Curcumina e um novo complexo Ruteno(II)curcumina : caracterização e avaliação das suas potencialidades biológicas

Mestrado em Bioquímica - Métodos BiomolecularesA curcumina é um pigmento de cor amarela, da família dos polifenóis, obtido a partir dos rizomas da planta Curcuma longa. É dotada de diversas propriedades biológicas que se relacionam com a vasta gama de alvos moleculares que possui, sendo de destacar a sua atividade anticancerígena já provada em diversas linhas celulares cancerígenas. No entanto, a atividade biológica da curcumina é limitada pela sua baixa biodisponibilidade que se deve principalmente à sua fraca solubilidade e rápida degradação em condições fisiológicas. Uma estratégia para contornar as limitações terapêuticas da curcumina é a sua complexação com iões metálicos, nomeadamente com complexos de ruténio(II). Neste trabalho foi preparado um novo complexo de curcumina com ruténio(II) e tritiaciclononano [9anoS3] com o objetivo de aumentar a solubilidade da curcumina em meio fisiológico. A curcumina foi extraída e purificada a partir de rizomas secos e pulverizados de Curcuma longa sendo depois desprotonada e coordenada com o precursor [Ru(II)(9anoS3)(DMSO)Cl2] (DMSO=dimetilsulfóxido). A estrutura e pureza do complexo formado, [Ru(II)(9aneS3)(curcumina)(S-DMSO)]Cl foi avaliada por Ressonância Magnética Nuclear de 1H e 13C, espectrometria de massa e análise elementar. A afinidade do novo complexo para ligação ao ácido desoxirribonucleico (ADN) foi estudada por ensaios de titulação e por determinação da variação na temperatura de desnaturação do ADN de esperma de salmão. Este estudo permitiu determinar que o complexo tem a capacidade de intercalar com o ADN com uma constante de ligação comparável aos intercaladores de ADN já conhecidos (4.00x105 M-1). A atividade citotóxica deste complexo contra o cancro de próstata foi estudada in vitro, usando como controlo positivo a curcumina. Mais ainda, aproveitando o potencial fotossensibilizador da curcumina, os ensaios foram feitos na presença e na ausência de luz. Para isso, usaram-se culturas celulares humanas isoladas de carcinoma de próstata (PC-3) e a toxicidade foi avaliada em linhas celulares prostáticas não tumorais (PNT-2). Com estes ensaios verificou-se que o complexo preparado não tem efeito citotóxico nem fototóxico nas concentrações usadas para as linhas celulares estudas. Mais, observou-se que a curcumina apresenta, no escuro, efeito citotóxico nas concentrações usadas e ainda, que este efeito é fortemente potenciado na presença de luz, sendo um potencial agente para terapia fotodinâmica em cancro de próstata.Curcumin is a yellow pigment, of the family of polyphenols, obtained from the rhizomes of the Curcuma longa. It is provided with several biological properties which relate with the wide range of molecular targets that possesses, especially the anticancer activity already proven in various cell lines. However, the biological activity of curcumin is limited by their low bioavailability which is mainly because their poor solubility and rapid degradation in physiological conditions. One promising strategy to circumvent the therapeutic limitations of curcumin is the binding with metal ions, namely with ruthenium(II) complexes. In this work, was prepared a new complex of curcumin with ruthenium(II) and trithiacyclononane (9aneS3) with the aim of increasing the solubility of curcumin in physiological medium. The curcumina was extracted and purified from the powder rhizomes of Curcuma longa being after deprotonated and coordinated with the precursor [Ru(II)(9aneS3)(DMSO)Cl2] (DMSO=dimethylsulfoxide). The structure and purity of the prepared complex, [Ru(II)(9aneS3)(curcumina)(S-DMSO)]Cl was evaluated by 1H and 13C Nuclear Magnetic Resonance spectroscopies, mass spectrometry and elemental analysis. The affinity of the new complex to bind to deoxyribonucleic acid (DNA) was studied by titration assays and determination of the variation on sperm salmon DNA melting temperature. This study allowed to determine that the complex has ability to intercalate with DNA with a binding constant comparable with classical intercalators. The cytotoxic activity of this complex against prostate cancer was studied in vitro, using curcumin as a positive control. Moreover, taking advantage of the photosensitizer potential of curcumin, the assays were made under dark and light conditions. For this, human cell cultures were used isolated from prostate carcinoma (PC-3) and the toxicity was evaluated in non-tumour prostate cells (PNT-2). With these tests, it was found that the new complex do not have any cytotoxic or phototoxic effect in the same concentrations range tested for cell lines studied. Further, was observed that the curcumin presents, in dark, cytotoxic effect and, that this effect is strongly potentiated by light presence, with a potential for photodynamic therapy in prostate cancer

