ItemA taxonomic review of the genus Paracoccidioides, with focus on the uncultivable species(2023-04-01) Vilela, Raquel; de Hoog, Sybren; Bensch, Konstanze; Bagagli, Eduardo [UNESP]; Mendoza, Leonel; Universidade Federal de Minas Gerais (UFMG); Michigan State University; Center of Expertise in Mycology of RadboudUMC/Canisius Wilhelmina Hospital; Westerdijk Fungal Biodiversity Institute; Universidade Estadual Paulista (UNESP)Paracoccidioides species have always been surrounded by taxonomic uncertainties. The continuing nomenclatoral muddle was caused in part by the failure of Adolfo Lutz and Jorge Lôbo to name the etiologic agents of human paracoccidioidomycosis and Jorge Lôbo’s diseases, respectively. Early in their history, it was postulated that the cultivable species causing systemic infections belonged in the genus Paracoccidioides, whereas the uncultivable species, causing skin disease, were not part of the genus. The taxonomy of these pathogens was further complicated when a similar skin disease with numerous yeast-like cells in infected dolphins was also reported. Due to its phenotypic similarities with that described by Jorge Lôbo in human and its uncultivable nature, it was assumed that the disease in dolphins was caused by the same fungus. Recent molecular and population genetic analysis, however, found the DNA extracted from the uncultivable yeast-like cells affecting dolphins shared common phylogenetic traits with cultivable Paracoccidioides species. The study revealed that the uncultivable pathogens comprised 2 different Paracoccidioides species, now known as P. ceti and P. loboi, correspondingly. To validate P. loboi binomial, a comprehensive historical critical review of Jorge Lôbo etiology was performed. This review showed the proposed binomial P. loboi was previously used, and, thus, a replacement name is introduced, Paracoccidioides lobogeorgii nom. nov. In addition, in this review, several cultivable human Paracoccidioides species are validated, and the generic type species, P. brasiliensis, is neotypified as the original material could not be traced.. ItemIntegrative Analysis of the Ethanol Tolerance of Saccharomyces cerevisiae(2023-03-01) Wolf, Ivan Rodrigo [UNESP]; Marques, Lucas Farinazzo [UNESP]; de Almeida, Lauana Fogaça [UNESP]; Lázari, Lucas Cardoso [UNESP]; de Moraes, Leonardo Nazário [UNESP]; Cardoso, Luiz Henrique [UNESP]; Alves, Camila Cristina de Oliveira [UNESP]; Nakajima, Rafael Takahiro [UNESP]; Schnepper, Amanda Piveta [UNESP]; Golim, Marjorie de Assis [UNESP]; Cataldi, Thais Regiani; Nijland, Jeroen G.; Pinto, Camila Moreira [UNESP]; Fioretto, Matheus Naia [UNESP]; Almeida, Rodrigo Oliveira; Driessen, Arnold J. M.; Simōes, Rafael Plana [UNESP]; Labate, Mônica Veneziano; Grotto, Rejane Maria Tommasini [UNESP]; Labate, Carlos Alberto; Fernandes Junior, Ary [UNESP]; Justulin, Luis Antonio [UNESP]; Coan, Rafael Luiz Buogo [UNESP]; Ramos, Érica [UNESP]; Furtado, Fabiana Barcelos [UNESP]; Martins, Cesar [UNESP]; Valente, Guilherme Targino; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); University of Groningen; Ciência e Tecnologia do Sudeste de Minas Gerais; Max Planck Institute for Heart and Lung ResearchEthanol (EtOH) alters many cellular processes in yeast. An integrated view of different EtOH-tolerant phenotypes and their long noncoding RNAs (lncRNAs) is not yet available. Here, large-scale data integration showed the core EtOH-responsive pathways, lncRNAs, and triggers of higher (HT) and lower (LT) EtOH-tolerant phenotypes. LncRNAs act in a strain-specific manner in the EtOH stress response. Network and omics analyses revealed that cells prepare for stress relief by favoring activation of life-essential systems. Therefore, longevity, peroxisomal, energy, lipid, and RNA/protein metabolisms are the core processes that drive EtOH tolerance. By integrating omics, network analysis, and several other experiments, we showed how the HT and LT phenotypes may arise: (1) the divergence occurs after cell signaling reaches the longevity and peroxisomal pathways, with CTA1 and ROS playing key roles; (2) signals reaching essential ribosomal and RNA pathways via SUI2 enhance the divergence; (3) specific lipid metabolism pathways also act on phenotype-specific profiles; (4) HTs take greater advantage of degradation and membraneless structures to cope with EtOH stress; and (5) our EtOH stress-buffering model suggests that diauxic shift drives EtOH buffering through an energy burst, mainly in HTs. Finally, critical genes, pathways, and the first models including lncRNAs to describe nuances of EtOH tolerance are reported here. ItemPropolis consumption by asymptomatic HIV-individuals: Better redox state? A prospective, randomized, double-blind, placebo-controlled trial(2023-06-01) Tasca, Karen Ingrid [UNESP]; Conte, Fernanda Lopes [UNESP]; Correa, Camila Renata [UNESP]; Santiago, Karina Basso [UNESP]; Cardoso, Eliza de Oliveira [UNESP]; Manfio, Vanessa Martinez [UNESP]; Garcia, Jessica Leite [UNESP]; Berretta, Andresa Aparecida; Sartori, Arthur Alves [UNESP]; Honorio, Mariana da Silva [UNESP]; Souza, Lenice do Rosário [UNESP]; Sforcin, José Maurício [UNESP]; Universidade Estadual Paulista (UNESP); R&D DepartmentPropolis is a natural product has many biological properties of clinical interest, such as anti-inflammatory and antioxidant. Considering that people living with HIV/aids (PLWHA) on effective combined antiretroviral therapy (cART) present early aging due to an intense immune activation, inflammation, and redox imbalance, propolis consumption could offer a benefit to such patients. This double-blind longitudinal study evaluated whether Brazilian green propolis pills intake (500 mg/day for three months) would decrease the oxidative stress of virological suppressed HIV-individuals. To compare each group (propolis, n = 20 versus placebo, n = 20) in both moments (M0, before and M1, after the intervention), the following markers were assessed: plasma malondialdehyde (MDA), carbonylation, total oxide nitric, total antioxidant capacity (TAP), superoxide dismutase, catalase, and NFkB and NRF2 gene expression. Data were analyzed using Poisson, Gamma distribution and ANOVA followed by Tukey-Kramer. The groups were homogeneous regarding age, gender, time of diagnosis/ treatment, cART scheme, CD4+ T cell count, and no changes were observed in the diet food, or patients’ lifestyles. A decreased MDA concentration was seen in the propolis group (M0 = 0.24 ± 0.13, M1 = 0.20 ± 0.10 protein nmol/mg; p = 0.005) as well as a slight but non-significant increase of TAP (M0 = 49.07 ± 13.26, M1 = 52.27 ± 14.86%; p = 0.06). One may conclude that propolis promoted a lower lipid peroxidation and improved the antioxidant system, suggesting that its use may be beneficial to PLWHA in an attempt to contain the intense inflammatory and oxidant activity. ItemRelationship between Biofilm Production and High Somatic Cell Count in Streptococcus agalactiae Isolated from Milk of Cows with Subclinical Mastitis(2023-02-01) Bonsaglia, Erika Carolina Romão [UNESP]; Rossi, Rodolfo S. [UNESP]; Latosinski, Giulia [UNESP]; Rossi, Bruna Fernanda [UNESP]; Campos, Fernanda Cristina [UNESP]; Junior, Ary Fernandes [UNESP]; Pantoja, José Carlos F. [UNESP]; Rall, Vera Lucia Mores [UNESP]; Universidade Estadual Paulista (UNESP)Streptococcus agalactiae (S. agalactiae) is one of the main agents that causes mastitis in dairy cows, mainly inducing the subclinical form, which is characterized by a high somatic cell count (SCC). The aim of this study was to correlate the increase in SCC caused by S. agalactiae in cows with subclinical mastitis to the presence of genes related to adhesion and invasion in bovine mammary epithelial cells (BMEC) and biofilm formation. Considering the 145 isolates tested, 57.2% presented the capsular type Ia and 42.8% presented type III. We identified the virulence genes among the isolates and determined nine genetic profiles. The most common profile was identified in 69 isolates (47.5%): Ia, fbsA+, fbsB-, pI1-, pI2a-, pI2b+, and hylb+. All isolates produced biofilm, with 58.6% classified as strong producers, 29% as moderate producers and 12.4% as weak producers. No statistical correlation was found between the presence of virulence genes and increased SCC or biofilm production. However, biological evidence was observed between increased SCC and biofilm production. One isolate from each profile was randomly subjected to adhesion and invasion assays, and all of them adhered to BEMC, but none were able to invade. Our results showed that different genetic profiles do not provide advantages for bacteria to invade BMEC in vitro. In addition, biofilm production appears to be related to high SCC. ItemExosomes: Pros and cons for fighting cancer(2020-09-24) Romagnoli, Graziela Gorete [UNESP]; Gorgulho, Carolina Mendonça [UNESP]; Kaneno, Ramon [UNESP]; Universidade Estadual Paulista (UNESP); Oeste Paulista University - UNOESTEExosomes (Exo) are extracellular vesicles involved in intercellular communication. These nanovesicles are a mini reflection of the cell of origin, carrying several bioactive molecules, also reflecting the conditions of the extracellular environment. Exo can be a double-edged sword depending on their origin, so that tumor cell-derived Exo may be loaded with important tumor antigens to be presented to the immune system, but may also decrease the immunotherapeutic action of monoclonal antibodies. Moreover, tumor Exo carry many molecules with suppressive potential, subverting the immune response. Alternatively, Exo originating from antigen-presenting cells (APCs) such as dendritic cells (DCs) may directly or indirectly induce tumor-specific T lymphocyte response, both in vitro and in vivo, being clinically safe. Another clinically attractive factor of Exo is its use as a noninvasive liquid biopsy, which may facilitate diagnosis in the early phase of the disease. In addition, they can be used as biotechnological tools for