ΔNp63 critical for progression of high grade non-muscle invasive bladder cancer through deregulation of specific genetic pathways

Tese de mestrado em Biologia Molecular e Genética, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2017Cancro da bexiga é o nono tumor mais comum a nível mundial, com uma estimativa de 429 000 novos casos registados, em 2012, e aproximadamente 165 000 mortes registadas, nesse mesmo ano, em todo o mundo. Os parâmetros clínicos e patológicos usados para avaliar o estádio do cancro da bexiga têm elevadas limitações e até à data ainda não existe nenhum biomarcador que preveja o comportamento tumoral em cada paciente. Por este motivo, é essencial a identificação de novos biomarcadores que estejam envolvidos na progressão tumoral e que permitam reconhecer diferentes tumores de acordo com a sua capacidade progressiva e invasiva. A classificação dos diferentes subtipos de tumor da bexiga permitirá desenvolver um tratamento especializado e individualizado, o que possibilitará uma melhoria da qualidade de vida e da taxa de sobrevivência destes doentes. O gene TP63 pertence à família do supressor de tumor TP53. No entanto ao contrário do TP53, que se encontra mutado em grande parte dos tumores, o gene TP63 raramente está na sua forma mutada. Ainda assim, a expressão do gene TP63 está alterada em alguns tumores. A isoforma do gene TP63, designada por ΔNp63, tem sido estudada como um possível biomarcador na área da oncologia, pois existem evidências que a relacionam com a progressão do tumor da bexiga não invasivo da camada muscular (NMIBC) para um tumor invasivo da camada muscular (MIBC). Estudos sugerem que esta isoforma inibe a senescência e a apoptose celular assim como promove a tumorogénese, proliferação e sobrevivência celular, exibindo características oncogénicas. No tecido normal da bexiga, o ΔNp63 tem uma alta/elevada expressão enquanto que no tecido tumoral de bexiga esta isoforma encontra-se subregulada. Este projeto foi iniciado em Mount Sinai, Nova Iorque, pela orientadora deste trabalho, Dr.ª Mireia Castillo, onde, após inativar o gene ΔNp63 das linhas celulares de NMIBC (RT112 e BFTC), observou que estas células apresentavam maior taxa de proliferação in vitro e in vivo, maior taxa de invasão in vitro e maior incidência de tumores primários e formação de metástases in vivo. Deste modo, foi necessário validar os resultados na Fundação Champalimaud e verificou-se, através do modelo ortotópico in vivo, que as células clones (BFTC_C e RT112_C) apresentaram maior agressividade tumoral e maior formação de metástases, assim confirmando o que tinha sido observado anteriormente. Através da análise da expressão génica, observou-se ainda que ambas as linhas celulares apresentavam nove e quatro genes, respetivamente sobrerregulados e subregulados, concomitantemente nas duas linhas celulares, respetivamente. Desta forma, o segundo objetivo deste projeto consistiu em descobrir quais dos genes selecionados na análise de expressão génica seriam mais relevantes na progressão tumoral. No entanto, apenas quatro dos nove sobrerregulados (AGR2, HOXC6, HRK e S100A4) e dois dos quatro genes subregulados (SCML1 e SPOCK1) foram estudados, com base no seu relacionamento com o desenvolvimento tumoral referido na literatura e também por existirem bons anticorpos e primers específicos. Deste modo, extraiu-se RNA das quatro linhas celulares (RT112_L_C, RT112_L_P, BFTC_L_C e BFTC_L_P) para realizar ensaios quantitativos de reação em cadeia de polimerase em tempo real (qRT-PCR) de forma a validar os resultados obtidos pela análise da expressão génica. A partir das mesmas linhas celulares foi extraída proteína para realizar ensaios de western blot (WB) e ensaios de imunofluorescência (IF), estes últimos foram realizados em células fixadas. Estas técnicas permitiram quantificar os níveis de proteína em cada linha celular (BFTC_P, BFTC_C, RT112_P e RT112_C). Após comparar os níveis de RNA mensageiro (mRNA) e de proteína, AGR2 foi o gene eleito como um dos favoritos a ter um papel essencial na progressão do tumor da bexiga. AGR2 tem maior expressão de mRNA nas células clones (ΔNp63-) comparativamente com as parentais (ΔNp63+) (observado apenas na linha celular designada BFTC). Relativamente à síntese de proteína, os resultados do WB sugerem que AGR2 é produzida em maior quantidade nas células clones (ΔNp63-) do que nas parentais (ΔNp63+) (em ambas as linhas celulares) e através de IF observa-se o mesmo resultado na linha celular BFTC. Outros estudos sugerem que o gene AGR2 tem funções extracelulares que aumentam a agressividade de diferentes tumores, como por exemplo o cancro do pulmão, da mama e da próstata. Além disso, expressão de mRNA de AGR2 é elevada no carcinoma de células de transição da bexiga, adenocarcinoma da próstata e do pulmão. A desativação do gene foi feita em cinco conjuntos de células de cada linha celular de NMIBC, onde já tinha sido inativado a isoforma ΔNp63 do gene TP63, formando cinco clones de cada linha celular, designados por BFTC_A1, BFTC_A2, BFTC_A3, BFTC_A4 e BFTC_A5, assim como para a linha celular RT112. Após silenciamento do gene AGR2 por shRNA nas células BFTC_C e RT112_C, estudaram-se os níveis de mRNA por