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SARS-CoV-2 can induce vascular dysfunction and thrombotic events in patients with severe COVID-19; however, the cellular and molecular mechanisms behind these effects remain largely unknown. In this study, we used a combination of experimental and in silico approaches to investigate the role of PC in vascular and thrombotic events in COVID-19. Single-cell RNA-sequencing data from patients with COVID-19 and healthy subjects were obtained from the publicly available Gene Expression Omnibus (GEO) repository. In addition, HUVECs were treated with inactive protein C before exposure to SARS-CoV-2 infection or a severe COVID-19 serum. An RT-qPCR array containing 84 related genes was used, and the candidate genes obtained were evaluated. Activated protein C levels were measured using an ELISA kit. We identified at the single-cell level the expression of several pro-inflammatory and pro-coagulation genes in endothelial cells from the patients with COVID-19. Furthermore, we demonstrated that exposure to SARS-CoV-2 promoted transcriptional changes in HUVECs that were partly reversed by the activated protein C pretreatment. We also observed that the serum of severe COVID-19 had a significant amount of activated protein C that could protect endothelial cells from serum-induced activation. In conclusion, activated protein C protects endothelial cells from pro-inflammatory and pro-coagulant effects during exposure to the SARS-CoV-2 virus.

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The development of diabetic foot ulcers is a common and severe complication of diabetes that can significantly affect quality of life. The physiological healing cascade does not progress tissue repair in diabetic foot ulcerations in a timely manner. Serum markers from foot ulcers have been used to characterize the healing process of the diabetic foot using various collection techniques. This study aimed to compare the use of cervical brushes and the Levine technique to collect wound fluid from foot ulcers of people with diabetes in order to determine the presence of cytokines. The collected material was used for gene expression analysis of macrophage/monocyte-associated cytokines IL1-ß, IL-6, TNF-α, regulatory cytokine IL-10 and growth factor TGFß, via quantitative polymerase chain reaction (qPCR). Both collection methods produced sufficient amounts of RNA, but significantly more RNA was collected using a cervical brush (brush 224.82 ng/µL vs. Levine 80.90 ng/µL p = 0.0001). Significantly higher levels of expression of the following cytokine genes were detected in samples collected using a cervical brush: IL1-ß (p = 0.0001), IL-6 (p = 0.0106), IL-10 (p = 0.0277) and TGFß (p = 0.0002). Understanding why some wounds are difficult to heal is important for developing more effective treatments, and biomarkers may be useful for predicting the healing trajectory. These results demonstrate that it is possible to collect material from the wound bed for RT-qPCR analysis, and the cervical brush proved to be a simple and rapid method for monitoring cytokine gene expression.

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In the original publication, the authors claimed that Figure 6 reporting Western blot data was erroneous as published [...].

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Treatment of chronic wounds is challenging, and the development of different formulations based on insulin has shown efficacy due to their ability to regulate oxidative stress and inflammatory reactions. The formulation of insulin with polysaccharides in biohybrid hydrogel systems has the advantage of synergistically combining the bioactivity of the protein with the biocompatibility and hydrogel properties of polysaccharides. In this study, a hydrogel formulation containing insulin, chitosan, and hydroxypropyl methyl cellulose (Chi/HPMC/Ins) was prepared and characterized by FTIR, thermogravimetric, and gel point analyses. The in vitro cell viability and cell migration potential of the Chi/HPMC/Ins hydrogel were evaluated in human keratinocyte cells (HaCat) by MTT and wound scratch assay. The hydrogel was applied to excisional full-thickness wounds in diabetic mice for twenty days for in vivo studies. Cell viability studies indicated no cytotoxicity of the Chi/HPMC/Ins hydrogel. Moreover, the Chi/HPMC/Ins hydrogel promoted faster gap closure in the scratch assay. In vivo, the wounds treated with the Chi/HPMC/Ins hydrogel resulted in faster wound closure, formation of a more organized granulation tissue, and hair follicle regeneration. These results suggest that Chi/HPMC/Ins hydrogels might promote wound healing in vitro and in vivo and could be a new potential dressing for wound healing.

