Eugenia Carvalho - Academia.edu (original) (raw)

Papers by Eugenia Carvalho

Biomolecules, Feb 12, 2024

This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY

European Journal of Clinical Investigation

IntroductionDiabetes mellitus (DM) impairs wound healing. The aim was to determine whether DM inf... more IntroductionDiabetes mellitus (DM) impairs wound healing. The aim was to determine whether DM influences mitochondrial respiration in wounded skin (WS) and non‐wounded skin (NWS), in a pre‐clinical wound healing model of streptozotocin (STZ)‐induced diabetes.MethodsSix weeks after diabetes induction, two wounds were created in the back of C57BL/J6 mice. Using high‐resolution respirometry (HRR), oxygen flux was measured, in WS and NWS, using two substrate‐uncoupler‐inhibitor titration protocols, at baseline (day 0), day 3 and 10 post‐wounding, in STZ‐DM and non‐diabetic (NDM) mice. Flux control ratios for the oxidative phosphorylation (OXPHOS) capacity were calculated.ResultsA significant increase in mitochondrial respiration was observed in STZ‐DM skin compared to control skin at baseline. The OXPHOS capacity was decreased in WS under diabetes at day 3 post‐wounding (inflammation phase). However, at day 10 post‐wounding (remodeling phase), the OXPHOS capacity was higher in WS from S...

Diabetologia, Sep 1, 2021

Biochimica Et Biophysica Acta: Molecular Basis Of Disease, Jun 1, 2018

Background: Cardiovascular diseases (CVDs) are leading cause of death and primary cause of morbid... more Background: Cardiovascular diseases (CVDs) are leading cause of death and primary cause of morbidity and mortality in diabetic population. Epicardial adipose tissue (EAT) covers the heart's surface and is a source of biomolecules regulating heart and blood vessel physiology. The protective activation of the unfolded protein response (UPR) and autophagy allows the cardiomyocyte reticular network to restore energy and/or nutrient homeostasis and to avoid cell death. However, an excessive or prolonged UPR activation can trigger cell death. UPR activation is an early event of diabetic cardiomyopathies and deregulated autophagy is associated with CVDs. Results: An upregulation of UPR markers (glucose-regulated protein 78KDa, glucose-regulated protein 94KDa, inositol-requiring enzyme 1α, protein kinase RNAlike ER kinase and CCAAT/-enhancer-binding protein homologous protein (CHOP) gene) in EAT compared to subcutaneous adipose tissue (SAT), was observed as well as the UPR-related apoptosis marker caspase-4/procaspase-4 ratio but not in CHOP protein levels. Additionally, levels of ubiquitin and ubiquitinated proteins were decreased in EAT. Moreover, upregulation of autophagy markers (5' adenosine monophosphate-activated protein kinase, mechanistic target of rapamycin, Beclin 1, microtubule-associated protein light chain 3-II, lysosome-associated membrane protein 2, and PTEN-induced putative kinase 1) was observed, as well as an increase in the apoptotic Bim but not the ratio between Bim and the anti-apoptotic Bcl-2 in EAT. Diabetic patients show alterations in UPR activation markers but not in autophagy or apoptosis markers. Conclusion: UPR and autophagy are increased in EAT compared to SAT, opening doors to the identification of early biomarkers for cardiomyopathies and novel therapeutic targets.

Diabetology

Glucose, fructose, and galactose are widely used in the food industry as sweeteners and food addi... more Glucose, fructose, and galactose are widely used in the food industry as sweeteners and food additives. The over-consumption of these carbohydrates has been identified as a possible trigger of non-communicable diseases. These include insulin resistance, obesity, and type 2 diabetes. These sugars induce an energy overload with consequent adipose tissue (AT) expansion, contributing to the development of obesity. Furthermore, a common feature of these non-communicable diseases is the detrimental, chronic, low-grade inflammation contributing to their onset. In the present review, we identify the most widely used dietary free sugars and their direct impacts on AT metabolism and inflammation, as well as their involvement in systemic inflammation and effects on the immune cell phenotype and function. Additionally, we discuss the capacity of the free sugars to induce immune modulation, enhancing inflammation, an underlying hallmark of insulin resistance, obesity, and T2DM. Dietary sugars ha...

