Enzymatic Activity and Its Relationships with the Total Phenolic Content and Color Change in the High Hydrostatic Pressure-Assisted Curing of Vanilla Bean (Vanilla planifolia).

Diverse enzymatic reactions taking place after the killing of green vanilla beans are involved in the flavor and color development of the cured beans. The effects of high hydrostatic pressure (HHP) at 50-400 MPa/5 min and blanching as vanilla killing methods were evaluated on the total phenolic content (TPC), polyphenoloxidase (PPO), and peroxidase (POD) activity and the color change at different curing cycles of sweating-drying (C0-C20) of vanilla beans. The rate constants describing the above parameters during the curing cycles were also obtained. The TPC increased from C1 to C6 compared with the untreated green beans after which it started to decrease. The 400 MPa samples showed the highest rate of phenolic increase. Immediately after the killing (C0), the highest increase in PPO activity was observed at 50 MPa (46%), whereas for POD it was at 400 MPa (25%). Both enzymes showed the maximum activity at C1, after which the activity started to decrease. As expected, the L* color parameter decreased during the entire curing for all treatments. An inverse relationship between the rate of TPC decrease and enzymatic activity loss was found, but the relationship with L* was unclear. HHP appears to be an alternative vanilla killing method; nevertheless, more studies are needed to establish its clear advantages over blanching.

Interaction of triblock copolymers (Pluronic®) with DMPC vesicles: a photophysical and computational study.

Pluronic/lipid mix promises stealth liposomes with long circulation time and long-term stability for pharmaceutical applications. However, the influence of Pluronics on several aspects of lipid membranes has not been fully elucidated. Herein it was described the effect of Pluronics on the structured water, alkyl chain conformation, and kinetic stability of dimyristoylphosphatidylcholine (DMPC) liposomes using interfacial and deeper fluorescent probes along with computational molecular modeling data. Interfacial water changed as a function of Pluronics' hydrophobicity with polypropylene oxide (PPO) anchoring the copolymers in the lipid bilayer. Pluronics with more than 30-40 PO units had facilitated penetration at the bilayer while shorter PPO favored a more interfacial interaction. Low Pluronic concentrations provided long-term stability of vesicles by steric effects of polyethylene oxide (PEO), but high amounts destabilized the vesicles as a sum of water-bridge cleavage at the polar head group and the reduced alkyl-alkyl interactions among the lipids. The high kinetic stability of Pluronic/DMPC vesicles is a proof-of-concept of its advantages and applicability in nanotechnology over conventional liposome-based pharmaceutical products for future biomedical applications.

Fluorescent organic particle doped polymer-based gel dosimeter for neutron detection.

The purpose of this work is to develop a material capable of detecting neutrons produced by photodisintegration in a linear accelerator for its medical use. In this study, we have developed a gel-like material doped with fluorescent organic particles. PPO at 1 wt% is used as primary dopant and POPOP as secondary one at 0.03 wt%. A set of four samples is produced, with boric acid concentrations of 0, 400, 800 and 1200 ppm. The viscoelastic properties of the material are characterized with rheological measurements, finding a gel-like behavior, i.e., a material that can keep its original shape if no stresses are applied, but can also be deformed by applying a moderate shear rate. Furthermore, the material was irradiated with gamma, electron, and neutron emission sources from 137Cs, 22Na, 60Co, 210Po, 90Sr and 241AmBe, and its response was measured in two different experimental settings, in two different institutions, for comparative purposes. From these measurements, one can clearly establish that the new material detects neutrons, electrons, and gammas within the MeV regions and below. Thus, our findings show that the developed material and its properties make it a promising technology for its use in a neutron detector.

Phosphate Fertilization as a Modulator of Enzymatic Browning in Minimally Processed Cassava.

