[Attention deficit/hyperactivity disorder: burden of the disease according to subtypes in recently diagnosed children]. / Trastorno por déficit de atención/hiperactividad: impacto de la enfermedad de acuerdo al subtipo en niños con diagnóstico reciente.

INTRODUCTION: Attention-Deficit/Hyperactivity Disorder (ADHD) is associated with deterioration of several dimensions of quality of life (QoL) and with the development of comorbid psychiatric disorders. The objective of the present study is to evaluate the burden of illness of ADHD subtypes in untreated newly diagnosed children in Spain. METHODS: We recruited 124 children (80 combined, 25 inattentive and 19 hyperactive-impulsive subtype) aged 6-12 years with untreated newly diagnosed Diagnostic and Statistical Manual of Mental Disorders, 4th edition DSM-IV) ADHD. We collected socio-demographic, clinical (Attention-Deficit/Hyperactivity Disorder Rating Scale ADHD-RS], Conner's Parent Rating Scale-Revised: Short Form [CPRS:R-S], Clinical Global Impression-Severity [CGI-S], Kiddie Schedule for Affective Disorders and Schizophrenia for School Aged Children-Present and Lifetime Version [K-SADS-PL], intelligence Quotient [IQ]), Quality of Life (QoL), Child Health Questionnaire-Parent Form 50 CHQ-PF50), academic performance and health care resources utilization data. We investigated the correlations between ADHD symptom severity and QoL, academic performance and time from onset of symptoms to diagnosis. RESULTS: QoL of children with combined-type ADHD was rated as significantly worse in patients with predominance of hyperactivity/impulsivity for most of the domains. Inattentive-type children also had worse ratings than patients with hyperactivity/impulsivity predominance in most of the domains. The ADHD Index of Conner's Parent Rating Scale-Revised: Short Form (CPRS-R:S) was significantly lower in hyperactive/impulsive patients. We found no differences across subtypes in IQ, academic performance and health care resources utilization. Higher ADHD symptom severity was associated to poor QoL. CONCLUSIONS: Combined and inattentive subtypes are associated with greater disorder severity, more comorbid psychiatric disorders, and worse QoL than the subtype with hyperactivity/impulsivity predominance.

Trastorno por déficit de atención/hiperactividad: impacto de la enfermedad de acuerdo al subtipo en niños con diagnóstico reciente / Attention deficit/hyperactivity disorder: burden of the disease according to subtypes in recently diagnosed children

Introducción. El trastorno por déficit de atención/hiperactividad (TDAH) se asocia con un deterioro en varias dimensiones de calidad de vida (QoL) y el desarrollo de trastornos psiquiátricos comórbidos. El objetivo del presente estudio es evaluar el impacto de la enfermedad de los subtipos de TDAH en niños con diagnóstico inicial y sin tratamiento en España. Métodos. Reclutamos a 124 niños (80 de subtipo combinado, 25 inatento y 19 hiperactividad-impulsividad) entre 6 y 12 años de edad con nuevo diagnóstico de TDAH (Manual diagnóstico y estadístico de los trastornos mentales, 4.a ed., DSM-IV) y sin tratar. Recogimos datos sociodemográficos y clínicos (Attention Deficit/Hyperactivity Disorder-Rating Scale [Escala de evaluación para el trastorno por déficit de atención/hiperactividad, ADHD-RS], Conner’s Parent Rating Scale-Revised: Short Form [Escala de evaluación de Conner, versión para los padres revisada: versión breve, CPRS:R-S], Impresión Clínica Global-Severidad [ICG-S], Kiddie Schedule of Affective Disorders and Schizophrenia for School Aged Children-Present and Lifetie Version [Inventario Kiddie para los trastornos afectivos esquizofrenia para niños en edad escolar, versión actual y a lo largo de la vida, K-SADS-PL] y coeficiente intelectual [CI]), de Qualite of Life (Calidad de vida, Qol), Child Health Questionnaire-Parent Form 50 (Cuestionario de Salud Infantil, versión para los padres de 50 ítems, CHQ-PF50), rendimiento académico y utilización de recursos sanitarios. Investigamos las correlaciones entre la gravedad del TDAH y QoL, rendimiento académico y tiempo transcurrido desde el inicio de los síntomas hasta el diagnóstico. Resultados. La QoL de niños con subtipo combinado de TDAH se valoró como significativamente peor que en pacientes con predominio de hiperactividad/impulsividad en la mayoría de los dominios. Los niños con subtipo inatento también puntuaron peor que los de predominio de hiperactividad/impulsividad en varios dominios. La puntuación de ADHD de la CPRS-R:S fue significativamente menor en los pacientes con predominio de hiperactividad/impulsividad. No encontramos diferencias entre los distintos subtipos respecto al CI, rendimiento académico y utilización de recursos sanitarios. Una mayor gravedad de los síntomas del TDAH se asoció con una peor QoL. Conclusiones. Los subtipos combinado e inatento se asociaron con mayor gravedad del trastorno, más trastornos psiquiátricos comorbidos y peor QoL que el subtipo con predominio de hiperactividad/impulsividad (AU)

