Effect of Blueberry Supplementation on a Diet-Induced Rat Model of Prediabetes-Focus on Hepatic Lipid Deposition, Endoplasmic Stress Response and Autophagy.

Blueberries, red fruits enriched in polyphenols and fibers, are envisaged as a promising nutraceutical intervention in a plethora of metabolic diseases. Prediabetes, an intermediate state between normal glucose tolerance and type 2 diabetes, fuels the development of complications, including hepatic steatosis. In previous work, we have demonstrated that blueberry juice (BJ) supplementation benefits glycemic control and lipid profile, which was accompanied by an amelioration of hepatic mitochondrial bioenergetics. The purpose of this study is to clarify the impact of long-term BJ nutraceutical intervention on cellular mechanisms that govern hepatic lipid homeostasis, namely autophagy and endoplasmic reticulum (ER) stress, in a rat model of prediabetes. Two groups of male Wistar rats, 8-weeks old, were fed a prediabetes-inducing high-fat diet (HFD) and one group was fed a control diet (CD). From the timepoint where the prediabetic phenotype was achieved (week 16) until the end of the study (week 24), one of the HFD-fed groups was daily orally supplemented with 25 g/kg body weight (BW) of BJ (HFD + BJ). BW, caloric intake, glucose tolerance and insulin sensitivity were monitored throughout the study. The serum and hepatic lipid contents were quantified. Liver and interscapular brown and epidydimal white adipose tissue depots (iBAT and eWAT) were collected for histological analysis and to assess thermogenesis, ER stress and autophagy markers. The gut microbiota composition and the short-chain fatty acids (SCFAs) content were determined in colon fecal samples. BJ supplementation positively impacted glycemic control but was unable to prevent obesity and adiposity. BJ-treated animals presented a reduction in fecal SCFAs, increased markers of arrested iBAT thermogenesis and energy expenditure, together with an aggravation of HFD-induced lipotoxicity and hepatic steatosis, which were accompanied by the inhibition of autophagy and ER stress responses in the liver. In conclusion, despite the improvement of glucose tolerance, BJ supplementation promoted a major impact on lipid management mechanisms at liver and AT levels in prediabetic animals, which might affect disease course.

An Overview of Biosimilars-Development, Quality, Regulatory Issues, and Management in Healthcare.

Biological therapies have transformed high-burden treatments. As the patent and exclusivity period for biological medicines draws to a close, there is a possibility for the development and authorization of biosimilars. These products boast comparable levels of safety, quality, and effectiveness to their precursor reference products. Biosimilars, although similar to reference products, are not identical copies and should not be considered generic substitutes for the original. Their development and evaluation involve a rigorous step-by-step process that includes analytical, functional, and nonclinical evaluations and clinical trials. Clinical studies conducted for biosimilars aim to establish similar efficacy, safety, and immunogenicity, rather than demonstrating a clinical benefit, as with the reference product. However, although the current knowledge regarding biosimilars has significantly increased, several controversies and misconceptions still exist regarding their immunogenicity, extrapolation, interchangeability, substitution, and nomenclature. The development of biosimilars stimulates market competition, contributes toward healthcare sustainability, and allows for greater patient access. However, maximizing the benefits of biosimilars requires cooperation between regulators and developers to ensure that patients can benefit quickly from access to these new therapeutic alternatives while maintaining high standards of quality, safety, and efficacy. Recognizing the inherent complexities of comprehending biosimilars fully, it is essential to focus on realistic approaches, such as fostering open communication between healthcare providers and patients, encouraging informed decision-making, and minimizing risks. This review addresses the regulatory and manufacturing requirements for biosimilars and provides clinicians with relevant insights for informed prescribing.

Addiction interprofessional experiences of care: Stress, coping and transformation.

This study aimed to explore healthcare professionals' attribution of meaning to the concept of addiction, treatment and recovery. The study was conducted through semi-structured interviews with 18 healthcare professionals in the field of addiction: nine nurses, six psychologists, a psychiatrist, a social worker and a psychosocial technician. Based on a qualitative methodology, a thematic analysis was performed using the NVivo software. Regarding professional stress, the results revealed several stress factors related to the intervention (e.g. strategic powerlessness and uncertainty of intervention's effectiveness), the addicted individuals (e.g. motivation, difficulties in adhering to treatment, patient behaviour, negative patient emotionality and lack of socio-economic resources) as well as the healthcare services (e.g. team stress). Adaptive coping strategies were reported, both used in the work context (e.g. cognitive coping strategies and acceptance) and in personal life (e.g. work-family boundaries and family support and closeness). Despite the professional stress experienced, the participants' narratives indicated positive transformations of the self, which suggests resilient professional trajectories.

