Uma carreira cientifica só se faz de experiência. E quem não a tem, de alguém a tem que aprender.... more Uma carreira cientifica só se faz de experiência. E quem não a tem, de alguém a tem que aprender. Por isso, não posso deixar de transmitir um profundo obrigado aos que com o seu apoio feito de exemplo, exigência, profissionalismo e amizade, tornaram possível levar a bom termo o trabalho aqui apresentado: à Prof. Doutora Maria de Sousa pela orientação directa dos trabalhos desta dissertação e à Prof. Doutora Maria João Saraiva pela generosidade com que aceitou ser co-orientadora desta tese. Gostaria ainda de agradecer a todos os que no Mestrado de Imunologia, com generosidade me foram treinando a capacidade de pensar, planear, testar e analisar. Uma palavra especial para a Doutora Graça Porto pelo acompanhamento na fase de planeamento de experiências e análise dos resultados. Em segundo lugar não posso deixar de mencionar o Doutor António Silva, pelo apoio que permitiu fundar os alicerces da futura construção. À D. Rosa Lacerda, o meu obrigado pelo sempre presente e eficiente apoio no laboratório. Finalmente à Doutora Berta Martins e à D. Helena Branco o meu agradecimento pelo trabalho de tipagem HLA. À Prof. Doutora Maria de Sousa quero ainda expressar o meu agradecimento pelas oportunidades que me proporcionou de trabalhar com alguns dos melhores grupos científicos na área do repertório da célula T. Aos directores desses grupos Doutores David Posnett, Hans Wigzell e Rolf Hultcrantz, o meu agradecimento, por me terem aceite nos seus laboratórios, ensinando-me técnicas e métodos de trabalho sem os quais esta tese seria certamente diferente. Uma palavra especial ainda para a Doutora Dude Gigliotti e o Dr. Rolland Andersson, que de muito perto me apoiaram no meu trabalho em Estocolmo. Gostaria ainda de agradecer a todos os que colaboraram na elaboração do texto da presente dissertação: à Prof. Doutora Maria de Sousa e à Doutora Graça Porto, pelas repetidas e muito produtivas sessões de discussão do trabalho, resultados e suas implicações, bem como pelas inúmeras vezes que se dignaram realizar uma leitura critica do manuscrito. Na leitura critica do manuscrito colaboraram ainda a Dr. a Luciana Pinho e o Dr. Fernando Arosa. Finalmente, gostaria de agradecer à Dr. a Margarida Ribeiro, pela tradução do resumo da dissertação para a língua francesa. Mas a ciência é em ultima análise uma forma de cultura. Uma cultura com valores próprios que importa alimentar desde muito cedo. A cultura cientifica não é no entanto apanágio apenas de cientistas, mas de todos os que têm como valor supremo a verdade, e como ambição a sua descoberta. Assim, não posso deixar de considerar que o caminho que me conduziu até esta dissertação só foi possível graças ao extraordinário trabalho educativo que os meus pais desenvolveram. Primeiro, ensinando-me a considerar a verdade como um bem essencial, depois apoiando-me até ás ultimas consequências no caminhar por terrenos que não podiam compreender, apenas apoiar, e finalmente por terem sabido partilhar a minha excitação por cada passo dado nesse percurso de interrogações feito. Neste aspecto, os meus agradecimentos vão também para os meus irmãos, e sobretudo a Esmeralda. Sem o seu apoio, tantas vezes incondicional, esta tese não estaria ainda certamente terminada. O presente trabalho teve o apoio financeiro da JNICT, do Dr. Peter Oppenheimer, Fundo APBR, e programas Ciência e Praxis XXI. Ao Instituto de Ciências Biomédicas Abel Salazar, quero agradecer por me ter facultado os meios físicos para a realização de parte dos trabalhos laboratoriais.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common erythrocyte enzymopathy, a... more Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common erythrocyte enzymopathy, affecting over 400 million people worldwide. Portugal is a low prevalence country, but immigrants from endemic regions are common, particularly in the south of the country. In the present study, we report the laboratory findings observed in two black proband children with low G6PD enzyme activity (23 and 18%). The study also included their first-degree relatives. Both biochemical parameters (enzyme activity, electrophoretic mobility and cytochemical test) and genetic determinations (mutation and haplotype characterisation) were performed.
Rhinovirus is a common picornavirus with over 150 serotypes and three species, which is responsib... more Rhinovirus is a common picornavirus with over 150 serotypes and three species, which is responsible for half of the human common cold cases. In people with chronic respiratory conditions and elders, it may also cause life-threatening diseases. Transmission routes are not definitively established but may involve direct human-to-human and indirect transmission (surfaces and aerosols based). In the present study, year-long presence of virus was tested by qPCR in the nostrils of young healthy volunteers and indoor and outdoor air samples. Results were correlated to atmospheric conditions (meteorological and air quality parameters) and voluntaries immune system-related genetic polymorphisms (TOLLIP rs5743899, IL6 rs1800795, IL1B rs16944, TNFA rs1800629) typed by PCR-RFLP. Nasal samples showed increased frequency and viral titers of Rhinovirus in spring and autumn. No indoor air samples tested positive for Rhinovirus, whereas outdoor air samples tested positive in late autumn. Sun radiation, atmospheric SO 2 , and benzene levels correlated with nostrils Rhinovirus detection. Both IL6 and TOLLIP polymorphisms but not TNFA or IL1B influenced Rhinovirus detection in the nostrils of voluntaries. Taken together, the results indicate that Rhinovirus circulation is determined by environmental conditions (weather, airborne virus, and air pollution) and genetically encoded individual variation in immunity.
Methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from a general hospital in ... more Methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from a general hospital in Oporto, Portugal, during two periods (1992-1993 and 1996-2000) were characterized by pulsed-field gel electrophoresis (PFGE) of SmaI fragments, and by hybridization of ClaI digests with mecA and Tn554 probes, discriminating the isolates in mecA::Tn554::PFGE genotypes. In addition, a representative sample of the defined genotypes was characterized by multilocus sequence typing (MLST) and SCCmec (staphylococcal cassette chromosome mec) typing, generating the corresponding ST-SCCmec types. In 1992-1993, 77% of MRSA belonged to the Iberian clone (genotype I::E::A or ST247-IA). In 1996-2000, the frequency of this clone decreased to 19% and the majority (69%) of the isolates belonged to another international clone, the Brazilian MRSA (genotype XI::B::B or ST239-IIIA). Trimethoprim/sulfamethoxazole (SXT) was confirmed to be an important phenotypic marker to distinguish the Iberian (SXT-susceptible) and the Brazilian (SXT-resistant) clones in MRSA isolates from Portugal. Our observations document major shifts in the dominant MRSA clonal types that occurred in this hospital since 1992, suggesting a selective advantage of the Brazilian relatively to the Iberian clone. In addition to these two MRSA clones that are the most frequent in Portuguese hospitals since the early 1990s, sporadic MRSA clones (representing 14% of the total) were identified and characterized.
Mutation analysis was performed for two HFE mutations (C282Y, H63D) in unrelated patients with he... more Mutation analysis was performed for two HFE mutations (C282Y, H63D) in unrelated patients with hereditary haemochromatosis (n ¼ 92), family members of patients (n ¼ 34), and unrelated controls (n ¼ 157) from Northern Germany. 87/92 patients (94•6%) revealed the C282Y mutation in homozygous form, five were heterozygous. No H63D mutation was found in 174 chromosomes of patients homozygous for C282Y, whereas four of the heterozygote patients also carried the H63D mutation. Among the control group, 9•6% were heterozygotes for C282Y. 2/157 subjects were homozygous, 37/157 were heterozygous for the H63D mutation, but showed no signs of iron overload.
Genetics of MHC 231 Methods: The study group consisted of 45 individuals with a history of heavy ... more Genetics of MHC 231 Methods: The study group consisted of 45 individuals with a history of heavy alcohol consumption. In average, the daily alcohol intake was 200 g (range = 100-400) for an average pedod of 35 years. All individuals were phenotyped by flow cytometry for CD4, CD8 and CD28 expression in freshly collected peripheral blood. A control group of 30 healthy individuals without history of alcohol intake was also studied. Results: Analysis of the CD28 expression revealed that the frequently observed increase in CD8+ T cell proportions in this population of heavy drinkers (22/45 = 49%) were mainly due to expansions of CD28-cells. This observation contrasted with the results in the control population where the CD8+ T call proportions were dependant on beth CD28+ and CD28-calls. Finally we analysed the influence of WA-H on the expansions of CD8+CD28-T cells and observed a significant association between CD8+CD28-cell numbers and the presence of the H63D mutation (p = 0.05). The mean ± SD values for the CD8+CD28populations were: 20.1 ± 6.0 in homozygous for the mutation, 16.5 ± 3.6. in heterozygous and 11.9 ± 1.4 in individuals with the normal gane. Conclusion: The present results led us to conclude that the presence of the HLA-H H63D mutation in heavy drinkers may somehow contribute to the expansion of CD8+CD28-T calls which, in tum, may have a protective role in the toxicity of alcohol.
] LISIO COSTA ci al desigual, com países com uma freqüência que atinge os 25% e outros cuja freq... more ] LISIO COSTA ci al desigual, com países com uma freqüência que atinge os 25% e outros cuja freqüência é inferior a 0,5%2. Portugal é um país de baixa prevalência, cerca de 0,51 %6, sendo no entanto de registar uma crescente imigração de áreas endémicas, nomeadamente de ...
We report a case of a patient with two B-cell lymphoproliferative disorders: CD5 ؊ /CD23 ؉ B-cell... more We report a case of a patient with two B-cell lymphoproliferative disorders: CD5 ؊ /CD23 ؉ B-cell chronic lymphocytic leukemia and CD5 ؉ /CD23 ؊ mantle cell lymphoma. These disorders were diagnosed simultaneously based on flow cytometry, immunohistochemistry, fluorescence in situ hybridization, and polymerase chain reaction-based molecular studies. The B-cell lymphocytic leukemia clone predominated in the blood and bone marrow, whereas the mantle cell clone predominated in lymph nodes. Cytometry Part B (Clin. Cytometry) 51B:41-44, 2003.