loading bioactive drugs. ItemBrazilian red propolis in combination with β-lactams exerts an efficient antibacterial action over methicillin-resistant Staphylococcus aureus (MRSA) strains(2023-02-16) Ripari, Nicolas [UNESP]; Pereira, Ana Flávia Marques [UNESP]; Júnior, Ary Fernandes [UNESP]; Rall, Vera Lúcia Mores [UNESP]; Aldana-Mejía, Jennyfer A.; Bastos, Jairo Kenupp; Sforcin, José Maurício [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP)AIMS: The antibacterial activity of red propolis extract (RPE) and brown propolis extracts (BPE) and the synergistic effect of RPE with cefoxitin (CEFO), imipenem (IMI), and ertapenem (ERTA) was evaluated in vitro against methicillin-resistant Staphylococcus aureus (MRSA) strains. METHODS AND RESULTS: MRSA ATCC 33591, community-associated (CA-MRSA) USA300, and four clinical isolates were used. A broth microdilution assay was performed to obtain inhibitory and bactericidal concentrations of BPE, RPE, CEFO, IMI, and ERTA. RPE in combination with CEFO, IMI, and ERTA was evaluated on the formation or eradication of biofilm. The bacterial relative membrane conductivity of the strains was assessed after RPE and combinations exposition. Surface/binding computational analyzes between RPE compounds and penicillin binding protein 2a (PBP2a) were performed. BPE samples had no activity against MRSA (MICs 3.2-5 g l-1; MBCs 10-15 g l-1), so the subsequent assays were carried out only with RPE and antimicrobials. RPE exerted a bacteriostatic action (MICs 0.0156-0.125 g l-1; MBCs 0.5-2 g l-1) but the combinations with IMI and ERTA showed the highest inhibition, as observed in the time-kill curve. However, the FICI index showed synergism (≥0.5) only to RPE + IMI. This combination was the most effective in inhibiting the biofilm and showed the highest values of membrane conductivity. Computational predictions indicated that RPE constituents may interact with PBP2a. CONCLUSION: RPE and RPE + IMI exerted an antibacterial and antibiofilm activity on MRSA strains probably due to membrane/wall damage and interactions with PBP2a. ItemInvestigation of the Presence of Leptospira interrogans in Urinary and Genital Tracts of Male Goats Raised in the Semiarid Region of Brazil(2023-01-01) Soares, Rafael Rodrigues; da Costa Barnabé, Nathanael Natércio; Júnior, João Pessoa Araújo [UNESP]; Malossi, Camila Dantas [UNESP]; Ullmann, Leila Sabrina [UNESP]; da Costa, Diego Figueiredo; Rodrigues Silva, Maria Luana Cristiny; dos Santos Higino, Severino Silvano; de Azevedo, Sergio Santos; Alves, Clebert José; Av. Universitária; Universidade Estadual Paulista (UNESP); Universidade Federal da Paraíba (UFPB)Leptospira sp. is a bacterium present in different environments, including those with unfavorable conditions for survival, with evidence pointing to its adaptation to the genital tract. The aim of the study was to detect Leptospira sp. using serological, molecular and bacterial techniques in samples from the genital and urinary tracts of male goats slaughtered under semiarid conditions. Forty animals destined for slaughter were sampled and, from each one, samples were taken from the urinary and genital tracts for analysis by polymerase chain reaction (PCR) and microbial culture growth, as well as serum samples for microscopic agglutination testing (MAT). Serological data showed three (7.5%) animals positive for the Pyrogenes serogroup. DNA from Leptospira sp. was detected by molecular testing in 25/120 (20.8%) of the urinary tract samples and in 23/120 (19.1%) of the genital tracts, with no statistically significant difference. A Leptospira sp. sample collected from the vas deferens was sent for DNA sequencing and showed 99% similarity with L. interrogans. Overall, 240 cultures were evaluated and, of these, 36 (15%) showed bacterial growth. Thus, Leptospira sp. organisms were present in male goats managed in an environment of adverse conditions and the spread of Leptospira sp. may be linked to venereal transmission. ItemInhibitory activities of propolis, nisin, melittin and essential oil compounds on Paenibacillus alvei and Bacillus subtilis(2022-01-01) Sani, Alessandra Aguirra [UNESP]; Pereira, Ana Flávia Marques [UNESP]; Furlanetto, Alessandra [UNESP]; de Sousa, Débora Silva Marques [UNESP]; Zapata, Tatiane Baptista [UNESP]; Rall, Vera Lucia Mores [UNESP]; Júnior, Ary Fernandes [UNESP]; Universidade Estadual Paulista (UNESP)Background: Natural products represent important sources of antimicrobial compounds. Propolis and compounds from essential oils comprise good examples of such substances because of their inhibitory effects on bacterial spores, including bee pathogens. Methods: Ethanol extracts of propolis (EEP) from Apis mellifera were prepared using different methods: double ultrasonication, double maceration and maceration associated with ultrasonication. Together with the antimicrobial peptides nisin and melittin, and compounds present in the essential oils of clove (Syzygium aromaticum) and cinnamon (Cinnamomum