Antisenescence Therapy Improves Function in a Human Model of Cardiac Fibrosis-on-a-Chip.

Cardiac fibrosis is a significant contributor to heart failure and is characterized by abnormal ECM deposition and impaired contractile function. We have previously developed a model of cardiac fibrosis via TGF-ß treatment of engineered microtissues using heart-on-a-chip technology which incorporates human induced pluripotent stem cell-derived cardiomyocytes and cardiac fibroblasts. Here, we describe that these cardiac fibrotic tissues expressed markers associated with cellular senescence via transcriptomic analysis. Treatment of fibrotic tissues with the senolytic drugs dasatinib and quercetin (D+Q) led to an improvement of contractile function, reduced passive tension, and downregulated senescence-related gene expression, an outcome we were previously unable to achieve using standard-of-care drugs. The improvement in functional parameters was also associated with a reduction in fibroblast density, though no changes in absolute collagen deposition were observed. This study demonstrates the benefit of senolytic treatment for cardiac fibrosis in a human-relevant model, supporting data in animal models, and will enable the further elucidation of cell-specific effects of senolytics and how they impact cardiac fibrosis and senescence.

CK2ß Regulates Hematopoietic Stem Cell Biology and Erythropoiesis.

The Ser-Thr kinase CK2 plays important roles in sustaining cell survival and resistance to stress and these functions are exploited by different types of blood tumors. Yet, the physiological involvement of CK2 in normal blood cell development is poorly known. Here, we discovered that the ß regulatory subunit of CK2 is critical for normal hematopoiesis in the mouse. Fetal livers of conditional CK2ß knockout embryos showed increased numbers of hematopoietic stem cells associated to a higher proliferation rate compared to control animals. Both hematopoietic stem and progenitor cells (HSPCs) displayed alterations in the expression of transcription factors involved in cell quiescence, self-renewal, and lineage commitment. HSPCs lacking CK2ß were functionally impaired in supporting both in vitro and in vivo hematopoiesis as demonstrated by transplantation assays. Furthermore, KO mice developed anemia due to a reduced number of mature erythroid cells. This compartment was characterized by dysplasia, proliferative defects at early precursor stage, and apoptosis at late-stage erythroblasts. Erythroid cells exhibited a marked compromise of signaling cascades downstream of the cKit and erythropoietin receptor, with a defective activation of ERK/JNK, JAK/STAT5, and PI3K/AKT pathways and perturbations of several transcriptional programs as demonstrated by RNA-Seq analysis. Moreover, we unraveled an unforeseen molecular mechanism whereby CK2 sustains GATA1 stability and transcriptional proficiency. Thus, our work demonstrates new and crucial functions of CK2 in HSPC biology and in erythropoiesis.

Transcriptome Analysis Identifies the Crosstalk between Dendritic and Natural Killer Cells in Human Cutaneous Leishmaniasis.

Skin ulcers of cutaneous leishmaniasis (CL) are characterized by a localized inflammatory response mediated by innate and adaptive immune cells, including dendritic cells (DC) and natural killer (NK) cells. Bidirectional interactions between DCs and NK cells contribute to tailor leishmaniasis outcome. Despite advances in the Leishmania biology field in recent decades, the mechanisms involved in DC/NK-mediated control of Leishmania sp. pathogenesis as well as the cellular and molecular players involved in such interaction remain unclear. The present study sought to investigate canonical pathways associated with CL arising from Leishmania braziliensis infection. Initially, two publicly available microarray datasets of skin biopsies from active CL lesions were analyzed, and five pathways were identified using differentially expressed genes. The "Crosstalk between DCs and NK cells" pathway was notable due to a high number of modulated genes. The molecules significantly involved in this pathway were identified, and our findings were validated in newly obtained CL biopsies. We found increased expression of TLR4, TNFRSF1B, IL-15, IL-6, CD40, CCR7, TNF and IFNG, confirming the analysis of publicly available datasets. These findings reveal the "crosstalk between DCs and NK cells" as a potential pathway to be further explored in the pathogenesis of CL, especially the expression of CCR7, which is correlated with lesion development.

