LUCIA DE NORONHA
@pucpr.br
ESCOLA DE MEDICINA E CIÊNCIAS DA VIDA
PONTIFICIA UNIVERSIDADDE CATOLICA DO PARANÁ
RESEARCH, TEACHING, or OTHER INTERESTS
Histology, Pathology and Forensic Medicine, Immunology and Allergy, Infectious Diseases
Scopus Publications
Scopus Publications
Suelen Teixeira Scheifer, Arieli Carini Michels, Filipe Modolo, Emanuela Carla dos Santos, Rafaela Scariot, Maria Cassia Ferreira Aguiar, Sergio Aparecido Ignácio, Lucia de Noronha, Laysa Toschi Martins, Henrique Ravanhol Frigeri,et al.
Elsevier BV
Liziane C. M. da Silva, Andressa C. dos Santos Maia, Nágila C. F. de Sousa, Catielen P. Pavi, Beatriz P. Savi, Seigo Nagashima, Samara Damasceno, Ayda H. Schneider, Lucas Z. Mascarin, João F. S. Rodrigues,et al.
MDPI AG
Chikungunya virus (CHIKV), the causative agent of the chikungunya fever, is an alphavirus widely transmitted by the bite of the female mosquito of the genus Aedes sp., especially in tropical and subtropical regions. Brazil is the country most affected by the microorganism. CHIKV classically induces articular pain, which can become long lasting for even years in a great number of the infected individuals, reducing their quality of life. The mechanisms of CHIKV-induced pain are poorly understood, but recent evidence indicated a role for the transient receptor potential vanilloid 1 (TRPV1) in this pathology. Herein, we assessed the ability of intra-articularly injected inactivated CHIKV or its RNA to trigger nociception in mice. Both stimuli induced bilateral secondary hyperalgesia to mechanical and heat stimuli. These responses were attenuated by TRPV1 ablation or antagonism. Joint structural alterations and increased cartilage TRPV1 protein expression were detected in the ipsilateral knee joints injected with either CHIKV or viral RNA. However, the lack of this receptor did not influence the histological changes triggered by CHIKV or RNA. The results further support the role of TRPV1 in CHIKV-induced pain and highlight its importance in the chronic phase of the disease.
Aldini Beuting Pereira, Arieli Carini Michels, Sarah Freygang Mendes Pilati, Filipe Modolo, Ana Paula Camargo Martins, Caroline Busatta Vaz De Paula, Seigo Nagashima, Lúcia Noronha, Ana Clara Prado Fonseca, Ariane Jéssica Torres Turmina,et al.
Public Library of Science (PLoS)
Objective The objective was to evaluate the early-stage immunoexpression of markers (COX-2, NF-kB, VEGFR-1 and apoptotic index) related to inflammation, angiogenesis, and cell survival in the tongue dorsum epithelium of mice exposed to hookah smoke. Materials and methods The sample consisted of Swiss mice (N = 20), female gender, aged 2 months, and approximately 25g each, four groups (n = 5) mice: group exposed to fresh air and groups exposed to hookah smoke for 7, 15, and 30 days. Tongues were embedded in paraffin. A tissue microarray was constructed, and immunohistochemistry was performed for Cyclooxygenase 2, NF-kappa B, Vascular Endothelial Growth Factor Receptor 1, and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) for apoptotic analysis. The positive and negative cells were quantified in the epithelium of the mid-dorsal tongue region. Kruskal-Wallis and Dunn tests was made. Results The apoptotic index was higher at 30 days of smoke exposure (20.38% basal/ 19.63%/ suprabasal) compared to the group exposed to air (9.55%/ 11.88%), The expression of Vascular Endothelial Growth Factor Receptor 1 was higher at 30 days of smoke exposure (30.15%/ 38.15%) compared to the group exposed to air (18.25%/ 3.60%). Conclusion Hookah smoke induced greater apoptosis and increased expression of Vascular Endothelial Growth Factor Receptor 1 in the epithelium of the tongue at 30 days, potentially playing a role in the initial stages of carcinogenesis, in the early stages of hookah use.
