Comparison of dengue infection in human mononuclear leukocytes with mosquito C6/36 and mammalian Vero cells using flow cytometry to detect virus antigen

Fluorescent activated cell sorter (FACS) analysis is useful for the detection of cellular surface antigens and intracellular proteins. We used this methodology in order to detect and quantify dengue antigens in highly susceptible cells such as clone C6/36 (Aedes albopictus) and Vero cells (green monkey kidney). Additionally, we analyzed the infection in vitro of human peripheral blood mononuclear leukocytes (PBML). FACS analysis turned out to be a reliable technique to quantify virus growth in traditional cell cultures of C6/36 as well as Vero cells. High rates of infection were achieved with a good statistical correlation between the virus amount used in infection and the percentage of dengue antigen containing cells detected in infected cultures. We also showed that human monocytes (CD14+) are preferred target cells for in vitro dengue infection among PBML. Monocytes were much less susceptible to virus infection than cell lines but they displayed dengue antigens detected by FACS five days after infection. In contrast, lymphocytes showed no differences in their profile for dengue specific immunofluorescence. Without an animal model to reproduce dengue disease, alternative assays have been sought to correlate viral virulence with clinical manifestations and disease severity. Study of in vitro interaction of virus and host cells may highlight this relationship.

Analyse de montres contrefaites possédant un bracelet parfumé: démarche de détection des composés volatils et vecteur d'information sur le phénomène

Comment l'analyse chimique des produits contrefaits peut-elle contribuer à mieux comprendre le phénomène de la contrefaçon?Pour appréhender cette question, une approche novatrice basée sur l'analyse par SPME GC-MS des composés volatils de trente-neuf bracelets parfumés présents sur des contrefaçons horlogères a été mise en oeuvre.La détection de plusieurs dizaines de composés par montre a permis de définir des profils chimiques discriminants. Au total, trois groupes de montres possédant des profils comparables ont été détectés. Ces groupes ont été mis en perspective de liens physiques détectés sur la base du marquage des bracelets (marques et logos) par la fédération de l'industrie horlogère suisse (FH) et des informations spatiotemporelles des saisies. Les montres provenant d'une même saisie présentent systématiquement des liens physiques, mais pas forcément des profils chimiques similaires. Il en ressort que les profils chimiques peuvent fournir une information complémentaire à l'analyse des marquages, qu'ils varient peu dans le temps et que des montres liées chimiquement sont retrouvées dans le monde entier. Cette étude exploratoire révèle ainsi le potentielle d'exploitation de techniques d'analyse complémentaires pour mieux comprendre les modes de production, voire de diffusion de la contrefaçon. Finalement, la méthode analytique a permis de détecter des composés de plastiques en sus des constituants de parfums. Ce résultat laisse entrevoir la possibilité d'exploiterla méthode pour une large gamme de produits contrefaits.

A Heminested Polymerase Chain Reaction for the Detection of Brazilian Rabies Isolates from Vampire Bats and Herbivores

A heminested-PCR (hn-PCR) using primers to the nucleoprotein-coding gene in a nested set was evaluated in the detection of Brazilian strains of rabies virus (RV). A representative number of RV nucleoprotein sequences belonging to genotype 1 were aligned. Based on such alignment, primers were directed to highly conserved regions. All 42 clinical samples positive by both fluorescent antibody and mouse inoculation tests were also positive by the hn-PCR. Brain tissue that had been left to decompose, obtained from an experimentally inoculated mouse was tested by hn-PCR and yielded positive results. In conclusion, primers designed here were capable of amplifying Brazilian RV isolates obtained from a rural epidemiological cycle.

Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation

Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance.

Investigating the genetic variation underlying episodicity in major depressive disorder: Suggestive evidence for a bipolar contribution.

