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BACKGROUND The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) B.1.1.33-derived lineage named N.9 was described recently in Brazil and it’s considered a potential variant of interest (VOI) due to the presence of E484K substitution at the receptor-binding domain (RBD) of the Spike (S) protein. OBJECTIVE To describe the first detection of variant N.9 in Rio de Janeiro State. METHODS SARS-CoV-2 N.9 was confirmed by quantitative reverse transcription polymerase chain reaction (qRT-PCR), whole-genome sequencing and phylogenetic analysis. FINDINGS Here, we report two SARS-CoV-2 N.9 lineage strains in Rio de Janeiro. One of them had only the E484K substitution of the six N.9 lineage-defining mutations. Other three strains pre-defined as N.9 have the same genomic profile. These four strains are grouped within the B.1.1.33 lineage and basal to the N.9 lineage in our phylogenetic analysis, and we call them “N.9-like/B.1.1.33 + E484K”. MAIN CONCLUSIONS The phylogenetic analysis shows four independent introductions of N.9 in the state of Rio de Janeiro in October and December 2020, January and March 2021. SARS-CoV-2 N.9 dissemination in the Rio de Janeiro could have been limited by the emergence and dominance of other variants, mainly by the lineage P.2 VOI Zeta that emerged in the same period and co-circulated with N.9, as observed in the neighboring State of São Paulo.

O Hemocentro de Ribeirão Preto possui longa experiência na manufatura de células em Terapia Celular, através de uma infraestrutura com padrão de boas práticas que segue os critérios exigidos pela regulamentação brasileira. Recentemente, o Hemocentro expandiu seu foco para a produção de células T geneticamente modificadas (células CAR-T), através da validação do processo de manufatura de células CAR-T anti-CD19. Objetivo: Demonstrar a estruturação e validação de uma plataforma brasileira para a manufatura de células CAR-T anti-CD19 no cenário brasileiro. Material e métodos: Para a estruturação da plataforma, uma linha de cuidados foi estabelecida em 04 etapas de acordo com o processo de manufatura. A etapa um, consistiu na coleta de leucócitos totais em sangue periférico por procedimento de aférese seguida de isolamento das células mononucleares (CMN) por meio de Ficoll Hypaque e, posteriormente, congeladas para permanência em estoque até o momento do processamento. Na etapa dois, as CMN foram descongeladas para a depleção de monócitos após o qual, as células-T foram ativadas por meio de Dynabeads CD3/CD28. Na etapa três (24 horas após a etapa dois), as células-T foram transduzidas com vetor lentiviral e expandidas em meio AIMV (com 5% de soro humano AB e IL-2) por aproximadamente 8 a 11 dias. A última etapa, consistiu na execução dos seguintes testes de liberação e controle de qualidade: identidade/pureza, teste funcional/potência, análise microbiológica, análise do nível de endotoxinas bacterianas, teste para detecção de Mycoplasma, análise citogenética, viabilidade e contagem de beads residuais. Resultados: Até o momento, foram produzidos 11 lotes de células CAR-T que foram expandidas até atingir a dose mínima de 1,0 x 106 CAR-T/kg de peso do paciente e, após criopreservadas em bolsas. A mediana do perfil imunofenotípico dos 11 lotes de células CAR-T produzidos foi CD3 94,25%, CD4 38,1%, CD8 56,82%, CD16/56 2%, CD45/14 0,2%, CD19 0,2% e a % de CAR-CD19+ 35,8%. Em todos os lotes de células CAR-T produzidas houve lise das células alvos CD19+ evidenciado através de teste de potência. Os ensaios de liberação e do controle de qualidade da etapa 04 mostraram-se todos adequados para qualificar o produto final como apto para o uso clínico. Os lotes de células CAR-T apresentaram uma mediana de viabilidade de 91,81%. A contagem de beads residuais em todos os lotes foi inferior a 100 beads para cada 3x106 células. Discussão: Todos os lotes de células CAR-T CD19+ apresentaram-se com perfil imunofenotípico, atividade funcional, ensaios de controle de qualidade e com critérios de aceitação adequados dentro dos padrões GMP e da RDC ANVISA 508 de 2021. Dentre os 11 lotes produzidos, três foram utilizados em cenário clínico em tratamento compassivo de pacientes portadores de linfoma não-hodgkin de células B com níveis aceitáveis de toxicidade pós-infusão. Conclusão: A plataforma de manufatura de células CAR-T desenvolvida em Ribeirão Preto mostrou-se viável, segura e adequada para uso clínico em cenário brasileiro. O desenvolvimento de uma estrutura nacional tem o potencial de impactar o avanço científico da terapia celular avançada brasileira e, além disso, permitir maior acesso dos pacientes a esta terapia que é reconhecidamente de altíssimo custo.

