Maternal body-mass index and cord blood circulating endothelial colony-forming cells.

OBJECTIVE: Endothelial colony-forming cells (ECFCs) are a subset of circulating endothelial progenitor cells that are particularly abundant in umbilical cord blood. We sought to determine whether ECFC abundance in cord blood is associated with maternal body-mass index (BMI) in nonpathologic pregnancies. STUDY DESIGN: We measured the level of ECFCs in the cord blood of neonates (n = 27) born from non-obese healthy mothers with nonpathologic pregnancies and examined whether ECFC abundance correlated with maternal BMI. We also examined the effect of maternal BMI on ECFC phenotype and function using angiogenic and vasculogenic assays. RESULTS: We observed variation in ECFC abundance among subjects and found a positive correlation between prepregnancy maternal BMI and ECFC content (r = 0.51, P = .007), which was independent of other obstetric factors. Despite this variation, ECFC phenotype and functionality were deemed normal and highly similar between subjects with maternal BMI <25 kg/m(2) and BMI between 25-30 kg/m(2), including the ability to form vascular networks in vivo. CONCLUSIONS: This study underlines the need to consider maternal BMI as a potential confounding factor for cord blood levels of ECFCs in future comparative studies between healthy and pathologic pregnancies.

Biomarkers in Lysosomal Storage Diseases.

A biomarker is generally an analyte that indicates the presence and/or extent of a biological process, which is in itself usually directly linked to the clinical manifestations and outcome of a particular disease. The biomarkers in the field of lysosomal storage diseases (LSDs) have particular relevance where spectacular therapeutic initiatives have been achieved, most notably with the introduction of enzyme replacement therapy (ERT). There are two main types of biomarkers. The first group is comprised of those molecules whose accumulation is directly enhanced as a result of defective lysosomal function. These molecules represent the storage of the principal macro-molecular substrate(s) of a specific enzyme or protein, whose function is deficient in the given disease. In the second group of biomarkers, the relationship between the lysosomal defect and the biomarker is indirect. In this group, the biomarker reflects the effects of the primary lysosomal defect on cell, tissue, or organ functions. There is no "gold standard" among biomarkers used to diagnosis and/or monitor LSDs, but there are a number that exist that can be used to reasonably assess and monitor the state of certain organs or functions. A number of biomarkers have been proposed for the analysis of the most important LSDs. In this review, we will summarize the most promising biomarkers in major LSDs and discuss why these are the most promising candidates for screening systems.

[Variability in the clinical presentation of Pompe disease: development following enzyme replacement therapy]. / Variabilidad en la presentación clínica en la enfermedad de Pompe: evolución tras terapia de reemplazo enzimático.

INTRODUCTION: Pompe disease is a generalized progressive disease caused by a deficiency of the lysosome enzyme acid alpha-glucosidase (GAA). We present three cases with different clinical symptomatology and treated with enzyme replacement therapy (ERT) with positive evolution. CASE REPORTS: Case 1: three-month old male, with weakness and rejecting meals; mild hepatomegaly, discrete macroglossia and muscular hypotony; and increased muscular enzymes. Case 2: five-month old male, with delayed motor development, severe neurosensory deafness, and respiratory disorder of difficult evolution; muscular hypotony; and mild increase in creatine kinase. Case 3: 22-year old male, with progressive dyspnea, with history of increased creatine kinase and transaminases, and hypercholesterolemia. He suffered from severe respiratory failure requiring endotraqueal intubation Muscular biopsy showed glycogen storage suggestive of Pompe disease. In the three cases, the EMG showed a characteristic pattern with pseudomyotonic discharges and the deficiency in GAA was confirmed in lymphocytes. One single mutation was observed in one case and two in the other two cases. Every patient received ERT showing a favorable evolution; with disappearance of cardiac disorders in case 1, improvement in motor development in both infants and no longer need for mechanical ventilation in case 3. CONCLUSION: Pompe disease has a wide variability in clinical expression. ERT achieves a good response, especially in infant forms of the disease. The survival of treated patients with these Pompe disease forms will allow knowing further the course of the disease.

TITLE: Variabilidad en la presentacion clinica en la enfermedad de Pompe: evolucion tras terapia de reemplazo enzimatico.Introduccion. La enfermedad de Pompe es un trastorno generalizado progresivo producido por el deficit de la enzima alfa-glucosidasa acida (AGA) de los lisosomas. Se presentan tres casos manifestados de forma muy diferente y tratados con terapia enzimatica sustitutiva (TES), con evolucion favorable. Casos clinicos. Caso 1: varon de 3 meses, con debilidad y rechazo de la alimentacion, hepatomegalia leve, ligera macroglosia e hipotonia, y aumento de las enzimas musculares. Caso 2: varon de 5 meses, con retraso del desarrollo motor, sordera neurosensorial grave, trastornos respiratorios de repeticion de evolucion torpida, hipotonia y leve elevacion de la creatincinasa. Caso 3: varon de 22 años con disnea progresiva, con antecedentes de elevacion de la creatincinasa y las transaminasas, e hipercolesterolemia. Sufrio insuficiencia respiratoria grave que preciso intubacion endotraqueal. La biopsia muscular presento depositos de glucogeno sugestivos de la enfermedad. En los tres casos, el estudio electromiografico dio un patron caracteristico, con descargas pseudomiotonicas, y se confirmo el deficit de AGA en los linfocitos. Se encontro una mutacion en un caso y dos mutaciones en los otros dos. Todos recibieron TES con evolucion favorable: desaparicion de las alteraciones cardiacas en el caso 1, mejoria en los hitos motores en los dos casos infantiles y retirada del respirador en el caso 3. Conclusion. La enfermedad de Pompe tiene una amplia variabilidad en la expresion clinica. La TES consigue una buena respuesta, especialmente en las formas infantiles. La supervivencia a largo plazo de las formas infantiles tratadas permitira conocer mas aspectos del curso de la enfermedad.

