Purification of antileukemic drugs through silica-based supported ionic liquids

L-asparaginase (LA) is an enzyme used as a biopharmaceutical for the treatment of acute lymphoblastic leukemia. LA can be produced via fermentation and its purification usually comprises several steps including precipitation, liquid-liquid extraction and chromatography techniques. Among these, ion exchange chromatography, which is often preceded by precipitation with salts as a first pre-chromatographic step, is the most used. However, theses common strategies for protein purification result in low yields and purity, requiring long processing times, while leading to a consequent increase of the process costs. Therefore, the demand for new cost-effective production/purification processes play now a priority role. This work aims the development of cost-effective technologies to purify LA from the complex fermentation medium from Bacillus Subtillis. Silica-based supported ionic liquids (SILs) are investigated as cost-effective purification materials for the target enzyme. The concentration of the extract from the fermentation, material/ extract from fermentation ratio and contact time effects in the purity and yield of LA were optimized. With this strategy, process costs, energy consumed, and waste generated, may be significantly decreased, which may lead to this biopharmaceutical price decrease and wider application.publishe

Purification of antileukemic drugs through silica-based supported ionic liquids

L-asparaginase (LA) is an enzyme used as a biopharmaceutical for the treatment of acute lymphoblastic leukemia. LA can be produced via fermentation and its purification usually comprises ion exchange chromatography, which is often preceded by precipitation with salts as a first pre-chromatographic step. However, this purification strategy result in low yields and purity, requires long processing times, while leading to a consequent increase of the process costs. Therefore, the demand for new cost-effective purification processes play now a priority role. In this work silica-based supported ionic liquids (SILs) are investigated as an alternative technology to purify LA from the complex fermentation medium from Bacillus subtillis. The concentration of the extract from the fermentation, material/ extract from fermentation ratio and contact time effects in the purity and yield of LA were optimized. With this strategy, process costs, energy consumed, and waste generated, may be significantly decreased, which may lead to this biopharmaceutical price decrease and wider application.publishe

ESTRUTURA POPULACIONAL E CONDIÇÃO FISIOLÓGICA DE Hoplias malabaricus (BLOCH, 1794) NO BAIXO RIO CAPIBARIBE

This research aimed to determine population structure and physiological condition of Hoplias malabaricus in the Capibaribe river (PE), northeastern Brazil. Monthly sampling were carried out between September/2013 and August/2014, using gill nets. The sex was identified by macroscopic and microscopic viewing of gonads, following the classification proposed by Vazzoler (1996). The specimens were distributed in length classes, being applied Kolmogorov-Smirnov test to evaluate the presence of sexual dimorphism. The length-weight relationship was calculated assessing differences through Kruskal Wallis test and physiological condition was calculated through allometric condition factor (K) between sexes, being applied the Spearman correlation test. Was collected 30 males and 42 females. No sexual dimorphism was observed relative to the size. Was observed positive allometric growth for females and negative for males. Was evidenced physiological condition variations along the year regarding seasonality. Population structure and physiological condition of the Hoplias malabaricus differed from that found in other environments, being this research, an important tool support the development of future management plans and conservation of Hoplias malabaricus, especially in this region.Keywords: population dynamics; fishing; reproduction.Esta pesquisa objetivou determinar a estrutura populacional e condição fisiológica de Hoplias malabaricus no rio Capibaribe (PE), nordeste brasileiro. Foram realizadas coletas mensais entre setembro/2013 e agosto/2014, utilizando redes de espera. O sexo foi identificado pela visualização macroscópica e microscópica das gônadas, seguindo a classificação proposta por Vazzoler (1996). Os indivíduos foram distribuídos entre classes de comprimento, sendo aplicado teste de Kolmogorov-Smirnov para verificar a presença de dimorfismo sexual. A relação peso-comprimento entre sexos foi calculada, avaliando diferenças pelo teste Kruskal-Wallis e a condição fisiológica foi avaliada pelo fator de condição alométrico (K) entre sexos, sendo aplicado teste de correlação de Spearman. Foram coletados 30 machos e 42 fêmeas, não sendo observado dimorfismo sexual relativo ao tamanho. Além disto, verificou-se um crescimento alométrico positivo para as fêmeas e negativo para os machos. Também foram evidenciadas variações na condição fisiológica ao longo do ano em relação à sazonalidade. A estrutura populacional e condição fisiológica de Hoplias malabaricus diferiram do encontrado em outros ambientes, sendo este estudo uma importante ferramenta para embasar o desenvolvimento de futuros planos de manejo e conservação de Hoplias malabaricus, especialmente nesta região.Palavras-chave: dinâmica populacional, pesca, manejo de recursos pesqueiros