We characterized the genetic nature of bthalassaemia in northern Portugal. Of the 164 patients st... more We characterized the genetic nature of bthalassaemia in northern Portugal. Of the 164 patients studied three were b-thalassaemia major cases (one IVS-1-6/ bЊ39 and two homozygous IVS-1-110). The analysis of the frequency of each mutation in the families revealed that the codon 6(¹A) mutation was unexpectedly frequent (40%) and associated with the b-globin haplotype E, and not with the usual European and North African CD6(¹A) haplotypes. In contrast, the frequency of IVS-1-6 (8%) and bЊ39 (19%) was found to be lower than in the rest of the country. The frequency of all other mutations was similar to previous reports for central/southern Portugal. Six families carried none of the most frequent mutations in the Mediterranean area. These families were studied by gene sequencing, revealing that three families carried a previously described mutation (CD16 G → A). The remaining families carried previously unidentified mutations: one showed an 86 bp insertion in exon 2 (named HGSA) and two showed a deletion of a cytidine in codon 11 (CD11(¹C)). The results, showing a high frequency (82%) of bЊ mutations, strongly indicates that genetic counselling should be intensified as a means of preventing the spread of the severe mutations found.
Purpose: We examined the significance of the CAG repeat polymorphism in the pathogenesis of crypt... more Purpose: We examined the significance of the CAG repeat polymorphism in the pathogenesis of cryptorchidism. Materials and Methods: Genomic deoxyribonucleic acid (DNA) was extracted from blood samples from 42 cryptorchid boys and from 31 non-cryptorchid control subjects. In the cryptorchid group, 7 had bilateral cryptorchidism and 6 had patent processus vaginalis in the contralateral side. To determine the number of CAG repeats, the DNA was amplified by polymerase chain reaction and sequenced. Results: The mean CAG repeat length in the AR gene was 22.5 (range 16 to 28) in patients and 21.5 (range 17 to 26) in controls (non-significant). Patients with bilateral cryptorchidism had a mean length of 24.3 (range 21 to 26) and patients with unilateral cryptorchidism and patent processus vaginalis in the contra lateral side had a mean of 25.2 (range 21 to 28), which was statistically different from controls (p = 0.015 and p = 0.005 respectively). Conclusion: CAG repeat length of the AR gene does not seem to play a major role in patients with unilateral cryptorchidism. However, in patients with bilateral undescended testis, a less functional androgen receptor through a longer polyglutamine chain may have a role in its pathogenesis. In the same way, patients with unilateral cryptorchidism a contralateral patent processus vaginalis have longer CAG repeats that might be responsible for a slower testicular descent and incomplete closure of the processus vaginalis.
Pathophysiology of Haemostasis and Thrombosis, 2002
Since FV-Leiden polymorphism was first described in 1994, a growing number of polymorphic loci ha... more Since FV-Leiden polymorphism was first described in 1994, a growing number of polymorphic loci have been identified in association with increased genetic risk for thrombophilia. Often however, these risk factors have been studied in isolation of the remaining known phenotype linked polymorphisms. This fact has, at least in part, been justified by the laborious techniques traditionally used in the genotyping studies, as well as its relatively high costs. Another major problem concerning these studies has been the non-negligible incidence of dubious genotypes, resulting from the manual, labour intensive techniques applied, and their sometimes difficult to read output's. These difficulties have also hampered the widespread use of genotyping data in the clinical assessment of the genetic risk levels both in patients and their relatives, leaving some clinicians less than convinced about its clinical usefulness. Recently however, the introduction of new genetic techniques in the clinical genetics laboratory has started to change this picture. Most notably, the advent of Realtime-PCR has brought the possibility of genotyping patients and controls at a large scale, with increased specificity, automation and speed. Moreover, the use of these techniques in the clinical genetics setting has not only increased the quality of the results, but most
Aim To quantify bacterial equivalents before and after chemomechanical preparation using 3% sodiu... more Aim To quantify bacterial equivalents before and after chemomechanical preparation using 3% sodium hypochlorite (NaOCl) and intracanal dressing with calcium hydroxide paste (Ca(OH) 2) or 2% Chlorhexidine digluconate gel (CHX) in necrotic pulps associated or not with apical periodontitis and to further compare this quantification with counts of anaerobic microorganisms. Methodology Prospective clinical trial in 69 singlerooted adult teeth (strict inclusion criteria); CHX group: 34; Ca(OH) 2 group: 35. Bacteria samples were taken at baseline (S1), after chemomechanical preparation (S2) and after 14 days of intracanal dressing (S3). Bacterial equivalents were assessed by broad-range real-time polymerase chain reaction (qPCR), and live viable bacteria measured with conventional anaerobic culture (CFU/mL). Descriptive/inferential analysis was performed with SPSS vs. 20.0 (a = 0.05) using the Kruskal-Wallis, Mann-Whitney and chi-squared tests and Spearman's correlation coefficients. Results Both groups showed a significant decrease between S1 and S2 (Mann-Whitney U-test; P < 0.001) both in qPCR and in culture. In the Ca (OH) 2-group, no variation was observed between S2 and S3 by qPCR and culture. In contrast, the CHX group showed a significant increase from S2 to S3 by both techniques. The two groups were only significantly different in S3 (Mann-Whitney U-test; P 0.001), with a worse performance in the CHX group. Again, these results were congruent by both approaches. Data from both approaches correlate reasonably (r S < 0.5). Conclusions Infected root canals contained a high bacterial load, and the chemomechanical root canal preparation reduced bacterial equivalents by 99.1% and anaerobic counts by 98.5%. Intracanal dressings were not efficient at reducing bacterial load, but the 14-day intracanal dressing with Ca(OH) 2 performed significantly better than CHX, particularly in cases with apical periodontitis.
