Untargeted LC-HRMS metabolomics reveals candidate biomarkers for mucopolysaccharidoses.

BACKGROUND: Mucopolysaccharidoses (MPSs) are inherited genetic diseases caused by an absence or deficiency of lysosomal enzymes responsible for catabolizing glycosaminoglycans (GAGs). Undiagnosed patients, or those without adequate treatment in early life, can be severely and irreversibly affected by the disease. In this study, we applied liquid chromatography-high resolution mass spectrometry (LC-HRMS)-based untargeted metabolomics to identify potential biomarkers for MPS disorders to better understand how MPS may affect the metabolome of patients. METHODS: Urine samples from 37 MPS patients (types I, II, III, IV, and VI; untreated and treated with enzyme replacement therapy (ERT)) and 38 controls were analyzed by LC-HRMS. Data were processed by an untargeted metabolomics workflow and submitted to multivariate statistical analyses to reveal significant differences between the MPS and control groups. RESULTS: A total of 12 increased metabolites common to all MPS types were identified. Dipeptides, amino acids and derivatives were increased in the MPS group compared to controls. N-acetylgalactosamines 4- or 6-sulfate, important constituents of GAGs, were also elevated in MPS patients, most prominently in those with MPS VI. Notably, treated patients exhibited lower levels of the aforementioned acylaminosugars than untreated patients in all MPS types. CONCLUSIONS: Untargeted metabolomics has enabled the detection of metabolites that could improve our understanding of MPS physiopathology. These potential biomarkers can be utilized in screening methods to support diagnosis and ERT monitoring.

Early hematopoietic stem cell transplantation in a patient with severe mucopolysaccharidosis II: A 7 years follow-up.

Mucopolysaccharidosis type II (MPS II - Hunter syndrome) is an X-linked lysosomal storage disorder caused by a deficiency in the enzyme iduronate-2 sulfatase (I2S), leading to the accumulation of the glycosaminoglycans, affecting multiple organs and systems. Enzyme replacement therapy does not cross the blood brain barrier, limiting results in neurological forms of the disease. Another option of treatment for severe MPS, hematopoietic stem cell transplantation (HSCT) has become the treatment of choice for the severe form of MPS type I, since it can preserve neurocognition when performed early in the course of the disease. To date, only few studies have examined the long-term outcomes of HSCT in patients with MPS II. We describe the seven-year follow-up of a prenatally diagnosed MPS II boy with positive family history of severe MPS form, submitted to HSCT with umbilical cord blood cells at 70 days of age. Engraftment after 30 days revealed mixed chimerism with 79% donor cells; after 7 years engraftment remains at 80%. I2S activity 30 days post-transplant was low in plasma and normal in leukocytes and the same pattern is observed to date. At age 7 years growth charts are normal and he is very healthy, although mild signs of dysostosis multiplex are present, as well as hearing loss. The neuropsychological evaluation (Wechsler Intelligence Scale for Children - Fourth Edition - WISC-IV), disclosed an IQ of 47. Despite this low measured IQ, the patient continues to show improvements in cognitive, language and motor skills, being quite functional. We believe that HSCT is a therapeutic option for MPS II patients with the severe phenotype, as it could preserve neurocognition or even halt neurodegeneration, provided strict selection criteria are followed.

O Cinismo como Metáfora da Cultura / Cynicism as a Metaphor of Culture / El Cinismo como Metáfora de la Cultura / Le Cynisme comme Metaphore de la Culture

Este trabalho teve como objetivo analisar o cinismo enquanto metáfora da cultura na atualidade. Buscamos estudá-lo em seus aspectos histórico-sociais e em suas relações com a economia libidinal dos sujeitos. Trata-se de pesquisa teórica, fundamentada na Psicanálise, desde Freud e em Fábio Herrmann, autor que nos permitiu realizar a recuperação do método psicanalítico na análise da cultura contemporânea. A partir das leituras realizadas, verificamos que a racionalidade cínica é modo de funcionamento das estruturas de sociabilidade que aparecem em processos de crise de legitimação e erosão das normas da vida social. O desempenho erótico das subjetividades encontra-se alinhado com as condições econômicas e sociais, o que implica em uma nova economia libidinal dos sujeitos, no âmbito do capitalismo cínico.

