cells confirmed the known selectivity of the investigated drugs for alpha1-adrenoceptor subtypes or the lack thereof of nemonapride. Accordingly nemonapride had steep and monophasic competition curves in all native and chloroethylclonidine-treated tissues. BMY 7378 also had steep and monophasic competition curves and low affinity in all native tissues. In contrast, adrenaline and phenylephrine (in the presence of 100 microM GTP) had monophasic competition curves of low affinity in liver and spleen but biphasic competition curves in cerebral cortex and kidney. Following chloroethylclonidine treatment competition curves for adrenaline, phenylephrine, (+)-niguldipine, SB 216469 and WB 4101 remained biphasic in cerebral cortex and kidney while those for nemonapride remained monophasic. We conclude that alpha1D-adrenoceptors are not readily detectable at the protein level in a variety of rat tissues where their mRNA is expressed. The biphasic competition curves of some agonists and antagonists in chloroethylclonidine-treated rat tissues do not represent alpha1D-adrenoceptors and are not readily explained by the present alpha1A/alpha1B/alpha1D-adrenoceptor classification.
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Agonistas alfa-Adrenérgicos/farmacología , Antagonistas Adrenérgicos alfa/farmacología , Corteza Cerebral/efectos de los fármacos , Riñón/efectos de los fármacos , Receptores Adrenérgicos alfa 1/metabolismo , Bazo/efectos de los fármacos , Animales , Corteza Cerebral/metabolismo , Relación Dosis-Respuesta a Droga , Epinefrina/farmacología , Riñón/metabolismo , Masculino , Ratas , Ratas Wistar , Receptores Adrenérgicos alfa 1/efectos de los fármacos , Bazo/metabolismo , Distribución TisularRESUMEN
There can be no doubt that beta(1)-, beta(2)- and beta(3)-adrenoceptor genes have genetic polymorphisms. Two single nucleotide polymorphisms have been described for the beta(1)- (Ser49Gly; Gly389Arg), three for the beta(2)- (Arg16Gly; Gln27Glu; Thr164Ile) and one for the beta(3)-adrenoceptor subtype (Trp64Arg) that might be of functional importance. The possibility that changes in expression or properties of the beta-adrenoceptors due to single nucleotide polymorphisms might have phenotypic consequences influencing their cardiovascular or metabolic function or may contribute to the pathophysiology of several disorders like hypertension, congestive heart failure, asthma or obesity is an idea that has attracted much interest during the last 10 years. At present, it appears that these beta-adrenoceptor polymorphisms are very likely not disease-causing genes, but might be risk factors, might modify disease and/or might influence progression of disease. The aim of this review is to provide an overview of the functional consequences of such beta-adrenoceptor polymorphisms in vitro, ex vivo and in vivo.
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Predisposición Genética a la Enfermedad , Polimorfismo de Nucleótido Simple/genética , Receptores Adrenérgicos beta/genética , Animales , Humanos , Receptores Adrenérgicos beta 1/genética , Receptores Adrenérgicos beta 2/genética , Receptores Adrenérgicos beta 3/genéticaRESUMEN
We have compared the coupling mechanisms of rat renal alpha 1A- and alpha 1B-like adrenoceptors to inositol phosphate formation. The experiments were performed in parallel in native renal tissue preparations and in those where alpha 1B-adrenoceptors had been inactivated by treatment with 10 mumol/l chloroethylclonidine for 30 min at 37 degrees C; renal slices were used in most experiments but isolated renal cells were also used in some cases. The Ca2+ chelating agent, EGTA (5 mmol/l), reduced noradrenaline-stimulated inositol phosphate formation in native but enhanced it in chloroethylclonidine-treated renal slices. The inhibitory effect of EGTA was not mimicked by 100 nmol/l nifedipine. Inactivation of 87% of cellular Gi by 16-20 h treatment with 500 ng/ml pertussis toxin did not significantly affect noradrenaline-stimulated inositol phosphate formation in isolated renal cells but abolished the inhibitory effect of chloroethylclonidine. The adenylate cyclase activator, forskolin (20 mumol/l), inhibited noradrenaline-stimulated inositol phosphate formation in native and chloroethylclonidine-treated slices, and the inhibitory effects of chloroethylclonidine treatment and forskolin were additive. We conclude that in rat kidney inositol phosphate formation via alpha 1B-like adrenoceptors may involve the influx of extracellular Ca2+ and a pertussis toxin-sensitive G-protein but is insensitive to inhibition by forskolin. In contrast alpha 1A-like adrenoceptor-mediated inositol phosphate formation does not require the presence of extracellular Ca2+ or of Gi and is sensitive to inhibition by forskolin. In comparison to published data from other model systems we further conclude that the signaling mechanisms of alpha 1-adrenoceptor subtypes may depend on their cellular environment.
