A propósito de un caso: micosis pulmonar en paciente pediátrico con enfermedad granulomatosa crónica / Case report: pulmonary mycosis in children with chronic granulomatous diseaseSE

A 8-year-old girl, born and resident of Guárico state, was referred to our Department because of a history of recurrent pneumonia and a history of deceased sister by unspecified lung disease. At the age of 4 years she had suffered several episodes of pneumonia requiring hospitalization. These episodes were no-responsive to antibiotic therapy and treatment for tuberculosis. Subsequently, At the age of 8 years she was hospitalized again for an episode of left lower lobe pneumonia that did not improve. Studies were performed to rule out pulmonary pathology disease: Cystic Fibrosis was discarded and also Pulmonary Tuberculosis. Serology for HIV and Fungi were negative. Because serological studies were inconclusive, a videobronchoscopy plus Bronchoalveolar lavage and lung tissue biopsy were performed, which reported bronchitis and chronic granulomatous and caseous necrosis. Special stains were observed that suggest fungus infection. Primary immunodeficiency was suspected in the patient, because the presence of recurrent pneumonia of different etiologies. The presence of the granuloma observed by the videobronchoscopy. A positive culture for Histoplasma and Aspergillus fungi, and the result of the oxidative capacity test, where the deficiency was observed in the microbicidal activity of macrophages. They were strong evidence that corroborated the immunodeficiency called Chronic Granulomatous Disease.

Escolar femenino de 8 años de edad, natural y procedente del Estado Guárico, con antecedente de hospitalizaciones por neumonías recurrente desde los 4, recibió antibioticoterapia endovenosa y cumplió tratamiento antifímico en dos oportunidades, persistiendo con sintomatología respiratoria. A los 8 años precisó nueva hospitalización por diagnóstico de neumonía del lóbulo inferior izquierdo. Por no presentar mejoría y antecedente de hermana fallecida por patología pulmonar no precisada fue referida a nuestro centro. Se realizaron estudios por patología pulmonar crónica: se descartó Fibrosis Quística y Tuberculosis Pulmonar. Serología para HIV y Hongos Negativa. Por no ser concluyentes los estudios serológicos se realizó Videobroncoscopia más lavado y biopsia, la cual reportó bronquitis crónica granulomatosa y necrosis caseosa. Coloraciones especiales: hongos intracitoplasmáticos sugestivos de Histoplasma Sp. y en el cultivo presentó crecimiento de Aspergillus fumigatus. Ante la presencia de paciente con neumonía recurrente por diferentes etiologías se sospechó la presencia de Inmunodeficiencia Primaria, planteando en base al reporte de la videobroncoscopia de granuloma y la confirmación de Infección por Histoplasma y Aspergillus una Enfermedad Granulomatosa Crónica que fue documentada al medir la deficiencia en la actividad microbicida dependiente de oxigeno evaluada a través del Test de Capacidad Oxidativa.

Early involvement of ROS overproduction in apoptosis induced by 7-ketocholesterol.

Cholesterol oxidation products are increasingly considered as much more bioactive than the parent compound in the multifactor and multistep process that characterizes atherosclerosis. In particular, 7-ketocholesterol has been reported to induce oxidative stress as well as a marked pro-apoptotic effect in vascular cells including macrophages. With the aim to investigate a possible pathogenic correlation between the two events, cultivated murine macrophages were challenged with a concentration of 7-ketocholesterol actually detectable in human vasculature. Conclusive proof was obtained of a primary role of NADPH-oxidase in the overproduction of reactive oxygen species within cells treated with the oxysterol. In addition, such oxidative burst occurred very early after cell intoxication and it was definitely demonstrated as able to lead cells to apoptotic death. In fact, two metabolic inhibitors of NADPH-oxidase and the antioxidant epicatechin very well counteracted 7-ketocholesterol-induced apoptosis by preventing the oxysterol pro-oxidant action.

Oxidative status in chronic hepatitis C: the influence of antiviral therapy and prognostic value of serum hydroperoxide assay.

