MARCELO ALVES FERREIRA
Projetos de Pesquisa
Unidades Organizacionais
Departamento de Patologia, Faculdade de Medicina
LIM/59 - Laboratório de Biologia Celular, Hospital das Clínicas, Faculdade de Medicina
LIM/59 - Laboratório de Biologia Celular, Hospital das Clínicas, Faculdade de Medicina
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conferenceObject Immune Responses Of CVID Patients To COVID-19 Vaccines(2023) MEDEIROS, Giuliana Xavier de; FERREIRA, Loisi de Carvalho Pereira; MAGAWA, Jhosiene Yukari; KURAMOTO, Andreia; SASAHARA, Greyce Luri; FERREIRA, Marcelo; BARROS, Myrthes Maragna Toledo; KALIL, Jorge; MARINHO, Ana Karolina Barreto Berselli; CUNHA-NETO, Edecio; SANTOS, Keity Souza; KOKRON, Cristina- Titanium dioxide induced inflammation in the small intestine(2012) NOGUEIRA, Carolina Maciel; AZEVEDO, Walter Mendes de; DAGLI, Maria Lucia Zaidan; TOMA, Sergio Hiroshi; LEITE, Andre Zonetti de Arruda; LORDELLO, Maria Laura; NISHITOKUKADO, Ieda; ORTIZ-AGOSTINHO, Carmen Lucia; DUARTE, Maria Irma Seixas; FERREIRA, Marcelo Alves; SIPAHI, Aytan MirandaAIM: To investigate the effects of titanium dioxide (TiO2) nanoparticles (NPTiO2) and microparticles (MPTiO2) on the inflammatory response in the small intestine of mice. METHODS: BI 57/6 male mice received distilled water suspensions containing TiO2 (100 mg/kg body weight) as NPTiO2 (66 nm), or MPTiO2 (260 nm) by gavage for 10 d, once a day; the control group received only distilled water. At the end of the treatment the duodenum, jejunum and ileum were extracted for assessment of cytokines, inflammatory cells and titanium content. The cytokines interleukin (IL)-1b, IL-4, IL-6, IL-8, IL-10, IL-12, IL-13, IL-17, IL-23, tumor necrosis factor-alpha (TNF-alpha), intracellular interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta) were evaluated by enzyme-linked immunosorbent assay in segments of jejunum and ileum (mucosa and underlying muscular tissue). CD4(+) and CD8(+) T cells, natural killer cells, and dendritic cells were evaluated in duodenum, jejunum and ileum samples fixed in 10% formalin by immunohistochemistry. The titanium content was determined by inductively coupled plasma atomic emission spectrometry. RESULTS: We found increased levels of T CD4(+) cells (cells/mm(2)) in duodenum: NP 1240 +/- 139.4, MP 1070 +/- 154.7 vs 458 +/- 50.39 (P < 0.01); jejunum: NP 908.4 +/- 130.3, MP 813.8 +/- 103.8 vs 526.6 +/- 61.43 (P < 0.05); and ileum: NP 818.60 +/- 123.0, MP 640.1 +/- 32.75 vs 466.9 +/- 22.4 (P < 0.05). In comparison to the control group, the groups receiving TiO2 showed a statistically significant increase in the levels of the inflammatory cytokines IL-12, IL-4, IL-23, TNF-alpha, IFN-gamma and TGF-beta. The cytokine production was more pronounced in the ileum (mean SE): IL-12: NP 33.98 +/- 11.76, MP 74.11 +/- 25.65 vs 19.06 +/- 3.92 (P < 0.05); IL-4: NP 17.36 +/- 9.96, MP 22.94 +/- 7.47 vs 2.19 +/- 0.65 (P < 0.05); IL-23: NP 157.20 +/- 75.80, MP 134.50 +/- 38.31 vs 22.34 +/- 5.81 (P < 0.05); TNF alpha: NP 3.71 +/- 1.33, MP 5.44 +/- 1.67 vs 0.99 +/- 019 (P < 0.05); IFN gamma: NP 15.85 +/- 9.99, MP 34.08 +/- 11.44 vs 2.81 +/- 0.69 (P < 0.05); and TGF-alpha: NP 780.70 +/- 318.50, MP 1409.00 +/- 502.20 vs 205.50 +/- 63.93 (P < 0.05). CONCLUSION: Our findings indicate that TiO2 particles induce a Th1-mediated inflammatory response in the small bowel in mice.