Raíssa Campos D’Amico, Seigo Nagashima, Lucas Baena Carstens, Karina de Guadalupe Bertoldi, Sabrina Mataruco, Júlio Cesar Honório D’Agostini, Elisa Carolina Hlatchuk, Sofia Brunoro da Silva, Lucia de Noronha, and Cristina Pellegrino Baena
MDPI AG
Obesity has been identified as an independent risk factor for severe COVID-19 unfavorable outcomes. Several factors, such as increased ACE2 receptor expression and chronic inflammation, can contribute to this relationship, yet the activation of the NLRP3 inflammasome pathway is also a key element. Our primary goal was to determine whether chronic NLRP3 inflammasome activation in people with obesity is different in critical COVID-19 and in critical chronic conditions. A retrospective analysis was conducted using clinical data and post-mortem lung tissue samples from 14 COVID-19 patients with obesity (group A) and 9 patients with obesity who died from non-COVID-19 causes (group B). Immunohistochemical analysis assessed twelve markers related to the NLRP3 inflammasome pathway. Group A showed a significantly higher expression of ASC (p = 0.0387) and CASP-1 (p = 0.0142). No significant differences were found for IL-8, TNF-α, NF-kB, NLRP3, IL-1β, and gasdermin-D. Group B had higher levels of IL-6 (p < 0.0001), IL-18 (p = 0.002), CASP-9 (p < 0.0001), and HIF (p = 0.0327). We concluded that COVID-19 activates the NLRP3 inflammasome pathway, possibly leading to pyroptotic cell death mediated by caspase-1. In contrast, people with obesity without COVID-19, despite exhibiting some markers of the NLRP3 inflammasome, are more likely to experience necroptosis mediated by caspase-9.
Guilherme Vieira Cavalcante, Rosangela Fedato, Lucia de Noronha, Seigo Nagashima, Ana Paula Camargo Martins, Márcia Olandoski, Ricardo Pinho, Aline Takejima, Rossana Simeoni, Julio Cesar Francisco,et al.
MDPI AG
Achilles tendon injuries are extremely common and have a significant impact on the physical and mental health of individuals. Both conservative and surgical treatments have unsatisfactory results. The search for new therapeutic tools, using cell therapies with stem cells (SC) and biological tissues, such as amniotic membranes (AM), has proved useful for the regeneration of injured tendons. Background/Objectives: This research was carried out to assess the capacity of tissue repair in animal models of Achilles tendinopathy, in which rats were submitted to complete sections of the tendon, and the effects of using bone marrow SC and/or AM graft are evaluated. Methods: Thirty-seven Wistar rats, submitted to complete surgical section of the Achilles tendon and subsequent tenorrhaphy, were randomized into four groups: Control Group (CG), received saline solution; SC Group (SCG) received an injection of SC infiltrated directly into the tendon; AM Group (AMG), the tendon was covered with an AM graft; SC + AM Group (SC+AMG), has been treated with an AM graft and SC local injection. Six weeks later, the Achilles tendons were evaluated using a histological score and immunohistochemical pro-healing markers such as Interferon-γ, AKT, and mTOR. Results: There were no differences between morphometric histological when evaluating the Achilles tendons of the samples. No significant differences were found regarding the expression of AKT-2 and mTOR markers between the study groups. The main finding was the presence of a higher concentration of Interferon-γ in the group treated with SC and AM. Conclusions: The isolated use of SC, AM, or the combination of SC-AM did not produce significant changes in tendon healing when the histological score was evaluated. Similarly, no difference was observed in the expression of AKT-2 and mTOR markers. An increase in the expression of Interferon-γ was observed in SC+AMG. This suggests that such therapies may be potentially beneficial for the regeneration of injured tendons. However, as tendon repair mechanisms are very complex, further studies should be carried out to verify the benefits of the tendon structure and function.
Aline L. Takejima, Rossana B. Simeoni, Milka L. Takejima, Angela G. Lemke, Seigo Nagashima, Anna C. F. Silva, Julio C. Francisco, Ricardo A. Pinho, Lúcia de Noronha, and Luiz C. Guarita-Souza
Informa UK Limited
Several studies have focused on novel therapeutic strategies for extensive skin lesions aiming to improve healing quality and reduce treatment duration. In this context, the use of amniotic membrane (AM) and Wharton's jelly (WJ) emerges as promising alternatives. Full-thickness dorsal skin wounds were created in 21 Wistar rats, randomly divided into three groups: control (C), AM - covered by AM and WJ - covered by WJ. Wound contraction rate was measured weekly. On day 28, histochemical (Picrosirius red) and immunohistochemical analyses matrix metalloproteinase-9 (MMP-9), transforming growth factor beta (TGF-β), and alpha-smooth muscle actin (α-SMA) were performed. On day seven, wound contraction rate was higher in the AM group (38.8%), followed by the WJ (27.4%) and control (26.5%) with statistically significance. During the first 14 days, the AM group maintained the highest contraction rate, followed by the control and WJ groups. However, by day 21, wound contraction rates increased in the order of WJ to AM to control groups (85.6%, 87.0%, and 91.1%) with statistically significance. Type I collagen predominated across all groups, without statistically significant differences among them. TGF-β expression significantly increased from WJ to AM to control groups (19.75%, 26.00%, and 36.56%) with statistically significance. MMP-9 and α-SMA expressions decreased from control to WJ to AM groups, but no significant differences were observed. Both AM and WJ enhanced early wound contraction and may support skin repair by attenuating fibrotic signaling. These findings highlight the potential of AM and WJ as biomaterials for promoting tissue regeneration at epithelial barriers.