Resumo tem-se verificado um aumento de alergias em todo o mundo estimando-se que 10-20% dos portu... more Resumo tem-se verificado um aumento de alergias em todo o mundo estimando-se que 10-20% dos portugueses sofra de rinite alérgica (rA). no presente estudo procuramos caracterizar a sintomatologia e percepção de rA em estudantes e trabalhadores da indústria Química. Dos resultados destacamos que o número de inquiridos que afirmam sofrer de rA (19,9%) é semelhante ao descrito na literatura, mas um número maior de inquiridos apresenta sintomatologia sugestiva de rA. Assim, a rA encontra-se provavelmente sub-diagnosticada, sendo possivelmente confundida com gripe ou constipação.
Revista Portuguesa de Estomatologia, Medicina Dentária e Cirurgia Maxilofacial, Apr 1, 2014
Objectives: Assessment of bacterial reduction after chemo-mechanical preparation (using 3% sodium... more Objectives: Assessment of bacterial reduction after chemo-mechanical preparation (using 3% sodium hypochlorite) with or without intracanal dressing (calcium hydroxide paste (Ca(OH) 2) or 2% chlorhexidine digluconate gel (CHX)) in necrotic pulps associated or not with apical lesion. Methods: Prospective clinical trial, in 69 adult patient's teeth with pulpal necrosis associated or not with apical periodontitis. Microbiological root-canal-sampling occurred before treatment (S1), after chemo-mechanical preparation (S2) and after 14 days intracanal dressing (S3). Colony Forming Units (CFU) were counted after growth in aerobic, anaerobic and microaerofilic cultures. Comparison of the median CFUs treatments and culture media was done with the Friedman test. Comparison of the intracanal dressing effect at S3 was done with the Wilcoxon and the Mann-Whitney tests. Because of the huge differences in bacterial counts variations were expressed as log 10 to analyze differences among intracanal medication groups. S2 and S3 counts were expressed as percentage of CFU reduction regarding S1 counts. Results: Significant differences were detected between S1, S2 and S3 (Friedman test; p < 0.001), showing a significant decrease from S1 to S2 (Wilcoxon test; p < 0.004), followed by a significant increase from S2 to S3 (p < 0.001) for the CHX group, maintenance for the Ca(OH) 2 group in aerobic/anaerobic (Wilcoxon test; p = 0.777/0.227), and increase in the microaerofilic culture (Wilcoxon test; p = 0.047). The two groups only differed significantly in S3 (Mann-Whitney test; p ≤ 0.001), with a worse performance in the CHX group. Conclusions: Treatment significantly reduced the number of bacteria but failed to render all root canals sterile. Ca(OH) 2 performed better than CHX gel.
I female) and 10 heterozygotic individuais. They were refered to two Northem Central Hospitais (H... more I female) and 10 heterozygotic individuais. They were refered to two Northem Central Hospitais (Hospital Geral Santo Antonio and Hospital de Crianças Maria Pia). Of the affected individuais, three were classified as class I and ten as class IIL according to the WHO.
Enfermedades Infecciosas Y Microbiologia Clinica, 2004
We evaluated three rapid methods to detect methicillin-resistant Staphylococcus aureus (MRSA) and... more We evaluated three rapid methods to detect methicillin-resistant Staphylococcus aureus (MRSA) and compared them with PCR amplification of mecA. A total of 103 S. aureus strains were studied by MRSA-Screen, BBL Crystal, Velogene Genomic and mecA PCR. All the methods detected the 61 MRSA strains having the mecA gene, showing 100% sensitivity and specificity. Despite the correlation between all the rapid methods and PCR, the ease of use and shorter turnaround time of MRSA-Screen were important factors leading to the selection of this method as the routine screening technique for MRSA.
Pathobionts, particularly Mycobacterium avium subsp. paratuberculosis (MAP) and Escherichia coli ... more Pathobionts, particularly Mycobacterium avium subsp. paratuberculosis (MAP) and Escherichia coli isolates with adherence/invasive ability (AIEC) have been associated with inflammatory bowel disease (IBD), particularly Crohn’s disease (CD). This study aimed to evaluate the frequency of viable MAP and AIEC in a cohort of IBD patients. As such, MAP and E. coli cultures were established from faecal and blood samples (with a total n = 62 for each) of patients with CD (n = 18), ulcerative colitis (UC, n = 15), or liver cirrhosis (n = 7), as well as from healthy controls (HC, n = 22). Presumptive positive cultures were tested by polymerase chain reaction (PCR), for a positive confirmation of MAP or E. coli identity. E. coli-confirmed isolates were then tested for AIEC identity using adherence and invasion assays in the epithelial cell line of Caco-2 and survival and replication assays in the macrophage cell line of J774. MAP sub-culture and genome sequencing were also performed. MAP was mo...