UNLABELLED: The effect of alpha-interferon (alpha-IFN) and ribavirin (RBV) treatment on oxidative status in chronic hepatitis C (CHC) is unknown. AIM: To study the time course of oxidative status in patients with CHC during alpha-IFN and RBV administration, and to evaluate the role of oxidative status in order to predict the therapeutic response. PATIENTS AND METHODS: Fifty one patients with CHC were studied. All received a combination of alpha-IFN and RBV for 6 or 12 months in relation to the type of response. The hydroperoxides concentration in serum test samples by D-ROM test was measured in all of the patients before therapy. In 27 patients, hydroperoxides were also measured during the treatment and during the 12 subsequent months. RESULTS: Cross-sectional analysis demonstrates that patients with a successive long-term response had a lower basal serum hydroperoxide concentration than non-responders (280 +/- 40.8 vs 337 +/- 83 CARR Units, p < 0.05). This resulted to be an independent factor predictive of long-term response in the multi-varied analysis. Longitudinal observation on 27 patients showed that the mean hydroperoxide concentration decreased significantly during treatment (T0 329 +/- 79.2 vs T12 272 +/- 34.5 CARR Units) and that the decrease in the mean values was mainly due to variations in the relapsers group. CONCLUSIONS: Normal basal hydroperoxide concentration helps to predict long-term response to combination therapy. The D-ROM test may be used for screening patients before treatment.

Oxysterol mixtures prevent proapoptotic effects of 7-ketocholesterol in macrophages: implications for proatherogenic gene modulation.

Oxysterols are common components of oxidized low-density lipoprotein and accumulate in the core of fibrotic plaques as a mixture of cholesterol and cholesteryl ester oxidation products. The proapoptotic effects of a biologically representative mixture of oxysterols was compared with equimolar amounts of 7-ketocholesterol and unoxidized cholesterol. The oxysterol mixture in a concentration range actually detectable in hypercholesterolemic patients did not stimulate programmed cell death in cultivated murine macrophages. Unoxidized cholesterol also produced no effect. By contrast, when given alone, 7-ketocholesterol strongly stimulated the mitochondrial pathway of apoptosis with cytochrome c release, caspase-9 activation, and eventually caspase-3 activation. Subsequent experiments showed that when 7-ketocholesterol was administered to cells together with another oxysterol, namely 7betaOH-cholesterol, the strong proapoptotic effect of 7-ketocholesterol was markedly attenuated. As regards the mechanism underlying this quenching, we found that the combined oxysterol treatment counteracted the ability of 7-ketocholesterol, when administered alone, to strongly up-regulate the steady-state levels of reactive oxygen species (ROS) without interfering with sterol uptake. Furthermore, this increase in intracellular ROS appeared to be responsible for the up-regulation of proapoptotic factor, p21, after treatment with 7-ketocholesterol but not in cells challenged with the oxysterol mixture. Competition among oxysterols, apparently at the level of NADPH oxidase, diminishes the ROS induction and direct toxicity that is evoked by specific oxysterols. As a consequence, a more subtle gene modulation by oxysterols becomes facilitated in vascular cells.

Use of N,N-dimethyl-p-phenylenediamine to evaluate the oxidative status of human plasma.

Oxidative stress has been clearly implicated in human disease by a growing body of scientific evidences. There is no ideal method for the measurement of this parameter. A possible strategy would be to measure simultaneously several biomarkers representing damage to different cellular components or, alternatively, a method able to evaluate the hydroperoxides, intermediate products of oxidation originating from different classes of molecules, such as lipids, peptides, amino acids, etc. can be used. We are introducing a simple, rapid and inexpensive assay to measure the oxidative status of human plasma. It is based on the properties of N,N-dimethyl-p-phenylenediamine (DMPD), a compound able to produce a fairly long-lived radical cation. The absorbance at 505 nm of a DMPD solution in the presence of plasma, which is proportional to the amount of hydroperoxyl compounds, is related to the oxidative status of the sample and could be expressed as hydrogen peroxide equivalents (HPE). This assay was not influenced by freezing-thawing and storage time of the plasma samples. The assay can be automated, performed in a kinetic mode, and used for routine analyses. The DMPD assay alone or in combination with analytical methods for assessing antioxidant capacity is suggested as a reliable tool to obtain information in pathologies related to oxidative stress.