Adhesive Capsulitis' Patients Report Improvement in Functionality Trough International Classification of Functioning, Disability and Health Checklist and Dash After Suprascapular Nerve Blocks.

Objective To evaluate the functionality in patients with adhesive capsulitis undergoing suprascapular nerve block (SSNB). Methods A before-and-after clinical prospective study in a single center was conducted with patients with secondary adhesive capsulitis treated with four nerve blocks based on anatomical limits. The sample was non-probabilistic, and it was obtained after a routine appointment at a specialized outpatient clinic. The instruments used for evaluation were the International Classification of Functioning, Disability and Health (ICF) and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire, which were applied at baseline (T 0 ), one week after the fourth SSNB (T 4 ), and three months after the first SSNB (T 12 ). The paired t -test was used to compare the means of the ICF checklist items and DASH in the different: T 0 xT 4 ; T 4 xT 12 ; and T 0 xT 12 ). The probability of rejecting the null hypothesis was 5%. Results The sample was composed of 25 individuals with a mean age of 58.16 years; 16 of them were female. The duration of the pain symptoms ranged from 2 to 16 months, with a mean of 5.92 months. The ICF checklist showed that all domains had already improved in T 4, except for the environmental factors, which only improved at 03 months ( p = 0.037). The patients reported improvements in shoulder function in T 4 , which increased more in T 12 , at the end of data collection ( p = 0.019). Conclusion The SSNB technique is effective in patients with adhesive capsulitis after 4 weeks of application, with improvements in individual's functionality lasting for 12 weeks.

Adhesive Capsulitis' Patients Report Improvement in Functionality Trough International Classification of Functioning, Disability and Health Checklist and Dash After Suprascapular Nerve Blocks / Pacientes com capsulite adesiva relatam melhora da funcionalidade pela Classificação Internacional de Funcionalidade, Incapacidade e Saúde e DASH após bloqueios do nervo supraescapular

Abstract Objective To evaluate the functionality in patients with adhesive capsulitis undergoing suprascapular nerve block (SSNB). Methods A before-and-after clinical prospective study in a single center was conducted with patients with secondary adhesive capsulitis treated with four nerve blocks based on anatomical limits. The sample was non-probabilistic, and it was obtained after a routine appointment at a specialized outpatient clinic. The instruments used for evaluation were the International Classification of Functioning, Disability and Health (ICF) and the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire, which were applied at baseline (T0), one week after the fourth SSNB (T4), and three months after the first SSNB (T12). The paired t-test was used to compare the means of the ICF checklist items and DASH in the different: T0xT4;T4xT12;and T0xT12). The probability of rejecting the null hypothesis was 5%. Results The sample was composed of 25 individuals with a mean age of 58.16 years; 16 of them were female. The duration of the pain symptoms ranged from 2 to 16 months, with a mean of 5.92 months. The ICF checklist showed that all domains had already improved in T4 except for the environmental factors, which only improved at 03 months (p = 0.037). The patients reported improvements in shoulder function in T4, which increased more in T12, at the end of data collection (p = 0.019). Conclusion The SSNB technique is effective in patients with adhesive capsulitis after4 weeks of application, with improvements in individual's functionality lasting for 12 weeks.

Resumo Objetivo Avaliar a funcionalidade em pacientes com capsulite adesiva submetidos a bloqueio do nervo supraescapular (BNSE). Métodos Um estudo clínico prospectivo do tipo antes e depois foi realizado em um único centro com pacientes com capsulite adesiva secundária tratados com quatro bloqueios baseados em limites anatômicos. A amostra foi não probabilística, tendo sido obtida após consulta de rotina em ambulatório especializado. Os instrumentos utilizados para avaliação foram a Classificação Internacional de Funcionalidade, Incapacidade e Saúde (CIF) e o questionário de Disfunções do Braço, Ombro e Mão (DASH), que foram aplicados antes da intervenção (T0), uma semana após o quarto BNSE (T4),etrêsapósoprimeiroBNSE(T12).AsmédiasdositensdaCIFedoDASHnos diferentes tempos (T0 x T4; T4 x T12; e T0 x T12) foram comparadas por meio do teste t pareado. A probabilidade de rejeitar a hipótese nula foi de 5%. Resultados A amostra foi composta por 25 indivíduos com média de idade de 58,16 anos; 16 eram mulheres. A duração dos sintomas dolorosos variou de 2 a 16 meses, com média de 5,92 meses. A CIF mostrou que todos os domínios já haviam melhorado em T4 à exceção dos fatores ambientais, que só melhoraram aos 3 meses (p = 0,037). Os pacientes já relataram melhora na função do ombro em T4 emaisainda em T12,ao finaldacoletadedados (p = 0,019). Conclusão A técnica de BNSE é eficaz em pacientes com capsulite adesiva após 4 semanas de aplicação, com melhora da funcionalidade do indivíduo e sua manutenção até 12 semanas.

