A survey of entomopathogenic nematode species in continental Portugal

Entomopathogenic nematodes (EPN) are lethal parasites of insects, used as biocontrol agents. The objectives of this work were to survey the presence of EPN in continental Portugal and to characterize the different species. Of the 791 soil samples collected throughout continental Portugal, 53 were positive for EPN. Steinernema feltiae and Heterorhabditis bacteriophora were the two most abundant species. Analysis of EPN geographical distribution revealed an association between nematode species and vegetation type. Heterorhabditis bacteriophora was mostly found in the Alentejo region while S. feltiae was present in land occupied by agriculture with natural vegetation, broadleaved forest, mixed forest and transitional woodland-shrub, agro-forestry areas, complex cultivated patterns and non-irrigated arable land. Although no clear association was found between species and soil type, S. feltiae was typically recovered from cambisols and H. bacteriophora was more abundant in lithosols. Sequencing of the internal transcribed spacer (ITS) region indicated that S. feltiae was the most abundant species, followed by H. bacteriophora. Steinernema intermedium and S. kraussei were each isolated from one site and Steinernema sp. from two sites. Phylogenetic analyses of ITS, D2D3 expansion region of the 28S rRNA gene, as well as mitochondrial cytochrome c oxidase subunit I (COXI) and cytochrome b (cytb) genes, was performed to evaluate the genetic diversity of S. feltiae and H. bacteriophora. No significant genetic diversity was found among H. bacteriophora isolates. However, COXI seems to be the best marker to study genetic diversity of S. feltiae. This survey contributes to the understanding of EPN distribution in Europe

Novel bio-strategies for the detoxification of mycotoxins using lactic acid bacteria

A ocorrência de bolores micotoxigénicos pertencentes aos géneros Aspergillus, Penicillium e Fusarium em alimentos para consumo Humano e animal, tem um impacto importante sobre a saúde pública e constitui também um importante problema económico. Isto é devido à síntese por este tipo de fungos filamentosos de metabolitos altamente tóxicos conhecidos como micotoxinas. A maioria das micotoxinas são substâncias cancerígenas, mutagénicas, neurotóxicas e imunossupressoras, sendo a ocratoxina A (OTA) uma das mais importantes. A OTA é uma micotoxina, tóxica para os animais e Humanos principalmente devido às suas propriedades nefrotóxicas. Alguns grupos de bactérias gram positivas nomeadamente as bactérias do ácido láctico (BAL) são capazes de controlar o crescimento de fungos, melhorando e aumentando a vida útil de muitos produtos fermentados e, assim, reduzir os riscos para a saúde provocados pela exposição às micotoxinas. Algumas BAL são, também, capazes de destoxificar certas micotoxinas. Em trabalhos anteriores do nosso grupo foi observada a biodegradação da OTA por estirpes de Pediococcus parvulus isoladas de vinhos do Douro. Assim, com este trabalho, pretendeu-se compreender com maior detalhe o processo de biodegradação da OTA pelas referidas estirpes e identificar quais as enzimas que estão associadas à sua biodegradação. Para atingir este objetivo utilizaram-se algumas ferramentas ioinformáticas (BLAST, CLUSTALX2, CLC Sequence Viewer 7, Finch TV), desenharam-se primers específicos e realizaram-se PCR específicos para os genes envolvidos. Através da utilização de ferramentas de bioinformática, foi possível identificar várias proteínas que pertencem à família das carboxipeptidases e que podem eventualmente participar no processo da degradação da OTA, tais como D-Ala-D-Ala carboxipeptidase serínica e carboxipeptidase membranar. Estas BAL podem desempenhar um papel importante na destoxificação da OTA, sendo as carboxipeptidases uma das enzimas envolvidas na sua biodegradação.The occurrence of mycotoxigenic moulds such as Aspergillus, Penicillium and Fusarium in food and feed has an important impact on public health and also constitutes a major economic problem. This is due through the synthesis of highly toxic metabolites known as mycotoxins by these kind of moulds. Most of the mycotoxins are carcinogenic, mutagenic, neurotoxic and immunosuppressive, being ochratoxin A (OTA) one of the most important. OTA is toxic to animals and Humans, mainly due to its nephrotoxic properties. Some group of Gram-positive bacteria named lactic acid bacteria (LAB) are able to control moulds ́ growth, improving the shelf life of many fermented products and reducing health risks due to exposure to mycotoxins. Some LAB are capable of mycotoxin detoxification. From previous work of our group was observed OTA biodegradation by Pediococcus parvulus isolated from Douro wines. So, we wanted to have a better understanding of this OTA degradation process and identify which molecules where present in this process. For achieving our aim we used some bioinformatics tools (BLAST, CLUSTALX2, CLC Sequence Viewer 7, Finch TV), we designed specific primers and we did gene specific PCR. Through the employment of bioinformatics tools it was possible to identify several proteins belonging to the carboxypeptidase family that participate in the process of OTA degradation, such as serine type D-Ala-D-Ala carboxypeptidase and membrane carboxypeptidase. LAB can play an important role in the mycotoxins detoxification, being carboxypeptidase proteins one of the molecules present in the OTA degradation process.Agradecimentos: Este trabalho foi financiado por fundos FEDER através do Programa Operacional Factores de Competitividade - COMPETE e por fundos nacionais através da Fundação para a Ciência e a Tecnologia -FCT, ref. FCOMP-01-0124-FEDER-028029 e PTDC/AGR-TEC/3900/2012, respetivamente. Luís Abrunhosa recebeu apoio através da bolsa Incentivo/EQB/LA0023/2014 do ON.2 O Novo Norte

