Leveraging the future of diagnosis and management of diabetes: From old indexes to new technologies.

BACKGROUND: Diabetes is a heterogeneous and multifactorial disease. However, glycemia and glycated hemoglobin have been the focus of diabetes diagnosis and management for the last decades. As diabetes management goes far beyond glucose control, it has become clear that assessment of other biochemical parameters gives a much wider view of the metabolic state of each individual, enabling a precision medicine approach. METHODS: In this review, we summarize and discuss indexes that have been used in epidemiological studies and in the clinical practice. RESULTS: Indexes of insulin secretion, sensitivity/resistance and metabolism have been developed and validated over the years to account also with insulin, C-peptide, triglycerides or even anthropometric measures. Nevertheless, each one has their own objective and consequently, advantages and disadvantages for specific cases. Thus, we discuss how new technologies, namely new sensors but also new softwares/applications, can improve the diagnosis and management of diabetes, both for healthcare professionals but also for caretakers and, importantly, to promote the empowerment of people living with diabetes. CONCLUSIONS: In long-term, the solution for a better diabetes management would be a platform that allows to integrate all sorts of relevant information for the person with diabetes and for the healthcare practitioners, namely glucose, insulin and C-peptide or, in case of need, other parameters/indexes at home, sometimes more than once a day. This solution would allow a better and simpler disease management, more adequate therapeutics thereby improving patients' quality of life and reducing associated costs.

Lack of mitochondrial NADP(H)-transhydrogenase expression in macrophages exacerbates atherosclerosis in hypercholesterolemic mice.

The atherosclerosis prone LDL receptor knockout mice (Ldlr-/-, C57BL/6J background) carry a deletion of the NADP(H)-transhydrogenase gene (Nnt) encoding the mitochondrial enzyme that catalyzes NADPH synthesis. Here we hypothesize that both increased NADPH consumption (due to increased steroidogenesis) and decreased NADPH generation (due to Nnt deficiency) in Ldlr-/- mice contribute to establish a macrophage oxidative stress and increase atherosclerosis development. Thus, we compared peritoneal macrophages and liver mitochondria from three C57BL/6J mice lines: Ldlr and Nnt double mutant, single Nnt mutant and wild-type. We found increased oxidants production in both mitochondria and macrophages according to a gradient: double mutant > single mutant > wild-type. We also observed a parallel up-regulation of mitochondrial biogenesis (PGC1a, TFAM and respiratory complexes levels) and inflammatory (iNOS, IL6 and IL1b) markers in single and double mutant macrophages. When exposed to modified LDL, the single and double mutant cells exhibited significant increases in lipid accumulation leading to foam cell formation, the hallmark of atherosclerosis. Nnt deficiency cells showed up-regulation of CD36 and down-regulation of ABCA1 transporters what may explain lipid accumulation in macrophages. Finally, Nnt wild-type bone marrow transplantation into LDLr-/- mice resulted in reduced diet-induced atherosclerosis. Therefore, Nnt plays a critical role in the maintenance of macrophage redox, inflammatory and cholesterol homeostasis, which is relevant for delaying the atherogenesis process.

Coenzyme Q10 protects against ß-cell toxicity induced by pravastatin treatment of hypercholesterolemia.

New onset of diabetes is associated with the use of statins. We have recently demonstrated that pravastatin-treated hypercholesterolemic LDL receptor knockout (LDLr-/- ) mice exhibit reductions in insulin secretion and increased islet cell death and oxidative stress. Here, we hypothesized that these diabetogenic effects of pravastatin could be counteracted by treatment with the antioxidant coenzyme Q 10 (CoQ 10 ), an intermediate generated in the cholesterol synthesis pathway. LDLr -/- mice were treated with pravastatin and/or CoQ 10 for 2 months. Pravastatin treatment resulted in a 75% decrease of liver CoQ 10 content. Dietary CoQ 10 supplementation of pravastatin-treated mice reversed fasting hyperglycemia, improved glucose tolerance (20%) and insulin sensitivity (>2-fold), and fully restored islet glucose-stimulated insulin secretion impaired by pravastatin (40%). Pravastatin had no effect on insulin secretion of wild-type mice. In vitro, insulin-secreting INS1E cells cotreated with CoQ 10 were protected from cell death and oxidative stress induced by pravastatin. Simvastatin and atorvastatin were more potent in inducing dose-dependent INS1E cell death (10-15-fold), which were also attenuated by CoQ 10 cotreatment. Together, these results demonstrate that statins impair ß-cell redox balance, function and viability. However, CoQ 10 supplementation can protect the statins detrimental effects on the endocrine pancreas.

