Influência do 1-MCP na conservação pós-colheita de rosas cv. Vega

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

Produção, qualidade dos frutos e estado nutricional da laranja valência sob fertirrigação e adubação convencional

O objetivo deste trabalho foi comparar os efeitos de doses de nitrogênio, fósforo e potássio aplicadas via água de irrigação em comparação à aplicação convencional de fertilizantes, com irrigação e sem irrigação. O experimento foi conduzido em um pomar de laranja 'Valência' (Citrus sinensis Osbeck) localizado em Pratânia - SP, durante três anos consecutivos, e analisados os dados do último ano de observação. As diferentes formas de aplicação dos fertilizantes e a redução das doses fornecidas via água de irrigação não resultaram em diferenças significativas sobre o número médio de frutos e produtividade. Também não foram constatadas diferenças estatísticas dos tratamentos sobre a qualidade do suco e o estado nutricional das plantas, com exceção do teor de fósforo.

Effect of oxytocin on transepithelial transport of water and Na+ in distinct ventral regions of frog skin (Rana catesbeiana)

Thoracic, abdominal, and pelvic fragments of ventral skin of Rana catesbeiana were analysed regarding the effect of oxytocin on: (1) transepithelial water transport; (2) short-circuit current, (3) skin conductance and electrical potential difference; (4) Na+ conductance, the electromotive force of the Nat transport mechanism, and shunt conductance; (5) short-circuit current responses to fast Na+ by K+ replacement in the outer compartment, and (6) epithelial microstructure. Unstimulated water and Na+ permeabilities were low along the ventral skin. Hydrosmotic and natriferic responses to oxytocin increased from thorax to pelvis, Unstimulated Na+ conductance was greater in pelvis than in abdomen, the other electrical parameters being essentially similar in both skin fragments. Contribution of shunt conductance to total skin conductance was higher in abdominal than in pelvic skin. Oxytocin-induced increases of total skin conductance, Na+ conductance, and shunt conductance in pelvis were significantly larger than in abdomen, An oscillatory behaviour of the short-circuit current was observed only in oxytocin-treated pelvic skins. Decrease of epithelial thickness and increase of mitochondria-rich cell number were observed from thorax to pelvis, Oxytocin-induced increases of interspaces were more conspicuous in pelvis and abdomen than in thorax.

A New Indirect Electrochemical Method for Determination of Ozone in Water Using Multiwalled Carbon Nanotubes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Water table effects on bean yield and nitrate distribution in the soil profile

In order to evaluate the bean yield under different water table levels as well as the moisture and nitrate distribution in the soil profile, a field experiment was carried out at the experimental area from the College of Agronomic Sciences - UNESP, Botucatu, SP, Brazil. Beans were grown in field lysimeters and subjected to five water table depths:30; 40; 50; 60 and 70 cm. The moisture in the soil profile was gravimetrically determined through samples obtained at 10; 20; 30; 40; 50; 60 and 70cm of depth. The water table depths of 30cm and 40cm showed the highest productivities (3,228.4 kg.ha-1 and 3,422.1 kg.ha-1, respectively), showing no statistical differences between each other. The highest productivity was related to the two most elevated water table levels (30 and 40cm), which provided the highest moisture average values on basis of volume in the soil profile (33.3 e 31%) as well as the consumptive use of water (416 and 396 mm). The nitrate content during the bean cycle at the extraction depth of 60cm has been under the safe drinking limit of 10 mg.1-1 for water table depths of 30; 40; 50 and 60cm, showing the denitrification effectiveness as a way of controlling water table from nitrate pollution. The water table handling allowed the attainment of high bean productivity levels, as well as the reduction of the nitrate level.

Water relations of cut inflorescences of Alpinia purpurata treated with seven pulsing solutions

Alpinia purpurata (Vieill) K. Schum, common name red ginger belongs to Zingiberaceae family, and is a perennial, with nice inflorescences composed of layers of bracts arranged in spirals. The utilization of keeping-quality solutions seeks to prolong vase-life and to maintain the quality of cut flowers. This research was performed to evaluate the effect of seven pulsing solutions (during 24 hours) on water relations, quality (turgor, browning and curvature) and the longevity of cut red ginger. The experiment followed complete randomized design, in factorial arrangement. The following treatments were studied: 1) distilled water (control), 2) sucrose 2% + 8-hidroxyquinoline citrate 200 ppm, 3) sucrose 2%, 4) sucrose 2% + citric acid, 5) benzyladenine 10 μM, 6) benzyladenine 10 μmol + sucrose 2%, 7) quaternary ammonia 0,5 mL/L. There was no significant (P>0.05) reduction of the relative water content (RWC) of the inflorescence bracts, among solutions, in the first period (7 days) and in the second period (14 days) and, between the two periods for all treatments. Significant differences among RWC of some treatments were recorded after 16 days of vase life. The utilization of benzyladenine, benzyladenine plus sucrose and sucrose plus citric acid, in the pulsing solution, allowed to keep the flowers quality for longer periods than the other treatments.

