Promoter regions of Plasmodium vivax are poorly or not recognized by Plasmodium falciparum

BACKGROUND: Heterologous promoter analysis in Plasmodium has revealed the existence of conserved cis regulatory elements as promoters from different species can drive expression of reporter genes in heterologous transfection assays. Here, the functional characterization of different Plasmodium vivax promoters in Plasmodium falciparum using luciferase as the reporter gene is presented. METHODS: Luciferase reporter plasmids harboring the upstream regions of the msp1, dhfr, and vir3 genes as well as the full-length intergenic regions of the vir23/24 and ef-1α genes of P. vivax were constructed and transiently transfected in P. falciparum. RESULTS: Only the constructs with the full-length intergenic regions of the vir23/24 and ef-1α genes were recognized by the P. falciparum transcription machinery albeit to values approximately two orders of magnitude lower than those reported by luc plasmids harbouring promoter regions from P. falciparum and Plasmodium berghei. A bioinformatics approach allowed the identification of a motif (GCATAT) in the ef-1α intergenic region that is conserved in five Plasmodium species but is degenerate (GCANAN) in P. vivax. Mutations of this motif in the P. berghei ef-1α promoter region decreased reporter expression indicating it is active in gene expression in Plasmodium. CONCLUSION: Together, this data indicates that promoter regions of P. vivax are poorly or not recognized by the P. falciparum transcription machinery suggesting the existence of P. vivax-specific transcription regulatory elements

A Non Mouse-Adapted Dengue Virus Strain as a New Model of Severe Dengue Infection in AG129 Mice

The spread of dengue (DEN) worldwide combined with an increased severity of the DEN-associated clinical outcomes have made this mosquito-borne virus of great global public health importance. Progress in understanding DEN pathogenesis and in developing effective treatments has been hampered by the lack of a suitable small animal model. Most of the DEN clinical isolates and cell culture-passaged DEN virus strains reported so far require either host adaptation, inoculation with a high dose and/or intravenous administration to elicit a virulent phenotype in mice which results, at best, in a productive infection with no, few, or irrelevant disease manifestations, and with mice dying within few days at the peak of viremia. Here we describe a non-mouse-adapted DEN2 virus strain (D2Y98P) that is highly infectious in AG129 mice (lacking interferon-α/β and -γ receptors) upon intraperitoneal administration. Infection with a high dose of D2Y98P induced cytokine storm, massive organ damage, and severe vascular leakage, leading to haemorrhage and rapid death of the animals at the peak of viremia. In contrast, very interestingly and uniquely, infection with a low dose of D2Y98P led to asymptomatic viral dissemination and replication in relevant organs, followed by non-paralytic death of the animals few days after virus clearance, similar to the disease kinetic in humans. Spleen damage, liver dysfunction and increased vascular permeability, but no haemorrhage, were observed in moribund animals, suggesting intact vascular integrity, a cardinal feature in DEN shock syndrome. Infection with D2Y98P thus offers the opportunity to further decipher some of the aspects of dengue pathogenesis and provides a new platform for drug and vaccine testing

Carbon Dioxide Utilisation -The Formate Route

UIDB/50006/2020 CEEC-Individual 2017 Program Contract.The relentless rise of atmospheric CO2 is causing large and unpredictable impacts on the Earth climate, due to the CO2 significant greenhouse effect, besides being responsible for the ocean acidification, with consequent huge impacts in our daily lives and in all forms of life. To stop spiral of destruction, we must actively reduce the CO2 emissions and develop new and more efficient “CO2 sinks”. We should be focused on the opportunities provided by exploiting this novel and huge carbon feedstock to produce de novo fuels and added-value compounds. The conversion of CO2 into formate offers key advantages for carbon recycling, and formate dehydrogenase (FDH) enzymes are at the centre of intense research, due to the “green” advantages the bioconversion can offer, namely substrate and product selectivity and specificity, in reactions run at ambient temperature and pressure and neutral pH. In this chapter, we describe the remarkable recent progress towards efficient and selective FDH-catalysed CO2 reduction to formate. We focus on the enzymes, discussing their structure and mechanism of action. Selected promising studies and successful proof of concepts of FDH-dependent CO2 reduction to formate and beyond are discussed, to highlight the power of FDHs and the challenges this CO2 bioconversion still faces.publishersversionpublishe

Armazenamento de frutos de quiabo embalados com filme de PVC em condição ambiente Shelf life of four cultivars of okra covered with PVC film at room temperature

