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Design, synthesis, and pharmacological evaluation of nitroimidazooxazine derivatives as inhibitors of cysteine protease B (CPB) from Leishmania spp.
No full textAs leishmanioses são uma doença parasitária causadas por mais de 20 espécies diferentes de protozoários do gênero Leishmania. Os maiores inconvenientes para o tratamento da doença é a limitada disponibilidade de fármacos, juntamente com os efeitos colaterais provocados, com isto, torna-se necessário a busca por novas agentes terapêuticos. Devido a problemática apontada, o grupo de pesquisa LAPDESF, em parceria com o grupo InovaLeish, vem trabalhando no planejamento e síntese de nova classe de compostos que atuem na inibição da enzima cisteíno protease B, sendo esta essencial para sobrevivência de espécies de Leishmania. Foram realizados ensaios nas formas amastigota de L. amazonensis (La) e L. infantum (Li), agentes responsáveis pela leishmaniose cutânea e visceral, respectivamente. A síntese de nove derivados peptídeomiméticos (PEP01-PEP10) demonstraram potente atividade contra CPB com destaque para os derivados PEP01 e PEP02, com valores de inibição de 4,3 e 0,8 μM. Além disso, o composto PEP01 apresentou atividade anti-leishmanial para L. amazonensis (4,6 μM) e L. infantum (3,9 μM). Com base nos resultados apresentados, os grupos LAPDESF e Inovaleish, vêm identificando novas alternativas para o tratamento da leishmaniose.Financiadora de Estudos e Projetos (FINEP)FINESP: 3348/2022 - CC
Genetic diversity in wild and breeding populations and clones of <i>Eucalyptus urophylla</i> and <i>Eucalyptus grandis</i>
No full textBrazil has a long history of intensive silviculture with Eucalyptus species, mainly Eucalyptus urophylla and E. grandis. However, breeding advances may reduce genetic diversity in bred populations. Nine microsatellite markers assessed genetic diversity in wild and improved populations of E. urophylla and E. grandis, and genetic similarity in nine widely planted clones. Four wild populations of E. urophylla were evaluated: Flores, Timor-Leste, Timor and other Islands, along with three improved populations. For E. grandis, one wild and one improved population were analyzed. Results showed higher genetic diversity in improved populations, possibly due to admixed composition from different provenances. Wild populations of E. urophylla formed two distinct groups. All clones were genetically similar to improved E. urophylla populations. Some clones are inter-specific hybrids, contradicting their reported pedigree, but predominantly E. urophylla.Inst Pesquisa & Estudos Florestais IPEF, Via Comendador Pedro Morganti 3500, BR-13415000 Piracicaba, SP, BrazilUniv Sao Paulo, Escola Super Agr Luiz Queiroz, Ave Padua Dias 11,CP 9, BR-13418900 Piracicaba, SP, BrazilInst Florestal Sao Paulo, CP 1322, BR-01059970 Sao Paulo, SP, BrazilUNESP, Fac Engn Ilha Solteira, Ilha Solteira Campus, Ilha Solteira, SP, BrazilAbacus Bio, Ground Floor,Publ Trust Bldg,442 Moray Pl, Dunedin 9010, New ZealandUniv Fed Lavras, Ave Cent S-N Campus Univ, BR-37200900 Lavras, MG, BrazilSuzano Papel & Celulose, Unidade Mucuri, BR 101,Km 945,5,Caixa Postal 63, BR-45930000 Mucuri, BA, BrazilUniv Sunshine Coast, Locked Bag 4,Maroochydore DC, Sippy Downs, Qld 4558, AustraliaEMBRAPA Genet Resources & Biotechnol, Plant Genet Lab, BR-70770970 Brasilia, DF, BrazilGrad Program Genom Sci Biotechnol, SGAN Qd 916, BR-70790160 Brasilia, DF, BrazilUniv Catol Brasilia, SGAN Qd 916, BR-70790160 Brasilia, DF, BrazilUNESP, Fac Engn Ilha Solteira, Ilha Solteira Campus, Ilha Solteira, SP, Brazi
Greener Epoxidation Reaction of Macaw Palm Oil Using Metal Oxides and Niobium Phosphate as Catalysts