BACKGROUND: Highly recurrent major depressive disorder (MDD) has reportedly increased risk of shifting to bipolar disorder; high recurrence frequency has, therefore, featured as evidence of 'soft bipolarity'. We aimed to investigate the genetic underpinnings of total depressive episode count in recurrent MDD. METHODS: Our primary sample included 1966 MDD cases with negative family history of bipolar disorder from the RADIANT studies. Total episode count was adjusted for gender, age, MDD duration, study and center before being tested for association with genotype in two separate genome-wide analyses (GWAS), in the full set and in a subset of 1364 cases with positive family history of MDD (FH+). We also calculated polygenic scores from the Psychiatric Genomics Consortium MDD and bipolar disorder studies. RESULTS: Episodicity (especially intermediate episode counts) was an independent index of MDD familial aggregation, replicating previous reports. The GWAS produced no genome-wide significant findings. The strongest signals were detected in the full set at MAGI1 (p=5.1×10(-7)), previously associated with bipolar disorder, and in the FH+ subset at STIM1 (p=3.9×10(-6) after imputation), a calcium channel signaling gene. However, these findings failed to replicate in an independent Munich cohort. In the full set polygenic profile analyses, MDD polygenes predicted episodicity better than bipolar polygenes; however, in the FH+ subset, both polygenic scores performed similarly. LIMITATIONS: Episode count was self-reported and, therefore, subject to recall bias. CONCLUSIONS: Our findings lend preliminary support to the hypothesis that highly recurrent MDD with FH+ is part of a 'soft bipolar spectrum' but await replication in larger cohorts.

PFAPA syndrome is not a sporadic disease.

OBJECTIVES: To determine whether PFAPA (periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis) patients have a positive family history (FH) for recurrent fever syndromes. METHOD: For all patients with PFAPA seen in two paediatric rheumatology centres (Romandy, Switzerland and Bordeaux, France), parents were interviewed to record the FH for periodic fever. As controls, we interviewed a group of children without history of recurrent fever. RESULTS: We recruited 84 patients with PFAPA and 47 healthy children. The FH for recurrent fever (without an infectious cause and recurring for at least half a year) was positive in 38/84 (45%), and was positive for PFAPA (diagnosis confirmed by a physician) in 10/84 (12%) of the PFAPA patients. For 29 of the 38 patients with positive FH, the affected person was a sibling or a parent. None of the healthy children had a positive FH for recurrent fever or PFAPA. A positive FH for rheumatological diseases was seen in both groups of children. CONCLUSION: These data show that a significant percentage of PFAPA patients present a positive FH of recurrent fever and PFAPA. This familial susceptibility suggests a potential genetic origin for this syndrome.

Capacity-approaching rate function for layered multiantenna architectures

The simultaneous use of multiple transmit and receive antennas can unleash very large capacity increases in rich multipath environments. Although such capacities can be approached by layered multi-antenna architectures with per-antenna rate control, the need for short-term feedback arises as a potential impediment, in particular as the number of antennas—and thus the number of rates to be controlled—increases. What we show, however, is that the need for short-term feedback in fact vanishes as the number of antennas and/or the diversity order increases. Specifically, the rate supported by each transmit antenna becomes deterministic and a sole function of the signal-to-noise, the ratio of transmit and receive antennas, and the decoding order, all of which are either fixed or slowly varying. More generally, we illustrate -through this specific derivation— the relevance of some established random CDMA results to the single-user multi-antenna problem.

Multiuser detection in a dynamic environment — Part II: Joint user identification and parameter estimation

The problem of jointly estimating the number, the identities, and the data of active users in a time-varying multiuser environment was examined in a companion paper (IEEE Trans. Information Theory, vol. 53, no. 9, September 2007), at whose core was the use of the theory of finite random sets on countable spaces. Here we extend that theory to encompass the more general problem of estimating unknown continuous parameters of the active-user signals. This problem is solved here by applying the theory of random finite sets constructed on hybrid spaces. We doso deriving Bayesian recursions that describe the evolution withtime of a posteriori densities of the unknown parameters and data.Unlike in the above cited paper, wherein one could evaluate theexact multiuser set posterior density, here the continuous-parameter Bayesian recursions do not admit closed-form expressions. To circumvent this difficulty, we develop numerical approximationsfor the receivers that are based on Sequential Monte Carlo (SMC)methods (“particle filtering”). Simulation results, referring to acode-divisin multiple-access (CDMA) system, are presented toillustrate the theory.

