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- Minicircular DNA as a potential gene therapy vector in cervical cancerPublication . Alves, Joel Marques; Sousa, Ângela Maria Almeida de; Sousa, Fani Pereira deThe Human Papillomavirus (HPV) infects a large number of people worldwide and is associated with more than 99% of cervical cancer, being a second major cause of cancer death in women. HPV E6 oncoprotein is responsible for the progression of this tumor due to the inactivation of the p53 tumor suppressor. Thus, novel therapeutic approaches for the treatment of this cancer have been explored, for example gene therapy, which is based on the insertion of genetic material of interest into the cells, resulting in the expression of proteins encoded by the respective genes. Plasmid DNA has been the non-viral vector mainly used in DNA therapeutics due to its simple manufacture, low toxicity and immunogenicity. Despite some successful cases, the possibility of using a DNA vector of reduced size, decreased toxicity by elimination of the bacterial cassette and able to favor the target gene expression by increased transfection efficiency, makes the minicircular DNA (mcDNA) a very promising gene therapy vector. Therefore, the present work aims to construct a mcDNA vector encoding p53 to reestablish the levels of this tumor suppressor protein in cervical cancer cells and to induce apoptosis. The production of mcDNA started by cloning the p53-encoding sequence into a parental plasmid (PP). Then, Escherichia coli ZYCY10P3S2T strain was transformed by heat shock with the p53-PP vector. The use of this E. coli strain is of major importance since it allows the plasmid recombination to form mcDNA and miniplasmid species by induction with L-arabinose during cell growth. Moreover, after the recombination process, this strain is also able to digest the contaminant miniplasmid through endonucleases action. A highest mcDNAp53/ PP-p53 ratio was achieved by optimizing the recombination step to use 0.01% of Larabinose and 2h of induction. After the production, the mcDNA must be purified for further evaluation of transfection efficiency and cellular effect through in vitro studies. For that, a new chromatographic column was studied in order to explore the simultaneous affinity character of two ligands (arginine and lysine) immobilized in the same functional group (triazine), in which hydrophobic binding/elution conditions were explored and optimized in order to purify the sc mcDNA-p53. When it was used lower salt concentration in the equilibrium step, it seemed that the selectivity of the column was favored, obtaining a sample of sc mcDNA-p53 with lower content of impurities, although the total recovery of the injected sample has been sacrificed. On the other hand, using the strategy of effective binding of the sample to the column and manipulating the pH was seemed to favor the recovery of sc mcDNA-p53 despite containing some impurities. In vitro transfection studies were performed to evaluate the transfection efficiency of PP and mcDNA vectors and the p53 expression, by using HeLA cells. Through immunocytochemistry it was observed that PP and mcDNA vectors were able to transfect HeLa cells, being more efficient with mcDNA. By RT-PCR it was confirmed the presence of p53 transcripts and by western blot it was verified the correct expression of p53 tumor suppressor in transfected cells Overall, this work allowed the construction, biosynthesis and recovery of a minicircular DNA vector encoding the p53 tumor supressor protein, showing great potential to proceed to in vitro and in vivo studies in order to develop a strategy for cervical cancer treatment in the future.
- Sociodemographic Data and Work-Related Musculoskeletal Symptoms in the Metal Polishing Industry: A Case Study in Central PortugalPublication . Alves, Joel; Lima, Tânia M.; Gaspar, Pedro DinisThe prevalence of work-related musculoskeletal disorders is a red flag in industries and is considered an occupational health issue that affects the operator’s well-being, safety, and health. This study contributes to understanding the impact of Industry 5.0 on the workforce and emphasizes the importance of promoting a safe and healthy working environment. Thus, it focuses on the assessment of anthropometric characteristics and work-related musculoskeletal symptoms in a real industrial environment, specifically in the metal polishing sector of a metallurgical industry in Portugal. The research collected data on physical attributes, health status, and musculoskeletal symptoms using the Portuguese version of the Nordic Musculoskeletal Questionnaire, and incorporated sociodemographic data, including age, gender, and professional status, to provide a comprehensive understanding of the active workforce. The results were analyzed by gender and age clusters and revealed a prevalence of musculoskeletal symptoms, particularly in the spinal column and upper limbs, with a significant impact on the ability to perform work tasks. The need for preventive measures and ergonomic interventions to minimize the high predominance of musculoskeletal symptoms in Portuguese industries is also highlighted to improve the working conditions and enhance the well-being and health of operators.