Introduction. Attention-Deficit/Hyperactivity Disorder (ADHD) is associated with deterioration of several dimensions of quality of life (QoL) and with the development of comorbid psychiatric disorders. The objective of the present study is to evaluate the burden of illness of ADHD subtypes in untreated newly diagnosed children in Spain. Methods. We recruited 124 children (80 combined, 25 inattentive and 19 hyperactive-impulsive subtype) aged6-12 years with untreated newly diagnosed Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) ADHD. We collected socio-demographic, clinical(Attention-Deficit/Hyperactivity Disorder Rating Scale[ADHD-RS], Conner’s Parent Rating Scale-Revised: Short Form [CPRS:R-S], Clinical Global Impression-Severity [CGI-S], Kiddie Schedule for Affective Disorders and Schizophrenia for School Aged Children-Present and Lifetime Version[K-SADS-PL], intelligence Quotient [IQ]), Quality of Life (QoL), Child Health Questionnaire-Parent Form 50(CHQ-PF50), academic performance and health care resources utilization data. We investigated the correlations between ADHD symptom severity and QoL, academic performance and time from onset of symptoms to diagnosis. Results. QoL of children with combined-type ADHD was rated as significantly worse in patients with predominance of hyperactivivity/impulsivity for most of the domains. Inattentive-type children also had worse ratings than patients with hyperactivity/impulsivity predominance in most of the domains. The ADHD Index of Conner’s Parent Rating Scale-Revised: Short Form (CPRS-R:S) was significantly lower in hyperactive/impulsive patients. We found no differences across subtypes in IQ, academic performance and health care resources utilization. Higher ADHD symptom severity was associated to poor QoL. Conclusions. Combined and inattentive subtypes are associated with greater disorder severity, more comorbid psychiatric disorders, and worse QoL than the subtype with hyperactivity/impulsivity predominance (AU)

Robustness test of a headspace solid-phase microextraction method for the determination of chloroanisoles and chlorophenols related to cork taint in wine using experimental design.

A robustness test of a solid-phase microextraction-based method optimised for the simultaneous determination of chloroanisoles and acetyl-chlorophenols implicated in the presence of corky taste in wine has been carried out using a hybrid experimental design. The influence of small changes around the nominal level of four factors (Vs/Vt ratio, extraction temperature, exposure time and sample incubation time) on the measured response were evaluated in order to indicate if the method is robust for the experimental range considered. Moreover, it was also necessary to identify the critical parameters in the validated model in order to keep them under strict control. Experimental design provides an effective approach for robustness testing as a part of the analytical method validation.

Time-course of oxygen free radical production in acinar cells during acute pancreatitis induced by pancreatic duct obstruction.

The time-course of oxygen free radicals (OFR) generation within acinar cells was studied at different stages of acute pancreatitis (AP) induced in rats by duct obstruction (PDO) for 48 h by flow cytometry, using dihydrorhodamine-123 (DHR) as fluorescent dye. Parallel measurements of the most common markers of oxidative stress such as glutathione (GSH) depletion and malondialdehyde (MDA) levels in pancreas were also performed. OFR production significantly increased within acinar cells at early stages of AP, concomitant with a marked depletion in pancreatic GSH. Lipid peroxidation was significantly enhanced 6 h after PDO, suggesting that the antioxidant defence system of the cell is overwhelmed by OFR production. Both MDA and OFR production in acinar cells decreased to normal values at late AP stages, thus allowing the recovery of pancreatic GSH levels 48 h after PDO. Among the two types of acinar cells differentiated by flow cytometry, R1 and R2, it was the R2 population that showed higher values of DHR dye. However, no differences between the two cell types were found regarding the amount of OFR generation. Our results demonstrate that individual acinar cells significantly contribute to produce large amounts of OFR at early stages of AP. The two existing populations of acinar cells displayed similar behaviour regarding oxidative stress over the course of the disease.

Heterogeneous distribution of plasma membrane glycoconjugates in pancreatic acinar cells.

Flow-cytometric studies of lectin binding to individual acinar cells have been carried out in order to analyse the distribution of membrane glycoconjugates in cells from different areas of the pancreas: duodenal lobule (head) and splenic lobule (body and tail). The following fluoresceinated lectins were used: wheat germ agglutinin (WGA), Tetragonolobus purpureus agglutinin (TP) and concanavalin A (Con A), which specifically bind to N-acetyl D-glucosamine and sialic acid, L-fucose and D-mannose, respectively. In both pancreatic areas, two cell populations (R1 and R2) were identified according to the forward scatter (size). On the basis of their glycoconjugate pattern, R1 cells displayed higher density of WGA and TP receptors than R2 cells throughout the pancreas. Although no difference in size was found between the cells from duodenal and splenic lobules, N-acetyl D-glucosamine and/or sialic acid and L-fucose residues were more abundant in plasma membrane cell glycoconjugates from the duodenal lobule. The results provide evidence for biochemical heterogeneity among individual pancreatic cells according to the distribution of plasma membrane glycoconjugates.