zeylanicum), assays were carried out on one Bacillus subtilis isolate and Paenibacillus alvei (ATCC 6344) against vegetative and sporulated forms, using the resazurin microtiter assay. Synergism with all the antimicrobials in association with tetracycline was verified by the time-kill curve method. Potassium and phosphate efflux, release of proteins and nucleic acids were investigated. Results: EEPs showed the same MIC, 156.25 µg/mL against B. subtilis and 78.12 µg/mL against P. alvei. The peptides showed better activities against B. subtilis (MIC of 12 µg/ mL for melittin and 37.50 µg/mL for nisin). Antimicrobials showed similar inhibitory effects, but cinnamaldehyde (39.06 µg/mL) showed the best action against P. alvei. Melittin and nisin showed the greatest capacity to reduce spores, regarding B. subtilis there was a 100% reduction at 6.25 and 0.78 µg/mL, respectively. Concerning P. alvei, the reduction was 93 and 98% at concentrations of 80 µg/mL of melittin and 15 µg/ mL of nisin. EEPs showed the highest effects on the protein release against B. subtilis and P. alvei. Nucleic acid release, phosphate and potassium efflux assays indicated bacterial cell membrane damage. Synergism between antimicrobials and tetracycline was demonstrated against both bacteria. Conclusion: All antimicrobials tested showed antibacterial activities against vegetative and sporulated forms of P. alvei and B. subtilis, especially nisin and melittin. Synergism with tetracycline and damage on bacterial cell membrane also occurred. ItemMolecular detection of Trypanosoma cruzi in equids in a semiarid zone of Pernambuco - Brazil(2023-01-01) Limeira, Clécio Henrique; Oliveira, Murilo Duarte; Júnior, João Pessoa Araújo [UNESP]; Malossi, Camila Dantas [UNESP]; Ullmann, Leila Sabrina [UNESP]; Silva, Maria Luana Cristiny Rodrigues; Azevedo, Sérgio Santos; Alves, Clebert José; Universidade Federal de Campina Grande; Instituto Federal de Educação do Sertão Pernambucano; Universidade Estadual Paulista (UNESP); Universidade Federal de Mato Grosso do Sul (UFMS)Trypanosoma cruzi is the etiological agent of Chagas disease, a neglected and frequently occurring zoonosis in Central and South American countries. Wild mammals and domestic dogs are the main reservoirs of the parasite in the wild and domestic cycles, respectively. The vectors have a wide variety of food sources that can influence transmission cycles. The aim of this study was to determine the prevalence of T. cruzi infection in donkeys (Equus asininos) and mules (Equus mulus) living in rural areas of the Brazilian semi-arid region. Whole-blood samples from 72 equids (65 donkeys and 7 mules) were analyzed by nested polymerase chain reaction (nested PCR). A total of 51.39% of the samples (37/72) were positive. Phylogenetic analysis identified discrete typing units TcI and TcII, which suggested the possibility that donkeys and mules might be participating in domestic/peridomestic and wild transmission cycles. This was the first report of T. cruzi infection in donkeys and mules in Brazil, with high prevalence of positive animals. This places these animals as potential reservoirs for the parasite and the particular features of these hosts, the presence of vectors and the socioeconomic characteristics of the population under semiarid conditions create interactions that may favor transmission and overlapping T. cruzi infection cycles. ItemMicroarray analysis of canine osteosarcoma cells exposed to Colombian propolis(2022-11-01) Pardo-Mora, Dolly Patricia; Murillo, Oscar Julián; Buitrago, Mauricio Rey; Rodríguez, Anny; Uribe, Jaime Fabian Cruz; Sforcin, José Maurício [UNESP]; García, Orlando Torres; Antonio Nariño University; Universidad Nacional de Colombia; Universidade Estadual Paulista (UNESP)Background: Propolis is a bee product exhibiting a cytotoxic activity against tumor cells. This study aimed at investigating the gene expression profile of canine osteosarcoma cells exposed to Colombian propolis by microarray analysis, in an attempt to identify differentially expressed genes and the pathways involved in propolis cytotoxic action. Methods: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase assays were used to assess the viability and the half-maximal inhibitory concentration for OSCA-8 cells and fibroblasts after incubation with propolis samples, its solvent (70% ethanol) or doxorubicin (positive control). The gene expression profile was determined by the Canine Gene 1.0 ST Array (Affymetrix) and the microarray data was validated by real time PCR. Enrichment and annotation analysis were applied for the differentially expressed genes. Results: Propolis reduced cell viability and exerted a cytotoxic effect on OSCA-8 cells, exhibiting a selective effect in relation to non-tumor cells. The microarray analysis showed changes in more than 253 genes involved in intracellular pathways related to the inflammatory response, growth factor activity and chemotaxis, which are associated with cancer