Diabetologia, Sep 1, 2019

Frontiers in Medicine

PurposeTear fluid biomarkers may offer a non-invasive strategy for detecting diabetic patients wi... more PurposeTear fluid biomarkers may offer a non-invasive strategy for detecting diabetic patients with increased risk of developing diabetic retinopathy (DR) or increased disease progression, thus helping both improving diagnostic accuracy and understanding the pathophysiology of the disease. Here, we assessed the tear fluid of nondiabetic individuals, diabetic patients with no DR, and diabetic patients with nonproliferative DR (NPDR) or with proliferative DR (PDR) to find putative biomarkers for the diagnosis and staging of DR.MethodsTear fluid samples were collected using Schirmer test strips from a cohort with 12 controls and 54 Type 2 Diabetes (T2D) patients, and then analyzed using mass spectrometry (MS)-based shotgun proteomics and bead-based multiplex assay. Tear fluid-derived small extracellular vesicles (EVs) were analyzed by transmission electron microscopy, Western Blotting, and nano tracking.ResultsProteomics analysis revealed that among the 682 reliably quantified proteins...

As the knowledge base and importance of mitochondrial physiology to evolution, health and disease... more As the knowledge base and importance of mitochondrial physiology to evolution, health and disease expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow the latest SI guidelines and those of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols with the nomenclature of classical bioenergetics. We endeavour to provide a balanced view of mitochondrial respirat...

Aging Cell, 2020

Age‐associated mitochondrial dysfunction and oxidative damage are primary causes for multiple hea... more Age‐associated mitochondrial dysfunction and oxidative damage are primary causes for multiple health problems including sarcopenia and cardiovascular disease (CVD). Though the role of Nrf2, a transcription factor that regulates cytoprotective gene expression, in myopathy remains poorly defined, it has shown beneficial properties in both sarcopenia and CVD. Sulforaphane (SFN), a natural compound Nrf2‐related activator of cytoprotective genes, provides protection in several disease states including CVD and is in various stages of clinical trials, from cancer prevention to reducing insulin resistance. This study aimed to determine whether SFN may prevent age‐related loss of function in the heart and skeletal muscle. Cohorts of 2‐month‐old and 21‐ to 22‐month‐old mice were administered regular rodent diet or diet supplemented with SFN for 12 weeks. At the completion of the study, skeletal muscle and heart function, mitochondrial function, and Nrf2 activity were measured. Our studies revealed a significant drop in Nrf2 activity and mitochondrial functions, together with a loss of skeletal muscle and cardiac function in the old control mice compared to the younger age group. In the old mice, SFN restored Nrf2 activity, mitochondrial function, cardiac function, exercise capacity, glucose tolerance, and activation/differentiation of skeletal muscle satellite cells. Our results suggest that the age‐associated decline in Nrf2 signaling activity and the associated mitochondrial dysfunction might be implicated in the development of age‐related disease processes. Therefore, the restoration of Nrf2 activity and endogenous cytoprotective mechanisms by SFN may be a safe and effective strategy to protect against muscle and heart dysfunction due to aging.

Pharmacological Research, 2020

Please cite this article as: da Silva Figueiredo AM, Leal EC, de Carvalho EML, Protein tyrosine p... more Please cite this article as: da Silva Figueiredo AM, Leal EC, de Carvalho EML, Protein tyrosine phosphatase 1B inhibition as a potential therapeutic target for chronic wounds in diabetes,

Diabetes, 2020

Diabetic neuropathy plays an important role in the development of chronic wounds. Endothelial pro... more Diabetic neuropathy plays an important role in the development of chronic wounds. Endothelial progenitor cells (EPCs) are known important regulators of wound angiogenesis. However, the role of neuropeptides in EPC function, and particularly Neurotensin (NT), is unknown in diabetic wound healing. EPCs isolated from normal mice were cultured in high glucose (25 mM) or normoglicemia (5.5 mM) and treated with NT (25-400 nM) for 3 days. Moreover, EPCs isolated from streptozotocin (STZ)-induced diabetic mice were treated with NT (100 nM) for 3 days. Subsequently, EPC viability, cell adhesion and reactive oxygen species (ROS) levels were evaluated. To the in vivo wound healing model, 6 mm wounds were created in the dorsum of diabetic mice and wound progression was followed up to day 10. The wound edges were injected with ex-vivo diabetic bone marrow derived EPCs treated in culture with NT (100 nM) for 3 days, or EPCs without NT treatment, or vehicle. Angiogenesis was evaluated by immunohis...

Life Sciences, 2020

This is a PDF file of an article that has undergone enhancements after acceptance, such as the ad... more This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Journal of Peptide Science, 2018

Neurotensin, substance P, and insulin have been demonstrated to improve wound healing in vivo. Ho... more Neurotensin, substance P, and insulin have been demonstrated to improve wound healing in vivo. However, the mechanism behind their effect is still not fully understood. This study investigates the effects leading to enhanced scratch closure by these peptides in vitro. The skin keratinocyte cell line, HaCaT, was used to test scratch closure effects of the peptides and alterations of cytokine levels. HUVEC cells were used to test the angiogenic effect of the peptides. Furthermore, clinical isolates of Staphylococcus lugdunensis were used to examine the potential antimicrobial activity of each peptide. Our results demonstrate that neurotensin, substance P, and insulin had significant migratory effects in scratch assays were neurotensin had the lowest effect. Furthermore, we investigated use of the peptides in combination. When substance P was used in combination with neurotensin, the cell migratory capacity was decreased, and the peptides showed a negative correlation (r = −0.298, P &l...