[Cathepsin K as a biomarker of bone involvement in type 1 Gaucher disease]. / Catepsina K como biomarcador de afectación ósea en la enfermedad de Gaucher tipo 1.

BACKGROUND AND OBJECTIVE: Gaucher disease is an inherited disorder caused by deficit of acid ß-glucocerebrosidase, responsible for the degradation of glucosylceramide to ceramide and glucose. Although the disorder is primarily hematologic, bone is the second most commonly affected structure. Cathepsin K (CATK) is an enzyme involved in bone remodelling process. It has been proposed that determination of its serum concentrations may provide additional information to other biomarkers. PATIENTS AND METHODS: The study included 20 control subjects and 20 Gaucher type 1 patients from Andalusia and Extremadura regions. We analyzed the biomarkers of bone remodelling: the bone alkaline phosphatase (B-ALP), the N-terminal propeptide of type 1 procollagen (P1NP), the ß carboxyterminal telopeptide of type 1 collagen (CTx) and the CATK through electrochemiluminescence and immunoassay techniques. RESULTS: There is an increase in levels of CATK, CATK/P1NP and CATK/B-ALP ratios in type 1 Gaucher patients compared to the control group. Considering the existence of skeletal manifestations in the patient group, the CATK and CATK/P1NP ratio showed higher levels in patients with bone damage compared to those without it. CONCLUSIONS: Although imaging studies are the gold standard for monitoring bone disease in type 1 Gaucher patients, the utility of CATK should be considered as a possible indicator of bone damage in these patients. Furthermore, this parameter can be used in the monitoring of the treatment of bone pathology.

Evaluation of a multiplex ELISA for autoantibody profiling in patients with autoimmune connective tissue diseases.

The performance of immunoassays for the detection of autoantibodies is of critical importance in the diagnosis and assessment of patients with autoimmune connective tissue diseases (ACTD). Our objective was to compare the features of two multiplexed assays-INNO-LIA ANA and Gennova-PictArray ENA ELISA-for measurement of multiple autoantibodies and their utility as a clinical tool in ACTD diagnosis. The antigens included SS-A/Ro (60 and 52), SSB/La, Sm, Sm/RNP, CENP-B, Jo-1, and Scl-70. Stored sera from 85 ACTD patients and 80 controls consisting of patients with vasculitis, rheumatoid arthritis and infectious diseases, as well as healthy subjects were analyzed jointly with clinical and laboratory data. Agreement between the two methods varied between 58 and 99% (Cohen's kappa: 0.21-0.71) mostly for SSA and SSB. The frequency of specific autoantibodies measured using the two methods was more variable for SSA, SSB, and RNP/Sm. There were a higher number of ambiguous results when using INNO-LIA. The optimized cut-off values of the Gennova-PictArray resulted in over 99% specificities in samples obtained from the control group. Sensitivity patterns were more accurate in Gennova-PictArray than in INNO-LIA, as suggested in previously reported studies. A third method could be applied to determine which of the two methods is more accurate.

Detection of c. -32T>G (IVS1-13T>G) mutation of Pompe disease by real-time PCR in dried blood spot specimen.

BACKGROUND: Pompe disease, or acid maltase deficiency, is a genetic muscle disorder caused by mutations in the gene encoding the acid alpha-glucosidase (GAA) enzyme, which is essential for the degradation of glycogen to glucose in lysosomes. The wide clinical variability is resulted from genetic heterogeneity, and many different mutations of the GAA gene have been reported. Some of these mutations are associated with specific phenotypes, such as the c. -32T>G (IVS1-13T>G) mutation seen in late-onset Pompe disease. METHODS: We used a real-time PCR, after genomic DNA extraction isolated from DBS (dried blood spots) and PCR amplification. RESULTS: Our results successfully detected in controls and patients have been 100% concordant with sequencing results. CONCLUSIONS: This assay combines simple sample processing and rapid analysis and it allows to detect the patients with a milder form and slower progression of this disease with a high reliability.

[Tetra-saccharide glucose as a diagnostic biomarker for Pompe disease: a study with 35 patients]. / Glucosa tetrasacárido como biomarcador diagnóstico de la enfermedad de Pompe: estudio en 35 pacientes.

