LentiPro stable producer cells: Delivering scalable and reliable lentiviral vector manufacturing

Lentiviral vectors are one of the most currently used viral vectors for gene and cell therapies. Their use in clinical protocols has significantly increased in the past 5 years with the approval of several gene therapeutic products relying on lentiviral vector gene delivery. Capable of transducing non-dividing cells and presenting safer integration profiles as self-inactivating vectors, lentiviral vectors have progressively undertaken gammaretroviral vector use in gene therapies. However the knowledge on lentiviral vector manufacture is far more immature than that of gammaretroviral vectors. While the production of gammaretrovirus rely on stable producer cell lines and perfusion systems, enabling high cell density and longer term productions, most of the bioprocesses for lentiviral bioproducts rely on transient transfections and short term batch productions. At the upstream process, many of the challenges lentiviral bioproducts present in their manufacturing are related to the apoptosis leading cytotoxicity of some of the vector components. Supported on our long track experience and enabling tools developed for gammaretrovirus manufacturing, we carried out the challenge of establishing a constitutive stable lentiviral producer cell line. To surpass the challenges we proposed to eliminate or reduce the cytotoxicity of the lentiviral vector expression components1. Several strategic novelties were introduced in the development of the cell line namely: (i) the use of a modified gag-pro-pol, (ii) introduction of all the third generation lentiviral expression cassettes by chemical transfection instead of viral transduction and (iii) performing only one clone screening step (enabling the use on the ‘Single step cloning screening’ protocol developed by our group2). After establishing a stable producer cell line the culture conditions were developed with the main aim of extending bioreaction culture time and viral vector total yields. A lentiviral producer cell line constitutively producing infective titers above 106 TU.mL-1.day-1 was established. Moreover the new protocol to generate the cell line enabled its development in less than six months. The cell line showed to be stable, consistently maintaining vector productivity over one month in the absence of antibiotics. At the bioreaction process it was possible to maintain the cells continuously producing over 10 days1. These results validate the transition to continuous or perfusion large-scale production systems qualifying the strengths and advantages of the strategies followed. This work to be presented will discuss the challenges on the manufacture and scale-up of lentiviral vectors as well the strategies and novel technologies to be adopted to enable effective upstream processes. 1Tomás et al. (2018) ‘LentiPro26: novel stable cell lines for constitutive lentiviral vector production\u27 Sci Rep. 8(1):5271 2 Rodrigues et al. (2015) ‘Single step cloning-screening method: a new tool for developing and studying high-titer viral vector producer cells\u27 Gene Ther. 22(9):6

Association between bilirubin and lipid profile in Portuguese elderly individuals

Several studies showed that mildly elevated bilirubin blood concentration is inversely associated to coronary artery disease (CAD). The most commonly mechanism contributing to CAD prevention is the bilirubin antioxidant effect, protecting several macromolecules from oxidation. Recently, other studies have found that higher serum bilirubin levels (BL) are associated with beneficial effects improving lipid profile and that might contribute to decrease cholesterol and triacylglycerol. In the meantime, a consistently negative association between BL and body mass index (BMI) was observed. The aim of this study was evaluate the cardiovascular risk protection by bilirubin analyzing the association between bilirubin levels and lipid profile in elderly. Clinical data, anthropometric measurements (BMI, Visceral Fat and Body Fat Percentage), lipid profile (total cholesterol-TC; triglycerides-TG; high-density lipoprotein cholesterol-HDL-c; low-density lipoprotein cholesterol-LDL-c; apoliprotein A-Apo-A; apoliprotein B-Apo-B) were evaluated in 70 institutionalized elderly, 43 females (mean age= 88.2±5.5 years old) and 27 males (mean age= 87.7±7.7 years old). Anthropometric measurements were obtained by bioimpedance, using specific scale (Tanita BC Model: 545). The lipid parameters were performed in autoanalyzer (Cobas Mira S, Roche, Switzerland) using available commercial kits. Correlation analysis revealed positive associations between BL and the HDL-c in males (total bilirubin: r=0.51; p<0.007) and no association in females (r=0.40; p<0.50). Positive associations were also found to TB levels and Apo-A (males: r=0.40; p<0.50 and females: r=0.270; p<0.013). Interestingly, the conjugated bilirubin present a stronger positive association with this two parameters (male: r=0.64; p<0.000; female: r=0.29; p<0.052). No associations were found between bilirubin and triglycerides; total cholesterol and anthropometric measurements. Low levels of HDL-c are a risk factor for coronary heart disease. A similar association has been shown for Apo-A1. These results suggest higher bilirubin is associated with higher cardioprotective agents, which indicates that the determination of total bilirubin should be included for more accurate cardiovascular disease risk assessment.info:eu-repo/semantics/publishedVersio