Uma carreira cientifica só se faz de experiência. E quem não a tem, de alguém a tem que aprender.... more Uma carreira cientifica só se faz de experiência. E quem não a tem, de alguém a tem que aprender. Por isso, não posso deixar de transmitir um profundo obrigado aos que com o seu apoio feito de exemplo, exigência, profissionalismo e amizade, tornaram possível levar a bom termo o trabalho aqui apresentado: à Prof. Doutora Maria de Sousa pela orientação directa dos trabalhos desta dissertação e à Prof. Doutora Maria João Saraiva pela generosidade com que aceitou ser co-orientadora desta tese. Gostaria ainda de agradecer a todos os que no Mestrado de Imunologia, com generosidade me foram treinando a capacidade de pensar, planear, testar e analisar. Uma palavra especial para a Doutora Graça Porto pelo acompanhamento na fase de planeamento de experiências e análise dos resultados. Em segundo lugar não posso deixar de mencionar o Doutor António Silva, pelo apoio que permitiu fundar os alicerces da futura construção. À D. Rosa Lacerda, o meu obrigado pelo sempre presente e eficiente apoio no laboratório. Finalmente à Doutora Berta Martins e à D. Helena Branco o meu agradecimento pelo trabalho de tipagem HLA. À Prof. Doutora Maria de Sousa quero ainda expressar o meu agradecimento pelas oportunidades que me proporcionou de trabalhar com alguns dos melhores grupos científicos na área do repertório da célula T. Aos directores desses grupos Doutores David Posnett, Hans Wigzell e Rolf Hultcrantz, o meu agradecimento, por me terem aceite nos seus laboratórios, ensinando-me técnicas e métodos de trabalho sem os quais esta tese seria certamente diferente. Uma palavra especial ainda para a Doutora Dude Gigliotti e o Dr. Rolland Andersson, que de muito perto me apoiaram no meu trabalho em Estocolmo. Gostaria ainda de agradecer a todos os que colaboraram na elaboração do texto da presente dissertação: à Prof. Doutora Maria de Sousa e à Doutora Graça Porto, pelas repetidas e muito produtivas sessões de discussão do trabalho, resultados e suas implicações, bem como pelas inúmeras vezes que se dignaram realizar uma leitura critica do manuscrito. Na leitura critica do manuscrito colaboraram ainda a Dr. a Luciana Pinho e o Dr. Fernando Arosa. Finalmente, gostaria de agradecer à Dr. a Margarida Ribeiro, pela tradução do resumo da dissertação para a língua francesa. Mas a ciência é em ultima análise uma forma de cultura. Uma cultura com valores próprios que importa alimentar desde muito cedo. A cultura cientifica não é no entanto apanágio apenas de cientistas, mas de todos os que têm como valor supremo a verdade, e como ambição a sua descoberta. Assim, não posso deixar de considerar que o caminho que me conduziu até esta dissertação só foi possível graças ao extraordinário trabalho educativo que os meus pais desenvolveram. Primeiro, ensinando-me a considerar a verdade como um bem essencial, depois apoiando-me até ás ultimas consequências no caminhar por terrenos que não podiam compreender, apenas apoiar, e finalmente por terem sabido partilhar a minha excitação por cada passo dado nesse percurso de interrogações feito. Neste aspecto, os meus agradecimentos vão também para os meus irmãos, e sobretudo a Esmeralda. Sem o seu apoio, tantas vezes incondicional, esta tese não estaria ainda certamente terminada. O presente trabalho teve o apoio financeiro da JNICT, do Dr. Peter Oppenheimer, Fundo APBR, e programas Ciência e Praxis XXI. Ao Instituto de Ciências Biomédicas Abel Salazar, quero agradecer por me ter facultado os meios físicos para a realização de parte dos trabalhos laboratoriais.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common erythrocyte enzymopathy, a... more Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common erythrocyte enzymopathy, affecting over 400 million people worldwide. Portugal is a low prevalence country, but immigrants from endemic regions are common, particularly in the south of the country. In the present study, we report the laboratory findings observed in two black proband children with low G6PD enzyme activity (23 and 18%). The study also included their first-degree relatives. Both biochemical parameters (enzyme activity, electrophoretic mobility and cytochemical test) and genetic determinations (mutation and haplotype characterisation) were performed.
Rhinovirus is a common picornavirus with over 150 serotypes and three species, which is responsib... more Rhinovirus is a common picornavirus with over 150 serotypes and three species, which is responsible for half of the human common cold cases. In people with chronic respiratory conditions and elders, it may also cause life-threatening diseases. Transmission routes are not definitively established but may involve direct human-to-human and indirect transmission (surfaces and aerosols based). In the present study, year-long presence of virus was tested by qPCR in the nostrils of young healthy volunteers and indoor and outdoor air samples. Results were correlated to atmospheric conditions (meteorological and air quality parameters) and voluntaries immune system-related genetic polymorphisms (TOLLIP rs5743899, IL6 rs1800795, IL1B rs16944, TNFA rs1800629) typed by PCR-RFLP. Nasal samples showed increased frequency and viral titers of Rhinovirus in spring and autumn. No indoor air samples tested positive for Rhinovirus, whereas outdoor air samples tested positive in late autumn. Sun radiation, atmospheric SO 2 , and benzene levels correlated with nostrils Rhinovirus detection. Both IL6 and TOLLIP polymorphisms but not TNFA or IL1B influenced Rhinovirus detection in the nostrils of voluntaries. Taken together, the results indicate that Rhinovirus circulation is determined by environmental conditions (weather, airborne virus, and air pollution) and genetically encoded individual variation in immunity.
Methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from a general hospital in ... more Methicillin-resistant Staphylococcus aureus (MRSA) isolates recovered from a general hospital in Oporto, Portugal, during two periods (1992-1993 and 1996-2000) were characterized by pulsed-field gel electrophoresis (PFGE) of SmaI fragments, and by hybridization of ClaI digests with mecA and Tn554 probes, discriminating the isolates in mecA::Tn554::PFGE genotypes. In addition, a representative sample of the defined genotypes was characterized by multilocus sequence typing (MLST) and SCCmec (staphylococcal cassette chromosome mec) typing, generating the corresponding ST-SCCmec types. In 1992-1993, 77% of MRSA belonged to the Iberian clone (genotype I::E::A or ST247-IA). In 1996-2000, the frequency of this clone decreased to 19% and the majority (69%) of the isolates belonged to another international clone, the Brazilian MRSA (genotype XI::B::B or ST239-IIIA). Trimethoprim/sulfamethoxazole (SXT) was confirmed to be an important phenotypic marker to distinguish the Iberian (SXT-susceptible) and the Brazilian (SXT-resistant) clones in MRSA isolates from Portugal. Our observations document major shifts in the dominant MRSA clonal types that occurred in this hospital since 1992, suggesting a selective advantage of the Brazilian relatively to the Iberian clone. In addition to these two MRSA clones that are the most frequent in Portuguese hospitals since the early 1990s, sporadic MRSA clones (representing 14% of the total) were identified and characterized.
Mutation analysis was performed for two HFE mutations (C282Y, H63D) in unrelated patients with he... more Mutation analysis was performed for two HFE mutations (C282Y, H63D) in unrelated patients with hereditary haemochromatosis (n ¼ 92), family members of patients (n ¼ 34), and unrelated controls (n ¼ 157) from Northern Germany. 87/92 patients (94•6%) revealed the C282Y mutation in homozygous form, five were heterozygous. No H63D mutation was found in 174 chromosomes of patients homozygous for C282Y, whereas four of the heterozygote patients also carried the H63D mutation. Among the control group, 9•6% were heterozygotes for C282Y. 2/157 subjects were homozygous, 37/157 were heterozygous for the H63D mutation, but showed no signs of iron overload.
Genetics of MHC 231 Methods: The study group consisted of 45 individuals with a history of heavy ... more Genetics of MHC 231 Methods: The study group consisted of 45 individuals with a history of heavy alcohol consumption. In average, the daily alcohol intake was 200 g (range = 100-400) for an average pedod of 35 years. All individuals were phenotyped by flow cytometry for CD4, CD8 and CD28 expression in freshly collected peripheral blood. A control group of 30 healthy individuals without history of alcohol intake was also studied. Results: Analysis of the CD28 expression revealed that the frequently observed increase in CD8+ T cell proportions in this population of heavy drinkers (22/45 = 49%) were mainly due to expansions of CD28-cells. This observation contrasted with the results in the control population where the CD8+ T call proportions were dependant on beth CD28+ and CD28-calls. Finally we analysed the influence of WA-H on the expansions of CD8+CD28-T cells and observed a significant association between CD8+CD28-cell numbers and the presence of the H63D mutation (p = 0.05). The mean ± SD values for the CD8+CD28populations were: 20.1 ± 6.0 in homozygous for the mutation, 16.5 ± 3.6. in heterozygous and 11.9 ± 1.4 in individuals with the normal gane. Conclusion: The present results led us to conclude that the presence of the HLA-H H63D mutation in heavy drinkers may somehow contribute to the expansion of CD8+CD28-T calls which, in tum, may have a protective role in the toxicity of alcohol.
] LISIO COSTA ci al desigual, com países com uma freqüência que atinge os 25% e outros cuja freq... more ] LISIO COSTA ci al desigual, com países com uma freqüência que atinge os 25% e outros cuja freqüência é inferior a 0,5%2. Portugal é um país de baixa prevalência, cerca de 0,51 %6, sendo no entanto de registar uma crescente imigração de áreas endémicas, nomeadamente de ...
We report a case of a patient with two B-cell lymphoproliferative disorders: CD5 ؊ /CD23 ؉ B-cell... more We report a case of a patient with two B-cell lymphoproliferative disorders: CD5 ؊ /CD23 ؉ B-cell chronic lymphocytic leukemia and CD5 ؉ /CD23 ؊ mantle cell lymphoma. These disorders were diagnosed simultaneously based on flow cytometry, immunohistochemistry, fluorescence in situ hybridization, and polymerase chain reaction-based molecular studies. The B-cell lymphocytic leukemia clone predominated in the blood and bone marrow, whereas the mantle cell clone predominated in lymph nodes. Cytometry Part B (Clin. Cytometry) 51B:41-44, 2003.
