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Biophysical methods are widely employed in academia and the pharmaceutical industry to detect and quantify weak molecular interactions. Such methods find broad application in fragment-based drug discovery (FBDD). In an FBDD campaign, a suitable affinity determination method is key to advancing a project beyond the initial screening phase. Protein-observed (PO) nuclear magnetic resonance (NMR) finds widespread use due to its ability to sensitively detect very weak interactions at residue-level resolution. When there are issues precluding the use of PO-NMR, ligand-observed (LO) NMR reporter assays can be a useful alternative. Such assays can measure affinities in a similar range to PO-NMR while offering some distinct advantages, especially with regard to protein consumption and compound throughput. In this paper, we take a closer look at setting up such assays for routine use, with the aim of getting high-quality, accurate data and good throughput. We assess some of the key characteristics of these assays in the mathematical framework established for fluorescence polarization assays with which the readers may be more familiar. We also provide guidance on setting up such assays and compare their performance with other affinity determination methods that are commonly used in drug discovery.

CsPbX3 perovskite nanoplates (PNPLs) were formed in a synthesis driven by SnX4 (X=Cl, Br, I) salts. The role played by these hard Lewis acids in directing PNPL formation is addressed. Sn4+ disturbs the acid-base equilibrium of the system, increasing the protonation rate of oleylamine and inducing anisotropic growth of nanocrystals. Sn4+ cations influence the reaction dynamics owing to complexation with oleylamine molecules. By monitoring the photoluminescence excitation and photoluminescence (PL) spectra of the PNPLs grown at different temperatures, the influence of the thickness on their optical properties is mapped. Time-resolved and spectrally resolved PL for colloidal dispersions with different optical densities reveals that the dependence of the overall PL lifetime on the emission wavelength do not originate from energy transfer between PNPLs but from the contribution of PNPLs with distinct thickness, indicating that thicker PNPLs exhibit longer PL lifetimes.

Although regular combined aerobic-resistance exercises can ameliorate the inflammatory status and redox balance in elderly population, it is unclear whether protein or specific amino acid supplementation could improve such benefits. Therefore, we aimed to evaluate the inflammatory status and redox indexes through of the saliva of 34 elderly subject nonpractitioners (NP group, 73.3±6.6 years) and 49 elderly subject practitioners of a combined-exercise training in moderate intensity (CET group, 71.9±5.8 years) before (pre) and after (post) 30 days of supplementation with L-glutamine (Gln) or placebo (PL). Our results showed that, both in pre- and postsupplementation, the salivary levels of nitric oxide (NO⋅) and TNF-α were lower, whereas the levels of uric acid and IL-10 (as well as IL-10/TNF-α ratio) were higher in the CET groups than in the NP groups. In postsupplementation, both groups supplemented with Gln (NP-Gln and CET-Gln) showed higher salivary uric acid levels compared to baseline. In addition, lower NO⋅ levels were found in the CET-Gln group postsupplementation than presupplementation values. Whereas the CET-Gln group showed lower GSH levels postsupplementation, NP-Gln subjects showed lower GSSG levels at the same time point, both compared to baseline. Interestingly, salivary peroxidase activity was lower only in NP groups (NP-PL and NP-Gln) postsupplementation than baseline values. A positive significant correlation between salivary peroxidase activity and GSH levels, and also between salivary peroxidase activity and uric acid levels were observed in the CET-Gln group both pre- and postsupplementation. No differences were found in albumin, total antioxidant activity (TEAC), and reducing power analysis between groups, pre- or postsupplementation. In conclusion, the elderly subjects from the CET group showed a better inflammatory response and redox balance and, for the first time, it was shown that daily supplementation with Gln for 30 days can improve these benefits with putative association with a healthy aging.

Closely spaced promoters in tandem formation are abundant in bacteria. We investigated the evolutionary conservation, biological functions, and the RNA and single-cell protein expression of genes regulated by tandem promoters in E. coli. We also studied the sequence (distance between transcription start sites ‘dTSS’, pause sequences, and distances from oriC) and potential influence of the input transcription factors of these promoters. From this, we propose an analytical model of gene expression based on measured expression dynamics, where RNAP-promoter occupancy times and dTSS are the key regulators of transcription interference due to TSS occlusion by RNAP at one of the promoters (when dTSS ≤ 35 bp) and RNAP occupancy of the downstream promoter (when dTSS > 35 bp). Occlusion and downstream promoter occupancy are modeled as linear functions of occupancy time, while the influence of dTSS is implemented by a continuous step function, fit to in vivo data on mean single-cell protein numbers of 30 natural genes controlled by tandem promoters. The best-fitting step is at 35 bp, matching the length of DNA occupied by RNAP in the open complex formation. This model accurately predicts the squared coefficient of variation and skewness of the natural single-cell protein numbers as a function of dTSS. Additional predictions suggest that promoters in tandem formation can cover a wide range of transcription dynamics within realistic intervals of parameter values. By accurately capturing the dynamics of these promoters, this model can be helpful to predict the dynamics of new promoters and contribute to the expansion of the repertoire of expression dynamics available to synthetic genetic constructs.Author SummaryTandem promoters are common in nature, but investigations on their dynamics have so far largely relied on synthetic constructs. Thus, their regulation and potentially unique dynamics remain unexplored. We first performed a comprehensive exploration of the conservation of genes regulated by these promoters in E. coli and the properties of their input transcription factors. We then measured protein and RNA levels expressed by 30 Escherichia coli tandem promoters, to establish an analytical model of the expression dynamics of genes controlled by such promoters. We show that start site occlusion and downstream RNAP occupancy can be realistically captured by a model with RNAP binding affinity, the time length of open complex formation, and the nucleotide distance between transcription start sites. This study contributes to a better understanding of the unique dynamics tandem promoters can bring to the dynamics of gene networks and will assist in their use in synthetic genetic circuits.

We studied the evolution of the incubation effect during fs-laser micromachining of gallium nitride films with the number of laser pulses applied per sample surface area. The damage threshold fluence was measured using the zero-damage method and reduced from (0.61 ± 0.01) J/cm2 to (0.05 ± 0.03) J/cm2 when increasing the number of applied laser pulses from single-shot excitation to 105 pulses per spot, respectively. By applying the exponential defect accumulation model to the experimental incubation data, we determined a small value for the incubation parameter of (0.011 ± 0.001), indicating that gallium nitride has slow incubation dynamics. This information is important to achieve the desired control and high efficiency of fs-laser processing of gallium-nitride-based samples.

