Patologia Clínica
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- Bactérias Multirresistentes Associadas aos Cuidados de Saúde num Hospital Pediátrico: Experiência de Cinco Anos Cuidados de Saúde num Hospital Pediátrico: Experiência de Cinco AnosPublication . Mação, P; Lopes, JC; Oliveira, H; Oliveira, G; Rodrigues, FINTRODUCTION: In recent years there has been an increase of infections caused by multidrug-resistant bacteria. Paediatric data are scarce, particularly at national level. AIMS: To analyse trends of health-care associated multidrug-resistant bacteria infections in a paediatric hospital. MATERIAL AND METHODS: A retrospective incidence study was conducted in medical, surgical and intensive care wards in a level III paediatric hospital, from January 2005 to December 2009. The studied multidrug-resistant bacteria were methicillin-resistant Staphylococcus aureus (MRSA), ESBL-producing bacilli, vancomycin-resistant Enterococcus spp, multidrug-resistant Pseudomonas aeruginosa (PA-MDR) and Acinetobacter baumannii resistant to carbapenems. Demographic, clinical and laboratory data, treatment and presence of risk factors for these infections were analysed. RESULTS: During the study period 106 multidrug-resistant bacteria were identified, related to 72 children, predominantly male (65.3%). The most frequently identified multidrug-resistant bacteria were MRSA (35.8%), PA-MDR (29.2%) and ESBL-producing bacilli (17.9%). Of the 106 multidrug-resistant bacteria, 45 (42.5%) were causing infection. During the study period the annual proportion of infections caused by multidrug-resistant bacteria went up from 32.0% in 2006 to 55.6% in 2009 (p = 0.376). The overall incidence rate of infection was 0.400 per 1 000 hospitalization-days, corresponding to 0.274 infections per 100 admissions, remaining stable over the five years. The most frequent infections were bloodstream (31.1%), intra-abdominal (20.0%), catheter-related (17.8%) and skin and soft tissue (11.1%). All children had risk factors and the most frequently identified were previous antibiotic therapy and underlying chronic disease (> 90%). Six children (13.3%) died during hospitalisation. CONCLUSIONS: During the study period, there was an increase in the proportion of multidrug-resistant bacteria but with no statistical significance. The overall incidence rate of multidrug-resistant bacteria infection remained stable. MRSA were the most frequently identified bacteria, followed by PA-MDR and ESBL-producing bacilli. Bloodstream infections were the most frequent infections, followed by intra-abdominal and catheter-related. All children had risk factors, mainly previous antibiotic therapy and chronic disease.
- Efeito do exercício físico no metabolismo lipídico dos idososPublication . Veríssimo, MT; Aragão, A; Sousa, A; Barbosa, B; Ribeiro, H; Costa, D; Saldanha, MHPURPOSE: To assess the effect of regular physical exercise on plasma lipids and apoproteins in an elderly group. MATERIAL AND METHODS: Sixty-three old people of both sexes, aged between 65 and 94, were randomly distributed between a test group (n = 31) and a control group (n = 32). The test group followed a program of physical exercise for eight months, with intensity of 60% to 80% of heart rate reserve, consisting of three weekly sessions, on alternate days, of 60 minutes each. Before the beginning of the program and eight months afterwards, blood samples were collected to assess total cholesterol, triglycerides, HDL-C, HDL2-C. HDL3-C, apo A1, apo B-100 and apo (a). The ratios total cholesterol/HDL-C, LDL-C/HDL-C, HDL2-C/HDL3-C and apo A1/apo B-100 were also calculated. RESULTS: In the test group total cholesterol, triglycerides, LDL-C, apo B-100, total cholesterol/HDL-C and LDL-C/HDL-C decreased significantly; HDL-C, HDL2-C, apo A1, HDL2-C/HDL3-C and apo A1/apo B-100 increased significantly; HDL3-C and apo (a) did not change. The control group did not present any significant alterations in any of the parameters assessed. CONCLUSIONS: Regular physical exercise decreases vascular risk and may help to reduce cardiovascular events in the elderly.
