Four Years' Experience in the Diagnosis of Very Long-Chain Acyl-CoA Dehydrogenase Deficiency in Infants Detected in Three Spanish Newborn Screening Centers.

Identification of very long-chain acyl-CoA dehydrogenase deficiency is possible in the expanded newborn screening (NBS) due to the increase in tetradecenoylcarnitine (C14:1) and in the C14:1/C2, C14:1/C16, C14:1/C12:1 ratios detected in dried blood spots. Nevertheless, different confirmatory tests must be performed to confirm the final diagnosis. We have revised the NBS results and the results of the confirmatory tests (plasma acylcarnitine profiles, molecular findings, and lymphocytes VLCAD activity) for 36 cases detected in three Spanish NBS centers during 4 years, correlating these with the clinical outcome and treatment. Our aim was to distinguish unambiguously true cases from disease carriers in order to obtain useful diagnostic information for clinicians that can be applied in the follow-up of neonates identified by NBS.Increases in C14:1 and of the different ratios, the presence of two pathogenic mutations, and deficient enzyme activity in lymphocytes (<12% of the intra-assay control) identified 12 true-positive cases. These cases were given nutritional therapy and all of them are asymptomatic, except one. Seventeen individuals were considered disease carriers based on the mild increase in plasma C14:1, in conjunction with the presence of only one mutation and/or intermediate residual activity (18-57%). In addition, seven cases were classified as false positives, with normal biochemical parameters and no mutations in the exonic region of ACADVL. All these carriers and the false positive cases remained asymptomatic. The combined evaluation of the acylcarnitine profiles, genetic results, and residual enzyme activities have proven useful to definitively classify individuals with suspected VLCAD deficiency into true-positive cases and carriers, and to decide which cases need treatment.

[Ischemia-reperfusion syndrome and role of preservation graft technique after laparoscopic versus open nephrectomy in a experimental model of living donor kidney transplant]. / Evaluación del síndrome isquemia-reperfusión y papel de la preservación del injerto en la extracción renal laparoscópica versus abierta en un modelo experimental porcino de donante vivo.

INTRODUCTION: Delayed graft function alter living donor transplantation is a subject of debate. Delayed graft function can be partially explained by renal ischemia-reperfusion injury, when severe is associated with decreased graft survival. In this experimental living donor model study, we analyze the hemodynamic, histological and biochemical effects of laparoscopic nephrectomy. We also, analyze the effect of a pulsatile machine perfusion for kidney preservation during cold ischemia time. MATERIAL AND METHODS: Twenty large-white pigs (average weight 40-45 kgrs) were divided in 4 experimental groups: Group A: Laparoscopic nephrectomy+ immediate graft perfusion in pulsatile vacuum pump+autotransplant Group B: Laparoscopic nephrectomy+ immediate graft perfusion by gravity+autotransplant Group C: Open nephrectomy+immediate graft perfusion in pulsatile vacuum pump+autotransplant Group D: Open nephrectomy+ immediate graft perfusion by gravity+autotransplant Both laparoscopic and open nephrectomy were completed transperitoneally according to standardized technique. Hypothermic perfusion was done in a system designed in our lab. RESULTS: We observed a decreased renal artery flow in kidneys procured laparoscopically compared to open nephrectomy. We found an artery flow recovery during the first 60 minutes after revascularization. Renal machine perfusion during cold ischemia time seems to have no beneficial effect, but shows a deleterious effect on hemodynamic event for renal transplantation. Lower plasma nitric oxide level is observed in kidneys obtained by laparoscopy compared with open surgical technique. And finally, we also found higher histological damage in proximal tubular and endothelial cell, in kidneys obtained by laparoscopy compared with open surgery. CONCLUSIONS: In our experience: Laparoscopic nephrectomy versus open nephrectomy produces, in a model of living donor transplant, a lower value or renal blood flow and a higher value of renal vascular resistanse. These hemodynamic findings tend to normalize by 60 min after the reperfusion. A lower blood concentration of nitric oxide after the transplant was detected in laparoscopic group Vs open surgery group.

