Application of maldi-tof mass spectrometry in clinical virology: a review.

MALDI-TOF mass spectrometry is a diagnostic tool of microbial identification and characterization based on the detection of the mass of molecules. In the majority of clinical laboratories, this technology is currently being used mainly for bacterial diagnosis, but several approaches in the field of virology have been investigated. The introduction of this technology in clinical virology will improve the diagnosis of infections produced by viruses but also the discovery of mutations and variants of these microorganisms as well as the detection of antiviral resistance. This review is focused on the main current applications of MALDI-TOF MS techniques in clinical virology showing the state of the art with respect to this exciting new technology.

Creatine, central nervous system and creatine deficiency syndromes

It was long thought that most of brain creatine was of peripheral origin. However, recentworks have demonstrated that creatine crosses blood-brain barrier only with poor efficiency, and thatCNS must ensure parts of its creatine needs by its own creatine synthesis pathway, thank to the brainexpression of AGAT and GAMT (creatine synthesis) and SLC6A8 (creatine transporter). This newunderstanding of creatine metabolism and transport in CNS allows a better comprehension of creatinedeficiency syndromes, which are due to deficiencies in AGAT, GAMT and SLC6A8 and mainly affectthe brain of patients who show severe neurodevelopmental delay and present neurological symptomsin early infancy.

EPI Update, April 14, 2006

Weekly newsletter for Center For Acute Disease Epidemiology of Iowa Department of Public Health.

Ammonia toxicity to the brain: effects on creatine metabolism and transport and protective roles of creatine.

Hyperammonemia can provoke irreversible damage to the developing brain, with the formation of cortical atrophy, ventricular enlargement, demyelination or gray and white matter hypodensities. Among the various pathogenic mechanisms involved, alterations in cerebral energy have been demonstrated. In particular, we could show that ammonia exposure generates a secondary deficiency in creatine in brain cells, by altering the brain expression and activity of the genes allowing creatine synthesis (AGAT and GAMT) and transport (SLC6A8). On the other hand, it is known that creatine administration can exert protective effects in various neurodegenerative processes. We could also show that creatine co-treatment under ammonia exposure can protect developing brain cells from some of the deleterious effects of ammonia, in particular axonal growth impairment. This article focuses on the effects of ammonia exposure on creatine metabolism and transport in developing brain cells, and on the potential neuroprotective properties of creatine in the brain exposed to ammonium.

Ammonium alters creatine transport and synthesis in a 3D culture of developing brain cells, resulting in secondary cerebral creatine deficiency.

Hyperammonemic disorders in pediatric patients lead to poorly understood irreversible effects on the developing brain that may be life-threatening. We showed previously that some of these NH4+-induced irreversible effects might be due to impairment of axonal growth that can be protected under ammonium exposure by creatine co-treatment. The aim of the present work was thus to analyse how the genes of arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT), allowing creatine synthesis, as well as of the creatine transporter SLC6A8, allowing creatine uptake into cells, are regulated in rat brain cells under NH4+ exposure. Reaggregated brain cell three-dimensional cultures exposed to NH4Cl were used as an experimental model of hyperammonemia in the developing central nervous system (CNS). We show here that NH4+ exposure differentially alters AGAT, GAMT and SLC6A8 regulation, in terms of both gene expression and protein activity, in a cell type-specific manner. In particular, we demonstrate that NH4+ exposure decreases both creatine and its synthesis intermediate, guanidinoacetate, in brain cells, probably through the inhibition of AGAT enzymatic activity. Our work also suggests that oligodendrocytes are major actors in the brain in terms of creatine synthesis, trafficking and uptake, which might be affected by hyperammonemia. Finally, we show that NH4+ exposure induces SLC6A8 in astrocytes. This suggests that hyperammonemia increases blood-brain barrier permeability for creatine. This is normally limited due to the absence of SLC6A8 from the astrocyte feet lining microcapillary endothelial cells, and thus creatine supplementation may protect the developing CNS of hyperammonemic patients.