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Riñón/metabolismo , Fosfatidilinositoles/metabolismo , Receptores Adrenérgicos alfa 1/metabolismo , Animales , Calcio/metabolismo , Clonidina/farmacología , Ácido Egtácico , Proteínas de Unión al GTP/metabolismo , Norepinefrina/farmacología , RatasRESUMEN
We have used radioligand binding and inositol phosphate accumulation studies to determine the affinity at mixed alpha 1A- and alpha 1B-adrenoceptors (rat cerebral cortex and kidney), alpha 1A-adrenoceptors (rat cerebral cortex and kidney following inactivation of alpha 1B-adrenoceptors by chloroethylclonidine treatment) and alpha 1B-adrenoceptors (rat spleen) for drugs currently under investigation for the treatment of benign prostatic hypertrophy, alfuzosin, naftopidil and (-)- and (+)-tamsulosin. Alfuzosin and naftopidil had similar affinities in all model systems (approximately 10 nM and 130 nM, respectively) and lacked relevant selectivity for alpha 1-adrenoceptor subtypes. Their potency to inhibit noradrenaline-stimulated inositol phosphate formation in cerebral cortex matched their affinities as determined in the binding studies. Tamsulosin had higher affinity at alpha 1A- than at alpha 1B-adrenoceptors, and was slightly more potent than alfuzosin and naftopidil at alpha 1B- and considerably more potent at alpha 1A-adrenoceptors. However, the interaction of the tamsulosin isomers with chloroethylclonidine-insensitive (alpha 1A-like) adrenoceptors was complex. A detailed analysis of the tamsulosin data and those obtained with other drugs, most notably noradrenaline and oxymetazoline, suggested that chloroethylclonidine-insensitive alpha 1-adrenoceptors may be heterogeneous and that this heterogeneity may differ between cerebral cortex and kidney of the rat.
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Alquilantes/farmacología , Clonidina/análogos & derivados , Riñón/metabolismo , Hiperplasia Prostática/tratamiento farmacológico , Receptores Adrenérgicos alfa 1/metabolismo , Animales , Centrifugación por Gradiente de Densidad , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Clonidina/farmacología , Resistencia a Medicamentos , Técnicas In Vitro , Fosfatos de Inositol/metabolismo , Riñón/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Membranas/efectos de los fármacos , Membranas/metabolismo , Ensayo de Unión Radioligante , Ratas , Ratas Wistar , Receptores Adrenérgicos alfa 1/efectos de los fármacosRESUMEN
SDZ NVI-085 has been proposed to be a centrally acting agonist with selectivity for some alpha 1-adrenoceptor subtypes. We have investigated its selectivity and efficacy at alpha 1-adrenoceptor subtypes in rat tissues and at cloned subtypes. SDZ NVI-085 had higher affinity for chloro-ethylclonidine-insensitive (alpha 1A-like) than for -sensitive (alpha 1B) alpha 1-adrenoceptors in rat kidney but not in cerebral cortex. SDZ NVI-085 recognized cloned alpha 1-adrenoceptor subtypes expressed in COS cells with the order of potency bovine alpha 1C > rat alpha 1A/D > rat alpha 1B. Relative to 100 microM noradrenaline, SDZ NVI-085 was only a partial agonist for stimulation of inositol phosphate formation in rat kidney and inhibited noradrenaline-stimulated inositol phosphate formation in native and chloroethylclonidine-treated tissue. We conclude that SDZ NVI-085 discriminates among multiple alpha 1-adrenoceptor subtypes and is a partial agonist at rat renal alpha 1A- and alpha 1B-adrenoceptors.
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Agonistas alfa-Adrenérgicos/farmacología , Compuestos Bicíclicos con Puentes/farmacología , Hidrocarburos Aromáticos con Puentes , Oxazinas/farmacología , Receptores Adrenérgicos alfa/metabolismo , Agonistas alfa-Adrenérgicos/metabolismo , Animales , Compuestos Bicíclicos con Puentes/metabolismo , Bovinos , Línea Celular , Corteza Cerebral/efectos de los fármacos , Corteza Cerebral/metabolismo , Clonidina/análogos & derivados , Clonidina/farmacología , Técnicas In Vitro , Fosfatos de Inositol/metabolismo , Riñón/efectos de los fármacos , Riñón/metabolismo , Norepinefrina/farmacología , Oxazinas/metabolismo , Prazosina/metabolismo , Ratas , Receptores Adrenérgicos alfa/efectos de los fármacos , Bazo/efectos de los fármacos , Bazo/metabolismo , TransfecciónRESUMEN
To evaluate (1) differences in the peritoneal equilibration test (PET) achieved using continuous peritoneal dialysis (CPD) solutions containing different amounts of glucose and (2) intraindividual reproducibility of PETs performed twice within an interval of 8 months on CPD, we investigated 39 PETs in 13 children aged 2.4-19.0 years (median 10.6 years) on stable CPD regimens. The fill volume was 1 L/m2 body surface area. We used a standard CPD solution (Fresenius) with a 2.3% glucose content (groups 2.3a and 2.3b) two times within an interval of 1-8 months. A third test was done between the two with a CPD solution of 1.5% glucose (group 1.5). Equilibration quotients, that is, substrate concentration in dialysis fluid divided by substrate concentration in plasma (D/P), did not show any statistically significant differences between groups 1.5 and 2.3a or between groups 2.3a and 2.3b. A significant difference was seen in the decline of glucose content of dialysate between groups 1.5 and 2.3 but not between groups 2.3a and 2.3b. Ultrafiltration was higher in groups 2.3a and 2.3b compared with group 1.5. Inter- and intraindividual variability between solute transfer was small during follow-up in stable CPD patients. Different glucose contents of 1.5 and 2.3 g/dL dialysis fluid had no measurable influence on PET results of stable CPD patients. For standard PETs, reducing the glucose content of dialysis fluid to isoosmolarity is not necessary.