BACKGROUND AND OBJECTIVES: Pompe disease is a disorder originating from an acid alpha-glycosidase (AAG) enzyme deficiency. This disease produces an accumulation of lysosomal glycogen in different tissues, whereby the skeletal and heart muscles are especially involved. The established diagnosis is achieved through the identification of the AAG deficiency. There are also other secondary diagnostic biomarkers, such as tetra-saccharide glucose (Glc4), which shows high levels in the urine of these patients. In this study it is highlighted the usefulness of Glc4 as a diagnostic biomarker for Pompe disease in its different forms of presentation, using a high-performance liquid chromatography with ultraviolet detection (HPLC/UV) adapted to the study. PATIENTS AND METHODS: A total of 75 individuals have been analyzed: 40 healthy controls and 35 patients diagnosed with Pompe disease. Twenty-four hour samples of urine were collected from all of the patients and their Glc4 levels were determined by means of HPLC/UV. RESULTS: The evaluation of the urinary Glc4 shows a high discrimination ability between healthy/sick individuals. In addition, the results obtained have allowed to establish the most appropriate level of decision or cut-off point for the identification of sick people. CONCLUSIONS: Glc4 urinary levels are found to be high in patients suffering from Pompe disease and even though increased levels are also found in other conditions, the existence of a AAG deficiency together with a compatible clinical symptoms, prove very helpful for a correct diagnosis of this serious disease.

[Left ventricular mass, diastolic function and collagen metabolism biomarkers in essential hypertension]. / Masa ventricular izquierda, función diastólica y marcadores de metabolismo del colágeno en la hipertensión arterial refractaria.

BACKGROUND AND OBJECTIVE: To evaluate the association between circulating biomarkers of collagen metabolism in serum, left ventricular mass index (LVMI) and diastolic dysfunction in patients with resistant hypertension. PATIENTS AND METHODS: Fifty-two patients with resistant hypertension and 24 healthy individuals were included. The following biomarkers of collagen metabolism were analyzed by ELISA: carboxy-terminal propeptide of procollagen type I (PICP) and transforming growth factor beta1 (TGFß1). The biomarker C-terminal telopeptide of collagen type-I (ICTP) was assayed by electrochemiluminescence immunoassay. In the patient's group a record of 24-h blood pressure monitoring was obtained and an echocardiography was performed. Left ventricular mass was measured according to the formula of Devereux and the diastolic function according to the relation of E and A waves and mitral propagation velocity. RESULTS: Hypertensive patients showed higher levels of PICP and lower levels of ICTP than controls: 83.7 (24.7) vs. 55.0 (8.7), P<.0001; and 175.0 (136.4) vs. 323.3 (121.3), P<.0001). Hypertensive patients showed a significant relationship between PICP and LVMI (r=0.631, P<.0001) and between PICP and diastolic dysfunction (r=-0.519, P<.0001). The groups with and without hypertrophy, and with or without diastolic dysfunction, differed in the mentioned peptides but not in BP values. CONCLUSIONS: Our findings suggest that the analyzed markers of synthesis and degradation of collagen may be related to myocardial hypertrophy and diastolic dysfunction independent of blood pressure values.

Influence of IL28B polymorphisms on response to a lower-than-standard dose peg-IFN-α 2a for genotype 3 chronic hepatitis C in HIV-coinfected patients.

BACKGROUND: Data on which to base definitive recommendations on the doses and duration of therapy for genotype 3 HCV/HIV-coinfected patients are scarce. We evaluated the efficacy of a lower peginterferon-α 2a dose and a shorter duration of therapy than the current standard of care in genotype 3 HCV/HIV-coinfected patients. METHODS AND FINDINGS: Pilot, open-label, single arm clinical trial which involved 58 Caucasian HCV/HIV-coinfected patients who received weekly 135 µg peginterferon-α 2a plus ribavirin 400 mg twice daily during 20 weeks after attaining undetectable viremia. The relationships between baseline patient-related variables, including IL28B genotype, plasma HCV-RNA, ribavirin dose/kg, peginterferon-α 2a and ribavirin levels with virological responses were analyzed. Only 4 patients showed lack of response and 5 patients dropped out due to adverse events related to the study medication. Overall, sustained virologic response (SVR) rates were 58.3% by intention-to-treat and 71.4% by per protocol analysis, respectively. Among patients with rapid virologic response (RVR), SVR and relapses rates were 92.6% and 7.4%, respectively. No relationships were observed between viral responses and ribavirin dose/kg, peginterferon-α 2a concentrations, ribavirin levels or rs129679860 genotype. CONCLUSIONS: Weekly 135 µg pegIFN-α 2a could be as effective as the standard 180 µg dose, with a very low incidence of severe adverse events. A 24-week treatment duration appears to be appropriate in patients achieving RVR, but extending treatment up to just 20 weeks beyond negativization of viremia is associated with a high relapse rate in those patients not achieving RVR. There was no influence of IL28B genotype on the virological responses. TRIAL REGISTRATION: ClinicalTrials.gov NCT00553930.

Olive oil polyphenols decrease blood pressure and improve endothelial function in young women with mild hypertension.

BACKGROUND: Olive oil polyphenols have been associated with several cardiovascular health benefits. This study aims to examine the influence of a polyphenol-rich olive oil on blood pressure (BP) and endothelial function in 24 young women with high-normal BP or stage 1 essential hypertension. METHODS: We conducted a double-blind, randomized, crossover dietary-intervention study. After a run-in period of 4 months (baseline values), two diets were used, one with polyphenol-rich olive oil (∼30 mg/day), the other with polyphenol-free olive oil. Each dietary period lasted 2 months with a 4-week washout between diets. Systolic and diastolic BP, serum or plasma biomarkers of endothelial function, oxidative stress, and inflammation, and ischemia-induced hyperemia in the forearm were measured. RESULTS: When compared to baseline values, only the polyphenol-rich olive oil diet led to a significant (P < 0.01) decrease of 7.91 mm Hg in systolic and 6.65 mm Hg of diastolic BP. A similar finding was found for serum asymmetric dimethylarginine (ADMA) (-0.09 ± 0.01 µmol/l, P < 0.01), oxidized low-density lipoprotein (ox-LDL) (-28.2 ± 28.5 µg/l, P < 0.01), and plasma C-reactive protein (CRP) (-1.9 ± 1.3 mg/l, P < 0.001). The polyphenol-rich olive oil diet also elicited an increase in plasma nitrites/nitrates (+4.7 ± 6.6 µmol/l, P < 0.001) and hyperemic area after ischemia (+345 ± 386 perfusion units (PU)/sec, P < 0.001). CONCLUSIONS: We concluded that the consumption of a diet containing polyphenol-rich olive oil can decrease BP and improve endothelial function in young women with high-normal BP or stage 1 essential hypertension.