The Role of Endocrine-Disrupting Chemicals in Male Fertility Decline

Endocrine-disrupting chemicals (EDCs) are exogenous compounds with natural or anthropogenic origin omnipresent in the environment. These compounds disrupt endocrine function through interaction with hormone receptor or alteration of hormone synthesis. Humans are environmentally exposed to EDCs through the air, water, food and occupation. During the last decades, there has been a concern that exposure to EDCs may contribute to an impairment of human reproductive function. EDCs affect male fertility at multiple levels, from sperm production and quality to the morphology and histology of the male reproductive system. It has been proposed that exposure to EDCs may contribute to an impairment of sperm motility, concentration, volume and morphology and an increase in the sperm DNA damage. Moreover, EDCs exert reproductive toxicity inducing structural damage on the testis vasculature and blood-testis barrier and cytotoxicity on Sertoli and Leydig cells. This chapter will explore the effects of EDCs in male reproductive system and in the decline of male fertility

How to motivate students to learn Metabolic Biochemistry in a Biomedical Sciences curricula

[EN] Teaching methodologies used in biochemistry classes at the University level are traditionally dependent on theorical classes. The assessment is usually based on written tests performed at the end of the semester. However, most students who learn metabolism by this traditional method consider the study of metabolic biochemistry a terrifying and unforgettable experience. Understanding biochemical metabolic pathways was the proposed goal of the Medical Biochemistry curricular unit. To this end, the multi-method active learning approach was used in order to increase students’ motivation towards the learning process and to allow the development of skills associated with group conflict resolution, critical thinking and communication skills. Overall, students and learning facilitators were highly motivated by the diversity of learning activities, particularly due to the emphasis on correlating theoretical knowledge with human health and disease. As a quality control exercise, the students were asked to answer a questionnaire on their evaluation of the teaching/learning experience. Thus, the initial analysis of the student’s perception questionnaires permits to conclude that the approach undertaken yields results that surpass the traditional teaching methods. Investing in preparing attractive and motivating classes increases students and teacher’s general satisfaction and the learning/teaching process becomes more efficient.This work was also supported by an individual grant from FCT of the Portuguese Ministry of Science and Higher Education to M.C.H (SFRH/BD/131846/2017).Carvalho Henriques, M.; Fardilha, M. (2019). How to motivate students to learn Metabolic Biochemistry in a Biomedical Sciences curricula. En HEAD'19. 5th International Conference on Higher Education Advances. Editorial Universitat Politècnica de València. 1249-1254. https://doi.org/10.4995/HEAD19.2019.9315OCS1249125

ANALYSIS OF VANILLIN BY TLC AND HPLC-PDA IN HERBAL MATERIAL AND TINCTURE FROM Vanilla planifolia Jacks ex. Andrews

Vanilla planifolia (Orchidiaceae) is a species that is renowned globally and represents the largest source of vanillin flavoring used in the food, cosmetic and pharmaceutical industries. This study was carried out to analyze by TLC and HPLC-PDA vanillin in herbal drug and tincture from V. planifolia. The herbal drug was obtained with hydroalcoholic solution under reflux; and a kinetic reaction was performed by TLC. The influences of solvent and herbal drug concentration were studied through an experimental design. The solutions (herbal drug, tincture and standard – vanillin) were prepared and analyzed in HPLC coupled with DAD detector, using wavelength of 280 nm. The total extraction of vanillin was achieved after three extraction cycles, using 1.0 g of herbal material and Ethanol 50% (v/v) as solvent. The method was linear (R2> 0.99) and demonstrated repeatability (RSD < 0.90), intermediate precision (RSD < 1.09), recovery (93.12-113.74%), as well as robustness (RSD < 4.33). The total content of vanillin found was 1.82 g% and 0.21 g% for herbal drug and tincture, respectively. A simple and optimized method for sample preparation by reflux was able to provide the exhaustive extraction of vanillin and does not compromise the reliability of the HPLC-PDA method. The chromatographic procedure was validated to separate and quantify vanillin in herbal material and tincture from pods of V. planifolia

A ruthenium(II)-trithiacyclononane curcuminate complex: synthesis, characterization, DNA-interaction, and cytotoxic activity

The coordination of ruthenium(II) complexes to anionic oxygen-based donors are very rare. This study describes a simple, one-pot method for obtaining [ruthenium(II)(trithiacyclononane)(curcumin)(S-DMSO)]Cl (1) in 37% yield. The structural characterization of complex 1 by elemental analysis, FT-IR, 1-D and 2-D NMR, ESI+-MS as well as UV–vis and fluorescence spectroscopies are presented. The DNA-melting temperature (Tm) assay shows that salmon sperm DNA (smDNA) in the presence of complex 1 has a higher melting temperature, with ΔTm = 7.4 °C, while in the presence of curcumin the melting temperature remains unaltered. The in vitro cytotoxic activities of curcumin and complex 1 were investigated using the tumor human prostate cell line, PC-3, and the healthy cell line, PNT-2. Complex 1 is innocuous toward normal prostate epithelial cells and, whereas curcumin is toxic, with inhibition rates of ca. 35 and 65% at 50 and 80 μM, respectively. On the tumor cell line PC-3, complex 1 did not cause viability changes, whereas curcumin exhibited dose-dependent inhibition, with ca. 73% inhibition at the highest concentration tested, i.e. 80 μM. This study suggests that coordination with the trithiacyclononane ruthenium(II) scaffold stabilizes the photochemical properties of curcumin and strongly changes its biologic activity.publishe