This work aimed at analyzing cynicism as a metaphor of contemporary culture. We seek to study it in its historical-social aspects and in its relations with the subjects' libidinal economy. It is a theoretical research, based on Psychoanalysis, from Freud and Fábio Herrmann, the author who allowed us to realize the recovery of the psychoanalytic method in the analysis of contemporary culture. From the readings, we verify that cynical rationality is the mode of functioning of the structures of sociability that appear in processes of a crisis of legitimation and erosion of norms of social life. The erotic performance of subjectivities is aligned with economic and social conditions, which implies a new libidinal economy of the subjects, within the framework of cynical capitalism.

El objetivo de este trabajo fue analizar el cinismo como metáfora de la cultura en la actualidad. Buscamos estudiarlo en sus aspectos histórico-sociales y en sus relaciones con la economía libidinal de los sujetos. Se trata de una investigación teórica, basada en la Psicoanálisis, desde Freud y en Fábio Herrmann, autor que nos permitió realizar la recuperación del método psicoanalítico en el análisis de la cultura contemporánea. De las lecturas realizadas, verificamos que la racionalidad cínica es el modo de funcionamiento de las estructuras de sociabilidad que surgen en procesos de crisis de legitimación y erosión de las reglas de la vida social. El rendimiento erótico de las subjetividades se encuentra alineado a las condiciones económicas y sociales, lo que implica en una nueva economía libidinal de los sujetos, en el ámbito del capitalismo cínico.

Ce travail a eu l'objectif d'analyser le cynisme comme métaphore de la culture aujourd'hui. Nous recherchons étudier le cynisme dans ses aspects historiques et sociaux et dans ses relations avec l'économie libidinale des sujets. C'est une recherche théorique, basée dans la Psychanalyse, depuis Freud et à Fábio Herrmann, auteur qui nous a permis de réaliser la récupération de la méthode psychanalytique dans l'analyse de la culture contemporaine. D'après les lectures realisées, nous avons verifié que la rationalité cynique est une façon de fonctionnement des structures de sociabilité qui apparaissent dans les processus de crise de légitimité et de l'érosion des normes de la vie sociale. La performance érotique des subjectivités est alignée avec les conditions économiques et sociales, cela implique une nouvelle économie libidinale des sujets, dans le cadre du capitalisme cynique.

Systematic analysis of glycerol: colourimetric screening and gas chromatography-mass spectrometric confirmation.

Glycerol is a naturally occurring polyol in the human body, essential for several metabolic processes. It is widely used in the food, pharmaceutical, and medical industries and in clinical practice as a plasma volume expander (PVE). Athletes, however, may use glycerol to mask the presence of forbidden substances or to enhance performance, inclusively through hyperhydration achieved by glycerol ingestion with added fluid. These practices are considered doping, and are prohibited by the World Anti-Doping Agency (WADA). Therefore, glycerol was introduced in the prohibited list. Doping through glycerol ingestion can readily be identified by detection of elevated glycerol concentrations in urine. In this paper, a protocol for the fast detection of glycerol in urine is proposed. It consists of a previous visual colourimetric screening, followed by a quantitative/qualitative confirmation analysis by mass spectrometry. The screening procedure involves a reaction in which polyhydric alcohols are oxidized by periodate to formic acid and formaldehyde, which is detected by the addition of a fuchsin solution. For the subsequent qualitative/quantitative confirmation analysis, a gas chromatography-mass spectrometry based approach with a non-deuterated internal standard and a drying step of only 10 min is proposed. The linear correlation was demonstrated within WADA´s threshold range. The calculated RSD were 2.1% for within-day precision and 2.8% for between-day precision. The uncertainty estimation was calculated, and a value of 2.7% was obtained. The procedure may also be used for the analysis of other polyols in urine, as for example the PVE mannitol.