qRT-PCR, e de proteína através das técnicas de WB e IF para determinar a eficiência da eliminação do mesmo. Estes estudos só foram realizados para a linha celular BFTC. O clone BFTC_A5 mostrou ser o que apresentava menor nível de mRNA e respetiva proteína AGR2, por isso esta linha celular foi utilizada para estudos funcionais, como ensaios de ciclo celular, ensaios de proliferação e ensaios de invasão celular. Os estudos funcionais foram essenciais para observar se o gene AGR2 está envolvido na progressão celular. Ensaios de ciclo celular revelaram que as células tumorais designadas BFTC_A5 (ΔNp63- AGR2-) apresentaram menor divisão celular comparativamente às células tumorais BFTC_C (ΔNp63- AGR2+) e BFTC_P (ΔNp63+ AGR2+), in vitro. As células tumorais BFTC_A5 (ΔNp63- AGR2-) revelaram ainda menor capacidade de proliferação e de invasão celular, in vitro. Estes estudos sugerem que o gene AGR2 pode estar envolvido na progressão do tumor da bexiga não invasivo da camada muscular (NMIBC) para invasivo da camada muscular (MIBC). As células tumorais que sintetizam em simultâneo a isoforma ΔNp63 e o gene AGR2 (BFTC_P) revelaram não ter capacidades tão proliferativas e tão invasoras como as células tumorais que apenas sintetizam AGR2 (BFTC_C). No entanto quando o gene AGR2 é desligado, estas células tumorais voltam a perder a sua capacidade invasora e proliferativa, sugerindo assim que o gene AGR2 é essencial para a proliferação e invasão tumoral. Este trabalho é o primeiro que estuda a interação entre o produto do gene AGR2 e a isoforma ΔNp63 do gene TP63 na progressão do tumor da bexiga não invasivo da camada muscular. No entanto mais estudos serão necessários para confirmar esta hipótese, nomeadamente estudos com o modelo ortotópico in vivo e também a análise de imunofluorescência de microarrays de tumores (TMA) que irá validar os resultados in vitro, assim como correlacionar a expressão do AGR2 com as características do doente (prognóstico, presença de metástases, risco de progressão, entre outros). Além disso, é também necessário realizar os mesmos ensaios, descritos neste trabalho, para a linha celular RT112. Neste trabalho, AGR2 é descrito como um potencial fator de progressão de NMIBC para MIBC. O gene AGR2 é importante para o desenvolvimento do tumor e poderá ser um biomarcador com a capacidade de distinguir tumores que devem sofrer um tratamento mais ou menos invasivo. Como por exemplo, pacientes com tumores de elevado risco de progressão poderão beneficiar de uma cistectomia, enquanto que pacientes com tumores de baixo risco de progressão poderão usufruir de um tratamento menos invasivo e de vigilância médica com alguma regularidade. A existência de um anticorpo contra a proteína AGR2 pode permitir o desenvolvimento de ensaios clínicos em doentes com NMIBC, de modo a testar a sua efetividade no controlo da progressão destes mesmos tumores.ΔNp63, an isoform of TP63 gene, has been described as an essential marker for Non-Muscle Invasive Bladder Cancer (NMIBC) progression. After knocking down ΔNp63 in NMIBC cell lines, nine and four genes were found up-regulated and down-regulated, respectively, through gene expression analysis, in two different bladder tumor cell lines (RT112 and BFTC). From this set of genes, four up-regulated (AGR2, HOXC6, HRK and S100A4) and two down-regulated genes (SCML1 and SPOCK1) were chosen for further analysis due to the availability of good antibodies and primers, in order to discover which of those genes are more relevant to bladder cancer progression. With this purpose, total RNA was extracted from BFTC and RT112 cells (both parental and knock down for ΔNp63) to perform quantitative real-time polymerase chain reaction (qRT-PCR) and to validate the gene expression analysis. From the same cell lines, proteins were extracted to performed western blot (WB) analysis and immunofluorescence (IF) assays and the protein and RNA levels were compared. The obtained results suggest that AGR2 gene was the most related in bladder cancer progression. In addition, mRNA level of AGR2 was higher in knock down clones than in parental cells (verified only in BFTC cells) and WB results showed that AGR2 protein was also higher in knock down than in parental cells. Similar results were observed by IF assay in BFTC cells. Other studies suggested that AGR2 has extracellular functions that increase aggressiveness of different tumors. Moreover, AGR2 mRNA expression has been described as high in urinary bladder transitional cell carcinoma, prostate adenocarcinoma and lung carcinoma. To study the effects of that gene in bladder tumor progression, AGR2 was knocked down in RT112_C and BFTC_C cells which had already been knocked down for ΔNp63 and mRNA and protein level studies were performed again through qRT-PCR, WB and IF techniques. After selection of the cell line with lower level of mRNA and protein AGR2, BFTC_A5, functional studies were performed, such as cell cycle, invasion and proliferation assays. Functional studies showed that BFTC_A5 (ΔNp63- AGR2-) cells displayed lower cell division, lower proliferative and invasive capacities compared with BFTC_C (ΔNp63- AGR2+), which suggest that AGR2 may has a role in bladder tumor progression