Paulo Andre Bispo Machado-Junior, Andre Lass, Julia de Bortolo, Leticia Bressan Anizelli, Mateus T. Rocha, Henrique Machado Sousa Proença, Stephanie Rubianne Silva Carvalhal, Samya Hamad Mehanna, Seigo Nagashima, Luiz Claudio Fernandes,et al.
American Chemical Society (ACS)
Ana Paula Andreolla, Alessandra Abel Borges, Seigo Nagashima, Caroline Busatta Vaz de Paula, Lucia de Noronha, Nilson I.T. Zanchin, Juliano Bordignon, and Claudia Nunes Duarte dos Santos
Springer Science and Business Media LLC
AbstractOrthobunyavirus oropouche ense virus (OROV), the causative agent of Oropouche fever, is widely dispersed in Brazil and South America, causing sporadic outbreaks. Due to the similarity of initial clinical symptoms caused by OROV with other arboviruses found in overlapping geographical areas, differential diagnosis is challenging. As for most neglected tropical diseases, there is a shortage of reagents for diagnosing and studying OROV pathogenesis. We therefore developed and characterized mouse monoclonal antibodies and, one of them recognizes the OROV nucleocapsid in indirect immunofluorescent (IFA) and immunohistochemistry (IHC) assays. Considering that it is the first monoclonal antibody produced for detecting OROV infections, we believe that it will be useful not only for diagnostic purposes but also for performing serological surveys and epidemiological surveillance on the dispersion and prevalence of OROV in Brazil and South America.
Marina de Castro Deus, Ana Carolina Gadotti, Erika Sousa Dias, Júlia Bacarin Monte Alegre, Beatriz Akemi Kondo Van Spitzenbergen, Gabriela Bohnen Andrade, Sara Soares Tozoni, Rebecca Benicio Stocco, Marcia Olandoski, Felipe Francisco Bondan Tuon,et al.
MDPI AG
COVID-19 is characterized by pronounced hypercytokinemia. The cytokine switch, marked by an imbalance between pro-inflammatory and anti-inflammatory cytokines, emerged as a focal point of investigation throughout the COVID-19 pandemic. However, the kinetics and temporal dynamics of cytokine release remain contradictory, making the development of new therapeutics difficult, especially in severe cases. This study collected serum samples from SARS-CoV-2 infected patients at 72 h intervals and monitored them for various cytokines at each timepoint until hospital discharge or death. Cytokine levels were analyzed based on time since symptom onset and patient outcomes. All cytokines studied prospectively were strong predictors of mortality, particularly IL-4 (AUC = 0.98) and IL-1β (AUC = 0.96). First-timepoint evaluations showed elevated cytokine levels in the mortality group (p < 0.001). Interestingly, IFN-γ levels decreased over time in the death group but increased in the survival group. Patients who died exhibited sustained levels of IL-1β and IL-4 and increased IL-6 levels over time. These findings suggest cytokine elevation is crucial in predicting COVID-19 mortality. The dynamic interplay between IFN-γ and IL-4 highlights the balance between Th1/Th2 immune responses and underscores IFN-γ as a powerful indicator of immune dysregulation throughout the infection.