Percepção dos profissionais de saúde e comunitários em relação à terapia comunitária na estratégia saúde da família / Perception of health professionals and community members regarding community therapy in the family health strategy

A presente pesquisa tem como objetivo analisar as opiniões dos profissionais de saúde e de usuários em relação à Terapia Comunitária Integrativa na Estratégia Saúde da Família. Trata-se de um estudo descritivo de natureza qualitativa, em uma Unidade de Saúde da Família no Município de Jaboatão dos Guararapes/PE. Os sujeitos da pesquisa foram Profissionais de Saúde que trabalham ou trabalharam na USF ? Unidade de Saúde da Família e Comunitários da área adstrita que participaram de pelo menos um encontro ou roda de TCI. Utilizou-se como instrumento de coleta a entrevista semiestruturada, específica para cada grupo, com análise de narrativa de conteúdo. A TCI promoveu mudanças no ato de interagir, criou laços de amizade, de afeto, de respeito entre profissionais, usuários e comunidade. Para os profissionais trouxe modificações na qualidade da assistência, auxiliando na desconstrução da lógica da cura para a criação da lógica do cuidado. Os achados obtidos nesse estudo permitem sugerir que a Terapia Comunitária Integrativa melhora a qualidade da assistência na Estratégia da Saúde da Família por promover nas pessoas, independente do papel que ela esteja assumindo, o resgate da essência humana, promove aproximação, o cuidar do outro, favorece o ambiente para a escuta, compreensão e visão integral. O sentar em roda evidencia a potencialidade e a força que a comunidade tem quando se encontra unida.

The intent of this research is to analyze health professionals? and users? opinions regarding the Integrative Community Therapy (ICT) in the Family Health Strategy. This is a descriptive qualitative study at a Family Health Unit in the city of Jaboatão dos Guararapes/PE. The subjects of the research were health professionals that work or have worked at the FHU - Family Health Unit, and the Community members in its service area who have taken part in at least one ICT meeting or circle. A semi-structured interview, specific to each group, was used as a collection instrument, along with a narrative analysis of content. ICT promoted changes in the interaction process, created friendly relationships, affection, and respect between professionals, users, and community. For professionals it brought modifications in the quality of assistance, changing the vision from a focus on cure to a focus on care. The results obtained in this study give evidence to suggest that Integrative Community Therapy improves the quality of the assistance in the Family Health Strategy by promoting closer personal ties, taking care of others, and fosters an environment for more comprehensive listening, understanding, and vision. The act of sitting in a circle shows the potential and the force that the community has when it?s united.

Determination of six pterins in urine by LC-MS/MS.

BACKGROUND: The present work describes an analytical method for urinary pterins by LC-MS/MS, with emphasis on the separation of 6- and 7-positional isomers of bio- and neopterins. RESULTS: Urine sample preparation consisted of oxidation by MnO(2), filtration and direct dilution in the mobile phase. The method was validated in urine spiked at five concentration levels with true triplicates of each level. Separation of the pterins, including the positional isomers, was achieved by employing a LUNA amino column. Six pterins were quantified (pterin, isoxanthopterin, 6-biopterin, 7-biopterin, 6-neopterin, 7-neopterin) and a linear behavior was observed; LOD varied from 7 to 360 pg/ml and correlation coefficients above 0.98 were obtained for all pterins. In addition, pterin levels were evaluated in 41 urine samples of healthy subjects, in ten urine samples of patients with classical phenylketonuria (PKU) and in one with atypical PKU. CONCLUSION: The proposed method allowed to identify, separate and quantify six pterins in urine, using a simple and rapid sample preparation. The atypical PKU was unequivocally differentiated from the classical form, demonstrating that this method could be very useful for characterization and follow-up of diseases.

Síndromes hipertensivas graves – estudo descritivo com adolescentes atendidas em maternidade escola / Severe hypertensive syndrome – descriptive study with adolescents attended at a maternity school

Estudo exploratório e descritivo, com o objetivo de analisar os dados clínicos e obstétricos relacionados às síndromes hipertensivas graves ocorridas em adolescentes assistidas em uma maternidade-escola do Recife-PE. A população foi constituída por 186 gestantes adolescentes com diagnóstico de pré-eclâmpsia grave e/ou eclâmpsia ocorrida entre 2003 a 2008. A idade variou entre 15 e 19 anos, eram negras, solteiras e com baixa escolaridade. A maioria era primigesta, mas a recorrência de gestação configurou-se em 16% dos casos. Realizaram seis ou mais consultas de pré-natal, a gestação evoluiu a termo e o tipo de parto mais frequente foi a cesárea. As comorbidades identificadas foram alterações do volume do líquido amniótico, quadros hemorrágicos e infecções. Foram também identificados casos de retardo do crescimento intrauterino, prematuridade, icterícia, hipóxia e baixo peso ao nascer. Concluiu-se que a gravidez na adolescência associada às síndromes hipertensivas graves está relacionada a severas complicações maternas, fetais e neonatais.

