Cinetic Study of acute episode of bacterial translocation and it’s immunological and microcirculatory repercussion in rats

Introdução: Muitos pacientes ainda morrem devido a infecções. Crescentes relatos da literatura atual têm atribuído ao intestino o papel de agravamento de doenças graves, e/ou a sua participação na gênese da sepse por mecanismo de translocação bacteriana (TB). Objetivo: Avaliar de forma cinética a TB e suas repercussões microcirculatórias e imunológicas. Método: Ratos Wistar-EPM (n=162) foram aleatoriamente distribuídos em grupo Sham (n=72) e grupo TB (n=90), e avaliados nos períodos de 2h, 6h, 24h, 72h, 7 e 14 dias, em relação a índice de translocação bacteriana; microscopia intravital; perfusão tecidual; e componentes celulares e humorais da linfa mesentérica por linfograma, citometria de fluxo e CBA-Flex. Resultado: A TB ocorreu somente no grupo TB e foi expressiva nas primeiras 24 horas tornando-se negativa somente com 7 dias. A conseqüência de um episódio de TB repercutiu na celularidade e citocinas pró e antiinflamatórias da linfa mesentérica associado a lesões da microcirculação e hipoperfusão tecidual de forma local e sistêmica. A citometria de fluxo mostrou que a linfa mesentérica eferente pós TB difere significativamente do grupo Sham quanto a número e subpopulação de linfócitos. Conclusão: Um episódio agudo de TB determinou uma recuperação máxima bacteriana com 6 horas e sua completa depuração entre 3 e 7 dias, além de provocar alterações da microcirculação intestinal e sistêmica associadas à ativação do GALT, principalmente no período de permanência das bactérias translocadas no hospedeiro, sendo a via linfática mesenterial uma importante rota na intercomunicação imunológica entre o ambiente intestinal e sistêmico.Introduction: Many patients still die from infection. Currently, growing evidences have pointed out the role of the gut in the worsening of the critical illness, and/or its participation in the genesis of the bacterial translocation. Objective: Evaluate the bacterial translocation (BT) kinetics and its repercussion on microcirculation and immune response. Method: Wistar-EPM rats (n=162) were randomly distributed in Sham group (n=72) and BT group (n=90), and were monitored at 2h, 6h, 24h, 72h, 7 and 14 days periods in relation to bacterial translocation index, intravital microscopy, tissue perfusion index, and cellular and humoral components of the mesenteric lymph by lymphogram, flow cytometry and CBA-Flex. Results: Bacterial recovery was positive only in BT-group and it was expressive in the first 24 hours, becoming negative only after seven days. The consequences of one episode of BT could be seen in the cellularity and proinflammatory and antiinflammatory citokines of the efferent mesenteric lymph associated to local and systemic microcirculation and tissue hypoperfusion. The flow cytometry showed that efferent mesenteric lymph after BT was significantly different as compared to Sham group in relation to lymphocites number and their subtype. Conclusion: An acute episode of BT determined maximal bacterial recovery at 6h and its complete clearance occurred between 3 and 7 days, in addition to the gut and systemic microcirculation injuries due to the GALT activation, specially at the presence of translocated bacteria in the host, demonstrating the importance of the lymphatic route in the immune crosstalk between the gut and systemic enviroment.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP: 05/53826-

Aplicações multilineares completamente absolutamente somantes

Orientador : Mario Carvalho de MatosTese (doutorado) - Universidade Estadual de Campinas, Instituto de Matematica, Estatistica e Computação CientificaDoutoradoDoutor em Matemátic

Preparação e escalonamento de um hidrolisado enzimático de proteínas do soro de leite bovino

Foi descrita a preparação de um hidrolisado de proteínas do soro de leite bovino com enzimas pancreáticas, contendo baixos níveis de aminoácidos aromáticos. Quando utilizadas pancreatina e tripsina/quimotripsina, por 27h a 37±2ºC, foram liberados 74% de Phe, 100% de Tyr e 100% de Trp como aminoácidos livres. A maioria dos aminoácidos aromáticos livres, presentes em dois quilos de hidrolisado (15 litros), foi separada dos peptídeos e outros aminoácidos remanescentes por filtração em coluna de gel de Sephadex G-25C eluída com ácido acético 5%, fluxo de 60 litros por hora a 25ºC. O produto, recuperado com 37% de rendimento, continha 0,70 mmol de Phe, 0,41 mmol de Tyr eWe describe the preparation of pancreatic enzymes hydrolysate of milk whey proteins containing low levels of aromatic amino acids. Pancreatin and trypsin/chymotrypsin (6.3% w/w protein) when used to hydrolyze whey proteins for 27 h at 37±2 ºC, released 74% of the Phe, 100% of the Tyr and 100% of the Trp as free amino acids. Most of the free aromatic amino acids present in 2 kg hydrolysate were separated from the remaining peptides and other amino acids by gel filtration on a 15 liter Sephadex G-25 column eluted with 5% acetic acid at 60 liters h-1 at 25ºC. The product, recovered in 37% yield, contained 0.70 mmol Phe, 0.41 mmol Tyr, an