Steroid hormone levels and bone mineral density in women over 65 years of age.

Previous studies using immunoassays for steroid measurements have focused on the association between steroid hormone levels and bone mineral density (BMD) in postmenopausal women, obtaining contradictory results. This study aimed to assess this association using a highly sensitive bioanalytical method. A total of 68 postmenopausal women, aged 65-89 years, were enrolled in a cross-sectional study. Measurements of the BMD of the hip and lumbar spine were performed using dual energy X-ray absorptiometry, and serum hormone levels were quantified by gas chromatography and tandem mass spectrometry. Associations between estradiol (E2), testosterone, dehydroepiandrosterone (DHEA), androstenedione and T score levels of the hip and lumbar spine were evaluated, after adjustment for confounding variables. The analysis revealed a statistically significant association between testosterone and the T score of the hip (p = 0.035), but not that of the lumbar spine. No statistically significant associations were found between E2, DHEA, androstenedione and the T scores of the hip and the lumbar spine. Using a highly sensitive hormone assay method, our study identified a significant association between testosterone and BMD of the hip in women over 65 years of age, suggesting that lower testosterone increases the risk of osteoporosis.

Modeling Heart Diseases on a Chip: Advantages and Future Opportunities.

Heart disease is a significant burden on global health care systems and is a leading cause of death each year. To improve our understanding of heart disease, high quality disease models are needed. These will facilitate the discovery and development of new treatments for heart disease. Traditionally, researchers have relied on 2D monolayer systems or animal models of heart disease to elucidate pathophysiology and drug responses. Heart-on-a-chip (HOC) technology is an emerging field where cardiomyocytes among other cell types in the heart can be used to generate functional, beating cardiac microtissues that recapitulate many features of the human heart. HOC models are showing great promise as disease modeling platforms and are poised to serve as important tools in the drug development pipeline. By leveraging advances in human pluripotent stem cell-derived cardiomyocyte biology and microfabrication technology, diseased HOCs are highly tuneable and can be generated via different approaches such as: using cells with defined genetic backgrounds (patient-derived cells), adding small molecules, modifying the cells' environment, altering cell ratio/composition of microtissues, among others. HOCs have been used to faithfully model aspects of arrhythmia, fibrosis, infection, cardiomyopathies, and ischemia, to name a few. In this review, we highlight recent advances in disease modeling using HOC systems, describing instances where these models outperformed other models in terms of reproducing disease phenotypes and/or led to drug development.

Steroid hormone levels in postmenopausal hysterectomised women with and without ovarian conservation: the continuous endocrine function of the ovaries.