- Harmonemia: a universal strategy for flow cytometry immunophenotyping-A European LeukemiaNet WP10 studyPublication . Lacombe, F; Bernal, E; Bloxham, D; Couzens, S; Porta, M G D; Johansson, U; Kern, W; Macey, M; Matthes, T; Morilla, R; Paiva, A; Palacio, C; Preijers, F; Ratei, R; Siitonen, S; Allou, K; Porwit, A; Béné, M C
- Lifelong training improves anti-inflammatory environment and maintains the number of regulatory T cells in masters athletesPublication . Minuzzi, LG; Rama, L; Bishop, NC; Rosado, F; Martinho, A; Paiva, A; Teixeira, AMPURPOSE: The purpose of this study was to quantify and characterize peripheral blood regulatory T cells (Tregs), as well as the IL-10 plasma concentration, in Masters athletes at rest and after an acute exhaustive exercise test. METHODS: Eighteen Masters athletes (self-reported training: 24.6 ± 1.83 years; 10.27 ± 0.24 months and 5.45 ± 0.42 h/week per each month trained) and an age-matched control group of ten subjects (that never took part in regular physical training) volunteered for this study. All subjects performed an incremental test to exhaustion on a cycle ergometer. Blood samples were obtained before (Pre), 10 min into recovery (Post), and 1 h after the test (1 h). RESULTS: Absolute numbers of Tregs were similar in both groups at rest. Acute exercise induced a significant increase in absolute numbers of Tregs at Post (0.049 ± 0.021 to 0.056 ± 0.024 × 109/L, P = 0.029 for Masters; 0.048 ± 0.017 to 0.058 ± 0.020 × 109/L, P = 0.037 for control) in both groups. Treg mRNA expression for FoxP3, IL-10, and TGF-β in sorted Tregs was similar throughout the trials in both groups. Masters athletes showed a higher percentage of subjects expressing the FoxP3 (100% for Masters vs. 78% for Controls, P = 0.038) and TGF-β (89% for Masters vs. 56% for Controls, P = 0.002) after exercise and a higher plasma IL-10 concentration (15.390 ± 7.032 for Masters vs. 2.411 ± 1.117 for control P = 0.001, ES = 2.57) at all timepoints. KLRG1 expression in Tregs was unchanged. CONCLUSION: Our findings showed that Masters athletes have elevated anti-inflammatory markers and maintain the number of Tregs, and may be an adaptive response to lifelong training.
- Oxidative Stress, DNA, Cell Cycle/Cell Cycle Associated Proteins and Multidrug Resistance Proteins: Targets of Human Amniotic Membrane in Hepatocellular CarcinomaPublication . Mamede, AC; Guerra, S; Laranjo, M; Santos, K; Carvalho, MJ; Carvalheiro, T; Moura, P; Paiva, A; Abrantes, AM; Maia, CJ; Botelho, MFThe anticancer effects of human amniotic membrane (hAM) have been studied over the last decade. However, the action mechanisms responsible for these effects are not fully understood until now. Previously results reported by our team proved that hAM is able to induce cytotoxicity and cell death in hepatocellular carcinoma (HCC), a worldwide high incident and mortal cancer. Therefore, this experimental study aimed to investigate the cellular targets of hAM protein extracts (hAMPE) in HCC through in vitro studies. Our results showed that hAMPE is able to modify oxidative stress environment in all HCC cell lines, as well as its cell cycle. hAMPE differently targets deoxyribonucleic acid (DNA), P21, P53, β-catenin and multidrug resistance (MDR) proteins in HCC cell lines. In conclusion, hAMPE has several targets in HCC, being clear that the success of this treatment depends of a personalized therapy based on the biological and genetic characteristics of the tumor.