Preparation of Porous Scaffold Based on Poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) and FucoPol.

This work focused on the development of porous scaffolds based on biocomposites comprising two biodegradable and biocompatible biopolymers: a terpolyester, poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-3-hydroxyhexanoate) (PHBHVHHx), and the bacterial polysaccharide FucoPol. The PHBHVHHx terpolymer was composed of 3-hydroxybutyrate (55 wt%), 3-hydroxyvalerate (21 wt%), and 3-hydroxyhexanoate (24 wt%). This hydrophobic polyester has low crystallinity and can form elastic and flexible films. Fucopol is a fucose-containing water-soluble polysaccharide that forms viscous solutions with shear thinning behavior and has demonstrated emulsion-forming and stabilizing capacity and wound healing ability. Emulsion-templating was used to fabricate PHA-based porous structures in which FucoPol acted as a bioemulsifier. Compared with the scaffolds obtained from emulsions with only water, the use of FucoPol aqueous solutions resulted in structures with improved mechanical properties, namely higher tensile strength (4.4 MPa) and a higher Young's Modulus (85 MPa), together with an elongation at break of 52%. These features, together with the scaffolds' high porosity and pore interconnectivity, suggest their potential to sustain cell adhesion and proliferation, which is further supported by FucoPol's demonstrated wound healing ability. Therefore, the developed PHBHVHHx:FucoPol scaffolds arise as innovative porous bioactive structures with great potential for use in tissue engineering applications.

SARS-CoV-2 virus-like-particles via liposomal reconstitution of spike glycoproteins.

The SARS-CoV-2 virus, implicated in the COVID-19 pandemic, recognizes and binds host cells using its spike glycoprotein through an angiotensin converting enzyme 2 (ACE-2) receptor-mediated pathway. Recent research suggests that spatial distributions of the spike protein may influence viral interactions with target cells and immune systems. The goal of this study has been to develop a liposome-based virus-like particle (VLP) by reconstituting the SARS-CoV-2 spike glycoprotein within a synthetic nanoparticle membrane, aiming to eventually establish tunability in spike protein presentation on the nanoparticle surface. Here we report on first steps to this goal, wherein liposomal SARS-CoV-2 VLPs were successfully produced via detergent mediated spike protein reconstitution. The resultant VLPs are shown to successfully co-localize in vitro with the ACE-2 receptor on lung epithelial cell surfaces, followed by internalization into these cells. These VLPs are the first step toward the overall goal of this research which is to form an understanding of the relationship between spike protein surface density and cell-level immune response, eventually toward creating better vaccines and anti-viral therapeutics.

Leucine-Rich Diet Improved Muscle Function in Cachectic Walker 256 Tumour-Bearing Wistar Rats.

Skeletal muscle atrophy occurs in several pathological conditions, such as cancer, especially during cancer-induced cachexia. This condition is associated with increased morbidity and poor treatment response, decreased quality of life, and increased mortality in cancer patients. A leucine-rich diet could be used as a coadjutant therapy to prevent muscle atrophy in patients suffering from cancer cachexia. Besides muscle atrophy, muscle function loss is even more important to patient quality of life. Therefore, this study aimed to investigate the potential beneficial effects of leucine supplementation on whole-body functional/movement properties, as well as some markers of muscle breakdown and inflammatory status. Adult Wistar rats were randomly distributed into four experimental groups. Two groups were fed with a control diet (18% protein): Control (C) and Walker 256 tumour-bearing (W), and two other groups were fed with a leucine-rich diet (18% protein + 3% leucine): Leucine Control (L) and Leucine Walker 256 tumour-bearing (LW). A functional analysis (walking, behaviour, and strength tests) was performed before and after tumour inoculation. Cachexia parameters such as body weight loss, muscle and fat mass, pro-inflammatory cytokine profile, and molecular and morphological aspects of skeletal muscle were also determined. As expected, Walker 256 tumour growth led to muscle function decline, cachexia manifestation symptoms, muscle fibre cross-section area reduction, and classical muscle protein degradation pathway activation, with upregulation of FoxO1, MuRF-1, and 20S proteins. On the other hand, despite having no effect on the walking test, inflammation status or muscle oxidative capacity, the leucine-rich diet improved muscle strength and behaviour performance, maintained body weight, fat and muscle mass and decreased some protein degradation markers in Walker 256 tumour-bearing rats. Indeed, a leucine-rich diet alone could not completely revert cachexia but could potentially diminish muscle protein degradation, leading to better muscle functional performance in cancer cachexia.