Vastaus ed. V. Annalan ym. kysymykseen koskeva Keski-Euroopan maista saapuneiden pakolaisten lukumäärä

Puhe

Comportamentos de saúde e doença numa comunidade Cabo-Verdiana em Lisboa

O objetivo foi compreender como os indivíduos se comportam em termos de saúde e o que fazem em caso de doença. Análise comparativa realçou semelhanças e divergências de práticas de saúde ou em caso de doença. O estudo foi efetuado numa amostra de 40 cabo-verdianos da primeira geração residentes na região de Lisboa, dividida em subgrupos: grupo social, geração e genero. Baseou-se em metodologia qualitativa com entrevistas semi-estruturadas. As práticas analisadas foram agrupadas em preventivas e de saúde, práticas utilizadas em episódios de doença, recursos para prevenção e tratamento, utilização de remédios caseiros e outros recursos ou terapeutas. Indivíduos experimentaram, ao nível das práticas, três sistemas de saúde que coexistiam em Cabo Verde, oficial, popular e tradicional e o recurso à religião. O discurso acerca das práticas de saúde e de doença demonstrou existirem diferenças, em alguns aspectos, entre grupos sociais e entre generos e gerações. Práticas de saúde destes imigrantes são idênticas às dos portugueses em contextos socioeconomicos semelhantes. Resultados sugerem existência de diferenças entre grupos sociais relativamente às práticas, na esfera da saúde e da doença. Mais que cultura e etnicidade, que se moldam às condições materiais de existência, neste estudo, o nível socioeconomico determinou as maiores diferenças a interferir nas práticas de saúde e doença, de grupo com cultura de base comum. Em geral, os indivíduos sobrevalorizaram sua identidade étnica e cultura de origem. Pertencimento a grupos sociais diferentes dá origem a partilha do sentimento de pertença cultural, mas não a comportamentos e práticas idênticos.

Creatine and guanidinoacetate transport at blood-brain and blood-cerebrospinal fluid barriers.

While it was thought that most of cerebral creatine is of peripheral origin, AGAT and GAMT are well expressed in CNS where brain cells synthesize creatine. While the creatine transporter SLC6A8 is expressed by microcapillary endothelial cells (MCEC) at blood-brain barrier (BBB), it is absent from their surrounding astrocytes. This raised the concept that BBB has a limited permeability for peripheral creatine, and that the brain supplies a part of its creatine by endogenous synthesis. This review brings together the latest data on creatine and guanidinoacetate transport through BBB and blood-CSF barrier (BCSFB) with the clinical evidence of AGAT-, GAMT- and SLC6A8-deficient patients, in order to delineate a clearer view on the roles of BBB and BCSFB in the transport of creatine and guanidinoacetate between periphery and CNS, and on brain synthesis and transport of creatine. It shows that in physiological conditions, creatine is taken up by CNS from periphery through SLC6A8 at BBB, but in limited amounts, and that CNS also needs its own creatine synthesis. No uptake of guanidinoacetate from periphery occurs at BBB except under GAMT deficiency, but a net exit of guanidinoacetate seems to occur from CSF to blood at BCSFB, predominantly through the taurine transporter TauT.

Creatine deficiency syndromes and the importance of creatine synthesis in the brain.

Creatine deficiency syndromes, due to deficiencies in AGAT, GAMT (creatine synthesis pathway) or SLC6A8 (creatine transporter), lead to complete absence or very strong decrease of creatine in CNS as measured by magnetic resonance spectroscopy. Brain is the main organ affected in creatine-deficient patients, who show severe neurodevelopmental delay and present neurological symptoms in early infancy. AGAT- and GAMT-deficient patients can be treated by oral creatine supplementation which improves their neurological status, while this treatment is inefficient on SLC6A8-deficient patients. While it has long been thought that most, if not all, of brain creatine was of peripheral origin, the past years have brought evidence that creatine can cross blood-brain barrier, however, only with poor efficiency, and that CNS must ensure parts of its creatine needs by its own endogenous synthesis. Moreover, we showed very recently that in many brain structures, including cortex and basal ganglia, AGAT and GAMT, while found in every brain cell types, are not co-expressed but are rather expressed in a dissociated way. This suggests that to allow creatine synthesis in these structures, guanidinoacetate must be transported from AGAT- to GAMT-expressing cells, most probably through SLC6A8. This new understanding of creatine metabolism and transport in CNS will not only allow a better comprehension of brain consequences of creatine deficiency syndromes, but will also contribute to better decipher creatine roles in CNS, not only in energy as ATP regeneration and buffering, but also in its recently suggested functions as neurotransmitter or osmolyte.