Functionalized Boron Nitride Applications in Biotechnology

Due to its interesting chemical, physical, and biological properties, boron nitride has received considerable attention by the scientific and technological communities. However, there is a strong dependency of its structural quality and compatibility in different host systems, regarding its potential applications. The use of these different nanostructures involves several challenges due to their low dispersibility in water and organic solvents; thus, its chemical modification is an important step that gives them specificity. Therefore, the ability to control their surface (physically or chemically) is essential for exploring and building blocks in the nanoengineering of supramolecular structures. In this chapter, we report different boron nitride functionalization processes, as well as their important uses as adjuvants in vaccines, brachytherapy, or drug delivery. Besides some important theoretical studies that have demonstrated the different functionalization possibilities for use in nanomedicine, are also reported

Protective effect and expression of defense-related ESTs induced by acibenzolar-S-methyl and a phosphorylated mannan oligosaccharide-based product against Moniliophthora perniciosa in Theobroma cacao

Witches’ broom disease (WBD), caused by the fungus Moniliophthora perniciosa, is one of the main diseases in cocoa (Theobroma cacao) and has caused severe economic losses. Integrated disease management has been the focus for its control and therefore, the identification of new inducers of plant resistance is desirable. Thus, the goal of this work was to evaluate two potential inducers of resistance against WBD. A phosphorylated mannan oligosaccharide-based product (PMO) and acibenzolar S-methyl (ASM) were tested on M. perniciosa inoculated seedlings and in field experiments and showed a reduction on the incidence of WBD. The expression of two defense-related expressed sequence tags (ESTs) in cocoa, coding for peroxidase (Pox) and chitinase (Chi), were accessed by qPCR. Both products induced the expression of the Pox defense-related EST. In general, ASM induced the expression of chitinase (Chi) and peroxidase (Pox) in earlier time-points than PMO. However, PMO provided long-lasting and higher levels of expression. Chi expression was triggered in the time-points succeeding the spraying but was very low. On the other hand, peaks of Pox transcripts were detected in later time-points for both inducers. ASM and PMO modes of action might be explained, at least partially, by the overexpression of defense-related ESTs.Keywords: Cocoa, witches’ broom disease, disease control, peroxidase, chitinase, induced resistance, elicitors, quantitative reverse transcriptase PCR (qRT-PCR)African Journal of Biotechnology Vol. 12(12), pp. 1311-131

Eficiência de protocolos de extração de RNA em diferentes tecidos do cafeeiro

In order to use sensitive techniques of molecular biology, such as the study of differentially expressed genes, a highqualityRNA in suitable quantities is necessary. Due to the presence of several varieties and often expressive quantities of secondarycompounds in plants, there is no standard method for the isolation of nucleic acids that can be used for all species. Polyphenols andpolysaccharides are the compounds that interfere the most in the extraction process, and when they are present, a low-quality RNAis produced. Four RNA extraction methods (CTAB method, Hot Borate, CONCERT and Tri Reagent), in four different coffee tissues(root, leaf, flower and fruit) were tested in this work, aiming at determining which method is more efficient. It was observed that theCTAB and Hot Borate methods, in which PVP and/or -mercaptoethanol were added and precipitation with LiCl was performed,presented more pure RNA, with no degradation observed in any of the tissues, being suitable for further gene expression analysis.High-quality RNA was not obtained from any tissue in the extraction with Tri Reagent, which includes the use of phenol, and thusexpression analysis was disturbed. The CTAB macroextraction method presented samples with the highest RNA quality and largestquantities in all tissues. Future works need to be carried out aiming the standardization of this macroextraction method.Para a utilização de técnicas sensíveis de biologia molecular, como o estudo de genes diferencialmente expressos, énecessário a obtenção de um RNA de boa qualidade e em quantidades adequadas. Devido à presença de grandes variedades, efrequentemente grande quantidade de compostos secundários em plantas, não existe um método padrão para o isolamento de ácidosnucléicos que possa ser utilizado para todas as espécies. Os polifenóis e os polissacarídeos são os compostos de maior interferênciano processo de extração, e quando presentes geram um RNA de baixa qualidade. Nesse trabalho foram testados quatro métodos deextração de RNA (Método CTAB, Borato quente, CONCERT e Tri Reagente), em quatro diferentes tecidos de café (raiz, folha, flor efruto), objetivando-se determinar qual método é mais eficiente. Foi observado que os métodos, CTAB e Borato quente, que possuíama adição PVP e/ou -mercaptoetanol, e precipitação com LiCl, foram os que apresentaram RNAs mais puros e sem degradação emtodos os tecidos, e puderam ser utilizados para a análise de expressão gênica. Com a extração utilizando o TriReagente, que tem comobase o fenol, não foi obtido RNA de boa qualidade em todos os tecidos e consequentemente não foi possível a análise de expressão. Ométodo de macroextração CTAB foi o que apresentou amostras com RNA de melhor qualidade e em grandes quantidades em todosos tecidos. Trabalhos posteriores precisam ser realizados a fim de padronizar esse método para microextração