Light microclimate-driven changes at transcriptional level in photosynthetic grape berry tissues

Viticulture practices that change the light distribution in the grapevine canopy can interfere with several physiological mechanisms, such as grape berry photosynthesis and other metabolic pathways, and consequently impact the berry biochemical composition, which is key to the final wine quality. We previously showed that the photosynthetic activity of exocarp and seed tissues from a white cultivar (Alvarinho) was in fact responsive to the light microclimate in the canopy (low and high light, LL and HL, respectively), and that these different light microclimates also led to distinct metabolite profiles, suggesting a berry tissue-specific interlink between photosynthesis and metabolism. In the present work, we analyzed the transcript levels of key genes in exocarps and seed integuments of berries from the same cultivar collected from HL and LL microclimates at three developmental stages, using real-time qPCR. In exocarp, the expression levels of genes involved in carbohydrate metabolism (VvSuSy1), phenylpropanoid (VvPAL1), stilbenoid (VvSTS1), and flavan-3-ol synthesis (VvDFR, VvLAR2, and VvANR) were highest at the green stage. In seeds, the expression of several genes associated with both phenylpropanoid (VvCHS1 and VvCHS3) and flavan-3-ol synthesis (VvDFR and VvLAR2) showed a peak at the véraison stage, whereas that of RuBisCO was maintained up to the mature stage. Overall, the HL microclimate, compared to that of LL, resulted in a higher expression of genes encoding elements associated with both photosynthesis (VvChlSyn and VvRuBisCO), carbohydrate metabolism (VvSPS1), and photoprotection (carotenoid pathways genes) in both tissues. HL also induced the expression of the VvFLS1 gene, which was translated into a higher activity of the FLS enzyme producing flavonol-type flavonoids, whereas the expression of several other flavonoid pathway genes (e.g., VvCHS3, VvSTS1, VvDFR, and VvLDOX) was reduced, suggesting a specific role of flavonols in photoprotection of berries growing in the HL microclimate. This work suggests a possible link at the transcriptional level between berry photosynthesis and pathways of primary and secondary metabolism, and provides relevant information for improving the management of the light microenvironment at canopy level of the grapes.This research and the APC was funded by FCT—Portuguese Foundation for Science and Technology, grant provided to Andreia Garrido (PD/BD/128275/2017), under the Doctoral Programme “Agricultural Production Chains—from fork to farm” (PD/00122/2012), and by the European Social Funds and the Regional Operational Programme Norte 2020.info:eu-repo/semantics/publishedVersio

Metabolomics of photosynthetically active tissues in white grapes: effects of light microclimate and stress mitigation strategies