Luminescent semiconductor nanocrystals or quantum dots (QDs) emerge as important fluorescent probes for in vitro and in vivo Trypanosoma cruzi cells studies. However, to ensure applicability to living organisms, several tests still need to be done. Since several toxic events are caused by QDs, such as loss of mitochondrial membrane potential, ROS generation, DNA damage and cell death by autophagy. We performed a review of the literature on mechanisms of cellular uptake, internalization and citotoxicity of nanoparticles including our results about the evaluation of biological toxicity in T. cruzi. We evaluated the possible effects on parasite growth curves in a time - scale of control and incubated cells with different concentrations of CdTe – QDs (0.2; 2.0; 20 and 200µM) to determine the development cells changes. In addition, intracellular ROS were measured by Electron Paramagnetic Resonance Spectroscopy (EPR) technique. According our results, we can infer that the toxic effects of QDs in T. cruzi are dose-dependent and that high levels of ROS are involved in cellular toxicity promoted by higher concentrations of QDs. In summary, parasites labeled with low concentrations of nanoparticles are suitable and can be used as bioimaging tools for living parasites. However, more studies on QDs cytotoxicity need to be carried out. Los nanocristales semiconductores luminiscentes o puntos cuánticos (QD) emergen como importantes sondas fluorescentes para estudios in vitro e in vivo con células de Trypanosoma cruzi. Sin embargo, para garantizar la aplicabilidad a los organismos vivos, aún deben realizarse varias pruebas. Dado que varios eventos tóxicos son causados por QD, como la pérdida del potencial de la membrana mitocondrial, la generación de ROS, el daño del ADN y la muerte celular por autofagia. Realizamos una revisión de la literatura sobre los mecanismos de captación celular, internalización y citotoxicidad de las nanopartículas, incluidos nuestros resultados sobre la evaluación de la toxicidad biológica en T. cruzi. Evaluamos los posibles efectos sobre las curvas de crecimiento del parásito en una escala de tiempo de control y las células incubadas con diferentes concentraciones de CdTe - QDs (0.2; 2.0; 20; 200µM) para determinar los cambios en las células de desarrollo. Además, las ROS intracelulares se midieron mediante la técnica de espectroscopia de resonancia paramagnética electrónica (EPR). Según nuestros resultados, podemos inferir que los efectos tóxicos de las QD en T. cruzi son dosis-dependientes y que los niveles altos de ROS están implicados en la toxicidad celular promovida por concentraciones más altas de QD. En resumen, los parásitos etiquetados con bajas concentraciones de nanopartículas son adecuados y pueden usarse como herramientas de bioimagen para parásitos vivos. Sin embargo, es necesario realizar más estudios sobre citotoxicidad QD. Nanocristais semicondutores luminescentes ou pontos quânticos (QDs) surgem como importantes sondas fluorescentes para estudos in vitro e in vivo de células de Trypanosoma cruzi. No entanto, para garantir a aplicabilidade aos organismos vivos, vários testes ainda precisam ser feitos. Uma vez que diversos eventos tóxicos são causados por QDs, como perda de potencial de membrana mitocondrial, geração de ROS, dano ao DNA e morte celular por autofagia. Realizamos uma revisão da literatura sobre os mecanismos de captação celular, internalização e citotoxicidade de nanopartículas incluindo nossos resultados sobre a avaliação da toxicidade biológica em T. cruzi. Avaliamos os possíveis efeitos nas curvas de crescimento do parasita em uma escala de tempo de controle e células incubadas com diferentes concentrações de CdTe - QDs (0,2; 2,0; 20 e 200µM) para determinar as alterações nas células de desenvolvimento. Além disso, as ROS intracelulares foram medidas pela técnica de Espectroscopia de Ressonância Paramagnética Eletrônica (EPR). De acordo com nossos resultados, podemos inferir que os efeitos tóxicos dos QDs no T. cruzi são dose-dependentes e que altos níveis de ROS estão envolvidos na toxicidade celular promovida por maiores concentrações de QDs. Em resumo, parasitas marcados com baixas concentrações de nanopartículas são adequados e podem ser usados como ferramentas de bioimagem para parasitas vivos. No entanto, mais estudos sobre citotoxicidade de QDs precisam ser realizados.

Induced pluripotent stem cells (iPSC) have been the focus of several studies due to their wide range of application, including in cellular therapy. The use of iPSC in regenerative medicine is limited by their tumorigenic potential. Extracellular vesicles (EV) derived from stem cells have been shown to support renal recovery after injury. However, no investigation has explored the potential of iPSC-EV in the treatment of kidney diseases. To evaluate this potential, we submitted renal tubule cells to hypoxia-reoxygenation injury, and we analyzed cell death rate and changes in functional mitochondria mass. An in vivo model of ischemia-reperfusion injury was used to evaluate morphological and functional alterations. Gene array profile was applied to investigate the mechanism involved in iPSC-EV effects. In addition, EV derived from adipose mesenchymal cells (ASC-EV) were also used to compare the potential of iPSC-EV in support of tissue recovery. The results showed that iPSC-EV were capable of reducing cell death and inflammatory response with similar efficacy than ASC-EV. Moreover, iPSC-EV protected functional mitochondria and regulated several genes associated with oxidative stress. Taken together, these results show that iPSC can be an alternative source of EV in the treatment of different aspects of kidney disease.

Boron-based glasses are relevant materials for optical applications on account its ability to hold large amount of oxide modifiers and rare-earth ions. The barium borates ones are particularly interesting due to its designed second and third-order optical nonlinearities. Despite the interest on barium borate glasses for linear and nonlinear optics, the fabrication of active elements for optical microcircuits are still scare. In this paper, we demonstrate the fabrication of three dimensional waveguides in barium borate glass with broad emission in the visible spectrum. Such performance was achieved through the co-doping with Dy 3 + and Eu 3 + ions, which together provide broad emissions at blue, yellow and red, along with the glass processing with femtosecond laser pulses. Almost circular waveguides, with diameter of approximately 5 μ m and 7.8 mm long, were inscribed into the bulk of the glass by direct laser writing, whose support single mode guiding at 632.8 nm with propagation loss of (3 ± 1) dB/cm. When coupling UV light, such waveguides display the typical emission of Dy 3 + and Eu 3 + ions with its additional guiding throughout the waveguide length. The broad band emission and its guiding in the visible spectrum revel that the waveguides fabricated by femtosecond laser micromachining in co-doped barium borate glass are promising active elements for the development of optical micro-devices.

This paper presents a study on the incubation effect during fs-laser micromachining of gallium nitride films with three different wavelengths (1030, 515 and 343 nm) by varying the number of laser pulses applied per sample surface area and measuring the damage threshold fluence using the zero damage method. As we implemented the exponential defect accumulation model to the experimental incubation data, we determined the value of the incubation parameter of (0.02 ± 0.01) for 1030 nm, (0.07 ± 0.01) for 515 nm and (0.4 ± 0.1) for 343 nm. This shows that for excitation in the green and infrared spectral region, GaN requires approximately 100 overlapping pulses in order for the incubation to take place while, for ultraviolet excitation, the incubation was achieved faster, with the overlap of about 10 pulses. Furthermore, we compared our data for the single pulse damage threshold with a theoretical model that takes into account multiphoton and avalanche ionization. Our results indicate that at 343 nm and 515 nm, micromachining is dominated by multiphoton ionization, while at 1030 nm other effects, such as tunneling ionization, must also be contributing. Finally, we believe this paper brings relevant information on the fs-laser micromachining of GaN.

Borate glasses present the ability to host various modifier ions and, therefore, are adaptable for a wide range of applications. Specifically, calcium-lithium tetraborate (CaLiBO) glasses doped with rare earths have been investigated for applications involving energy transfer processes. In this work, waveguides were inscribed by femtosecond laser microfabrication in CaLiBO glasses containing Tb3+ and Yb3+ ions, displaying emission in the green region. Single-mode guiding at 632.8 nm, with propagation losses of approximately 2.0 dB/cm, was obtained in 7.0-mm long waveguides with a diameter on the order of 2 μm, produced in the glass bulk, at approximately 100 μm below the sample surface. By coupling 488 nm light, the fabricated waveguide showed the capability of guiding the typical emission of Tb3+ ions. Such glasses containing rare earths could be suitable for the development of green light-emitting microdevices.

