Thrombotic genetic risk factors and warfarin pharmacogenetic variants in São Miguel's healthy population (Azores)

<p>Summary</p> <p>Background</p> <p>The Azorean population presents the highest standardized mortality rate for cardiovascular diseases (CVD) when compared to mainland Portugal and other populations. Since thrombosis is a common cause of CVD, we assessed four polymorphisms in three thrombotic risk genes – <it>F5 </it>(G1691A), <it>F2 </it>(G20210A) and <it>MTHFR </it>(C677T, A1298C), in 469 healthy blood donors from São Miguel Island (Azores). We also analysed the <it>CYP2C9 </it>(C430T, A1075C) and <it>VKORC1 </it>(G1639A) variants in fifty-eight individuals with predisposition to thrombosis (possessing at least one variation in <it>F5 </it>or <it>F2 </it>genes and one in <it>MTHFR</it>) to evaluate their warfarin drug response genetic profiles.</p> <p>Results</p> <p>Among the 469 individuals, the data showed that thrombotic risk allele frequencies – 1691A (4.9%), 20210A (1.8%), 677T (41.7%) and 1298C (24.8%) – were similar to other Caucasians, but significantly different from mainland Portuguese (χ², <it>p </it>< 0.001). The combined analysis of these variants identified twenty-two different genetic profiles (genotype order: <it>F5</it>, <it>F2</it>, <it>MTHFR </it>C677T and A1298C). Complete homozygosity for all wild-type alleles (GG GG CC AA) was present in 11.7%, being GG GG CT AA (22.4%) the most frequent profile. The results also demonstrated that 12.4% (58 out of 469) of São Miguel islanders have increased genetic predisposition to thrombosis. Subsequently, we evaluated these individuals for their warfarin response genetic profiles. The data showed that seven out of fifty-eight individuals are poor metabolizers (two with <it>CYP2C9</it>*2/*2 and five with <it>CYP2C9</it>*2/*3 genotypes). <it>VKORC1 </it>polymorphism analysis identified twelve individuals (20.7%) with AA genotype, who probably will require lower doses of warfarin. The joint analysis of <it>CYP2C9 </it>and <it>VKORC1 </it>revealed that 79.3% (46 out of 58) of the individuals carry at least one polymorphism in these genes. Within these, twenty-five individuals (43.1%) need intermediate and/or low doses of warfarin, if treatment is started.</p> <p>Conclusion</p> <p>The present study demonstrated, for the first time, that São Miguel, and possibly the Azores population, shows significant differences on allele frequencies of thrombotic risk factors when compared to mainland Portugal. This research constitutes a primary approach for future studies on CVD, as well as for the implementation of warfarin dosing protocols using the patient's genotypic information.</p

Diagnosis of Human Leptospirosis in a Clinical Setting: Real-Time PCR High Resolution Melting Analysis for Detection of Leptospira at the Onset of Disease:

Currently, direct detection of Leptospira can be done in clinical laboratories by conventional and by real-time PCR (qRT-PCR). We tested a biobank of paired samples of serum and urine from the same patient (202 patients) presenting at the hospital in an area endemic for leptospirosis using qRT-PCR followed by high resolution melting (HRM) analysis. The results were compared with those obtained by conventional nested PCR and with the serologic gold standard microscopic agglutination test (MAT). Differences were resolved by sequencing. qRT-PCR-HRM was positive for 46 of the 202 patients (22.7%, accuracy 100%) which is consistent with known prevalence of leptospirosis in the Azores. MAT results were positive for 3 of the 46 patients (6.5%). Analysis of paired samples allowed us to identify the illness point at which patients presented at the hospital: onset, dissemination or excretion. The melting curve analysis of Leptospira species revealed that 60.9% (28/46) of patients were infected with L. interrogans and 39.1% (18/46) were infected with L. borgpetersenii, both endemic to the Azores. We validated the use of qRT-PCR-HRM for diagnosis of leptospirosis and for identification of the Leptospira species at the earliest onset of infection in a clinical setting, in less than 2 hours.publishersversionpublishe

Evaluation of reference-based two-color methods for measurement of gene expression ratios using spotted cDNA microarrays

