In vitro determination of extracellular proteins from xylella fastidiosa

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The phytopathogen Xylella fastidiosa causes economic losses in important agricultural crops. Xylem vessel occlusion caused by biofilm formation is the major mechanism underlying the pathogenicity of distinct strains of X. fastidiosa. Here, we provide a detailed in vitro characterization of the extracellular proteins of X. fastidiosa. Based on the results, we performed a comparison with a strain J1a12, which cannot induce citrus variegated chlorosis symptoms when inoculated into citrus plants. We then extend this approach to analyze the extracellular proteins of X. fastidiosa in media supplemented with calcium. We verified increases in extracellular proteins concomitant with the days of growth and, consequently, biofilm development (330 days). Outer membrane vesicles carrying toxins were identified beginning at 10 days of growth in the 9a5c strain. In addition, a decrease in extracellular proteins in media supplemented with calcium was observed in both strains. Using mass spectrometry, 71 different proteins were identified during 30 days of X. fastidiosa biofilm development, including proteases, quorum-sensing proteins, biofilm formation proteins, hypothetical proteins, phage-related proteins, chaperones, toxins, antitoxins, and extracellular vesicle membrane components.The phytopathogen Xylella fastidiosa causes economic losses in important agricultural crops. Xylem vessel occlusion caused by biofilm formation is the major mechanism underlying the pathogenicity of distinct strains of X. fastidiosa. Here, we provide a de7Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2001/07533-7, 2012/51580-4]Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES, Computational Biology Program)CAPESFAPESP [2011/50268-4]CAPES (Computational Biology Program)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Spray-dried structured lipid carriers for the loading of Rosmarinus officinalis: new nutraceutical and food preservative

Rosemary, an aromatic herb with significant antioxidative activity, is frequently used as food preservative and a source of nutraceuticals. Its antioxidant effect is mainly related to the presence of phenolic compounds, molecules considerably unstable and prone to irreversible physicochemical changes when exposed to external agents. We here proposed the loading of rosemary into structured lipid systems to improve its physicochemical properties. Four formulations were prepared using the same amount of rosemary lyophilized extract. The lipid phase was composed of stearic acid and oleic acid, and the aqueous phase, a varying combination of drying carriers (whey protein concentrate or gum Arabic) and surfactant (Poloxamer 188). The formulations were sonicated, spray-dried, and the obtained powders were characterized regarding the density (0.18 g/mL to 0.26 g/mL), particle size distribution (7 µm and 52 µm), and water solubility (29% to 48%). The antioxidant activity was determined by applying ABTS+ radical-scavenging assay and the results expressed per gram of lyophilized extract (150.6 mol Trolox/g to 376.4 mol Trolox/g), with a significantly lower/higher result seen for formulations containing gum Arabic and a higher concentration of Poloxamer. The prepared systems may have potential applications as preservative in foodstuff and as nutraceutical.This research has been sponsored by CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) through the financial support (88887.368385/2019-00) granted to I.B., by the São Paulo ResearchFoundation (FAPESP) through the financial support (Grant N. 2018/26069-0 and 2011/10333-1) granted to W.P.O.and by the Portuguese Science and Technology Foundation (FCT) through the projects M-ERA-NET/0004/2015 (PAIRED) and UIDB/04469/2020 (strategic fund) granted to E.B.S.info:eu-repo/semantics/publishedVersio

Factors affecting the retention efficiency and physicochemical properties of spray dried lipid nanoparticles loaded with lippia sidoides essential oil

Essential oils (EOs) are widely used in various industrial sectors but can present several instability problems when exposed to environmental factors. Encapsulation technologies are effective solutions to improve EOs properties and stability. Currently, the encapsulation in lipid nanoparticles has received significant attention, due to the several recognized advantages over conventional systems. The study aimed to investigate the influence of the lipid matrix composition and spray-drying process on the physicochemical properties of the lipid-based nanoparticles loaded with Lippia sidoides EO and their retention efficiency for the oil. The obtained spray-dried products were characterized by determination of flow properties (Carr Index: from 25.0% to 47.93%, and Hausner ratio: from 1.25 to 1.38), moisture (from 3.78% to 5.20%), water activity (<0.5), and powder morphology. Zeta potential, mean particle size and polydispersity index, of the redispersed dried product, fell between 25.9 mV and 30.9 mV, 525.3 nm and 1143 nm, and 0.425 and 0.652, respectively; showing slight differences with the results obtained prior to spray-drying (from 16.4 mV to 31.6 mV; 147 nm to 1531 nm; and 0.459 to 0.729). Thymol retention in the dried products was significantly lower than the values determined for the liquid formulations and was affected by the drying of nanoparticles.I.B. acknowledges CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) for the financial support (88887.368385/2019-00). E.B.S. acknowledges the Portuguese Science and Technology Foundation (FCT) for the funded projects M-ERA-NET/0004/2015 (PAIRED) and UIDB/04469/2020 (strategic fund). WPO acknowledges the São Paulo Research Foundation (FAPESP) for the financial support (Grant N. 2018/26069-0 and 2011/10333-1).info:eu-repo/semantics/publishedVersio

Surface Physicochemical Properties At The Micro And Nano Length Scales: Role On Bacterial Adhesion And Xylella Fastidiosa Biofilm Development.

