Antimalarial resistance risk in Mozambique detected by a novel quadruplex droplet digital PCR assay.

While the Plasmodium falciparum malaria parasite continues to cause severe disease globally, Mozambique is disproportionally represented in malaria case totals. Acquisition of copy number variations (CNVs) in the parasite genome contributes to antimalarial drug resistance through overexpression of drug targets. Of interest, piperaquine resistance is associated with plasmepsin 2 and 3 CNVs (pfpmp2 and pfpmp3, respectively), while CNVs in the multidrug efflux pump, multidrug resistance-1 (pfmdr1), increase resistance to amodiaquine and lumefantrine. These antimalarials are partner drugs in artemisinin combination therapies (ACTs) and therefore, CNV detection with accurate and efficient tools is necessary to track ACT resistance risk. Here, we evaluated ~300 clinically derived samples collected from three sites in Mozambique for resistance-associated CNVs. We developed a novel, medium-throughput, quadruplex droplet digital PCR (ddPCR) assay to simultaneously quantify the copy number of pfpmp3, pfpmp2, and pfmdr1 loci in these clinical samples. By using DNA from laboratory parasite lines, we show that this nanodroplet-based method is capable of detecting picogram levels of parasite DNA, which facilitates its application for low yield and human host-contaminated clinical surveillance samples. Following ddPCR and the application of quality control standards, we detected CNVs in 13 of 229 high-quality samples (prevalence of 5.7%). Overall, our study revealed a low number of resistance CNVs present in the parasite population across all three collection sites, including various combinations of pfmdr1, pfpmp2, and pfpmp3 CNVs. The potential for future ACT resistance across Mozambique emphasizes the need for continued molecular surveillance across the region.

Anti-malarial resistance in Mozambique: Absence of Plasmodium falciparum Kelch 13 (K13) propeller domain polymorphisms associated with resistance to artemisinins.

BACKGROUND: Malaria remains one of the most serious public health problems in sub-Saharan Africa and Mozambique is the world's fourth largest contributor, with 4.7% of disease cases and 3.6% of total deaths due to malaria. Its control relies on the fight against the vector and treatment of confirmed cases with anti-malarial drugs. Molecular surveillance is an important tool for monitoring the spread of anti-malarial drug resistance. METHODS: A cross-sectional study recruited 450 participants with malaria infection detected by Rapid Diagnostic Tests, from three different study sites (Niassa, Manica and Maputo) between April and August 2021. Correspondent blood samples were collected on filter paper (Whatman® FTA® cards), parasite DNA extracted and pfk13 gene sequenced using Sanger method. SIFT software (Sorting Intolerant From Tolerant) was used, predict whether an amino acid substitution affects protein function. RESULTS: No pfkelch13-mediated artemisinin resistance gene mutation was detected in this study settings. However, non-synonymous mutations were detected at prevalence of 10.2%, 6% and 5% in Niassa, Manica and Maputo, respectively. Most (56.3%) of the reported non-synonymous mutations were due to substitution at the first base of the codon, 25% at the second base and 18.8% at the third base. Additionally, 50% of non-synonymous mutations showed a SIFTscore bellow cut off value of 0.05, therefore, they were predicted to be deleterious. CONCLUSION: These results do not show an emergence of artemisinin resistance cases in Mozambique. However, the increased number of novel non-synonymous mutations highlights the relevance of increasing the number of studies focused on the molecular surveillance of artemisinin resistance markers, for its early detection.

Efficacy of proactive long-term maintenance therapy of canine atopic dermatitis with 0.0584% hydrocortisone aceponate spray: a double-blind placebo controlled pilot study.

BACKGROUND: Long-term remission between flares of canine atopic dermatitis (CAD) can be difficult to achieve. Therefore, additional strategic forms of treatment are needed in order to target flare prevention. The concept of proactive therapy is recommended in the European guidelines for the treatment of human atopic eczema. OBJECTIVES: To evaluate the efficacy of a proactive treatment regimen with a 0.0584% hydrocortisone aceponate (HCA) spray for CAD. ANIMALS: Client-owned dogs with spontaneous atopic dermatitis (AD) (n = 41). METHODS: This pilot study was conducted as a randomised, placebo-controlled, double-blinded clinical trial with an end-point of treatment failure. Dogs were treated once daily to remission, then randomly assigned to receive either the HCA spray (n = 21) or a placebo (n = 20) spray on two consecutive days each week. All dogs were on appropriate flea control. No topical or systemic anti-inflammatory or antimicrobial agents were permitted. Intention-to-treat analysis was used. RESULTS: At Day 0, all the dogs were in remission or had mild AD based on their Canine Atopic Dermatitis Extent and Severity Index, version 3 (CADESI-03) scores. The time to relapse was significantly higher in the HCA group (median 115 d; range 31-260 d) compared to the placebo group (median 33 d; range 15-61 d) (P < 0.0001). No adverse events were attributable to the HCA spray. Four dogs were lost to follow-up and four were withdrawn after receiving prohibited medication. CONCLUSIONS AND CLINICAL IMPORTANCE: These results indicate that proactive long-term therapy of CAD with an HCA spray administered on two consecutive days each week is effective and well-tolerated.