Although essential for inflammatory responses, leukocyte recruitment to blood vessel walls in response to inflammatory stimuli, such as TNF-α, can contribute to vascular occlusion in inflammatory diseases, including atherosclerosis. We aimed to further characterize the mechanisms by which TNF stimulates adhesive and morphologic alterations in neutrophils. Microfluidic and intravital assays confirmed the potent effect that TNF has on human and murine neutrophil adhesion and recruitment in vitro and in vivo, respectively. Inhibition of actin polymerization by cytochalasin D significantly diminished TNF-induced human neutrophil adhesion in vitro and abolished TNF-induced membrane alterations and cell spreading. In contrast, TNF-induced increases in β2-integrin (Mac-1 and LFA-1) expression was not significantly altered by actin polymerization inhibition. Consistent with a role for cytoskeletal rearrangements in TNF-induced adhesion, TNF augmented the activity of the Rho GTPase, RhoA, in human neutrophils. However, inhibition of the major RhoA effector protein, Rho kinase (ROCK), by Y-27632 failed to inhibit TNF-induced neutrophil adhesion. In contrast, the formin FH2 domain inhibitor, SMIFH2, abolished TNF-induced human neutrophil adhesion and diminished leukocyte recruitment in vivo. SMIFH2 also inhibited TNF-induced cytoskeletal reorganization in human neutrophils and abolished the alterations in β2-integrin expression elicited by TNF stimulation. As such, Rho GTPase/mDia formin-mediated cytoskeletal reorganization appears to participate in the orchestration of TNF-induced neutrophil-adhesive interactions, possibly mediated by formin-mediated actin nucleation and subsequent modulation of β2-integrin activity on the neutrophil surface. This pathway may represent a pharmacologic target for reducing leukocyte recruitment in inflammatory diseases.

The complex pathophysiology of sickle cell anemia (SCA) involves intravascular hemolytic processes and recurrent vaso-occlusion, driven by chronic vascular inflammation, which result in the disease's severe clinical complications, including recurrent painful vaso-occlusive episodes. Hydroxyurea, the only drug frequently used for SCA therapy, is a cytostatic agent, although it appears to exert nitric oxide/soluble guanylyl cyclase (sGC) modulating activity. As new drugs that can complement or replace the use of hydroxyurea are sought to further reduce vaso-occlusive episode frequency in SCA, we investigated the effects of the sGC agonists BAY 60-2770 (sGC activator) and BAY 41-2272 (sGC stimulator) in the presence or absence of hydroxyurea on SCA vaso-occlusive mechanisms and cell recruitment both ex vivo and in vivo. These agents significantly reduced stimulated human SCA neutrophil adhesive properties ex vivo in association with the inhibition of surface β2-integrin activation. A single administration of BAY 60-2770 or BAY 41-2272 decreased tumor necrosis factor cytokine-induced leukocyte recruitment in a mouse model of SCA vaso-occlusion. Importantly, the in vivo actions of both agonists were significantly potentiated by the coadministration of hydroxyurea. Erythroid cell fetal hemoglobin (HbF) elevation is also a major goal for SCA therapy. BAY 41-2272 but not BAY 60-2770 at the concentrations employed significantly induced γ-globin gene transcription in association with HbF production in cultured erythroleukemic cells. In conclusion, sGC agonist drugs could represent a promising approach as therapy for SCA, for use either as stand-alone treatments or in combination with hydroxyurea. SIGNIFICANCE STATEMENT: This preclinical study demonstrates that stimulators and activators of sGC are potent inhibitors of the adhesion and recruitment of leukocytes from humans and in mice with sickle cell anemia (SCA) and may represent a promising approach for diminishing vaso-occlusive episode frequency in SCA. Hydroxyurea, a drug already frequently used for treating SCA, was found to potentiate the beneficial effects of sGC agonists in in vivo studies, implying that these classes of compounds could be used alone or in combination therapy.

Background: Although it has been previously demonstrated that acute inflammation can promote the tumor growth of a sub-tumorigenic dose of melanoma cells through of 5-lipoxygenase inflammatory pathway and its product leukotriene B4, and also that the peritumoral treatment with eicosapentaenoic acid and its product, leukotriene B5, reduces the tumor development, the effect of the treatment by gavage with omega-3 and omega-6 in the tumor microenvironment favorable to melanoma growth associated with acute inflammation has never been studied. Methods: C57BL/6 mice were coinjected with 1 &times, 106 apoptotic cells plus 1 &times, 103 viable melanoma cells into the subcutaneous tissue and treated by gavage with omega-3-rich fish oil or omega-6-rich soybean oil or a mixture of these oils (1:1 ratio) during five consecutive days. Results: The treatment by gavage with a mixture of fish and soybean oils (1:1 ratio) both reduced the melanoma growth and the levels of leukotriene B4 (LTB4), prostaglandin E2 (PGE2), PGE2/prostaglandin E3 (PGE3) ratio, and CXC ligand 1 (CXCL1) and increased the levels of interleukin 10 (IL-10) to IL-10/CXCL1 ratio in the melanoma microenvironment. Conclusion: The oral administration of a 1:1 mixture of fish oil and soybean oil was able to alter the release of inflammatory mediators that are essential for a microenvironment favorable to the melanoma growth in mice, whereas fish oil or soybean oil alone was ineffective.

This study aimed to optimize Cymbopogon citratus essential oil loaded into PLGA-nanoparticles by investigating the effect of processing variables (sonication time, ultrasound power, and essential oil/polymer ratio) on encapsulation efficiency and particle mean hydrodynamic diameter using Box-Behnken design. Nanoparticles were prepared by an emulsification/solvent diffusion method and physicochemically characterized by FTIR, DSC and TGA/DTA. Cytotoxicity was evaluated in human HaCat keratinocytes by WST-1 and LDH assays. The optimized formulation had a hydrodynamic mean diameter of 277 nm, a polydispersity index of 0.18, a Zeta potential of −16 mV and an encapsulation efficiency of 73%. Nanoparticle characterization showed that only citral was incorporated in nanocarriers, with some amount adsorbed on their surface, and highlighted the potential in increasing the oil thermal stability. The drug release profile demonstrated a biphasic pattern with a substantial sustained release depending on diffusion from the polymeric matrix. Toxicity effects on cell viability of pure essential oil at low concentrations were significantly eliminated when encapsulated. Results revealed the ability of PLGA-nanoparticles to improve essential oil physicochemical characteristics, by controlling release and reducing toxicity, suggesting their potential use in pharmaceutical preparations.