No full textThe present work focused on the screening process of different inorganic oxides and different temperatures for the epoxidation reaction of macaw palm vegetable oil without using solvent, a promising Brazilian biomass that can substitute other vegetable oils (e.g., soybean oil) in technological applications. This oil has an iodine value of 108.48 g of I2 per 100 g being categorized as a semi-dry oil. The epoxidized vegetable oil can be applied in manifold areas in polymer science. The epoxidation reaction here presented avoided organic solvents and organic acids and aimed to produce a greener route using metal oxides or niobium phosphate as a peroxide-activating catalyst. The experimental parameters were maintained as 10 mol% of catalyst loading and a time of 24 h; however, the temperature for some catalysts was changed to improve the epoxide conversion. Experiments were measured by proton nuclear magnetic resonance (1H NMR). Experiments using titanium oxide (TiO2) as a catalyst reached a conversion greater than 84.7%, while experiments using aluminium oxide (Al2O3) or lanthanum oxide (La2O3) achieved conversions of 22.6 and 30.1%, respectively. Furthermore, it was observed that both niobium compounds instantly react with hydrogen peroxide to form peroxy derivatives.Laboratório de Análise Térmica Eletroanalítica e Química de Soluções (LATEQS) Instituto de Química de São Carlos Universidade de São Paulo (USP), SPLaboratório de Análise Térmica e Polímeros (LATP) Departamento de Química Faculdade de Ciências Universidade Estadual Paulista (Unesp), SPLaboratório de Análise Térmica e Polímeros (LATP) Departamento de Química Faculdade de Ciências Universidade Estadual Paulista (Unesp), S
Tecnologias para obter sementes de amendoim com qualidade superior: do campo ao gene
No full textThe quality of peanut seeds is essential for optimized germination, storage and the rapid, uniform establishment of seedling in the field (vigor), making it crucial for agriculture. This study explores innovative methods associated with the physiological quality of peanut seeds at different development stages and in the post-harvest phase, with approaches focusing on maturation, chemical composition, spectroscopy techniques, and gene expression. In Chapter 1, the effectiveness of combining multispectral imaging with artificial intelligence was investigates to autonomously and non-destructively identify seedlots with superior vigor. New markers of seed and seedling performance, such as anthocyanin, were identified, opening new perspectives for seed vigor analysis. In Chapter 2, the challenge of determining the ideal harvest time for peanut seeds in the field was addressed. Focusing on the maturation and late maturation phases, this research proposes a novel maturation scale for peanut in tropical agriculture, providing practical guidelines for harvesting peanut seeds with superior germination, desiccation tolerance, vigor, longevity, and health (seed quality attributes). In Chapter 3, the chemical composition of seeds at different development stages was explored, examining whether non-destructive techniques commonly used in other sectors (e.g., LIBS in the mining industry) can identify chemical patterns associated with mature seeds. Techniques such as Nuclear Magnetic Resonance, Laser-Induced Breakdown Spectroscopy (LIBS), and X-ray Fluorescence were effective in identifying seeds with superior vigor through the digital dynamics of oil and minerals. Finally, in Chapter 4, the expression of genes associated with flavonoid biosynthesis, specifically the anthocyanin pathway, was investigated, along with their possible involvement in the acquisition of the seed quality attributes. The secondary metabolism of peanuts was explored during seed maturation phases, revealing contributions to the species' biology and biotechnological applications in Brazilian agriculture. Together, these four studies provide a comprehensive understanding of the physiological, chemical, spectral and molecular aspects of peanut seeds, as well as valuable insights for the technological advancement of this crop production system in Brazil and worldwide.A qualidade das sementes de amendoim é essencial para uma germinação otimizada, armazenamento e estabelecimento rápido e uniforme das plântulas no campo (vigor), sendo crucial para a agricultura. Este estudo explora métodos inovadores associados à qualidade fisiológica das sementes de amendoim em diferentes estádios de desenvolvimento e na pós-colheita, com abordagens sobre a maturação, composição química, técnicas de espectroscopia e expressão gênica. No Capítulo 1, a eficácia da combinação de imagens multiespectrais com inteligência artificial foi investigada para identificar de forma autônoma e não destrutiva lotes de sementes com qualidade superior. Marcadores inéditos do desempenho de sementes e plântulas, como a antocianina, foram encontrados abrindo novas perspectivas para a análise do vigor de sementes. No Capítulo 2, foi abordado o desafio de determinar o momento ideal para a colheita de sementes de amendoim no campo. Focando nas fases de maturação e maturação tardia, esta pesquisa propõe uma escala de maturação inédita para o