Nursing in hemotherapy services: considerations on public policies associated with blood and blood components.

OBJECTIVE: Reflect on how the evolution of legislation related to blood and blood-based products has shaped the role of nurses in hemotherapy services. METHODS: A reflective study, carried out employing surveys in databases, about the evolution of the nurse's role in hemotherapy services. RESULTS: Several legislations, published since 1950, have encouraged blood donation and shaped the work of nurses in hemotherapy services, being the most relevant Law #10.205/2001 (Lei nº 10.205/2001), about blood collection, processing, storage, distribution, and blood transfusion; and COFEN's resolution # 629/2020 (COFEN nº 629/2020), which addresses in detail the practice of these services. FINAL CONSIDERATIONS: The evolution of legislation related to blood and blood-based products contributed to the consolidation of nurses' attributions in hemotherapy services since it guaranteed legal support and defined the practices in these services.

Portuguese Authorship in Published Clinical Trials: Differences in Industry and Investigator Initiated Trials.

INTRODUCTION: The aim of this study was to investigate the Portuguese authorship in publications resulting from trials initiated by the industry or investigators and run in Portugal. MATERIAL AND METHODS: Clinical trials with Portuguese institutions as sponsor or recruiting centers, and registered in four clinical trial registries, in the last 14 years, were assessed. Publications of completed trials, from both the initiative of the industry and investigatorswere screened and compared. RESULTS: The percentage of published trials initiated by industry and investigators was similar (28.0%). However, the percentage of completed investigator-initiated trials (43.6%) was lower when compared to industry trials (69.7%). There was a higher percentage of Portuguese authorship in published investigator-initiated trials when compared with industry-initiated trials (47.1% vs 8.5%, respectively). Moreover, industry-initiated trials with Portuguese authors were published in journals with lower journal impact factor when compared with those published without authorship of Portuguese investigators. Oncology was the therapeutic area with the highest number of clinical trial registrations and publications. However, in publications with Portuguese authors, industry Initiated trials mainly focused on neurology while investigator-initiated trials had a higher number of papers in the fields of gastroenterology and infection diseases. Published trials with Portuguese authorship, initiated by the industry or investigators, also targeted different populations and had different purposes. In both cases, no significant differences were observed in terms of the journal impact factor or in the alignment of the published randomized trials with the respective reporting guidelines. DISCUSSION: When compared with previous publications, this study showed an increasing trend in the number of clinical trials in Portugal, published within similar timeframes, after trial conclusion. Even though both industry and investigator trials are published within the standards for reporting trials, the low number of Portuguese authorships in industry publications might underline the need for invigorating these independent clinical trials in Portugal by capacitating and empowering national clinical research teams. CONCLUSION: This study confirmed that even though all registered trials had the involvement of Portuguese institutions as a recruiting center, not all the published trials had Portuguese investigators as authors, mainly those initiated by the industry.