Follicular helper T cells: implications in neoplastic hematopathology.

A distinct subset of T helper cells, named follicular T helper cells (T(FH), has been recently described. T(FH) cells are characterized by their homing capacities in the germinal centers of B-cell follicles where they interact with B cells, supporting B-cell survival and antibody responses. T(FH) cells can be identified by the expression of several markers including the chemokine CXCL13, the costimulatory molecules PD1 and inducible costimulator, and the transcription factor BCL6. They appear to be relevant markers for the diagnosis of angioimmunoblastic T-cell lymphoma (AITL) and have helped to recognize subsets of peripheral T-cell lymphoma, not otherwise specified, with nodal or cutaneous presentation expressing T(FH) antigens that might be related to AITL. In B-cell neoplasms, T(FH) cells are present within the microenvironment of nodular lymphocyte-predominant Hodgkin lymphoma and follicular lymphoma, where they likely support the growth of neoplastic germinal center-derived B cells. Interestingly, the amount of PD1+ cells in the neoplastic follicles might have a favorable impact on the outcome of follicular lymphoma patients. Altogether, the availability of antibodies directed to T(FH)-associated molecules has important diagnostic and prognostic implications in hematopathology. In addition, T(FH) cells could represent interesting targets in T(FH)-derived lymphomas such as AITL, or in some B-cell neoplasms where they act as part of the tumor microenvironment.

Professional knowledge in primary health care of the person/family in mental distress: Le Boterf perspective

Aim of this study was to identify knowing-doing actions constituted the practice of Family Health (FH), in view of nurses in relation to the person and family care in mental distress in terms of professional knowledge of Le Boterf. Method: Descriptive exploratory qualitative study, to deepen contruction of nurse in FH. The survey was conducted in 3 Units FH. Result: Doing a thematic analysis, came to the following categories: “Knowing how to act and react with relevance”; “Knowing how to combine resources and mobilize them in a professional context”; “Knowing how to interact with multiple knowledges”; “Knowing how to transpose”; “Knowing how to learn and knowing how to learn to learn”; “Knowing how to engage”. Final considerations: the greatest difficulty was "be able to transpose," and that the daily demand of the FH teams requires a lot of this knowledge. Little transposition of knowing-doing in real situations has been verified.

Factores Humanos e Direito Internacional Aeronáutico

O objectivo deste trabalho, no âmbito do direito Internacional Aeronáutico, é analisar os problemas relacionados com o homem no sistema de aviação que constituem factores de risco para a segurança do mesmo sistema e a reacção dos Estados e organizações ligadas à aviação na criação de regulamentos, normas e procedimentos para evitar incidentes e acidentes aeronáuticos. Basicamente este trabalho vai ter em consideração a segurança e regulamentação. A segurança vai ser tratada nas suas vertentes Safety e Security na sua interacção com Factores Humanos (FH) e com o direito internacional aeronáutico. A legislação como medida preventiva ditada pela experiencias adquiridas com vários incidentes que sensibilizaram o mundo vai estar atenta na questão de factores humanos. As normas e regulamentos de Segurança Pública referentes à Aviação internacional vão aparecer na orientação de estruturas e legislação de cada Estado. Ressalta ainda percepção de fraqueza no combate das causas dos incidentes aeronáuticos apesar dos esforços dispensados pelos Estados e organizações da Aviação Internacional no sentido de contrariar as insuficiências de certas normas existentes. Ilumina a necessidade de se continuar a criar outras regras para melhorar o sistema da aviação em que o Homem é considerado o elemento central, mas fraco, que carece de atenção particularmente especial. Vários acidentes e em especial incidentes como o caso de 11 de Setembro, por serem tema de direito aeronáutico Internacional e de Segurança Pública, atingiram a consciência da comunidade da aviação e fizeram com que os Estados e comunidade aeronáutica em geral mudassem de atitude nas situações que perigam a aviação. Neste trabalho de Direito internacional aéreo são realçados alguns esforços já despendidos pelos Estados e pelas Organizações ligadas a aviação no sentido de se criar normas, regulamentos e procedimentos para uma aviação mais segura, com menos riscos de acidentes e incidentes. Os Estados, as Organizações Internacionais (IATA, IFALPA) e os Construtores de aviões ao reconhecerem a impossibilidade de eliminar os incidentes e acidentes uniram-se no esforço dos estudos de factores humanos para ajudar na criação de normas e desenvolver procedimentos que minimizam os riscos que podem por em causa a segurança na aviação. A Segurança Operacional (Safety), a Segurança contra os actos ilícitos (Security), os Factores Humanos e Direito Internacional Aeronáutico constituem palavras-chaves deste trabalho.