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The consumption of large amounts of dietary fats and pregnancy are independent factors that can promote changes in gut permeability and the gut microbiome landscape. However, there is limited evidence regarding the impact of pregnancy on the regulation of such parameters in females fed a high-fat diet. Here, gut permeability and microbiome landscape were evaluated in a mouse model of diet-induced obesity in pregnancy. The results show that pregnancy protected against the harmful effects of the consumption of a high-fat diet as a disruptor of gut permeability; thus, there was a two-fold reduction in FITC-dextran passage to the bloodstream compared to non-pregnant mice fed a high-fat diet (p < 0.01). This was accompanied by an increased expression of gut barrier-related transcripts, particularly in the ileum. In addition, the beneficial effect of pregnancy on female mice fed the high-fat diet was accompanied by a reduced presence of bacteria belonging to the genus Clostridia, and by increased Lactobacillus murinus in the gut (p < 0.05). Thus, this study advances the understanding of how pregnancy can act during a short window of time, protecting against the harmful effects of the consumption of a high-fat diet by promoting an increased expression of transcripts encoding proteins involved in the regulation of gut permeability, particularly in the ileum, and promoting changes in the gut microbiome.

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BACKGROUND: Photobiomodulation consists of inducing healing by irradiating light. This scoping review investigates the effect of blue light on the healing process. METHODS: The MEDLINE, Web of Science, Scopus, and CINAHL databases were searched. Two reviewers independently examined the search results and extracted data from the included studies. A descriptive analysis was performed. RESULTS: Twenty-two articles were included. Studies were categorized as in vitro/mixed, preclinical, and clinical. The power density used was 10-680 mW/cm2 in most of the in vitro/preclinical studies, the irradiation time ranged from 5 s to 10 min, and different wavelengths and energy densities were used. In clinical studies, the wavelength ranged from 405 to 470 nm, and the energy density varied from 1.5 to 30 J/cm2. CONCLUSIONS: A low energy density (<20 J/cm2) was able to stimulate the different cell types and proteins involved in healing, while a high energy density, 20.6-50 J/cm2, significantly reduced cell proliferation, migration, and metabolism. There is a great variety of device parameters among studies, and this makes it difficult to conclude what the best technical specifications are. Thus, further studies should be performed in order to define the appropriate parameters of light to be used.

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Obesity is one of the leading noncommunicable diseases in the world. Despite intense efforts to develop strategies to prevent and treat obesity, its prevalence continues to rise worldwide. A recent study has shown that the tricarboxylic acid intermediate succinate increases body energy expenditure by promoting brown adipose tissue thermogenesis through the activation of uncoupling protein-1; this has generated interest surrounding its potential usefulness as an approach to treat obesity. It is currently unknown how succinate impacts brown adipose tissue protein expression, and how exogenous succinate impacts body mass reduction promoted by a drug approved to treat human obesity, the glucagon-like-1 receptor agonist, liraglutide. In the first part of this study, we used bottom-up shotgun proteomics to determine the acute impact of exogenous succinate on the brown adipose tissue. We show that succinate rapidly affects the expression of 177 brown adipose tissue proteins, which are mostly associated with mitochondrial structure and function. In the second part of this study, we performed a short-term preclinical pharmacological intervention, treating diet-induced obese mice with a combination of exogenous succinate and liraglutide. We show that the combination was more efficient than liraglutide alone in promoting body mass reduction, food energy efficiency reduction, food intake reduction, and an increase in body temperature. Using serum metabolomics analysis, we showed that succinate, but not liraglutide, promoted a significant increase in the blood levels of several medium and long-chain fatty acids. In conclusion, exogenous succinate promotes rapid changes in brown adipose tissue mitochondrial proteins, and when used in association with liraglutide, increases body mass reduction.NEW & NOTEWORTHY Exogenous succinate induces major changes in brown adipose tissue protein expression affecting particularly mitochondrial respiration and structural proteins. When given exogenously in drinking water, succinate mitigates body mass gain in a rodent model of diet-induced obesity; in addition, when given in association with the glucagon-like peptide-1 receptor agonist, liraglutide, succinate increases body mass reduction promoted by liraglutide alone.