Evaluación del síndrome isquemia-reperfusión y papel de la preservación del injerto en la extracción renal laparoscópica versus abierta en un modelo experimental porcino de donante vivo / Ischemia-reperfusion syndrome and role of preservation graft technique after laparoscopic versus open nephrectomy in a experimental model of living donor kidney transplant

Introducción: La disfunción inicial del injerto renal extraído por laparoscopia en un donante vivo es un aspecto que, aún hoy, crea ciertas controversias. El síndrome de isquemia-reperfusión explicaría parcialmente esta disfunción inicial del injerto trasplantado que, según su intensidad, puede llegar a condicionar una menor supervivencia del riñón. Analizamos en un modelo experimental de donante vivo la repercusión hemodinámica, bioquímica e histológica de la extracción laparoscópica del injerto. Como objetivo secundario evaluamos el papel de la aplicación de una técnica de preservación (perfusión en bomba) durante los minutos de isquemia fría. Material y metodos: Utilizamos 20 cerdos tipo large-white (peso 40-45 kgrs), que fueron divididos en 4 grupos: Grupo A: Extracción laparoscópica + perfusión inmediata del injerto tras la extracción mediante bomba pulsátil + Autotrasplante Grupo B: Extracción laparoscópica + perfusión por gravedad del injerto + Autotrasplante Grupo C: Extracción por cirugía abierta + perfusión inmediata del injerto tras la extracción mediante bomba pulsátil + Autotrasplante Grupo D: Extracción por cirugía abierta + perfusión por gravedad del injerto + Autotrasplante Las extracciones laparoscópica y por cirugía abierta se realizaron por vía transperitoneal con técnica estandarizada y sistemática. La perfusión hipotérmica en bomba se llevó a cabo mediante un sistema pulsátil diseñado por nuestro grupo. Resultados. Comprobamos que el flujo arterial del riñón trasplantado que previamente fue extraído por laparoscopia, es menor que los extraídos por cirugía abierta, y existe una tendencia a recuperarse tras la primera hora post-reperfusión. La aplicación de un periodo corto de preservación del injerto durante la isquemia fría no mejora el patrón hemodinámico del injerto trasplantado, incluso resulta perjudicial. Por otro lado, la concentración de óxido nítrico en sangre venosa del injerto extraído por laparoscopia fue menor que la de los obtenidos mediante cirugía abierta. Finalmente, el estudio histológico mostró peor conservación de los elementos túbulo-glomerular y endotelial en los órganos extraídos por laparoscopia. Conclusiones: En nuestra experiencia: - La extracción laparoscópica renal en un modelo de donante vivo frente a la extracción abierta, determina un menor flujo renal y mayor resistencia vascular. - Esta alteración hemodinámica de los riñones extraídos por laparoscopia tiene tendencia a corregirse a los 60 minutos tras la reperfusión. - Existe una menor concentración sanguínea de NO en los injertos trasplantados que fueron extraídos por laparoscopia en comparación con el grupo extraídos por cirugía abierta

Introduction: Delayed graft function alter living donor transplantation is a subject of debate. Delayed graft function can be partially explained by renal ischemia-reperfusion injury, when severe is associated with decreased graft survival. In this experimental living donor model study, we analyze the hemodynamic, histological and biochemical effects of laparoscopic nephrectomy. We also, analyze the effect of a pulsatile machine perfusion for kidney preservation during cold ischemia time. Material and methods: Twenty large-white pigs (average weight 40-45 kgrs) were divided in 4 experimental groups: Group A: Laparoscopic nephrectomy+ immediate graft perfusion in pulsatile vacuum pump+autotransplant Group B: Laparoscopic nephrectomy+ immediate graft perfusion by gravity+autotransplant Group C: Open nephrectomy+immediate graft perfusion in pulsatile vacuum pump+autotransplant Group D: Open nephrectomy+ immediate graft perfusion by gravity+autotransplant Both laparoscopic and open nephrectomy were completed transperitoneally according to standardized technique. Hypothermic perfusion was done in a system designed in our lab. RESULTS. We observed a decreased renal artery flow in kidneys procured laparoscopically compared to open nephrectomy. We found an artery flow recorvery during the first 60 minutes after revascularization. Renal machine perfusion during cold ischemia time seems to have no beneficial effect, but shows a deleterious effect on hemodynamic event for renal transplantation. Lower plasma nitric oxide level is observed in kidneys obtained by laparoscopy compared with open surgical technique. And finally, we also found higher histological damage in proximal tubular and endothelial cell, in kidneys obtained by laparoscopy compared with open surgery. Conclusions: In our experience: - Laparoscopic nephrectomy versus open nephrectomy produces, in a model of living donor transplant, a lower value or renal blood flow and a higher value of renal vascular resístanse. - These hemodynamic findings tend to normalize by 60 min after the reperfusion. - A lower blood concentration of nitric oxide after the transplant was detected in laparoscopic group Vs open surgery group

