Progressive multifocal leukoencephalopathy as an AIDS-defining condition in a patient with high CD4+ T-lymphocyte count

We present the case of a 31-year-old man with acute manifestation of progressive multifocal leukoencephalopathy (PML) as an AIDS-defining disease. The patient presented with a three-day history of neurological disease, brain lesions without mass effect or contrast uptake and a slightly increased protein concentration in cerebrospinal fluid. A serological test for HIV was positive and the CD4+ T-cell count was 427/mm³. Histological examination of the brain tissue revealed abnormalities compatible with PML. The disease progressed despite antiretroviral therapy, and the patient died three months later. PML remains an important cause of morbidity and mortality among HIV-infected patients.

Evaluation of CD4+CD25+ T lymphocyte response time kinetics in patients with chronic Chagas disease after in vitro stimulation with recombinant Trypanosoma cruzi antigens

Introduction CD4+CD25+ T lymphocytes have been implicated in the regulation of host inflammatory response against Trypanosoma cruzi, and may be involved in the clinical course of the disease. Methods Peripheral blood mononuclear cells from patients with chronic Chagas disease were cultured in the presence of T. cruzi recombinant antigens and assayed for lymphocytes at distinct time points. Results It was possible to differentiate clinical forms of chronic Chagas disease at days 3 and 5 according to presence of CD4+CD25+ T cells in cell cultures. Conclusions Longer periods of cell culture proved to be potentially valuable for prospective evaluations of CD4+CD25+ T lymphocytes in patients with chronic Chagas disease.

Nutritional status and CD4 cell counts in patients with HIV/AIDS receiving antiretroviral therapy

Introduction Even with current highly active antiretroviral therapy, individuals with AIDS continue to exhibit important nutritional deficits and reduced levels of albumin and hemoglobin, which may be directly related to their cluster of differentiation 4 (CD4) cell counts. The aim of this study was to characterize the nutritional status of individuals with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) and relate the findings to the albumin level, hemoglobin level and CD4 cell count. Methods Patients over 20 years of age with AIDS who were hospitalized in a university hospital and were receiving antiretroviral therapy were studied with regard to clinical, anthropometric, biochemical and sociodemographic characteristics. Body mass index, percentage of weight loss, arm circumference, triceps skinfold and arm muscle circumference were analyzed. Data on albumin, hemoglobin, hematocrit and CD4 cell count were obtained from patient charts. Statistical analysis was performed using Fisher's exact test, Student's t-test for independent variables and the Mann-Whitney U-test. The level of significance was set to 0.05 (α = 5%). Statistical analysis was performed using Statistical Package for the Social Sciences (SPSS) 17.0 software for Windows. Results Of the 50 patients evaluated, 70% were male. The prevalence of malnutrition was higher when the definition was based on arm circumference and triceps skinfold measurement. The concentrations of all biochemical variables were significantly lower among patients with a body mass index of less than 18.5kg/m2. The CD4 cell count, albumin, hemoglobin and hematocrit anthropometric measures were directly related to each other. Conclusions These findings underscore the importance of nutritional follow-up for underweight patients with AIDS, as nutritional status proved to be related to important biochemical alterations.

Implementação de metodologias para a contagem da flora específica do iogurte e de bifidobactérias na empresa SGS Portugal, S.A.