Gisela H Pontes, Clara P W Ramos, Lucia de Noronha, Fernando Serra-Guimarães, Amanda S Cavalcanti, Ana Paula F Barbosa, and Maria Eugenia L Duarte
Oxford University Press (OUP)
Abstract Background Polyurethane (PU)-coated breast implants are known for their strong integration into breast tissue and the formation of capsules around them. However, capsular contracture can pose both aesthetic and clinical challenges. Objectives The objectives of this study were to analyze the biological and morphological characteristics of the capsular tissue surrounding PU-coated implants, irrespective of their contracture status, and to assess their potential suitability as a flap in revisional breast surgery for capsular contracture. Methods A total of 23 tissue samples were harvested from the capsules surrounding PU-coated breast implants in 12 female patients during replacement or revisional surgery. We evaluated collagen abundance, cellular and vascular density, inflammation, collagen band types and alignment, synovial metaplasia, capsule thickness, and the expression of inflammatory biomarkers and myofibroblasts with immunohistochemical techniques. Scanning electron microscopy was employed to assess implant surface characteristics over time. Results We found a significant association of capsule contraction with longer implantation durations and greater implant surface roughness (P = .018 and P = .033, respectively). Synovial metaplasia was significantly more frequent in noncontracted capsules (P = .0049). Both capsule types consisted of paucicellular, type I collagen-rich compact fibrous tissue with low vascularization. There was a marked reduction in inflammatory cells within the foreign body granuloma. The expression of inflammatory biomarkers in the capsular tissue was negligible. Conclusions Given the reduced levels of inflammatory and vascular components within the dense, fibrous capsular tissue, we consider them to be viable alternatives for capsular flaps in revisional surgery. This strategy has the potential to mimic the reconstruction achieved with acellular dermal matrix. Level of Evidence: 4
Maiara Carolina Perussolo, Bassam Felipe Mogharbel, Cláudia Sayuri Saçaki, Nádia Nascimento da Rosa, Ana Carolina Irioda, Nathalia Barth de Oliveira, Julia Maurer Appel, Larissa Lührs, Leanderson Franco Meira, Luiz Cesar Guarita-Souza,et al.
MDPI AG
This study aims to evaluate and compare cellular therapy with human Wharton’s jelly (WJ) mesenchymal stem cells (MSCs) and neural precursors (NPs) in experimental autoimmune encephalomyelitis (EAE), a preclinical model of Multiple Sclerosis. MSCs were isolated from WJ by an explant technique, differentiated to NPs, and characterized by cytometry and immunocytochemistry analysis after ethical approval. Forty-eight rats were EAE-induced by myelin basic protein and Freund’s complete adjuvant. Forty-eight hours later, the animals received intraperitoneal injections of 250 ng/dose of Bordetella pertussis toxin. Fourteen days later, the animals were divided into the following groups: a. non-induced, induced: b. Sham, c. WJ-MSCs, d. NPs, and e. WJ-MSCs plus NPs. 1 × 105. Moreover, the cells were placed in a 10 µL solution and injected via a stereotaxic intracerebral ventricular injection. After ten days, the histopathological analysis for H&E, Luxol, interleukins, and CD4/CD8 was carried out. Statistical analyses demonstrated a higher frequency of clinical manifestation in the Sham group (15.66%) than in the other groups; less demyelination was seen in the treated groups than the Sham group (WJ-MSCs, p = 0.016; NPs, p = 0.010; WJ-MSCs + NPs, p = 0.000), and a lower cellular death rate was seen in the treated groups compared with the Sham group. A CD4/CD8 ratio of <1 showed no association with microglial activation (p = 0.366), astrocytes (p = 0.247), and cell death (p = 0.577) in WJ-MSCs. WJ-MSCs and NPs were immunomodulatory and neuroprotective in cellular therapy, which would be translated as an adjunct in demyelinating diseases.
Maria Riedmeier, Shipra Agarwal, Sonir Antonini, Tatiana E I Jaick B Costa, Orhan Diclehan, Martin Fassnacht, Bonald C Figueiredo, Tulay Guran, Christoph Härtel, Imme Haubitz,et al.
Oxford University Press (OUP)
Abstract Objective Pediatric adrenocortical carcinoma (pACC) is rare, and prognostic stratification remains challenging. We aimed to confirm the prognostic value of the previously published pediatric scoring system (pS-GRAS) in an international multicenter cohort. Design Analysis of pS-GRAS items of pACC from 6 countries in collaboration of ENSAT-PACT, GPOH-MET, and IC-PACT. Methods We received patient data of the pS-GRAS items including survival information from 9 centers. PS-GRAS score was calculated as a sum of tumor stage (1 = 0; 2-3 = 1; 4 = 2 points), grade (Ki67 index: 0%-9% = 0; 10%-19% = 1; ≥20% = 2 points), resection status (R0 = 0; RX/R1/R2 = 1 point), age (&lt;4 years = 0; ≥4 years = 1 point), and hormone production (androgen production = 0; glucocorticoid-/mixed-/no-hormone production = 1 point) generating 8 scores and 4 groups (1: 0-2, 2: 3-4, 3: 5, 4: 6-7). Primary endpoint was overall survival (OS). Results We included 268 patients with median age of 4 years. The analysis of the pS-GRAS score showed a significantly favorable prognosis in patients with a lower scoring compared to higher scoring groups (5-year OS: Group 1 98%; group 2 87% [hazard ratio {HR} of death 3.6, 95% CI of HR 1.6-8.2]; group 3 43% [HR of death 2.8, 95% CI 1.9-4.4]; group 4: OS 18% [HR of death 2.1, 95% CI 1.7-2.7]). In the multivariable analysis, age (HR of death 3.5, 95% CI 1.8-7.0), resection status (HR of death 5.5, 95% CI 2.7-11.1), tumor stage (HR of death 1.9, 95% CI of HR 1.2-3.0), and Ki67 index (HR of death 1.7, 95% CI 1.2-2.4) remained strong independent outcome predictors. Especially infants &lt; 4 years showed more often low-risk constellations with a better OS for all tumor stages. Conclusion In an international multicenter study, we confirmed that the pS-GRAS score is strongly associated with overall survival among patients with pACC. Age, resection status, stage, and Ki67 index are important parameters for risk stratification.