- Indústria 5.0 – Priorização da Centralidade Humana para a Sustentabilidade e Resiliência na Produção IndustrialPublication . Alves, Joel; Lima, Tânia M.; Gaspar, Pedro DinisA Indústria 5.0 apresenta-se como uma estratégia que coloca o fator humano no centro da produção, priorizando o seu bem-estar, bem como a adoção de sistemas de produção mais sustentáveis e resilientes. Para a centralização humana, é necessário capacitar os seres humanos e, respetivamente, os operadores industriais, para melhorar as suas aptidões e competências individuais em colaboração ou cooperação com as tecnologias digitais. O principal objetivo e ponto distintivo desta investigação é determinar se a Indústria 5.0 é verdadeiramente orientada para o ser humano e como a centralidade humana pode ser criada com as tecnologias da Indústria 5.0. Para tal, este capítulo de revisão sistemática da literatura analisa e clarifica os conceitos e ideologia da Indústria 5.0, as suas respetivas tecnologias, assim como as estratégias de centralização humana, com o objetivo de alcançar sistemas sustentáveis e resilientes, especialmente para o trabalhador.
- A Chromatic Enzymatic Time-Temperature Integrator Device Based on the Degradation of Phenolic Compounds for the Real-Time Prediction of the Quality and Shelf Life of CherriesPublication . Gaspar, Pedro Dinis; Alves, Joel; Quelhas, Adriana Sebastião; Domingos, Christelle; Caio, Susana Matos SilvaThe particular characteristics of cherries, such as color, firmness, and palate increase their demand, as does, among other things, their antioxidant properties that benefit human health. However, their high perishability leads to a reduced shelf life and consequently generates undesirable changes in the cherry flow chain. To ensure food quality and safety and prevent food waste, a smart device prototype is proposed. The concepts related to the formulation and design of the enzymatic-type chromatic time-temperature integrator (TTI) device used to monitor the real-time quality of cherries are described. The kinetic parameters for thermal inactivation of cultivar Santina cherries were determined based on the degradation of phenolic compounds that are substrates of the polyphenol oxidase enzyme, whose hydroxylation reaction of a monophenol to o-diphenol leads to the oxidation in o-quinone. The proposed device concept aims to help retailers and consumers decide upon selling and buying according to the remaining shelf life, thus promoting sustainability related to food processes.
- A indústria 4.0 e o desafio sociodemográficoPublication . Alves, Joel Marques; Lima, Tânia Daniela Felgueiras de MirandaA era da digitalização impulsionada pela estratégia alemã, a Indústria 4.0, trouxe consigo a mudança de vários paradigmas, desde os problemas associados à sua implementação, aos novos modelos produtivos e à adaptação da força de trabalho a estas mudanças e alterações. Por outro lado, os problemas sociodemográficos estão cada vez mais presentes na atualidade, nomeadamente o envelhecimento da população e, consequentemente o prolongamento da vida ativa, tornando a força de trabalho existente nas empresas e indústrias mais envelhecida. Posto isto, uma vez que os sistemas produtivos são mais exigentes e a força de trabalho presente nas indústrias está cada vez mais envelhecida, neste trabalho são propostas medidas para colocar o fator humano no centro da produção, acompanhando, também, o desenvolvimento da Indústria 4.0. Mais especificamente, a adaptação e envolvimento dos operadores mais velhos nos novos sistemas produtivos digitais. Para isso, é descrito o ambiente das futuras indústrias, as smart factorires, e também é apresentada a comparação dos trabalhadores mais velhos com o conceito do Operador 4.0, de forma a que seja possível coexistir uma produtividade elevada com a força de trabalho existente nas unidades fabris.
- Fostering Awareness on Environmentally Sustainable Technological Solutions for the Post-Harvest Food Supply ChainPublication . Fernandez, Carlos M.; Alves, Joel; Gaspar, Pedro Dinis; Lima, Tânia M.This study presents a current status and future trends of innovative and environmentally sustainable technological solutions for the post-harvest food supply chain and the food industry, in terms of ecological packaging, active, and/or intelligent packaging. All these concerns are currently highlighted due to the strong increase in the purchase/sale of products on online platforms, as well as the requirements for stricter food security and safety. Thus, this study aims to increase the global awareness of agro-industrial micro, small, and medium size enterprises for the adoption of innovative food solutions though industry digitalization (Industry 4.0), associated logistics and circular economy, with a concern for cybersecurity and products information, communication and shelf-life extension. The adoption of these guidelines will certainly foster along the complete food chain (from producer to consumer, with all intermediary parties) the awareness on environmentally sustainable technological solutions for the post-harvest food supply chain, and thus, promoting the future food sustainability required by the population increase, the climate change, the exodus of rural population to urban areas, and food loss and waste.