Abnormal levels of antioxidant defenses in a large sample of patients with hypertensive disorders of pregnancy.

Increased levels of oxidative stress have been demonstrated in Preeclampsia in previous studies, but this finding has not been established in other hypertensive disorders in pregnancy (HDP). We measured different markers of lipid peroxidation and antioxidant defenses by spectrophotometry or enzymoimmunoanalysis in 339 pregnant women: 85 with gestational hypertension (GH), 88 chronic hypertension (CH), 104 Preeclampsia and 62 healthy pregnant control women (PCW). Lower activity of superoxide dismutase and higher levels of catalase were found in GH, CH and preeclampsia compared with PCW (964.4±116.5, 970.0±120.4, 971.2±137.5 and 1063.4±133.7 U g(-1) Hb, P<0.001; and 313.0±71.7, 292.2±45.3, 297.1±47.2, 215.5±26.2 U mg(-1) Hb, P<0.0001; respectively). Regarding the glutathione REDOX cycle, we found the following in GH, CH and preeclampsia compared with PCW: a decrease in its reduced form (2.6±0.6, 2.7±0.8, 2.7±0.9, 3.3±1.3 µmol l(-1), P<0.003), a parallel increase in the oxidized form (185.6±68.9, 194.7±75.0, 184.3±78.3, 85.1±27.5 µmol l(-1), P<0.0001) and an increment in glutathione peroxidase (85.9±22.0, 86.4±20.9, 82.1±23.5 and 77.2±19.7 U g(-1) Hb, P<0.04) and glutathione reductase (6384.3±1261.9, 6724.6±1154.1, 6287.9±1399.9 and 6044.4±1208.4 mU g(-1) Hb, P<0.01, respectively). Nitrites/nitrates were higher in patients with preeclampsia than in PCW (31.50±15.08, 26.80±8.39 µmol l(-1), P<0.002). Although malondialdehyde and oxidized-LDL levels were similar among groups, free fatty acids were increased in every HDP (GH 514.6±194.6, CH 501.3±197.4, preeclampsia 555.2±230.1 µmol l(-1)) compared with PCW (351.4±146.1 µmol l(-1)), P<0.0001. Our results show an oxidation/reduction imbalance with an increase in oxidative stress coupled with a decreased capacity of antioxidant systems, not only in preeclampsia but also in every HDP.

Estudio comparativo de los niveles de procalcitonina en el plasma y las muestras de sangre seca mediante electroquimioluminiscencia (ECLIA) / A comparative study of procalcitonin levels in plasma and dried blood specimens using electrochemiluminescence (ECLIA)

Introducción. Los procesos febriles constituyen uno de los motivos de consulta más frecuentes en las urgencias pediátricas, siendo las infecciones bacterianas una de las principales causas responsables de los mismos. Su diagnóstico es un reto a superar pues representan una importante causa de morbimortalidad. Entre los biomarcadores séricos utilizados, es la procalcitonina la que aparece en el suero con anterioridad a otros biomarcadores inflamatorios agudos tales como el número de leucocitos en circulación periférica y la proteína C reactiva. Material y métodos. Se utilizaron muestras de 135 pacientes pediátricos recogidas mediante punción en el dedo o talón. Las determinaciones se realizaron respectivamente en plasma y sangre seca recogida sobre papel (DBS), en la misma muestra de cada uno de ellos. Resultados. El estudio estadístico comparativo realizado entre los resultados obtenidos en ambos tipos de muestras, nos indica la existencia de una correlación estadísticamente significativa entre muestras analizadas por ambas metodologías. El test U de Mann-Whitney aplicado asimismo, manifiesta la no existencia de diferencias estadísticamente significativas entre los resultados. Conclusiones. Las muestras DBS tienen una gran utilidad para la cuantificación de los niveles de procalcitonina en la población pediátrica donde, con frecuencia, la extracción de sangre venosa presenta dificultades. La obtención de este tipo de muestra por simple punción en el talón o yema del dedo evita este problema, contribuyendo al diagnóstico rápido de los procesos infecciosos. No obstante, podría mejorarse la sensibilidad del método realizando determinaciones seriadas en aquellas muestras con valores iniciales bajos (menores de 0,650 ng/mL) (AU)