development. Propolis exerted doxorubicin-like changes in gene expression. Conclusion: Propolis and Dox exerted a similar action, altering some pathways and indicating possible mechanisms of action involved in the cytotoxic effect on OSCA-8 cells. ItemSem título(2023-01-01) de Castro, Mateus V.; Silva, Monize V. R.; Soares, Flávia B.; Cória, Vivian R.; Naslavsky, Michel S.; Scliar, Marilia O.; Castelli, Erick C. [UNESP]; de Oliveira, Jamile R.; de Medeiros, Giuliana X.; Sasahara, Greyce L.; Santos, Keity S.; Cunha-Neto, Edecio; Kalil, Jorge; Zatz, Mayana; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); Instituto Nacional de Ciências e Tecnologia-iii-INCTIn the published article, there was an error in the author list, and author Giuliana X. de Medeiros was erroneously excluded. The corrected author list appears below. Mateus V. de Castro1†, Monize V. R. Silva1†, Flávia B. Soares1, 2†, Vivian R. Cória1, Michel S. Naslavsky1, 2, Marilia O. Scliar1, 2, Erick C. Castelli3, Jamile R. de Oliveira4, 5, Giuliana X. de Medeiros4, 5, Greyce L. Sasahara5, Keity S. Santos4, 5, 6, Edecio Cunha-Neto4, 5, 6, Jorge Kalil4, 5, 6 and Mayana Zatz1, 2* The authors apologize for this error and state that this does not change the scientific conclusions of the article in any way. The original article has been updated. ItemA novel insight on SARS-CoV-2 S-derived fragments in the control of the host immunity(2023-12-01) Bastos, Thais Sibioni Berti; de Paula, André Guilherme Portela; dos Santos Luz, Rebeca Bosso; Garnique, Anali M. B.; Belo, Marco A. A.; Eto, Silas Fernandes; Fernandes, Dayanne Carla; Ferraris, Fausto Klabund; de Pontes, Leticia Gomes; França, Tábata Takahashi; Barcellos, Leonardo José Gil; Veras, Flavio P.; Bermejo, Pamela; Guidelli, Giovanna; Maneira, Carla; da Silveira Bezerra de Mello, Fellipe; Teixeira, Gleidson; Pereira, Gonçalo Amarante Guimarães; Fernandes, Bianca H. Ventura; Sanches, Paulo R. S. [UNESP]; Braz, Helyson Lucas Bezerra; Jorge, Roberta Jeane Bezerra; Malafaia, Guilherme; Cilli, Eduardo M. [UNESP]; Olivier, Danilo da Silva; do Amaral, Marcos Serrou; Medeiros, Renata J.; Condino-Neto, Antonio; Carvalho, Luciani R.; Machado-Santelli, Glaucia M.; Charlie-Silva, Ives; Galindo-Villegas, Jorge; Braga, Tárcio Teodoro; Federal University of Parana; Instituto Carlos Chagas; Universidade de São Paulo (USP); Brasil University; Butantan Institute; Veterinarian; FIOCRUZ; University São Paulo; University of Passo Fundo; Federal University of Santa Maria; Universidade Estadual de Campinas (UNICAMP); Nord University; Universidade Estadual Paulista (UNESP); Federal University of Ceará; Urutai Campus; Federal University of Tocantins; Federal University of Mato Grosso do Sul; National Institute for Quality Control in HealthDespite all efforts to combat the pandemic of COVID-19, we are still living with high numbers of infected persons, an overburdened health care system, and the lack of an effective and definitive treatment. Understanding the pathophysiology of the disease is crucial for the development of new technologies and therapies for the best clinical management of patients. Since the manipulation of the whole virus requires a structure with an adequate level of biosafety, the development of alternative technologies, such as the synthesis of peptides from viral proteins, is a possible solution to circumvent this problem. In addition, the use and validation of animal models is of extreme importance to screen new drugs and to compress the organism's response to the disease. Peptides derived from recombinant S protein from SARS-CoV-2 were synthesized and validated by in silico, in vitro and in vivo methodologies. Macrophages and neutrophils were challenged with the peptides and the production of inflammatory mediators and activation profile were evaluated. These peptides were also inoculated into the swim bladder of transgenic zebrafish larvae at 6 days post fertilization (dpf) to mimic the inflammatory process triggered by the virus, which was evaluated by confocal microscopy. In addition, toxicity and oxidative stress assays were also developed. In silico and molecular dynamics assays revealed that the peptides bind to the ACE2 receptor stably and interact with receptors and adhesion molecules, such as MHC and TCR, from humans and zebrafish. Macrophages stimulated with one of the peptides showed increased production of NO, TNF-α and CXCL2. Inoculation of the peptides in zebrafish larvae triggered an inflammatory process marked by macrophage recruitment and increased mortality, as well as histopathological changes, similarly to what is observed in individuals with COVID-19. The use of peptides is a valuable alternative for the study of host immune response in the context of COVID-19. The use of zebrafish as an animal model also proved to be appropriate and effective in evaluating the inflammatory process, comparable to humans. ItemJoint Association between Sedentary Time and Moderate-to-Vigorous Physical Activity with