É do conhecimento geral que a população portuguesa está cada vez mais preocupada com a sua saúde e qualidade de vida, bem como em mudar alguns dos seus hábitos alimentares. Consequentemente, a procura por alimentos que proporcionam, para além de boas propriedades organolépticas e nutritivas, benefícios à saúde do consumidor, aumentou significativamente nas últimas décadas, onde os produtos lácteos fermentados surgem em grande destaque. Vários são os microrganismos empregues no desenvolvimento de produtos lácteos fermentados. Na produção de iogurtes, são utilizadas culturas lácteas tradicionais (Lactobacillus delbrueckii subsp. bulgaricus e Streptococcus thermophilus), também denominadas por flora específica do iogurte, e/ou bactérias probióticas, como as bifidobactérias, devido às suas boas características tecnológicas, terapêuticas e sensoriais. O trabalho descrito nesta dissertação de mestrado foca-se na validação e implementação de metodologias que permitem a contagem da flora específica do iogurte e de bifidobactérias, em iogurtes, na empresa SGS Portugal, S.A. A enumeração da flora específica do iogurte teve como base a norma ISO 7889:2003, por meio da técnica de sementeira por incorporação no meio de cultura apropriado e posterior contagem em placa dos microrganismos presentes. Foram analisadas 20 amostras de iogurtes, obtendo-se valores de contagens da flora específica do iogurte na ordem de 108 UFC/g de produto, indo ao encontro dos valores estabelecidos pela legislação portuguesa em vigor. A enumeração de bifidobactérias teve como base a norma ISO 29981:2010, por meio da técnica de sementeira por incorporação no meio de cultura apropriado e posterior contagem em placa dos microrganismos presentes. Das 20 amostras de iogurtes probióticos analisadas, apenas 15 obtiveram resultados positivos, apresentando valores de contagens de bifidobactérias superiores a 106 UFC/g de produto. Estas amostras, para além de apresentarem valores de acordo com os valores estabelecidos pela legislação portuguesa em vigor, são terapeuticamente eficazes, durante todo o prazo de validade do produto, uma vez que apresentam valores de contagem de bactérias probióticas superiores a 106 UFC/g, sendo este o padrão mínimo necessário para que um iogurte seja considerado como probiótico. Com base nos resultados obtidos e no cumprimento dos parâmetros preconizados nos procedimentos de validação de métodos microbiológicos, foi possível a validação destas metodologias e as suas implementações na empresa SGS Portugal, S.A.

Effector memory CD4+ T cells are associated with cognitive performance in a senior population

Objective: Immunosenescence and cognitive decline are common markers of the aging process. Taking into consideration the heterogeneity observed in aging processes and the recently described link between lymphocytes and cognition, we herein explored the possibility of an association between alterations in lymphocytic populations and cognitive performance. Methods: In a cohort of cognitively healthy adults (n = 114), previously characterized by diverse neurocognitive/psychological performance patterns, detailed peripheral blood immunophenotyping of both the innate and adaptive immune systems was performed by flow cytometry. Results: Better cognitive performance was associated with lower numbers of effector memory CD4(+) T cells and higher numbers of naive CD8(+) T cells and B cells. Furthermore, effector memory CD4(+) T cells were found to be predictors of general and executive function and memory, even when factors known to influence cognitive performance in older individuals (e.g., age, sex, education, and mood) were taken into account. Conclusions: This is the first study in humans associating specific phenotypes of the immune system with distinct cognitive performance in healthy aging.

Depressão em pacientes HIV positivos: a realidade de um hospital português

Aims: Depression is the most common psychiatric disorder among people infected with HIV. This study aims to characterize the Hospital of Joaquim Urbano population of HIV-infected patients’ profile regarding depressive symptoms and whether they correlate with the analytical parameters most frequently evaluated in the context of infection by this virus – HIV viral load, CD4+ count and CD4+ percentage. Methods: We conducted an observational descriptive and analytical study. The participants’ level of depressive symptoms was assessed with the Beck Depression Inventory. The medical and psychiatric history and the analytical values of viral load, CD4+ count and CD4+ percentage were obtained by consulting the participants’ clinical processes. Results: A prevalence of 65.5% in HIV-infected patients’ depressive symptoms was found, with a considerable high percentage of subjects presenting with severe symptoms (32.7%). No associations between the depressive symptoms’ levels and CD4+ count, CD4+ percentage or viral load were found. However, depressive symptoms were associated with substance abuse and education level. Conclusions: The high prevalence of depressive symptoms found in this study reinforces the importance of monitoring this type of symptoms in HIV-infected subjects. The fact that there have been no associations between depressive symptoms and the analytical parameters evaluated is in line with previous studies.