Chagas disease and SARS-CoV-2 coinfection does not lead to worse in-hospital outcomes

Epidemiología; Microbiología; SARS-CoV-2Epidemiologia; Microbiologia; SARS-CoV-2Epidemiology; Microbiology; SARS-CoV-2Chagas disease (CD) continues to be a major public health burden in Latina America. Information on the interplay between COVID-19 and CD is lacking. Our aim was to assess clinical characteristics and in-hospital outcomes of patients with CD and COVID-19, and to compare it to non-CD patients. Consecutive patients with confirmed COVID-19 were included from March to September 2020. Genetic matching for sex, age, hypertension, diabetes mellitus and hospital was performed in a 4:1 ratio. Of the 7018 patients who had confirmed COVID-19, 31 patients with CD and 124 matched controls were included (median age 72 (64–80) years-old, 44.5% were male). At baseline, heart failure (25.8% vs. 9.7%) and atrial fibrillation (29.0% vs. 5.6%) were more frequent in CD patients than in the controls (p < 0.05). C-reactive protein levels were lower in CD patients compared with the controls (55.5 [35.7, 85.0] vs. 94.3 [50.7, 167.5] mg/dL). In-hospital management, outcomes and complications were similar between the groups. In this large Brazilian COVID-19 Registry, CD patients had a higher prevalence of atrial fibrillation and chronic heart failure compared with non-CD controls, with no differences in-hospital outcomes. The lower C-reactive protein levels in CD patients require further investigation.This study was supported in part by Minas Gerais State Agency for Research and Development (Fundação de Amparo à Pesquisa do Estado de Minas Gerais—FAPEMIG) [Grant Number APQ-00208-20], National Institute of Science and Technology for Health Technology Assessment (Instituto de Avaliação de Tecnologias em Saúde—IATS)/National Council for Scientific and Technological Development (Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq) [Grant Number 465518/2014-1], and CAPES Foundation (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) [Grant Number 88887.507149/2020-00]

TEORIA DA ADAPTAÇÃO E SAÚDE DO TRABALHADOR EM HOME OFFICE NA PANDEMIA DE COVID-19

Objetivo: refletir sobre as implicações do trabalho em Home Office no período da pandemia de COVID-19 na saúde dos indivíduos, sob a perspectiva da Teoria da Adaptação desenvolvida por Callista Roy. Método: estudo reflexivo baseado na aplicação da Teoria da Adaptação desenvolvida por Callista Roy relacionada às modificações do processo de trabalho impostas pela crise sanitária da pandemia de COVID-19, com ênfase no Home Office. Resultados: a Teoria da Adaptação de Callista Roy possui quatro modos adaptativos: fisiológico, autoconceito, desempenho de papel e interdependência. É possível verificar a interlocução de todas essas dimensões no trabalho em Home Office imposto pelo contexto da pandemia. Conclusão: a Teoria de Callista Roy subsidia as discussões sobre a possibilidade de adaptação neste novo contexto, seja de maneira pontual ou mediante transformações no processo de trabalho em longo prazo, superando limitações do indivíduo e descobrindo maneiras de se fazer e ser no campo do trabalho.Descritores: Saúde do Trabalhador. Teoria de Enfermagem. Enfermagem. Pandemias. Infecções por Coronavírus

Establishment and characterization of human pluripotent stem cells-derived brain organoids to model cerebellar diseases