The effects of climate change are becoming a real concern for the viticulture sector, with impacts on both grapevine physiology and the quality of the fresh berries and wine. Short-term mitigation strategies, like foliar kaolin application and smart irrigation regimes, have been implemented to overcome these problems. We previously showed that these strategies also influence the photosynthetic activity of the berries themselves, specifically in the exocarp and seed. In the present work, we assessed the modulating effects of both canopy-light microclimate, kaolin and irrigation treatments on the metabolic profiles of the exocarp and seed, as well as the potential role of berry photosynthesis herein. Berries from the white variety Alvarinho were collected at two contrasting light microclimate positions within the vine canopy (HLhigh light and LLlow light) from both irrigated and kaolin-treated plants, and their respective controls, at three fruit developmental stages (green, véraison and mature). Untargeted liquid chromatography mass spectrometry (LCMS) profiling of semi-polar extracts followed by multivariate statistical analysis indicate that both the light microclimate and irrigation influenced the level of a series of phenolic compounds, depending on the ripening stage of the berries. Moreover, untargeted gas chromatography mass spectrometry (GCMS) profiling of polar extracts show that amino acid and sugar levels were influenced mainly by the interaction of irrigation and kaolin treatments. The results reveal that both photosynthetically active berry tissues had a distinct metabolic profile in response to the local light microclimate, which suggests a specific role of photosynthesis in these tissues. A higher light intensity within the canopy mainly increased the supply of carbon precursors to the phenylpropanoid/flavonoid pathway, resulting in increased levels of phenolic compounds in the exocarp, while in seeds, light mostly influenced compounds related to carbon storage and seed development. In addition, our work provides new insights into the influence of abiotic stress mitigation strategies on the composition of exocarps and seeds, which are both important tissues for the quality of grape-derived products.This research and APC were funded by the FCT-Portuguese Foundation for Science and Technology, grant provided to Andreia Garrido (PD/BD/128275/2017), under the Doctoral Program "Agricultural Production Chains-from fork to farm" (PD/00122/2012).info:eu-repo/semantics/publishedVersio

Fruit photosynthesis: more to know about where, how and why

Not only leaves but also other plant organs and structures typically considered as carbon sinks, including stems, roots, flowers, fruits and seeds, may exhibit photosynthetic activity. There is still a lack of a coherent and systematized body of knowledge and consensus on the role(s) of photosynthesis in these “sink” organs. With regard to fruits, their actual photosynthetic activity is influenced by a range of properties, including fruit anatomy, histology, physiology, development and the surrounding microclimate. At early stages of development fruits generally contain high levels of chlorophylls, a high density of functional stomata and thin cuticles. While some plant species retain functional chloroplasts in their fruits upon subsequent development or ripening, most species undergo a disintegration of the fruit chloroplast grana and reduction in stomata functionality, thus limiting gas exchange. In addition, the increase in fruit volume hinders light penetration and access to CO2, also reducing photosynthetic activity. This review aimed to compile information on aspects related to fruit photosynthesis, from fruit characteristics to ecological drivers, and to address the following challenging biological questions: why does a fruit show photosynthetic activity and what could be its functions? Overall, there is a body of evidence to support the hypothesis that photosynthesis in fruits is key to locally providing: ATP and NADPH, which are both fundamental for several demanding biosynthetic pathways (e.g., synthesis of fatty acids); O2, to prevent hypoxia in its inner tissues including seeds; and carbon skeletons, which can fuel the biosynthesis of primary and secondary metabolites important for the growth of fruits and for spreading, survival and germination of their seed (e.g., sugars, flavonoids, tannins, lipids). At the same time, both primary and secondary metabolites present in fruits and seeds are key to human life, for instance as sources for nutrition, bioactives, oils and other economically important compounds or components. Understanding the functions of photosynthesis in fruits is pivotal to crop management, providing a rationale for manipulating microenvironmental conditions and the expression of key photosynthetic genes, which may help growers or breeders to optimize development, composition, yield or other economically important fruit quality aspects.This research was funded by FCT—Portuguese Foundation for Science and Technology, via a grant provided to AG (PD/BD/128275/2017), under the Doctoral Programme “Agricultural Production Chains—from fork to farm” (PD/00122/2012), and by the European Social Funds and the Regional Operational Programme Norte 2020. FCT also supported the work via CITAB and CBMA research units under the projects UIDB/04033/2020 and UIDB/04050/2020, respectively. The work was also support by project I&D&I “AgriFood XXI”, ref. NORTE-01-0145-FEDER-000041, co-financed by the European Regional Development Fund (FEDER), through NORTE 2020 (Northern Regional Operational Program 2014/2020). This work was also supported by CESAM (UIDP/50017/2020 + UIDB/50017/2020 + LA/P/0094/2020). Ric C. H. De Vos was financed by the business unit Bioscience of Wageningen University and Research Centre