Determinantes prognósticos em pacientes com síndrome coronariana aguda sem elevação do segmento ST

FUNDAMENTO: Em pacientes com Síndromes Coronarianas Agudas (SCA) sem Supradesnivelamento do Segmento ST (SST), sugere-se que uma série de marcadores (células inflamatórias, hiperglicemia e função renal) é capaz de identificar indivíduos com maior risco para eventos cardiovasculares. OBJETIVO: Avaliar o impacto desses parâmetros laboratoriais em desfechos intra-hospitalares de pacientes com SCA sem SST. MÉTODOS: Foram avaliados prospectivamente 195 pacientes admitidos consecutivamente com SCA sem SST. Foram registrados dados clínicos, demográficos e laboratoriais ao longo do período de internação no hospital, em relação à ocorrência ou não de eventos combinados. RESULTADOS: A idade média foi de 67 ± 12 anos, e 52% eram homens. Na análise da área sob a curva ROC, somente a razão neutrófilo/linfócito (AUC: 70%, IC95%: 56%-82%, p = 0,006) e a creatinina (AUC: 62%, IC95%: 50%-80%, p = 0,03) discriminaram aqueles pacientes com SCA sem SST que apresentaram algum desfecho. Os pacientes que sofreram algum evento adverso durante a internação apresentaram menores contagens de linfócitos (1502 ± 731 / mm³ vs. 2020 ± 862 / mm³; p = 0,002), menores taxas de filtração glomerular (51 ± 27 mL/min vs. 77±34 mL/min; p < 0,001) e maiores níveis séricos de creatinina (2,1 ± 2,7 mg/dL vs. 1,1 ± 1,3 mg/dL; p = 0,047) do que aqueles que tiveram uma hospitalização sem intercorrências. A análise de regressão logística demonstrou que as variáveis que permaneceram como preditores independentes e significativos foram: taxa de filtração glomerular (OR: 1,03; IC95%: 1,00-1,13; p = 0,002), e contagem de linfócitos (OR: 1,02; IC95%: 1,01-1,04; p = 0,03). CONCLUSÃO: A avaliação da função renal e a contagem de linfócitos fornecem uma informação potencialmente útil para a estratificação prognóstica em doentes com SCA sem SST.

Microbiologia da carne moida: 1. contagem total de bactérias

Como primeira etapa de um programa de estudos microbiológicos da carne moida comercializada em Piracicaba, quarenta amostras de carne bovina, provenientes de dois tipos de estabelecimento de venda a varejo (açougue e supermercado), correspondendo a dois sistemas de distribuição de carne (um antigo e outro recente), foram analisados quanto ao número total de bactérias, segundo as recomendações da "American Public Health Association". As amostras foram obtidas semanalmente pela manhã e à tarde, durante aproximadamente dois meses. A incubação das placas de Petri foi feita a 21º C (72 horas) e a 32º (48 horas). Os resultados podem ser resumidos como segue: 1) Contagens totais elevadas foram, em geral, observadas, com diversas amostras na faixa de 10(7) a 10(9) bactérias/grama; as médias obtidas foram 6,9 x 10(7) (21ºC) e 2,5 x 10(7) (32ºC) bactérias por grama. 2) A incubação feita a 21ºC resultou, em geral, em contagens mais elevadas que as correspondentes à temperatura de incubação de 32ºC; todavia, a diferença observada não foi estatìsticamente significativa. 3) As contagens obtidas para as amostras correspondentes ao sistema antigo de distribuição de carne foram significativamente mais elevadas que as correspondentes ao sistema recente, o que se relacionou com contagens mais elevadas obtidas para as amostras coletadas pela manhã no estabelecimento menor.

