RESUMEN
BACKGROUND: There are only a few series in the literature on acute disseminated encephalomyelitis (ADEM) in children. OBJECTIVES AND METHODS: the aims of this study were to perform (i) a prospective clinical/imaging study (1992-2009) on ADEM in children consecutively referred to our institution in Catania, Italy, and (ii) to undertake a systematic review and meta-analysis of published ADEM pediatric cohorts (>10 cases). RESULTS: We identified 17 patients with ADEM (incidence <10 years of age=1.1 per 100 000 person-years). 15 previously published cohorts were compared with our cohort: (i) systematically reviewed (750 cases); and (ii) meta-analyzed (492/750 cases). The 17 patients had the following characteristics: (a) male-to-female ratio, 1.4 (vs. 1.2-1.3 in previous cohorts); (b) mean age at presentation, 3.6 years (vs. 7.1 years in previous cohorts); (c) specific preceding triggering factor, 88% (vs. 69-79% in previous cohorts); (d) the most common initial signs were ataxia, seizures, headache, and thalamic syndrome; (e) brain imaging revealed >3 lesions in 100% (vs. 92% in previous cohorts); (f) the outcome was good in 94% (vs. 70-75% in previous cohorts); and (g) 12% relapsed once (vs. 18% in previous cohorts). CONCLUSIONS: ADEM is generally a benign condition that mosly affects boys more than girls and rarely recurs.
Asunto(s)
Encéfalo/patología , Encefalomielitis Aguda Diseminada/diagnóstico , Corticoesteroides/uso terapéutico , Edad de Inicio , Encéfalo/fisiopatología , Niño , Electroencefalografía , Encefalomielitis Aguda Diseminada/tratamiento farmacológico , Encefalomielitis Aguda Diseminada/fisiopatología , Femenino , Glucocorticoides/uso terapéutico , Humanos , Masculino , Pronóstico , Estudios Prospectivos , Resultado del TratamientoRESUMEN
A time-delayed fluorescence immunoassay was developed for the determination of serum levels of methylglyoxal (MG)-derived hydroimidazolone using a monoclonal antiserum raised against Nalpha-acetyl-Ndelta-(5-hydro-5-methyl)-4-imidazolone, Europium-labeled anti-mouse IgG antiserum as indicator, and MG modified bovine serum albumin (BSA) as standard. Serum levels of hydroimidazolone were measured in 45 patients with type 2 diabetes aged 59.4 +/- 6.1 (mean +/- SD) years and with duration of diabetes of 7.3 +/- 3.1 years, and in 19 nondiabetic controls aged 56.3 +/- 4.3 years. The serum levels of hydroimidazolone were significantly higher in patients compared to controls: median, 3.0 (5-95 percentile, 1.6 to 5.4) U/mg protein versus 1.9 (1.2 to 2.8) U/mg protein (P =.0005). Significant positive correlations were observed between the serum levels of hydroimidazolone and serum levels of advanced glycation end products (AGEs), measured with a polyclonal anti-AGE antibody: r = 0.59 for patients (P <.0001), and r = 0.65 for controls (P =.002). Similarly, significant correlations were also found between serum levels of hydroimidazolone and N(epsilon)-(carboxymethyl)-lysine (CML): r = 0.36 in patients and r = 0.55 for controls (both P =.02). Serum hydroimidazolone levels did not correlate with fasting plasma glucose or hemoglobin A(1c) (HbA(1c)) levels. The observed differences between patients with diabetes and nondiabetic controls seem to be comparable to differences measured for other AGE compounds.
