Expression of anti-Z-DNA single chain antibody variable fragment on the filamentous phage surface

We describe the expression of an anti-Z-DNA single chain variable region antibody fragment (scFv) on a filamentous phage surface. Four vectors for phage display were constructed. Two of them are able to display multiple copies of the antibody fragment, and the others can be used to make monovalent libraries. The vectors use different promoter/leader sequences to direct the expression of the fused proteins. All were able to promote the assembly of fusion virion particles. In this paper we also show the affinity selection (biopanning) of those phage-antibodies based on the capacity of their products to recognize the antigen. We used biotinylated Z-DNA and the selection was performed in a solution phase fashion. The data presented here indicate that these vectors can be further used to construct anti-nucleic acid antibody fragment libraries that can be used to study the basis of nucleic acid-protein interaction and its role in autoimmunity mechanisms

Optimization of heterologous protein production in Chinese hamster ovary cells under overexpression of spliced form of human X-box binding protein

Background: The optimization of protein production is a complex and challenging problem in biotechnology. Different techniques for transcription, translation engineering and the optimization of cell culture conditions have been used to improve protein secretion, but there remain many open problems involving post-translational modifications of the secreted protein and cell line stability. Results: In this work, we focus on the regulation of secreted protein specific productivity (using a recombinant human immunoglobulin G (IgG)) by controlling the expression of the spliced form of human X-box binding protein (XBP-(s)) in Chinese hamster ovary cells (CHO-K1) under doxycycline (DOX) induction at different temperatures. We observed a four-fold increase in specific IgG productivity by CHO cells under elevated concentrations of DOX at 30°C compared to 37°C, without detectable differences in binding activity in vitro or changes in the structural integrity of IgG. In addition, we found a correlation between the overexpression of human XBP-1(s) (and, as a consequence, endoplasmic reticulum (ER) size expansion) and the specific IgG productivity under DOX induction. Conclusions: Our data suggest the T-REx system overexpressing human XBP-1(s) can be successfully used in CHO-K1 cells for human immunoglobulin production

Uma abordagem diacrônica sobre a influência da relação Brasil-Estados Unidos no mapeamento do território brasileiro nas escalas topográficas 1:50.000 e 1:100.000