This study proposes to relate the increase in phosphorus (P) supply in the soil, via phosphate fertilization, to oxidative damage and protection, phenylpropanoid metabolism, and enzymatic browning in minimally processed cassava. The roots were grown with 0, 60, and 120 kg ha-1 P2O5. The roots were harvested, and the yield and P content in the root, stem, and leaves were quantified. The roots were minimally processed and stored for 12 days at 5 °C. The higher supply of P in the soil increased the P content of roots and stems but not the agro-industrial yields. Roots grown at 120 kg ha-1 P2O5 showed higher detection of hydrogen peroxide, which was accompanied by increased phenolic compounds, soluble quinones, and antioxidant capacity and increased activities of the enzymes superoxide dismutase, catalase, ascorbate peroxidase, polyphenol oxidase, and peroxidase. The present study thus demonstrates the role of phosphorus application, induction of the synthesis of phenolic compounds, and quality of fresh-cut cassava.

Morphophysiological characteristics of Brazilian bean genotypes related with sulfentrazone tolerance.

In the bean crop there are limited options of herbicides to control eudicotyledons weeds in pre-emergence and sulfentrazone presents potential to be used in a weed management program. The aim of this study was to determine the tolerance of a large number of Brazilian bean genotypes to sulfentrazone and their relation with morphophysiological characteristics, associating it to market classes, seed size and their center of origin. It was evaluated the effect of sulfentrazone application (400 g a.i. ha-1) in pre-emergence of 40 Brazilian bean genotypes, 36 of which were common-bean (Phaseolus vulgaris), two cowpeas (Vigna unguiculata), one adzuki bean (Vigna angularis) and one mung bean (Vigna radiata). There was high tolerance variation of the genotypes to sulfentrazone. Morphological parameters were strongly related with relative tolerance. Common bean genotypes that had medium to large seed size (cranberry and carioca) were more tolerant, while small-sized seed genotypes were more sensitive (black). Andean genotypes were more tolerant than Mesoamerican genotypes. Within the Mesoamerican group, medium-sized seed genotypes (carioca beans) were more tolerant than small-sized seed (black beans). Considering only the Vigna genera, cowpea was highly tolerant to sulfentrazone, while mung bean showed intermediary response and adzuki bean was completely sensible.

Weed control and soybean agronomic performance in response to application of sulfentrazone + diuron in pre-emergence / Controle de plantas daninhas e desempenho agronômico de soja em resposta à aplicação de sulfentrazone + diuron em pré-emergência

The use of pre-emergent herbicides is an important tool in weedmanagement. Sulfentrazone + diuron mixture was recently launched ontothe market. The objectiveof this work was to evaluate the weed control and the agronomic performance of soybean in response to application of sulfentrazone + diurondoses in pre-emergence. The experiment was conducted in Londrina PR, in an oxisol. The treatments were doses ofsulfentrazone + diuron (62 + 123, 123 + 245, 184 + 368, 245 + 490, 306 + 613 e 368 + 735 g a.i.ha-1). In addition to these treatments, isolated applications of sulfentrazone (245 g a.i.ha-1) and diuron (490 g a.i.ha-1) were evaluated, as well as a weedyand aweed-freecontrol. For the weed control, the wild radish(Raphanus raphanistrum)and itchgrass (Rottboellia cochinchinensis) emergencewere evaluated. In soybean, photosystem II (PSII)activity, stand, plant height, number of podsper plant, height of insertion of the first pod, number of nodes per plant, mass of a thousand grains and grain yield were evaluated. Results show that the mixture of sulfentrazone + diuron resulted in less than 50% wild radish control, regardless of the dose evaluated. However, for itchgrass, the control was greater than 90% in doses from 123 g sulfentrazone + 245g diurona.i.ha-1. Regarding thesoybeancrop, in doses from 245 g sulfentrazone + 490 g diuron a.i. ha-1 there was a greater number of pods per plant and higher grain yield, of about3,000 kg ha-1. The application of herbicides led to a transient inhibition of PSIIactivity, which did not result in a reduction in crop grainyield, indicating plant recovery. Therefore, the mixture of sulfentrazone + diuron is a good alternative for the weed management in pre-emergence of soybean, and should be positioned according to the history of fieldinfestation.(AU)

