985 resultados para Bimolecular fluorescence complementation


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Coffea arabica L. is considered to be sensitive to low temperatures throughout its life cycle. In some Brazilian regions, seedling production occurs under shade conditions and during the winter, with average temperatures of around 10 °C. The formation and functioning of the photosynthetic apparatus are strongly controlled by temperature. This study aimed to assess the changes that occurred in pigment contents, lipid peroxidation and variables of chlorophyll a fluorescence during the greening process of coffee seedlings submitted to chilling. Results indicate that saturation of the photosynthetic activity of coffee seedlings occurred before saturation of the accumulation of chloroplastid pigments. Pigment accumulation during the greening process is far beyond the metabolic needs for the maintenance of photosynthetic activity, more specifically of photosystem II. Coffee seedlings attained a quantum yield equivalent to that of the control with approximately half the chlorophyll a and b contents and around 40% of the carotenoid. Low temperature decreases the metabolism of seedlings, consequently reducing free radical production and lipid peroxidation. The chilling temperature (10 °C) used inhibited the accumulation of chloroplast pigments, in turn altering the capacity of the photosynthetic tissue of etiolated coffee seedlings to capture and transfer photon energy to the photosystem II reaction centre. These alterations were better demonstrated by O-J-I-P chlorophyll a fluorescence transients, rather than F v/F m and F v/F 0 ratios. © 2009 Elsevier B.V. All rights reserved.

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Optical characteristics of tellurite glasses containing silver nanoparticles (NPs) and the influence on the emission spectrum of Er 3+ ions were studied. The transitions 4f ↔ 4f from erbium ions, mainly the 4I13/2 → 4I15/2 transition that involve upconversion energy process, have a strongly dependence with the chemical structure of the rare earth ion. In the present work, silver nanparticles (NPs) embedded in the host vitreous material, show a significant enhance (or quenching) on the erbium fluorescence due the long-range electromagnetic interaction between the plasmon surface energy of the Ag NPs (Localized Surface Plasmon Resonance -LSPR) and the Er3+ ions.

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The aim of this study was to investigate the effects of some acidic drinks on dentin erosion, using methods of surface profile (SP) analysis and energy-dispersive X-ray fluorescence spectrometry (EDXRF). One hundred standardized dentin slabs obtained from bovine incisor roots were used. Dentin slabs measuring 5x5 mm were ground flat, polished and half of each specimen surface was protected with nail polish. For 60 min, the dentin surfaces were immersed in 50 mL of 5 different drinks (Gatorade®, Del Valle Mais orange juice®, Coca-Cola®, Red Bull® and white wine), 20 blocks in each drink. The pH of each beverage was measured. After the erosive challenge, the nail polish was removed and SP was analyzed. The mineral concentration of dentin surfaces was determined by means of EDXRF. Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). SP analysis showed that Red-Bull had the highest erosive potential (p<0.05). EDXRF results exhibited a decrease in phosphate in the groups immersed in Red-Bull, orange juice and white wine (p<0.05), and no significant difference in calcium content between the reference surface and eroded surface. In conclusion, this study demonstrated that all studied beverages promoted erosion on root dentin and Red Bull had the highest erosive potential. There was no correlation between pH of beverages and their erosive potential and only the P content changed after erosive challenge.

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The soluble lipase from Pseudomonas fluorescens (PFL) forms bimolecular aggregates in which the hydrophobic active centers of the enzyme monomers are in close contact. This bimolecular aggregate could be immobilized by multipoint covalent linkages on glyoxyl supports at pH 8.5. The monomer of PFL obtained by incubation of the soluble enzyme in the presence of detergent (0.5% TRITON X-100) could not be immobilized under these conditions. The bimolecular aggregate has two amino terminal residues in the same plane. A further incubation of the immobilized derivative under more alkaline conditions (e.g., pH 10.5) allows a further multipoint attachment of lysine (Lys) residues located in the same plane as the amino terminal residues. Monomeric PFL was immobilized at pH 10.5 in the presence of 0.5% TRITON X-100. The properties of both PFL derivatives were compared. In general, the bimolecular derivatives were more active, more selective and more stable both in water and in organic solvents than the monomolecular ones. The bimolecular derivative showed twice the activity and a much higher selectivity (100 versus 20) for the hydrolysis of R,S-2-hydroxy-4-phenylbutyric acid ethyl ester (HPBEt) in aqueous media at pH 5.0 compared to the monomeric derivative. In experiments measuring thermal inactivation at 75 °C, the bimolecular derivative was 5-fold more stable than the monomeric derivative (and 50-fold more stable than a one-point covalently immobilized PFL derivative), and it had a half-life greater than 4 h. In organic solvents (cyclohexane and tert-amyl alcohol), the bimolecular derivative was much more stable and more active than the monomeric derivative in catalyzing the transesterification of olive oil with benzyl alcohol. © 2012 Elsevier Ltd. All rights reserved.