The blood ammonia (NH3) level is one of the most important hepatic biomarkers for the diagnosis and monitoring of liver pathologies and infections. In this work, we developed an optimized optical biosensing method to extract and quantify the ammonia contained in complex-matrix samples emulating the blood serum. First, the approach was tested with solutions of phosphate-buffered saline (PBS) and ammonia chloride. Then, further trials were carried out with solutions of fetal bovine serum (FBS). The ammonia was extracted from the tested samples through a customized cell, and it was optically quantified by exploiting the indophenol reaction. The extraction cell included a cation-exchange membrane in Nafion, which was chemically pre-treated through cleaning procedures of sulfuric acid and hydrogen peroxide to keep a basic pH in the ammonia solution and to avoid contaminants in the membrane. From the NH3 solution, the indophenol reaction produced light-reactive indophenol dye molecules, which were used as colorimetric indicators. Through absorbance measurements of the indophenol dye solution at 670 nm wavelength, we were able to detect and quantify the ammonia level in the samples both with a spectrophotometer and a customized miniaturized read-out system, obtaining a detection limit of 0.029 µmol/mL.

In approximately 15% of patients with acute myeloid leukemia (AML), total and phosphorylated EGFR proteins have been reported to be increased compared to healthy CD34+ samples. However, it is unclear if this subset of patients would benefit from EGFR signaling pharmacological inhibition. Pre-clinical studies on AML cells provided evidence on the pro-differentiation benefits of EGFR inhibitors when combined with ATRA or ATO in vitro. Despite the success of ATRA and ATO in the treatment of patients with acute promyelocytic leukemia (APL), therapy-associated resistance is observed in 5-10% of the cases, pointing to a clear need for new therapeutic strategies for those patients. In this context, the functional role of EGFR tyrosine-kinase inhibitors has never been evaluated in APL. Here, we investigated the EGFR pathway in primary samples along with functional in vitro and in vivo studies using several APL models. We observed that total and phosphorylated EGFR (Tyr992) was expressed in 28% and 19% of blast cells from APL patients, respectively, but not in healthy CD34+ samples. Interestingly, the expression of the EGF was lower in APL plasma samples than in healthy controls. The EGFR ligand AREG was detected in 29% of APL patients at diagnosis, but not in control samples. In vitro, treatment with the EGFR inhibitor gefitinib (ZD1839) reduced cell proliferation and survival of NB4 (ATRA-sensitive) and NB4-R2 (ATRA-resistant) cells. Moreover, the combination of gefitinib with ATRA and ATO promoted myeloid cell differentiation in ATRA- and ATO-resistant APL cells. In vivo, the combination of gefitinib and ATRA prolonged survival compared to gefitinib- or vehicle-treated leukemic mice in a syngeneic transplantation model, while the gain in survival did not reach statistical difference compared to treatment with ATRA alone. Our results suggest that gefitinib is a potential adjuvant agent that can mitigate ATRA and ATO resistance in APL cells. Therefore, our data indicate that repurposing FDA-approved tyrosine-kinase inhibitors could provide new perspectives into combination therapy to overcome drug resistance in APL patients.

Four new Colombian species of the spider genus Medionops Sánchez-Ruiz & Brescovit, 2017 are described and illustrated: M. carolinae sp. nov. (male and female) from Boyacá department, M. cauca sp. nov. (male) from Cauca department, M. luiscarlosi sp. nov. (male and female) from Caldas department and M. santarosa sp. nov. (male) from Risaralda department. Additionally, the distribution of the type species of M. blades Sánchez-Ruiz & Brescovit, 2017, previously known only from Bogotá, Cundinamarca department, is extended to several localities in the Boyacá department. An updated identification key for all Medionops species is provided.