Rabies diagnosis and serology in bats from the State of São Paulo, Brazil.

INTRODUCTION: Bats are one of the most important reservoirs and vectors of the rabies virus in the world. METHODS: From 1988 to 2003, the Zoonosis Control Center in São Paulo City performed rabies diagnosis on 5,670 bats by direct immunofluorescent test and mouse inoculation test. Blood samples were collected from 1,618 bats and the sera were analyzed using the rapid fluorescent focus inhibition test to confirm rabies antibodies. RESULTS: Forty-four (0.8%) bats were positive for rabies. The prevalence of rabies antibodies was 5.9% using 0.5IU/ml as a cutoff. Insectivorous bats (69.8%) and bats of the species Molossus molossus (51.8%) constituted the majority of the sample; however, the highest prevalence of antibodies were observed in Glossophaga soricina (14/133), Histiotus velatus (16/60), Desmodus rotundus (8/66), Artibeus lituratus (5/54), Nyctinomops macrotis (3/23), Tadarida brasiliensis (3/48), Carollia perspicillata (3/9), Eumops auripendulus (2/30), Nyctinomops laticaudatus (2/16), Sturnira lilium (2/17) and Eumops perotis (1/13). The prevalence of rabies antibodies was analyzed by species, food preference and sex. CONCLUSIONS: The expressive levels of antibodies associated with the low virus positivity verified in these bats indicate that rabies virus circulates actively among them.

Rabies diagnosis and serology in bats from the State of São Paulo, Brazil / Diagnóstico e sorologia de raiva em morcegos do Estado de São Paulo, Brasil

INTRODUCTION: Bats are one of the most important reservoirs and vectors of the rabies virus in the world. METHODS: From 1988 to 2003, the Zoonosis Control Center in São Paulo City performed rabies diagnosis on 5,670 bats by direct immunofluorescent test and mouse inoculation test. Blood samples were collected from 1,618 bats and the sera were analyzed using the rapid fluorescent focus inhibition test to confirm rabies antibodies. RESULTS: Forty-four (0.8 percent) bats were positive for rabies. The prevalence of rabies antibodies was 5.9 percent using 0.5IU/ml as a cutoff. Insectivorous bats (69.8 percent) and bats of the species Molossus molossus (51.8 percent) constituted the majority of the sample; however, the highest prevalence of antibodies were observed in Glossophaga soricina (14/133), Histiotus velatus (16/60), Desmodus rotundus (8/66), Artibeus lituratus (5/54), Nyctinomops macrotis (3/23), Tadarida brasiliensis (3/48), Carollia perspicillata (3/9), Eumops auripendulus (2/30), Nyctinomops laticaudatus (2/16), Sturnira lilium (2/17) and Eumops perotis (1/13). The prevalence of rabies antibodies was analyzed by species, food preference and sex. CONCLUSIONS: The expressive levels of antibodies associated with the low virus positivity verified in these bats indicate that rabies virus circulates actively among them.

INTRODUÇÃO: Morcegos são um dos mais importantes reservatórios e vetores do vírus da raiva no mundo. MÉTODOS: No período entre 1998 e 2003, o Centro de Controle de Zoonoses da Cidade de São Paulo realizou o diagnóstico de raiva em 5.670 morcegos utilizando as técnicas de imunofluorescência direta e inoculação intracerebral em camundongos. Sangue foi coletado de 1.618 espécimes para pesquisa de anticorpos pela técnica de inibição de foco de fluorescência rápida. RESULTADOS: Quarenta e quatro (0,8 por cento) morcegos foram positivos para raiva. A prevalência de anticorpos foi de 5,9 por cento usando 0,5UI/ml como ponto de corte. Os morcegos de hábito alimentar insetívoro (69,8 por cento) e os morcegos da espécie Molossus molossus (51,8 por cento) representaram a maioria da amostra. Entretanto, as mais altas prevalências de anticorpos foram observadas nos morcegos Glossophaga soricina (14/133), Histiotus velatus (16/60), Desmodus rotundus (8/66), Artibeus lituratus (5/54), Nyctinomops macrotis (3/23), Tadarida brasiliensis (3/48), Carollia perspicillata (3/9), Eumops auripendulus (2/30), Nyctinomops laticaudatus (2/16), Sturnira lilium (2/17) e Eumops perotis (1/13). A prevalência de anticorpos foi analisada por espécie, hábito alimentar e sexo. CONCLUSÕES: O expressivo nível de anticorpos associado à baixa positividade para o vírus da raiva entre os morcegos estudados indica que o vírus circula ativamente entre morcegos.