First experience of an undergraduate dental student with a reciprocating system in simulated root canals - a pilot study

Rotary instrumentation has been proposed in undergraduate teaching. The aim of this study was to evaluate student’s performance, through the obturation quality and treatment time, in a sequential range of L-simulated root canals. A senior undergraduate dental student sequentially prepared randomly numbered canals from 1 to 40, with the WaveOne Gold glider and primary file, according to the manufacturer instructions. A gutta-percha cone matched with the finishing instrument and epoxy resin-based sealer (AH Plus) was selected for the obturation. Three independent observers evaluated the obturation quality according to both density and length. Active, total instrumentation and obturation times were also measured. Statistical analysis was obtained by Mann–Whitney and Kruskal–Wallis tests with a significance level of p < 0.05. The quality of the obturation was independent of the number of prepared canals with adequate length and density in 87.5% of the prepared canals. Both active and total instrumentation, as well as obturation times, reduced significantly as the number of the prepared canals by the student increased (p < 0.05). The use of WaveOne Gold instrumentation and matched cone obturation by an inexperienced operator provided an adequate obturation quality in most of the curved simulated canals. The working time was significantly reduced through a short learning curve

Qualidade do sono e marcadores endócrinos e bioquímicos

O sono e o repouso constituem o ritmo biológico base da espécie humana e são fundamentais para uma boa saúde e qualidade de vida, com especial significado em crianças e jovens (DGS, 2015). Dormir bem é fundamental para a recuperação física e psíquica do indivíduo, indispensável para sermos saudáveis e essencial para nos mantermos ativos, concentrados e bem-dispostos. O sono é um equilibrador do humor e das emoções, recupera o corpo e a memória, estimula a criatividade e aumenta e consolida a capacidade de aprendizagem (Paiva & Penzel, 2011; SPN, 2015). Há cada vez mais evidências de que o sono tem influencia sobre os hábitos alimentares e consequentemente sobre o balanço energético e a regulação do peso corporal, estando também associado a outros fatores de risco metabólico na adolescência (Quist, Sjödin, Chaput & Hjorth, 2016). A má qualidade do sono tem um impacto negativo na qualidade de vida, verificando-se que a privação do sono é um fator de risco relevante para a saúde e segurança, aumentando o risco de doenças cardiovasculares, hipertensão, obesidade, diabetes, depressão, ansiedade e outros distúrbios de humor. A intervenção é fundamental, promovendo bons hábitos de sono desde a infância (Rebelo-Pinto, Pinto, Rebelo-Pinto & Paiva, 2014).info:eu-repo/semantics/publishedVersio

Dietary program impact on biochemical markers in diabetics: systematic review

Diabetes mellitus is a chronic disorder with major expansion worldwide. It’s estimated that the number of diabetes sufferers increase rapidly in the coming decades due to the population ageing (Ena,2016;IDF,2015). The nutrition intervention emphasizing the promotion of healthy eating has been shown to be an importante point in Diabetes Mellitus treatment since it fosters a better glycemic control and lipid profile (ADA2016)

Atividade física e impacto em marcadores bioquímicos em diabéticos: revisão sistemática da literatura