We characterized the genetic nature of bthalassaemia in northern Portugal. Of the 164 patients st... more We characterized the genetic nature of bthalassaemia in northern Portugal. Of the 164 patients studied three were b-thalassaemia major cases (one IVS-1-6/ bЊ39 and two homozygous IVS-1-110). The analysis of the frequency of each mutation in the families revealed that the codon 6(¹A) mutation was unexpectedly frequent (40%) and associated with the b-globin haplotype E, and not with the usual European and North African CD6(¹A) haplotypes. In contrast, the frequency of IVS-1-6 (8%) and bЊ39 (19%) was found to be lower than in the rest of the country. The frequency of all other mutations was similar to previous reports for central/southern Portugal. Six families carried none of the most frequent mutations in the Mediterranean area. These families were studied by gene sequencing, revealing that three families carried a previously described mutation (CD16 G → A). The remaining families carried previously unidentified mutations: one showed an 86 bp insertion in exon 2 (named HGSA) and two showed a deletion of a cytidine in codon 11 (CD11(¹C)). The results, showing a high frequency (82%) of bЊ mutations, strongly indicates that genetic counselling should be intensified as a means of preventing the spread of the severe mutations found.
Purpose: We examined the significance of the CAG repeat polymorphism in the pathogenesis of crypt... more Purpose: We examined the significance of the CAG repeat polymorphism in the pathogenesis of cryptorchidism. Materials and Methods: Genomic deoxyribonucleic acid (DNA) was extracted from blood samples from 42 cryptorchid boys and from 31 non-cryptorchid control subjects. In the cryptorchid group, 7 had bilateral cryptorchidism and 6 had patent processus vaginalis in the contralateral side. To determine the number of CAG repeats, the DNA was amplified by polymerase chain reaction and sequenced. Results: The mean CAG repeat length in the AR gene was 22.5 (range 16 to 28) in patients and 21.5 (range 17 to 26) in controls (non-significant). Patients with bilateral cryptorchidism had a mean length of 24.3 (range 21 to 26) and patients with unilateral cryptorchidism and patent processus vaginalis in the contra lateral side had a mean of 25.2 (range 21 to 28), which was statistically different from controls (p = 0.015 and p = 0.005 respectively). Conclusion: CAG repeat length of the AR gene does not seem to play a major role in patients with unilateral cryptorchidism. However, in patients with bilateral undescended testis, a less functional androgen receptor through a longer polyglutamine chain may have a role in its pathogenesis. In the same way, patients with unilateral cryptorchidism a contralateral patent processus vaginalis have longer CAG repeats that might be responsible for a slower testicular descent and incomplete closure of the processus vaginalis.
Pathophysiology of Haemostasis and Thrombosis, 2002
Since FV-Leiden polymorphism was first described in 1994, a growing number of polymorphic loci ha... more Since FV-Leiden polymorphism was first described in 1994, a growing number of polymorphic loci have been identified in association with increased genetic risk for thrombophilia. Often however, these risk factors have been studied in isolation of the remaining known phenotype linked polymorphisms. This fact has, at least in part, been justified by the laborious techniques traditionally used in the genotyping studies, as well as its relatively high costs. Another major problem concerning these studies has been the non-negligible incidence of dubious genotypes, resulting from the manual, labour intensive techniques applied, and their sometimes difficult to read output's. These difficulties have also hampered the widespread use of genotyping data in the clinical assessment of the genetic risk levels both in patients and their relatives, leaving some clinicians less than convinced about its clinical usefulness. Recently however, the introduction of new genetic techniques in the clinical genetics laboratory has started to change this picture. Most notably, the advent of Realtime-PCR has brought the possibility of genotyping patients and controls at a large scale, with increased specificity, automation and speed. Moreover, the use of these techniques in the clinical genetics setting has not only increased the quality of the results, but most
Aim To quantify bacterial equivalents before and after chemomechanical preparation using 3% sodiu... more Aim To quantify bacterial equivalents before and after chemomechanical preparation using 3% sodium hypochlorite (NaOCl) and intracanal dressing with calcium hydroxide paste (Ca(OH) 2) or 2% Chlorhexidine digluconate gel (CHX) in necrotic pulps associated or not with apical periodontitis and to further compare this quantification with counts of anaerobic microorganisms. Methodology Prospective clinical trial in 69 singlerooted adult teeth (strict inclusion criteria); CHX group: 34; Ca(OH) 2 group: 35. Bacteria samples were taken at baseline (S1), after chemomechanical preparation (S2) and after 14 days of intracanal dressing (S3). Bacterial equivalents were assessed by broad-range real-time polymerase chain reaction (qPCR), and live viable bacteria measured with conventional anaerobic culture (CFU/mL). Descriptive/inferential analysis was performed with SPSS vs. 20.0 (a = 0.05) using the Kruskal-Wallis, Mann-Whitney and chi-squared tests and Spearman's correlation coefficients. Results Both groups showed a significant decrease between S1 and S2 (Mann-Whitney U-test; P < 0.001) both in qPCR and in culture. In the Ca (OH) 2-group, no variation was observed between S2 and S3 by qPCR and culture. In contrast, the CHX group showed a significant increase from S2 to S3 by both techniques. The two groups were only significantly different in S3 (Mann-Whitney U-test; P 0.001), with a worse performance in the CHX group. Again, these results were congruent by both approaches. Data from both approaches correlate reasonably (r S < 0.5). Conclusions Infected root canals contained a high bacterial load, and the chemomechanical root canal preparation reduced bacterial equivalents by 99.1% and anaerobic counts by 98.5%. Intracanal dressings were not efficient at reducing bacterial load, but the 14-day intracanal dressing with Ca(OH) 2 performed significantly better than CHX, particularly in cases with apical periodontitis.