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Two tomato scions (cvs. 'Raf' and 'Gorety') were grafted on three different rootstocks: S. torvum, 'Beaufort' (Lycopersicum esculentum × Lycopersicum hirsutum) and intermediate grafting of eggplant 'Cristal' between tomato and S. torvum (double graft). Plants were grown in Mediterranean greenhouse conditions. The response to grafting was measured through growth parameters, Fv/Fm and leaf macronutrients analysis, and it was compared with non-grafted plants. The scions grafted on S. torvum in simple and double graft showed lower fresh and dry weight of leaves, number of commercial fruits, plant height, Fv/Fm and decreased their capacity to absorb several nutrients resulting in a lower mineral concentration in scions leaves, as a result of a thickened graft union. On the other hand, both scions showed a good response when grafted on the rootstock 'Beaufort', with which growth parameters, yield and photosynthetic capacity were similar to non-grafted plants. © 2013 Copyright Taylor and Francis Group, LLC.

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Tuberculosis remains as one of the main cause of mortality worldwide due to a single infectious agent, Mycobacterium tuberculosis. The aroK-encoded M. tuberculosis Shikimate Kinase (MtSK), shown to be essential for survival of bacilli, catalyzes the phosphoryl transfer from ATP to the carbon-3 hydroxyl group of shikimate (SKH), yielding shikimate-3-phosphate and ADP. Here we present purification to homogeneity, and oligomeric state determination of recombinant MtSK. Biochemical and biophysical data suggest that the chemical reaction catalyzed by monomeric MtSK follows a rapid-equilibrium random order of substrate binding, and ordered product release. Isothermal titration calorimetry (ITC) for binding of ligands to MtSK provided thermodynamic signatures of non-covalent interactions to each process. A comparison of steady-state kinetics parameters and equilibrium dissociation constant value determined by ITC showed that ATP binding does not increase the affinity of MtSK for SKH. We suggest that MtSK would more appropriately be described as an aroL-encoded type II shikimate kinase. Our manuscript also gives thermodynamic description of SKH binding to MtSK and data for the number of protons exchanged during this bimolecular interaction. The negative value for the change in constant pressure heat capacity (ΔCp) and molecular homology model building suggest a pronounced contribution of desolvation of non-polar groups upon binary complex formation. Thermodynamic parameters were deconvoluted into hydrophobic and vibrational contributions upon MtSK:SKH binary complex formation. Data for the number of protons exchanged during this bimolecular interaction are interpreted in light of a structural model to try to propose the likely amino acid side chains that are the proton donors to bulk solvent following MtSK:SKH complex formation. © 2013 Rosado et al.

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The X-ray Fluorescence (XRF) analysis is a technique for the qualitative and quantitative determination of chemical constituents in a sample. This method is based on detection of the characteristic radiation intensities emitted by the elements of the sample, when properly excited. A variant of this technique is the Total Reflection X-ray Fluorescence (TXRF) that utilizes electromagnetic radiation as excitation source. In total reflection of X-ray, the angle of refraction of the incident beam tends to zero and the refracted beam is tangent to the sample support interface. Thus, there is a minimum angle of incidence at which no refracted beam exists and all incident radiation undergoes total reflection. In this study, we evaluated the influence of the energy variation of the beam of incident x-rays, using the MCNPX code (Monte Carlo NParticle) based on Monte Carlo method. © 2013 AIP Publishing LLC.