Translation, cross-cultural adaptation and psychometric evaluation of Brazilian Portuguese version of the 14-item Health Literacy Scale. / Tradução, adaptação transcultural e avaliação psicométrica da versão em português (brasileiro) do 14-item Health Literacy Scale.

We evaluated the psychometric properties of the Health Literacy Scale - 14 (HLS-14), the Brazilian Portuguese version. In the methodological study with a cross-sectional design, the following were performed: translation, cross-cultural adaptation and evaluation of the psychometric properties. After being subjected to a committee of experts, translated and adapted, the instrument was pre-tested with 52 adults and applied to 143 adults and older people of Piracicaba-SP. Internal consistency was assessed based on the Kendall correlation coefficient and Cronbach's alpha (>0.70), and the confirmatory factor analysis (CFA) was conducted using the chi-square test, the Standardized Residual Mean Square Root (SRMR), the Root Mean Square Error of Approximation (RMSEA) (<0.05), the Comparative Fit Index (CFI) (>0.95) and the Tucker-Lewis Index (TLI) (>0.95). The analysis of operational equivalence showed agreement between most of the questions, exception questions 6, 8, 9 and 14. Cronbach's alpha was 0.82. There was reasonable adjustment in the CFA: CFI=0.886, TLI=0.86, RMSEA=0.085 (90%CI: 0.065-0.105), SRMR=0.071, chi-square (74 degrees of freedom) = 149.510, p<0.001. The exclusion of question 5 increased the adjustment level to satisfactory. The Brazilian Portuguese version of HLS-14 can be considered a valid health literacy assessment instrument.

Foram avaliadas as propriedades psicométricas da versão brasileira do 14-item Health Literacy Scale (HLS-14). No estudo metodológico com delineamento transversal realizou-se tradução, adaptação transcultural e avaliação das propriedades psicométricas. Depois de passar por comitê de especialistas, traduzido e adaptado, o instrumento foi pré-testado em 52 adultos, e aplicado em 143 adultos e idosos de Piracicaba-SP. A consistência interna foi avaliada através do coeficiente de correlação de Kendall e α de Cronbach (>0,70) e a análise fatorial confirmatória (AFC) por meio do teste de qui-quadrado, raiz quadrada média residual padronizada (SRMR), raiz da média dos quadrados dos erros de aproximação (RMSEA) (<0,05), índice de ajuste comparativo (CFI) (>0,95) e índice de Tucker-Lewis (TLI) (>0,95). A equivalência operacional apresentou concordância entre as questões, com exceção das questões 6, 8, 9 e 14. O coeficiente α de Cronbach foi 0,82. Houve ajuste razoável na AFC, CFI=0,886, TLI=0,86, RMSEA=0,085 (IC90%: 0,065-0,105), SRMR=0,071, qui-quadrado (74 graus de liberdade) =149,510, p<0,001. A exclusão da questão 5 elevou os índices de ajuste a níveis satisfatórios. A versão brasileira do HLS-14 foi considerada válida para mensurar literacia em saúde.

Leucine-rich diet induces a shift in tumour metabolism from glycolytic towards oxidative phosphorylation, reducing glucose consumption and metastasis in Walker-256 tumour-bearing rats.