Aloysio Enck Neto, Katia Martins Foltz, Thiago Fuchs, Luize Kremer Gamba, Marcos Antonio Denk, Paulo Cesar Lock Silveira, Thatyanne Gradowski do Nascimento, Alice Machado Clemencia, Julio César Francisco, Lucia de Noronha,et al.
MDPI AG
Background: Tracheal grafts have been investigated for over a century, aiming to replace various lesions. However, tracheal reconstruction surgery remains a challenge, primarily due to anatomical considerations, intraoperative airway management, the technical complexity of reconstruction, and the potential postoperative morbidity and mortality. Due to research development, the amniotic membrane (AM) and Wharton’s Jelly (WJ) arise as alternatives within the new set of therapeutic alternatives. These structures hold significant therapeutic potential for tracheal defects. This study analyzed the capacity of tracheal tissue regeneration after 60 days of decellularized WJ and AM implantation in rabbits submitted to conventional tracheostomy. Methods: An in vivo experimental study was carried out using thirty rabbits separated into three groups (Control, AM, and WJ) (n = 10). The analyses were performed 60 days after surgery through immunohistochemistry. Results: Different immunomarkers related to scar regeneration, such as aggrecan, TGF-β1, and α-SMA, were analyzed. However, they highlighted no significant difference between the groups. Collagen type I, III, and Aggrecan also showed no significant difference between the groups. Conclusions: Both scaffolds appeared to be excellent frameworks for tissue engineering, presenting biocompatibility and a desirable microenvironment for cell survival; however, they did not display histopathological benefits in trachea tissue regeneration.
Chin Wee Tan, Jinjin Chen, Ning Liu, Dharmesh D. Bhuva, Tony Blick, James Monkman, Caroline Cooper, Malvika Kharbanda, Kristen Feher, Belinda Phipson,et al.
Elsevier BV
Claudia Martins Galindo, Letícia Milani, Lucas Trevisan França de Lima, Eliana Rezende Adami, Simei Go, Lucia de Noronha, Olair Carlos Beltrame, Giseli Klassen, Edneia Amancio de Souza Ramos, Ronald P.J. Oude Elferink,et al.
Elsevier BV
Arieli Carini Michels, Suelen Teixeira Scheifer, Fernanda Tiboni, Filipe Modolo, Emanuela Carla dos Santos, Rafaela Scariot, Maria Cassia Ferreira Aguiar, Sergio Aparecido Ignácio, Lucia de Noronha, Mariana Hornung Marins,et al.
Springer Science and Business Media LLC
ALINE L. TAKEJIMA, PAULO ANDRÉ B. MACHADO-JÚNIOR, GUSTAVO G. BLUME, ROSSANA BAGGIO SIMEONI, JULIO CESAR FRANCISCO, MURILO S. TONIAL, LUIS FELIPE B. MARQUEZE, LUCIA NORONHA, MARCIA OLANDOSKI, ELTYEB ABDELWAHID,et al.
FapUNIFESP (SciELO)
Recent studies have suggested that therapies with stem cells and amniotic membrane can modulate the inflammation following an ischemic injury in the heart. This study evaluated the effects of bone-marrow mononuclear cells (BMMC) and acellular human amniotic membrane (AHAM) on cardiac function and NLRP3 complex in a rat model of heart failure.On the 30th day,the echocardiographic showed improvements on ejection fraction and decreased pathological ventricular remodeling on BMMC and AHAM groups.Oxidative stress analysis was similar between the three groups,and the NLRP3 inflammasome activity were not decreased with the therapeutic use of both BMMC and AHAM,in comparison to the control group.
Nataly Stylianou, Ismail Sebina, Nicholas Matigian, James Monkman, Hadeel Doehler, Joan Röhl, Mark Allenby, Andy Nam, Liuliu Pan, Anja Rockstroh,et al.