This study aims to clarify the effect of postmenopausal bilateral oophorectomy on plasma steroid hormone levels. Women who were submitted in the postmenopausal period to hysterectomy for uterine benign conditions were divided into two groups: 18 women had isolated hysterectomy and 11 had hysterectomy with bilateral salpingo-oophorectomy. In both groups serum hormone levels were quantified by solid phase extraction and gas chromatography and tandem mass spectrometry. Differences in dehydroepiandrosterone (DHEA), testosterone, androstenedione and oestradiol were determined in both groups. The analysis revealed lower steroid levels in the bilateral salpingo-oophorectomy group when compared to the isolated hysterectomy group with statistically significant differences found for DHEA (5.8 ± 3.2 vs. 9.4 ± 4.4 ng/mL; p = 0.019) and oestradiol (0.69 ± 0.4 vs. 1.48 ± 4.3 ng/mL; p = 0.007). The results are consistent with a significant endocrine activity of the postmenopausal ovary. The clinical consequences of these findings need to be clarified and postmenopausal prophylactic bilateral salpingo-oophorectomy re-evaluated.IMPACT STATEMENTWhat is already known on this subject? Although it is consensual that premenopausal prophylactic bilateral oophorectomy should not be performed because it has harmful effects on women's health, the evidence regarding the effects of postmenopausal prophylactic bilateral oophorectomy is scarce and this procedure continues to be a regular practice. Few studies have demonstrated that postmenopausal ovaries still have endocrine activity that may impact older women's health.What do the results of this study add? This is the first study to compare hormone levels of postmenopausal women based on their hysterectomy and oophorectomy status using GC-MS/MS, a highly sensitive bioanalytical assay for the measurement of steroid hormones. Previous studies relied on immunoassays and did not compare DHEA levels, which according to the intracrinology theory is a precursor for androgens and oestrogens. In this study, statistically significant lower levels of DHEA and oestradiol were found after postmenopausal bilateral salpingo-oophorectomy.What are the implications of these findings for clinical practice and/or further research? This is a pilot study that may lead to further investigation in this area to clarify the impact of the prophylactic removal of postmenopausal ovaries on older women's health and lead to changes in surgical procedures.

Negative effects of the COVID-19 pandemic on physical activity and binge eating disorder in Brazilian women: a cross-sectional study / Efeitos negativos da pandemia de COVID-19 na atividade física e no transtorno da compulsão alimentar periódica em mulheres brasileiras: um estudo transversal / Efectos negativos de la pandemia de COVID-19 sobre la actividad física y el trastorno por atracón en mujeres brasileñas: un estudio transversal

ABSTRACT This study aimed to identify changes in physical activity, sitting time, and binge eating disorder in Brazilian women during the COVID-19 pandemic. A cross-sectional and retrospective study was conducted online on 150 Brazilian women adults. The prevalence of binge eating and physically inactive women increased during the COVID-19 pandemic (p = < 0.001 and p = 0.01, respectively). Binge eating was associated with insufficient physical activity before (p = 0.01) and during the pandemic (p = 0.05). The physical activity time and sitting time showed no differences after adjustment for possible confounders (p > 0.05). In conclusion, the pandemic affected Brazilian women's lifestyle and eating behavior.

RESUMO Este estudo teve como objetivo identificar mudanças na atividade física, tempo sentado e transtorno da compulsão alimentar periódica em mulheres brasileiras durante a pandemia de COVID-19. Foi realizado um estudo transversal e retrospectivo por meio de um formulário on-line em 150 mulheres adultas brasileiras. A prevalência de compulsão alimentar e de mulheres fisicamente inativas aumentou durante a pandemia de COVID-19 (p = < 0,001 e p = 0,01, respectivamente). A compulsão alimentar foi associada à atividade física insuficiente antes (p = 0,01) e durante a pandemia (p = 0,05). O tempo de atividade física e o tempo sentado não apresentaram diferenças após ajuste para possíveis fatores de confusão (p > 0,05). Em conclusão, a pandemia afetou o estilo de vida e o comportamento alimentar das mulheres brasileiras.

RESUMEN El objetivo del estudio fue identificar cambios los cambios en la actividad física, el tiempo sentado y el trastorno por atracón en mujeres brasileñas durante la pandemia de COVID-19. Es un estudio transversal y retrospectivo que evaluó, a través de un formulario en línea en 150 mujeres brasileñas adultas. La prevalencia de atracones y de mujeres físicamente inactivas aumentó durante la pandemia de COVID-19 (p = < 0,001 y p = 0,01, respectivamente). Los atracones se asociaron con actividad física insuficiente antes (p = 0,01) y durante la pandemia (p = 0,05). El tiempo de actividad física y el tiempo sentado no difirieron después del ajuste por posibles factores de confusión (p > 0,05). En conclusión, la pandemia afectó el estilo de vida y el comportamiento alimentario de las mujeres brasileñas.

Isolation of ready-made rat microvessels and its applications in effective in vivo vascularization and in angiogenic studies in vitro.