Spatio-temporal Brain Dynamics Mediating Post-error Behavioral Adjustments.

Optimal behavior relies on flexible adaptation to environmental requirements, notably based on the detection of errors. The impact of error detection on subsequent behavior typically manifests as a slowing down of RTs following errors. Precisely how errors impact the processing of subsequent stimuli and in turn shape behavior remains unresolved. To address these questions, we used an auditory spatial go/no-go task where continual feedback informed participants of whether they were too slow. We contrasted auditory-evoked potentials to left-lateralized go and right no-go stimuli as a function of performance on the preceding go stimuli, generating a 2 × 2 design with "preceding performance" (fast hit [FH], slow hit [SH]) and stimulus type (go, no-go) as within-subject factors. SH trials yielded SH trials on the following trials more often than did FHs, supporting our assumption that SHs engaged effects similar to errors. Electrophysiologically, auditory-evoked potentials modulated topographically as a function of preceding performance 80-110 msec poststimulus onset and then as a function of stimulus type at 110-140 msec, indicative of changes in the underlying brain networks. Source estimations revealed a stronger activity of prefrontal regions to stimuli after successful than error trials, followed by a stronger response of parietal areas to the no-go than go stimuli. We interpret these results in terms of a shift from a fast automatic to a slow controlled form of inhibitory control induced by the detection of errors, manifesting during low-level integration of task-relevant features of subsequent stimuli, which in turn influences response speed.

Frações da matéria orgânica do solo após três décadas de cultivo de eucalipto no Vale do Rio Doce-MG

As mudanças nas distintas frações da matéria orgânica do solo (MOS) sob cultivo do eucalipto são pouco conhecidas. Assim, o presente trabalho teve como objetivo avaliar o impacto do cultivo do eucalipto nos estoques de C das diversas frações da MOS, em solos ocupados anteriormente por pastagens degradadas, bem como verificar qual fração da MOS constitui-se um indicador mais sensível à mudança de uso do solo. O estudo foi realizado em plantações comerciais de eucalipto em dois locais na região do Vale do Rio Doce-MG (Belo Oriente e Virginópolis), onde foram determinados em amostras dos solos: C orgânico total (COT), C da fração leve (leve livre - FLL e leve oclusa - FLO), da fração pesada (areia - AR, silte + argila - S + A e argila - ARG), da biomassa microbiana (BM) e das frações húmicas (ácidos fúlvicos - FAF, ácidos húmicos - FAH e huminas - FH). Os resultados indicaram que, de maneira geral, os solos sob os diferentes tipos de uso em Virginópolis, em virtude da menor temperatura média anual e maior teor de argila, apresentaram maiores estoques de C em todas as frações da MOS em relação a Belo Oriente. Assim, o seqüestro de C no solo pelo cultivo do eucalipto foi maior em Virginópolis (14,2 t ha-1) que em Belo Oriente (10,0 t ha-1), resultando numa taxa de seqüestro de C de aproximadamente 0,42 e 0,29 t ha-1 ano-1 , respectivamente. Em Belo Oriente, o cultivo do eucalipto favoreceu o aumento no estoque de C das frações ARG, S + A e FH no solo. Comportamento semelhante foi observado para o C das FLL, FAF e FAH, em Virginópolis. Dentre todas as frações da MOS analisadas, a BM e a FLO foram indicadores menos sensíveis às alterações na MOS após três décadas da mudança de uso do solo. Com isso, o COT e a fração leve livre, ácidos fúlvicos, ácidos húmicos e humina foram mais eficientes nesse sentido.