Xylanases catalyze the hydrolysis of β-1,4-linked xylosyl moieties from xylan chains, one of the most abundant hemicellulosic polysaccharides found in plant cell walls. These enzymes can exist either as single catalytic domains or as modular proteins composed of one or more carbohydrate-binding modules (CBMs) appended to the catalytic core. However, the molecular mechanisms governing the synergistic effects between catalytic domains and their CBMs are not fully understood. Thus, the goal of this study was to evaluate the functional effects of the fusion of a CBM belonging to family 6, which exhibits high affinity to xylan, with the GH11 xylanase from Bacillus subtilis, which does not have a CBM in its wild-type form. The wild-type enzyme (BsXyl11) and the chimeric protein (BsXyl11-CBM6) were heterologously produced in Escherichia coli and purified to homogeneity for biochemical characterization. The molecular fusion did not alter the pH and temperature dependence, but kinetic data revealed an increase of 65% in the catalytic efficiency of the chimeric enzyme. Furthermore, the BsXyl11-CBM6 chimera was used to supplement the commercial cocktail Accellerase® 1500 and improved the reducing sugar release by 17% from pretreated sugarcane bagasse. These results indicate that CBM6 can be used as a molecular tool to enhance the catalytic performance of endo-xylanases (GH11) and provide a new strategy for the development of optimized biocatalysts for biotechnological applications.

Capillarity is the study of interfaces between two immiscible liquids or between a liquid and a vapor. The theory of capillarity was created in the early 1800s, and it is applicable to mesoscopic and macroscopic (>1 μm) systems. In general, macroscopic theories are expected to fail at the

Made available in DSpace on 2020-12-12T02:38:43Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-15 Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Agencia Nacional de Promoción Científica y Tecnológica Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) In this study was synthesized cerium oxide (CeO2) doped with praseodymium (Pr) using a microwave-assisted hydrothermal method. The positions of the vacancies with respect to Ce3+/Ce4+ and the vacancy-like defects surrounded by the electronic density of Pr atoms were determined through photoluminescence (PL) measurements, positron annihilation lifetime spectroscopy, and density functional theory (DFT). Distinct PL responses were observed for Pr-doped CeO2 indicating that oxygen vacancies contribute to the formation of deep energy levels in the forbidden region, thus facilitating charge transfer. The interplay between the experimental measurements and computational simulations at the microscopic level based on DFT revealed the charge rearrangement in oxygen-deficient CeO2:Pr systems. These results indicate that oxygen vacancies and electrons at the 4f states contribute to electrical conduction, thereby demonstrating that Pr-doped CeO2 acts as an n-type semiconductor. Faculty of Engineering of Guaratinguetá São Paulo State University – UNESP Permanent Address:Modeling and Molecular Simulations Group São Paulo State University UNESP Department of Physics Federal University of Sao Carlos Laboratório Interdisciplinar de Materiais Avançados Universidade Federal de Itajubá Unifei – Campus CDMF LIEC Chemistry Department of the Federal University of São Carlos - (UFSCar), P.O. Box 676 Institute of Materials Science and Technology (INTEMA) University of Mar del Plata and National Research Council (CONICET), Juan B. Justo 4302 Instituto de Física de Materiales Tandil UE CIFICEN (CONICET- Consejo Nacional de Investigaciones Científicas y Técnicas-CONICET), Pinto 399 Faculty of Engineering of Guaratinguetá São Paulo State University – UNESP Permanent Address:Modeling and Molecular Simulations Group São Paulo State University UNESP CAPES: 001 FAPESP: 2008/57872-1 Agencia Nacional de Promoción Científica y Tecnológica: 2015–1832 FAPESP: 2018/20729-9 FAPESP: 2019/08928-9 CNPq: 573636/2008-7

Made available in DSpace on 2020-12-12T01:08:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-07-15 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Nd(OH)3-ZnO composites were synthesized with three content of Neodymium (1, 3 and 5 wt%) and characterized in terms of their structural, morphological, vibrational, optical, surface, photocatalytic and antimicrobial properties. The Nd(OH)3-ZnO composites are constituted by ZnO, which showed hexagonal structure, three-dimensional morphology analogous to an arrangement of flowers with hexagonal rods, and the Nd(OH)3 in the hexagonal phase with irregularly spherical shape. The presence of the Nd(OH)3 in the composites promoted some structural degree of disorder in the ZnO network as well as interstitial defects and oxygen vacancy, which improved photocatalytic and antimicrobial activity. From the Box-Behnken design and response surface methodology, it was determined that the 1Nd(OH)3-ZnO composite showed the highest photocatalytic activity, which was attributed to its higher crystallinity, smaller crystallite size and particle width, band gap energy close to ZnO and lower recombination rate compared to ZnO. Nd(OH)3-ZnO composites showed higher antimicrobial activity against E. coli and S. aureus than ZnO pure, evidencing the synergistic effect between Nd(OH)3 and ZnO in the composites. Therefore, this study highlights the synthesis of Nd(OH)3-ZnO composites combining zinc oxide and neodymium hydroxide with excellent photocatalytic and antimicrobial activity, which can be used in different practical applications such as luminescent material, photocatalyst and antimicrobial agent. Program of Post-Graduation in Chemical Engineering State University of West Parana, P.O. Box 520 CDMF LIEC Federal University of São Carlos, P.O. Box 676 Modeling and Molecular Simulations Group São Paulo State University UNESP Modeling and Molecular Simulations Group São Paulo State University UNESP

Sugarcane is a very important crop in the history of Brazil, since the period of colonization. Among the factors that affect its yield in commercial areas, weeds could be highlighted. Appropriate management strategies are essential to mitigate this interference and increase crop productivity. In this sense, the aim was to evaluate the effect of different control methods on weed management, in the pre-emergence of two varieties, of cane-plant. The experiment was conducted at Primavera Farm, Itambé (PE), in randomized blocks, in a 2x5 factorial scheme, with 4 replications. The first factor represented two varieties of sugarcane (RB867515 and RB92579) and the second five weed control methods (weed control, absolute control, s-metalochlor + tebuthiuron, sulfentrazone and oxyfluorfen). The number of tillers per linear meter, degree of phytotoxicity and weed control were evaluated. At the end of the research, the RB92579 variety can be used as an alternative for the weed control because it presents rapid sprouting and initial development and more aggressive tillering than RB867515, under Itambé (PE) soil conditions. It is recommended the use of sulfentrazone at the dose of 1.8 L.ha-1.p.c, for better residual effect and "sufficient" control of weeds in the stage of establishment of the culture.

This study aimed to prepare hydrogel containing Cymbopogon citratus (DC.) Stapf, Poaceae, volatile oil encapsulated in poly (d,l-lactide-co-glycolide) nanoparticles and to evaluate its in vitro anti-herpetic activity. Polymeric nanoparticles were prepared by solvent emulsification-diffusion method and incorporated in carbomer hydrogels. In vitro release profiles for the nanogel, loaded nanoparticles and hydrogel containing free oil were evaluated by dialysis. Inhibitory activities against Herpes simplex for the formulations were investigated in Vero cells. Hydrogel was developed using nanoparticles with mean diameter of 217.1 nm and negative Zeta potential (−20.5 mV). Volatile oil release profile showed a biphasic pattern with an initial faster release and subsequent sustained phase in all formulations. Nanogel strongly inhibited virus in a non-cytotoxic concentration, 42.16 times lower than free oil, 8.76 and 2.23 times than loaded nanoparticles and hydrogel containing free oil, respectively. These results highlight the potential of nanogel to protect oil against volatilization, control release and improve its anti-herpetic activity. Keywords: Nanotechnology, Polymeric nanoparticles, Volatile oil, Lemongrass, Hydrogel, Herpes simplex virus

In this paper, we further investigate the properties of off-stoichiometry thiol-ene polymers (OSTE) aiming its application in microchip electrophoresis for bioanalytical applications. The proportion of 1.3:1 (allyl:thiol) and 1:2.5 (allyl:thiol) presented the best results in terms of sealing. Raman imaging mapping of the polymers surfaces revealed an outstanding homogeneity. Water contact angle were measured as 68° ± 6° and 71° ± 5° for 1.3:1 allyl and 1:2.5 thiol, respectively. Substrates prepared with OSTE demonstrated to be less prone to sorption of nonpolar compounds. The electroosmotic flow measured for this OSTE composition was 3.8 ± 0.3·10–4 cm2 s–1 V–1, 1.5 times higher than the one found for polydimethylsiloxane microchips under the same conditions. As a proof-of-concept for the applicability of OSTE microchips in bioanalysis the immobilization of α-amylase on the polymer surface and the implementation of a Saccharomyces cerevisiae cell counter using contactless conductivity detection are demonstrated.

