Plasma oxylipin profiling by high resolution mass spectrometry reveal signatures of inflammation and hypermetabolism in amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by progressive loss of motor neurons, systemic hypermetabolism, and inflammation. In this context, oxylipins have been investigated as signaling molecules linked to neurodegeneration, although their specific role in ALS remains unclear. Importantly, most methods focused on oxylipin analysis are based on low-resolution mass spectrometry, which usually confers high sensitivity, but not great accuracy for molecular characterization, as provided by high-resolution MS (HRMS). Here, we established an ultra-high performance liquid chromatography HRMS (LC-HRMS) method for simultaneous analysis of 126 oxylipins in plasma. Intra- and inter-day method validation showed high sensitivity (0.3-25 pg), accuracy and precision for more than 90% of quality controls. This method was applied in plasma of ALS rats overexpressing the mutant human Cu/Zn-superoxide dismutase gene (SOD1-G93A) at asymptomatic (ALS 70 days old) and symptomatic stages (ALS 120 days old), and their respective age-matched wild type controls. From the 56 oxylipins identified in plasma, 17 species were significantly altered. Remarkably, most of oxylipins linked to inflammation and oxidative stress derived from arachidonic acid (AA), like prostaglandins and mono-hydroxides, were increased in ALS 120 d rats. In addition, ketones derived from AA and linoleic acid (LA) were increased in both WT 120 d and ALS 120 d groups, supporting that age also modulates oxylipin metabolism in plasma. Interestingly, the LA-derived diols involved in fatty acid uptake and ß-oxidation, 9(10)-DiHOME and 12(13)-DiHOME, were decreased in ALS 120 d rats and showed significant synergic effects between age and disease factors. In summary, we validated a high-throughput LC-HRMS method for oxylipin analysis and provided a comprehensive overview of plasma oxylipins involved in ALS disease progression. Noteworthy, the oxylipins altered in plasma have potential to be investigated as biomarkers for inflammation and hypermetabolism in ALS.

Electrophilic oxysterols: generation, measurement and protein modification.

Cholesterol is an essential component of mammalian plasma membranes. Alterations in sterol metabolism or oxidation have been linked to various pathological conditions, including cardiovascular diseases, cancer, and neurodegenerative disorders. Unsaturated sterols are vulnerable to oxidation induced by singlet oxygen and other reactive oxygen species. This process yields reactive sterol oxidation products, including hydroperoxides, epoxides as well as aldehydes. These oxysterols, in particular those with high electrophilicity, can modify nucleophilic sites in biomolecules and affect many cellular functions. Here, we review the generation and measurement of reactive sterol oxidation products with emphasis on cholesterol hydroperoxides and aldehyde derivatives (electrophilic oxysterols) and their effects on protein modifications.

LACK OF ASSOCIATION BETWEEN HERPESVIRUS DETECTION IN SALIVA AND GINGIVITIS IN HIV­INFECTED CHILDREN.

The aims of this study were to compare the detection of human herpesviruses (HHVs) in the saliva of HIV-infected and healthy control children, and to evaluate associations between viral infection and gingivitis and immunodeficiency. Saliva samples were collected from 48 HIV-infected and 48 healthy control children. Clinical and laboratory data were collected during dental visits and from medical records. A trained dentist determined gingival indices and extension of gingivitis. Saliva samples were tested for herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), and cytomegalovirus (CMV) by nested polymerase chain reaction assays. Thirty-five HIV-infected and 16 control children had gingivitis. Seventeen (35.4%) HIV-infected children and 13 (27%) control children were positive for HHVs. CMV was the most commonly detected HHV in both groups (HIV-infected, 25%; control, 12.5%), followed by HSV-1 (6.2% in both groups) and HSV-2 (HIV-infected, 4.2%; control, 8.3%). The presence of HHVs in saliva was not associated with the presence of gingivitis in HIV-1-infected children (p = 0.104) or healthy control children (p = 0.251), or with immunosuppression in HIV-infected individuals (p = 0.447). Gingivitis was correlated with HIV infection (p = 0.0001). These results suggest that asymptomatic salivary detection of HHVs is common in HIV-infected and healthy children, and that it is not associated with gingivitis.

Lack of associaiion between herpesvirus detectin in saliva and gingivitis in hhiv‑infected children / Ausência de associação entre a detecção de herpesvírus na saliva e gengivite em crianças infectadas pelo HIV

The aims of this study were to compare the detection of human herpesviruses (HHVs) in the saliva of HIV-infected and healthy control children, and to evaluate associations between viral infection and gingivitis and immunodeficiency. Saliva samples were collected from 48 HIV-infected and 48 healthy control children. Clinical and laboratory data were collected during dental visits and from medical records. A trained dentist determined gingival indices and extension of gingivitis. Saliva samples were tested for herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), varicella zoster virus (VZV), Epstein-Barr virus (EBV), and cytomegalovirus (CMV) by nested polymerase chain reaction assays. Thirty-five HIV-infected and 16 control children had gingivitis. Seventeen (35.4%) HIV-infected children and 13 (27%) control children were positive for HHVs. CMV was the most commonly detected HHV in both groups (HIV-infected, 25%; control, 12.5%), followed by HSV-1 (6.2% in both groups) and HSV-2 (HIV-infected, 4.2%; control, 8.3%). The presence of HHVs in saliva was not associated with the presence of gingivitis in HIV-1-infected children (p = 0.104) or healthy control children (p = 0.251), or with immunosuppression in HIV-infected individuals (p = 0.447). Gingivitis was correlated with HIV infection (p = 0.0001). These results suggest that asymptomatic salivary detection of HHVs is common in HIV-infected and healthy children, and that it is not associated with gingivitis.