Uma das características do território brasileiro é sua dimensão continental. Até a década de 1960, esse vasto território (8.514.876 Km²) foi mapeado em sua integridade somente na escala de 1:1.000.000. O Brasil possuía nessa época apenas algumas insignificantes ilhas de mapeamento em escalas maiores, como as escalas topográficas de 1:100.000 e 1:50.000 (respectivamente, 12,8% e 5,7% do território). A representação cartográfica nessas escalas era uma necessidade, mas só viria a ser iniciada em contextos políticos favoráveis, como o da aproximação política dos EUA com Brasil na década de 1950, visando garantir a defesa do hemisfério ocidental. Essa mesma política é reforçada na década seguinte, no contexto político da Guerra Fria e de programas especiais para a América Latina como a “Aliança para o Progresso”, proposta no governo John F. Kennedy. Neste âmbito, é posto em ação um extenso aerolevantamento fotogramétrico do território brasileiro, executado pelo 10º Esquadrão de Levantamento Aéreo do Exército Norte Americano (Aerial Squadron Team 10º - USA).One of the characteristics of the Brazilian territory is your continental dimension. Until the 1960s, this large territory (8,514,876 km²) was mapped only on the scale of 1:1.000.000. At that time, Brazil had only a few insignificant islands of mapping in larger scales, such as the 1:100.000 and 1:50.000 topographic scales (12,8% and 5,7% of the territory respectively). Cartographic representation on these scales was a necessity, but it would only be initiated in favorable political contexts, such as that of the US political rapprochement with Brazil in the 1950s, in order to guarantee the defense of the Western. This same policy is reinforced in the following decade, in the context of the Cold War and special programs for Latin America such as “The Alliance for Progress" proposed by the John F. Kennedy administration. In this context, an extensive photogrammetric aero-survey of the Brazilian territory is carried out, executed by the 10th Aerial Squadron of the North American Army.Una de las características del territorio brasileño es su dimensión continental. Hasta la década de 1960, ese vasto territorio (8.514.876 Km²) fue mapeado en su integridad solamente en la escala de 1: 1.000.000. En esa época, Brasil contaba con apenas algunas zonas pequeñas y aisladas que resultaban insignificantes dentro del cuadro general y sólo en escalas mayores, tales como 1: 100.000 y 1: 50.000 (el 12,8% y el 5,7% del territorio, respectivamente). La representación cartográfica en esas escalas era una necesidad para la gestión estatal, pero sólo fue posible y se inició cuando se configuraron contextos políticos favorables, como el de la aproximación política de Estados Unidos con Brasil en la década de 1950 (originalmente promovido con el fin de garantizar la defensa del hemisferio occidental). Esta misma política se reforzó en la década siguiente, en el contexto político de la Guerra Fría y de la implementación de programas especiales para América Latina como la "Alianza para el Progreso" (propuesta en el gobierno de John F. Kennedy). En este ámbito, se puso en acción un extenso relevamiento fotogramétrico del territorio brasileño, ejecutado por el 10º Escuadrón de Levantamiento Aéreo del Ejército Norteamericano (Aerial Squadron Team 10º - USA).Une des caractéristiques du territoire brésilien est sa dimension continentale. Jusqu’aux années 1960, ce vaste territoire (8.514.876 km²) n'était entièrement cartographié que sur une échelle de 1: 1.000.000. À l'époque, le Brésil avait des cartes de quelques petites zones isolées qui étaient insignifiants dans le cadre général et seulement dans des échelles plus anciennes comme 1: 100.000 et 1: 50 000 (12,8% et 5,7% du territoire, respectivement). La représentation cartographique à ces échelles est nécessaire pour la gestion de l'État, mais la production de ces cartes a devenue possible et a commencée lorsque des contextes politiques favorables, comme l'approche politique des États-Unis avec le Brésil dans les années 1950 (à l'origine, pour garantir la défense de l'hémisphère occidental). Cette même politique a été renforcée dans la prochaine décennie, dans le contexte politique de la guerre froide et la mise en œuvre de programmes spéciaux pour l'Amérique latine comme « Alliance pour le progrès » (proposé dans le gouvernement de John F. Kennedy). Dans cette zone, un vaste projet photogrammétrique du territoire brésilien a exécutée par la 10ème escadrille aérienne américaine (Aerial Squadron Team 10º - USA)

The efficacy of humanized antibody against the Sporothrix antigen, gp70, in promoting phagocytosis and reducing disease burden

Sporotrichosis is a subcutaneous mycosis distributed worldwide and is frequently reported in countries with tropical climates, as Latin America countries. We previously demonstrated that mice with sporotrichosis produce specific antibodies against a 70-kDa fungal protein, indicating that specific antibodies against this molecule may help to control the sporotrichosis. IgG1 monoclonal antibody was generated, and called mAbP6E7, in mice against a 70-kDa glycoprotein (gp70) of S. schenckii. The mAbP6E7 showed prophylactic and therapeutic activity against sporotrichosis. However, this antibody has a murine origin, and this can generate an immune response when administered to humans, precluding its use for a prolonged time. For its possible use in the treatment of human sporotrichosis, we humanized the mAbP6E7 by genetic engineering. Once expressed, the humanized antibodies had good stability and were able to bind to the 70-kDa cell wall antigens of Sporothrix schenckii and S. brasiliensis. The humanized P6E7 were able to opsonize S. schenckii yeasts, thus increasing the phagocytic index in human monocyte-derived macrophages. The treatment with humanized P6E7 decreased fungal burden in vivo. These data suggest that humanized P6E7 may have a therapeutic role in sporotrichosis

MicroRNA expression profiles in human CD3+ T cells following stimulation with anti‑human CD3 antibodies

Background: Anti-CD3 therapy can induce immunosuppression by several non mutually exclusive mechanisms that have been proposed to explain the therapeutic effect the administration anti-CD3 mAb, but its immunoregulatory mechanism is still not completely clear. In T cells, microRNAs (miRNAs) regulate several pathways, including those associated with immune tolerance. Here, we report changes in miRNA expression in T cells following treatment with anti-human CD3 antibodies. Peripheral blood mononuclear cells were cultured in the presence of the monoclonal antibody OKT3 or a recombinant fragment of humanized anti-CD3. Following these treatments, the expression profiles of 31 miRNA species were assessed in T cells using TaqMan arrays. Results: Eight of the tested miRNAs (miR-155, miR-21, miR-146a, miR-210, miR-17, miR-590-5p, miR-106b and miR- 301a) were statistically significantly up- or down-regulated relative to untreated cells. Conclusions: Stimulation of T cells with anti-human CD3 antibodies alters miRNA expression patterns, including of miRNA species associated with immune regulatory pathways