Resumo A cevada (Hordeum vulgare) é o principal cereal utilizado na produção de cerveja, mas é importada, na maioria dos países tropicais e subtropicais. Isto implica no aumento do custo de produção da bebida, além de não satisfazer a demanda das bebidas sem glúten no mercado. O sorgo (Sorghum bicolor L. Moench) é um dos cereais que também podem ser usados na produção de cerveja e, ao contrário da cevada, é livre de glúten, recomendado para doentes celíacos. Assim, recentemente tem aumentado o interesse no uso do malte de sorgo para substituir o malte de cevada na produção de bebidas. No entanto, as principais dificuldades encontradas na produção de bebidas fermentadas a partir de malte de sorgo têm sido o baixo nível de enzimas hidrolíticas e a alta temperatura de gelatinização. O objetivo deste trabalho de revisão foi avaliar os principais fatores que afetam os processos de maltagem (tempo e temperatura de imersão, de germinação e de secagem) e a mosturação com malte de sorgo (pH e temperatura das enzimas, e métodos usados) na produção de cerveja sem glúten. Abstract Barley (Hordeum vulgare) is the main cereal used in beer production, but is imported in most of the tropical and subtropical countries. This increases the production costs, without satisfying the market demand for gluten-free beverages. Sorghum ( Sorghum bicolor L. Moench) is one of the cereals that can also be used in brewing, and, unlike barley, is gluten-free and hence recommended for celiac patients. Thus interest in the use of sorghum malt to replace barley malt in beverage production has increased recently. However, the main difficulties encountered in the production of fermented beverages using sorghum malt have been the low level of hydrolytic enzymes and high gelatinization temperature. Hence the objective of this review was to evaluate the main factors affecting malting processes (drying and germination, steeping time and temperature) and mashing with sorghum malt (pH and temperature of the enzymes and methods used) in gluten-free beer production.

As hypoxia-induced inflammatory angiogenesis may contribute to the manifestations of sickle cell disease, we compared the angiogenic molecular profiles of plasma from sickle cell disease individuals and correlated these with in vitro endothelial cell-mediated angiogenesis-stimulating activity and in vivo neovascularization. Bioplex demonstrated that plasma from patients with steady-state sickle cell anemia contained elevated concentrations of pro-angiogenic factors (angiopoietin-1, basic fibroblast growth factor, vascular endothelial growth factor, vascular endothelial growth factor-D and placental growth factor) and displayed potent pro-angiogenic activity, significantly increasing endothelial cell proliferation, migration and capillary-like structure formation. In vivo neovascularization of Matrigel plugs was significantly greater in sickle cell disease mice than in non-sickle cell disease mice, consistent with an up-regulation of angiogenesis in the disease. In plasma from patients with hemoglobin SC disease without proliferative retinopathy, anti-angiogenic endostatin and thrombospondin-2 were significantly elevated. In contrast, plasma from hemoglobin SC individuals with proliferative retinopathy had a pro-angiogenic profile and more significant effects on endothelial cell proliferation and capillary formation than plasma from patients without retinopathy. Hydroxyurea therapy was associated with significant reductions in plasma angiogenic factors and inhibition of endothelial cell-mediated angiogenic mechanisms and neovascularization. Thus, individuals with sickle cell anemia or hemoglobin SC disease with retinopathy present a highly angiogenic circulating milieu, capable of stimulating key endothelial cell-mediated angiogenic mechanisms. Combination anti-angiogenic therapy to prevent the progression of unregulated neovascularization and associated manifestations in sickle cell disease, such as pulmonary hypertension, may be indicated; furthermore, the benefits and drawbacks of the potent anti-angiogenic effects of hydroxyurea should be clarified.

This work investigates the modification, resulting from fs-laser irradiation (150 fs, 775 nm and 1 kHz), on the structure and surface morphology of hydrogenated amorphous silicon (a-Si:H) thin films. The sample morphology was studied by performing a statistical analyzes of atomic force microscopy images, using a specially developed software that identifies and characterizes the domains (spikes) produced by the laser irradiation. For a fluence of 3.1 MJ/m2, we observed formation of spikes with smaller average height distribution, centered at around 15 nm, while for fluencies higher than 3.7 MJ/m2 aggregation of the produced spikes dominates the sample morphology. On the other hand, Raman spectroscopy revealed that a higher crystalline fraction (73%) is obtained for higher fluences (> 3.1 MJ/m2), which is accompanied by a decrease in the size of the produced crystals. Therefore, such results indicate that there is a trade-off between the spike distribution, crystallization fraction and size of the nanocrystals attained by laser irradiation, which has to be taken into account when using such approach for the development of devices.

We resolve the phonon structure from from lowest exciton transition of CdSe/ZnS quantum dots ensemble by separating the homogeneous and inhomogeneous linewidths through optical multidimensional coherent spectroscopy at low temperatures.

This study evaluated the use of three solid brewery wastes: brewer's spent grain, hot trub and residual brewer's yeast, as alternative media for the cultivation of lactic acid bacteria to evaluate their potential for proteolytic enzyme production. Initially, a mixture experimental design was used to evaluate the effect of each residue, as well as different mixtures (with the protein content set at 4%) in the enzyme production. At predetermined intervals, the solid and liquid fractions were separated and the extracellular proteolytic activity was determined. After selecting the best experimental conditions, a second experiment, factorial experimental design, was developed in order to evaluate the protein content in the media (1 to 7%) and the addition of fermentable sugar (glucose, 1 to 7%). Among the wastes, residual yeast showed the highest potential for the production of extracellular enzymes, generating a proteolytic extract with 2.6 U/mL in 3 h. However, due to the low content of the fermentable sugars in the medium, the addition of glucose also had a positive effect, increasing the proteolytic activity to 4.9 U/mL. The best experimental conditions of each experimental design were reproduced for comparison, and the enzyme content was separated by ethanol precipitation. The best medium produced a precipitated protein with proteolytic activity of 145.5 U/g.