A utilização de herbicidas pré-emergentes é uma ferramenta importante no manejo de plantas daninhas. Recentemente foi lançada no mercado a mistura de sulfentrazone + diuron. O objetivo deste trabalho foi avaliar o controle de plantas daninhas e o desempenho agronômico da soja em resposta à aplicação de doses de sulfentrazone + diuron em pré-emergência. O experimento foi conduzido em Londrina PR, em latossolo. Os tratamentos avaliados foram doses de sulfentrazone + diuron (62 + 123, 123 + 245, 184 + 368, 245 + 490, 306 + 613 e 368 + 735 g i.a.ha-1).Além desses tratamentos, foram avaliadas as aplicações isoladas de sulfentrazone (245 g i.a.ha-1) e de diuron (490 g i.a.ha-1), além de uma testemunha infestada e outra capinada. Foram avaliados os controles de nabiça (Raphanus raphanistrum) e de capim-camalote(Rottboellia cochinchinensis). Na soja, foram avaliados a atividade do fotossistema II (FSII), a emergência, estatura de plantas, número de legumes por planta, altura de inserção do primeiro legume, número de nós por planta, massa de mil grãos e produtividade de grãos. A mistura desulfentrazone + diuron apresentoucontrole de nabiçainferior a 50%, independentemente da dose avaliada. No entanto, para o capim-camalote, o controle foi satisfatório, sendo superior a 90% em doses a partir de 123g sulfentrazone+ 245 g diuroni.a.ha-1. Em relação àsoja, em doses a partir de 245g sulfentrazone+ 490 gdiuroni.a.ha-1houve maior número de legumes por planta e maior produtividade de grãos, de aproximadamente 3.000 kg ha-1. A aplicação dos herbicidas levou a uma inibição transitória da atividade doFSII, a qual não resultou em redução da produtividade da cultura, indicando a recuperação das plantas. Portanto, a mistura de sulfentrazone + diuron é uma boa alternativa para o manejo de plantas daninhas em pré-emergência da cultura da soja, devendo ser posicionadade acordo com o histórico de infestação da lavoura.(AU)

Weed control and soybean agronomic performance in response to application of sulfentrazone + diuron in pre-emergence / Controle de plantas daninhas e desempenho agronômico de soja em resposta à aplicação de sulfentrazone + diuron em pré-emergência

The use of pre-emergent herbicides is an important tool in weedmanagement. Sulfentrazone + diuron mixture was recently launched ontothe market. The objectiveof this work was to evaluate the weed control and the agronomic performance of soybean in response to application of sulfentrazone + diurondoses in pre-emergence. The experiment was conducted in Londrina PR, in an oxisol. The treatments were doses ofsulfentrazone + diuron (62 + 123, 123 + 245, 184 + 368, 245 + 490, 306 + 613 e 368 + 735 g a.i.ha-1). In addition to these treatments, isolated applications of sulfentrazone (245 g a.i.ha-1) and diuron (490 g a.i.ha-1) were evaluated, as well as a weedyand aweed-freecontrol. For the weed control, the wild radish(Raphanus raphanistrum)and itchgrass (Rottboellia cochinchinensis) emergencewere evaluated. In soybean, photosystem II (PSII)activity, stand, plant height, number of podsper plant, height of insertion of the first pod, number of nodes per plant, mass of a thousand grains and grain yield were evaluated. Results show that the mixture of sulfentrazone + diuron resulted in less than 50% wild radish control, regardless of the dose evaluated. However, for itchgrass, the control was greater than 90% in doses from 123 g sulfentrazone + 245g diurona.i.ha-1. Regarding thesoybeancrop, in doses from 245 g sulfentrazone + 490 g diuron a.i. ha-1 there was a greater number of pods per plant and higher grain yield, of about3,000 kg ha-1. The application of herbicides led to a transient inhibition of PSIIactivity, which did not result in a reduction in crop grainyield, indicating plant recovery. Therefore, the mixture of sulfentrazone + diuron is a good alternative for the weed management in pre-emergence of soybean, and should be positioned according to the history of fieldinfestation.