Leucine can stimulate protein synthesis in skeletal muscle, and recent studies have shown an increase in leucine-related mitochondrial biogenesis and oxidative phosphorylation capacity in muscle cells. However, leucine-related effects in tumour tissues are still poorly understood. Thus, we described the effects of leucine in both in vivo and in vitro models of a Walker-256 tumour. Tumour-bearing Wistar rats were randomly distributed into a control group (W; normoprotein diet) and leucine group (LW; leucine-rich diet [normoprotein + 3% leucine]). After 20 days of tumour evolution, the animals underwent 18-fludeoxyglucose positron emission computed tomography (18F-FDG PET-CT) imaging, and after euthanasia, fresh tumour biopsy samples were taken for oxygen consumption rate measurements (Oroboros Oxygraph), electron microscopy analysis and RNA and protein extraction. Our main results from the LW group showed no tumour size change, lower tumour glucose (18F-FDG) uptake, and reduced metastatic sites. Furthermore, leucine stimulated a shift in tumour metabolism from glycolytic towards oxidative phosphorylation, higher mRNA and protein expression of oxidative phosphorylation components, and enhanced mitochondrial density/area even though the leucine-treated tumour had a higher number of apoptotic nuclei with increased oxidative stress. In summary, a leucine-rich diet directed Walker-256 tumour metabolism to a less glycolytic phenotype profile in which these metabolic alterations were associated with a decrease in tumour aggressiveness and reduction in the number of metastatic sites in rats fed a diet supplemented with this branched-chain amino acid.

BigR is a sulfide sensor that regulates a sulfur transferase/dioxygenase required for aerobic respiration of plant bacteria under sulfide stress.

To cope with toxic levels of H2S, the plant pathogens Xylella fastidiosa and Agrobacterium tumefaciens employ the bigR operon to oxidize H2S into sulfite. The bigR operon is regulated by the transcriptional repressor BigR and it encodes a bifunctional sulfur transferase (ST) and sulfur dioxygenase (SDO) enzyme, Blh, required for H2S oxidation and bacterial growth under hypoxia. However, how Blh operates to enhance bacterial survival under hypoxia and how BigR is deactivated to derepress operon transcription is unknown. Here, we show that the ST and SDO activities of Blh are in vitro coupled and necessary to oxidize sulfide into sulfite, and that Blh is critical to maintain the oxygen flux during A. tumefaciens respiration when oxygen becomes limited to cells. We also show that H2S and polysulfides inactivate BigR leading to operon transcription. Moreover, we show that sulfite, which is produced by Blh in the ST and SDO reactions, is toxic to Citrus sinensis and that X. fastidiosa-infected plants accumulate sulfite and higher transcript levels of sulfite detoxification enzymes, suggesting that they are under sulfite stress. These results indicate that BigR acts as a sulfide sensor in the H2S oxidation mechanism that allows pathogens to colonize plant tissues where oxygen is a limiting factor.

Malacofauna bentônica do Lago Igapó, Londrina (Paraná, Brasil), com ênfase na espécie invasora mexilhão-dourado Limnoperna fortunei (Dunker, 1857) / Benthic malacofauna from the Igapó Lake, Londrina (Paraná, Brazil), with emphasis on the invasive species Golden Mussel Limnoperna fortunei (Dunker, 1857)