Resumo tem-se verificado um aumento de alergias em todo o mundo estimando-se que 10-20% dos portu... more Resumo tem-se verificado um aumento de alergias em todo o mundo estimando-se que 10-20% dos portugueses sofra de rinite alérgica (rA). no presente estudo procuramos caracterizar a sintomatologia e percepção de rA em estudantes e trabalhadores da indústria Química. Dos resultados destacamos que o número de inquiridos que afirmam sofrer de rA (19,9%) é semelhante ao descrito na literatura, mas um número maior de inquiridos apresenta sintomatologia sugestiva de rA. Assim, a rA encontra-se provavelmente sub-diagnosticada, sendo possivelmente confundida com gripe ou constipação.
Revista Portuguesa de Estomatologia, Medicina Dentária e Cirurgia Maxilofacial, Apr 1, 2014
Objectives: Assessment of bacterial reduction after chemo-mechanical preparation (using 3% sodium... more Objectives: Assessment of bacterial reduction after chemo-mechanical preparation (using 3% sodium hypochlorite) with or without intracanal dressing (calcium hydroxide paste (Ca(OH) 2) or 2% chlorhexidine digluconate gel (CHX)) in necrotic pulps associated or not with apical lesion. Methods: Prospective clinical trial, in 69 adult patient's teeth with pulpal necrosis associated or not with apical periodontitis. Microbiological root-canal-sampling occurred before treatment (S1), after chemo-mechanical preparation (S2) and after 14 days intracanal dressing (S3). Colony Forming Units (CFU) were counted after growth in aerobic, anaerobic and microaerofilic cultures. Comparison of the median CFUs treatments and culture media was done with the Friedman test. Comparison of the intracanal dressing effect at S3 was done with the Wilcoxon and the Mann-Whitney tests. Because of the huge differences in bacterial counts variations were expressed as log 10 to analyze differences among intracanal medication groups. S2 and S3 counts were expressed as percentage of CFU reduction regarding S1 counts. Results: Significant differences were detected between S1, S2 and S3 (Friedman test; p < 0.001), showing a significant decrease from S1 to S2 (Wilcoxon test; p < 0.004), followed by a significant increase from S2 to S3 (p < 0.001) for the CHX group, maintenance for the Ca(OH) 2 group in aerobic/anaerobic (Wilcoxon test; p = 0.777/0.227), and increase in the microaerofilic culture (Wilcoxon test; p = 0.047). The two groups only differed significantly in S3 (Mann-Whitney test; p ≤ 0.001), with a worse performance in the CHX group. Conclusions: Treatment significantly reduced the number of bacteria but failed to render all root canals sterile. Ca(OH) 2 performed better than CHX gel.
I female) and 10 heterozygotic individuais. They were refered to two Northem Central Hospitais (H... more I female) and 10 heterozygotic individuais. They were refered to two Northem Central Hospitais (Hospital Geral Santo Antonio and Hospital de Crianças Maria Pia). Of the affected individuais, three were classified as class I and ten as class IIL according to the WHO.
Enfermedades Infecciosas Y Microbiologia Clinica, 2004
We evaluated three rapid methods to detect methicillin-resistant Staphylococcus aureus (MRSA) and... more We evaluated three rapid methods to detect methicillin-resistant Staphylococcus aureus (MRSA) and compared them with PCR amplification of mecA. A total of 103 S. aureus strains were studied by MRSA-Screen, BBL Crystal, Velogene Genomic and mecA PCR. All the methods detected the 61 MRSA strains having the mecA gene, showing 100% sensitivity and specificity. Despite the correlation between all the rapid methods and PCR, the ease of use and shorter turnaround time of MRSA-Screen were important factors leading to the selection of this method as the routine screening technique for MRSA.
Pathobionts, particularly Mycobacterium avium subsp. paratuberculosis (MAP) and Escherichia coli ... more Pathobionts, particularly Mycobacterium avium subsp. paratuberculosis (MAP) and Escherichia coli isolates with adherence/invasive ability (AIEC) have been associated with inflammatory bowel disease (IBD), particularly Crohn’s disease (CD). This study aimed to evaluate the frequency of viable MAP and AIEC in a cohort of IBD patients. As such, MAP and E. coli cultures were established from faecal and blood samples (with a total n = 62 for each) of patients with CD (n = 18), ulcerative colitis (UC, n = 15), or liver cirrhosis (n = 7), as well as from healthy controls (HC, n = 22). Presumptive positive cultures were tested by polymerase chain reaction (PCR), for a positive confirmation of MAP or E. coli identity. E. coli-confirmed isolates were then tested for AIEC identity using adherence and invasion assays in the epithelial cell line of Caco-2 and survival and replication assays in the macrophage cell line of J774. MAP sub-culture and genome sequencing were also performed. MAP was mo...
Uploads
Papers by José Cabeda