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Soluciones para Diálisis , Diálisis Peritoneal Ambulatoria Continua , Peritoneo/metabolismo , Adolescente , Adulto , Niño , Preescolar , Creatinina/metabolismo , Femenino , Humanos , Masculino , Concentración Osmolar , Diálisis Peritoneal Ambulatoria Continua/efectos adversos , Peritonitis/etiología , Peritonitis/metabolismo , Fósforo/metabolismo , Reproducibilidad de los Resultados , Urea/metabolismoRESUMEN
BACKGROUND: Ureteropelvic junction obstruction is the most frequent malformation of the upper urinary tract and treatment with conservative or operative management remains controversial. In this study we present the retrospective analysis of 129 children with ureteropelvic junction obstruction who underwent conservative or operative management. MATERIAL AND METHODS: A total of 129 children with ureteropelvic junction obstruction, who were treated in the department of pediatric nephrology at the University of Essen from 1998-2005, were included into the analysis. Clinical charts were reviewed for the parameters urinary tract infections (total number, severity, bacteriology), antibiotics, ultrasound, Tc-99 diuresis renography, and management (conservative or operative). Statistical analysis was performed using the SPSS software (Version 11.0) RESULTS: A total of 89 urinary tract infections was observed in 52 children. The mean width of renal pelvis was 3.04 ± 1.33 cm in the operative group and 1.98 ± 1.2 cm in the conservative group (p=0.001, ANOVA test). Tc-99 diuresis renography was performed in 70 children of which 46 children received primarily conservative management and 24 children were operated. In the conservative group 6 children underwent pyeloplasty later on due to aggravation of renal function. In 59 out of 129 cases diuresis nephrography was not performed due to only mild ectasia. CONCLUSIONS: This study demonstrates that conservative management is safe in children with ureteropelvic junction obstruction with no or only little constricted renal function, if a close-meshed surveillance protocol is followed and parental compliance is given.
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Antibacterianos/uso terapéutico , Infecciones Bacterianas/tratamiento farmacológico , Hidronefrosis/terapia , Obstrucción Ureteral/terapia , Ureterostomía/estadística & datos numéricos , Infecciones Urinarias/tratamiento farmacológico , Infecciones Bacterianas/complicaciones , Infecciones Bacterianas/diagnóstico , Femenino , Humanos , Hidronefrosis/diagnóstico , Hidronefrosis/etiología , Lactante , Masculino , Estudios Retrospectivos , Resultado del Tratamiento , Obstrucción Ureteral/diagnóstico , Obstrucción Ureteral/etiología , Infecciones Urinarias/complicaciones , Infecciones Urinarias/diagnósticoRESUMEN
OBJECTIVE: IGFBP3 immunoreactivity may appear elevated in patients with chronic kidney disease (CKD), in part due to accumulation of low molecular fragments. The importance of these IGFBP3 variants for binding and inactivation of IGF1 and their relevance for the impaired growth of uremic children are unclear. Nevertheless, IGFBP3, measured as total (t-)IGFBP3, is frequently used as a diagnostic parameter in pediatric CKD patients. A new assay for functional (f-)IGFBP3 exclusively detects IGFBP3 capable of IGF binding. The aim of the study was to evaluate the significance of f-IGFBP3 measurements for the assessment of uremic abnormalities of the GH/IGF1 axis. DESIGN: Prospective cross-sectional study. METHODS: t-IGFBP3, f-IGFBP3, and IGF1 were measured in pediatric CKD patients, including patients with CKD stage 3-4 not on dialysis (CKD, n=33), on dialysis treatment (DT, n=26), patients after renal transplantation (RTx, n=89), healthy children (n=29), children with GH deficiency (GHD, n=42), and small for gestational age (SGA) children (SGA, n=34). RESULTS: Mean t-IGFBP3 SDS was elevated in CKD, DT, and RTx children compared with controls and GHD patients (P≤0.0004). Highest values were reached in DT (P<0.0001 vs all groups). In contrast, mean f-IGFBP3 was similar in all groups (P=0.30). CONCLUSIONS: Pediatric CKD patients displayed elevated serum concentrations of t-IGFBP3 but not f-IGFBP3, supporting the hypothesis that IGFBP3 fragments not binding IGF1 accumulate during uremia. f-IGFBP3 is an indicator of IGFBP3 fragmentation and seems to reflect IGF1 binding in CKD better than t-IGFBP3. However, the role of f-IGFBP3 for the diagnosis of disturbances of the GH/IGF hormonal axis appears to be limited.