amendoim na agricultura tropical, fornecendo diretrizes práticas para a colheita de sementes com germinação, tolerância à dessecação, vigor, longevidade e sanidade superiores (atributos da qualidade). No Capítulo 3, explorou-se a composição química das sementes nos diferentes estádios de desenvolvimento, examinando se técnicas não destrutivas comuns em outros setores (ex: LIBS na indústria de mineração) podem identificar padrões químicos associados às sementes de amendoim maduras. Técnicas como Ressonância Magnética Nuclear, Espectroscopia de Ruptura Induzida por Laser (LIBS) e Fluorescência de Raios X foram eficazes para identificar sementes com vigor superior por meio da dinâmica digital de óleo e minerais. Por fim, no Capítulo 4, foi investigado a expressão dos genes associados a biossíntese de flavonoides, rota das antocianinas, e o possível envolvimento deles com a aquisição dos componentes de qualidade das sementes. O metabolismo secundário do amendoim foi explorado durante as fases de maturação, revelando contribuições à biologia de sementes da espécie e aplicações biotecnológicas na agricultura brasileira. Juntos, esses quatro estudos fornecem uma compreensão abrangente sobre os aspectos fisiológico, químico, espectral e molecular das sementes de amendoim além de valiosos insights para o progresso tecnológico do sistema produtivo da cultura no Brasil e no mundo.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPESP: 2020/14050-3FAPESP: 2023/00435-9CNPq; 142236/2020-
Primary cutaneous cryptococcosis – History, concepts, clinical and therapeutic update
No full textCryptococcosis is a disease caused by fungi of the genus Cryptococcus, with the species Cryptococcus neoformans and Cryptococcus gattii being recognized as pathogenic. Cutaneous cryptococcosis can be classified as “secondary”, developing from a previous systemic disease, or, on the contrary, “primary”, resulting from transcutaneous inoculation of the agent. It can also be classified as “disseminated cutaneous cryptococcosis”, when there is an associated systemic disease, or “localized”, when it is restricted to the skin. This article uses the term “primary cutaneous cryptococcosis” because it is the most widely used and already established in the literature. Historically, the first report of a possible case of primary cutaneous cryptococcosis (PCC) occurred in 1950 by Gancy WM and was published in the Archives of Dermatology and Syphilology. Subsequently, the rare and sporadic reports in the following decades were reviewed and reported in the 1985 publication by Baes & van Cutsen. However, the unequivocal acceptance of the existence of PCC as a distinct disease only occurred in 2003 with the publication by Neville S et al. of the French Cryptococcosis Study Group. The fundamental criterion established to consider it as PCC was the proven absence of systemic disease, whether pulmonary, in the CNS or other location at the time of diagnosis of the cutaneous condition, characterized by a single lesion and, mostly, in an exposed area. These and other clinical criteria, diagnostic confirmation, and therapeutic choice are discussed in detail in the full text.Department of Infectology Dermatology Diagnostic Imaging and Radiotherapy Faculty of Medicine Universidade Estadual Paulista, SPDepartment of Infectology Dermatology Diagnostic Imaging and Radiotherapy Faculty of Medicine Universidade Estadual Paulista, S
Macrophyte species uniqueness is driven by habitat integrity, sediment structure, and spatial components in streams around the Amazon National Park