A diabetes mellitus é uma patologia crónica com grande expansão a nível mundial. Estima-se que o número de portadores de diabetes aumente rapidamente nas próximas décadas devido ao envelhecimento da população. Como tal, é fulcral consciencializar as pessoas para a adoção de estilos de vida mais ativos. A implementação de programas de intervenção com ênfase na promoção da atividade física tem demostrado ser um ponto importante no tratamento desta patologia permitindo um melhor controlo glicémico e perfil lipídico. Objetivos: Verificar qual a efetividade da implementação de um programa de atividade física nos valores de glicémia e perfil lipídico em portadores de diabetes mellitus. Metodologia: Revisão sistemática da literatura publicada nos anos 2010-2015 através da metodologia PICO na base de dados PubMed/Medline. Pretende-se responder à questão de investigação: “Qual a efetividade da implementação de um programa de atividade física nos valores de glicémia e perfil lipídico em indivíduos idosos portadores de diabetes mellitus?” Resultados: Após a aplicação dos critérios de inclusão foram selecionados 13 artigos de um total de 6600. Os artigos selecionados demostram que a implementação de programas de atividade física baseados em exercícios aeróbios, resistência, flexibilidade e combinados supervisionados, melhoram significativamente os níveis de glicémia e perfil lipídico. Conclusão: A implementação de programas de atividade física de acordo com informações cientificamente válidas, são pilares fundamentais no tratamento da diabetes mellitus, devendo ser fomentada a sua implementação.

Efeito de planos alimentares e de treino nos perfis glicémico e lipídico em idosos diabéticos tipo 2: revisão sistemática

A diabetes tipo 2 é uma patologia crónica com grande impacto a nível mundial particularmente na população idosa. Estima-se que o número de diabéticos tipo 2 aumente consideravelmente nas próximas décadas devido ao envelhecimento da população, obesidade, alimentação inadequada e inatividade física ou sedentarismo. A promoção de uma alimentação saudável e a prática de atividade física têm demonstrado ser fundamental no controlo e manutenção da glicémia e perfil lipídico. Objetivos: Verificar o impacto da implementação de programas dietéticos e/ou atividade física nos valores de glicémia e perfil lipídico em idosos diabéticos tipo 2. Métodos: Revisão sistemática da literatura publicada no período 2010-2016 na base de dados PubMed/Medline. Pretende-se responder à questão de investigação: “Qual o impacto da implementação de programas dietéticos e de atividade física nos valores de glicémia e perfil lipídico em idosos diabéticos?”. Após a aplicação dos critérios de inclusão foram selecionados 30 artigos de um total de 230825. Resultados: Nos 30 artigos selecionados são evidentes as modificações positivas que a implementação de programas dietéticos/atividade física baseados numa maior ingestão de produtos hortícolas, e alimentos com baixo índice glicémico, ácidos gordos insaturados (ómega 3 e 6), vitamina D, probióticos, restrição calórica bem como a prática de exercícios aeróbios, resistência e flexibilidade supervisionados têm no controlo da glicemia e perfil lipídico em idosos diabéticos tipo 2. Conclusões: A implementação de programas que visem uma alimentação saudável, equilibrada e a prática regular de atividade física de acordo com informações cientificamente válidas são pilares fundamentais no tratamento desta patologia

Efeito de planos alimentares e de treino nos perfis glicémico e lipidico em idosos diabéticos tipo 2: revisão sistemática

A diabetes tipo 2 é uma patologia crónica com grande impacto a nível mundial particularmente na população idosa. Estima-se que o número de diabéticos tipo 2 aumente consideravelmente nas próximas décadas devido ao envelhecimento da população, obesidade, alimentação inadequada e inatividade física. A promoção de uma alimentação saudável e a prática de atividade física têm demonstrado ser fundamentais no controlo e manutenção da glicémia e perfil lipídico. Objetivos: Verificar o impacto da implementação de programas dietéticos e/ou atividade física nos valores de glicémia e perfil lipídico em idosos diabéticos tipo 2. Métodos: Revisão sistemática da literatura publicada no período 2010-2016 na base de dados PubMed/Medline. Pretende-se responder à questão de investigação: “Qual o impacto da implementação de programas dietéticos e de atividade física nos valores de glicémia e perfil lipídico em idosos diabéticos?” Após a aplicação dos critérios de inclusão foram selecionados 30 artigos de um total de 230825. Resultados: Nos 30 artigos selecionados são evidentes as modificações positivas que a implementação de programas dietéticos/atividade física baseados numa maior ingestão de produtos hortícolas, alimentos com baixo índice glicémico, ácidos gordos insaturados, vitamina D, probióticos, restrição calórica bem como a prática de exercícios aeróbios, resistência e flexibilidade supervisionados têm no controlo da glicémia e perfil lipídico em idosos diabéticos tipo 2. Conclusões: A implementação de programas que visem uma alimentação saudável, equilibrada e a prática regular de atividade física de acordo com informações cientificamente válidas são pilares fundamentais no tratamento desta patologia.info:eu-repo/semantics/publishedVersio