Asunto(s)
Diabetes Mellitus Tipo 2/sangre , Imidazoles/sangre , Lisina/análogos & derivados , Lisina/sangre , Piruvaldehído/metabolismo , Humanos , Inmunoensayo/normas , Sensibilidad y EspecificidadRESUMEN
Recently, all the structural features of non-proliferative diabetic retinopathy have been demonstrated in mice fed 30% galactose for 21-26 months. To determine whether changes in retinal matrix protein mRNA levels occur early in the course of murine diabetes we used a competitive RT-PCR method to quantitate retinal mRNA levels in an inbred mouse strain (FVB) commonly used for transgenic studies. Retinal mRNA was prepared from STZ-diabetic and non-diabetic FVB mice at 4, 8, 12 and 16 weeks and cDNA encoding basement membrane components was quantitated using MIMIC constructs that compete for the same primer pairs. alpha1 (IV) collagen, the beta1 and gamma1 chains of laminin, fibronectin, and vitronectin mRNAs were quantitated. For alpha1 (IV) collagen, statistically significant diabetes-induced increases were apparent by 8 weeks (3.11 +/- 0.20 vs. 1.29 +/- 0.19 x 10(6) molecules/mg total RNA, p < 0.005). Similarly, diabetes-induced increases were observed by 8 weeks for the beta1 chain of laminin (4.54 +/- 0.22 vs. 1.85 +/- 0.43 x 10(5) molecules/mg total RNA, p < 0.005), the gamma1 chain of laminin (7. 33 +/- 0.29 vs. 4.84 +/- 0.76 x 10(4)/microg total RNA, p < 0.05), and for fibronectin (2.22 +/- 0.21 vs. 1.35 +/- 0.15 x 10(6) molecules/mg total RNA, p < 0.05). The magnitude of change was greatest for alpha1 (IV) collagen (2.4-fold) and beta1 laminin (2. 5-fold) at 8 weeks, and least for fibronectin (1.6-fold). A smaller diabetes-induced increase in vitro nectin mRNA was also observed, but it failed to reach statistical significance at 12 and 16 weeks. These data provide the basis for assessing the effects of genetic manipulation on diabetic retinopathy in transgenic mouse models.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Proteínas de la Matriz Extracelular/genética , ARN Mensajero/metabolismo , Retina/metabolismo , Animales , Colágeno/genética , Fibronectinas/genética , Laminina/genética , Masculino , Ratones , Ratones Endogámicos , Ratones Transgénicos , Valores de Referencia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Vitronectina/genéticaRESUMEN
Diabetic hyperglycaemia causes a variety of pathological changes in small vessels, arteries and peripheral nerves. Vascular endothelial cells are an important target of hyperglycaemic damage, but the mechanisms underlying this damage are not fully understood. Three seemingly independent biochemical pathways are involved in the pathogenesis: glucose-induced activation of protein kinase C isoforms; increased formation of glucose-derived advanced glycation end-products; and increased glucose flux through the aldose reductase pathway. The relevance of each of these pathways is supported by animal studies in which pathway-specific inhibitors prevent various hyperglycaemia-induced abnormalities. Hyperglycaemia increases the production of reactive oxygen species inside cultured bovine aortic endothelial cells. Here we show that this increase in reactive oxygen species is prevented by an inhibitor of electron transport chain complex II, by an uncoupler of oxidative phosphorylation, by uncoupling protein-1 and by manganese superoxide dismutase. Normalizing levels of mitochondrial reactive oxygen species with each of these agents prevents glucose-induced activation of protein kinase C, formation of advanced glycation end-products, sorbitol accumulation and NFkappaB activation.
Asunto(s)
Hiperglucemia/etiología , Mitocondrias/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Animales , Ácido Aspártico/metabolismo , Glucemia/metabolismo , Carbonil Cianuro m-Clorofenil Hidrazona/farmacología , Proteínas Portadoras/farmacología , Bovinos , Transporte de Electrón , Complejo II de Transporte de Electrones , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Endotelio Vascular/patología , Activación Enzimática , Productos Finales de Glicación Avanzada/metabolismo , Hiperglucemia/metabolismo , Hiperglucemia/patología , Canales Iónicos , Malatos/metabolismo , Proteínas de la Membrana/farmacología , Proteínas Mitocondriales , Complejos Multienzimáticos/metabolismo , FN-kappa B/metabolismo , Oxidorreductasas/metabolismo , Proteína Quinasa C/metabolismo , Rotenona/farmacología , Sorbitol/metabolismo , Succinato Deshidrogenasa/metabolismo , Superóxido Dismutasa/metabolismo , Superóxido Dismutasa/farmacología , Tenoiltrifluoroacetona/análogos & derivados , Tenoiltrifluoroacetona/farmacología , Desacopladores/farmacología , Proteína Desacopladora 1RESUMEN
Idiopathic left ventricular tachycardia is a rare arrhythmia whose electrophysiological basis is not yet well-defined. We report a case of idiopathic left ventricular tachycardia caused by a reentrant circuit limited exclusively to the two fascicles of the left bundle branch.