Novidade no mercado, novidade no ambiente: invasão do peixe não nativo acará onça no nordeste do Brasil

In Brazil, more than 150 non-native fish species have been introduced either intentionally or unintentionally, mainly due to aquaculture, fishery improvement and ornamental trade. The non-native jaguar guapote Parachromis managuensis was recorded in two artificial ponds, four rivers, one stream and two reservoirs, ecosystems belonging to the Una, Ipojuca, Capibaribe and Pajeú River basins, Pernambuco, northeastern Brazil. The sampling period was between October 2006 and July 2015. A total of 376 individuals of P. managuensis was captured by seines, casting nets and gillnets. This species may have been introduced in the study sites by accidental release during pond drainage in fish farms, stocking for the purpose of animal protein production or by aquarium dumping. The presence of various ontogenetic life stages in all studied environments is a strong evidence that the species is established. Seven management actions are proposed to prevent further introductions of P. managuensis in other environments of the region.Keywords: exotic, bioinvasion, introduction, aquarism, freshwater fish.No Brasil, mais de 150 espécies de peixes não nativos foram introduzidas intencionalmente ou não, especialmente devido à aquicultura, pesca e aquarismo. A presença do peixe não nativo Parachromis managuensis, o acará onça, foi registrada em dois açudes, quatro rios, um córrego e duas barragens distribuídos nas bacias hidrográficas dos rios Una, Ipojuca, Capibaribe e Pajeú, no estado de Pernambuco, nordeste do Brasil. O período de amostragem ocorreu entre outubro/2006 e julho/2015. Um total de 376 indivíduos de P. managuensis foi capturado utilizando tarrafas, redes de arrasto e redes de espera. Esse peixe pode ter sido introduzido nos locais estudados por escape acidental durante a drenagem de viveiros de piscicultura, estocagem com a finalidade de produção de proteína animal e através do descarte de aquários. A presença de vários estágios ontogênicos em todos os ambientes estudados representa uma forte evidência de que a espécie está estabelecida. Ações de manejo são propostas para evitar novas introduções de P. managuensis em outros ambientes da região.Palavras-chave: exótica, bioinvasão, introdução, aquarismo, peixes de água doce

Risk factors for infection, predictors of severe disease, and antibody response to COVID-19 in patients with inflammatory rheumatic diseases in Portugal: a multicenter, nationwide study

Copyright © 2022 Cruz-Machado, Barreira, Bandeira, Veldhoen, Gomes, Serrano, Duarte, Rato, Miguel Fernandes, Garcia, Pinheiro, Bernardes, Madeira, Miguel, Torres, Bento Silva, Pestana, Almeida, Mazeda, Cunha Santos, Pinto, Sousa, Parente, Sequeira, Santos, Fonseca and Romão. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.Objective: To identify risk factors for SARS-CoV-2 infection and for severe/critical COVID-19, and to assess the humoral response after COVID-19 in these patients. Methods: Nationwide study of adult patients with inflammatory RMDs prospectively followed in the Rheumatic Diseases Portuguese Register-Reuma.pt-during the first 6 months of the pandemic. We compared patients with COVID-19 with those who did not develop the disease and patients with mild/moderate disease with those exhibiting severe/critical COVID-19. IgG antibodies against SARS-CoV-2 were measured ≥3 months after infection and results were compared with matched controls. Results: 162 cases of COVID-19 were registered in a total of 6,363 appointments. Patients treated with TNF inhibitors (TNFi; OR = 0.160, 95% CI 0.099-0.260, P < 0.001) and tocilizumab (OR 0.147, 95% CI 0.053-0.408, P < 0.001) had reduced odds of infection. Further, TNFi tended to be protective of severe and critical disease. Older age, major comorbidities, and rituximab were associated with an increased risk of infection and worse prognosis. Most patients with inflammatory RMDs (86.2%) developed a robust antibody response. Seroconversion was associated with symptomatic disease (OR 13.46, 95% CI 2.21-81.85, P = 0.005) and tended to be blunted by TNFi (OR 0.17, 95% CI 0.03-1.05; P = 0.057). Conclusions: TNFi and tocilizumab reduced the risk of infection by SARS-CoV-2. Treatment with TNFi also tended to reduce rates of severe disease and seroconversion. Older age, general comorbidities and rituximab were associated with increased risk for infection and worse prognosis, in line with previous reports. Most patients with RMDs developed a proper antibody response after COVID-19, particularly if they had symptomatic disease.We acknowledge the generous sharing of the expression constructs by Dr. Florian Krammer, Icahn School of Medicine at Mount Sinai, New York, USA [Development of SARS-CoV-2 reagents was partially supported by the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract HHSN272201400008C] and the protein production by Drs. Paula Alves and Rute Castro at Instituto de Biologia Experimental e Tecnológica (iBET) Oeiras, Portugal as part of the Serology COVID consortium.info:eu-repo/semantics/publishedVersio