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Maternal obesity is an important risk factor for obesity, cardiovascular, and metabolic diseases in the offspring. Studies have shown that it leads to hypothalamic inflammation in the progeny, affecting the function of neurons regulating food intake and energy expenditure. In adult mice fed a high-fat diet, one of the hypothalamic abnormalities that contribute to the development of obesity is the damage of the blood-brain barrier (BBB) at the median eminence-arcuate nucleus (ME-ARC) interface; however, how the hypothalamic BBB is affected in the offspring of obese mothers requires further investigation. Here, we used confocal and transmission electron microscopy, transcript expression analysis, glucose tolerance testing, and a cross-fostering intervention to determine the impact of maternal obesity and breastfeeding on BBB integrity at the ME-ARC interface. The offspring of obese mothers were born smaller; conversely, at weaning, they presented larger body mass and glucose intolerance. In addition, maternal obesity-induced structural and functional damage of the offspring's ME-ARC BBB. By a cross-fostering intervention, some of the defects in barrier integrity and metabolism seen during development in an obesogenic diet were recovered. The offspring of obese dams breastfed by lean dams presented a reduction of body mass and glucose intolerance as compared to the offspring continuously exposed to an obesogenic environment during intrauterine and perinatal life; this was accompanied by partial recovery of the anatomical structure of the ME-ARC interface, and by the normalization of transcript expression of genes coding for hypothalamic neurotransmitters involved in energy balance and BBB integrity. Thus, maternal obesity promotes structural and functional damage of the hypothalamic BBB, which is, in part, reverted by lactation by lean mothers.NEW & NOTEWORTHY Maternal dietary habits directly influence offspring health. In this study, we aimed at determining the impact of maternal obesity on BBB integrity. We show that DIO offspring presented a leakier ME-BBB, accompanied by changes in the expression of transcripts encoding for endothelial and tanycytic proteins, as well as of hypothalamic neuropeptides. Breastfeeding in lean dams was sufficient to protect the offspring from ME-BBB disruption, providing a preventive strategy of nutritional intervention during early life.

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BACKGROUND: Impaired wound healing is a health problem around the world, and the search for a novel product to repair wounded skin is a major topic in the field. GW9508 is a synthetic molecule described as a selective agonist of free fatty acid receptors (FFARs) 1 and 4, and there is evidence of its anti-inflammatory effects on several organs of the body. PURPOSE: Here, we aimed to evaluate the effects of topical GW9508 on wound healing in mice. RESEARCH DESIGN: First, we used bioinformatic methods to determine the expression of FFAR1 and FFAR4 mRNA in the skin from a human cell atlas assembled with single-cell transcriptomes. Next, we employed 6-week-old C57BL6J mice with 2 wounds inflicted in the back. The mice were randomly divided into 2 groups, a control group, which received topical vehicle, and a treatment group, which received GW9508, for 12 days. The wound was monitored by photographic documentation every 2 days, and samples were collected at day 6 and 12 post injury for RT-PCR, western blot and histology analyses. RESULTS: FFAR1 and FFAR4 mRNA are expressed in skin cells in similar amounts to those in other tissues. Topical GW9508 accelerated wound healing and decreased gene expression of IL-10 and metalloproteinase 9 on days 6 and 12 post injury. It increased the quantity of Collagen I and improved the organization of collagen fibres. Conclusions: Our results show that GW9508 could be an attractive drug treatment for wounded skin. Future studies need to be performed to assess the impact of GW9508 in chronic wound models.