Introduction. Fever processes constitute one of the main reasons for pediatric emergency department visits, being bacterial infections one of the principal causes of this process. Its diagnosis represents a challenge to overcome, due to the high morbidity and mortality rates related to these infections. Among analyzed serum biomarkers, procalcitonin shows an earlier appearance in serum than others inflammatory biomarkers such as peripheral blood leukocytes number and C-reactive protein. Materials and methods. Blood samples were collected by finger or heel puncture from 135 pediatric patients. Procalcitonin was measured in plasma and in dried blood specimen (DBS) for each blood sample. Results. The comparative statistical analysis has revealed a significant correlation between the two procalcitonin assays. The Mann-Whitney U test has demonstrated that there was no difference on the procalcitonin results obtained by both techniques. Conclusions. DBS represent a useful tool for quantifying procalcitonin levels in pediatric population, where venous blood collection is usually difficult. The extraction of this type of blood sample, by finger or heel puncture, avoids this problem, contributing to a rapid diagnosis of this kind of infectious processes. However, method sensitivity could be improved by performing serial measurements in samples with low basal levels (lower than 0.650 ng/mL) (AU)

Valores de referencia de la actividad enzimática alfa-glucosidasa ácida linfocitaria / Reference values of intralymphocyte acid alpha-glucosidase activity

Introducción. La enfermedad de Pompe, también denominada déficit de maltasa ácida o glucogenosis tipo ii, es un trastorno metabólico autosómico recesivo caracterizado por un acúmulo anormal de glucógeno lisosomal, causado por la deficiencia de la enzima α-glucosidasa ácida (GAA). Según la edad de inicio y el grado de afectación orgánica, la enfermedad de Pompe se suele subdividir en 2 tipos: neonatal (infantile-onset) y tardío (late-onset). El diagnóstico clínico se confirma mediante la ausencia virtual (forma infantil) o marcada reducción (forma del adulto) de la actividad enzimática GAA en diferentes muestras biológicas. Material y métodos. El objetivo de este trabajo consiste en establecer los valores de referencia para la actividad enzimática GAA intralinfocitaria que constituye el método gold standard para el diagnóstico de la enfermedad. Resultados. Los resultados obtenidos en una población de sujetos control difieren de los publicados en otros trabajos, lo que refleja la importancia de establecer valores de referencia dentro de cada laboratorio (AU)

Introduction. Pompe disease also called acid maltase deficiency or glycogenosis type ii is an autosomal recessive metabolic disorder characterized by an abnormal accumulation of lysosomal glycogen caused by deficiency of the enzyme acid α-glucosidase (GAA). According to the age of onset and the extent of organ involvement, Pompe disease is usually divided into 2 subtypes: Neonatal (infantile-onset) and adult (late-onset). The clinical diagnosis is confirmed by the virtual absence (infantile form) or markedly reduced (adult form) GAA enzyme activity in different biological samples. Material and methods. The aim of this work is to establish reference values for intralymphocyte alpha-glucosidase acid activity, which is the 'gold standard' method for the diagnosis of Pompe disease. Results. The results obtained in a population of control subjects are different from those published in other studies. This reflects the importance of establishing reference values within each laboratory (AU)

Catepsina K como biomarcador de afectación ósea en la enfermedad de Gaucher tipo 1 / Cathepsin K as a biomarker of bone involvement in type 1 Gaucher disease

Fundamento y objetivo: La enfermedad de Gaucher es un trastorno hereditario, que se origina como consecuencia del déficit de la actividad β-glucocerebrosidasa ácida, responsable de la degradación de glucosilceramida hasta ceramida y glucosa. Aunque el trastorno de base es fundamentalmente hematológico, el hueso es la segunda estructura más frecuentemente afectada. La catepsina K (CATK) es una enzima implicada en el proceso de remodelado óseo, habiéndose propuesto que la determinación de sus concentraciones séricas podría aportar información complementaria a la de otros biomarcadores. Pacientes y métodos: Se realizó un estudio en 20 controles sanos y 20 pacientes con enfermedad de Gaucher tipo 1, de las comunidades autónomas de Andalucía y Extremadura. Se determinaron como biomarcadores de remodelado óseo la bone alkaline phosphatase (B-ALP, «fosfatasa alcalina ósea»), el amino-terminal propeptide of procollagen type 1 (P1NP, «propéptido aminoterminal del procolágeno 1»), la β-Cross Laps, carboxy-terminal telopeptide of collagen type 1 (CTx, «fracción β del colágeno tipo 1») y CATK por técnicas de electroquimioluminiscencia y enzimoinmunoanálisis. Resultados: Existe un incremento en los niveles de CATK y las ratios CATK/P1NP y CATK/B-ALP en los pacientes con Gaucher tipo 1 respecto a la media obtenida en el grupo control. Por otro lado, considerando la existencia o no de manifestaciones óseas en el grupo de pacientes, la CATK y la ratio CATK/P1NP muestran niveles medios superiores en aquellos pacientes con daño óseo respecto a los que no lo presentan. Conclusiones: Aunque los estudios radiológicos constituyen el gold-standard para el seguimiento de enfermedad ósea en pacientes con enfermedad de Gaucher tipo 1, debe considerarse la utilidad de la CATK como posible indicador de daño óseo en estos pacientes. Asimismo, este parámetro puede utilizarse en la monitorización del tratamiento de la enfermedad ósea (AU)