Wiley
AbstractObjectivesSevere Acute Respiratory Syndrome Coronavirus 2 (SARS‐CoV‐2) virus infection in pregnancy is associated with higher incidence of placental dysfunction, referred to by a few studies as a ‘preeclampsia‐like syndrome’. However, the mechanisms underpinning SARS‐CoV‐2‐induced placental malfunction are still unclear. Here, we investigated whether the transcriptional architecture of the placenta is altered in response to SARS‐CoV‐2 infection.MethodsWe utilised whole‐transcriptome, digital spatial profiling, to examine gene expression patterns in placental tissues from participants who contracted SARS‐CoV‐2 in the third trimester of their pregnancy (n = 7) and those collected prior to the start of the coronavirus disease 2019 (COVID‐19) pandemic (n = 9).ResultsThrough comprehensive spatial transcriptomic analyses of the trophoblast and villous core stromal cell subpopulations in the placenta, we identified SARS‐CoV‐2 to promote signatures associated with hypoxia and placental dysfunction. Notably, genes associated with vasodilation (NOS3), oxidative stress (GDF15, CRH) and preeclampsia (FLT1, EGFR, KISS1, PAPPA2) were enriched with SARS‐CoV‐2. Pathways related to increased nutrient uptake, vascular tension, hypertension and inflammation were also enriched in SARS‐CoV‐2 samples compared to uninfected controls.ConclusionsOur findings demonstrate the utility of spatially resolved transcriptomic analysis in defining the underlying pathogenic mechanisms of SARS‐CoV‐2 in pregnancy, particularly its role in placental dysfunction. Furthermore, this study highlights the significance of digital spatial profiling in mapping the intricate crosstalk between trophoblasts and villous core stromal cells, thus shedding light on pathways associated with placental dysfunction in pregnancies with SARS‐CoV‐2 infection.
Rogil Jose de Almeida Torres, Fernando Moreto, Andrea Luchini, Rogerio Joao de Almeida Torres, Sofia Pimentel Longo, Ricardo Aurino Pinho, Seigo Nagashima, Lucia de Noronha, Artur Junio Togneri Ferron, Carol Cristina Vagula de Almeida Silva,et al.
Springer Science and Business Media LLC
Abstract Background To assess oxidative effects induced by a high-calorie diet on the retina of Wistar rats and test the antioxidative effects of carnosine supplementation. Methods Wistar rats were randomly divided into the following groups: standard diet (SD), high-calorie diet (HcD), standard diet + carnosine (SD + Car), and high-calorie diet + carnosine (HcD + Car). The body weight, adiposity index, plasma glucose, total lipids, high-density lipoprotein (HDL), low-density lipoprotein (LDL), uric acid, creatinine, and triglycerides of the animals were evaluated. The retinas were analyzed for markers of oxidative stress. Hydrogen peroxide production was assessed by 2',7'-dichlorodihydrofluorescein diacetate (DCF) oxidation. The total glutathione (tGSH), total antioxidant capacity (TAC), protein carbonyl, and sulfhydryl groups of the antioxidant system were analyzed. Results TAC levels increased in the retinas of the SD + Car group compared to the SD group (p < 0.05) and in the HcD + Car group compared to the HcD group (p < 0.05). The levels of GSH and the GSSH:GSSG ratio were increased in the HcD + Car group compared to the SD + Car group (p < 0.05). An increase in the retinal carbonyl content was observed in the HcD group compared to the SD group (p < 0.05) and in the HcD + Car group compared to the SD + Car group (p < 0.05). A high-calorie diet (HcD) was also associated with a decrease in retinal sulfhydryl-type levels compared to the SD group (p < 0.05). Conclusion The results suggest that feeding a high-calorie diet to rats can promote an increase in carbonyl content and a reduction in sulfhydryl groups in their retinas. The administration of carnosine was not effective in attenuating these oxidative markers. Trial registration Animal Ethics Committee of Botucatu Medical School - Certificate number 1292/2019.
Xiao Tan, Laura F. Grice, Minh Tran, Onkar Mulay, James Monkman, Tony Blick, Tuan Vo, Ana Clara Almeida, Jarbas da Silva Motta, Karen Fernandes de Moura,et al.