Contagem total de bactérias e enumeração de coliformes em cortes de varejo de carne bovina e em equipamentos de supermercados

No presente trabalho foi determinada a contagem total de bactérias (incubação a 32°C e a 5°C) existentes na superfície de alguns cortes de varejo de carne bovina comercializada por tres supermercados de Piracicaba, SP, bem como da superfície de diversos tipos de equipamento utilizado nas salas de desossa desses estabelecimentos. Foi determinado também o número mais provável (NMP) de coliformes totais e, para um grupo de amostras de carne, o NMP de Escherichia coli. Para um dos supermercados (o mais antigo e menos adequado sob o aspecto sanitário) foi possível constatar uma diferença significativa entre as contagens totais em cortes recebidos já desossados pelo estabelecimento, a favor destes, e as contagens em cortes preparados na sala de desossa. O equipamento, tanto no estado considerado limpo (após o dia de trabalho) e aquele em uso apresentaram contagens totais excessivas; não houve diferença significativa entre um e outro caso, e, em ambos, a temperatura de incubação 32°C resultou em valores estatisticamente superiores aos obtidos com a incubação a 5°C. As contagens provenientes de análises de superfícies de madeira foram significativamente mais elevadas que as verificadas em superfícies metálicas (serra elétrica, facas, afiadores). A ocorrência de coliformes totais foi geral, tanto na carne como no equipamento, muitas vezes em números elevados. Em amostras de carne, analisadas quanto à presença e ao número de coliformes totais e de Escherichia coli, verificou-se que 96% daquelas positivas para coliformes totais também o eram para esta bactéria. Atras de material em uso foram positivas quanto à ocorrência de coliformes totais. Tal como ocorreu a carne, o número mais possível (NMP) assumiu, em vários casos, elevados valores. Embora não comprovado estatisticamente, os resultados sugerem uma maior ocorrência desse grupo de microrganismos nas superfícies de madeira (mesa e ou cepo).

The role of Notch in the differentiation of CD4⁺ T helper cells.

CD4⁺ T helper cells are playing critical roles in host defense to pathogens and in the maintenance of immune homeostasis. Naïve CD4⁺T cells, upon antigen-specific recognition, receive signals to differentiate into distinct effector T helper cell subsets characterized by their pattern of cytokine production and specific immune functions. A tight balance between these different subsets ensures proper control of the immune response. There is increasing evidence revealing an important role for Notch signaling in the regulation of CD4⁺T helper cell differentiation or function in the periphery. However, the exact mechanisms involved remain unclear and appear contradictory. In this review, we summarize current knowledge and discuss recent advances in the field to reconcile different views on the role of Notch signaling in the differentiation of functional T helper subsets.

Profiling of T-cell receptor signaling complex assembly in human CD4 T-lymphocytes using RP protein arrays.

The RP protein (RPP) array approach immobilizes minute amounts of cell lysates or tissue protein extracts as distinct microspots on NC-coated slide. Subsequent detection with specific antibodies allows multiplexed quantification of proteins and their modifications at a scale that is beyond what traditional techniques can achieve. Cellular functions are the result of the coordinated action of signaling proteins assembled in macromolecular complexes. These signaling complexes are highly dynamic structures that change their composition with time and space to adapt to cell environment. Their comprehensive analysis requires until now relatively large amounts of cells (>5 x 10(7)) due to their low abundance and breakdown during isolation procedure. In this study, we combined small scale affinity capture of the T-cell receptor (TCR) and RPP arrays to follow TCR signaling complex assembly in human ex vivo isolated CD4 T-cells. Using this strategy, we report specific recruitment of signaling components to the TCR complex upon T-cell activation in as few as 0.5 million of cells. Second- to fourth-order TCR interacting proteins were accurately quantified, making this strategy specially well-suited to the analysis of membrane-associated signaling complexes in limited amounts of cells or tissues, e.g., ex vivo isolated cells or clinical specimens.