Os objetivos deste estudo foram detectar a presença de herpesvírus humanos (HHVs) na saliva de crianças infectadas pelo HIV, em comparação com controles saudáveis e avaliar a associação entre infecção viral, gengivite e imunodeficiência. Para este fim, foram colhidas amostras de saliva de 48 crianças HIV-positivas e 48 controles saudáveis. O índice gengival e extensão de gengivite foram determinados por um dentista treinado. Informações clínicas e laboratoriais foram obtidas durante a consulta odontológica e dos registros médicos. As amostras de saliva foram testadas para detecção de vírus herpes simplex tipos 1 e 2 (HSV-1 e HSV-2), vírus da varicela-zoster (VVZ), vírus Epistein-Barr (EBV) e citomegalovírus (CMV) através de nested-PCR. Trinta e cinco crianças HIV-positivas e 16 crianças do grupo controle apresentavam gengivite. Dezessete (35,4%) crianças HIV-positivas e 13 (27%) crianças controle testaram positivo para a presença de HHVs. CMV foi o vírus mais comum detectado em ambos os grupos (25% HIV-positivas e 12,5% de controle), seguido por HSV-1 (6,2% de ambos os grupos) e HSV-2 (4,2% HIV-positivas e 8,3% de controle). Não houve associação entre a detecção de HHVs na saliva e a presença de gengivite em ciranças HIV-positivas (p = 0.104) ou crianças saudáveis (p = 0,251), ou com imunossupressão em indivíduos HIV-positivos (p = 0,447). Foi observada uma correlação entre a infecção por HIV e a presença de gengivite (p = 0,0001). Os resultados sugerem que a detecção salivar assintomática de HHVs é comum entre crianças HIV-positivas e crianças saudáveis, e não está associada à gengivite.

Multiple-gene characterization of rotavirus strains: evidence of genetic linkage among the VP7-, VP4-, VP6-, and NSP4-encoding genes.

A total of 162 rotavirus strains detected between 1996 and 2006 among individuals with diarrhea in Rio de Janeiro, Brazil, were analyzed by multiple-gene genotyping. Characterization of strains was done by RT-PCR assay for amplification and typing of the VP7-, VP4-, VP6-, and NSP4-encoding genes. Overall, 139 (85.8%) strains belonged to the common group A rotavirus combinations: 67 (41.4%) belonged to genotype G1-P[8]-I1-E1; 18 (11.1%) were G2-P[4]-I2-E2; 11 (6.8%) were G3-P[8]-I1-E1; 12 (7.4%) were G4-P[8]-I1-E1; and 31 (19.1%) were G9-P[8]-I1-E1. Two samples presented mixed genotypes (G1 + G3-P[8]-I1-E1 and G1 + G9-P[9]-I1-E1) and rare combinations, such as G2-P[6]-I2-E2 and G9-P[6]-I2-E2, were detected in six (3.7%) strains. The results suggest a linkage among all four genes. Genotypes G1/G3/G4/G5/G9-P[8] were correlated strongly to I1-E1 genotypes and G2-P[4]/P[6] were correlated to I2-E2 genotypes. Unusual combinations of genes, such as G3-P[9]-I2-E2, G9-P[9]-I1-E2, and G3-P[9]-I3-E3, were observed in 15 (9.3%) strains. The characterization of multiple genes allows a more complete analysis of the rotavirus isolates and provides evidence of natural reassortment of strains.

Validade dos esfigmomanômetros utilizados por profissionais de saúde do hospital universitário da Universidade de Brasília / Validity of the sphygmomanometers used by health care professionals at the university hospital of the University of Brasilia

Objetivo: Determinar a precisão dos esfigmomanômetros aneróides, utilizados no Hospital Universitário da Universidade de Brasília. Material e métodos: Foram avaliados 57 aparelhos em relação a defeitos físicos e à precisão. Em novembro de 1997, dois técnicos do Instituto Nacional de Metrologia, Normalização e Qualidade Industrial (Inmetro) aferiram os esfigmomanômetros aneróides portáteis de médicos, enfermeiros e alunos do Hospital Universitário da Universidade de Brasília, utilizando-se o equipamento Onneken-Calibrator, modelo OM 631.000 S (Alemanha). Resultados: O defeito mais encontrado, na avaliação de 12 equipamentos, foi a perda do batente limitador (7/12). Outros defeitos foram vazamento de ar (2/12) e visor estragado (3/12). Esses aparelhos foram reprovados na aferição. Pelos critérios do Inmetro, 35 aparelhos foram aprovados e receberam o selo de validade, e 10 aparelhos foram reprovados. Se considerarmos os critérios do National Bureau of Standards e os da Association for the Advancement of Medical Instrumentation, 15 aparelhos teriam sido reprovados, dos quais nove superestimaram a medida da pressão sanguínea arterial e seis a subestimaram. Dos 57 aparelhos, 55 (96,5 por cento) estavam em uso. Conclusão: Constatou-se que 22 (38,5 por cento) de todos os manômetros avaliados, segundo os critérios do Inmetro, apresentaram-se inadequados à pratica clínica, 12 (21 por cento) por defeitos físicos e 10 (17,5 por cento) por imprecisão. Esses números estão de acordo com os descritos na literatura. Se considerarmos os critérios mais rígidos do National Bureau Standards da Association for the Advancement of Medical Instrumentation, essa porcentagem subirá para 47 por cento.