The phytopathogen Xylella fastidiosa grows as a biofilm causing vascular occlusion and consequently nutrient and water stress in different plant hosts by adhesion on xylem vessel surfaces composed of cellulose, hemicellulose, pectin and proteins. Understanding the factors which influence bacterial adhesion and biofilm development is a key issue in identifying mechanisms for preventing biofilm formation in infected plants. In this study, we show that X. fastidiosa biofilm development and architecture correlate well with physicochemical surface properties after interaction with the culture medium. Different biotic and abiotic substrates such as silicon (Si) and derivatized cellulose films were studied. Both biofilms and substrates were characterized at the micro- and nanoscale, which corresponds to the actual bacterial cell and membrane/ protein length scales, respectively. Our experimental results clearly indicate that the presence of surfaces with different chemical composition affect X. fastidiosa behavior from the point of view of gene expression and adhesion functionality. Bacterial adhesion is facilitated on more hydrophilic surfaces with higher surface potentials; XadA1 adhesin reveals different strengths of interaction on these surfaces. Nonetheless, despite different architectural biofilm geometries and rates of development, the colonization process occurs on all investigated surfaces. Our results univocally support the hypothesis that different adhesion mechanisms are active along the biofilm life cycle representing an adaptation mechanism for variations on the specific xylem vessel composition, which the bacterium encounters within the infected plant.8e7524

UM ESTUDO DE CASO: PALESTRA COMO FERRAMENTA DE CONSTRUÇÃO DO ENSINO E APRENDIZADO SOBRE “DESASTRES AMBIENTAIS” PARA REDUÇÃO DE RISCOS DE DESASTRES NA DISCIPLINA DE CIÊNCIAS

 O processo de desenvolvimento urbano sem planejamento, de parte considerável das cidades brasileiras, apresentam como consequência dois tipos de cidades. Uma cidade “formal” e uma cidade “informal”, oculta das estatísticas oficiais. A cidade informal se caracteriza por moradias irregulares, sem equipamentos urbanos básicos e situam-se em áreas que geralmente deveriam ser Áreas de Preservação Permanente (APP). Justamente estas áreas inapropriadas para ocupação, apresentam-se os maiores riscos de desastres ambientais, recorrentes no Vale do Itajaí. Neste sentido, a abordagem do conteúdo “Desastres Ambientais”, no ensino fundamental é de extrema importância como forma de educação para minimizar a suscetibilidade aos desastres futuros. O objetivo deste trabalho foi analisar o conhecimento construído pelos estudantes a partir de uma palestra ministrada pelo responsável da coordenação da Defesa Civil e aplicada aos estudantes de sexto ano do ensino fundamental. O conteúdo da palestra foi o tema Prevenção aos riscos Desastres Ambientais, com ênfase em deslizamento de solo. Essa prática docente ocorreu em uma escola da rede estadual do município de Brusque, em Santa Catarina. Após a palestra, os estudantes foram avaliados através de atividade de desenho, para representar duas situações: “casa em área de risco” e “casa fora da área de risco”. O resultado da análise indicou que praticamente todos os alunos apresentaram no desenho elementos relacionado à situação de risco. Os elementos identificados, em situação de risco foram moradias em encostas, aterros inadequados e a presença de chuva. Os elementos identificados em situação fora de risco foram residências em área planas, presença de vegetação e murros de proteção. Tendo visto a relevância do conteúdo “desastres ambientais” inserido no currículo do ensino fundamental, espera-se que práticas docentes tornem-se comuns nas escolas, visando a redução de riscos aos desastres

Vapd In Xylella Fastidiosa Is A Thermostable Protein With Ribonuclease Activity.

Xylella fastidiosa strain 9a5c is a gram-negative phytopathogen that is the causal agent of citrus variegated chlorosis (CVC), a disease that is responsible for economic losses in Brazilian agriculture. The most well-known mechanism of pathogenicity for this bacterial pathogen is xylem vessel occlusion, which results from bacterial movement and the formation of biofilms. The molecular mechanisms underlying the virulence caused by biofilm formation are unknown. Here, we provide evidence showing that virulence-associated protein D in X. fastidiosa (Xf-VapD) is a thermostable protein with ribonuclease activity. Moreover, protein expression analyses in two X. fastidiosa strains, including virulent (Xf9a5c) and nonpathogenic (XfJ1a12) strains, showed that Xf-VapD was expressed during all phases of development in both strains and that increased expression was observed in Xf9a5c during biofilm growth. This study is an important step toward characterizing and improving our understanding of the biological significance of Xf-VapD and its potential functions in the CVC pathosystem.10e014576