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In recent years, several studies have shown that concentrations of trace elements are altered in neoplastic breast tissues. However, the microenvironment and metabolic changes caused by tumors are complex and still not completely understood. Under this aspect, the combination of different techniques to investigate the role of trace elements in promoting and/or maintaining a tumor is interesting once the combination of information obtained by analytical techniques and immunohistochemical assays, associated with clinicopathological data, may allow a better metabolic understanding of trace elements in breast cancer. In this work, the role of the trace elements Ca, Fe, Cu and Zn in neoplastic breast tissues was investigated by X-ray fluorescence (XRF) techniques and immunohistochemical assays. We determined concentrations of Ca, Fe, Cu and Zn in normal and neoplastic breast tissues using energy dispersive XRF, and these values were used to set the positive or negative expression of elements in normal and neoplastic tissues. These expressions were correlated with the spatial distributions of trace elements (evaluated by micro-XRF) and with immunoexpression of matrix metalloproteinases (MMPs), tissue inhibitors of MMPs and vascular endothelial growth factor. The results revealed that the expression of the trace elements Fe, Cu and vascular endothelial growth factor are related, indicating that higher levels of these elements can be associated with the angiogenic process in breast cancer. Also, associations between Ca, Zn and MMPs expression have been observed, possibly because of the fact that both metals are present in these proteins. © 2013 John Wiley & Sons, Ltd.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Pigmentation patterns, ultraviolet reflection and fluorescent emission are often involved in mate recognition and mate quality functions in many animal taxa. We investigated the role of wing ultra-violet reflection, fluorescence emission, and pigmentation on age and sexual signals in the damselfly Mnesarete pudica. In this species, wings are sexually dimorphic in colour and exhibit age dependency: males and females show a smoky black colouration when young, turning red in mature males while it turns brown in females. First, we investigated wing UV patterns through reflectance and emission spectra. Second, behavioural experiments were undertaken to show male and female responses to manipulated wing pigmentation and experimentally reduced UV (UV-). Reflectance spectra of the wings of juvenile and mature males and females were used to show the differences between controls and individuals with manipulated colouration used in the behavioural experiment. UV-reduced, females with wings painted red, and control males and females were tethered and presented to conspecific males and females, and their behavioral responses were recorded. The male red wing pigmentation and females with red wings elicited an aggressive response in territorial males and a sexual response in females. Both males and females showed neutral responses towards individuals with reduced UV. Wing signals of juvenile individuals also provoked neutral responses. These results suggest that UV, together with pigmentation, plays a role during mate recognition in males and females. Other than butterflies and spiders, it seems that fluorescence signals and UV reflectance can also be part of communication in odonates. © 2013 Springer Science+Business Media New York.

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In cases of identification of bones, skeletal segments or isolated bones, searching for biotypologic diagnostic data to estimate an individual's age enables comparing these data with those of missing individuals. Enamel, dentin and pulp undergo remarkable changes during an individual's life. The enamel becomes more mineralized, smoother and thinner, and deteriorates because of physiological and pathological factors. Dental pulp decreases in volume due to the deposition of secondary dentin; thus, the dentin becomes thicker with time. In natural teeth, the fluorescence phenomenon occurs in dentin and enamel and changes in those tissues may alter the expression of the natural tooth color. The aim of this study was to assess the correlation between age and teeth fluorescence for individuals from different age groups. The sample consisted of 66 randomly selected Brazilians of both genders aged 7-63 years old. They were divided into 6 groups: Group 1 - aged 7-12 years, Group 2 - aged 13-20 years, Group 3 - aged 21-30 years, Group 4 - aged 31-40 years, Group 5 - aged 41-50 years and Group 6 - aged between 51 and 63 years. Upper right or left central incisors were used for the study. Restored and aesthetic rehabilitated teeth were excluded from the sample. The measurement of tooth fluorescence was carried out via computer analysis of digital images using the software ScanWhite DMC/Darwin Systems - Brazil. It was observed that dental fluorescence decreases when comparing the age groups 21-30, 31-40, 41-50 and 51-63 years. The results also showed that there is a statistically significant difference between the groups 41-50 years and 21-30 years (p=. 0.005) and also among the group 51-63 years and all other groups (p< 0.005). It can be concluded that dental fluorescence is correlated with age and has a similar and stable behavior from 7 to 20 years of age. It reaches its maximum expected value at the age of 26.5 years and thereafter decreases. © 2013 Elsevier B.V.

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The objective of this study was to optimize an internal control to improve SYBR-Green-based qPCR to amplify/detect the BoHV-5 US9 gene in bovine embryos produced invitro and experimentally exposed to the virus. We designed an SYBR-Green-based binding assay that is quick to perform, reliable, easily optimized and compares well with the published assay. Herein we demonstrated its general applicability to detect BoHV-5 US9 gene in bovine embryos produced invitro experimentally exposed to BoHV-5. In order to validate the assay, three different reference genes were tested; and the histone 2a gene was shown to be the most adequate for normalizing the qPCR reaction, by considering melting and standard curves ( p<0.05). On the other hand, no differences were found in the development of bovine embryos invitro whether they were exposed to BoHV-5 reference and field strains comparing to unexposed embryos. The developed qPCR assay may have important field applications as it provides an accurate BoHV-5 US9 gene detection using a proven reference gene and is considerably less expensive than the TaqMan qPCR currently employed in sanitary programs. © 2013 Elsevier Ltd.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)