- The sociodemographic challenge in human-centred production systems – a systematic literature reviewPublication . Alves, Joel; Lima, Tânia M.; Gaspar, Pedro Dinisindustries are currently struggling with ageing workforce in modern production systems associated with industry 4.0. The industrial socio-demographic problem is more and more present as the increas-ing of the ageing population results in the prolongation of the work-ing life and the consecutively in the ageing of the workforce in industries. This paper aims to conduct a systematic literature review on the challenges and concerns of ageing operators, including the physical, cognitive, ergonomic, and well-being conditions of the age-ing workforce in the industry 4.0 environment. The ScienceDirect, Scopus, web of Science and PubMed scientific databases were used to survey the studies and selected using PRiSMA guidelines. This paper was structured and analysed by clusters: Ageing, industry 4.0, Human Factors, and ergonomics. These clusters were developed as research lines: Ageing as the socio-demographic challenge, industry 4.0 as the technological development, Human Factors as the individual charac-teristics of the operator, and ergonomics as the work environment. Thus, human-centric approaches and ideas are discussed with the insights and issues of industry 4.0 technologies, Human Factors, and ergonomics to achieve a sustainable system at the engineering and social level.
- Innovative processes in smart packaging. A systematic reviewPublication . Fernandez, Carlos M.; Alves, Joel; Gaspar, Pedro Dinis; Lima, Tânia M.; Silva, Pedro Dinho daSmart packaging provides one possible solution that could reduce greenhouse gas emissions. In comparison with traditional packaging, which aims to extend the product's useful life and to facilitate transport and marketing, smart packaging allows increased efficiency, for example by ensuring authenticity and traceability from the product's origin, preventing fraud and theft, and improving security. Consequently, it may help to reduce pollution, food losses, and waste associated with the food supply chain. However, some questions must be answered to fully understand the advantages and limitations of its use. What are the most suitable smart packaging technologies for use in agro-industrial subsectors such as meat, dairy, fruits, and vegetables, bakery, and pastry? What are the opportunities from a perspective of life extension, process optimization, traceability, product quality, and safety? What are the future challenges? An up-to-date, systematic review was conducted of literature relevant to the application of indicator technologies, sensors, and data carriers in smart packaging, to answer these questions.
- Novel Design of Assistive Technologies Based on the Interconnection of Motion Capture and Virtual Reality Systems to Foster Task Performance of the Ageing WorkforcePublication . Alves, Joel; Lima, Tânia M.; Gaspar, Pedro DinisDemographic changes, increasing life expectancy, and increasing healthy life expectancy lead to an increasingly ageing workforce. This condition has a growing impact on the workforce in today’s industries. However, old workers are not a burden for companies and industries. They are a fundamental part of the industrial ecosystem due to the consistency of their human factors, namely their large knowledge, experience, and decision-making ability. For this reason, the ageing workforce must be incorporated and engaged through the introduction of digital age technologies to improve their working conditions and extend their active life. This paper focuses on a novel design of assistive technologies based on the interconnection of motion capture (MoCap) and virtual reality (VR) systems as a driver for the improvement of the task performance of the ageing workforce. We intend to explore how this technological tool can help and enhance the work of the ageing workforce. For this, we study two different areas of application: the training, learning, and communication of the older workers and the ergonomic analysis and workplace design for the ageing operators. In the end, a pilot study is proposed to apply this technology in real work environments.
- Simplified Approach to Predict Food Safety through the Maximum Specific Bacterial Growth Rate as Function of Extrinsic and Intrinsic ParametersPublication . Gaspar, Pedro Dinis; Alves, Joel; Pinto, Pedro Miguel MoreiraCurrently, we assist the emergence of sensors and low-cost information and communication technologies applied to food products, in order to improve food safety and quality along the food chain. Thus, it is relevant to implement predictive mathematical modeling tools in order to predict changes in the food quality and allow decision-making for expiration dates. To perform that, the Baranyi and Roberts model and the online tool Combined Database for Predictive Microbiology (Combase) were used to determine the factors that define the growth of different bacteria. These factors applied to the equation that determines the maximum specific growth rate establish a relation between the bacterial growth and the intrinsic and extrinsic factors that define the bacteria environment. These models may be programmed in low-cost wireless biochemical sensor devices applied to packaging and food supply chains to promote food safety and quality through real time traceability.