Translation and Validation of the Patient Activation Measure in Portuguese People with Type 2 Diabetes Mellitus

Introduction: Management of diabetes mellitus is largely dependent on patients' active participation in care. The 'Patient Activation Measure 13' assesses patients' knowledge, skills, and confidence in self-care. We aimed to translate, culturally adapt, and validate the 'Patient Activation Measure 13' to Portuguese, in people with type 2 diabetes. Material and Methods: The translation and cultural adaptation occurred in six phases. A convenience sample of people with type 2 diabetes was recruited from the waiting rooms of a diabetes outpatient centre in Lisbon, between March and April 2014. The questionnaire was self-administered; medical records were reviewed to obtain glycated haemoglobin levels. Main statistical analyses were based on the Rasch rating scale model. Results: The response rate for the final questionnaire was 76%. Rasch analysis was conducted on 193 respondents. Respondents had a mean age of 67.1 (SD 10.1) years, 42.7% were women, and the mean patient activation measure score (0 - 100) in the sample was 58.5 (SD 10.1). The sample was low to moderate in terms of activation: 40.4% were low in activation (levels 1 and 2), 49.7% were in level 3, and 9.8% were in level 4, the highest level of activation. All items had good fit and the response categories functioned well. Item reliability was 0.97 and person reliability was between 0.77 (real) and 0.83 (model). Discussion: The 'Patient Activation Measure 13' was translated and culturally adapted to European Portuguese and validated in people with diabetes, showing good psychometric properties. Future research should aim at evaluating test-retest reliability of the Portuguese 'Patient Activation Measure 13', and exploring its ability to measure changes in activation over time. Conclusion: The 'Patient Activation Measure 13' is now available in European Portuguese and has good psychometric properties.Liliana Laranjo was supported by a Junior Clinical Research award from the Harvard Medical School-Portugal program (HMSP-ICJ/0005/2010; Fundacao para a Ciencia e Tecnologia)

a comprehensive survey of medical doctor's perspectives in Portugal

Publisher Copyright: Copyright: © 2023 Pedro et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Artificial Intelligence (AI) is increasingly influential across various sectors, including healthcare, with the potential to revolutionize clinical practice. However, risks associated with AI adoption in medicine have also been identified. Despite the general understanding that AI will impact healthcare, studies that assess the perceptions of medical doctors about AI use in medicine are still scarce. We set out to survey the medical doctors licensed to practice medicine in Portugal about the impact, advantages, and disadvantages of AI adoption in clinical practice. We designed an observational, descriptive, cross-sectional study with a quantitative approach and developed an online survey which addressed the following aspects: impact on healthcare quality of the extraction and processing of health data via AI; delegation of clinical procedures on AI tools; perception of the impact of AI in clinical practice; perceived advantages of using AI in clinical practice; perceived disadvantages of using AI in clinical practice and predisposition to adopt AI in professional activity. Our sample was also subject to demographic, professional and digital use and proficiency characterization. We obtained 1013 valid, fully answered questionnaires (sample representativeness of 99%, confidence level (p< 0.01), for the total universe of medical doctors licensed to practice in Portugal). Our results reveal that, in general terms, the medical community surveyed is optimistic about AI use in medicine and are predisposed to adopt it while still aware of some disadvantages and challenges to AI use in healthcare. Most medical doctors surveyed are also convinced that AI should be part of medical formation. These findings contribute to facilitating the professional integration of AI in medical practice in Portugal, aiding the seamless integration of AI into clinical workflows by leveraging its perceived strengths according to healthcare professionals. This study identifies challenges such as gaps in medical curricula, which hinder the adoption of AI applications due to inadequate digital health training. Due to high professional integration in the healthcare sector, particularly within the European Union, our results are also relevant for other jurisdictions and across diverse healthcare systems.publishersversionpublishe