Clinical Immunology, 2019

Immune systems have evolved to recognize and eliminate pathogens and damaged cells. In humans, it... more Immune systems have evolved to recognize and eliminate pathogens and damaged cells. In humans, it is estimated to recognize 10 9 epitopes and natural selection ensures that clonally expanded cells replace unstimulated cells and overall immune cell numbers remain stationary. But, with age, it faces continuous repertoire restriction and concomitant accumulation of primed cells. Changes shaping the aging immune system have bitter consequences because, as inflammatory responses gain intensity and duration, tissue-damaging immunity and inflammatory disease arise. During inflammation, the glycolytic flux cannot cope with increasing ATP demands, limiting the immune response's extent. In diabetes, higher glucose availability stretches the glycolytic limit, dysregulating proteostasis and increasing T-cell expansion. Long-term hyperglycemia exerts an accumulating effect, leading to higher inflammatory cytokine levels and increased cytotoxic mediator secretion upon infection, a phenomenon known as diabetic chronic inflammation. Here we review the etiology of diabetic chronic inflammation and its consequences on wound healing.

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2018

This manuscript on 'Mitochondrial respiratory states and rates' is a position statement in the fr... more This manuscript on 'Mitochondrial respiratory states and rates' is a position statement in the frame of COST Action CA15203 MitoEAGLE. The list of co-authors evolved beyond phase 1 (phase 1 versions 1-44) in the bottom-up spirit of COST. This is an open invitation to scientists and students to join as co-authors, to provide a balanced view on mitochondrial respiratory control, a fundamental introductory presentation of the concept of the protonmotive force, and a consensus statement on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Phase 2: MitoEAGLE preprint 'The protonmotive force and respiratory control' (Versions 01-21): We continue to invite comments and suggestions, particularly if you are an early career investigator adding an open future-oriented perspective, or an established scientist providing a balanced historical basis. Your critical input into the quality of the manuscript will be most welcome, improving our aims to be educational, general, consensusoriented, and practically helpful for students working in mitochondrial respiratory physiology.

Archives of Dermatological Research, 2016

Diabetic foot ulcers affect 15-20 % of all diabetic patients and remain an important challenge si... more Diabetic foot ulcers affect 15-20 % of all diabetic patients and remain an important challenge since the available therapies have limited efficacy and some of the novel therapeutic approaches, which include growth factors and stem cells, are highly expensive and their safety remains to be evaluated. Despite its low cost and safety, the interest for topical insulin as a healing agent has increased only in the last 20 years. The molecular mechanisms of insulin signaling and its metabolic effects have been well studied in its classical target tissues. However, little is known about the specific effects of insulin in healthy or even diabetic skin. In addition, the mechanisms involved in the effects of insulin on wound healing have been virtually unknown until about 10 years ago. This paper will review the most recent advances in the cellular and molecular mechanisms that underlie the beneficial effects of insulin on skin wound healing in diabetes. Emerging evidence that links dysfunction of key cellular organelles, namely the endoplasmic reticulum and the mitochondria, to changes in the autophagy response, as well as the impaired wound healing in diabetic patients will also be discussed along with the putative mechanisms whereby insulin could regulate/modulate these alterations.

Journal of physiology and pharmacology : an official journal of the Polish Physiological Society, 2015

Side-effect minimization strategies to avoid serious side-effects of cyclosporine A (CsA), such a... more Side-effect minimization strategies to avoid serious side-effects of cyclosporine A (CsA), such as nephrotoxicity, have been mainly based on dose reduction and conversion to other putatively less nephrotoxic drugs, such as sirolimus (SRL), an inhibitor of the mammalian target of rapamycin. This study intended to evaluate the impact of protocols based on CsA dose reduction and further conversion to SRL on kidney function and lesions, based on serum, urine and renal tissue markers. The following 3 groups (n=6) were tested during a 9-week protocol: control (vehicle); CsA (5 mg/kg/day) and Red + Conv (CsA 30 mg/kg/day during 3 weeks + 3 weeks with CsA 5 mg/kg/day + SRL 1 mg/kg/day during the last 3 weeks). The following parameters were analysed: blood pressure, heart rate and biochemical data; serum and urine contents and clearances of creatinine, urea and neutrophil gelatinase-associated lipocalin (NGAL), as well as, glomerular filtration rate; kidney lipid peroxidation and clearance; ...