- Protoporphyrin fluorescence induced by methyl-ALA in skin healing(2013) GONDIM, Roberta M. F.; VIEIRA, Vinicius C. C.; VERAS, Mariana M.; FERREIRA, Marcelo A.; CALDINI, Elia Tamaso Espin Garcia; MUNOZ, Daniel Romero; BAPTISTA, Mauricio S.Background: Estimation of the time period that precedes an injury is critical in forensic medicine. However, there is no reliable method that can be used to evaluate the oldness of a lesion. The aim of this work is to develop a fluorimetric method that can be used to follow the aging process of lesions by applying methyl ALA (MAL) on wounds and by quantifying protoporphyrin IX (PPIX) fluorescence during the healing process. We also aim to understand the changes in PPIX fluorescence by establishing a correlation with histological evaluations during the heating process. Methods: Standardized linear wounds were made on the dorsum of 72 mice, which were divided in control (MAL -) and experimental (MAL +) groups. In vivo fluorescence spectra (FS) were collected from normal and wound skin sites of control and experimental groups, corresponding to four groups of FS spectra: (a) FS of skin wound after MAL (+/+); (b) FS of normal skin after MAL (-/+); (c) FS of skin wound without MAL (+/-) and (d) FS of normal skin without MAL (-/-). Animals were monitored periodically for 3 months and euthanized. Tissue specimens were processed for histological analysis using design-based stereological methods. Serial cross-sections were analyzed to evaluate the organization of the dermis and epidermis, collagen deposition and cellular proliferation. Results: FS of skin wound with MAL (+/+) showed an expressive intensity increase from the beginning of the experiment to the 34th day, with maximum fluorescence being observed on the similar to 11th day after wounding. There was preferential PPIX accumulation in healing sites as compared to adjacent normal skin (+/-) in the early stage of healing. Histological findings allowed correlation of the fluorescence increase mainly with cell proliferation. The drastic decrease in the FS intensity observed in the end of the healing process was correlated with the decrease in the proliferation rate as well as with the presence of new extracellular fibrous materials. Conclusions: In the mice wound-healing model tested here, it was possible to distinguish whether the injury was in early or advanced stages by using PPIX fluorescence induced by MAL. We conclude that this method is a promising approach to evaluate the age of skin wounding and we hope this work will stimulate human studies to allow this technique to become standardized in forensic medicine.
- Immunodominant antibody responses directed to SARS-CoV-2 hotspot mutation sites and risk of immune escape(2023) OLIVEIRA, Jamille Ramos; RUIZ, Cesar Manuel Remuzgo; MACHADO, Rafael Rahal Guaragna; MAGAWA, Jhosiene Yukari; DAHER, Isabela Pazotti; URBANSKI, Alysson Henrique; SCHMITZ, Gabriela Justamante Haendel; ARCURI, Helen Andrade; FERREIRA, Marcelo Alves; SASAHARA, Greyce Luri; MEDEIROS, Giuliana Xavier de; JR, Roberto Carlos Vieira Silva; DURIGON, Edison Luiz; BOSCARDIN, Silvia Beatriz; ROSA, Daniela Santoro; SCHECHTMAN, Deborah; NAKAYA, Helder. I. I.; CUNHA-NETO, Edecio; GADERMAIER, Gabriele; KALIL, Jorge; COELHO, Veronica; SANTOS, Keity SouzaIntroductionConsidering the likely need for the development of novel effective vaccines adapted to emerging relevant CoV-2 variants, the increasing knowledge of epitope recognition profile among convalescents and afterwards vaccinated with identification of immunodominant regions may provide important information. MethodsWe used an RBD peptide microarray to identify IgG and IgA binding regions in serum of 71 COVID-19 convalescents and 18 vaccinated individuals. ResultsWe found a set of immunodominant RBD antibody epitopes, each recognized by more than 30% of the tested cohort, that differ among the two different groups and are within conserved regions among betacoronavirus. Of those, only one peptide, P44 (S415-429), recognized by 68% of convalescents, presented IgG and IgA antibody reactivity that positively correlated with nAb titers, suggesting that this is a relevant RBD region and a potential target of IgG/IgA neutralizing activity. DiscussionThis peptide is localized within the area of contact with ACE-2 and harbors the mutation hotspot site K417 present in gamma (K417T), beta (K417N), and omicron (K417N) variants of concern. The epitope profile of vaccinated individuals differed from convalescents, with a more diverse repertoire of immunodominant peptides, recognized by more than 30% of the cohort. Noteworthy, immunodominant regions of recognition by vaccinated coincide with mutation sites at Omicron BA.1, an important variant emerging after massive vaccination. Together, our data show that immune pressure induced by dominant antibody responses may favor hotspot mutation sites and the selection of variants capable of evading humoral response.