Sildenafil improves immediate posttransplant parameters in warm-ischemic kidney transplants: experimental study.

OBJECTIVE: To evaluate in an experimental model the effects of the PDE5 inhibitor sildenafil on kidney grafts autotransplanted after a period of 45 minutes of warm ischemia and 60 minutes of hypothermic pump perfusion. METHODS: Nine laboratory large-white pigs were divided into two groups. Group A (n = 4): oral dose of 100 mg sildenafil was administered 1 hour before the surgery. Group B (n = 5): no sildenafil given. Right single nephrectomy was completed after a 45-minute period of warm ischemia by complete vascular clamping. Before the autotransplant, all kidneys were submitted to a 60-minute period of hypothermic pulsatile perfusion. Renal flow, arterial pressure, and renal vascular resistance were recorded in real time for 60 minutes after autotransplant. Nitric oxide levels were determined in blood samples of the renal vein at predefined intervals. Optical and electronic microscopy was performed on all organs at the end of the procedure. RESULTS: Renal vascular flow was significantly higher and renal vascular resistance significantly lower in the sildenafil group compared with the non-sildenafil group. No significant differences were observed in systemic arterial pressure values between both groups. Nitric oxide levels were significantly higher for all periods in the sildenafil group. No differences were observed in histological studies. CONCLUSION: Our experimental work suggested a positive effect of sildenafil on the immediate posttransplant outcome of warm-ischemic kidneys without systemic secondary effects.

Cribado neonatal de las enfermedades congénitas / Neonatal screening for congenital diseases

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Déficit de 21-hidroxilasa: aspectos actuales / 21-hydroxylase deficiency: current aspects

La hiperplasia suprarrenal congénita (HSC) es una de las alteraciones autosómicas recesivas más frecuentes. Caracterizada por un defecto enzimático en la síntesis de cortisol, la causa es, en el 95% de los casos, la deficiencia de la enzima 21-hidroxilasa (21-OH). La 17-OH progesterona, precursor del cortisol, presenta valores elevados, marcadores del diagnóstico. Esta enfermedad presenta diferentes formas clínicas; las clásicas o graves comienzan desde el período neonatal, con síntomas debidos al exceso de andrógenos suprarrenales como virilización y ambigüedad de los genitales externos de las niñas afectadas. En más del 70% de los casos se asocia pérdida salina (deficiencia de aldosterona), potencialmente letal en varones que no se diagnostican precozmente. Resumimos las diferentes formas de presentación de la deficiencia de 21-OH, y describimos el diagnóstico y el tratamiento con gluco y mineralcorticoides, con especial énfasis en la importancia de utilizar dosis de estrés de hidrocortisona, cuando es necesario. Los avances quirúrgicos actuales ofrecen una corrección funcional de las pacientes afectadas. Los programas de detección precoz evitan la asignación incorrecta de sexo en la recién nacida y pueden salvar la vida de los varones con formas graves y pérdida salina. Comentamos el diagnóstico genético-molecular del CYP21A2 (cromosoma 6p 21.3) y las características en la población española. Revisamos las directrices futuras para el estudio y el tratamiento de esta enfermedad, incluyendo diversos tratamientos como la flutamida, la hormona de crecimiento, los antagonistas de las gonadotropinas o la relación con el síndrome de ovario poliquístico. El diagnóstico y el tratamiento prenatales del feto femenino afectado son posibles, y los resultados son alentadores. Comentamos, también, el abordaje hacia la transición y la edad adulta, y la relevancia del control de la mujer con HAC durante la gestación