Asunto(s)
Fascículo Atrioventricular/fisiopatología , Taquicardia Paroxística/diagnóstico , Taquicardia Ventricular/diagnóstico , Adulto , Estimulación Cardíaca Artificial , Errores Diagnósticos , Electrocardiografía , Humanos , Masculino , Taquicardia Paroxística/fisiopatología , Taquicardia Supraventricular/diagnóstico , Taquicardia Ventricular/fisiopatologíaRESUMEN
Aminoguanidine (AG) treatment, like nerve growth factor (NGF) treatment, prevents diabetes-induced apoptosis of retinal Müller cells in the rat eye, but the mechanism involved is unknown. In this study, the effects of preincubation with AG on oxidant-induced apoptosis, oxidant-induced intracellular reactive oxygen species (ROS) production, and lipid peroxidation were determined in rat retinal Müller cells and compared with the effects of NGF, a protein that protects neuronal cells from oxidative stress. The effect of AG on rabbit vitreous lipid peroxide levels was also determined. After exposure to increasing concentrations of H2O2, there was a corresponding increase in the percentage of apoptotic Müller cells. Preincubation with AG for 48 h completely inhibited oxidant-induced apoptosis in response to 10 micromol/l H2O2 (+AG 0 vs. 10 micromol/l, NS), and reduced the percentage of apoptotic cells in response to 50 micromol/l H2O2 by 50% (+AG vs. -AG, P < 0.01). Longer preincubation did not increase the antiapoptotic effect of AG. The effect of AG was dose-dependent. Similar results were obtained after preincubation with NGF. Both AG and NGF preincubation prevented the twofold increase in oxidant-induced lipid peroxides. The fivefold increase in oxidant-induced ROS production was decreased 100% by NGF, but only 61% by AG preincubation. The twofold increase in vitreous lipid peroxide level in diabetic rabbits was completely prevented by AG treatment. AG reduced H2O2-induced benzoate hydroxylation in a dose-dependent manner. Intracellular glutathione content was unchanged. These data demonstrate that AG can act as an antioxidant in vivo, quenching hydroxyl radicals and lipid peroxidation in cells and tissues and preventing oxidant-induced apoptosis.
Asunto(s)
Apoptosis/efectos de los fármacos , Guanidinas/farmacología , Peroxidación de Lípido/efectos de los fármacos , Oxidantes/farmacología , Especies Reactivas de Oxígeno/metabolismo , Retina/metabolismo , Animales , Diabetes Mellitus Experimental/metabolismo , Inhibidores Enzimáticos/farmacología , Peróxido de Hidrógeno/farmacología , Radical Hidroxilo/metabolismo , Factores de Crecimiento Nervioso/farmacología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Conejos , Ratas , Cuerpo Vítreo/metabolismoRESUMEN
Methylglyoxal (MG), a dicarbonyl compound produced by the fragmentation of triose phosphates, forms advanced glycation endproducts (AGEs) in vitro. Glyoxalase-I catalyzes the conversion of MG to S-D-lactoylglutathione, which in turn is converted to D-lactate by glyoxalase-II. To evaluate directly the effect of glyoxalase-I activity on intracellular AGE formation, GM7373 endothelial cells that stably express human glyoxalase-I were generated. Glyoxalase-I activity in these cells was increased 28-fold compared to neo-transfected control cells (21.80+/-0.1 vs. 0. 76+/-0.02 micromol/min/mg protein, n = 3, P < 0.001). In neo-transfected cells, 30 mM glucose incubation increased MG and D-lactate concentration approximately twofold above 5 MM (35.5+/-5.8 vs. 19.6+/-1.6, P < 0.02, n = 3, and 21.0+/-1.3 vs. 10.0+/-1.2 pmol/ 10(6) cells, n = 3, P < 0.001, respectively). In contrast, in glyoxalase-I-transfected cells, 30 mM glucose incubation did not increase MG concentration at all, while increasing the enzymatic product D-lactate by > 10-fold (18.9+/-3.2 vs. 18.4+/- 5.8, n = 3, P = NS, and 107.1+/-9.0 vs. 9.4+/-0 pmol/10(6) cells, n = 3, P < 0.001, respectively). After exposure to 30 mM glucose, intracellular AGE formation in neo cells was increased 13.6-fold (2.58+/-0.15 vs. 0.19+/-0.03 total absorbance units, n = 3, P < 0.001). Concomitant with increased intracellular AGEs, macromolecular endocytosis by these cells was increased 2.2-fold. Overexpression of glyoxalase-I completely prevented both hyperglycemia-induced AGE formation and increased macromolecular endocytosis.