Plasma volume expanders: use in medicine and detecting misuse in sports.

Plasma volume expanders comprise a heterogeneous group of substances used in medicine that are intravenously administered in cases of great blood loss owing to surgery or medical emergency. These substances, however, can also be used to artificially enhance performance of healthy athletes in sport activities, and to mask the presence of others substances. These practices are considered doping, and are therefore prohibited by the International Olympic Committee and the World Antidoping Agency. Consequently, drug testing procedures are essential. The present work provides an overview of plasma volume expanders, assembling pertinent data such as chemical characteristics, physiological aspects, adverse effects, doping and analytical detection methods, which are currently dispersed in the literature.

Oral exfoliative cytology for the diagnosis of paracoccidoidomycosis in a patient with human immunodeficiency virus: a case report.

BACKGROUND: Paracoccidioidomycosis is not the most common fungal disease in patients infected with human immunodeficiency virus (HIV), except for endemic regions in Latin America countries. CASE: A 33-year-old man with HIV presented with mulberry-like lesions on the palate. The diagnosis was made by exfoliative cytology and Papanicolaou staining. Microscopic analysis revealed fungal structures with birefringent walls and exosporulation conferring an airplane radial motor appearance, or even bowel-like or goblet-like forms compatible with Paracoccidioides brasiliensis. CONCLUSION: This process spares the immunosuppressed patient from undergoing invasive biopsy procedures.

Detection of Leishmania (Leishmania) amazonensis and Leishmania (Leishmania) infantum chagasi in Brazilian bats.

Although bats are one of the most abundant mammals in the new world and are present in virtually all ecosystems, including urban and peri-urban environments, few studies have investigated the role of these animals in the epidemiological chain of leishmaniosis. Here, we report a study of 683 bats captured in São Paulo county (southeastern from Brazil), which were screened by serology, parasitologic methods and polymerase chain reaction (PCR) for trypanosomatids. The indirect immunofluorescent antibody test demonstrated that 0.9% of bats react positively for leishmaniosis and PCR detected the presence of DNA of Leishmania (Leishmania) amazonensis in 18 bats and Leishmania (Leishmania) infantum chagasi in 3 specimens. These results indicate that further studies are necessary to evaluate the role of bats in maintenance of the Leishmania life cycle, especially in areas where these diseases are endemic.

Screening for hydroxyethyl starch (HES) doping in sport.

The artificial colloid hydroxyethyl starch (HES) is among the most frequently used plasma volume expanders in the medical field. However, in 1998, its misuse by the athletic community was officially reported and since 2000, HES is prohibited by the International Olympic Committee (IOC). Therefore, several methods enabling the detection of HES in urine were developed, most based on gas chromatography-mass spectrometry (GC-MS). In the present work, a simple and low-cost screening method, intended to reduce the number of samples to be analysed by GC-MS, was developed. The method is based on the acid hydrolysis of HES and detection of the resulting glucose and hydroxyethyl glucose derivatives by Benedict's reaction (reduction of copper sulfate to brick-red cuprous oxide by glucose and/or derivatives). Samples considered suspect were submitted to GC-MS analysis for identification of HES. The method was successfully applied for screening of HES in 2627 urine samples from 1346 Brazilian soccer players and 1281 athletes from the Pan-American Games (Rio de Janeiro, 2007); 71 (2.7%) samples, considered suspect, were submitted to GC-MS, but no positive results were found. Moreover, a thin layer chromatography (TLC) method was adapted for visualisation of the characteristic band pattern of HES hydrolysis products. The results indicate that the methods are efficient and useful for the screening of HES in urine.