Despite recent advances in the differentiation of human pluripotent stem cells into multiple cell types for application in replacement therapies, tissue vascularization remains a bottleneck for regenerative medicine. Fragments of primary microvessels (MVs) harvested from adipose tissue retain endothelialized lumens and perivascular cell coverage. We have used these MVs to support the survival and engraftment of transplanted human pluripotent stem cell-derived cardiomyocytes, pancreatic progenitors or primary human islets. MVs connect with host vessels, perfuse with blood and form a hierarchal vascular network in vivo after subcutaneous or intracardiac transplantation. MVs also display the ability to remodel and form stable vascular networks with long-term retention (>3.5 months). MVs can be cultured in 3D hydrogels in vitro, where they retain vessel shape and undergo angiogenic sprouting without the need for exogenous growth factor supplementation. Therefore, MVs offer a robust vascularization strategy for regenerative medicine approaches and a platform for angiogenic studies and drug testing in vitro. Here we describe in detail the protocol for: (1) the isolation of MVs from rat epididymal fat by limited collagenase digestion, followed by size-selective sieving; (2) the incorporation of MVs into 3D collagen hydrogels; (3) the in vitro culture of MVs in 3D gels for angiogenic studies; and (4) the in vivo transplantation of 3D hydrogels containing MVs into the mouse subcutis. The isolation procedure does not require highly specific equipment and can be performed in ~3 h by researchers with experience in rodent handling and cell culture.

A human model of arteriovenous malformation (AVM)-on-a-chip reproduces key disease hallmarks and enables drug testing in perfused human vessel networks.

Brain arteriovenous malformations (AVMs) are a disorder wherein abnormal, enlarged blood vessels connect arteries directly to veins, without an intervening capillary bed. AVMs are one of the leading causes of hemorrhagic stroke in children and young adults. Most human sporadic brain AVMs are associated with genetic activating mutations in the KRAS gene. Our goal was to develop an in vitro model that would allow for simultaneous morphological and functional phenotypic data capture in real time during AVM disease progression. By generating human endothelial cells harboring a clinically relevant mutation found in most human patients (activating mutations within the small GTPase KRAS) and seeding them in a dynamic microfluidic cell culture system that enables vessel formation and perfusion, we demonstrate that vessels formed by KRAS4AG12V mutant endothelial cells (ECs) were significantly wider and more leaky than vascular beds formed by wild-type ECs, recapitulating key structural and functional hallmarks of human AVM pathogenesis. Immunofluorescence staining revealed a breakdown of adherens junctions in mutant KRAS vessels, leading to increased vascular permeability, a hallmark of hemorrhagic stroke. Finally, pharmacological blockade of MEK kinase activity, but not PI3K inhibition, improved endothelial barrier function (decreased permeability) without affecting vessel diameter. Collectively, our studies describe the creation of human KRAS-dependent AVM-like vessels in vitro in a self-assembling microvessel platform that is amenable to phenotypic observation and drug delivery.

Prediabetes Induces More Severe Acute COVID-19 Associated With IL-6 Production Without Worsening Long-Term Symptoms.

Aims: Pre-existing conditions, such as age, hypertension, obesity, and diabetes, constitute known risk factors for severe COVID-19. However, the impact of prediabetes mellitus (PDM) on COVID-19 severity is less clear. This study aimed to evaluate the influence of PDM in the acute and long-term phases of COVID-19. Materials and methods: We compared inflammatory mediators, laboratory and clinical parameters and symptoms in COVID-19 patients with prediabetes (PDM) and without diabetes (NDM) during the acute phase of infection and at three months post-hospitalization. Results: Patients with PDM had longer hospital stays and required intensive care unit admission more frequently than NDM. Upon hospitalization, PDM patients exhibited higher serum levels of interleukin 6 (IL-6), which is related to reduced partial pressure of oxygen (PaO2) in arterial blood, oxygen saturation (SpO2) and increased COVID-19 severity. However, at three months after discharge, those with PDM did not exhibit significant alterations in laboratory parameters or residual symptoms; however, PDM was observed to influence the profile of reported symptoms. Conclusions: PDM seems to be associated with increased risk of severe COVID-19, as well as higher serum levels of IL-6, which may constitute a potential biomarker of severe COVID-19 risk in affected patients. Furthermore, while PDM correlated with more severe acute-phase COVID-19, no long-term worsening of sequelae was observed.

A three-dimensional human adipocyte model of fatty acid-induced obesity.