Background and objective: Gaucher disease is an inherited disorder caused by deficit of acid β-glucocerebrosidase, responsible for the degradation of glucosylceramide to ceramide and glucose. Although the disorder is primarily hematologic, bone is the second most commonly affected structure. Cathepsin K (CATK) is an enzyme involved in bone remodelling process. It has been proposed that determination of its serum concentrations may provide additional information to other biomarkers. Patients and methods: The study included 20 control subjects and 20 Gaucher type 1 patients from Andalusia and Extremadura regions. We analyzed the biomarkers of bone remodelling: the bone alkaline phosphatase (B-ALP), the N-terminal propeptide of type 1 procollagen (P1NP), the β carboxyterminal telopeptide of type 1 collagen (CTx) and the CATK through electrochemiluminescence and immunoassay techniques. Results: There is an increase in levels of CATK, CATK/P1NP and CATK/B-ALP ratios in type 1 Gaucher patients compared to the control group. Considering the existence of skeletal manifestations in the patient group, the CATK and CATK/P1NP ratio showed higher levels in patients with bone damage compared to those without it. Conclusions: Although imaging studies are the gold standard for monitoring bone disease in type 1 Gaucher patients, the utility of CATK should be considered as a possible indicator of bone damage in these patients. Furthermore, this parameter can be used in the monitoring of the treatment of bone pathology (AU)

Variabilidad en la presentación clínica en la enfermedad de Pompe: evolución tras terapia de reemplazo enzimático / Variability in the clinical presentation of Pompe disease: development following enzyme replacement therapy

Introducción. La enfermedad de Pompe es un trastorno generalizado progresivo producido por el déficit de la enzima alfaglucosidasa ácida (AGA) de los lisosomas. Se presentan tres casos manifestados de forma muy diferente y tratados con terapia enzimática sustitutiva (TES), con evolución favorable. Casos clínicos. Caso 1: varón de 3 meses, con debilidad y rechazo de la alimentación, hepatomegalia leve, ligera macroglosia e hipotonía, y aumento de las enzimas musculares. Caso 2: varón de 5 meses, con retraso del desarrollo motor, sordera neurosensorial grave, trastornos respiratorios de repetición de evolución tórpida, hipotonía y leve elevación de la creatincinasa. Caso 3: varón de 22 años con disnea progresiva, con antecedentes de elevación de la creatincinasa y las transaminasas, e hipercolesterolemia. Sufrió insuficiencia respiratoria grave que precisó intubación endotraqueal. La biopsia muscular presentó depósitos de glucógeno sugestivos de la enfermedad. En los tres casos, el estudio electromiográfico dio un patrón característico, con descargas pseudomiotónicas, y se confirmó el déficit de AGA en los linfocitos. Se encontró una mutación en un caso y dos mutaciones en los otros dos. Todos recibieron TES con evolución favorable: desaparición de las alteraciones cardíacas en el caso 1, mejoría en los hitos motores en los dos casos infantiles y retirada del respirador en el caso 3. Conclusión. La enfermedad de Pompe tiene una amplia variabilidad en la expresión clínica. La TES consigue una buena respuesta, especialmente en las formas infantiles. La supervivencia a largo plazo de las formas infantiles tratadas permitirá conocer más aspectos del curso de la enfermedad (AU)

Introduction. Pompe disease is a generalized progressive disease caused by a deficiency of the lysosome enzyme acid alphaglucosidase (GAA). We present three cases with different clinical symptomatology and treated with enzyme replacement therapy (ERT) with positive evolution. Case reports. Case 1: three-month old male, with weakness and rejecting meals; mild hepatomegaly, discrete macroglossia and muscular hypotony; and increased muscular enzymes. Case 2: five-month old male, with delayed motor development, severe neurosensory deafness, and respiratory disorder of difficult evolution; muscular hypotony; and mild increase in creatine kinase. Case 3: 22-year old male, with progressive dyspnea, with history of increased creatine kinase and transaminases, and hypercholesterolemia. He suffered from severe respiratory failure requiring endotraqueal intubation Muscular biopsy showed glycogen storage suggestive of Pompe disease. In the three cases, the EMG showed a characteristic pattern with pseudomyotonic discharges and the deficiency in GAA was confirmed in lymphocytes. One single mutation was observed in one case and two in the other two cases. Every patient received ERT showing a favorable evolution; with disappearance of cardiac disorders in case 1, improvement in motor development in both infants and no longer need for mechanical ventilation in case 3. Conclusion. Pompe disease has a wide variability in clinical expression. ERT achieves a good response, especially in infant forms of the disease. The survival of treated patients with these Pompe disease forms will allow knowing further the course of the disease (AU)

Glucosa tetrasacárido como biomarcador diagnóstico de la enfermedad de Pompe: estudio en 35 pacientes / Tetra-saccharide glucose as a diagnostic biomarker for Pompe disease: A study with 35 patients