O canto de soltura de Rhinella crucifer (Wied-Neuwied, 1821), no Estado de Pernambuco, Nordeste do Brasil

In this work, we describe for the first time the release call of Rhinella crucifer in addition to comparing its advertisement call with other species of your group. The records were made in an area of Atlantic Forest, named Refúgio de Vida Silvestre Matas de Sirij, in the municipality of São Vicente Férrer, state of Pernambuco, in the Northeast Region of Brazil. At 18 June 2019, 24 advertisement vocalizations of four males were recorded, of which three emit 31 release calls. The results showed a structural difference between the two types of vocalizations, such as the presence of pulses defined, harmonics and greater modulation in the release call. Compared to other Rhinella species of the crucifer group, the release call of R. crucifer had a greater resemblance to R. ornata in the duration and interval of notes and number and interval of pulses. While the proximity of the dynamic parameters of the advertisement call with R. casconi, the highlight was the compatibility of the dominant frequency with R. inopina. This work adds new information on the vocal repertoire of the species, which contributes to information about your natural history.Neste trabalho descrevemos pela primeira vez o canto de soltura de Rhinella crucifer além de comparar o seu canto de anúncio com outras espécies de seu grupo. Os registros foram feitos em uma área de Floresta Atlântica denominada Refúgio de Vida Silvestres Matas de Sirij, no município de São Vicente Férrer, Pernambuco, na Região nordeste do Brasil. No dia 18 de junho de 2019 foram registradas 24 vocalizações de anúncio de quatro machos, dos quais três reproduziram 31 chamadas de soltura. Os resultados mostraram que houve diferença estrutural entre os dois tipos de cantos, como a presença de pulsos definidos, harmônicos e maior modulação no canto de soltura. Quando comparado com outras espécies de Rhinella do grupo crucifer, o canto de soltura do R. crucifer teve maior semelhança com R. ornata na duração e intervalo de notas e número e intervalo de pulsos. Apesar da proximidade dos parâmetros dinâmicos do canto de anuncio com R. casconi, o destaque foi a similaridade da frequência dominante com R. inopina. Com este trabalho adicionamos mais informações do repertório vocal da espécie, o que contribui com informações sobre sua história natural.Asociación Herpetológica Argentin

Violacein Extracted from Chromobacterium violaceum Inhibits Plasmodium Growth in Vitro and in Vivo

Violacein is a violet pigment extracted from the gram-negative bacterium Chromobacterium violaceum. It presents bactericidal, tumoricidal, trypanocidal, and antileishmanial activities. We show that micromolar concentrations efficiently killed chloroquine-sensitive and -resistant Plasmodium falciparum strains in vitro; inhibited parasitemia in vivo, even after parasite establishment; and protected Plasmodium chabaudi chabaudi-infected mice from a lethal challenge.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Estadual Campinas, UNICAMP, Dept Parasitol, Inst Biol, BR-13083970 Campinas, SP, BrazilUniv Estadual Campinas, UNICAMP, Dept Microbiol & Imunol, Inst Biol, BR-13083970 Campinas, SP, BrazilUniversidade Federal de São Paulo, UNIFESP, Dept Bioquim, BR-04044020 São Paulo, BrazilCEPEM, IPEPATRO, BR-78900970 Porto Velho, RO, BrazilUniv São Paulo, Dept Parasitol, ICB2, São Paulo, BrazilUniv Estadual Campinas, Dept Fisiol & Biofis, Inst Biol, BR-13083970 Campinas, SP, BrazilUniv Estadual Campinas, Lab Quim Biol, Inst Quim, BR-13083970 Campinas, SP, BrazilUniversidade Federal de São Paulo, UNIFESP, Dept Bioquim, BR-04044020 São Paulo, BrazilFAPESP: 2004/00638-6CNPq: 470587/2006-7Web of Scienc