BACKGROUND: Spotted cDNA microarrays generally employ co-hybridization of fluorescently-labeled RNA targets to produce gene expression ratios for subsequent analysis. Direct comparison of two RNA samples in the same microarray provides the highest level of accuracy; however, due to the number of combinatorial pair-wise comparisons, the direct method is impractical for studies including large number of individual samples (e.g., tumor classification studies). For such studies, indirect comparisons using a common reference standard have been the preferred method. Here we evaluated the precision and accuracy of reconstructed ratios from three indirect methods relative to ratios obtained from direct hybridizations, herein considered as the gold-standard. RESULTS: We performed hybridizations using a fixed amount of Cy3-labeled reference oligonucleotide (RefOligo) against distinct Cy5-labeled targets from prostate, breast and kidney tumor samples. Reconstructed ratios between all tissue pairs were derived from ratios between each tissue sample and RefOligo. Reconstructed ratios were compared to (i) ratios obtained in parallel from direct pair-wise hybridizations of tissue samples, and to (ii) reconstructed ratios derived from hybridization of each tissue against a reference RNA pool (RefPool). To evaluate the effect of the external references, reconstructed ratios were also calculated directly from intensity values of single-channel (One-Color) measurements derived from tissue sample data collected in the RefOligo experiments. We show that the average coefficient of variation of ratios between intra- and inter-slide replicates derived from RefOligo, RefPool and One-Color were similar and 2 to 4-fold higher than ratios obtained in direct hybridizations. Correlation coefficients calculated for all three tissue comparisons were also similar. In addition, the performance of all indirect methods in terms of their robustness to identify genes deemed as differentially expressed based on direct hybridizations, as well as false-positive and false-negative rates, were found to be comparable. CONCLUSION: RefOligo produces ratios as precise and accurate as ratios reconstructed from a RNA pool, thus representing a reliable alternative in reference-based hybridization experiments. In addition, One-Color measurements alone can reconstruct expression ratios without loss in precision or accuracy. We conclude that both methods are adequate options in large-scale projects where the amount of a common reference RNA pool is usually restrictive

Compreensão acerca dos fatores de risco de lesão por pressão em idosos internados em unidade de terapia intensiva

The aim of this study is to identify and analyze the main risk factors for pressure injuries related to hospitalization in the Intensive Care Unit. This is a bibliographical review of exploratory, quantitative and qualitative scientific literature. To select the sample, a search was made of the electronic databases LILACS and BDENF, from which a careful analysis was carried out, enabling the selection of twenty articles to be used to identify the risks of pressure injury in the elderly in the intensive care unit. The data was collected between March and May of this year. It is understood that there is a high probability of PPI developing in elderly patients after 60 years of age. The average length of hospitalization and pathologies such as hypertension and diabetes mellitus are key factors in the appearance of PI. According to the publications researched, it can be concluded that PPLs are a problem in hospital units that care for critically ill patients due to the permanent or temporary reduction in motility that exists in many cases.El objetivo de este estudio es identificar y analizar los principales factores de riesgo de las lesiones por presión relacionadas con la hospitalización en la Unidad de Cuidados Intensivos. Se trata de una revisión bibliográfica de la literatura científica exploratoria, cuantitativa y cualitativa. Para seleccionar la muestra, se realizó una búsqueda en las bases de datos electrónicas LILACS y BDENF, a partir de la cual se llevó a cabo un cuidadoso análisis que permitió seleccionar veinte artículos para identificar los riesgos de lesiones por presión en ancianos en la Unidad de Cuidados Intensivos. Los datos se recogieron entre marzo y mayo de este año. Se entiende que existe una alta probabilidad de que se desarrolle una IPP en pacientes ancianos después de los 60 años. La duración media de la hospitalización y patologías como la hipertensión y la diabetes mellitus son factores clave en la aparición de la IPP. Según las publicaciones investigadas, se puede concluir que las IPP son un problema común en las unidades hospitalarias que atienden a pacientes críticos debido a la reducción permanente o temporal de la motilidad que existe en muchos casos.Este trabalho tem como objetivo identificar e analisar os principais fatores de risco para lesão por pressão, relacionados a internação em Unidade de Terapia Intensiva. Trata-se de uma revisão bibliográfica em literatura científica exploratória, quantitativa e qualitativa. Para a seleção da amostragem foi feita uma busca a partir dos bancos de dados eletrônicos LILACS e BDENF, a partir disso foi realizada uma análise criteriosa que possibilitou através de uma leitura a seleção de vinte artigos para serem utilizados na identificação dos riscos de lesão por pressão em idosos na unidade de terapia intensiva. Os dados foram coletados entre os meses março a maio deste ano. Entende-se uma alta probabilidade de crescimento de LPP em pacientes idosos após 60 anos de idade. O tempo médio de internação e as patologias como hipertensão arterial e diabetes mellitus apresenta-se como fator primordial ao aparecimento de LPP. De acordo com as publicações pesquisadas, conclui-se que as LPP são agravos presentes em unidades hospitalares que cuidam de pacientes críticos devido a redução permanente ou temporária de motilidade existente em muitos casos. &nbsp