- Analysis of genetic polymorphisms associated with the presence of freckles for phenotypic prediction(2022) FRIDMAN, Cintia; FERREIRA, Marcelo Alves; MARANO, Leonardo Arduino; FORLENZA, Bettina StingelinThe prediction of externally visible characteristics (EVCs) is a commonly used practice by the forensic sciences as an important resource in the investigation of criminal cases in which the identity of perpetrators or victims is unknown or even to recognize decomposed cadavers. With this purpose, genetic markers associated with pigmentation traits have been widely studied by forensic scientists and, nowadays, it is possible to predict phenotypic characteristics such as hair, eyes and skin colour, as well as the presence of skin freckles by analysing single nucleotide polymorphisms (SNPs). In this study, we analysed the association of six SNPs located in pigmentation genes to the presence of freckles in individuals from the Brazilian population for forensic DNA phenotyping. The study was based within the context of a larger project on a population sample of 534 adult Brazilians of both sexes and different skin colours. DNA was extracted from peripheral blood and genotyped using the TaqMan & REG; OpenArray & REG; Real-Time PCR System (ThermoFischer Scientific) technique. Statistical analyses were carried out with the R software (version 4.0.2). As for the results obtained, three SNPs were shown to be statistically associated to the freckling, rs12203592, rs1800404 and rs222847, with CT, AG and AA genotypes being the main contributors, respectively. Variables such as sex of the individuals and skin colour were found to also contribute to the manifestation of this pigmentation trait. Further statistical analyses will be carried out to evaluate the possibility of using the SNPs in this study for phenotyping prediction of the Brazilian population, improving existing DNA phenotyping models in forensic sciences.
- Heparin Therapy Improving Hypoxia in COVID-19 Patients - A Case Series(2020) NEGRI, Elnara Marcia; PILOTO, Bruna Mamprim; MORINAGA, Luciana Kato; JARDIM, Carlos Viana Poyares; LAMY, Shari Anne El-Dash; FERREIRA, Marcelo Alves; D'AMICO, Elbio Antonio; DEHEINZELIN, DanielIntroduction Elevated D-dimer is a predictor of severity and mortality in COVID-19 patients, and heparin use during in-hospital stay has been associated with decreased mortality. COVID-19 patient autopsies have revealed thrombi in the microvasculature, suggesting that hypercoagulability is a prominent feature of organ failure in these patients. Interestingly, in COVID-19, pulmonary compliance is preserved despite severe hypoxemia corroborating the hypothesis that perfusion mismatch may play a significant role in the development of respiratory failure. Methods We describe a series of 27 consecutive COVID-19 patients admitted to Sirio-Libanes Hospital in Sao Paulo-Brazil and treated with heparin in therapeutic doses tailored to clinical severity. Results PaO2/FiO2 ratio increased significantly over the 72 h following the start of anticoagulation, from 254(+/- 90) to 325(+/- 80), p = 0.013, and 92% of the patients were discharged home within a median time of 11 days. There were no bleeding complications or fatal events. Discussion Even though this uncontrolled case series does not offer absolute proof that micro thrombosis in the pulmonary circulation is the underlying mechanism of respiratory failure in COVID-19, patient's positive response to heparinization contributes to the understanding of the pathophysiological mechanism of the disease and provides valuable information for the treatment of these patients while we await the results of further prospective controlled studies.
- Development, validation and application of a questionnaire to qualify the indoor environmental exposure of patients with respiratory allergy(2023) SOUZA, Barbara de; FERREIRA, Marcelo Alves; KALIL, Jorge; GIAVINA-BIANCHI, Pedro; AGONDI, Rosana CamaraObjective Environmental control includes measures to prevent exposure to common aeroallergens in an individual's home. Questionnaires are part of the clinical practice of health assessment, and are also widely used in research. Our aim was to develop and validate a questionnaire to identify possible sources of aeroallergens present in the indoor environment. Methods This study describes the development, validation and application of a questionnaire. For content validation the Content Validation Index and Ordinal Cronbach's Alpha Index have been used; Polychoric Correlations for the agreement between judges; and an Exploratory Factor Analysis for the structure of the questionnaire, while for reliability assessment, Intraclass Correlation Coefficient has been applied. Results Twenty-one doctors participated as judges to validate the questionnaire, which 204 patients answered. The Content Validity Index for all the questions on the ""Clarity"" aspect was 0.846 +/- 0.152 and on the ""Relevance"" aspect, 0.954 +/- 0.080. Cronbach's alpha coefficient for the ""Clarity"" aspect was 0.88 with a 95% confidence intervals (CI) and the ""Relevance"" aspect, 0.94 with a 95% CI. The average Intraclass Correlation Coefficient was 0.94 and all the F tests were highly significant. Conclusions The questionnaire developed by our group was considered valid and reliable, and is capable of portraying the home environment without the need for a personal visit to the patient's home. This questionnaire would be a good tool to use in research or during patient consultations to assess the patient's home environment, as this latter assessment is essential for the management of patients with respiratory allergies.