No full textNatural landscapes have been altered due to the increasing demand for natural resources, leading to changes in physicochemical characteristics and species composition of several systems, including the aquatic. This study identified the streams and species that contributed the most to macrophyte beta diversity within and surrounding the Amazon National Park. We also investigated the factors (local, spatial, and land-use) influencing the community structure. Biological and environmental data were obtained by field sampling in 29 streams (17 within the park and 12 outside). Land-use data were obtained by remote sensing (satellite imagery). The highest species contribution to beta diversity was observed in species with a high frequency of occurrence. The greater local contribution to beta diversity (LCBD) and the highest species richness were observed outside the park. Habitat integrity, fine substrate, and spatial components (i.e., geographic distance) negatively influenced LCBD, highlighting the effect of environmental filtering and dispersion processes on stream macrophytes. Land use variables did not significantly affect macrophytes LCBD. Our findings provide an important snapshot of how different factors (local, spatial, and land use changes) interact in determining macrophyte beta diversity in streams within and around the Amazon National Park. The study also underscores the importance of conservation units to preserve stream ecosystems' natural characteristics and habitat integrity.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Foundation Fighting BlindnessConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Programa de Pós-Graduação em Ecologia Instituto de Ciências Biológicas Universidade Federal do Pará- UFPA, Av. Perimetral, 1 - Guamá, ParáPrograma de Pós-Graduação em Zoologia da Universidade Federal do Pará- UFPA, Av. Perimetral, 1 - Guamá, ParáInstituto de Biociências Letras e Ciências Exatas Universidade Estadual Paulista ‘Júlio de Mesquita Filho’ UNESP, R. Cristóvão Colombo, 2265, São PauloLaboratório de Estudos de Impacto Ambiental (LEIA) Instituto de Ciências e Tecnologia das Águas (ICTA) Universidade Federal do Oeste do Pará (UFOPA), ParáInstituto de Biociências Letras e Ciências Exatas Universidade Estadual Paulista ‘Júlio de Mesquita Filho’ UNESP, R. Cristóvão Colombo, 2265, São PauloFAPESP: 2019/25445-1Foundation Fighting Blindness: 2022/1437669CNPq: 302881/2022–0CNPq: 304403/2021–0CNPq: 304710/2019–9CNPq: 307808/2022–0CNPq: 311550/2023–1CNPq: 311835/2023–6CNPq: 428961/2018-
Action of the aqueous extract of cataia (Pimenta Pseudocaryophyllus) on the development of Spodoptera Eridania (Lepidoptera: noctuidae)
No full textA Spodoptera eridania, conhecida como lagarta-das-folhas, é um inseto polífago que se alimenta principalmente das culturas da soja, arroz, cana de açúcar, algodão e milho. O uso de extratos vegetais pode ser uma boa ferramenta no manejo de pragas agrícolas visando uma agricultura mais sustentável. O presente trabalho teve como objetivo avaliar a influência do extrato aquoso de cataia (Pimenta pseudocaryophyllus), incorporado à dieta artificial no desenvolvimento deste inseto. O delineamento experimental utilizado foi o inteiramente casualizado (DIC) com 5 tratamentos e 5 repetições, com cada repetição composta por 5 lagartas. Para a obtenção dos tratamentos de extrato aquoso foram utilizados 5, 10, 15 e 20g de pó vegetal, completando o volume do recipiente para os 100ml com água destilada. As lagartas recém eclodidas utilizadas foram oriundas de criação estoque mantida no laboratório de Fitossanidade da FCAVR, em câmara climatizada (BOD) com 14 h de fotofase, 25oC e UR de 70%, mesmas condições em que foi mantido o experimento. Os tratamentos foram compostos por diferentes concentrações de extrato aquoso da cataia, incorporados a dieta artificial que serviu de alimento para as lagartas, incorporando-se água destilada no tratamento controle. Foram aferidos os pesos larvais em 7, 9 e 11 dias após a exposição (DAE), sobrevivência e duração da fase larval, o consumo alimentar e o peso de fezes produzido pelas lagartas ao final do ciclo larval e peso, duração e sobrevivência da fase pupal. Na fase adulta avaliou-se a razão sexual e longevidade de machos e fêmeas avaliando-se ao final a duração do ciclo ovo-adulto. Os resultados indicaram que as maiores concentrações do extrato aquoso de P. pseudocaryophyllus afetaram negativamente o desenvolvimento de S. eridania, ocasionando redução no peso larval. Os tratamentos causaram prolongamento do período larval e consequentemente do período ovo-adulto. Por causar efeitos adversos na biologia de S. eridania, é importante a realização de novos estudos visando elucidar os compostos ativos de P. pseudocaryophyllus, bem como seu modo de ação, para possível uso em programas de manejo de pragas.Spodoptera eridania, known as the leafworm, is a polyphagous insect that feeds primarily on