O presente trabalho objetivou avaliar a influência do filme de PVC durante o armazenamento, em condição ambiente, de frutos de quatro cultivares de quiabo para consumo in natura. O experimento foi organizado segundo delineamento experimental em blocos casualizados, em esquema de parcelas subdivididas, tendo nas parcelas um fatorial 2x4, ou seja, embalagens sem e com PVC e quatro cultivares (Amarelinho, Red Velvet, Star of David, Mammoth Spinless), e nas subparcelas os seis períodos de amostragem com quatro blocos. As características avaliadas foram: massa fresca, teor relativo de água, teor de vitamina C, teor de clorofila total e análise visual do escurecimento. O uso do PVC proporcionou menor perda de massa fresca e maior teor relativo de água ao longo do armazenamento para todas as cultivares estudadas em relação às não embaladas com o filme. As menores perdas de massa fresca foram verificadas nas cvs. Amarelinho e Star of David sem e com PVC, respectivamente. Os frutos das cvs. Red Velvet e Star of David, embalados com PVC, apresentaram menores perdas no teor de vitamina C. Foi observado que a cv. Mammoth Spinless manteve maior teor de clorofila. Observou-se, de maneira geral, maior incidência de escurecimento nos frutos armazenados sem PVC. Verificou-se que de toda as cultivares estudadas o que manifestou melhor conservação com relação ao escurecimento foi o Red Velvet.<br>The present work evaluated the influence of PVC film on the postharvest shelf life of four cultivars of okra stored at room temperature. The experiment was arranged in random blocks, in sub split parcels, where the parcels were a factorial 2x4, with and without PVC film and the cvs. Amarelinho, Red Velvet, Star of David and Mammoth Spinless, and the subparcels the six sample time with four blocks. The characteristics evaluated were: percentage of weight loss, relative water content, total chlorophyll and chlorophyll a and b, content of vitamin C, visual browning. The PVC was efficient in controlling the weight loss and retained the water content through storage. The lowest losses were observed for the cvs. Amarelinho and Star of David, without and with PVC, respectively. There was higher retention of vitamin C in the fruits wrapped with PVC and the cvs. Red Velvet and Star of David kept higher content of it. In general, the cv. Mammoth Spinless kept higher content of chlorophyll. The higher incidence of browning was in fruits stored without PVC. The Red Velvet had better shelf life due to lower browning

Appraisal of surgical treatment of 47 cases of patellofemoral instability Avaliação do tratamento cirúrgico da instabilidade fêmoro-patelar em 47 casos

INTRODUCTION: Patellofemoral instability is a common knee disease. Its etiology is complex and variable, with many components making different contributions in each individual, resulting in several distinct clinical presentations. Our goal was to analyze the results of surgical treatment in our hospital over a period of 10 years. PATIENTS AND METHODS: We analyzed 55 knees of 47 patients who underwent surgery for patellofemoral instability and were classified into 2 main groups: proximal realignment and combined proximal and distal realignment. Three other groups were analyzed according to the duration of preoperative symptoms: less than 1 year (group I); 1 to 10 years (group II); and more than 10 years (group III). RESULTS: There were 62% good results overall, with 78% good results in groups I and II. Group III had 81% bad results, showing that a late diagnosis of advanced disease results in a poor prognosis. In addition to late diagnosis, bad results were usually associated with incorrect diagnosis or choice of surgical technique. There was no significant difference between isolated proximal realignment and combined proximal and distal realignment in groups I or II, but in group III, the combined technique yielded better results. DISCUSSION: Our results indicate that patellofemoral instability should be addressed in its early stages. Patients with long-lasting symptoms or more severe disease seem to achieve better results with combined techniques. CONCLUSION: Proximal and distal realignments produce better results than isolated proximal realignment in patients with joint degeneration or with greater duration of disease. The realignment surgery does not produce good results in patients with advanced disease.<br>A instabilidade fêmoro-patelar (IFP) é patologia freqüente cuja etiologia é complexa e variável com diversos componentes cuja importância varia em cada indivíduo, resultando em diversas apresentações clínicas. Nosso objetivo foi analisar os resultados do tratamento cirúrgico em um período de 10 anos. MATERIAL E MÉTODO: Nós analisamos 55 joelhos de 47 pacientes operados por IFP em dois grupos principais: realinhamento proximal e realinhamento proximal e distal. Três outros grupos de acordo com a duração dos sintomas: <1 ano (grupo I); entre 1 e 10 (grupo II); e >10 anos (grupo III). RESULTADOS: Obtivemos 62% de bons resultados globalmente e 78% de bons resultados nos grupos I e II. O grupo III apresentou 81% de maus resultados, demonstrando que a indicação tardia não é boa. Os maus resultados, além do já mencionado, estavam em geral associados a erros diagnósticos ou de escolha de técnica cirúrgica. Não houveram diferenças significantes entre os resultados do realinhamento proximal isolado e do realinhamento proximal e distal combinados nos grupos I e II mas no grupo III o realinhamento combinado apresentou melhores resultados. DISCUSSÃO: Nossos resultados indicam que a instabilidade fêmoro-patelar deve ser tratada na fase inicial. Nos casos de longa evolução e nos casos mais graves, o realinhamento proximal e distal tem melhores resultados. CONCLUSÃO: O realinhamento proximal e distal tem melhor resultado que o realinhamento proximal isolado em pacientes com alterações degenerativas e naqueles com longa evolução. A operação de realinhamento não apresenta bons resultados nos casos de doença avançada