- The eggshell features and clutch viability of the broad-snouted caiman (Caiman latirostris) are associated with the egg burden of organochlorine compounds(2013) STOKER, C.; ZAYAS, M. A.; FERREIRA, M. A.; DURANDO, M.; GALOPPO, G. H.; RODRIGUEZ, H. A.; REPETTI, M. R.; BELDOMENICO, H. R.; CALDINI, E. G.; LUQUE, E. H.; MUNOZ-DE-TORO, M.Organochlorine compounds (OCCs) are toxic and have been identified as endocrine-disrupting chemicals (EDCs). The broad-snouted Caiman (Caiman latirostris) is an oviparous species widely distributed in South America with potential to accumulate OCCs. The eggshell is formed during passage of the eggs through the oviduct. Since the oviduct is a target of hormone actions, exposure to OCCs could modify eggshell quality, thus affecting clutch viability. Eight clutches were collected from wetlands of Parana River tributaries, in north-eastern Argentina. Two to four eggs per clutch were used to establish the burden of OCCs, eggshell thickness and eggshell porosity. The remaining eggs were incubated in controlled conditions. Ten days after hatching, hatchling survival was assessed. Organochlorine pesticide residues (OCP5) were found in all clutches, while polychlorinated biphenyls (PCBs) were present in all but one clutch. The principal contributors to the OCP burden were members of the DDT family and oxychlordane. Eggshell thickness was 400.9 +/- 6.0 mu m and, unexpectedly, no association between eggshell thickness and the OCC burden was found. The number of pores in the outer surface was 25.3 +/- 43 pores/cm(2). A significant inverse correlation between porosity and OCC burden was found (Pearson r = -0.81, p= 0.01). Furthermore, a decrease in caiman survival with decreased pore density was observed (Pearson r= 0.73, p = 0.04). Our findings highlight another potential negative impact of current and past use of OCCs on wildlife species.
- Neutrophils LL-37 migrate to the nucleus during overwhelming infection(2013) SILVA, Fabiano Pinheiro da; MEDEIROS, Maria Cristina Rodrigues; SANTOS, Angela Batista Gomes dos; FERREIRA, Marcelo Alves; GARIPPO, Ana Lucia; CHAMMAS, Roger; CALDINI, Elia; VELASCO, Irineu Tadeu; SOUZA, Heraldo Possolo de; MACHADO, Marcel Cerqueira CesarLL-37 is the only cathelicidin produced by human cells. It is secreted by a variety of cell types, including monocyte/macrophages, neutrophils, mast cells, keratinocytes and epithelial cells, acting on the extracellular milieu by directly killing bacteria or boosting innate immunity. Here, we show that LL-37 translocates to the nucleus following overwhelming infection, putting in evidence that its role may be even broader, with new potential important implications to cell biology. Future studies are necessary to address if LL-37 is able to induce or affect transcription, since it can lead to a novel cell signaling pathway that probably will contribute to the understanding of complex diseases.
- Reduced T Cell and Antibody Responses to Inactivated Coronavirus Vaccine Among Individuals Above 55 Years Old(2022) MEDEIROS, Giuliana X.; SASAHARA, Greyce Luri; MAGAWA, Jhosiene Y.; NUNES, Joao Paulo S.; BRUNO, Fernanda R.; KURAMOTO, Andreia C.; ALMEIDA, Rafael R.; FERREIRA, Marcelo A.; SCAGION, Guilherme P.; CANDIDO, Erika D.; LEAL, Fabyano B.; OLIVEIRA, Danielle B. L.; DURIGON, Edison L.; SILVA JR., Roberto Carlos V.; ROSA, Daniela S.; BOSCARDIN, Silvia B.; COELHO, Veronica; KALIL, Jorge; SANTOS, Keity S.; CUNHA-NETO, EdecioCoronaVac is an inactivated SARS-CoV-2 vaccine that has been rolled out in several low and middle-income countries including Brazil, where it was the mainstay of the first wave of immunization of healthcare workers and the elderly population. We aimed to assess the T cell and antibody responses of vaccinated individuals as compared to convalescent patients. We detected IgG against SARS-CoV-2 antigens, neutralizing antibodies against the reference Wuhan SARS-CoV-2 strain and used SARS-CoV-2 peptides to detect IFN-g and IL-2 specific T cell responses in a group of CoronaVac vaccinated individuals (N = 101) and convalescent (N = 72) individuals. The frequency among vaccinated individuals, of whom 96% displayed T cell and/or antibody responses to SARS-CoV-2, is comparable to 98.5% responses of convalescent individuals. We observed that among vaccinated individuals, men and individuals 55 years or older developed significantly lower anti-RBD, anti-NP and neutralization titers against the Wuhan strain and antigen-induced IL-2 production by T cells. Neutralizing antibody responses for Gamma variant were even lower than for the Wuhan strain. Even though some studies indicated CoronaVac helped reduce mortality among elderly people, considering the appearance of novel variants of concern, CoronaVac vaccinated individuals above 55 years old are likely to benefit from a heterologous third dose/booster vaccine to increase immune response and likely protection.