- Padrão clínico e laboratorial de sensibilização a fungosPublication . Santos, A; Carrapatoso, I; Rodrigues, F; Geraldes, L; Loureiro, C; Chieira, CObjectivo: Caracterizar clinicamente um grupo de doentes sensibilizados a fungos; comparar os resultados de diferentes testes diagnósticos; e definir um padrão de reactividade a espécies fúngicas comuns numa população de doentes com alergia respiratória. Métodos: Entre os doentes que consecutivamente realizaram testes cutâneos (TC) com bateria de fungos entre 2001 e 2005 (n=304), seleccionaram -se vinte com maior reactividade cutânea a pelo menos um dos seguintes fungos: Alternaria alternata, Aspergillus fumigatus, Candida albicans, Cladosporium herbarum e Penicillium chrysogenum. Para todos os doentes, analisaram -se os processos clínicos e realizaram -se TC, doseamento de IgE especifica (IgE) e immunoblotting (IB) aos cinco fungos mencionados. A concordância dos exames referidos foi avaliada através do coeficiente kappa (κ) de Cohen. Resultados: No grupo de 20 doentes, com idades compreendidas entre os 20 e os 60 anos, 90% apresentavam rinite (55% persistente (P) moderada -grave, 39% P ligeira e 6% intermitente ligeira), 80% asma (56% P moderada, 19% P ligeira e 25% intermitente), 35% sinusite, 5% polipose nasal e 35% conjuntivite. A maioria estava sensibilizada a mais do que um fungo e a outros aeroalergénios. Comparando os resultados dos exames realizados, observou-se uma concordância entre IgE e IB (κ=0,240; p=0,009), mas não entre TC e IgE (κ=0,09; p=0,303) ou TC e IB (κ=0,036; p=0,719). No estudo de IB, foram identificadas proteínas com diferentes pesos moleculares, alguns correspondendo a alergénios já descritos. Conclusões: Este estudo vem reforçar a discordância entre testes in vivo e in vitro na sensibilização a fungos. Mais estudos e melhores extractos alergénicos são necessários. Por enquanto, deve manter-se um elevado índice de suspeição, bem como uma combinação de exames complementares no diagnóstico de alergia a fungos.
- Padrões clínicos de hipersensibilidade a frutos: estudos in vivo e in vitroPublication . Carrapatoso, I; Faria, E; Rodrigues, F; Todo-Bom, A; Loureiro, C; Chieira, C
- Padrões clínicos e laboratoriais na hipersensibilidade ao camarão e reactividade cruzada com Dermatophagoides pteronyssinusPublication . Carrapatoso, I; Rodrigues, F; Geraldes, L; Faria, E; Todo-Bom, A; Loureiro, C; Chieira, CObjectivos: Investigar padrões clínicos em doentes com hipersensibilidade a camarão e a ocorrência de reactividade cruzada a Dermatophagoides pteronyssinus (Dp). Caracterizar as principais proteínas envolvidas na reactividade a camarão, em estudos de immunoblotting. Métodos: Avaliaram-se 20 doentes com clínica de hipersensibilidade imediata, após ingestão de camarão. Aplicou-se um questionário estandardizado, foram efectuados testes cutâneos por prick a alimentos e aeroalergénios e procedeu-se à determinação de IgE específica a camarão e Dp por UniCap e immunoblotting. A existência de eventual reactividade cruzada imunológica entre camarão e Dp foi investigada por estudos de inibição de immunoblotting. Resultados: A média de idades dos doentes era de 33±16 anos, sendo 13 do sexo masculino. As manifestações clínicas após ingestão de camarão foram urticária e/ou angioedema generalizados em 8 doentes, síndrome de alergia oral em 9 e anafilaxia em 3. Para além do camarão, foi detectada em todos os doentes sensibilização a outros crustáceos com sintomas semelhantes. Outros alimentos reportados em que se confirmou sensibilização foram moluscos -12 doentes e ovo -1. Em todos os doentes foi detectada sensibilização a Dp e sensibilização associada a barata em 12, a pólenes em 13, a cão em 5 e a gato em 5. Doseamentos de IgE específica a Dp e camarão superiores ou iguais a 3,5 KU/l foram encontrados, respectivamente, em 15 e 12 doentes. Considerando os estudos de immunoblotting a camarão, observou-se uma maior intensidade de ligação a bandas com peso molecular (PM) de 35 -40 KDa em 14 doentes, 17 KDa em 2, 22 KDa em um e 43-50 KDa nos restantes 3. Os estudos de inibição de immunoblotting evidenciaram reactividade cruzada em 15 doentes, recíproca camarão - Dp em 10 e não recíproca em 5, não sendo possível demonstrar a sua existência em 5. Conclusões: O PM encontrado para as bandas de maior intensidade nos estudos de immunoblotting a camarão sugere o envolvimento de tropomiosinas na maioria dos doentes, reforçando o papel destes panalergénios como alergénios major na hipersensibilidade a camarão em doentes sensibilizados a ácaros. Sensibilização simultânea a camarão e Dp pode ocorrer na ausência de reactividade cruzada e diferentes padrões de sensibilização deverão ser considerados.