A utilização de herbicidas pré-emergentes é uma ferramenta importante no manejo de plantas daninhas. Recentemente foi lançada no mercado a mistura de sulfentrazone + diuron. O objetivo deste trabalho foi avaliar o controle de plantas daninhas e o desempenho agronômico da soja em resposta à aplicação de doses de sulfentrazone + diuron em pré-emergência. O experimento foi conduzido em Londrina PR, em latossolo. Os tratamentos avaliados foram doses de sulfentrazone + diuron (62 + 123, 123 + 245, 184 + 368, 245 + 490, 306 + 613 e 368 + 735 g i.a.ha-1).Além desses tratamentos, foram avaliadas as aplicações isoladas de sulfentrazone (245 g i.a.ha-1) e de diuron (490 g i.a.ha-1), além de uma testemunha infestada e outra capinada. Foram avaliados os controles de nabiça (Raphanus raphanistrum) e de capim-camalote(Rottboellia cochinchinensis). Na soja, foram avaliados a atividade do fotossistema II (FSII), a emergência, estatura de plantas, número de legumes por planta, altura de inserção do primeiro legume, número de nós por planta, massa de mil grãos e produtividade de grãos. A mistura desulfentrazone + diuron apresentoucontrole de nabiçainferior a 50%, independentemente da dose avaliada. No entanto, para o capim-camalote, o controle foi satisfatório, sendo superior a 90% em doses a partir de 123g sulfentrazone+ 245 g diuroni.a.ha-1. Em relação àsoja, em doses a partir de 245g sulfentrazone+ 490 gdiuroni.a.ha-1houve maior número de legumes por planta e maior produtividade de grãos, de aproximadamente 3.000 kg ha-1. A aplicação dos herbicidas levou a uma inibição transitória da atividade doFSII, a qual não resultou em redução da produtividade da cultura, indicando a recuperação das plantas. Portanto, a mistura de sulfentrazone + diuron é uma boa alternativa para o manejo de plantas daninhas em pré-emergência da cultura da soja, devendo ser posicionadade acordo com o histórico de infestação da lavoura.

Use of Modified Phenolic Thyme Extracts (Thymus vulgaris) with Reduced Polyphenol Oxidase Substrates as Anthocyanin Color and Stability Enhancing Agents.

Residual enzymatic activity in certain foods, particularly of polyphenoloxidase (PPO), is responsible for the majority of anthocyanin degradation in food systems, causing also parallel losses of other relevant nutrients. The present work explored the feasibility of modifying phenolic profiles of thyme extracts, by use of chromatographic resins, to obtain phenolic extracts capable of enhancing anthocyanin colour and stability in the presence of PPO activity. Results indicated that pretreatment of thyme extracts with strong-anion exchange resins (SAE) enhanced their copigmentation abilities with strawberry juice anthocyanins. Phenolic chromatographic profiles, by HPLC-PDA, also demonstrated that thyme extracts subjected to SAE treatments had significantly lower concentrations of certain phenolic compounds, but extracts retained their colour enhancing and anthocyanin stabilization capacities though copigmentation. Additional testing also indicated that SAE modified extract had a lower ability (73% decrease) to serve as PPO substrate, when compared to the unmodified extract. Phenolic profile modification process, reported herein, could be potentially used to manufacture modified anthocyanin-copigmentation food and cosmetic additives for colour-stabilizing applications with lower secondary degradation reactions in matrixes that contain PPO activity.

Enhanced oral bioavailability of nevirapine within micellar nanocarriers compared with Viramune(®).