Effects of Sitagliptin Treatment on Dysmetabolism, Inflammation, and Oxidative Stress in an Animal Model of Type 2 Diabetes (ZDF Rat)

The purpose of this paper is to evaluate the chronic effect of sitagliptin on metabolic profile, inflammation, and redox status in the Zucker Diabetic Fatty (ZDF) rat, an animal model of obese type 2 diabetes. Diabetic and obese ZDF (fa/fa) rats and their controls (ZDF +/+) were treated during 6 weeks with vehicle (control) and sitagliptin (10mg/kg/bw). Glucose, HbA1c, insulin, Total-c, TGs, IL-1β, TNF-α, CRPhs, and adiponectin were assessed in serum and MDA and TAS in serum, pancreas, and heart. Pancreatic histology was also evaluated. Sitagliptin in diabetic rats promoted a decrease in glucose, HbA1c, Total-c, and TGs accompanied by a partial prevention of insulinopenia, together, with a decrease in CRPhs and IL-1β. Sitagliptin also showed a positive impact on lipid peroxidation and hypertension prevention. In conclusion, chronic sitagliptin treatment corrected the glycaemic dysmetabolism, hypertriglyceridaemia, inflammation, and hypertension, reduced the severity of the histopathological lesions of pancreatic endocrine and exocrine tissues, together with a favourable redox status, which might be a further advantage in the management of diabetes and its proatherogenic comorbidities

Effects of Sitagliptin Treatment on Dysmetabolism, Inflammation, and Oxidative Stress in an Animal Model of Type 2 Diabetes (ZDF Rat)

The purpose of this paper is to evaluate the chronic effect of sitagliptin on metabolic profile, inflammation, and redox status in the Zucker Diabetic Fatty (ZDF) rat, an animal model of obese type 2 diabetes. Diabetic and obese ZDF (fa/fa) rats and their controls (ZDF +/+) were treated during 6 weeks with vehicle (control) and sitagliptin (10 mg/kg/bw). Glucose, HbA1c, insulin, Total-c, TGs, IL-1β, TNF-α, CRPhs, and adiponectin were assessed in serum and MDA and TAS in serum, pancreas, and heart. Pancreatic histology was also evaluated. Sitagliptin in diabetic rats promoted a decrease in glucose, HbA1c, Total-c, and TGs accompanied by a partial prevention of insulinopenia, together, with a decrease in CRPhs and IL-1β. Sitagliptin also showed a positive impact on lipid peroxidation and hypertension prevention. In conclusion, chronic sitagliptin treatment corrected the glycaemic dysmetabolism, hypertriglyceridaemia, inflammation, and hypertension, reduced the severity of the histopathological lesions of pancreatic endocrine and exocrine tissues, together with a favourable redox status, which might be a further advantage in the management of diabetes and its proatherogenic comorbidities

EBV and MSI status in gastric cancer: does it matter?

We investigated the impactof microsatellite instability (MSI) and Epstein–Barr virus (EBV) status in gastric cancer (GC), regarding response to perioperative chemotherapy (POPChT), overall survival (OS), and progression-free survival (PFS). We included 137 cases of operated GC, 51 of which were submitted to POPChT. MSI status was determined by multiplex PCR and EBV status by EBV-encoded RNA in situ hybridization. Thirty-seven (27%) cases presented as MSI-high, and seven (5.1%) were EBV+. Concerning tumor regression after POPChT, no differences were observed between the molecular subtypes, but females were more likely to respond (p = 0.062). No significant differences were found in OS or PFS between different subtypes. In multivariate analysis, age (HR 1.02, IC 95% 1.002–1.056, p = 0.033) and positive lymph nodes (HR 1.82, IC 95% 1.034–3.211, p = 0.038) were the only prognostic factors for OS. However, females with MSI-high tumors treated with POPChT demonstrated a significantly increased OS compared to females with MSS tumors (p = 0.031). In conclusion, we found a high proportion of MSI-high cases. MSI and EBV status did not influence OS or PFS either in patients submitted to POPChT or surgery alone. However, superior survival of females with MSI-high tumors suggests that sex disparities and molecular classification may influence treatment options in GC.info:eu-repo/semantics/publishedVersio

Does It Matter?