Positive Fluid Balance Was Associated with Mortality in Patients with Acute-on-Chronic Liver Failure: a Cohort Study

Purpose: We aimed to study the effect of FB in the outcomes of critically-ill patients with cirrhosis. Materials: Retrospective analysis of all adult consecutive admissions of patients with cirrhosis and organ failures to the Intensive Care Unit (ICU) at Curry Cabral Hospital (Lisbon, Portugal) and University of Alberta Hospital (Edmonton, Canada) on 08/2013-08/2017. Primary exposure was FB at 3 and 7 days post ICU admission. Primary endpoint was hospital mortality. Results: Amongst 333 patients, median age was 56 years and 67.6% were men. Median MELD, APACHEII, CLIF-SOFA, and CLIF-C-ACLF scores on ICU admission were 27, 28, 14, and 54, respectively. ICU and hospital mortality rates were 33.0% and 49.2%, respectively. While median FB at 3 days post ICU admission (+5.46 l vs. +6.62 l; P = 0.74) was not associated with hospital mortality, higher median FB at 7 days post ICU admission (+13.50 l vs. +6.90 l; P = 0.036) was associated with higher hospital mortality. This association remained significant (OR 95%CI = 1.04 [1.01;1.07] per each l) after adjustment for confounders (age, ascites, infection, lactate, and number of organ failures). Conclusions: FB may be a therapeutic target that helps to improve the outcomes of patients with acute-on-chronic liver failure. This data may inform future clinical trials.info:eu-repo/semantics/publishedVersio

Bio-detoxification of mycotoxins by lactic acid bacteria from different food matrices

Lactic acid bacteria (LAB) play a key role in the biopreservation of a wide range of fermented food products, such as yogurt, cheese, fermented milks, meat, fish, vegetables (sauerkraut, olives and pickles), certain beer brands, wines and silage, allowing their safe consumption, which gave to these bacteria a GRAS (Generally Recognised as Safe) status. Besides that, the use of LAB in food and feed is a promising strategy to reduce the exposure to dietary mycotoxins, improving their shelf life and reducing health risks, given the unique mycotoxin decontaminating characteristic of some LAB. Mycotoxins present carcinogenic, mutagenic, teratogenic, neurotoxic and immunosuppressive effects over animals and Humans, being the most important ochratoxin A (OTA), aflatoxins (AFB1), trichothecenes, zearalenone (ZEA), fumonisin (FUM) and patulin. In a previous work of our group it was observed OTA biodegradation by some strains of Pediococcus parvulus isolated from Douro wines. So, the aim of this study was to enlarge the screening of the biodetoxification over more mycotoxins besides OTA, including AFB1, and ZEA. This ability was checked in a collection of LAB isolated from vegetable (wine, olives, fruits and silage) and animal (milk and dairy products, sausages) sources. All LAB strains were characterized phenotypically (Gram, catalase) and genotypically. Molecular characterisation of all LAB strains was performed using genomic fingerprinting by MSP- PCR with (GTG)5 and csM13 primers. The identification of the isolates was confirmed by 16S rDNA sequencing. To study the ability of LAB strains to degrade OTA, AFB1 and ZEA, a MRS broth medium was supplemented with 2.0 g/mL of each mycotoxin. For each strain, 2 mL of MRS supplemented with the mycotoxins was inoculated in triplicate with 109 CFU/mL. The culture media and bacterial cells were extracted by the addition of an equal volume of acetonitrile/methanol/acetic acid (78:20:2 v/v/v) to the culture tubes. A 2 mL sample was then collected and filtered into a clean 2 mL vial using PP filters with 0.45 m pores. The samples were preserved at 4 °C until HPLC analysis. Among LAB tested, 10 strains isolated from milk were able to eliminate AFB1, belonging to Lactobacillus casei (7), Lb. paracasei (1), Lb. plantarum (1) and 1 to Leuconostoc mesenteroides. Two strains of Enterococcus faecium and one of Ec. faecalis from sausage eliminated ZEA. Concerning to strains of vegetal origin, one Lb. plantarum isolated from elderberry fruit, one Lb. buchnerii and one Lb. parafarraginis both isolated from silage eliminated ZEA. Other 2 strains of Lb. plantarum from silage were able to degrade both ZEA and OTA, and 1 Lb. buchnerii showed activity over AFB1. These enzymatic activities were also verified genotypically through specific gene PCR and posteriorly confirmed by sequencing analysis. In conclusion, due the ability of some strains of LAB isolated from different sources to eliminate OTA, AFB1 and ZEA one can recognize their potential biotechnological application to reduce the health hazards associated with these mycotoxins. They may be suitable as silage inoculants or as feed additives or even in food industry.Acknowledgements This work was funded by FEDER funds through the COMPETE and by national funds through FCT, Ref. FCOMP-01-0124-FEDER-028029 and PTDC/AGR-T EC/3900/2012, respectively. Luís Abrunhosa received support through grant Incentivo/EQB/LA0023/2014 from ON.2 O Novo Nort