Obesity prevalence has reached pandemic proportions, leaving individuals at high risk for the development of diseases such as cancer and type 2 diabetes. In obesity, to accommodate excess lipid storage, adipocytes become hypertrophic, which is associated with an increased pro-inflammatory cytokine secretion and dysfunction of metabolic processes such as insulin signaling and lipolysis. Targeting adipocyte dysfunction is an important strategy to prevent the development of obesity-associated disease. However, it is unclear how accurately animal models reflect human biology, and the long-term culture of human hypertrophic adipocytes in anin vitro2D monolayer is challenging due to the buoyant nature of adipocytes. Here we describe the development of a human 3Din vitrodisease model that recapitulates hallmarks of obese adipocyte dysfunction. First, primary human adipose-derived mesenchymal stromal cells are embedded in hydrogel, and infiltrated into a thin cellulose scaffold. The thin microtissue profile allows for efficient assembly and image-based analysis. After adipocyte differentiation, the scaffold is stimulated with oleic or palmitic acid to mimic caloric overload. Using functional assays, we demonstrated that this treatment induced important obese adipocyte characteristics such as a larger lipid droplet size, increased basal lipolysis, insulin resistance and a change in macrophage gene expression through adipocyte-conditioned media. This 3D disease model mimics physiologically relevant hallmarks of obese adipocytes, to enable investigations into the mechanisms by which dysfunctional adipocytes contribute to disease.

Effects of social distancing caused by the COVID-19 pandemic on physical activity level, sitting time, and binge eating: a comparison between overweight/obese and normal-weight adults.

Purpose: To compare changes in physical activity level (PAL), sitting time (ST), and binge eating disorder (BED) in overweight/obese adults vs. those normal weight during social distancing caused by the COVID-19 pandemic. Methods: A cross-sectional and retrospective study was carried out with adults of both sexes, aged 18-60 years, which assessed, through an online form, the PAL, ST, and BED pre (PSD) and during social distancing (DSD) caused by the COVID-19 pandemic. The PAL and ST were assessed by the short version of the International Physical Activity Questionnaire and the assessment of BED by the Binge Eating Scale. Results: 323 responses were included in the analysis (194 normal weight and 129 overweight/obese). There was a decrease in PAL and an increase in the prevalence of BED in both groups; nevertheless, the overweight/obese group had a 62% chance of presenting lower PALs than normal-weight individuals (OR = 1.62; 95% CI 1.03-2.55) and showed a lower total weekly physical activity time during the pandemic (adjusted p = 0.05). In addition, overweight individuals were more likely to binge eat before (OR = 4.21; 95% CI 2.10-8.45) and during the pandemic (OR = 4.24; 95% CI 2.54-7.06) and showed a higher prevalence during social distancing. Conclusion: The social distancing caused by the COVID-19 pandemic changed exercise behaviors and increased the prevalence of binge eating in the general population. However, overweight/obese participants engaged in less total weekly physical activity and showed a higher prevalence of binge eating before and during social distancing. Level of evidence: Level III; analytical observational cohort study.

Microvascular Dysfunction in Skeletal Muscle Precedes Myocardial Vascular Changes in Diabetic Cardiomyopathy: Sex-Dependent Differences.

Aim: To uncover sex-related microvascular abnormalities that underlie the early presentation of reduced perfusion in leg skeletal muscle in a type II rat model of diabetic cardiomyopathy. Methods and Results: Diabetes was induced using a non-obese, diet-based, low-dose streptozotocin model in adult female (18 diabetic, 9 control) and male rats (29 diabetic, 11 control). Time-course monitoring over 12 months following diabetes induction was performed using echocardiography, treadmill exercise, photoacoustic imaging, flow-mediated dilation (FMD), histopathology, and immunohistochemistry. Diabetic rats maintained normal weights. Hypertension appeared late in both diabetic males (7 months) and females (10 months), while only diabetic males had elevated cholesterol (7 months). On echocardiography, all diabetic animals maintained normal ejection fraction and exhibited diastolic dysfunction, mild systolic dysfunction, and a slightly enlarged left ventricle. Exercise tolerance declined progressively and early in males (4 months), later in females (8 months); FMD showed lower baseline femoral arterial flow but unchanged reactivity in both sexes (5 months); and photoacoustic imaging showed lower tissue oxygen saturation in the legs of diabetic males (4 months) and diabetic females (10 months). Myocardial perfusion was normal in both sexes. Histopathology at the final timepoint of Month 10 (males) and Month 12 (females) revealed that myocardial microvasculature was normal in both vessel density and structure, thus explaining normal perfusion on imaging. However, leg muscle microvasculature exhibited perivascular smooth muscle thickening around small arterioles in diabetic females and around large arterioles in diabetic males, explaining the depressed readings on photoacoustic and FMD. Histology also confirmed the absence of commonly reported HFpEF markers, including microvessel rarefaction, myocardial fibrosis, and left ventricular hypertrophy. Conclusion: Exercise intolerance manifesting early in the progression of diabetic cardiomyopathy can be attributed to decreased perfusion to the leg skeletal muscle due to perivascular smooth muscle thickening around small arterioles in females and large arterioles in males. This microvascular abnormality was absent in the myocardium, where perfusion levels remained normal throughout the study. We conclude that although skeletal muscle microvascular dysfunction of the vasculature presents at different levels depending on sex, it consistently presents early in both sexes prior to overt cardiac changes such as rarefaction, fibrosis, or hypertrophy.