Congenital adrenal hyperplasia (CAH) is one of the most frequent autosomal recessive disorders. It is characterized by a deficiency of an enzyme involved in cortisol synthesis and in 95% of patients the cause is 21-hydroxylase deficiency. A diagnostic marker is elevated levels of 17-hydroxyprogesterone, a precursor of cortisol. CAH has several clinical forms, and classical or severe forms manifest in the neonatal period with symptoms due to excess adrenal androgen production such as virilization and ambiguity of the external genitalia in affected girls. In more than 70% of patients, there is associated salt wasting (aldosterone deficiency), which can be fatal in males without an early diagnosis. We summarize the various forms of presentation of 21-hydroxylase deficiency and describe diagnosis and treatment with gluco- and mineralocorticoids, with special emphasis on the importance of using stress doses of hydrocortisone when necessary. Current surgical advances provide functional correction in affected patients. Screening programs avoid incorrect sex assignment in the newborn and can save the lives of males with severe forms and salt wasting. We discuss the genetic-molecular diagnosis of CYP21A2 (chromosome 6p 21.3) and its characteristics in the Spanish population. We review future recommendations for the study and management of this disease, including several treatments such as flutamide, growth hormone, and gonadotrophin antagonists, as well as the association with polycystic ovary syndrome. Prenatal diagnosis and treatment in affected female fetuses are feasible and the results are encouraging. We also discuss the management of the transition to adulthood and the importance of follow-up of women with CAH during pregnancy

Grupo de consensosobre toxoplasmosis / Consensus group on toxoplasmosis

El objetivo de este grupo de trabajo ha sido conseguir un consenso sobre qué debe o no debe hacerse frente a la toxoplasmosis congénita. Según los conocimientos actuales, que a continuación se resumen, se ha podido llegar a las siguientes conclusiones: -- Las medidas higiénicas y los hábitos culinarios que forman parte de la prevención primaria de la toxoplasmosis están recomendados en todo embarazo, son de aplicación muy simple y su beneficio es indudable.-- Los programas de prevención secundaria de la toxoplasmosis congénita (cribado sistemático en la población gestante y prevención, diagnóstico y tratamiento de la infección fetal) no cumplen, en el momento actual, los criterios fundamentales para que se realicen como medida de salud pública en España. Se desconoce el impacto poblacional de este tipo de programas.-- Debe considerarse la oportunidad de implantar programas de prevención terciaria mediante cribado neonatal de la toxoplasmosis. (AU)

Isolated kidney controlled perfusion with true physiological pulsatile waveform.

A computer controlled perfusion system has been developed to study the behavior of perfused kidneys in several conditions. The system is designed to perform kidney perfusions at constant pressure and low temperature (about 4 degrees C). We compared 2 types of perfusion pumps, a classical roller pump widely used in hemodialysis circuits and a vacuum powered tubular pump with active valves developed by our group and able to produce a flow pattern very similar to the pulse wave in the human circulatory system. In this preliminary study, we show the hydrodynamics obtained with both pumps in isolated hypothermic kidneys perfused with this system. The different flow patterns with both pumps seem to determine differences in the preservation conditions of the kidney.

[Prenatal molecular genetic diagnosis and treatment of congenital adrenal hyperplasia due to 21-hydroxylase deficiency]. / Diagnóstico genético molecular y tratamiento prenatal de la hiperplasia adrenal congénita por déficit de la enzima 21-hidroxilasa.

Congenital adrenal hyperplasia due to 21-hydroxylase deficiency (21-OH), in its severe forms, produces virilization of the external genitalia of the affected female fetus. Early treatment with dexamethasone of the pregnant mother at risk of a fetus with 21-OH deficiency avoids the masculinization of the affected female fetus. We present a pregnant mother, where the prenatal diagnosis was established by DNA analysis of a chorionic villous sample obtained in the 9th week of gestation. Molecular analysis showed the female fetus to be affected of 21-OH deficiency. Maternal treatment with dexamethasone started on the 6th week of gestation has prevented the virilization of the affected baby. No significant side effects have been encountered. Prenatal diagnosis and treatment for 21-OH deficiency is effective and safe, as is described in the literature. This is the first case in Spain where both prenatal molecular diagnosis and treatment for 21-OH deficiency have been reported.