Effects of soil management practices and irrigation on plant water relations and productivity of chestnut stands under Mediterranean conditions

The effects of different soil management practices and irrigation on plant water relations, physiological response and productivity of chestnut stands in Northeastern Portugal were assessed during four growing seasons (2003 to 2006). Treatments were: conventional soil tillage up to 15–20 cm depth with a tine cultivator thrice a year (CT); no tillage with spontaneous herbaceous vegetation (NV); no tillage with rainfed seeded pasture (NP); and no tillage with irrigated seeded pasture (NIP). Results suggest that soil water availability was the most critical parameter for chestnut productivity over the study period. In all treatments, high predawn leaf water potentials (-0.40 to -0.55 MPa) were observed during the dry seasons of 2003, 2004 and 2006, showing no critical conditions for plant productivity, which is ascribed to water availability in deep soil layers. In contrast, in 2005, an extremely dry year, water potentials decreased and varied from -1.46 to - 1.72 MPa in late summer, showing unfavourable conditions for nut production. Maintenance of spontaneous herbaceous vegetation without irrigation enhanced productivity of chestnut stands as compared with the conventional tillage system and the no tillage system with seeded pasture. Productivity in the soil watering system (NIP treatment) was not significantly different from that observed in the NV treatment. Therefore, studies on the irrigation strategy should be developed, in order to increase its efficiency especially in stands with young tree

Tilt Training Increases Vasoconstrictor Reserve in Patients with Neurocardiogenic Syncope

A síncope neurocardiogénica (SNc) é uma entidade clínica comum, resultante de uma resposta autonómica reflexa excessiva durante o stress ortostático. As diferentes opções terapêuticas são controversas e de eficácia limitada. O treino de ortostatismo (TTr) tem-se mostrado uma alternativa prometedora no tratamento destes doentes (D). No entanto, permanece por esclarecer o seu mecanismo de acção e o impacto clínico numa população com SNc recorrente. Objectivo: Caracterizar a resposta hemodinâmica e autonómica durante um programa de TTr em doentes com SNc refratária às medidas convencionais. População e métodos: Foram estudados 28D (50% do sexo masculino, 41±14 anos), sem evidência de cardiopatia, com SNc documentada em teste de ortostatismo passivo. O TTr incluiu 9 sessões hospitalares (3x/semana, 30 minutos) com monitorizac¸ão contínua de pressão arterial e frequência cardíaca (60◦ - 6 sessões - 70◦ - 3 sessões), complementadas com treino diário no domicílio e elevação da cabeceira a 10◦ durante o sono. O volume sistólico, o débito cardíaco, a resistência vascular periférica, a sensibilidade do barorreflexo e a variabilidade da frequência cardíaca foram calculados. Todos os doentes foram reavaliados no fim do 1.◦ mês e no final de cada 6 meses num período máximo de 36 meses (follow-up 24±12 meses). Resultados: Ao longo das sessões de TTr verificou-se um aumento significativo e consistente da resistência total periférica (1485±225 vs. 1591±187 dyne*s/cm−5, p < 0,05) associado a uma diminuição do seu desvio-padrão (206±60 vs. 150±42, p < 0,05). Durante o período de follow-up, houve recorrência de síncope em 5D (19%), com redução significativa do número de síncopes (4,0±3,2/D nos 12 meses pre-TTr vs. 1,4±0,8/D pos-TTr, p < 0,05). Conclusão: Em doentes com SNc refratária, o TTr mostrou ser uma opção terapêutica eficaz, com benefício a longo prazo. A melhor tolerância ao ortostatismo parece resultar do aumento da reserva vasoconstritora e da sua menor variabilidade