The current demand for fabricating optical and photonic devices displaying high performance, using low-cost and time-saving methods, prompts femtosecond (fs)-laser processing as a promising methodology. High and low repetition femtosecond lasers enable surface and/or bulk modification of distinct materials, which can be used for applications ranging from optical waveguides to superhydrophobic surfaces. Herein, some fundamental aspects of fs-laser processing of materials, as well as the basics of their most common experimental apparatuses, are introduced. A survey of results on polymer fs-laser processing, resulting in 3D waveguides, electroluminescent structures and active hybrid-microstructures for luminescence or biological microenvironments is presented. Similarly, results of fs-laser processing on glasses, gold and silicon to produce waveguides containing metallic nanoparticles, analytical chemical sensors and surface with modified features, respectively, are also described. The complexity of fs-laser micromachining involves precise control of material properties, pushing ultrafast laser processing as an advanced technique for micro/nano devices.

Inhibition of leukocyte adhesion to the vascular endothelium represents a novel and important approach for decreasing sickle cell disease (SCD) vaso-occlusion. Using a humanized SCD–mouse-model of tumor necrosis factor-α–induced acute vaso-occlusion, we herein present data demonstrating that short-term administration of either hydroxyurea or the phosphodiesterase 9 (PDE9) inhibitor, BAY73-6691, significantly altered leukocyte recruitment to the microvasculature. Notably, the administration of both agents led to marked improvements in leukocyte rolling and adhesion and decreased heterotypic red blood cell-leukocyte interactions, coupled with prolonged animal survival. Mechanistically, these rheologic benefits were associated with decreased endothelial adhesion molecule expression, as well as diminished leukocyte Mac-1-integrin activation and cyclic guanosine monophosphate (cGMP)–signaling, leading to reduced leukocyte recruitment. Our findings indicate that hydroxyurea has immediate beneficial effects on the microvasculature in acute sickle-cell crises that are independent of the drug's fetal hemoglobin-elevating properties and probably involve the formation of intravascular nitric oxide. In addition, inhibition of PDE9, an enzyme highly expressed in hematopoietic cells, amplified the cGMP-elevating effects of hydroxyurea and may represent a promising and more tissue-specific adjuvant therapy for this disease.

The increasing interest of consumers in health and alcohol abuse issues motivates breweries to expand the assortment of products with low alcohol content. The goal of producing beers with low alcohol content can be achieved by two main strategies; namely by gentle removal of alcohol from regular beer and by limited ethanol formation during the beer fermentation. Within these two basic strategies, there are a number of techniques that vary in performance, efficiency and usability. This paper presents an overview and comparison of these techniques and provides an evaluation of sensorial properties of low-alcohol and an alcohol-free beer produced as well as suggests possibilities for their additional improvement.

Quantum dots (QDs) are colloidal semiconductor nanocrystals of a few nanometers in diameter, being their size and shape controlled during the synthesis. They are synthesized from atoms of group II–VI or III–V of the periodic table, such as cadmium telluride (CdTe) or cadmium selenium (CdSe) forming nanoparticles with fluorescent characteristics superior to current fluorophores. The excellent optical characteristics of quantum dots make them applied widely in the field of life sciences. Cellular uptake of QDs, location and translocation as well as any biological consequence, such as cytotoxicity, stimulated a lot of scientific research in this area. Several studies pointed to the cytotoxic effect against micoorganisms. In this mini-review, we overviewed the synthesis and optical properties of QDs, and its advantages and bioapplications in the studies about microorganisms such as protozoa, bacteria, fungi and virus.

The principal aim of studies of enzyme-mediated reactions has been to provide comparative and quantitative information on enzyme-catalyzed reactions under distinct conditions. The classic Michaelis–Menten model (Biochem Zeit 49:333, 1913) for enzyme kinetic has been widely used to determine important parameters involved in enzyme catalysis, particularly the Michaelis–Menten constant (K M ) and the maximum velocity of reaction (V max ). Subsequently, a detailed treatment of the mechanisms of enzyme catalysis was undertaken by Briggs–Haldane (Biochem J 19:338, 1925). These authors proposed the steady-state treatment, since its applicability was constrained to this condition. The present work describes an extending solution of the Michaelis–Menten model without the need for such a steady-state restriction. We provide the first analysis of all of the individual reaction constants calculated analytically. Using this approach, it is possible to accurately predict the results under new experimental conditions and to characterize and optimize industrial processes in the fields of chemical and food engineering, pharmaceuticals and biotechnology.

Higher efficiency processes are strongly dependent on the ability to process materials/ components that can sustain more aggressive service conditions. b-NiAl intermetallic alloys are known for exhibiting superior high temperature properties, but are difficult to process. This study evaluated Ni–Al coatings processed in situ by plasma transferred arc (PTA) welding. Coatings processed with mixtures of Ni and Al powders, Al content within the range 15–50 wt-%, were analysed. In situ development of Ni aluminides was confirmed by X-ray diffraction analysis. Increasing aluminium (Al) content in the deposited mixture increased the penetration depth. The microstructure of coatings depended on the amount of Al in the deposited powder mixture associated with the exothermal synthesis of the b-NiAl intermetallic compound. The present study presents an important step towards the protection of components for equipment of higher efficiency processes, as results validated the potential of PTA hardfacing as a processing technique for surface tailoring.

Leukocyte adhesion to vessel walls may initiate vaso-occlusion in sickle cell anemia (SCA); however, the extent to which inflammation participates in this mechanism is not understood. This in vitro study investigated whether inflammatory molecules, commonly augmented in SCA, can affect neutrophil adhesive properties and whether cyclic guanosine monophosphate (cGMP)-elevating agents can inhibit such adhesion.Effects of Interleukin 8 (IL-8), tumor necrosis factor-α (TNF-α), granulocyte macrophage-colony stimulating factor (GM-CSF) cytokines, BAY 73-6691 [phosphodiesterase (PDE)-9A-inhibitor], and BAY 41-2271 (guanylate-cylase stimulator) on the adhesive properties of neutrophils from healthy control (CON) and steady-state SCA individuals were determined using static-adhesion assays.SCA neutrophils demonstrated increased adhesive properties, compared to CON neutrophils; IL-8, TNF-α and GM-CSF increased CON neutrophil adhesion and further increased SCA neutrophil adhesion to fibronectin (FN). The PDE9A inhibitor, BAY-73-6691, significantly reduced basal CON neutrophil and SCA neutrophil adhesion; this was accompanied by decreased SCA neutrophil surface expressions of the L-selectin and CD11b adhesion molecules. BAY-73-6691 also significantly reduced cytokine-stimulated CON neutrophil and SCA neutrophil adhesion to FN; however, this was not accompanied by alterations in adhesion-molecule presentation.The chronic inflammatory nature of SCA may contribute to leukocyte adhesive functions in SCA. Furthermore, elevation of leukocyte cGMP may be an interesting approach for inhibition of leukocyte adhesion to the vessel wall, even in the presence of inflammatory stimuli.

Oxidative stress plays a significant role in sickle cell disease (SCD), contributing to haemolysis, vaso-occlusive processes and endothelial dysfunction. To study the effects that the serum of SCD individuals has on the oxidative state of blood cells, sera were pooled from control individuals, steady-state SCD patients and SCD patients on hydroxyurea therapy (SCDHU), and their effects on markers of oxidative stress and damage in neutrophils isolated from healthy individuals observed. Incubation of control neutrophils, but not platelets nor red blood cells, with SCD serum (10% v/v; 2 hours) significantly augmented their production of reactive oxygen species (ROS). Increased ROS production in SCD serum-incubated neutrophils was associated with increased superoxide anion generation, apoptosis and increased nicotinamide adenine dinucleotide phosphate oxidase subunit expression. Although serum from SCDHU individuals also induced ROS generation in neutrophils, its oxidative capacity appeared to be lower. Results suggest that factors in the serum of SCD individuals contribute to ROS generation and oxidative damage in leukocytes.