Introdução: Este estudo teve por objetivo investigar a autoria Portuguesa em publicações que resultem de ensaios clínicos iniciados pela indústria e por investigadores, que tenham decorrido em Portugal. Material e Métodos: Quatro plataformas de registo de ensaios clínicos foram utilizadas para encontrar ensaios clínicos tendo instituições Portuguesas como promotor ou centro de recrutamento nos últimos 14 anos. Foram analisadas e comparadas as publicações dos estudos completos, da iniciativa da indústria e de investigadores Resultados: A percentagem de ensaios da iniciativa da indústria e de investigadores que são publicados era semelhante (~ 28,0%). Porém, a percentagem de ensaios completos da iniciativa de investigadores era mais baixa (43,6%) quando comparada com os ensaios completos da indústria (69,7%). Existiu uma maior percentagem de autores portugueses em ensaios publicados da iniciativa do investigador quando comparado com os ensaios da iniciativa da indústria (47,1% vs 8,5%). Para além disso, ensaios da iniciativa da indústria com autores portugueses foram publicados em jornais com fatores de impacto inferiores quando comparados com aqueles publicados sem autores portugueses. A oncologia foi a área terapêutica com maior número de ensaios registados e publicados. No entanto, em publicações com autores Portugueses, a indústria focou-se sobretudo na neurologia e os investigadores em gastroenterologia e doenças infeciosas. Ensaios publicados com autores portugueses, iniciados tanto pela indústria como por investigadores, focaram-se em populações diferentes e têm propósitos diferentes. Em ambos os casos, não foram encontradas diferenças estatisticamente significativas no fator de impacto dos jornais, nem no alinhamento dos ensaios aleatorizados publicados com as normas sobre escrita de artigos científicos. Discussão: Quando comparado com publicações anteriores, este estudo mostrou uma tendência de crescimento no número de ensaios clínicos em Portugal, sendo publicados em intervalos de tempo semelhantes após a sua conclusão. Embora os ensaios publicados da iniciativa da indústria e de investigadores estejam alinhados com as normas sobre escrita de artigos científicos, o baixo número de autorias nacionais em publicações de ensaios da indústria, sublinha a necessidade de revigorar os ensaios clínicos da iniciativa de investigadores através da capacitação e emancipação das equipas de investigação nacionais. Conclusão: Apesar de todos os ensaios registados terem o envolvimento de instituições portuguesas como centros de recrutamento, nem todos os ensaios têm autores portugueses nas publicações, principalmente aqueles que são iniciados pela indústria.

Nursing in hemotherapy services: considerations on public policies associated with blood and blood components / Enfermería en servicios de hemoterapia: reflexiones acerca de las políticas públicas vueltas a la sangre y hemocomponentes / Enfermagem em serviços de hemoterapia: reflexões acerca das políticas públicas voltadas ao sangue e hemocomponentes

ABSTRACT Objective: Reflect on how the evolution of legislation related to blood and blood-based products has shaped the role of nurses in hemotherapy services. Methods: A reflective study, carried out employing surveys in databases, about the evolution of the nurse's role in hemotherapy services. Results: Several legislations, published since 1950, have encouraged blood donation and shaped the work of nurses in hemotherapy services, being the most relevant Law #10.205/2001 (Lei nº 10.205/2001), about blood collection, processing, storage, distribution, and blood transfusion; and COFEN's resolution # 629/2020 (COFEN nº 629/2020), which addresses in detail the practice of these services. Final considerations: The evolution of legislation related to blood and blood-based products contributed to the consolidation of nurses' attributions in hemotherapy services since it guaranteed legal support and defined the practices in these services.

RESUMEN Objetivo: Reflejar sobre como la evolución de las legislaciones dirigidas a la sangre y hemoderivados moldeó la actuación del enfermero en los servicios de hemoterapia. Métodos: Estudio reflexivo, realizado mediante levantamientos en bases de datos, sobre la evolución de la actuación del enfermero en servicios de hemoterapia. Resultados: Verificado que las diversas legislaciones, publicadas desde 1950, fomentaron la donación de sangre y moldaron la actuación del enfermero en servicios de hemoterapia, siendo las de mayor relevancia: la Ley nº 10.205/2001, sobre la recolecta, procesamiento, almacenaje, distribución y aplicación de la sangre; y la resolución del COFEN n.º 629/2020, que aborda en detalhes la actuación en eses servicios. Consideraciones finales: La evolución de las legislaciones relacionadas a la sangre y hemoderivados contribuyó en la consolidación de las atribuciones de los enfermeros en servicios de hemoterapia, pues garantizó el amparo legal y definió las prácticas a ser realizadas en eses servicios.

RESUMO Objetivo: Refletir sobre como a evolução das legislações ligadas a sangue e hemoderivados moldou a atuação do enfermeiro nos serviços de hemoterapia. Métodos: Estudo reflexivo, realizado mediante levantamentos em bases de dados, sobre a evolução da atuação do enfermeiro em serviços de hemoterapia. Resultados: Verificou-se que as diversas legislações, publicadas desde 1950, fomentaram a doação de sangue e moldaram a atuação do enfermeiro em serviços de hemoterapia, sendo as de maior relevância: a Lei nº 10.205/2001, sobre a coleta, processamento, estocagem, distribuição e aplicação do sangue; e a resolução do COFEN nº 629/2020, que aborda detalhadamente a atuação nesses serviços. Considerações finais: A evolução das legislações relacionadas a sangue e hemoderivados contribuiu na consolidação das atribuições dos enfermeiros em serviços de hemoterapia, pois garantiu o amparo legal e definiu as práticas a serem realizadas nesses serviços.