Spouted bed dried Rosmarinus officinalis extract: a novel approach for physicochemical properties and antioxidant activity

In this study, a conical-cylindrical spouted bed dryer with Teflon® beads as spouting material was used for producing powdered rosemary (Rosmarinus officinalis L.) extract. The influence of the inlet drying gas temperature (Tgi) and the percentage ratio between the feed rate of concentrated liquid extract by the maximum evaporation capacity of the spouted bed (Ws/Wmax) on selected physicochemical properties of the finished products were investigated. Antioxidant properties of the concentrated liquid extract and dried extracts were also evaluated by the 2.2-diphenyl-1-picrylhydrazyl radical scavenging (DPPH) and lipid peroxidation induced by Fe2+/citrate (LPO) methods; and compared with the values obtained for a lyophilized extract (used as a control). Colloidal silicon dioxide (Tixosil® 333) and maltodextrin (DE 14) at a 2:1 ratio was added to the concentrated extract before drying (4.4% w/w) to improve the drying performance. The drying variables Ws/Wmax and Tgi have statistically significant influence on total polyphenols and total flavonoid contents of the dried powders. The concentrated extract (on dry basisbeing absolute solid content) showed superior antioxidant activity (AA) compared to both the spouted bed dried and the lyophilized extracts; exhibiting IC50 values of 0.96 ± 0.02, 2.16 ± 0.04 and 3.79 ± 0.05 µg mL1 (DPPH method) and 0.22 ± 0.01, 1.31± 0.01 and 2.54 ± 0.02 µg mL1 (LPO method), respectively. These results of AA are comparable to values obtained for quercetin, a flavonoid compound often used as a reference standard due to its potent antioxidant activity; with IC50 of 1.17 µg mL1 (DPPH) and 0.22 µg mL1 (LPO). However, the dried rosemary extracts are about 13.5 times more concentrated than the initial concentrated extract (dry weight), with a concentration of total flavonoids and polyphenols compounds ranging from 4.3 to 12.3 and from 1.2 to 4.7 times higher than the concentrated extract values (wet basis). The AA per dry product mass was thus significantly higher than the values measured for concentrated extractive solution, irrespective of some losses of AA apparently due to the drying process.Foundation of Research Support of the São Paulo State (FAPESP) for the funded projects 2011/10333-1, 2012/03427-2 and 2018/26069-0 and for the National Council for Scientific and Technological Development (CNPq) for the financial support. E.B.S. acknowledges the Portuguese Science and Technology Foundation (FCT) for the funded projects M-ERA-NET/0004/2015 (PAIRED) and UIDB/04469/2020 (strategic fund)info:eu-repo/semantics/publishedVersio

DNA-interacting properties of two analogous square-planar cis-chlorido complexes: copper versus palladium

Two square-planar coordination compounds, namely [Cu(CPYA)Cl2] (1) and [Pd(CPYA)Cl2] (2), were prepared from the ligand 4-chloro-N-(pyridin-2-ylmethyl)aniline (CPYA) and two chloride salts, and were fully characterized, including by X-ray diffraction. Spectroscopic, electrophoretic and AFM studies revealed that the two isostructural compounds were interacting differently with DNA. In both cases, the initial interaction involves electrostatic contacts of the CPYA ligand in the minor groove (as suggested by molecular docking), but subsequent strong binding occurs with the palladium(II) complex 2, whereas the binding with the copper complex 1 is weaker and concentration dependent. The strong binding of 2 eventually leads to the cleavage of the double strand and the redox activity of 1 allows to oxidatively cleave the biomolecule

LaMMos - Latching Mechanism based on Motorized-screw for Reconfigurable Robots and Exoskeleton Suits

Reconfigurable robots refer to a category of robots that their components (individual joints and links) can be assembled in multiple configurations and geometries. Most of existing latching mechanisms are based on physical tools such as hooks, cages or magnets, which limit the payload capacity. Therefore, robots re- quire a latching mechanism which can help to reconfigure itself without sacrificing the payload capability. This paper presents a latching mechanism based on the flexible screw attaching principle. In which, actuators are used to move the robot links and joints while connecting them with a motorized-screw and dis- connecting them by unfastening the screw. The brackets used in our mechanism configuration helps to hold maximum force up to 5000N. The LaMMos - Latching Mechanism based on Motorized- screw has been applied to the DeWaLoP - Developing Water Loss Prevention in-pipe robot. It helps the robot to shrink its body to crawl into the pipe with minimum diameter, by recon- figuring the leg positions. And it helps to recover the legs positions to original status once the robot is inside the pipe. Also, LaMMos add stiffness to the robot legs by dynamically integrate them to the structure. Additionally, we present an application of the LaMMos mechanism to exoskeleton suits, for easing the mo- tors from the joints when carrying heavy weights for long periods of time. This mechanism offers many interesting opportunities for robotics research in terms of functionality, pay- load and size.Comment: 14 pages, 15 figure