Obesity Risk in Adults from Latin America(2023-04-01) de Victo, Eduardo Rossato; Fisberg, Mauro; Solé, Dirceu; Kovalskys, Irina; Gómez, Georgina; Rigotti, Attilio; Cortes, Lilia Yadira; Yépez-Garcia, Martha Cecilia; Pareja, Rossina; Herrera-Cuenca, Marianella; Drenowatz, Clemens; Christofaro, Diego [UNESP]; Araujo, Timoteo; Silva, Danilo; Ferrari, Gerson; Universidade Federal de São Paulo (UNIFESP); Hospital Infantil Sabará; Pontificia Universidad Católica Argentina; Universidad de Costa Rica; Pontificia Universidad Católica; Pontificia Universidad Javeriana; Universidad San Francisco de Quito; Instituto de Investigación Nutricional; Universidad Central de Venezuela (CENDES-UCV)/Fundación Bengoa; University of Education Upper Austria; Universidade Estadual Paulista (UNESP); Faculdades Metropolitanas Unidas; Universidade Federal de Sergipe (UFS); Universidad Autónoma de ChileRecent studies have shown various relationships between physical activity and the incidence of obesity, but this study critically explored the association of sedentary time (ST) and moderate-to-vigorous physical activity (MVPA) with obesity risk in adults from eight Latin American countries. ST and MVPA were assessed with accelerometers and stratified into 16 joint categories. Multivariate logistic regression models were used. The obesity risk indicators evaluated were body mass index (BMI), waist circumference (WC), and neck circumference (NC). Quartile 4 of ST and ≥300 min/week of MVPA was associated with lower odds of BMI compared to quartile 1 of ST and ≥300 min/week of MVPA. Quartile 1 of ST and 150–299 min/week of MVPA, quartile 1 of ST and 76–149 min/week MVPA, quartile 3 of ST and 76–149 min/week MVPA, and quartiles 1, 2, and 3 of ST with 0–74 min/week MVPA were associated with higher odds of high WC compared to quartile 1 of ST and ≥300 min/week of MVPA. Quartile 3 of ST and 150–299 min/week of MVPA, quartiles 1 and 3 of ST and 76–149 min/week of MVPA, and quartile 1 of ST and 0–74 min/week MVPA were associated with higher NC compared to quartile 1 of ST and ≥300 min/week of MVPA. This study suggests that achieving the MVPA recommendations will likely protect against obesity, regardless of ST. ItemLung Inflammation Induced by Inactivated SARS-CoV-2 in C57BL/6 Female Mice Is Controlled by Intranasal Instillation of Vitamin D(2023-04-01) Fernandes de Souza, William Danilo [UNESP]; Zorzella-Pezavento, Sofia Fernanda Gonçalves [UNESP]; Ayupe, Marina Caçador; Salgado, Caio Loureiro; Oliveira, Bernardo de Castro; Moreira, Francielly; da Silva, Guilherme William; Muraro, Stefanie Primon; de Souza, Gabriela Fabiano; Proença-Módena, José Luiz; Araujo Junior, Joao Pessoa [UNESP]; Fonseca, Denise Morais da; Sartori, Alexandrina [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP); Universidade Estadual de Campinas (UNICAMP)The COVID-19 pandemic was triggered by the coronavirus SARS-CoV-2, whose peak occurred in the years 2020 and 2021. The main target of this virus is the lung, and the infection is associated with an accentuated inflammatory process involving mainly the innate arm of the immune system. Here, we described the induction of a pulmonary inflammatory process triggered by the intranasal (IN) instillation of UV-inactivated SARS-CoV-2 in C57BL/6 female mice, and then the evaluation of the ability of vitamin D (VitD) to control this process. The assays used to estimate the severity of lung involvement included the total and differential number of cells in the bronchoalveolar lavage fluid (BALF), histopathological analysis, quantification of T cell subsets, and inflammatory mediators by RT-PCR, cytokine quantification in lung homogenates, and flow cytometric analysis of cells recovered from lung parenchyma. The IN instillation of inactivated SARS-CoV-2 triggered a pulmonary inflammatory process, consisting of various cell types and mediators, resembling the typical inflammation found in transgenic mice infected with SARS-CoV-2. This inflammatory process was significantly decreased by the IN delivery of VitD, but not by its IP administration, suggesting that this hormone could have a therapeutic potential in COVID-19 if locally applied. To our knowledge, the local delivery of VitD to downmodulate lung inflammation in COVID-19 is an original proposition. ItemCOVID-19 and Multiple Sclerosis: A Complex Relationship Possibly Aggravated by Low Vitamin D Levels(2023-03-01) Fernandes de Souza, William Danilo [UNESP]; Fonseca, Denise Morais da; Sartori, Alexandrina [UNESP]; Universidade Estadual Paulista (UNESP); Universidade de São Paulo (USP)Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an exceptionally transmissible and pathogenic coronavirus that appeared at the end of 2019 and triggered a pandemic of acute respiratory disease, known as coronavirus disease 2019 (COVID-19). COVID-19 can evolve into a severe disease associated with immediate and delayed sequelae in different organs, including the central nervous system (CNS). A topic that