Abstract The aim of the study was to identify sources of resistance of Passiflora rootstocks to Fusarium sp. in two experimental areas with histories of disease foci in Mato Grosso state, Brazil. The experiment was carried out from June 2012 to December 2015 and was of randomized block design with eight treatments in which susceptible cultivar P. edulis was grafted onto hybrids CPAC MJ-H-87, CPAC MJ-H-76, CPAC MJ-H-86 and CPAC MJ-H-88, the commercial cultivar and native P. edulis, P. nitida and P. alata. Mortality rates of grafted plants were evaluated up to 640 days after transplantation. All Fusarium sp. isolates were identified as F. oxysporum f. sp. passiflorae. Plant mortality occurred earlier in the clay area than in the area with sandy clay loam. Grafted plants involving P. edulis as rootstocks were highly susceptible to Fusarium sp. with overall mortality rates above 56.25% considering both soil types, while plants with CPAC MJ-H-76 or native P. nitida rootstocks were more resistant with mortality rates below 12.5%. Grafted plants with CPAC MJ-H-86 as rootstock were highly resistant in clay soil but highly susceptible in soil with high proportion of sand. Only grafted plants involving CPAC MJ-H-76 as rootstock showed moderate resistance.

We propose a new point-prediction model, the DEep Learning WAVe Emulating model (DELWAVE), which successfully emulates the behaviour of a numerical surface ocean wave model (Simulating WAves Nearshore, SWAN) at a sparse set of locations, thus enabling numerically cheap large-ensemble prediction over synoptic to climate timescales. DELWAVE was trained on COSMO-CLM (Climate Limited-area Model) and SWAN input data during the period of 1971–1998, tested during 1998–2000, and cross-evaluated over the far-future climate time window of 2071–2100. It is constructed from a convolutional atmospheric encoder block, followed by a temporal collapse block and, finally, a regression block. DELWAVE reproduces SWAN model significant wave heights with a mean absolute error (MAE) of between 5 and 10 cm , mean wave directions with a MAE of 10–25°, and a mean wave period with a MAE of 0.2 s. DELWAVE is able to accurately emulate multi-modal mean wave direction distributions related to dominant wind regimes in the basin. We use wave power analysis from linearised wave theory to explain prediction errors in the long-period limit during southeasterly conditions. We present a storm analysis of DELWAVE, employing threshold-based metrics of precision and recall to show that DELWAVE reaches a very high score (both metrics over 95 %) of storm detection. SWAN and DELWAVE time series are compared to each other in the end-of-century scenario (2071–2100) and compared to the control conditions in the 1971–2000 period. Good agreement between DELWAVE and SWAN is found when considering climatological statistics, with a small (≤ 5 %), though systematic, underestimate of 99th-percentile values. Compared to control climatology over all wind directions, the mismatch between DELWAVE and SWAN is generally small compared to the difference between scenario and control conditions, suggesting that the noise introduced by surrogate modelling is substantially weaker than the climate change signal. [ABSTRACT FROM AUTHOR]

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ABSTRACT Advances in nanotechnology, combined with the use of natural products, represent a promissing research field. Brazil is a country of a rich biodiversity, especially in the Amazon forest. Fruits commonly used by local communities, such as bacaba (Oenocarpus bacaba), are potentially important for prospection of industrial applications of natural products. In nanotechnology, microemulsions stand out for providing a modified release to conveyed substances. This work aimed to develop microemulsionated formulations of bacaba oil, characterize them and evaluate their stability. We determined the HLB (hydrophile-lipophilic balance) of bacaba oil for formulation development. Six formulations were selected from pseudoternary diagrams, which indicated the proportions of surfactants, aqueous phase and bacaba oil. The viability of these formulations was evaluated through stability tests. We provided the rheological characterization of the formulations, evaluated their potential antioxidant activity through the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging method, and determined the hydrodynamic diameter of the droplets. The microemulsions were stable throughout the test period. Droplet diameter was below 200 nm, and the microemulsions were characterized as newtonian fluids, presenting an increase in antioxidant activity when compared with the diluted oil. Our results confirm the potential of bacaba oil in microemulsionated formulation as a suitable carrier for active compounds.