Immunotherapy in patients with advanced non-small cell lung cancer lacking driver mutations and future perspectives

From a complete literature review, we were able to present in this paper what is most current in the treatment with immunotherapy for advanced non-small cell lung cancer (NSCLC). Especially the use of immunotherapy, particularly inhibitors of PD-1 (programmed cell death protein 1), PDL-1 (programmed cell death protein ligand 1), and CTLA-4 (cytotoxic T-lymphocyte antigen 4). Since 2015, these drugs have transformed the treatment of advanced NSCLC lacking driver mutations, evolving from second-line therapy to first-line, with excellent results. The arrival of new checkpoint inhibitors such as cemiplimab and the use of checkpoint inhibitors earlier in the therapy of advanced and metastatic cancers has been making the future prospects for treating NSCLC lacking driver mutations more favorable and optimistic. In addition, for those patients who have low PDL-1 positivity tumors, the combination of cytotoxic chemotherapy, VEGF inhibitor, and immunotherapy have shown an important improvement in global survival and progression free survival regardless the PDL-1 status. We also explored the effectiveness of adding radiotherapy to immunotherapy and the most current results about this combination. One concern that cannot be overlooked is the safety profile of immune checkpoint inhibitors (ICI) and the most common toxicities are described throughout this paper as well as tumor resistance to ICI.info:eu-repo/semantics/publishedVersio

Resistência de genótipos de tomateiro com alto teor de acilaçúcares a Tetranychus evansi Baker & Pritchard

O acesso de Lycopersicon pennellii LA716 tem sido utilizado em programas de melhoramento do tomateiro devido à sua resistência a artrópodos-pragas, mediada pela presença de acilaçúcares nos folíolos. Foi verificada a correlação entre o teor de acilaçúcares em folíolos de tomateiros e a repelência ao ácaro Tetranychus evansi Baker & Pritchard. Foram selecionadas plantas contrastantes quanto aos níveis de acilaçúcares nos folíolos de populações F2 e F2RC1 (= F2 do primeiro retrocruzamento para Lycopersicon esculentum) do cruzamento interespecífico de L. esculentum TOM 584 (baixo teor de acilaçúcares) e L. pennellii LA 716 (alto teor). Foi realizado um teste de repelência ao ácaro T. evansi, tomando por base a distância percorrida pelos ácaros nos folíolos. Os genótipos selecionados para alto teor de acilaçúcares, em média, foram responsáveis por reduções significativas nas distâncias percorridas pelos ácaros sobre a superfície do folíolo. A repelência ao ácaro T. evansi exercida pelas plantas selecionadas para alto teor de acilaçúcares deu-se de forma semelhante àquela conferida pelo genitor LA-716. As magnitudes das correlações encontradas confirmam a associação entre altos teores do aleloquímico e a repelência ao ácaro.The Lycopersicon pennellii accession LA716 has been used as a source of arthropod pest resistance in tomato breeding programs due the high contents of acylsugars presents in its leaflets. We investigated the relationship between high foliar acylsugar contents and repellence to spider mite Tetranychus evansi in plants with contrasting acylsugar levels, selected from F2 and BC1F2 (= F2 of the first backcross towards L. esculentum) generations derived from the interespecific cross L. esculentum 'TOM-584' ;´; L. pennellii LA716. Mite resistance was assessed by a repellence test. Plants selected for high levels of acylsugars in leaflets had mite repellence levels similar to that of LA716. The high correlation confirmed the association between high acylsugar levels and mite repellence