soybean, rice, sugarcane, cotton, and corn crops. The use of plant extracts can be a valuable tool in agricultural pest management, aiming for more sustainable agriculture. This study aimed to evaluate the influence of aqueous extract of cataia (Pimenta pseudocaryophyllus), incorporated into an artificial diet, on the development of this insect. A completely randomized experimental design (CRD) was used, with five treatments and five replicates, each replicate consisting of five caterpillars. Aqueous extract treatments were prepared using 5, 10, 15, and 20g of plant powder, and the container volume was replenished to 100ml with distilled water. The newly hatched caterpillars used were from stock reared in the FCAVR Plant Health Laboratory, in a climate-controlled chamber (BOD) with a 14-h photophase, 25°C, and 70% RH, the same conditions under which the experiment was conducted. The treatments consisted of different concentrations of aqueous extract of cataia, incorporated into an artificial diet that served as food for the caterpillars, with distilled water being incorporated in the control treatment. Larval weights were measured at 7, 9, and 11 days after exposure (DAE), survival and duration of the larval stage, food consumption and fecal weight produced by the caterpillars at the end of the larval cycle, and the weight, duration, and survival of the pupal stage. In the adult stage, the sex ratio and longevity of males and females were evaluated, evaluating the duration of the egg-to-adult cycle at the end. The results indicated that higher concentrations of the aqueous extract of P. pseudocaryophyllus negatively affected the development of S. eridania, resulting in reduced larval weight. The treatments prolonged the larval period and, consequently, the egg-to-adult period. Because these treatments adversely affect the biology of S. eridania, further studies are needed to elucidate the active compounds of P. pseudocaryophyllus and their mode of action, for potential use in pest management programs
Seasonal dynamics of gastrointestinal helminths in naturally infected horses in Brazil
No full textHelminthiasis in horse breeding stands out due to the damage caused by this infection, which is mainly controlled by anthelmintic drugs. Knowing the population dynamics of helminths in a given region is therefore essential, as environmental conditions have a major impact on the development of these parasites in pastures. The aim of this study was to assess the influence of seasonality on helminths. It was carried out between October and September of the years 2015/2016, 2018/2019 and 2021/2022. A total of 147 animals were used, 64 females and 83 males, all from the same herd. Fecal samples were collected directly from the rectal ampulla of each animal every 28 days for 12 months of each cycle for coproparasitological analysis. Average air temperature, average relative humidity and rainfall were measured daily. The weather records were collected by the Agroclimatological Station in Castilho, SP (21o 05' 12.4 S; 51o 35' 17.2 W), located six kilometers from the experiment site. In the three study cycles, the EPG and rainfall averages showed significant differences between the times of year. In general, the month with the highest EPG in the different cycles was February, with 1626 in 2015/2016, 926.7 in 2018/2019 and 1885 in 2021/2022, and the month with the lowest average EPG was July, with 642 in 2015/2016, 610.5 in 2018/2019 and 804 in 2021/2022. Regarding climatic conditions, rainfall was more concentrated in two seasons, spring and summer, meaning that summer was the season with the highest rainfall in the three cycles evaluated. Autumn, followed by winter were the seasons with the lowest rainfall during the cycles studied. It can be concluded that climatic conditions have a direct influence on the population dynamics of helminths, and that during the drier and colder seasons, horses were less affected, so the suggestion for a possible strategic program would be to concentrate the administration of anthelmintics in spring and summer.Universidade Estadual Paulista (UNESP) School of Agrarian and Technological Sciences (FCAT), SPUniversidade Estadual Paulista (UNESP) School of Agrarian and Technological Sciences (FCAT), S
Are mixed models more effective than linear models in predicting energy utilization for laying hens?