Funding Information: This research was partially co-financed by Hospital da Luz Lisboa under the initiative “Luz Investigação” in the context of the Group GENIUS (Reference LH.INV.F2019015).We investigated the impactof microsatellite instability (MSI) and Epstein–Barr virus (EBV) status in gastric cancer (GC), regarding response to perioperative chemotherapy (POPChT), overall survival (OS), and progression-free survival (PFS). We included 137 cases of operated GC, 51 of which were submitted to POPChT. MSI status was determined by multiplex PCR and EBV status by EBV-encoded RNA in situ hybridization. Thirty-seven (27%) cases presented as MSI-high, and seven (5.1%) were EBV+. Concerning tumor regression after POPChT, no differences were observed between the molecular subtypes, but females were more likely to respond (p = 0.062). No significant differences were found in OS or PFS between different subtypes. In multivariate analysis, age (HR 1.02, IC 95% 1.002–1.056, p = 0.033) and positive lymph nodes (HR 1.82, IC 95% 1.034–3.211, p = 0.038) were the only prognostic factors for OS. However, females with MSI-high tumors treated with POPChT demonstrated a significantly increased OS compared to females with MSS tumors (p = 0.031). In conclusion, we found a high proportion of MSI-high cases. MSI and EBV status did not influence OS or PFS either in patients submitted to POPChT or surgery alone. However, superior survival of females with MSI-high tumors suggests that sex disparities and molecular classification may influence treatment options in GC.publishersversionpublishe

