Respostas imunitárias em Bathymodiolus azoricus desafiados com Vibrio : abordagens para caracterizar as estratégias de sobrevivência do mexilhão e suas adaptações fisiológicas em ambientes extremos do mar profundo

Tese de Doutoramento, Ciências do Mar, especialidade de Biologia Marinha, 18 de Dezembro de 2015, Universidade dos Açores.O ambiente marinho profundo representa uma excelente fonte de investigação para desenvolver e melhorar os conhecimentos sobre os animais que habitam em ambientes extremos e sobre as relações quimiossintéticas estabelecidas nesse ecossistema marinho. Dessa premissa surge a necessidade de entender o funcionamento do sistema imunitário em animais do fundo marinho, nomeadamente do mexilhão do mar profundo Bathymodiolus azoricus. O facto de este mexilhão habitar e desenvolver-se em ambientes extremos, como as fontes hidrotermais marinhas de profundidade, torna-o num interessante modelo para caracterizar as suas estratégias de sobrevivência e adaptações fisiológicas nesses ambientes. Nestas fontes hidrotermais, o seu sistema imunitário está constantemente a ser desafiado por compostos estranhos e microrganismos, como as bactérias do meio ambiente circundante. Contudo, as respostas imunitárias no B. azoricus após contacto com Vibrio não estão completamente compreendidas. As bactérias do género Vibrio são frequentemente patogénicos marinhos que causam grande mortalidade em bivalves. Assim, a presente tese pretende compreender as respostas imunitárias dos mexilhões após a exposição a diferentes estirpes de Vibrio, de modo a elucidar as principais vias de sinalização dos genes e proteínas intervenientes na defesa dos mexilhões durante um determinado estímulo bacteriano. Neste sentido, os métodos apresentados nesta tese têm como base a utilização de diferentes técnicas da biologia molecular, com destaque para o PCR quantitativo em tempo real baseado no princípio da reacção em cadeia da polimerase. Esta técnica foi realizada: em brânquia de mexilhões B. azoricus submetidos ao contacto com diferentes suspensões bacterianas de patogénicos marinhos (Capítulos I); no estudo comparativo da expressão de genes e proteínas entre o mexilhão de profundidade B. azoricus e o mexilhão costeiro Mytilus galloprovincialis (Capitulo II) e no estudo sobre a especificidade da resposta imunitária em diferentes tecidos, entre populações de B. azoricus provenientes das fontes hidrotermais Menez Gwen e Lucky Strike, após contacto com a bactéria Vibrio diabolicus (Capitulo III). [...].ABSTRACT: The deep-sea environment is excellent source for investigating and improving our knowledge in the field of marine biology of organisms living in extreme habitats and their relationships with chemosynthetic-based ecosystems. Along with the study of chemosynthetic-based ecosystems the necessity to understand immune system function in deep-sea hydrothermal animals such as in the mussel Bathymodiolus azoricus, has emerged gradually. Indeed, these mussels live and thrive in extreme environments as the deep-sea hydrothermal vents. B. azoricus species has become an interesting model to characterize the survival strategies and physiological adaptations in these environments. At such depths and under hydrothermal physico-chemical conditions, their immune system is constantly challenged by foreign vent compounds and microorganisms (e.g. bacteria). However, these immune responses in B. azoricus upon Vibrio challenges are not thoroughly understood. Vibrio bacteria are frequently pathogens in the aquatic environment that cause a high mortality in bivalves. This thesis was conceived and achieved with the aim of better understanding the immune response in B. azoricus mussels after several experimental challenges with different Vibrio strains. The core of this thesis consisted of elucidating signaling gene pathways and proteins involved in mussel defenses during bacterial stimuli. In this way, the thesis methods were undertaken using several molecular biology techniques, highlighting the qPCR. This technique was used: in gills from B. azoricus mussels infected with suspension of different marine bacterial pathogens (Chapter I); in comparative studies of gene expression and protein analyses between deep-sea hydrothermal vent mussel B. azoricus and the shallow-water mussel Mytilus galloprovincialis (Chapter II) and in an immune response tissue-specificity study in different tissues, and between Menez Gwen and Lucky Strike B. azoricus populations, after exposure to Vibrio diabolicus bacterium (Chapter III). [...]This thesis was supported by FCT grant SFRH/BD/68951/2010, Department of Oceanography and Fisheries, University of the Azores and Institute of Marine Research (IMAR)