Panorama da Tireoidite de Hashimoto: bases patogênicas, diagnósticas e terapêuticas

Introduction: Hashimoto's thyroiditis (HT) is an autoimmune disease that affects the thyroid gland, designated by the inflammation and destruction of thyroid tissue, triggered by the autoimmune response, which results in the production of autoantibodies, such as antithyroglobulin and antithyroperoxidase antibodies. Objective: To evaluate the pathogenesis, diagnosis and management of Hashimoto's Thyroiditis. Methodology: This is a bibliographic review that included original articles and systematic reviews in English and Portuguese, which addressed the pathogenic, diagnostic and therapeutic aspects of HT, published between 2014 and 2024, selected from the PubMed, Scopus and SciELO databases. After careful selection, 21 articles were chosen to compose this bibliographic review. Results: The pathogenesis of HT presents several relevant aspects, such as the abnormality of regulatory T cells, the exacerbated activity of Th17 cells and the increased expression of inflammatory components and pro-inflammatory cytokines. The diagnosis is made mainly by measuring TSH, free T4 and Anti-TPO, however other complementary tests may be necessary. Management is mainly associated with hormone replacement in patients with hypothyroidism. Considerations: HT is a complex condition, including multiple pathogenic mechanisms that need to be further elucidated. The diagnosis is mainly based on the serum dosage of some compounds. Pharmacological disciplines are associated with the exogenous use of thyroid hormone and surgery in some specific cases.Introdução: A tireoidite de Hashimoto (HT) é uma doença autoimune que afeta a glândula tireoide, caracterizada pela progressiva inflamação e destruição do tecido tireoidiano, desencadeada pela resposta autoimune, que resulta na produção de autoanticorpos, como os anticorpos antitireoglobulina e antitireoperoxidase. Objetivo: Avaliar a patogênese, o diagnóstico e o manejo da Tireoidite de Hashimoto. Metodologia: Trata-se de uma revisão bibliográfica que incluiu artigos originais e revisões sistemáticas em inglês e português, que abordaram os aspectos patogênicos, diagnósticos e terapêuticos da HT, publicados entre 2014 e 2024, selecionados nas bases de dados PubMed, Scopus e SciELO. Após a seleção criteriosa, foram escolhidos 28 artigos para compor esta revisão bibliográfica. Resultados: A patogênese da TH apresenta múltiplos aspectos relevantes, como a anomalia das células T reguladoras, a atividade exacerbada das células Th17 e a expressão aumentada de componentes inflamassômicos e citocinas pró-inflamatórias. O diagnóstico é realizado principalmente pela dosagem de TSH, T4 livre e Anti-TPO, porém pode ser necessário outros exames complementares. O manejo é principalmente associado à reposição hormonal em pacientes com hipotireiodismo. Considerações: A TH é uma condição complexa, incluindo múltiplos mecanismos patogênicos que precisam ser mais elucidados. O diagnóstico é baseado principalmente na dosagem sérica de alguns compostos. As intervenções farmacológicas são associadas ao uso exógeno do hormônio tireoidiano e a cirurgia em alguns casos específicos