- Padrões de sensibilização na hipersensibilidade a leguminosas. Estudo numa população da região Centro.Publication . Geraldes, L; Carrapatoso, I; Santos, A; Rodrigues, F; Todo-Bom, A; Faria, E; Chieira, CBackground: Foods from the legume family are used in the Mediterranean diet as a source of protein. In some cases, their ingestion triggers potentially life -threatening reactions. A high degree of immunological cross-reactivity has been described, but its clinical relevance is controversial. Aim: To carry out a clinical and laboratory characterisation of patients sensitised to leguminosae who had symptoms after the ingestion of at least one food from the leguminosae botanical family. Methods: We selected 13 consecutive patients from a food allergy outpatient clinic with allergy symptoms after the ingestion of leguminosae during the year of 2006. Case history data, skin prick tests (SPT) with commercial extracts of white bean, green bean, pea, chickpea, soya bean and peanut, and prick-to-prick (PP) tests to these legumes, both raw and boiled, and also to lupini bean and black -eyed bean were performed. Serum specific IgE and immunoblotting assays were also carried out. Results: Most patients had had symptoms with more than one legume (mean 3 legumes) with bean the most reported. The first culprit leguminosae reported was chickpea in 4 patients, bean in 4 patients and lupini bean in 3. Sensitisation to other legumes without clinical symptoms also occurred (mean 2 legumes). Anaphylaxis was described in 3 patients, to bean, soya and lupini bean, respectively. Asthma was connected to chickpea (2 patients) and to bean (1). One or more of the following symptoms: urticaria, angiooedema and oral allergy syndrome were reported by other patients. Symptomatic patients always had positive PP tests to leguminosae and most SPT were also positive. Conclusions: Lifethreatening reactions to leguminosae were found. PP tests used alone to raw and boiled legumes seem to be the most reliable diagnostic tests as in the case of other vegetable foods. However, immunoblotting assays are still important in the identification of new allergens.
- Phenotypic profile of expanded NK cells in chronic lymphoproliferative disorders: a surrogate marker for NK-cell clonalityPublication . Bárcena, P; Jara-Acevedo, M; Tabernero, MD; López, A; Sánchez, ML; García-Montero, AC; Muñoz-García, N; Vidriales, MB; Paiva, A; Lecrevisse, Q; Lima, M; Langerak, AW; Böttcher, S; van Dongen, JM; Orfão, A; Almeida, JCurrently, the lack of a universal and specific marker of clonality hampers the diagnosis and classification of chronic expansions of natural killer (NK) cells. Here we investigated the utility of flow cytometric detection of aberrant/altered NK-cell phenotypes as a surrogate marker for clonality, in the diagnostic work-up of chronic lymphoproliferative disorders of NK cells (CLPD-NK). For this purpose, a large panel of markers was evaluated by multiparametric flow cytometry on peripheral blood (PB) CD56(low) NK cells from 60 patients, including 23 subjects with predefined clonal (n = 9) and polyclonal (n = 14) CD56(low) NK-cell expansions, and 37 with CLPD-NK of undetermined clonality; also, PB samples from 10 healthy adults were included. Clonality was established using the human androgen receptor (HUMARA) assay. Clonal NK cells were found to show decreased expression of CD7, CD11b and CD38, and higher CD2, CD94 and HLADR levels vs. normal NK cells, together with a restricted repertoire of expression of the CD158a, CD158b and CD161 killer-associated receptors. In turn, NK cells from both clonal and polyclonal CLPD-NK showed similar/overlapping phenotypic profiles, except for high and more homogeneous expression of CD94 and HLADR, which was restricted to clonal CLPD-NK. We conclude that the CD94(hi)/HLADR+ phenotypic profile proved to be a useful surrogate marker for NK-cell clonality.