In this work, Nevirapine (NVP) was encapsulated within three derivatives of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) block copolymers (Tetronic(®) 904, 1107 and Pluronic(®) F127) with and without the addition of three pharmaceutical cosolvents (glycerin, propylene glycol and polyethylene glycol 400) over a wider range of concentrations (0-40% v/v). Also, we evaluated the effect of addition of the cosolvents on the micellar size as determined by dynamic light scattering (DLS) measurements and transmission electron microscopy (TEM). The solubilization capacity of the systems was investigated by UV-spectrophotometry (282nm) and the systems stability was evaluated for 1 month at 25°C. Finally, oral bioavailability of the NVP-loaded micellar systems (2mg/mL) was assessed in male Wistar rats (8mg/kg) and compared with a pediatric commercially available formulation (Viramune(®)). The present study demonstrates that PEO-PPO-PEO polymeric micelles were able to enhance apparent aqueous solubility of NVP with the addition of cosolvents. Moreover, micellar nanocarriers significantly (p<0.05) improved the oral bioavailability of the drug versus Viramune(®). Overall results support the suitability of the strategy toward the development of an optimized NVP aqueous formulation to prevent HIV/AIDS mother-to-child transmission.

AISLAMIENTO Y CARACTERIZACIÓN DE UNA POLIFENOLOXIDASA RELACIONADA CON LA TOLERANCIA DEL CLAVEL (Dianthus caryophyllus) A Fusarium oxysporum f. sp. dianthi raza 2 / Isolation and Characterization of a Polyphenoloxidase Related with Carnation (Dianthus caryophyllus) Resistence to Fusarium oxysporum f. sp. dianthi raza 2

Se purificó y caracterizó bioquímicamente la enzima polifenoloxidasa (PFO) inducida en tallos de clavel (Dianthus caryophyllus L) de variedad tolerante por inoculación con el patógeno Fusarium oxysporum f. sp. dianthi raza 2 (FOD2), causante del marchitamiento vascular. La purificación se logró a través de procesos sucesivos de cromatografía en columna de intercambio iónico, de interacción hidrofóbica y de exclusión molecular. A través de éstos se logró un factor de purificación de 314 veces con respecto al extracto inicial. La proteína purificada mostró actividad PFO y una única banda en SDSPAGE correspondiente a 40 kDa. Usando catecol como sustrato, se determinó su temperatura óptima en 45 ºC y su pH óptimo en 7,5. La enzima presentó una cinética tipo MichaelisMenten con un valor Km de 249 mM y Vmáx 322 U/min. El punto isoeléctrico (PI= 5,0) permitió establecer que se trata de una proteína de tipo ácido. Con la enzima purificada se realizaron ensayos in vitro de actividad fungitóxica, usando sus productos de reacción enfrentados al hongo FOD2, encontrándose una actividad inhibitoria importante de cerca del 57% a las 24 horas, lo que permite postular su papel en los mecanismos de defensa del clavel contra este patógeno vascular.

Polyphenoloxidase (PPO) was isolated and purified from stems of carnation (Dianthus caryophyllus L) after the inoculation with Fusarium oxysporum f. sp. dianthi race 2 (FOD2) of a tolerant variety. Purification was performed using column chromatography of ionic exchange, hydrophobic interaction and molecular exclusion. With these successive steps, a factor of 314 fold purification was achieved. The SDSPAGE analysis showed one band of 40 kDa. The purified enzyme was partially characterized using cathecol as substrate and determined the optimal temperature as 45ºC and 7.5 as optimal pH. The enzyme showed a MichaelisMenten kinetic with Km of 249 mM and Vmax 322 U/min. The IP was 5.0 indicating the acidic character of this protein. Using the purified enzyme and its reaction products, in vitro fungitoxic assays were realized indicating an important inhibitory activity against FOD2 of 57% at 24 hours. Then, it is possible to postulate that this enzyme is activated as a part of the defense mechanisms in this interaction model.