Prosocial Tendencies Measure Validation for Portuguese Late Adolescents

O Questionário de Tendências Prossociais (MTP) de Carlo e Randall (2002) é amplamente usado para avaliar tendências prossociais. Partindo do pressuposto de que existem diferenças no comportamento pró-social entre adolescentes e adultos, o presente estudo teve como objetivo validar a versão em português do MTP com uma amostra de adolescentes. Este instrumento avalia seis tipos diferentes de tendências prossociais: altruísta, anónima, emergência, emocional, complacente e pública. Desenhado para confirmar esta estrutura fatorial do MTP com adolescentes portugueses, este estudo recolheu as perceções de um total de 1313 adolescentes a frequentar o ensino secundário. A análise fatorial confirmatória corroborou a estrutura de seis fatores da validação original. Além disso, considerando o ano escolar e o género dos alunos, a análise de invariância foi executada e atestou a invariância configural, métrica (parcialmente para género) e escalar (parcialmente para ano escolar e género) do MTP. Estes resultados demonstram ainda que a versão portuguesa MTP é confiável, aumentando a utilidade deste instrumento para avaliar comportamentos prossociais na adolescência.

The Prosocial Tendencies Measure (PTM; Carlo & Randall, 2002) is widely used for assessing prosocial tendencies. Assuming that there are differences in prosocial behavior between adolescents and adults, the current study aimed to further validate the Portuguese version of the PTM for adolescents. This instrument assesses six different types of prosocial tendencies ­ altruism, anonymous, dire, emotional, compliant and public. Designed to confirm this structure with Portuguese adolescents, this study collected a total of 1313 high school students' prosocial behaviors perceptions. Confirmatory factor analysis corroborated the six-factor structure of the original validation. Furthermore, considering the students' school year and gender, invariance analysis was executed and attested the PTM's configural, metric (partially for gender) and scalar (partially for school year and gender) invariance. These findings further demonstrate that the Portuguese version of the PTM is reliable, enhancing the utility of this instrument to assess prosocial behaviors in adolescence.

GP2-enriched pancreatic progenitors give rise to functional beta cells in vivo and eliminate the risk of teratoma formation.

Human pluripotent stem cell (hPSC)-derived pancreatic progenitors (PPs) can be differentiated into beta-like cells in vitro and in vivo and therefore have therapeutic potential for type 1 diabetes (T1D) treatment. However, the purity of PPs varies across different hPSC lines, differentiation protocols, and laboratories. The uncommitted cells may give rise to non-pancreatic endodermal, mesodermal, or ectodermal derivatives in vivo, hampering the safety of hPSC-derived PPs for clinical applications and their differentiation efficiency in research settings. Recently, proteomics and transcriptomics analyses identified glycoprotein 2 (GP2) as a PP-specific cell surface marker. The GP2-enriched PPs generate higher percentages of beta-like cells in vitro, but their potential in vivo remains to be elucidated. Here, we demonstrate that the GP2-enriched-PPs give rise to all pancreatic cells in vivo, including functional beta-like cells. Remarkably, GP2 enrichment eliminates the risk of teratomas, which establishes GP2 sorting as an effective method for PP purification and safe pancreatic differentiation.