Política Nacional de Saúde Bucal: fatores associados à integralidade do cuidado

OBJECTIVE: To analyze the factors associated with comprehensiveness in oral health care in Centers of Dental Specialists, according to the guiding principles of the Brazilian Oral Health Policy. METHODS: An exploratory cross-sectional study, based on an interview with 611 users of four specialized dental care centers, was performed in the state of Bahia, Northeastern Brazil, in 2008. The dependent variable was described as "comprehensiveness in oral health care", corresponding to having a primary dental care performed before specialized treatment or concomitantly with it. The main covariables referred to the level of coverage of the family health strategy in the city, users' sociodemographic characteristics, and organizational and geographic accessibility to the service, in addition to the type of specialized care required. RESULTS: Residents of the cities where the Family Healthcare Program had a coverage >;50% were more likely to conclude their dental treatment (PR=2.03, 95% CI: 1.33;3.09), compared to those who lived in places with lower coverage. Individuals who sought endodontic treatment were more likely to receive comprehensive oral health care than users who were seeking other types of specialized care (PR=2.31, 95% CI: 1.67;3.19). Users with better geographic accessibility to specialized services (PR=1.22, 95% CI: 1.03;1.41), with a reference guide from primary care (PR=2.95, 95% CI: 1.82;4.78) and coming from primary health care services (PR=3.13, 95% CI: 1.70;5.77) were more likely to achieve comprehensiveness in oral health care than other users. CONCLUSIONS: Users with better geographic accessibility, lower age and need of endodontic services were more likely to receive comprehensive health care. Implementation of Centers of Dental Specialists in cities where primary healthcare is not adequately structured is not recommended, because secondary health care would meet the free demand and perform basic procedures, thus not fulfilling the expected principle of comprehensiveness.OBJETIVO: Analizar factores relacionados a la integración en la asistencia a la salud bucal en centros de especialidades odontológicas según los principios que guían la Política Nacional de Salud Bucal. MÉTODOS: Estudio exploratorio transversal basado en entrevista con 611 usuarios de cuatro centros de especialidades odontológicas de Bahia, Noreste de Brasil, en 2008. La variable dependiente fue descrita como "integración en la salud bucal", correspondiente a la realización de tratamiento odontológico básico antes del tratamiento especializado o concomitante a este. Las principales covariables se refirieron a cobertura de la estrategia salud de la familia en el municipio, características sociodemográficas de los usuarios, accesibilidad organizacional y geográfica al servicio, además del tipo de especialidad demandada. RESULTADOS: Residentes de ciudades donde el Programa Salud de la Familia tenía cobertura >;50% tuvieron más chance de concluir el tratamiento odontológico (RP=2,03, IC 95%: 1,33;3,09) con relación a aquellas que estaban residenciados en localidades con cobertura menor. Quien buscó tratamiento endodóntico tuvo más chance de recibir asistencia integral a la salud bucal en comparación con los usuarios que buscan otras especialidades (RP=2,31, IC 95%: 1,67;3,19). Los usuarios con mayor facilidad en el acceso geográfico al servicio especializado (RP=1,22, IC 95%: 1,03;1,41), con ficha de referencia (RP=2,95, IC 95%: 1,82;4,78) y oriundos de la atención primaria (RP=3,13, IC 95%: 1,70;5,77) tuvieron más chance de alcanzar la integración en la asistencia a la salud bucal con relación a los demás usuarios. CONCLUSIONES: Usuarios con facilidad de acceso geográfico, más jóvenes y necesidad de servicios endodóntico tuvieron más chance de recibir asistencia integral. La implantación de centros de especialidades odontológicas en municipios donde la atención primaria a la salud no se encuentre adecuadamente estructurada no es recomendada, visto que la atención secundaria estaría atendiendo la libre demanda y ejecutando procedimientos básicos y, por lo tanto, no cumpliendo el principio de la integración pretendida.OBJETIVO: Analisar fatores relacionados à integralidade na assistência à saúde bucal em centros de especialidades odontológicas segundo os princípios norteadores da Política Nacional de Saúde Bucal. MÉTODOS: Estudo exploratório transversal baseado em entrevista com 611 usuários de quatro centros de especialidades odontológicas da Bahia em 2008. A variável dependente foi descrita como "integralidade na saúde bucal", correspondente à realização de tratamento odontológico básico antes do tratamento especializado ou concomitante a este. As principais co-variáveis se referiram a cobertura da estratégia saúde da família no município, características sociodemográficas dos usuários, acessibilidade organizacional e geográfica ao serviço, além do tipo de especialidade demandada. RESULTADOS: Residentes de cidades em que o Programa Saúde da Família tinha cobertura >; 50% tiveram mais chance de concluir o tratamento odontológico (RP = 2,03, IC 95%: 1,33;3,09) em relação àqueles residentes em locais com cobertura menor. Quem buscou tratamento endodôntico teve mais chance de receber assistência integral à saúde bucal do que os usuários em busca de outras especialidades (RP = 2,31, IC 95%: 1,67;3,19). Os usuários com maior facilidade no acesso geográfico ao serviço especializado (RP = 1,22, IC 95%: 1,03;1,41), com ficha de referência (RP = 2,95, IC 95%: 1,82;4,78) e oriundos da atenção primária (RP = 3,13, IC 95%: 1,70;5,77) tiveram mais chance de alcançar a integralidade na assistência à saúde bucal em relação aos demais usuários. CONCLUSÕES: Usuários com facilidade de acesso geográfico, mais jovens e necessidade de serviço endodôntico tiveram mais chance de receber assistência integral. A implantação de centros de especialidades odontológicas em municípios nos quais a atenção primária à saúde não esteja adequadamente estruturada não é recomendada, visto que a atenção secundária estaria atendendo a livre demanda e executando procedimentos básicos e, portanto, não cumprindo o princípio da integralidade pretendida

Modelling gas-liquid mass transfer in wastewater treatment : when current knowledge needs to encounter engineering practice and vice versa

Abstract Gas–liquid mass transfer in wastewater treatment processes has received considerable attention over the last decades from both academia and industry. Indeed, improvements in modelling gas–liquid mass transfer can bring huge benefits in terms of reaction rates, plant energy expenditure, acid–base equilibria and greenhouse gas emissions. Despite these efforts, there is still no universally valid correlation between the design and operating parameters of a wastewater treatment plant and the gas–liquid mass transfer coefficients. That is why the current practice for oxygen mass transfer modelling is to apply overly simplified models, which come with multiple assumptions that are not valid for most applications. To deal with these complexities, correction factors were introduced over time. The most uncertain of them is the α-factor. To build fundamental gas–liquid mass transfer knowledge more advanced modelling paradigms have been applied more recently. Yet these come with a high level of complexity making them impractical for rapid process design and optimisation in an industrial setting. However, the knowledge gained from these more advanced models can help in improving the way the α-factor and thus gas–liquid mass transfer coefficient should be applied. That is why the presented work aims at clarifying the current state-of-the-art in gas–liquid mass transfer modelling of oxygen and other gases, but also to direct academic research efforts towards the needs of the industrial practitioners