Fundamento y objetivos: La enfermedad de Pompe es un trastorno originado por la deficiencia de la enzima alfa glucosidasa ácida (GAA). En esta afección se produce un acúmulo de glucógeno lisosomal en diferentes tejidos, estando especialmente implicados los músculos esquelético y cardíaco. El diagnóstico de confirmación se realiza mediante identificación del déficit de GAA. Existen, además, otros biomarcadores diagnósticos secundarios, como la glucosa tetrasacárido (Glc4), que se muestra elevada en orina de estos pacientes. Así, con este trabajo queremos poner de manifiesto la utilidad de la Glc4 como biomarcador diagnóstico para la enfermedad de Pompe en sus diferentes formas de presentación, utilizando un método de high-performance liquid chromatography (HPLC, «cromatografía líquida de alta resolución») con detección ultravioleta (UV) adaptado para nuestro estudio. Pacientes y métodos: Hemos analizado un total de 75 individuos: 40 controles sanos y 35 pacientes diagnosticados de enfermedad de Pompe. Se han recogido muestras de orina de 24 h de todos ellos y se han determinado sus niveles de Glc4 mediante HPLC/UV. Resultados: La evaluación de la Glc4 urinaria muestra una gran capacidad de discriminación entre individuos sanos/enfermos. Además, los resultados obtenidos nos han permitido establecer el nivel de decisión o punto de corte más apropiado para la identificación de los enfermos. Coclusiones: Los niveles de Glc4 urinarios se encuentran elevados en los pacientes con enfermedad de Pompe, y aunque se encuentran incrementados en otras dolencias, la existencia de un déficit de GAA, junto a una clínica compatible, proporcionan una alta sensibilidad para el diagnóstico de esta grave enfermedad (AU)

Background and objectives: Pompe disease is a disorder originating from an acid alpha-glycosidase (AAG) enzyme deficiency. This disease produces an accumulation of lysosomal glycogen in different tissues, whereby the skeletal and heart muscles are especially involved. The established diagnosis is achieved through the identification of the AAG deficiency. There are also other secondary diagnostic biomarkers, such as tetra-saccharide glucose (Glc4), which shows high levels in the urine of these patients. In this study it is highlighted the usefulness of Glc4 as a diagnostic biomarker for Pompe disease in its different forms of presentation, using a high-performance liquid chromatography with ultraviolet detection (HPLC/UV) adapted to the study. Patients and methods: A total of 75 individuals have been analyzed: 40 healthy controls and 35 patients diagnosed with Pompe disease. Twenty-four hour samples of urine were collected from all of the patients and their Glc4 levels were determined by means of PLC/UV. Results: The evaluation of the urinary Glc4 shows a high discrimination ability between healthy/sick individuals. In addition, the results obtained have allowed to establish the most appropriate level of decision or cut-off point for the identification of sick people. Conclusions: Glc4 urinary levels are found to be high in patients suffering from Pompe disease and even though increased levels are also found in other conditions, the existence of a AAG deficiency together with a compatible clinical symptoms, prove very helpful for a correct diagnosis of this serious disease (AU)

Masa ventricular izquierda, función diastólica y marcadores de metabolismo del colágeno en la hipertensión arterial refractaria / Left ventricular mass, diastolic function and collagen metabolism biomarkers in essential hypertension

Fundamento y objetivo: Evaluar la asociación entre biomarcadores del metabolismo del colágeno, índice de masa ventricular izquierda (IMVI) y función diastólica en pacientes con hipertensión arterial (HTA) refractaria. Pacientes y método:Se estudiaron 52 pacientes diagnosticados de HTA refractaria y se compararon con 24 individuos sanos. Se midió en suero el propéptido C-terminal de la molécula de procolágeno tipo I (PICP) y el factor de crecimiento transformante beta 1 (TGFβ1) por métodos de enzimoinmunoanálisis, y el telopéptido C-terminal del colágeno tipo I (ICTP) por inmunoensayo electroquimioluminiscente. A los pacientes se les practicó una ecocardiografía donde se calculó el IMVI por la fórmula de Devereux y se valoró la función diastólica a partir de la relación entre las ondas E y A (E/A) y la velocidad de propagación mitral. También se les practicó un registro de monitorización de la presión arterial (PA) de 24h. Resultados:Los hipertensos mostraron valores (media ±DE) superiores de PICP e inferiores de ICPT que los controles: 83,7 (24,7) frente a 55,0 (8,7), p<0,0001, y 175,0 (136,4) frente a 323,3 (121,3), p<0,0001. En los hipertensos existió una relación significativa entre el PICP y el IMVI (r=0,631, p<0,0001) y disfunción diastólica (r=-0,519, p<0,0001). Los grupos con y sin hipertrofia, y con o sin función diastólica, diferían en los citados péptidos pero no en las cifras de PA. Conclusiones: Nuestros hallazgos sugieren que diferentes marcadores de la síntesis y de la degradación del colágeno pueden relacionarse con la presencia de hipertrofia miocárdica o disfunción diastólica con independencia de las cifras de PA (AU)

To evaluate the association between circulating biomarkers of collagen metabolism in serum, left ventricular mass index (LVMI) and diastolic dysfunction in patients with resistant hypertension.Patients and methods: Fifty-two patients with resistant hypertension and 24 healthy individuals were included. The following biomarkers of collagen metabolism were analyzed by ELISA: carboxy-terminal propeptide of procollagen type I (PICP) and transforming growth factor beta1 (TGFβ1). The biomarker C-terminal telopeptide of collagen type-I (ICTP) was assayed by electrochemiluminescence immunoassay. In the patient's group a record of 24-h blood pressure monitoring was obtained and an echocardiography was performed. Left ventricular mass was measured according to the formula of Devereux and the diastolic function according to the relation of E and A waves and mitral propagation velocity. Results:Hypertensive patients showed higher levels of PICP and lower levels of ICTP than controls: 83.7 (24.7) vs. 55.0 (8.7), P<.0001; and 175.0 (136.4) vs. 323.3 (121.3), P<.0001). Hypertensive patients showed a significant relationship between PICP and LVMI (r=0.631, P<.0001) and between PICP and diastolic dysfunction (r=-0.519, P<.0001). The groups with and without hypertrophy, and with or without diastolic dysfunction, differed in the mentioned peptides but not in BP values. Conclusions:Our findings suggest that the analyzed markers of synthesis and degradation of collagen may be related to myocardial hypertrophy and diastolic dysfunction independent of blood pressure values (AU)