O Treino de Ortostatismo (Tilt Training) Aumenta a Reserva Vasoconstritora em Doentes com Síncope Reflexa Neurocardiogénica

Neurocardiogenic syncope (NCS) is a common clinical entity resulting from an excessive reflex autonomic response, particularly during orthostatism. Treatment options are controversial and of limited effectiveness. Tilt training (TT) is a promising option to treat these patients. However, its mechanism of action and clinical impact remain unclear. OBJECTIVE: To characterize hemodynamic and autonomic responses during a TT program in patients with NCS refractory to conventional measures. METHODS: We studied 28 patients (50% male, mean age 41±14 years) without structural heart disease, with NCS documented by tilt testing. The TT program included 9 tilt sessions (3 times a week, 30 min) (60° - 6 sessions, 70° - 3 sessions), under ECG and blood pressure monitoring combined with home orthostatic self-training and 10° head-up during sleep. Systolic volume, cardiac output, total peripheral resistance, baroreflex sensitivity and heart-rate variability were computed. Patients were reassessed at 1 month and every 6 months for a maximum of 36 months (24±12 months). RESULTS: Over the course of the TT program there was a significant increase in total peripheral resistance (1485±225 vs. 1591±187 dyn·s·cm(-5), p<0.05), with a decrease in standard deviation (206±60 vs. 150±42, p<0.05). During follow-up, syncope recurred in five patients (19%), with a significant reduction in the number of episodes (4.0±3.2/patient in the 12 months before TT vs. 1.4±0.8/patient post-TT, p<0.05). CONCLUSION: In refractory NCS, TT may be an effective therapeutic option, with long-term benefits. These results appear to be due to an increase in vasoconstrictor reserve combined with a reduction in its variance

Effects of Vagal Stimulation on Induction and Termination of Atrial Fibrillation in an in Vivo Rabbit Heart Model

INTRODUCTION: Vagal activity is thought to influence atrial electrophysiological properties and play a role in the initiation and maintenance of atrial fibrillation (AF). In this study, we assessed the effects of acute vagal stimulation (vagus_stim) on atrial conduction times, atrial and pulmonary vein (PV) refractoriness, and vulnerability to induction of AF in the rabbit heart with intact autonomic innervation. METHODS: An open-chest epicardial approach was performed in 11 rabbits (New Zealand; 3.9-5.0 kg), anesthetized and artificially ventilated after neuromuscular blockade. A 3-lead ECG was obtained. Atrial electrograms were recorded along the atria, from right to left (four monopolar electrodes), together with a circular electrode adapted for proximal left PV assessment. Acute vagus nerve stimulation was obtained with bipolar electrodes (20 Hz). Epicardial activation was recorded in sinus rhythm, and the conduction time from right (RA) to left atrium (LA), and from RA to PVs, was measured in basal conditions and during vagus_stim. The atrial effective refractory period (ERP) and dispersion of refractoriness (Disp_A) were analyzed. Vulnerability to AF induction was assessed at the right (RAA) and left (LAA) atrial appendages and the PVs. Atrial stimulation (50 Hz) was performed alone or combined with vagus_stim. Heart rate and blood pressure were monitored. RESULTS: In basal conditions, there was a significant delay in conduction from RA to PVs, not influenced by vagus_stim, and the PV ERPs were shorter than those measured in LA and LAA, but without significant differences compared to RA and RAA. During vagus_stim, conduction times between RA and LA increased from 16+8 ms to 27+6 ms (p 10 s in 45.4% of rabbits during vagus_stim, and ceased after vagus_stim in 4 out of these 5 cases. In 3 animals, PV tachycardia, with fibrillatory conduction, induced with 50 Hz PV pacing during vagus_stim. CONCLUSIONS: Vagus_stim reduces interatrial conduction velocity and significantly shortens atrial ERP, contributing to the induction and duration of AF episodes in the in vivo rabbit heart. This model may be useful for the assessment of autonomic influence on the pathophysiology of AF