Novas cultivares de acerola (Malpighia emarginata DC): UEL 3 -- Dominga, UEL 4 -- Lígia e UEL 5 -- Natália

O programa de melhoramento genético de acerola, na Universidade Estadual de Londrina, iniciou-se em 1992, com a implatação de um pomar clonal constituído de genótipos selecionados no Norte do Paraná. As cultivares UEL 3-Dominga , UEL 4-Lígia, UEL 5-Natália são resultados de 5 ciclos de avaliações, onde foram considerados: precocidade, produtividade, conteúdo de vitamina C nos frutos, tolerância a pragas e doenças, tamanho e aparência dos frutos. As cultivares UEL 3-Dominga e UEL 5- Natália, destacam-se pela produtividade das plantas, conteúdo de vitamina C e pelo tamanho do fruto. Em Londrina, a frutificação dessas cultivares concentra-se nos meses de novembro a março. A cultivar UEL 4-Lígia destaca-se pela precocidade e produtividade que, em Londrina, se concentra nos meses de outubro a março. A cultivar UEL 3-Dominga apresenta peso médio de frutos igual a 6,31g, Vitamina C igual a 2906 e 1250 mg/100g em frutos verdes e maduros, respectivamente, acidez igual a 694,34 mg/100g e Brix igual a 9,2º. A cultivar UEL 4-Lígia apresenta peso médio dos frutos igual a 7,85g, vitamina C igual a 3579 e 1458 mg/100g em frutos verdes e maduros, respectivamente, acidez igual a 1110 mg/100g e Brix igual a 7,85º. A cultivar UEL 5-Natália apresenta peso médio dos frutos igual a 7,47g, Vitamina C igual a 3134,5 e 1098 mg/100g em frutos verdes e maduros, respectivamente, acidez igual a 725 mg/100g e Brix igual a 7,6º. Estas três cultivares são indicadas para plantio no Norte do Estado do Paraná.

Creatine deficiency syndomes : a new model of partial GAMT deficiency affecting brain cell development