The main aim of this work was to produce fruit wines from pulp of gabiroba, cacao, umbu, cupuassu and jaboticaba and characterize them using gas chromatography–mass spectrometry for determination of minor compounds and gas chromatography-flame ionization detection for major compounds. Ninety-nine compounds (C6 compounds, alcohols, monoterpenic alcohols, monoterpenic oxides, ethyl esters, acetates, volatile phenols, acids, carbonyl compounds, sulfur compounds and sugars) were identified in fruit wines. The typical composition for each fruit wine was evidenced by principal component analysis and Tukey test. The yeast UFLA CA 1162 was efficient in the fermentation of the fruit pulp used in this work. The identification and quantification of the compounds allowed a good characterization of the fruit wines. With our results, we conclude that the use of tropical fruits in the production of fruit wines is a viable alternative that allows the use of harvest surpluses and other underused fruits, resulting in the introduction of new products into the market., Conselho Nacional de Desenvolvimento Científico e Tecnológico do Brasil (CNPq) and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior)

Homogeneous mixtures of Ce0.8Gd0.2O1.9 (GDC) and La0.6Sr0.4Co0.2Fe0.8O3 (LSCF) nanopowders were successfully synthesized using induction plasma by axial injection of a solution. The resulting nanocomposite powders consisted of two kinds of nanopowders with different mass ratio of GDC/LSCF, such as 3/7 and 6/4. The morphological features, crystallinity, and the phases of the synthesized powders were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), local energy-dispersive x-ray spectroscopy (EDS) analysis, and x-ray diffraction (XRD). The nanopowders are almost globular in shape with a diameter smaller than 100 nm and their BET specific areas are around 20 m2 g−1. The GDC and LSCF phases are well distributed in the nanopowders. In addition, suspensions, made with the as-synthesized composite nanopowders and ethanol, were used to deposit cathode coatings using suspension plasma spray (SPS). Micro-nanostructures of the coatings are discussed. The coatings are homogeneous and porous (51% porosity) with cauliflower structures.

Avaliou-se o fracionamento dos carboidratos e das proteínas das gramíneas tropicais tifton 85 (Cynodon dactylon cv Tiffon 85), braquiária (Brachiaria brizantha cv Marandu) e tanzânia (Panicum maximum Jacq. cv Tanzânia), cortadas aos 28, 35 e 54 dias de idade. Determinaram-se os teores de matéria seca (MS), matéria orgânica (MO), proteína bruta (PB), extrato etéreo (EE), fibra em detergente neutro corrigida para cinzas e proteína (FDN CP), fibra em detergente ácido (FDA), lignina (LIG), nitrogênio insolúvel em detergente neutro (NIDN) e nitrogênio insolúvel em detergente ácido (NIDA). Para PB determinaram-se as frações A, B1+B2, B3 e C, e para os carboidratos as frações A+B1, B2 e C. Foi utilizado o delineamento inteiramente ao acaso, em arranjo em parcelas (gramíneas) subdivididas (idades de corte). Houve efeito significativo (P

The objective of this study was to evaluate the ethanol production from the sugars contained in the sugarcane bagasse hemicellulosic hydrolysate with the yeast Pichia stipitis DSM 3651. The fermentations were carried out in 250-mL Erlenmeyers with 100 mL of medium incubated at 200 rpm and 30 degrees C for 120 h. The medium was composed by raw (non-detoxified) hydrolysate or by hydrolysates detoxified by pH alteration followed by active charcoal adsorption or by adsorption into ion-exchange resins, all of them supplemented with yeast extract (3 g/L), malt extract (3 g/L), and peptone (5 g/L). The initial concentration of cells was 3 g/L. According to the results, the detoxification procedures removed inhibitory compounds from the hemicellulosic hydrolysate and, thus, improved the bioconversion of the sugars into ethanol. The fermentation using the non-detoxified hydrolysate led to 4.9 g/L ethanol in 120 h, with a yield of 0.20 g/g and a productivity of 0.04 g L(-1) h(-1). The detoxification by pH alteration and active charcoal adsorption led to 6.1 g/L ethanol in 48 h, with a yield of 0.30 g/g and a productivity of 0.13 g L(-1) h(-1). The detoxification by adsorption into ion-exchange resins, in turn, provided 7.5 g/L ethanol in 48 h, with a yield of 0.30 g/g and a productivity of 0.16 g L(-1) h(-1).

BACKGROUND: This work deals with the xylitol production by biotechnological routes emphasizing the purification process using crystallization. RESULTS: Xylitol volumetric productivity of 0.665 g L −1 h −1 and yield of 0.7024 g g −1 were obtained after 92 h fermentation. The fermented broth (61.3 g L −1 xylitol) was centrifuged, treated and concentrated obtain a syrup (745.3 g L −1 xylitol) which was crystallized twice, xylitol crystals with 98.5 – 99.2% purity being obtained. CONCLUSION: The hypothetical distribution obtained permits the determination of modeling parameters, which make possible the estimation of crystal dominant size from different initial experimental conditions. c � 2008 Society of Chemical Industry

Banana, an important component in the diet of the global population, is one of the most consumed fruits in the world. This fruit is also very favorable to industry processes (e.g., fermented beverages) due to its rich content on soluble solids and minerals, with low acidity. The main objective of this work was to evaluate the influence of factors such as banana weight and extraction time during a hot aqueous extraction process on the total soluble solids content of banana. The extract is to be used by the food and beverage industries. The experiments were performed with 105 mL of water, considering the moisture of the ripe banana (65%). Total sugar concentrations were obtained in a beer analyzer and the result expressed in degrees Plato (°P, which is the weight of the extract or the sugar equivalent in 100 g solution at 20 °C), aiming at facilitating the use of these results by the beverage industries. After previous studies of characterization of the fruit and of ripening performance, a 22 full-factorial star design was carried out, and a model was developed to describe the behavior of the dependent variable (total soluble solids) as a function of the factors (banana weight and extraction time), indicating as optimum conditions for extraction 38.5 g of banana at 39.7 min., Gabinete de Relações Internacionais da Ciência e do Ensino Superior (GRICES), Portugal., Fundação de Amparo à Pesquisa do Estado de São Paulo (Fapesp), Brasil., Coordenação para Aperfeiçoamento do Ensino Superior (CAPES), Brasil., Fundação para a Ciência e Tecnologia (FCT), Portugal.

Nývlt method was used to determine the kinetic parameters of commercial xylitol in ethanol:water (50:50 %w/w) solution by batch cooling crystallization. The kinetic exponents ( n , g and m ) and the system kinetic constant ( B N ) were determined. Model experiments were carried out in order to verify the combined effects of saturation temperatures (40, 50 and 60 °C) and cooling rates (0.10, 0.25 and 0.50 °C/min) on these parameters. The fitting between experimental and calculated crystal sizes has 11.30% mean deviation.