Phenotypic heterogeneity of antigen-specific CD4 cells under different conditions of antigen persistence and antigen load

RESUME Nous n'avons pas de connaissance précise des facteurs à l'origine de l'hétérogénéité phénotypique des cellules T CD4 mémoires. Une troisième population phénotypique des cellules T CD4 mémoires, caractérisée par les marqueurs CD45RA+CCR7- a été identifiée dans cette étude. Cette population présente un état de différentiation avancée, comme en témoigne son histoire de réplication, ainsi que sa capacité de prolifération homéostatique. Les réponses des cellules T CD4 mémoires à différentes conditions de persistance et charge antigénique ont trois patterns phénotypiques différents, caractérisés par les marqueurs CD45RA et CCR7. La réponse CD4 mono -phénotypique CD45RA-CCR7+ ou CD45RA- CCR7- est associée à des conditions d'élimination de l'antigène (telle la réponse CD4 tétanos spécifique) ou à des conditions de persistance antigénique et de virémie élevée (telle la réponse HIV chronique ou la primo-infection CMV) respectivement. D'autre part, les réponses T CD4 multi -phénotypiques CD45RA-CCR7+ sont associées à des conditions d'exposition antigénique prolongée et de faible virémie (telles les infections CMV, EBV et HSV ou les infections HIV chez les long term non progressons). La réponse mono -phénotypique CD45RA- CCR7+ est propre aux cellules T CD4 secrétant de IL2, définies également comme centrales mémoires, la réponse CD45RA- CCR7- aux cellules T CD4 secrétant de l'IFNγ et finalement la réponse mufti-phénotypique aux cellules T CD4 secrétant à la fois de l'IL2 et de l' IFNγ. En conclusion, ces résultats témoignent d'une régulation de l'hétérogénéité phénotypique par l'exposition et la charge antigénique. ABSTRACT The factors responsible for the phenotypic heterogeneity of memory CD4 T cells are unclear. In the present study, we have identified a third population of memory CD4 T cells characterized as CD45RA+CCRT that, based on its replication history and the homeostatic proliferative capacity, was at an advanced stage of differentiation. Three different phenotypic patterns of memory CD4 T cell responses were delineated under different conditions of antigen (Ag) persistence and load using CD45RA and CCR7 as markers of memory T cells. Mono-phenotypic CD45RA'CCR7+ or CD45RA'CCR7' CD4 T cell responses were associated with conditions of Ag clearance (tetanus toxoid-specific CD4 T cell response) or Ag persistence and high load (chronic HIV-1 and primary CMV infections), respectively. Multi-phenotypic CD45RA CCR7+, CD45RA'CCRT and CD45RA+CCRT CD4 T cell responses were associated with protracted Ag exposure and low load (chronic CMV, EBV and HSV infections and HIV-1 infection in long-term nonprogressors). The mono-phenotypic CD45RA'CCR7+ response was typical of central memory (TCM) IL-2-secreting CD4 T cells, the mono-phenotypic CD45RA CCRT response of effector memory (TEM) IFN-γ -secreting CD4 T cells and the multi-phenotypic response of both IL-2- and IFN-γ -secreting cells. The present results indicate that the heterogeneity of different Ag-specific CD4 T cell responses is regulated by Ag exposure and Ag load.

Dynamics of HIV latency and reactivation in a primary CD4+ T cell model.

HIV latency is a major obstacle to curing infection. Current strategies to eradicate HIV aim at increasing transcription of the latent provirus. In the present study we observed that latently infected CD4+ T cells from HIV-infected individuals failed to produce viral particles upon ex vivo exposure to SAHA (vorinostat), despite effective inhibition of histone deacetylases. To identify steps that were not susceptible to the action of SAHA or other latency reverting agents, we used a primary CD4+ T cell model, joint host and viral RNA sequencing, and a viral-encoded reporter. This model served to investigate the characteristics of latently infected cells, the dynamics of HIV latency, and the process of reactivation induced by various stimuli. During latency, we observed persistence of viral transcripts but only limited viral translation. Similarly, the reactivating agents SAHA and disulfiram successfully increased viral transcription, but failed to effectively enhance viral translation, mirroring the ex vivo data. This study highlights the importance of post-transcriptional blocks as one mechanism leading to HIV latency that needs to be relieved in order to purge the viral reservoir.