Wiley
AbstractThe SARS‐CoV‐2 (COVID‐19) virus has caused a devastating global pandemic of respiratory illness. To understand viral pathogenesis, methods are available for studying dissociated cells in blood, nasal samples, bronchoalveolar lavage fluid and similar, but a robust platform for deep tissue characterization of molecular and cellular responses to virus infection in the lungs is still lacking. We developed an innovative spatial multi‐omics platform to investigate COVID‐19‐infected lung tissues. Five tissue‐profiling technologies were combined by a novel computational mapping methodology to comprehensively characterize and compare the transcriptome and targeted proteome of virus infected and uninfected tissues. By integrating spatial transcriptomics data (Visium, GeoMx and RNAScope) and proteomics data (CODEX and PhenoImager HT) at different cellular resolutions across lung tissues, we found strong evidence for macrophage infiltration and defined the broader microenvironment surrounding these cells. By comparing infected and uninfected samples, we found an increase in cytokine signalling and interferon responses at different sites in the lung and showed spatial heterogeneity in the expression level of these pathways. These data demonstrate that integrative spatial multi‐omics platforms can be broadly applied to gain a deeper understanding of viral effects on cellular environments at the site of infection and to increase our understanding of the impact of SARS‐CoV‐2 on the lungs.
Gabriela M. Coelho, Allan H. D. Cataneo, Sonia M. Raboni, Meri B. Nogueira, Caroline B. Vaz de Paula, Ana C. S. F. Almeida, Vanessa Z. Rogerio, Nilson T. Zanchin, Lucia de Noronha, Camila Zanluca,et al.
Wiley
AbstractSince its emergence in late 2019, coronavirus disease 2019 (COVID‐19) has caused millions of deaths and socioeconomic losses. Although vaccination significantly reduced disease mortality, it has been shown that protection wanes over time, and that severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) variants of concern (VOCs) may escape vaccine‐derived immunity. Therefore, serological studies are necessary to assess protection in the population and guide vaccine regimens. A common measure of protective immunity is the presence of neutralizing antibodies (nAbs). However, the gold standard for measuring nAbs (plaque reduction neutralization test, or PRNT) is laborious and time‐consuming, limiting its large‐scale applicability. We developed a high‐throughput fluorescence reduction neutralization assay (FRNA) to detect SARS‐CoV‐2 nAbs. Because the assay relies on immunostaining, we developed and characterized monoclonal antibodies (mAbs) to lower costs and reduce the assay's vulnerability to reagent shortages. Using samples of individuals vaccinated with COVID‐19 and unvaccinated/pre‐pandemic samples, we showed that FRNA results using commercial and in‐house mAbs strongly correlated with those of the PRNT method while providing results in 70% less time. In addition to providing a fast, reliable, and high‐throughput alternative for measuring nAbs, the FRNA can be easily customized to assess SARS‐CoV‐2 VOCs. Additionally, the mAb we produced was able to detect SARS‐CoV‐2 in pulmonary tissues by immunohistochemistry assays.
Ana Paula Dergham, Caroline Busatta Vaz de Paula, Seigo Nagashima, Márcia Olandoski, Lucia de Noronha, and Vanessa Santos Sotomaior
MDPI AG
Advanced high-grade serous ovarian carcinoma is a serious malignant neoplasm with a late diagnosis and high mortality rate. Even when treated with standard therapy, such as surgery followed by carboplatin and paclitaxel chemotherapy, the prognosis remains unfavorable. Immunotherapy is a treatment alternative that requires further study. Therefore, we aimed to evaluate the expression of PD-1, PD-L1, CD8, MSI (MLH1, MSH2, MSH6, and PMS2), and p53 in the paraffin samples of high-grade serous ovarian carcinoma. A retrospective study of 28 southern Brazilian patients with advanced serous ovarian carcinoma (EC III or IV) was conducted between 2009 and 2020. The expression of these proteins was evaluated using immunohistochemistry, and the results were correlated with the patients’ clinicopathological data. At diagnosis, the mean age was 61 years, and the most common clinical stage (60%) was EC III. Among the cases, 84.6% exhibited p53 overexpression, 14.8% had MSI, 92.0% were sensitive to platinum, and more than 50.0% relapsed after treatment. Patients with MSI had a lower CD8/PD-1 ratio and more relapses (p = 0.03). In conclusion, analysis of immunotherapeutic markers in paraffin-embedded advanced serous ovarian carcinoma samples is feasible and may assist in prognosis.
Isio Carvalho de Souza, Aline Luri Takejima, Rossana Baggio Simeoni, Luize Kremer Gamba, Victoria Stadler Tasca Ribeiro, Katia Martins Foltz, Lucia de Noronha, Meila Bastos de Almeida, Jose Rocha Faria Neto, Katherine Athayde Teixeira de Carvalho,et al.