Inflammation and circulating endothelial progenitor cells in patients with coronary artery disease and residual platelet reactivity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Albert Einstein HospitalUniversidade de São Paulo Faculdade de Medicina Hospital das ClínicasDuke University Medical Center Duke Clinical Research InstituteUniversidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM)UNIFESP, EPMFAPESP: 05/57710-3SciEL

DETERIORAÇÃO DE ÓLEOS VEGETAIS EXPOSTOS A DIFERENTES CONDIÇÕES DE ARMAZENAMENTO

The final quality of the oil depends on all the stages of the production. These steps range from the cultivation, extraction, refining, transportation, distribution and marketing of oils. Oxidation processes in foods are responsible for reducing shelf life of raw materials and manufactured products leading to major economic loss. The objective of this study was to analyze the levels of acidity and peroxide virgin vegetable oils from soy, corn, canola and sunflower undergo under different conditions of storage before consumption. The procedure for checking the index of acidity and peroxide was conducted based on the methodology of the Adolfo Lutz Institute (2004). At the end of the analysis period, all oil exposed group had higher acid value and peroxide to the saved group. Soybean oil was found to be the most stable oil to work in relation to the indices evaluated, followed by sunflower oil. Despite the canola oil have low levels of saturated fatty acids, this research showed that canola oil is an extremely sensitive regarding their physicochemical stability when exposed to light and temperature.A qualidade final de um óleo depende de todas as etapas envolvidas na produção do mesmo. Estas etapas incluem desde o cultivo, a extração, o refino, o transporte, a distribuição e a comercialização dos óleos. Os processos de oxidação em alimentos são responsáveis pela redução da vida na prateleira das matérias-primas e dos produtos industrializados levando a importante perda econômica. O objetivo deste trabalho foi analisar os índices de acidez e peróxido de óleos vegetais virgens de soja, milho, canola e girassol, submetidos a diferentes condições de armazenamento antes do consumo. O procedimento para verificação do índice de acidez e peróxido foi executado com base na metodologia do Instituto Adolf Lutz (2004). Ao final do período de análise, todos os óleos do grupo exposto apresentavam índice de acidez e peróxido superiores ao do grupo guardado. O óleo de soja mostrou-se o óleo mais estável para se trabalhar em relação aos índices avaliados, seguido do óleo de girassol. Apesar do óleo de canola apresentar baixos teores de ácidos graxos saturados, esta pesquisa revelou que o canola é um óleo extremamente sensível em relação a sua estabilidade físico- química quando exposto a luz e a temperatura

SISTEMA DE ANÁLISE ESTEQUIOMÉTRICA PARA PRODUÇÃO DE SABÃO A PARTIR ÓLEO VEGETAL RESIDUAL: UMA ESTRATÉGIA PARA REDUÇÃO DO IMPACTO AMBIENTAL

The manufacture of soap using waste vegetable oil is one of the ways to avoid it reaches water bodies and soil in its ruling, thus mitigating their impacts on the environment. His artisanal does not require much investment and knowledge, it is a simple process that uses inputs and equipment used in the daily lives of most people. This work aims at presenting the results of an analysis system that would provide a good reason between the inputs used in the production of soap from vegetable oil waste. Based on these results, it was made possible the production of a soap with acceptable physical and chemical characteristics, of satisfactory quality similar to industrialized and, indirectly, the awareness of the inappropriate disposal of vegetable oil.A fabricação de sabão utilizando óleo vegetal residual é uma das formas de evitarque este alcance os corpos hídricos e o solo na sua forma prejudicial,mitigando assim seus impactos ao meio ambiente. Sua fabricação artesanal nãorequer muitos investimentos e conhecimentos, pois é um processo simples queutiliza insumos e equipamentos utilizados no cotidiano da maioria das pessoas.Este trabalho teve por objetivo apresentar os resultados de um sistema deanálise que proporcionasse uma razão ótima entre os insumos utilizados naprodução de sabão a partir do óleo vegetal residual. Baseado nestes resultados,foi viabilizada a produção de um sabão com características físicas e químicasaceitáveis, de qualidade satisfatória semelhantes aos industrializados e, indiretamente,a conscientização quanto ao descarte inadequado do óleo vegetal