TGF-beta 1 y su relación con la hipertensión arterial esencial / Transforming growth factor-Beta1 in essential hypertension

Introducción. Se ha sugerido que los niveles plasmáticos elevados del factor de crecimiento transformante beta 1 (TGF-beta 1) juegan un papel clave para el desarrollo de las enfermedades que cursan con fibrosis. Objetivos. Este estudio observacional, transversal, retrospectivo de casos controles busca evaluar si existe asociación entre los niveles plasmáticos de TGF-beta 1 como factor causal en la patogénesis de la hipertensión arterial (HT-A) esencial y la lesión de órganos diana, en pacientes con HT-A esencial refractaria comparado con un grupo de sujetos sanos y comprobar si existe una correlación entre los niveles de TGF-beta 1 y los de PICP (propéptido C-terminal de la molécula de procolágeno tipo I), de degradación MMP-1 (metaloproteinasa de la matriz tipo 1) y degradación del colágeno ICTP (telopéptido C-terminal de la molécula de colágeno tipo I) en el grupo de sujetos hipertensos. Pacientes y métodos. Se estudiaron 52 pacientes diagnosticados de HT-A con edad media de 53 años y se comparó con grupo control de 24 voluntarios sanos con edad media de 45 años. TGF-beta 1 fue medido por método ELISA previa activación de la muestra. Resultados. Las concentraciones de TGF-beta 1 no fueron mayores en el grupo de hipertensos. La media±desviación estándar fue de 40±13 pg/mL, mientras que en el grupo control fue de 50±23 pg/mL. La t de Student no estableció diferencias significativas. Conclusiones. no se han encontrado niveles elevados de TGF-beta 1 en pacientes con hipertensión esencial en contra de lo evidenciado por otros autores. La exclusión en este estudio de pacientes que presentaban insuficiencia renal moderada o severa, asumimos que pudiera ser la causa de no encontrar elevados los niveles plasmáticos de TGF-beta (AU)

Introduction. It has been suggested that elevated serum transforming growth factor beta1 (TGF-beta 1) levels plays a key role to develope of diseases associated with fibrosis. Objective. This observational, transversal, retrospective case control study was designed to evaluate the association between TGF-beta 1 as causal factor for the hypertension arterial pathogenesis and damage on target organ. This study was designed to evaluate the association between circulating biomarkers of collagen metabolism and fibrosis in serum to compare TGF-beta1 levels obtained in a group of essential hypertension patients with a group of normotensive healthy subjects. Patients and methods. 46 essential hypertension patients, mean age: 53 years versus a control group of 20 healthy volunteers, mean age: 45 years. TGFâ1 was quantitated by a commercial ELISA technique, samples were previously activated. Results. TGF-beta 1 concentrations were not higher in essential hypertension patients, mean serum concentration (40±13ng/mL) than in normal group, mean serum concentration (50±23ng/mL). No significant differences were showed between two groups using t- student test. Conclusion. We have not found elevated TGF-beta 1 concentrations in essential hypertension patients in contrast to what has been shown by other authors. Exclusion of patients with mild or severe renal insufficiency should be considered to be a cause for not obtaining elevated TGF-beta 1 concentrations(AU)

La actividad ß-galactosidasa como índice de control de calidad de la muestra de sangre seca recogida sobre papel / Beta-galactosidase activity as an index of quality control of dried blood sample collected on paper

Las enfermedades de depósito lisosomal son errores congénitos del metabolismo originados por la deficiencia hereditaria de hidrolasas lisosomales, causando un acúmulo progresivo de moléculas complejas que no pueden degradarse. El diagnóstico definitivo de estas patologías consiste en la determinación de la actividad enzimática específica en leucocitos, fibroblastos u otros tipos celulares, además del diagnóstico molecular. Estos métodos son complejos y presentan incomodidades para los pacientes. En los últimos años, la determinación de la actividad enzimática en muestras de sangre seca recogida sobre papel ha facilitado enormemente tanto la toma de muestras, su envío a los centros de referencia, así como la manipulación de las mismas. En este trabajo, presentamos la determinación de la actividad enzimática β-galactosidasa en muestras de sangre seca recogida sobre papel, como un parámetro de gran utilidad que nos asegura la estabilidad de la muestra remitida a los laboratorios para el diagnóstico de cualquier tipo de enfermedad lisosomal (AU)

The lysosomal storage diseases are inborn errors of the metabolism originated by the hereditary deficiency of lysosomal hydrolases, which cause a progressive accumulation of complex molecules that cannot be degraded. Besides the molecular diagnosis, the definitive diagnosis of these pathologies consists of the determination of specific enzymatic activities in leukocytes, fibroblasts or other types of cells. These methods are complex and may be inconvenient for the patients. In the last few years, the determination of enzymatic activity in dried blood specimen (DBS) has facilitated sampling, their shipment to the reference laboratories, as well as their manipulation. In this work, we present the determination of the enzymatic β-galactosidase activity in DBS as a very useful parameter to ensure the stability of the sample sent to the laboratories for the diagnosis of any lysosomal disease (AU)