ABSTRACTThe yellow fever virus (YFV) caused a severe outbreak in Brazil in 2016–2018 that rapidly spread across the Atlantic Forest in its most populated region without viral circulation for almost 80 years. A comprehensive entomological survey combining analysis of distribution, abundance and YFV natural infection in mosquitoes captured before and during the outbreak was conducted in 44 municipalities of five Brazilian states. In total, 17,662 mosquitoes of 89 species were collected. Before evidence of virus circulation, mosquitoes were tested negative but traditional vectors were alarmingly detected in 82% of municipalities, revealing high receptivity to sylvatic transmission. During the outbreak, five species were found positive in 42% of municipalities. Haemagogus janthinomys and Hg. leucocelaenus are considered the primary vectors due to their large distribution combined with high abundance and natural infection rates, concurring together for the rapid spread and severity of this outbreak. Aedes taeniorhynchus was found infected for the first time, but like Sabethes chloropterus and Aedes scapularis, it appears to have a potential local or secondary role because of their low abundance, distribution and infection rates. There was no evidence of YFV transmission by Aedes albopictus and Aedes aegypti, although the former was the most widespread species across affected municipalities, presenting an important overlap between the niches of the sylvatic vectors and the anthropic ones. The definition of receptive areas, expansion of vaccination in the most affected age group and exposed populations and the adoption of universal vaccination to the entire Brazilian population need to be urgently implemented.

Yellow fever virus (YFV) caused an outbreak in the Brazilian Southeast from 2016 to 2019, of the most significant magnitude since the 1900s. An investigation of the circulating virus revealed that most of the genomes detected in this period carried nine unique amino acid polymorphisms, with eight located in the non-structural proteins NS3 and NS5, which are pivotal for viral replication. To elucidate the effect of these amino acid changes on viral infection, we constructed viruses carrying amino acid alterations in NS3 and NS5, performed infection in different cells, and assessed their neurovirulence in BALB/c mice and infected AG129 mice. We observed that the residues that compose the YFV 2016–2019 molecular signature in the NS5 protein might have been related to an attenuated phenotype, and that the alterations in the NS3 protein only slightly affected viral infection in AG129 mice, increasing to a low extent the mortality rate of these animals. These results contributed to unveiling the role of specific naturally occurring amino acid changes in the circulating strain of YFV in Brazil.

A 121-day feeding trial was undertaken to test the effects of two dietary lipid levels (16 and 21% L16, L21) in triplicated gilthead sea bream groups (initial weight: 67.5 g) reared at two different water temperatures (high, H 23°C and low, L 17°C) in the same recirculation system but exposed to a switch in temperature after 58 days. Fish kept at H were transferred to L (HL transition, autumn shift), and the fish kept at L were exposed to H (LH transition, summer shift), while continuing to receive the same diet to apparent satiation in each group. At the end of the trial, no significant diet effect on specific growth rate (SGR), feed intake (FI), and feed conversion rate (FCR) were detected in fish exposed to HL transition compared with those exposed to LH transition, while gross lipid efficiency (GLE) and lipid efficiency ratio (LER) were higher in L16. After temperature changes, L16 displayed higher SGR, FI, GLE, and LER, while mesenteric fat index was reduced. After temperature changes, the combined effects of low lipid diet and low temperature conditions resulted in higher pepsin activity, while trypsin, chymotrypsin, and lipase activities were generally higher at high lipid content. The combined effect of diet and temperature did not alter the metabolic plasma profile, except for the observed final higher aspartate aminotransferase (AST) and alkaline phosphatase (ALP) values when combining high dietary lipid (L21) and temperature changes. Different diets showed a significantly different gut microbiome layout, only at high temperature with L16 diet resulting in a higher load of Lactobacillus. On the contrary, no dietary impact on ecosystem diversity was observed, independently from the temperature. In addition, L16 diet in the HL transition favored an increase in Weissella and Bradyrhizobium genera in the gut microbiome, while in the final condition of LH transition, L21 diet favored a significant increase in Streptococcus and Bacillus. According to the results, the utilization of 16% dietary lipid levels in gilthead sea bream should be preferred during seasonal temperature changes in order to optimize feed utilization and gut health.