Sleep quality and food intake of high school students

Poor sleep quality is associated with increased food intake and poor diet quality [1]. People with lack of sleep show a positive correlation. between free time and food intake and also experience hormonal and brain changes that drive the intake of food with high calorific value [1-3]. In addition, scientific research has shown a healthy and balanced diet to positively influence the quality of sleep [1]. Objective The present study was set out to assess the sleep quality of high school students in Bragança county, and its association with food intake. Methods The study used non-experimental, analytical and transversal methodology, of epidemiological character and with a quantitative approach. It was intended to carry out the study in a population of 862 high school students. However, due to consent being required from both legal guardians and students, a smaller sample of 345 students was obtained. The data was collected in May 2017 through a questionnaire that included the Pittsburgh Sleep Quality Index (PSQI), validated for the Portuguese population. Results Throughout the study and following PSQI analysis, it was concluded that 39.71% (n = 137) of participants showed poor quality of sleep (PSQI > 5 points). The correlation between sleep quality and food intake was assessed and a statistically significant association was found between the quality of sleep and the intake of snacks (X2 = 17.144; p = 0.000), sugary products (X2 = 18.603; p= 0 .000), fast-food (X2 = 12.353; p = 0.002) and ready meals (X2 = 14.852; p = 0.000). The risk of suffering from poor sleep quality is higher in young populations who frequently eat snacks ([OR]: 2.811; 99%), sugary products ([OR]: 1.901; 95%), fast-food ([OR]: 4.000; 99%) and ready meals ([OR]: 5.621; 95%) in comparison with young populations who rarely eat this sort of food. The sleep quality is also significantly related with the number of meals young people have in a day (X2 = 7.580; p = 0.023). The risk of having poor quality sleep is 2.240 times higher in young people who rarely eat 4-6 meals a day. Conclusions A correlation between sleep quality and food intake in the sampled students was seen. The risk of having poor quality of sleep is higher in students who frequently eat a high calorie diet and also in students who rarely have 4-6 meals a day. There are several connections between sleep quality and eating habits. Sleep promotion and its connection with standard diets should be included as an essential part of community empowerment for health-promoting lifestyles [1,4,5].info:eu-repo/semantics/publishedVersio

Physicochemical properties and bioaccessibility of phenolic compounds of dietary fibre concentrates from vegetable by-products

The agro-food industry generates a large volume of by-products, whose revaluation is essential for the circular economy. From these by-products, dietary fibre concentrates (DFCs) can be obtained. Therefore, the objective of this study was to characterise (a) the proximal composition by analysing soluble, insoluble and total Dietary Fibre (DF), (b) the physicochemical properties, and (c) the phenolic profile of artichoke, red pepper, carrot, and cucumber DFCs. In addition, the bioaccessibility of phenolic compounds was also evaluated after in vitro gastrointestinal and colonic digestions. The results showed that the DFCs had more than 30 g/100 g dw. The water holding and retention capacity of the DFCs ranges from 9.4 to 18.7 g of water/g. Artichoke DFC presented high concentration of phenolic compounds (8340.7 mg/kg) compared to the red pepper (304.4 mg/kg), carrot (217.4 mg/kg) and cucumber DFCs (195.7 mg/kg). During in vitro gastrointestinal digestion, soluble phenolic compounds were released from the food matrix, chlorogenic acid, the principal compound in artichoke and carrot DFCs, and hesperetin-7-rutinoside in red pepper cucumber DFCs. Total phenolic content decreased after in vitro colonic digestion hence the chemical transformation of the phenolic compounds by gut microbiota. Based on the results, DFCs could be good functional ingredients to develop DF-enriched food, reducing food waste.This research was funded by Ministerio de Ciencia y Tecnologia, Gobierno de Espana: AGLRTI-2018-094268-B-C21