Cell-cycle distribution of pancreatic cells from rats with acute pancreatitis induced by bile-pancreatic obstruction.

The aim of this study was to analyze, using electron microscopy, the morphological alterations that progressively appear in the pancreas of rats with acute pancreatitis induced by bile-pancreatic obstruction over 48 h. In addition, in order to ascertain the capability of pancreas regeneration at different stages of pancreatitis, the distribution of pancreatic cells throughout the different phases of the cell cycle was also analyzed by flow cytometry using propidium iodide staining. Interstitial edema, macrophage infiltration, vacuolization, and dilatation of endoplasmic reticulum were observed from 1.5 h after obstruction onward. Interestingly, cell cycle studies showed an increased proportion of S-phase cells at early stages of pancreatitis (1.5 h after obstruction), which leads to a significant increase in cells in G2/M phase 12 h after pancreatic obstruction. Histological studies revealed severe alterations in pancreas of rats with obstruction maintained over 48 h which affects the nuclear structure. Intracellular disorganization, apoptosis, and focal necrosis were observed at this stage. Furthermore, flow-cytometric analysis of cell DNA contents showed a significant decrease in the proportion of S and G2/M cells and a significant increase in G0/G1 cells, suggesting an arrest of almost all cells in quiescent states. These results suggest that rat pancreas cells are able to recover during the first 12 h after pancreatic obstruction. However, the gland would lose its ability to regenerate if the obstruction was maintained for longer periods.

Alterations in the glycoconjugates of pancreatic cell membrane induced by acute pancreatitis.

The alterations that progressively appear in plasma membrane glycoconjugates of rat pancreatic cells at different stages of acute pancreatitis induced by duct obstruction have been analyzed on individual cells by flow cytometry using the fluoresceinated lectins, wheat germ agglutinin (WGA), Tetragonolobus purpureus agglutinin (TP) and Concanavalin A (Con A), which specifically bind to N-acetyl D-glucosamine, L-fucose and D-mannose, respectively. Two populations of pancreatic cells were differentiated according to the forward scatter (size), which showed different density of saccharidic terminals located at external positions in the glycoconjugates of the plasma membrane. A significant increase in WGA and TP binding was found 1.5 h after pancreatic obstruction, which could be due to the fusion of zymogen granules with the plasma membrane as suggested by the basolateral exocytosis observed by electron microscopy at this stage. The most external sugar residues of membrane glycoconjugates are removed 12 h after pancreatic duct obstruction as a consequence of an advanced state of pancreatitis. The hydrolytic process reaches greater depths in the membrane 48 h after obstruction. At this stage a significant decrease in WGA, TP and ConA binding was found in all pancreatic cells, indicating the loss of N-acetyl D-glucosamine and/or sialic acid, L-fucose and even D-mannose which is located in the core of the glycan. The results provide information about the progressive degradation induced by acute pancreatitis in pancreatic cell membrane glycoconjugates.

Enzyme load in pancreatic acinar cells is increased in the early stages of acute pancreatitis induced by duct obstruction in rats.

Trypsinogen and amylase content has been analysed by flow cytometry in individual pancreatic cells from rats with acute pancreatitis induced by pancreatic duct obstruction, from the earliest stages to 48 h after obstruction. Parallel morphological studies of the pancreas by electron microscopy and analysis of various parameters for the diagnosis of pancreatitis will allow research into the possible relationship between intracellular enzyme load and the severity of pancreatitis. Progressive increases in amylase activity in ascites and plasma, the volume of ascites, haematocrit, vacuolization, oedema and macrophage infiltration were observed between 1.5 h and 12 h after duct obstruction. A progressive increase in enzyme content was also observed in individual acinar cells at this stage. Interestingly, the larger increase was for trypsinogen, so that the trypsinogen/amylase ratio was significantly increased in all acinar cells by 12 h after duct obstruction. This represents a risk factor for the development of pancreatitis. Sections of pancreas taken from rats that had duct obstruction for 48 h showed massive dilatation and disorganization of the endoplasmic reticulum, focal apoptosis and necrosis. These severe alterations would affect enzyme synthesis, as reflected by the significant decrease in the intracellular enzyme load observed at this stage. However, not all acinar cells were affected equally by the damage induced by pancreatitis: R(1) cells appeared to be more sensitive than R(2) cells. In conclusion, intracellular accumulation of digestive enzymes occurs at early stages of pancreatitis, and this effect is proportionally greater for trypsinogen, a finding that could explain the degree of severity achieved in the course of pancreatitis.