Les syndromes de déficiences cérébrales en créatine (CCDS) sont dus à des mutations dans les gènes GATM et G AMT (codant pour les enzymes AGAT et G AMT de la voie de synthèse de créatine) ainsi que SLC6A8 (transporteur de créatine), et génèrent une absence ou une très forte baisse de créatine (Cr) dans le cerveau, mesurée par spectroscopic de résonance magnétique. Les patients CCDS développent des handicaps neurologiques sévères. Les patients AGAT et GAMT peuvent être traités avec des doses importantes de Cr, mais gardent dans la plupart des cas des séquelles neurologiques irréversibles. Aucun traitement efficace n'existe à ce jour pour la déficience en SLC6A8. Bien que de nombreux modèles aient été développés pour comprendre la Cr cérébrale en conditions physiologiques, les pathomécanismes des CCDS ne sont pas encore compris. Des souris transgéniques pour les gènes Gatm, Gamt et Slc6a8 ont été générées, mais elles ne miment que partiellement la pathologie humaine. Parmi les CCDS, la déficience en GAMT est la plus sévère, en raison de l'accumulation cérébrale de l'intermédiaire guanidinoacétate (GAA). Alors que la toxicité cérébrale du GAA a été étudiée par exposition directe au GAA d'animaux adultes sains, les mécanismes de la toxicité du GAA en condition de déficience en GAMT dans le cerveau en développement sont encore inconnus. Le but de ce projet était donc de développer un modèle de déficience en GAMT dans des cultures 3D primaires de cellules nerveuses de rat en agrégats par knock-down du gène GAMT, en utilisant un virus adéno-associé (AAV) induisant le mécanisme d'interférence à l'ARN (RNAi). Le virus scAAV2, à la multiplicité d'infection de 1000, s'est révélé le plus efficace pour transduire tous les types de cellules nerveuses des cultures (neurones, astrocytes, oligodendrocytes), et générer un knock-down maximal de la protéine GAMT de 85% (jour in vitro 18). Cette déficience partielle en GAMT s'est révélée insuffisante pour générer une déficience en Cr, mais a causé l'accumulation attendue de GAA, à des doses comparables aux niveaux observés dans le LCR des patients GAMT. Le GAA a induit une croissance axonale anarchique accompagnée d'une baisse de l'apoptose naturelle, suivis par une induction tardive de mort cellulaire non-apoptotique. Le co-traitement par la Cr a prévenu tous les effets toxiques du GAA. Ce travail montre que l'accumulation de GAA en absence de déficience en Cr est suffisante pour affecter le développement du tissu nerveux, et suggère que des formes de déficiences en GAMT supplémentaires, ne présentant pas de déficiences en Cr, pourraient être découvertes par mesure du GAA, en particulier à travers les programmes récemment proposés de dépistage néonatal de la déficience en GAMT. -- Cerebral creatine deficiency syndromes (CCDS) are caused by mutations in the genes GATM and GAMT (respectively coding for the two enzymes of the creatine synthetic pathway, AGAT and GAMT) as well as SLC6A8 (creatine transporter), and lead to the absence or very strong decrease of creatine (Cr) in the brain when measured by magnetic resonance spectroscopy. Affected patients show severe neurological impairments. While AGAT and GAMT deficient patients can be treated with high dosages of Cr, most remain with irreversible brain sequelae. No treatment has been successful so far for SLC6A8 deficiency. While many models have helped understanding the cerebral Cr pathways in physiological conditions, the pathomechanisms underlying CCDS are yet to be elucidated. Transgenic mice carrying mutations in the Gatm, Gamt and Slc6a8 genes have been developed, but only partially mimic the human pathology. Among CCDS, GAMT deficiency is the most severe, due to the CNS accumulation of the guanidinoacetate (GAA) intermediate. While brain toxicity of GAA has been explored through direct GAA exposure of adult healthy animals, the mechanisms underlying GAA toxicity in GAMT deficiency conditions on the developing CNS are yet unknown. The aim of this project was thus to develop and characterize a GAMT deficiency model in developing brain cells by gene knockdown, by adeno-associated virus (AAV)-driven RNA interference (RNAi) in rat 3D organotypic primary brain cell cultures in aggregates. scAAV2 with a multiplicity of infection of 1000 was shown as the most efficient serotype, was able to transduce all brain cell types (neurons, astrocytes, oligodendrocytes) and to induce a maximal GAMT protein knockdown of 85% (day in vitro 18). Metabolite analysis showed that partial GAMT knockdown was insufficient to induce Cr deficiency but generated the awaited GAA accumulation at concentrations comparable to the levels observed in cerebrospinal fluid of GAMT-deficient patients. Accumulated GAA induced axonal hypersprouting paralleled with inhibition of natural apoptosis, followed by a later induction in non-apoptotic cell death. Cr supplementation led to the prevention of all GAA-induced toxic effects. This work shows that GAA accumulation without Cr deficiency is sufficient to affect CNS development, and suggests that additional partial GAMT deficiencies, which may not show the classical brain Cr deficiency, may be discovered through GAA measurement including by recently proposed neonatal screening programs for GAMT deficiency.

Cultural differences in web-based health information seeking of Finnish and Japanese students

Abstrakti

Caracterização morfométrica da bacia hidrográfica do rio Debossan, Nova Friburgo, RJ

O objetivo deste trabalho foi fazer a caracterização morfométrica a partir de alguns parâmetros físicos da bacia hidrográfica do rio Debossan, Nova Friburgo, RJ. Para isso, gerou-se inicialmente o Modelo Digital de Elevação Hidrologicamente Consistente (MDEHC) a partir de cartas topográficas do IBGE, na escala 1:50.000, utilizando o sistema de informações geográficas, através dos softwares ArcVIEW e Arc/INFO. A partir do MDEHC, foram calculados alguns parâmetros morfométricos para o estudo do comportamento hidrológico da bacia. A área de drenagem encontrada foi de 9,9156 km² e o perímetro, de 17,684 km. A bacia hidrográfica do rio Debossan tem formato alongado, coeficiente de compacidade de 1,5842, fator de forma de 0,3285 e índice de circularidade de 0,3985. A densidade de drenagem obtida para a bacia foi de 2,3579 km/km². A forma mais alongada da bacia hidrográfica indica que a precipitação pluviométrica sobre ela se concentra em diferentes pontos, concorrendo para amenizar a influência da intensidade de chuvas, as quais poderiam causar maiores variações da vazão do curso d'água.

L'Univers

1877/07/26 (Numéro 3579).