Objetivo: quantificar a abundância e a biomassa de moluscos bentônicos no Lago Igapó I, Londrina, Paraná, Brasil. Material e Métodos: foram realizadas duas coletas no Lago Igapó I, a primeira em junho de 2015 e a segunda em fevereiro de 2016. O substrato (incluindo os moluscos incrustados) foi amostrado utilizando um quadrante com área de 1 m2, onde 10 amostragens foram realizadas entre três pontos distintos do lago. Os moluscos capturados foram anestesiados e eutanasiados por superexposição ao gelo. Posteriormente, o material foi quantificado em abundância (n) e biomassa total (kg), e armazenado em tambores contendo formol 4% tamponado com carbonato de cálcio. Resultados: foram identificadas cinco espécies de moluscos, sendo três não nativas (Limnoperna fortunei, Corbicula fluminea e Melanoides tuberculata), uma nativa (Aylacostoma cf. tenuilabris) e um indivíduo do gênero Pomacea. Em ambas as coletas, L. fortunei compreendeu aproximadamente 90% da abundância e biomassa total. A partir da densidade média de L. fortunei e a área total do Lago Igapó I, estimou-se que a população total de mexilhões-dourados pode chegar a 633 milhões de indivíduos, correspondendo a 638 toneladas de biomassa. Conclusão: é evidente a dominância da espécie invasora L. fortunei no Lago Igapó I, onde esta pode causar diversos efeitos negativos, como alterações no ciclo de nutrientes, redução de espécies nativas, introdução de parasitos, bioacumulação de metais pesados na cadeia trófica, diminuição da qualidade da água para uso humano e obstrução de encanamentos com risco de alagamentos. Desta forma, recomenda-se uma imediata ação de manejo neste ambiente para retirada de indivíduos da espécie, com consequente redução de sua abundância.(AU)

Objective: quantify the abundance and biomass of benthic molluscs in Igapó Lake I, Londrina, Paraná, Brazil. Material and Methods: two samplings was made at Igapó Lake I, first in June 2015 and then in February 2016. The substrate (including all embedded molluscs) was sampled using a 1 m2 quadrant, where 10 samples were taken between three distinct points from the lake. All molluscs captured were anesthetized and euthanized by overexposure to ice. Subsequently, the material was quantified in quantity (n) and total biomass (kg), and stored in barrels containing 4% formaldehyde buffered with calcium carbonate. Results: five species of molluscs were identified, three non native species (Limnoperna fortunei, Corbicula fluminea and Melanoides tuberculata), one native (Aylacostoma cf. tenuilabris) and one Pomacea sp. In both sampling, L. fortunei comprised approximately 90% of the abundance and total biomass. From the average density of L. fortunei and the total area of the Igapó Lake I, it was estimated that the total population of golden mussels can reach 633 million individuals, corresponding to 638 tons of biomass. Conclusion: the dominance of the invasive species L. fortunei in Igapó Lake I is evident, where it can cause several negative effects, such as alterations in the nutrient cycle, reduction of native species, introduction of parasites, bioaccumulation of heavy metals in the food chain, decreased quality of water for human use and obstruction of pipes obstruction with risk of overflow. Therefore, we recommend an immediate management action in this environment in order to remove individuals of this species and, consequently, to reduce its abundance. (AU)

Relações familiares, álcool e outras drogas: uma revisão integrativa / Family relationships, alcohol and other drugs: integrative review

Trata-se de uma revisão integrativa objetivando responder a seguinte pergunta: "Qual o conhecimento científico produzido sobre as relações familiares e problemática do uso, abuso e dependência de álcool e outras drogas, principalmente quanto ao relacionado a fatores de risco e proteção?" Objetivo da pesquisa: analisar a produção científica sobre as relações familiares e uso, abuso e dependência de álcool e outras drogas. Método: revisão de literatura nas bases de dados BIREME (SciELO, LILACS, PubMed (MEDLINE) e Psycinfo. Resultados: Foram encontrados 2.625 artigos, sendo selecionados 15 artigos, (10) PubMed (MEDLINE) e (5) Scielo. Houve predomínio de 12 publicações a partir de 2009. Conclusão: a comunicação familiar pode ser considerada tanto situação de risco como de proteção, requerendo dos profissionais de saúde uma escuta qualificada e habilidades para intervenção terapêutica junto à família nesse tema.

This is an integrative review aimed to answer the following question: "What is the scientific knowledge produced on family relationships and problematic use, abuse and dependence on alcohol and other drugs, especially in their risk and protective factors?" The purpose of the research was to analyze the scientific literature on family relationships and use, abuse and dependence on alcohol and other drugs. Method: literature review in BIREME databases (SciELO, LILACS, PubMed (MEDLINE) and PsycINFO. Results: There were 2,625 articles found, and 15 articleswere selected: (10) PubMed (MEDLINE ) and (5) Scielo. There were 12 publications predominating from 2009. Conclusion: family communication can be considered both risk and protection by requiring health professionals qualified listening and skills for therapeutic intervention with the family in this area.