Characterisation of microbial attack on archaeological bone

As part of an EU funded project to investigate the factors influencing bone preservation in the archaeological record, more than 250 bones from 41 archaeological sites in five countries spanning four climatic regions were studied for diagenetic alteration. Sites were selected to cover a range of environmental conditions and archaeological contexts. Microscopic and physical (mercury intrusion porosimetry) analyses of these bones revealed that the majority (68%) had suffered microbial attack. Furthermore, significant differences were found between animal and human bone in both the state of preservation and the type of microbial attack present. These differences in preservation might result from differences in early taphonomy of the bones. © 2003 Elsevier Science Ltd. All rights reserved

Global variation in postoperative mortality and complications after cancer surgery: a multicentre, prospective cohort study in 82 countries

© 2021 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY-NC-ND 4.0 licenseBackground: 80% of individuals with cancer will require a surgical procedure, yet little comparative data exist on early outcomes in low-income and middle-income countries (LMICs). We compared postoperative outcomes in breast, colorectal, and gastric cancer surgery in hospitals worldwide, focusing on the effect of disease stage and complications on postoperative mortality. Methods: This was a multicentre, international prospective cohort study of consecutive adult patients undergoing surgery for primary breast, colorectal, or gastric cancer requiring a skin incision done under general or neuraxial anaesthesia. The primary outcome was death or major complication within 30 days of surgery. Multilevel logistic regression determined relationships within three-level nested models of patients within hospitals and countries. Hospital-level infrastructure effects were explored with three-way mediation analyses. This study was registered with ClinicalTrials.gov, NCT03471494. Findings: Between April 1, 2018, and Jan 31, 2019, we enrolled 15 958 patients from 428 hospitals in 82 countries (high income 9106 patients, 31 countries; upper-middle income 2721 patients, 23 countries; or lower-middle income 4131 patients, 28 countries). Patients in LMICs presented with more advanced disease compared with patients in high-income countries. 30-day mortality was higher for gastric cancer in low-income or lower-middle-income countries (adjusted odds ratio 3·72, 95% CI 1·70–8·16) and for colorectal cancer in low-income or lower-middle-income countries (4·59, 2·39–8·80) and upper-middle-income countries (2·06, 1·11–3·83). No difference in 30-day mortality was seen in breast cancer. The proportion of patients who died after a major complication was greatest in low-income or lower-middle-income countries (6·15, 3·26–11·59) and upper-middle-income countries (3·89, 2·08–7·29). Postoperative death after complications was partly explained by patient factors (60%) and partly by hospital or country (40%). The absence of consistently available postoperative care facilities was associated with seven to 10 more deaths per 100 major complications in LMICs. Cancer stage alone explained little of the early variation in mortality or postoperative complications. Interpretation: Higher levels of mortality after cancer surgery in LMICs was not fully explained by later presentation of disease. The capacity to rescue patients from surgical complications is a tangible opportunity for meaningful intervention. Early death after cancer surgery might be reduced by policies focusing on strengthening perioperative care systems to detect and intervene in common complications. Funding: National Institute for Health Research Global Health Research Unit

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

© 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 licenseBackground: Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide. Methods: A multimethods analysis was performed as part of the GlobalSurg 3 study—a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital. Findings: Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3·85 [95% CI 2·58–5·75]; p<0·0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63·0% vs 82·7%; OR 0·35 [0·23–0·53]; p<0·0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer. Interpretation: Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised. Funding: National Institute for Health and Care Research