MDPI AG
Wound healing is a complex process of repair that involves the interaction between different cell types and involves coordinated interactions between intracellular and extracellular signaling. Bone Marrow Mesenchymal Stem Cells (BMSCs) based and acellular amniotic membrane (AM) therapeutic strategies with the potential for treatment and regeneration of tissue. We aimed to evaluate the involvement of paracrine effects in tissue repair after the flap skin lesion rat model. In the full-thickness flap skin experiment of forty Wistar rats: A total of 40 male Wistar rats were randomized into four groups: group I: control (C; n = 10), with full-thickness lesions on the back, without (BMSCs) or AM (n = 10); group II: injected (BMSCs; n = 10); group III: covered by AM; group IV–injected (AM + BMSCs; n = 10). Cytokine levels, IL-1, and IL-10 assay kits, superoxide dismutase (SOD), glutathione reductase (GRs) and carbonyl activity levels were measured by ELISA 28th day, and TGF-β was evaluated by immunohistochemical, the expression collagen expression was evaluated by Picrosirius staining. Our results showed that the IL-1 interleukin was higher in the control group, and the IL-10 presented a higher mean when compared to the control group. The groups with BMSCs and AM showed the lowest expression levels of TGF-β. SOD, GRs, and carbonyl activity analysis showed a predominance in groups that received treatment from 80%. The collagen fiber type I was predominant in all groups; however, the AM + BMSCs group obtained a higher average when compared to the control group. Our findings suggest that the AM+ BMSCs promote skin wound healing, probably owing to their paracrine effect attributed to the promotion of new collagen for tissue repair.
Anna Flavia Ribeiro Santos Miggiolaro, Felipe Paes Gomes da Silva, David Batista Wiedmer, Thiago Mateus Godoy, Nicolas Henrique Borges, Giulia Werner Piper, Alessandro G. G. Oricil, Carolline Konzen Klein, Elisa Carolina Hlatchuk, Júlio César H. Dagostini,et al.
MDPI AG
COVID-19 has been considered a vascular disease, and inflammation, intravascular coagulation, and consequent thrombosis may be associated with endothelial dysfunction. These changes, in addition to hypoxia, may be responsible for pathological angiogenesis. This research investigated the impact of COVID-19 on vascular function by analyzing post-mortem lung samples from 24 COVID-19 patients, 10 H1N1pdm09 patients, and 11 controls. We evaluated, through the immunohistochemistry technique, the tissue immunoexpressions of biomarkers involved in endothelial dysfunction, microthrombosis, and angiogenesis (ICAM-1, ANGPT-2, and IL-6, IL-1β, vWF, PAI-1, CTNNB-1, GJA-1, VEGF, VEGFR-1, NF-kB, TNF-α and HIF-1α), along with the histopathological presence of microthrombosis, endothelial activation, and vascular layer hypertrophy. Clinical data from patients were also observed. The results showed that COVID-19 was associated with increased immunoexpression of biomarkers involved in endothelial dysfunction, microthrombosis, and angiogenesis compared to the H1N1 and CONTROL groups. Microthrombosis and vascular layer hypertrophy were found to be more prevalent in COVID-19 patients. This study concluded that immunothrombosis and angiogenesis might play a key role in COVID-19 progression and outcome, particularly in patients who die from the disease.
Arutha Kulasinghe, Ning Liu, Chin Wee Tan, James Monkman, Jane E. Sinclair, Dharmesh D. Bhuva, David Godbolt, Liuliu Pan, Andy Nam, Habib Sadeghirad,et al.
Wiley
AbstractThe severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is known to present with pulmonary and extra‐pulmonary organ complications. In comparison with the 2009 pandemic (pH1N1), SARS‐CoV‐2 infection is likely to lead to more severe disease, with multi‐organ effects, including cardiovascular disease. SARS‐CoV‐2 has been associated with acute and long‐term cardiovascular disease, but the molecular changes that govern this remain unknown. In this study, we investigated the host transcriptome landscape of cardiac tissues collected at rapid autopsy from seven SARS‐CoV‐2, two pH1N1, and six control patients using targeted spatial transcriptomics approaches. Although SARS‐CoV‐2 was not detected in cardiac tissue, host transcriptomics showed upregulation of genes associated with DNA damage and repair, heat shock, and M1‐like macrophage infiltration in the cardiac tissues of COVID‐19 patients. The DNA damage present in the SARS‐CoV‐2 patient samples, were further confirmed by γ‐H2Ax immunohistochemistry. In comparison, pH1N1 showed upregulation of interferon‐stimulated genes, in particular interferon and complement pathways, when compared with COVID‐19 patients. These data demonstrate the emergence of distinct transcriptomic profiles in cardiac tissues of SARS‐CoV‐2 and pH1N1 influenza infection supporting the need for a greater understanding of the effects on extra‐pulmonary organs, including the cardiovascular system of COVID‐19 patients, to delineate the immunopathobiology of SARS‐CoV‐2 infection, and long term impact on health.