SARS-CoV-2 introductions and early dynamics of the epidemic in Portugal

Genomic surveillance of SARS-CoV-2 in Portugal was rapidly implemented by the National Institute of Health in the early stages of the COVID-19 epidemic, in collaboration with more than 50 laboratories distributed nationwide. Methods By applying recent phylodynamic models that allow integration of individual-based travel history, we reconstructed and characterized the spatio-temporal dynamics of SARSCoV-2 introductions and early dissemination in Portugal. Results We detected at least 277 independent SARS-CoV-2 introductions, mostly from European countries (namely the United Kingdom, Spain, France, Italy, and Switzerland), which were consistent with the countries with the highest connectivity with Portugal. Although most introductions were estimated to have occurred during early March 2020, it is likely that SARS-CoV-2 was silently circulating in Portugal throughout February, before the first cases were confirmed. Conclusions Here we conclude that the earlier implementation of measures could have minimized the number of introductions and subsequent virus expansion in Portugal. This study lays the foundation for genomic epidemiology of SARS-CoV-2 in Portugal, and highlights the need for systematic and geographically-representative genomic surveillance.We gratefully acknowledge to Sara Hill and Nuno Faria (University of Oxford) and Joshua Quick and Nick Loman (University of Birmingham) for kindly providing us with the initial sets of Artic Network primers for NGS; Rafael Mamede (MRamirez team, IMM, Lisbon) for developing and sharing a bioinformatics script for sequence curation (https://github.com/rfm-targa/BioinfUtils); Philippe Lemey (KU Leuven) for providing guidance on the implementation of the phylodynamic models; Joshua L. Cherry (National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health) for providing guidance with the subsampling strategies; and all authors, originating and submitting laboratories who have contributed genome data on GISAID (https://www.gisaid.org/) on which part of this research is based. The opinions expressed in this article are those of the authors and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government. This study is co-funded by Fundação para a Ciência e Tecnologia and Agência de Investigação Clínica e Inovação Biomédica (234_596874175) on behalf of the Research 4 COVID-19 call. Some infrastructural resources used in this study come from the GenomePT project (POCI-01-0145-FEDER-022184), supported by COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Lisboa Portugal Regional Operational Programme (Lisboa2020), Algarve Portugal Regional Operational Programme (CRESC Algarve2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF), and by Fundação para a Ciência e a Tecnologia (FCT).info:eu-repo/semantics/publishedVersio

Prevalence, associated factors and outcomes of pressure injuries in adult intensive care unit patients: the DecubICUs study

Funder: European Society of Intensive Care Medicine; doi: http://dx.doi.org/10.13039/501100013347Funder: Flemish Society for Critical Care NursesAbstract: Purpose: Intensive care unit (ICU) patients are particularly susceptible to developing pressure injuries. Epidemiologic data is however unavailable. We aimed to provide an international picture of the extent of pressure injuries and factors associated with ICU-acquired pressure injuries in adult ICU patients. Methods: International 1-day point-prevalence study; follow-up for outcome assessment until hospital discharge (maximum 12 weeks). Factors associated with ICU-acquired pressure injury and hospital mortality were assessed by generalised linear mixed-effects regression analysis. Results: Data from 13,254 patients in 1117 ICUs (90 countries) revealed 6747 pressure injuries; 3997 (59.2%) were ICU-acquired. Overall prevalence was 26.6% (95% confidence interval [CI] 25.9–27.3). ICU-acquired prevalence was 16.2% (95% CI 15.6–16.8). Sacrum (37%) and heels (19.5%) were most affected. Factors independently associated with ICU-acquired pressure injuries were older age, male sex, being underweight, emergency surgery, higher Simplified Acute Physiology Score II, Braden score 3 days, comorbidities (chronic obstructive pulmonary disease, immunodeficiency), organ support (renal replacement, mechanical ventilation on ICU admission), and being in a low or lower-middle income-economy. Gradually increasing associations with mortality were identified for increasing severity of pressure injury: stage I (odds ratio [OR] 1.5; 95% CI 1.2–1.8), stage II (OR 1.6; 95% CI 1.4–1.9), and stage III or worse (OR 2.8; 95% CI 2.3–3.3). Conclusion: Pressure injuries are common in adult ICU patients. ICU-acquired pressure injuries are associated with mainly intrinsic factors and mortality. Optimal care standards, increased awareness, appropriate resource allocation, and further research into optimal prevention are pivotal to tackle this important patient safety threat