Asunto(s)
Endocitosis , Endotelio Vascular/metabolismo , Endotelio Vascular/fisiología , Productos Finales de Glicación Avanzada/metabolismo , Lactoilglutatión Liasa/metabolismo , Acetona/análogos & derivados , Acetona/metabolismo , Aldehídos/metabolismo , Animales , Bovinos , Endotelio Vascular/enzimología , Regulación Enzimológica de la Expresión Génica , Glucosa/metabolismo , Glutatión/análogos & derivados , Glutatión/farmacología , Glioxal/metabolismo , Hiperglucemia/metabolismo , Cetonas/metabolismo , Ácido Láctico/metabolismo , Lactoilglutatión Liasa/genética , Lisina/análogos & derivados , Lisina/metabolismo , Piruvaldehído/metabolismo , Transfección , Células Tumorales CultivadasRESUMEN
Hyperglycemia rapidly induces an increase in intracellular advanced glycation end products (AGEs) in bovine endothelial cells, causing an alteration in bFGF activity (Giardino, I., D. Edelstein, and M. Brownlee. 1994. J. Clin. Invest. 94:110-117). Because sugar or sugar-adduct autoxidation is critical for AGE formation in vitro, we evaluated the role of reactive oxygen species (ROS) in intracellular AGE formation, using bovine aortic endothelial cells. 30 mM glucose increased intracellular ROS formation by 250% and lipid peroxidation by 330%, while not affecting ROS in the media. In cells depleted of glutathione, intracellular AGE accumulation increased linearly with ROS generation as measured by immunoblotting and the fluorescent probe DCFH (AGE 0.258-3.531 AU* mm/5x10(4) cells, DCF 57-149 mean AU, r = .998, P < .002). Deferoxamine, alpha-tocopherol, and dimethylsulfoxide each inhibited hyperglycemia-induced formation of both ROS and AGE. To differentiate an effect of ROS generation on AGE formation from an effect of more distal oxidative processes, GM7373 endothelial cell lines were generated that stably expressed the peroxidation-suppressing proto-oncogene bcl-2. bcl-2 had no effect on hyperglycemia-induced intracellular ROS formation. In contrast, bcl-2 expression decreased both lipid peroxidation (100% at 3 h and 29% at 168 h) and AGE formation (55% at 168 h). These data show that a ROS-dependent process plays a central role in the generation of intracellular AGEs, and that inhibition of oxidant pathways prevents intracellular AGE formation.
Asunto(s)
Antioxidantes/farmacología , Endotelio Vascular/metabolismo , Glucosa/farmacología , Productos Finales de Glicación Avanzada/biosíntesis , Proteínas Proto-Oncogénicas/biosíntesis , Animales , Bovinos , Células Cultivadas , Antagonismo de Drogas , Hiperglucemia/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2RESUMEN
Intracellular sugars are more reactive glycosylating agents than glucose. In vitro nonezymatic glycosylation of basic fibroblast growth factor (bFGF) by fructose, glucose-6-phosphate (G6P), or glyceraldehyde-3-phosphate (G3P) reduced high affinity heparin-binding activity of recombinant bFGF by 73, 77, and 89%, respectively. Mitogenic activity was reduced 40, 50, and 90%. To investigate the effects of bFGF glycosylation in GM7373 endothelial cells, we first demonstrated that GLUT-1 transporters were not downregulated by increased glucose concentration. In 30 mM glucose, the rate of glucose transport increased 11.6-fold, and the intracellular glucose concentration increased sixfold at 24 h and fivefold at 168 h. The level of total cytosolic protein modified by advanced glycosylation end-products (AGEs) was increased 13.8-fold at 168 h. Under these conditions, mitogenic activity of endothelial cell cytosol was reduced 70%. Anti-bFGF antibody completely neutralized the mitogenic activity at both 5 and 30 nM glucose, demonstrating that all the mitogenic activity was due to bFGF. Immunoblotting and ELISA showed that 30 mM glucose did not decrease detectable bFGF protein, suggesting that the marked decrease in bFGF mitogenic activity resulted from posttranslational modification of bFGF induced by elevated glucose concentration. Cytosolic AGE-bFGF was increased 6.1-fold at 168 h. These data are consistent with the hypothesis that nonenzymatic glycosylation of intracellular protein alters vascular cell function.
Asunto(s)
Diabetes Mellitus/metabolismo , Endotelio Vascular/metabolismo , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Animales , Bovinos , División Celular , Células Cultivadas , Glucosa/farmacología , Transportador de Glucosa de Tipo 1 , Productos Finales de Glicación Avanzada/metabolismo , Glicosilación , Heparina/metabolismo , Humanos , Proteínas de Transporte de Monosacáridos/análisisRESUMEN
In an ophthalmologic study of 90 children with symptomatic giardiasis, ocular alterations were found in 10. Eight of these subjects presented an extensive "salt and pepper" degeneration of the pigmented epithelium involving 360 degrees of the midperiphery of both eyes. In one of the eight children, the pigmented epithelium showed atrophic areas, and in another there was a small hard exudate in the left eye. Of 2 remaining of the 10 children with ocular alterations, 1 presented with slight decoloration of the temporal half of the optic disc, and the other was affected by chorioretinitis. After single-dose antiprotozoic therapy (tinidazole 50 mg/kg), parasitologic tests were negative in all subjects and remained so throughout a 1-year follow-up. However, the characteristic epithelial lesion remained unaltered in all eight children for the entire follow-up period, as well as the optic disc decoloration in the only observed case. The child affected by chorioretinitis recovered after 3 weeks of combined treatment with bethametasone plus deflazacort. In two control groups, 1 of 200 healthy children and 1 of 200 children with gastrointestinal symptoms but without giardiasis, no case of "salt and pepper" degeneration of the pigmented epithelium or other significant ocular alterations was found.