No full textThis study was conducted to develop mathematical models that can overcome the limitations of linear energy partitioning models in laying hens comparing the metabolizable and net energy bases. Three linear models and 2 nonlinear mixed-effect models were fitted to predict the energy utilization efficiency for body tissue deposition and egg production, as well as the maintenance requirements for metabolizable energy (MEm) and net energy (NEm). A total of 30 diets were individually formulated to achieve values below and above the nutritional recommendation of the genetic guidelines. Heat production and energy metabolism were measured in laying hens during the production phase using 6 open-circuit respiration chambers. Diets were randomly assigned into 5 groups, and each diet was replicated 4 times with 6 hens per replication. The experimental protocol included a 5-d adaptation to the experimental diets, 2-d chamber adaptation, 4-d measurements of gas exchange, total excreta collection and growth performance recording under feeding conditions, and 1-d gas exchange measurement under fasting conditions. Feed intake, body weight, egg production, egg mass, and excreta production were measured daily. The variables for total heat production (THP) and fasting heat production (FHP) were obtained through the Brouwer equation using VO2 and VCO2 measurements. Apparent metabolizable energy intake (AMEi) was determined by the difference between the gross energy intake and excreted. Retained energy (RE) was determined as the difference between AMEi and THP. Subsequently, the RE was partitioned between RE in body and egg. The RE in egg and body were partitioned into RE as protein and fat. Statistical analyses involved linear regressions and nonlinear mixed-effect regressions of the main variables described. The values obtained for MEm, NEm, and the efficiency of energy retention in body (kbody), and egg (kegg) in the linear models were 106 kcal/kg0.75.d, 90 kcal/kg0.75.d, 0.843, and 0,779, respectively. For nonlinear models the values were 94.15 kcal/kg0.75.d, 81.72 kcal/kg0.75.d, 0.463, and 0.638 for MEm, NEm, kbody and kegg, respectively. Nonlinear models were considered the ideal choice to determine MEm and NEm requirements due to the lowest error. The non-linear mixed model provides a more accurate representation of energy utilization in laying hens compared to linear models, offering a better characterization of energy partitioning within the net energy bases.Department of Animal Science São Paulo State University School of Agricultural and Veterinary Sciences JaboticabalSchool of Agriculture and Environment Massey UniversityDepartment of Animal Science São Paulo State University School of Agricultural and Veterinary Sciences Jaboticaba
Evaluation of Grape Pomace Supplementation in Lamb Diets to Mitigate Haemonchus contortus Infection
No full textThis study aimed to evaluate the potential benefits and feasibility of adding dried and ground grape pomace to the concentrate fed to lambs experimentally infected with Haemonchus contortus. Eighteen male Santa Inês lambs, recently weaned, were housed in individual pens and were allocated into two groups based on their body weight. The lambs in the supplemented group (n = 9) initially received a diet composed of 50% ground hay and 50% concentrate. Subsequently, these animals were gradually adapted to grape pomace until its final inclusion in the concentrate reached 20%. The lambs in the control group received a concentrate without grape pomace. Both groups of lambs were artificially infected with 4000 infective larvae of H. contortus, and 28 days later, the lambs were euthanized for quantification of the nematodes present in the abomasum. The following variables showed no statistical differences (p > 0.05) between the groups: worm burden, packed cell volume, total plasma protein, blood eosinophil count, and daily weight gain. Regarding anti-Haemonchus IgG plasma levels, there was a significant time * treatment interaction (p = 0.0099) with higher values in the supplemented group. At the two final samplings, the supplemented group showed significantly lower values of eggs per gram of feces than the control group (p < 0.05). The supplemented group showed female worms shorter and with less eggs in utero than those of the control group, with significant difference for these variables (p < 0.05). In conclusion, grape pomace can be included in the diet, as it promotes more sustainable animal production, and, additionally, it can cause a reduction in H. contortus fecundity, benefiting haemonchosis prophylaxis.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Faculty of Veterinary Medicine and Animal Science São Paulo State University (UNESP), SPDepartment of Biodiversity and Biostatistics Institute of Biosciences UNESP, SPLuiz de Queiroz College of Agriculture (ESALQ) University of São Paulo (USP), SPAnimal Nutrition Laboratory Center for Nuclear Energy in Agriculture USP, SPFaculty of Agricultural and Technological Sciences UNESP, SPDepartment of Pathology Reproduction and One Health UNESP, SPFaculty of Veterinary Medicine and Animal Science São Paulo State University (UNESP), SPDepartment of Biodiversity and Biostatistics Institute of Biosciences UNESP, SPFaculty of Agricultural and Technological Sciences UNESP, SPDepartment of Pathology Reproduction and One Health UNESP, SPFAPESP: 2021/03479-