deserves attention in this context is the complex relationship between SARS-CoV-2 infection and multiple sclerosis (MS). Here, we initially described the clinical and immunopathogenic characteristics of these two illnesses, accentuating the fact that COVID-19 can, in defined patients, reach the CNS, the target tissue of the MS autoimmune process. The well-known contribution of viral agents such as the Epstein-Barr virus and the postulated participation of SARS-CoV-2 as a risk factor for the triggering or worsening of MS are then described. We emphasize the contribution of vitamin D in this scenario, considering its relevance in the susceptibility, severity and control of both pathologies. Finally, we discuss the experimental animal models that could be explored to better understand the complex interplay of these two diseases, including the possible use of vitamin D as an adjunct immunomodulator to treat them. ItemThe + 3010/C single nucleotide polymorphism (rs1710) at the HLA-G 3′ untranslated region is associated with a short transcript exhibiting a deletion of 92 nucleotides(2023-04-01) Castelli, Erick C. [UNESP]; Paes, Gabriela Sato [UNESP]; da Silva, Isabelle Mira [UNESP]; Moreau, Philippe; Donadi, Eduardo A.; Universidade Estadual Paulista (UNESP); Service de Recherches en Hémato-ImmunologieHôpital Saint-Louis; Université Paris Cité; Universidade de São Paulo (USP)The physiological expression of HLA-G is mainly observed in the placenta, playing an essential role in maternal–fetal tolerance. Among the HLA-G mRNA alternative transcripts, the one lacking 92 bases at the HLA-G 3′ untranslated region (3′UTR), the 92bDel transcript, is more stable, is associated with increased HLA-G soluble levels, and was observed in individuals presenting a 14 bp insertion (14 bp+) at the 3′UTR. We investigated the presence of the 92bDel transcript in placenta samples, correlating its expression levels with the HLA-G polymorphisms at the 3′UTR. The 14 bp+ allele correlates with the presence of the 92bDel transcript. However, the polymorphism triggering this alternative splicing is the + 3010/C allele (rs1710, allele C). Most 14 bp+ haplotypes (UTR-2/-5/-7) present allele + 3010/C. However, 14 bp− haplotypes such as UTR-3 are also associated with + 3010/C, and the 92bDel transcript can be detected in homozygous samples for the 14 bp- allele carrying at least one copy of UTR-3. The UTR-3 haplotype is associated with alleles G*01:04 and the HLA-G lineage HG0104, which is a high-expressing lineage. The only HLA-G lineage that is not likely to produce this transcript is HG010101, associated with the + 3010/G allele. This functional difference may be advantageous, considering the high worldwide frequency of the HG010101 lineage. Therefore, HLA-G lineages are functionally distinct regarding the 92bDel transcript expression, and the 3010/C allele triggers the alternative splicing that produces this shorter and more stable transcript. ItemImmunological evaluation of young unvaccinated patients with Turner syndrome after COVID-19(2023-04-01) de Castro, Mateus V.; Silva, Monize V.R.; Oliveira, Luana de M.; Gozzi-Silva, Sarah C.; Naslavsky, Michel S.; Scliar, Marilia O.; Magalhães, Monize L.; da Rocha, Katia M.; Nunes, Kelly; Castelli, Erick C. [UNESP]; Magawa, Jhosiene Y.; Santos, Keity S.; Cunha-Neto, Edecio; Sato, Maria N.; Zatz, Mayana; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); Instituto de Investigação em Imunologia - Instituto Nacional de Ciências e Tecnologia-iii-INCTObjectives: The X-chromosome contains the largest number of immune-related genes, which play a major role in COVID-19 symptomatology and susceptibility. Here, we had a unique opportunity to investigate, for the first time, COVID-19 outcomes in six unvaccinated young Brazilian patients with Turner syndrome (TS; 45, X0), including one case of critical illness in a child aged 10 years, to evaluate their immune response according to their genetic profile. Methods: A serological analysis of humoral immune response against SARS-CoV-2, phenotypic characterization of antiviral responses in peripheral blood mononuclear cells after stimuli, and the production of cytotoxic cytokines of T lymphocytes and natural killer cells were performed in blood samples collected from the patients with TS during the convalescence period. Whole exome sequencing was also performed. Results: Our volunteers with TS showed a delayed or insufficient humoral immune response to SARS-CoV-2 (particularly immunoglobulin G) and a decrease in interferon-γ production by cluster of differentiation (CD)4+ and CD8+ T lymphocytes after stimulation with toll-like receptors 7/8 agonists. In contrast, we observed a higher cytotoxic activity in the volunteers with TS than the volunteers without TS after phorbol myristate acetate/ionomycin stimulation, particularly granzyme B and perforin by CD8+ and natural killer cells. Interestingly, two volunteers with TS carry rare genetic variants in genes that regulate type