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O aumento significativo da longevidade na realidade brasileira teve como uma de suas consequências, o crescimento no número de pessoas dependentes que necessitam de cuidados de longa duração. Estes cuidados são prestados geralmente pelos familiares e devido às mudanças na estrutura e composição familiar e ao despreparo para a prestação desse cuidado, acabam acarretando sobrecarga física, financeira e emocional a esses cuidadores. O Centro Dia do idoso é um equipamento que se caracteriza como espaço de acolhimento, proteção e convivência de idosos dependentes cujas famílias não têm condições de prover atendimento durante todo o dia ou parte dele, oferecendo ainda apoio e orientação aos familiares sobre o cuidado com seu idoso. Até o momento são poucos os estudos sobre a percepção do bem estar subjetivo dos usuários desses serviços ofertados pela rede pública municipal. Objetivo: Analisar a contribuição dos cuidados oferecidos pelo Centro Dia para o processo de envelhecimento da pessoa idosa com dependência em seu bem-estar subjetivo. Metodologia: Pesquisa de natureza qualitativa. Os dados foram coletados através de entrevista semiestruturada aos idosos e ao gestor de um centro-dia municipal e através de diário de campo das atividades desenvolvidas pelo Centro Dia. As entrevistas foram transcritas e analisadas através da análise de discurso utilizando o referencial teórico do bem-estar subjetivo e satisfação com a vida. Resultados: Foram entrevistados 21 idosos de ambos os sexos com idade entre 63 e 94 anos. Através da leitura flutuante emergiram as seguintes categorias: As percepções sobre a velhice; O significado do Centro Dia; Investimento na rotina e no bem-estar subjetivo; Manutenção de vínculos afetivos e familiares; Desenvolvimento em novas aprendizagens; Manutenção de sentido de vida. Segundo os idosos os cuidados oferecidos pelo Centro Dia, proporcionam a construção de novos relacionamentos sociais, oportunidade de aprendizado, desenvolvimento de criatividade e de atividades que estimulam a autoestima, inclusive proporciona um novo sentido de vida. Lá eles recebem cuidados especializados, sendo que o serviço se mantem vigilante sobre suas condições de saúde, o que lhes traz segurança. Entretanto, alguns idosos referem frequentar o serviço por falta de opção pois devido às suas condições sociais e de saúde, não podem ficar desacompanhados em seus domicílios como desejam. Os cuidados estão organizados em atividades terapêuticas e de ocupação do tempo. Mensalmente são realizadas reuniões com os familiares para relatar as atividades desenvolvidas e a evolução de seu familiar, esclarecer dúvidas, oferecer orientações para o cuidado no domicílio. Conclusões: Envelhecer envolve múltiplos fatores incluindo individuais, psicológicos, biológicos e sociais. A velhice pode ser percebida como uma etapa bem-sucedida da vida. Desta forma, o bem-estar subjetivo percebido pelos usuários do centro dia foi identificado como importante componente que permite possibilidades de adaptação às mudanças advindas do envelhecimento e de condições associadas com vistas à promoção da velhice com qualidade de vida.

The significant increase in longevity in the Brazilian reality had as one of its consequences, the growth in the number of dependent people who need long-term care. This care is usually provided by family members and, due to changes in family structure and composition and the lack of preparation to provide this care, it ends up causing physical, financial and emotional overload to those caregivers. The Day-Care Center for the elderly is an instrument that is characterized as a space for reception, protection and coexistence of dependent elderly people whose families are unable to provide care during the whole day or part of it, also offering support and guidance to family members on the care of their elderly. So far, there are few studies on the perception of the subjective well-being of users of these services offered by the municipal public network. Objective: Analyze the contribution of the care offered by Day Care Center to the aging process of the dependent elderly person on their subjective well-being. Methodology: Qualitative research. Data were collected through semi-structured interviews with the elderly and the manager of a municipal day center and through a field diary of the activities carried out by the Day Care Center. The interviews were transcribed and analyzed through discourse analysis using the theoretical framework of subjective well-being and life satisfaction. Results: 21 elderly people of both sexes between 63 and 94 years old were interviewed. Through floating reading, the following categories emerged: Perceptions about old age; The meaning of the Day Care Center: Investment in routine and subjective well-being; Maintenance of affective and family bonds; Development of new learning; Maintenance of meaning of life. Results: According to the elderly, the care offered by Day Care Center provides the building of new social relationships, an opportunity for learning, the development of creativity and activities that encourage self-esteem, including providing a new meaning for life. They receive specialized care, and the service remains vigilant about their health conditions, which gives them security. However, some elderly people report attending the service due to lack of options because, in view of their social and health conditions, they cannot stay in their homes unattended as they would prefer. Care is organized into therapeutic and time-occupation activities. Monthly meetings are held with family members to report the activities developed and the evolution of their family member, answer questions, offer guidelines for care at home. Conclusions: Aging involves multiple factors including individual, psychological, biological and social ones. Old age can be perceived as a successful stage of life. This way, perceived subjective well-being by day care center users was identified as an important component that allows possibilities of adaptation to changes arising from aging and associated conditions in order to promote old age with quality of life.

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