Modulation of intracellular cyclic guanosine monophosphate (cGMP) may characterize a therapeutic target for sickle cell disease (SCD); cGMP-dependent signalling may be important for erythroid foetal haemoglobin induction and exert anti-inflammatory functions in leucocytes. As the inhibition of phosphodiesterases (PDEs), which regulate intracellular cGMP, can result in tissue-specific elevation of cGMP, we studied the gene expressions of cGMP-specific PDEs (-1A, -5A and -9A) in the reticulocytes and neutrophils of healthy controls, steady-state SCD patients and SCD patients on hydroxycarbamide therapy (SCDHC). PDE9A gene expression was found in numerous cell types; however, high expression was found in neutrophils, reticulocytes, CD34(+)-derived erythroid cells and K562 erythroleukaemic cells, indicating a high haematopoietic cell expression. PDE9A gene expression was, however, significantly higher in the reticulocytes and neutrophils of SCD individuals, compared to control cells; Western blotting confirmed the production of PDE9A protein in SCD neutrophils and K562 cells. Inhibition of PDE9A enzyme with the specific inhibitor, BAY73-6691, significantly increased production of the gamma-globin gene (HBG) in K562 cells and reversed the increased adhesive properties of SCD neutrophils. Since elevation of haematopoietic intracellular cGMP may be beneficial in SCD, the relatively limited tissue distribution of PDE9A suggests that it could represent a novel drug target worthy of further study.

BACKGROUND: Xylitol bioproduction from lignocellulosic residues comprises hydrolysis of the hemicellulose, detoxification of the hydrolysate, bioconversion of the xylose, and recovery of xylitol from the fermented hydrolysate. There are relatively few reports on xylitol recovery from fermented media. In the present study, ion-exchange resins were used to clarify a fermented wheat straw hemicellulosic hydrolysate, which was then vacuum-concentrated and submitted to cooling in the presence of ethanol for xylitol crystallization. RESULTS: Sequential adsorption into two anion-exchange resins (A-860S and A-500PS) promoted considerable reductions in the content of soluble by-products (up to 97.5%) and in medium coloration (99.5%). Vacuum concentration led to a dark-colored viscous solution that inhibited xylitol crystallization. This inhibition could be overcome by mixing the concentrated medium with a commercial xylitol solution. Such a strategy led to xylitol crystals with up to 95.9% purity. The crystallization yield (43.5%) was close to that observed when using commercial xylitol solution (51.4%). CONCLUSION: The experimental data demonstrate the feasibility of using ion-exchange resins followed by cooling in the presence of ethanol as a strategy to promote the fast recovery and purification of xylitol from hemicellulose-derived fermentation media. Copyright © 2008 Society of Chemical Industry

Sickle cell disease (SCD) is a chronic inflammatory condition characterized by high leucocyte counts, altered cytokine levels and endothelial cell injury. As the removal of inflammatory cells by apoptosis is fundamental for the resolution of inflammation, we aimed to determine whether the leucocyte apoptotic process is altered in SCD. Neutrophils from SCD individuals showed an inhibition of spontaneous apoptosis when cultured in vitro, in the presence of autologous serum for 20 h. Intracellular cyclic adenosine monophosphate (cAMP) levels were approximately twofold increased in SCD neutrophils; possible cAMP-upregulating factors present in SCD serum include interleukin-8, granulocyte-macrophage colony-stimulating factor and prostaglandin. Accordingly, co-incubation of SCD neutrophils with KT5720, a cAMP-dependent protein kinase (PKA) inhibitor, abrogated increased SCD neutrophil survival. Caspase-3 activity was also significantly diminished in SCD neutrophils cultured for 16 h and this activity was restored when cells were co-incubated with KT5720. BIRC2 (encoding cellular inhibitor of apoptosis protein 1, cIAP(1)), MCL1 and BAX expression were unaltered in SCD neutrophils; however, BIRC3 (encoding the caspase inhibitor, cIAP(2)), was expressed at significantly higher levels. Thus, we report an inhibition of spontaneous SCD neutrophil apoptosis that appears to be mediated by upregulated cAMP-PKA signalling and decreased caspase activity. Increased neutrophil survival may have significant consequences in SCD; contributing to leucocytosis, tissue damage and exacerbation of the chronic inflammatory state.

The solubility of d-glucose in water as a function of temperature in a mixture of ethanol/water containing (50, 60, 70, and 80) % ethanol was evaluated on the basis of the refraction index at 60 °C. The results obtained have shown that the solubility in water presents a linear increase with an increase in temperature, whereas for the solutions containing ethanol/water mixtures, this increase is represented by a second-order polynomial. Because glucose shows considerably high solubility, these data can be used to study the glucose crystallization process in mixtures containing antisolvents.

This work deals with the development of a technology for xylitol production from sugarcane hemicellulosic hydrolysate by biotechnological routes emphasizing the purification process by crystallization. The hemicellulosic hydrolysate obtained by acid hydrolysis of sugarcane bagasse, was concentrated and treated with ion-exchange resins. The fermentation (88 g/L xylose, pH 5.0, 30 h−1 oxygen transfer volumetric coefficient, 30 °C and 300 rpm stirring rate) was carried out in a 15-L bioengineering fermentator. The fermented broth was centrifuged, treated, concentrated and subsequently crystallized twice. This process resulted in xylitol in crystals with 92–94% purity.

J. Inst. Brew. 113(4), 391–398, 2007 The present work evaluated the influence of all-malt wort orig-inal gravity on fermentative parameters and flavour-active com-pound formation during primary fermentation of high gravity brewing by a continuous process using a lager yeast immobilised on a natural carrier obtained from brewer’s spent grain (the main brewery by-product). The all-malt worts with original gravity (OG) ranging from 13.4 to 18.5°Plato were prepared by diluting a very-high-gravity wort (20°Plato) with sterile brewery water. The continuous assay was carried out in a bubble column bio-reactor with a total working volume of 5.2 litres, at 15°C, using a constant gas flow rate of 250 mL/min (200 mL/min of CO 2 and 50 mL/min of air) and a dilution rate of 0.04 h -1 (residence time of 25 h). The results indicated that as the wort OG was in-creased, the ethanol concentration of the outflowing beer in-creased. On the other hand, the continuous fermentation of the most concentrated worts (16.6 and 18.5°Plato) resulted in beers with unbalanced flavour profiles due to excessive ethyl acetate formation. The immobilised cell concentration appeared to be nearly independent from increasing wort OG.

One of the most important properties of quantum dots (QDs) is their size. Their size will determine optical properties and in a colloidal medium their range of interaction. The most common techniques used to measure QD size are transmission electron microscopy (TEM) and X-ray diffraction. However, these techniques demand the sample to be dried and under a vacuum. This way any hydrodynamic information is excluded and the preparation process may alter even the size of the QDs. Fluorescence correlation spectroscopy (FCS) is an optical technique with single molecule sensitivity capable of extracting the hydrodynamic radius (HR) of the QDs. The main drawback of FCS is the blinking phenomenon that alters the correlation function implicating in a QD apparent size smaller than it really is. In this work, we developed a method to exclude blinking of the FCS and measured the HR of colloidal QDs. We compared our results with TEM images, and the HR obtained by FCS is higher than the radius measured by TEM. We attribute this difference to the cap layer of the QD that cannot be seen in the TEM images.