Ácidos Húmicos em solo fertirrigado no Vale do São Francisco

A aplicação de ácidos orgânicos, principalmente húmicos (AH), associada à adubação mineral e orgânica pode levar a alterações nas propriedades dos AH do solo. No Vale do São Francisco, o uso de ácidos orgânicos vem despertando o interesse de produtores que usam sistema de irrigação. Entretanto, não são conhecidas as modificações que podem ocorrer nas características das substâncias húmicas do solo quando são aplicados ácidos orgânicos associados à fertilização mineral e estercos. Os objetivos deste trabalho foram avaliar e caracterizar, por métodos espectroscópicos (UV-Vis, Infravermelho, Fluorescência e RPE), as possíveis mudanças qualitativas no AH extraído de solo fertirrigado com duas fontes de fertilizantes (orgânica e mineral), associadas à aplicação de ácidos orgânicos comerciais na cultura da goiabeira. O experimento foi conduzido na Embrapa Semiárido, em Petrolina - PE. Os tratamentos utilizados foram: F - fertirrigação com fertilizantes minerais (ureia, fosfato monoamônio, cloreto de potássio); FE - F + aplicação de 20 dm³/planta de esterco; FH - F + fertirrigação com 30 dm³ ha-1 de Codahumus 20®; FEH - fertirrigação mineral + aplicação de 20 dm³/planta de esterco + fertirrigação com 30 dm³ ha-1 de Codahumus 20®, e R- tratamento testemunha, sem adubação. A relação E4/E6 dos AH variou de 4,8 a 5,4 para os tratamentos FEH e F. Os dados obtidos por meio da espectroscopia FTIR sugerem mistura de características aromático-alifáticas, grande quantidade de grupos carboxílicos e menor número de grupos nitrogenados para os AH dos tratamentos FEH, FH e FE. Os valores obtidos do índice A465 variaram de 1,04 a 1,74, referentes aos AH dos tratamentos FEH e FE. Verificou-se uma concentração de radicais livres orgânicos no AH do tratamento FEH (2,66 spins g-1 10(17)) três vezes maior do que o AH do tratamento F (0,95 spins g-1 10(17)). O esterco foi o responsável pelo maior de policondensação e grau de humificação do ácido húmico dos tratamentos FE e FHE.

Compartimentos de carbono em latossolo vermelho sob cultivo de eucalipto e fitofisionomias de cerrado

Os objetivos deste trabalho foram avaliar o impacto do cultivo do eucalipto, em diferentes idades, e de fitofisionomias de Cerrado sobre os estoques de C orgânico total e de seus compartimentos e verificar qual dessas frações constitui-se em indicador mais sensível às mudanças de uso do solo. O estudo foi realizado em Brasília, DF, em Latossolo Vermelho distroférrico sob plantio de eucalipto com três (PEUC-3), seis (PEUC-6) e sete (PEUC-7) anos de idade e sob Cerrado senso stricto, Cerrado denso e campo sujo. Nessas áreas, amostras de solo foram coletadas nas camadas de 0-10 e 10-20 cm, para determinação dos estoques de C orgânico total (COT), carbono da fração leve livre (FLL) e carbono das frações húmicas (ácidos fúlvicos - FAF, ácidos húmicos - FAH e huminas - FH). Em ambas as camadas de solo, observaram-se maiores estoques de COT e de C nos compartimentos, nas áreas sob Cerrado denso e Cerrado, comparativamente àquelas cultivadas com eucalipto; houve recuperação dos estoques de C em todos os compartimentos, com o aumento do tempo de cultivo do eucalipto. O C-FLL, seguido do C-FAF e C-FAH, foram considerados os compartimentos mais sensíveis às mudanças no uso do solo, uma vez que as reduções em estoque foram de maior amplitude do que as observadas no COT e C-FH. A análise multivariada, que considera todos os compartimentos conjuntamente, evidenciou maior similaridade entre o Cerrado denso e Cerrado, bem como entre o campo sujo, PEUC-3, PEUC-6 e PEUC7, nas camadas de 0-10 e 10-20 cm.