I and III interferon immunity. Conclusion: Following previous reports in the literature for other conditions, our data showed that patients with TS may have an impaired immune response against SARS-CoV-2. Furthermore, other medical conditions associated with TS could make them more vulnerable to COVID-19. ItemComputational and atomistic studies applied to the understanding of the structural and behavioral features of the immune checkpoint HLA-G molecule and gene(2023-01-01) Alves, Cinthia C.; Arns, Thaís; Oliveira, Maria L.; Moreau, Philippe; Antunes, Dinler A.; Castelli, Erick C. [UNESP]; Mendes-Junior, Celso T.; Giuliatti, Silvana; Donadi, Eduardo A.; Universidade de São Paulo (USP); Luxembourg Centre for Systems Biomedicine; Service de Recherches en Hémato-Immunologie; Université Paris-Cité; University of Houston; Universidade Estadual Paulista (UNESP)We took advantage of the increasingly evolving approaches for in silico studies concerning protein structures, protein molecular dynamics (MD), protein-protein and protein-DNA docking to evaluate: (i) the structure and MD characteristics of the HLA-G well-recognized isoforms, (ii) the impact of missense mutations at HLA-G receptor genes (LILRB1/2), and (iii) the differential binding of the hypoxia-inducible factor 1 (HIF1) to hypoxia-responsive elements (HRE) at the HLA-G gene. Besides reviewing these topics, they were revisited including the following novel results: (i) the HLA-G6 isoforms were unstable docked or not with β2-microglobulin or peptide, (ii) missense mutations at LILRB1/2 genes, exchanging amino acids at the intracellular domain, particularly those located within and around the ITIM motifs, may impact the HLA-G binding strength, and (iii) HREs motifs at the HLA-G promoter or exon 2 regions exhibiting a guanine at their third position present a higher affinity for HIF1 when compared to an adenine at the same position. These data shed some light into the functional aspects of HLA-G, particularly how polymorphisms may influence the role of the molecule. Computational and atomistic studies have provided alternative tools for experimental physical methodologies, which are time-consuming, expensive, demanding large quantities of purified proteins, and exhibit low output. ItemSem título(2022-12-01) de Castro, Mateus V.; Silva, Monize V. R.; Naslavsky, Michel S.; Scliar, Marilia O.; Nunes, Kelly; Passos-Bueno, Maria Rita; Castelli, Erick C. [UNESP]; Magawa, Jhosiene Y.; Adami, Flávia L.; Moretti, Ana I. S.; de Oliveira, Vivian L.; Boscardin, Silvia B.; Cunha-Neto, Edecio; Kalil, Jorge; Jouanguy, Emmanuelle; Bastard, Paul; Casanova, Jean-Laurent; Quiñones-Vega, Mauricio; Sosa-Acosta, Patricia; Guedes, Jéssica de S.; de Almeida, Natália P.; Nogueira, Fábio C. S.; Domont, Gilberto B.; Santos, Keity S.; Zatz, Mayana; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); Instituto de Investigação em Imunologia-Instituto Nacional de Ciências e Tecnologia-iii-INCT; Necker Hospital for Sick Children; University of Paris; The Rockefeller University; Federal University of Rio de Janeiro ItemThe oldest unvaccinated Covid-19 survivors in South America(2022-12-01) de Castro, Mateus V.; Silva, Monize V. R.; Naslavsky, Michel S.; Scliar, Marilia O.; Nunes, Kelly; Passos-Bueno, Maria Rita; Castelli, Erick C. [UNESP]; Magawa, Jhosiene Y.; Adami, Flávia L.; Moretti, Ana I. S.; de Oliveira, Vivian L.; Boscardin, Silvia B.; Cunha-Neto, Edecio; Kalil, Jorge; Jouanguy, Emmanuelle; Bastard, Paul; Casanova, Jean-Laurent; Quiñones-Vega, Mauricio; Sosa-Acosta, Patricia; de Guedes, Jéssica S.; de Almeida, Natália P.; Nogueira, Fábio C. S.; Domont, Gilberto B.; Santos, Keity S.; Zatz, Mayana; Universidade de São Paulo (USP); Universidade Estadual Paulista (UNESP); Instituto de Investigação em Imunologia-Instituto Nacional de Ciências e Tecnologia-iii-INCT; Necker Hospital for Sick Children; University of Paris; The Rockefeller University; Federal University of Rio de JaneiroBackground: Although older adults are at a high risk of severe or critical Covid-19, there are many cases of unvaccinated centenarians who had a silent infection or recovered from mild or moderate Covid-19. We studied three Brazilian supercentenarians, older than 110 years, who survived Covid-19 in 2020 before being vaccinated. Results: Despite their advanced age, humoral immune response analysis showed that these individuals displayed robust levels of IgG and neutralizing antibodies (NAbs) against SARS-CoV-2. Enrichment of plasma proteins and metabolites related to innate immune response and host defense was also observed. None presented autoantibodies (auto-Abs) to type I interferon (IFN). Furthermore, these supercentenarians do not carry rare variants in genes underlying the known inborn errors of immunity, including particular inborn errors of type I IFN. Conclusion: These observations suggest that their Covid-19 resilience might be a combination of their genetic background and their innate and adaptive immunity.