Anti-trbc1 antibody-based flow cytometric detection of t-cell clonality: Standardization of sample preparation and diagnostic implementation

© 2021 by the authors.A single antibody (anti-TRBC1; JOVI-1 antibody clone) against one of the two mutually exclusive T-cell receptor β-chain constant domains was identified as a potentially useful flow-cytometry (FCM) marker to assess Tαβ-cell clonality. We optimized the TRBC1-FCM approach for detecting clonal Tαβ-cells and validated the method in 211 normal, reactive and pathological samples. TRBC1 labeling significantly improved in the presence of CD3. Purified TRBC1+ and TRBC1− monoclonal and polyclonal Tαβ-cells rearranged TRBJ1 in 44/47 (94%) and TRBJ1+TRBJ2 in 48 of 48 (100%) populations, respectively, which confirmed the high specificity of this assay. Additionally, TRBC1+/TRBC1− ratios within different Tαβ-cell subsets are provided as reference for polyclonal cells, among which a bimodal pattern of TRBC1-expression profile was found for all TCRVβ families, whereas highly-variable TRBC1+/TRBC1− ratios were observed in more mature vs. naïve Tαβ-cell subsets (vs. total T-cells). In 112/117 (96%) samples containing clonal Tαβ-cells in which the approach was validated, monotypic expression of TRBC1 was confirmed. Dilutional experiments showed a level of detection for detecting clonal Tαβ-cells of ≤10−4 in seven out of eight pathological samples. These results support implementation of the optimized TRBC1-FCM approach as a fast, specific and accurate method for assessing T-cell clonality in diagnostic-FCM panels, and for minimal (residual) disease detection in mature Tαβ+ leukemia/lymphoma patients.This work was supported by the CB16/12/00400 (CIBERONC) and PI20-01346 grants, from the Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación (Madrid, Spain) and FONDOS FEDER, and by the EuroFlow Foundation (Leiden, The Netherlands). N. Muñoz-García is supported by a pre-doctoral grant (Ref. IBPredoc17/00012) from IBSAL (Salamanca, Spain). M. Lima, N. Villamor, A.W. Langerak, J.J.M. van Dongen, A. Orfao, and J. Almeida are members of the EuroFlow Consortiu

Ichthyofauna Used in Traditional Medicine in Brazil

Fish represent the group of vertebrates with the largest number of species and the largest geographic distribution; they are also used in different ways by modern civilizations. The goal of this study was to compile the current knowledge on the use of ichthyofauna in zootherapeutic practices in Brazil, including ecological and conservational commentary on the species recorded. We recorded a total of 85 species (44 fresh-water species and 41 salt-water species) used for medicinal purposes in Brazil. The three most commonly cited species were Hoplias malabaricus, Hippocampus reidi, and Electrophorus electricus. In terms of conservation status, 65% of species are in the “not evaluated” category, and 14% are in the “insufficient data” category. Three species are in the “vulnerable” category: Atlantoraja cyclophora, Balistes vetula, and Hippocampus erectus. Currently, we cannot avoid considering human pressure on the population dynamics of these species, which is an essential variable for the conservation of the species and the ecosystems in which they live and for the perpetuation of traditional medical practices

The contribution of meteorological parameters and the COVID-19 partial lockdown on air quality in Rio de Janeiro, Brazil

This study evaluated the pollutant levels (NO2, SO2, CO, and O3), air quality index (AQI) and the influence of meteorological variables and coronavirus disease (COVID-19) pandemic on the air quality in Rio de Janeiro. The data set used comprises periods before (March-April, 2019) and during pandemic (March-April, 2020). According to the AQI results, on most days, the air quality was ranked as “good”. Brazilian air quality standards for SO2, O3, and NO2 were not exceeded in any of the monitoring stations during partial lockdown, while CO exceeded in all periods in one site due to industrial emission. Comparing both periods, descriptive statistics for the meteorological parameters presented no differences, which suggests similar conditions. However, when evaluated week by week in 2020, weather conditions presented some differences that probably affected pollutant concentrations. The correlations between O3 and NO2 and some meteorological parameters indicate that variations in both favored ozone formation, since it is a photochemical process favored by temperature and solar radiation and that, in Rio de Janeiro, low NO2 concentrations lead to increased O3. The improvements on air quality during the partial lockdown may be attributed mainly to a reduction on emission sources rather than weather conditions.Campus Lima Centr