Supplementary Information Te online version contains supplementary material available at https://doi.org/ 10.1038/s41598-022-16369-y.The establishment of robust human brain organoids to model cerebellar diseases is essential to study new therapeutic strategies for cerebellum-associated disorders. Machado-Joseph disease (MJD) is a cerebellar hereditary neurodegenerative disease, without therapeutic options able to prevent the disease progression. In the present work, control and MJD induced-pluripotent stem cells were used to establish human brain organoids. These organoids were characterized regarding brain development, cell type composition, and MJD-associated neuropathology markers, to evaluate their value for cerebellar diseases modeling. Our data indicate that the organoids recapitulated, to some extent, aspects of brain development, such as astroglia emerging after neurons and the presence of ventricular-like zones surrounded by glia and neurons that are found only in primate brains. Moreover, the brain organoids presented markers of neural progenitors proliferation, neuronal differentiation, inhibitory and excitatory synapses, and firing neurons. The established brain organoids also exhibited markers of cerebellar neurons progenitors and mature cerebellar neurons. Finally, MJD brain organoids showed higher ventricular-like zone numbers, an indication of lower maturation, and an increased number of ataxin-3-positive aggregates, compared with control organoids. Altogether, our data indicate that the established organoids recapitulate important characteristics of human brain development and exhibit cerebellar features, constituting a resourceful tool for testing therapeutic approaches for cerebellar diseases.This work was funded by the European Regional Development Fund (ERDF) through the Centro 2020 Regional Operational Programme under BrainHealth2020 projects (CENTRO-01-0145-FEDER-000008), through the COMPETE 2020—Operational Programme for Competitiveness and Internationalization and Portuguese national funds via FCT—Fundação para a Ciência e a Tecnologia, under projects — UIDB/04539/2020 and UIDP/04539/2020, POCI-01-0145-FEDER-030737 (NeuroStemForMJD, PTDC/BTM-ORG/30737/2017), CEEC-IND/04242/2017, PhD Scholarships 2020.04751.BD and 2020.07385.BD. It was also funded by the National Ataxia Foundation, the French Muscular Dystrophy Association (AFM-Téléthon) Trampoline Grant #20126, EU Joint Programme—Neurodegenerative Disease Research (JPND) Project JPCO FUND/0005/2015-ModelPolyQ), and the Richard Chin and Lily Lock Machado-Joseph Disease Research Fund.info:eu-repo/semantics/publishedVersio

Programa nacional de imunização:: a política de introdução de novas vacinas

O&nbsp;Objetivo&nbsp;deste trabalho foi&nbsp;relatar a implementação do Programa Nacional de Imunizações do SUS e os critérios para a incorporação de novas vacinas.&nbsp;Metodologia:&nbsp;relato de experiência descritivo sobre a implementação do PNI, o histórico de incorporações de novas vacinas e o Calendário Nacional de Vacinação.&nbsp;Resultados:&nbsp;O país apresenta um histórico de avanços na imunização da população brasileira, oferecendo o maior número de vacinas gratuitamente. São considerados critérios epidemiológicos, de eficácia, segurança e sócio-econômicos para a incorporação das vacinas, analisados pelo Comitê Técnico Assessor de Imunizações para posterior submissão à análise da CONITEC. A implementação depende do fornecimento sustentável amparado no fortalecimento do complexo industrial, visando a garantia da produção nacional de insumos com o envolvimento de laboratórios públicos. Os mecanismos adotados são o estímulo da produção nacional e as Parcerias para o Desenvolvimento Produtivo.&nbsp;Conclusões:&nbsp;As campanhas de vacinação têm alcance nacional, com calendários específicos instituídos. O programa vem empenhando esforços para atingir as metas com qualidade e segurança, consolidando o parque industrial para estabilidade de oferta das vacinas. A política de introdução de novas vacinas permite sua incorporação ao Calendário Nacional de forma sustentável, adequada às necessidades do país, à luz das evidências científicas

Assets acquisition planning

At the 127th European Study Group with Industry an energy sector com- pany proposed an industrial challenge that consisted on the asset acquisi- tion planning for its liquefied petroleum gas (LPG) cylinder business, one of the most recent business areas in the company. This business area is still in a growing phase and to satisfy the market needs and assure a sustainable growth a very tight control of the main assets, the LPG cylinders, is of paramount importance. Therefore, a detailed planning of all the as- sets acquisition is required, taking into consideration several variables: sales growth rate, seasonality, cylinder rotation and corresponding return rate to the filling plant. The challenge was to develop a model for the assets acquisition planning. In order to tackle this challenge, it was necessary to forecast the demand. For that purpose, time series techniques were used, in particular, moving averages and exponential smoothing. The results show that the seasonality does not explain all the variation of the demand, therefore it is necessary to use a model that would consider other possible explanatory variables. According to several authors, gas consumption may be influenced by several aspects, such as, atmospheric temperatures, heliophany (a measure of the day luminosity), wind, relative humidity, rains, minimum and maximum temperatures, demand in previous periods, and prices. The forecast of bottled propane gas sales and return rate was also addressed through multivariate linear regression. Regression models for the monthly number of bottles of types A and B were obtained, having pre- sented good percentages of explained variability with the variables under study. The main goal of the challenge, the acquisition plan, was addressed u ing inventory models with reverse logistics. Several deterministic approaches have been considered to enable different aspects in the framework. A new inventory model has been developed to contemplate the three possible des- tinations of returned bottles: cleaning, requalification, or disposal. The models were implemented in Excel and can be tested, using PRIO estimates of holding costs and fixed setup costs, and the forecasts of sales and return rate computed previously