Water table effects on bean yield and nitrate distribution in the soil profile

In order to evaluate the bean yield under different water table levels as well as the moisture and nitrate distribution in the soil profile, a field experiment was carried out in the experimental area of the College of Agricultural Sciences - UNESP, Botucatu, SP, Brazil. Beans were grown in field lysimeters under five water table depths: 30; 40; 50; 60 and 70 cm. The moisture in the soil profile was determined gravimetrically using samples collected at 10; 20; 30; 40; 50; 60 and 70 cm deep. The water table depths of 30cm and 40cm showed the highest productivities (3,228.4kg.ha-1 and 3,422.1kg.ha-1, respectively), with no statistical differences between them. The highest productivity was related to the two highest water table levels (30 and 40cm), which provided the highest moisture average values on the basis of volume in the soil profile (33.3 e 31%) as well as the consumptive use of water (416 and 396mm). The nitrate content during the bean cycle at the extraction depth of 60cm was below the safe drinking limit of 10mg.1-1 for water table depths of 30; 40; 50 and 60cm, which shows the denitrification efficiency as a way of controlling nitrate pollution in water tables. The management of water table can lead to high levels of bean yield and to a better control of nitrate pollution in underground water.

Necessidade Hídrica E Coeficiente de Cultivo do Pimentão Fertirrigado

This study aimed to determine the crop evapotranspiration (ETc) and crop coefficient (Kc) of fertigated sweet pepper, measured over time (15, 30, 45, 60, 75, 90 and 105 days after the transplant). The experiment was conducted on a battery of 40 drainage lysimeters, arranged in a 8 x 5 matrix, in the period October/2009 to January/2010, located in the Federal Rural University of Pernambuco (UFRPE) Campus in Recife, in a completely randomized factorial scheme 5 x 2, with four replications, being five irrigation depths (80, 90, 100, 110 and 120% of ETc) and two levels of potassium (80 and 120 kg K2O ha-1) where the experimental plot was composed of three plants in each lysimeter. The Mauchly Sphericity Test indicated multivariate variance analysis for the studied variables ETc (mm d-1) and Kc, the variables presented adjustment under quadratic effect of variation on observation times, when the irrigation depths were fixed, and descending linear adjustments under the variation of irrigation depths effect, when the times of observation were fixed in each potassium dose.

Estádios de maturação de pêssegos 'Aurora-1' para o processamento mínimo

The experiment aimed to verify the effect of maturation stage on the quality of minimally processed peaches. Fruits were used in two stages of maturity, on time corresponding to the background color yellow-green, and mature, which corresponds completely to the background color yellow. The minimum process consisted of washing, sanitizing, enzyme peel, cut lengthwise and removed the stone fruits. Halves obtained were immersed in water chlorinated to 10 mg L-1 of water and left on standing to drain the excess liquid. Afterwards, it was proceeded the packaging of the halves in containers of polyethylene terephthalate (PET) and with transparent lid, and the storage at 3 ± 2 ° C and RH = 65% for 12 days, with assessments every three days. The variables evaluated were appearance, weight loss, firmness, soluble solids, titratable acidity, soluble and reduced sugars, ascorbic acid, total and soluble pectin, coloring and activity of polyphenoloxidase. The storage of minimally processed peaches 'Aurora-1' mature harvested was limited mainly by the loss of freshness and firmness, and because they have darker appearance, and lower levels of reduced sugars and ascorbic acid. Peaches 'Aurora-1', harvested at the maturity stage on time, had better quality and longer duration of their minimally processed products.

Avaliação do crescimento de plantas de milho em função de doses de potássio e estresse hídrico

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

The effect of ions and cryoprotectants upon sperm motility and fertilization success in the loach Misgurnus anguillicaudatus

The solutions commonly used to dilute or cryopreserve sperm are commonly composed of salts, buffers and cryoprotectants, which may affect gametes and subsequent fertilization success. Here, we have evaluated the effects of several cryoprotectants (methanol; MeOH, dimethyl sulfoxide; DMSO and dimethyl acetamide; DMA at concentrations of 0.25, 0.5 and 1%) and different ions (potassium, calcium and magnesium at concentrations of 1.25, 2.5, 5.0 and 10 mM) as sperm diluents upon sperm motility and fertilization success in the loach Misgurnus anguillicaudatus sperm. Our results demonstrated that DMSO (at 1%) decreased sperm motility while calcium and magnesium ions (from 2.5 mM) induced sperm aggregation and reduced sperm motility. Reduced fertilization rates were observed with potassium (from 1.25 mM), calcium (at 10 mM), magnesium (at 10 mM), DMA (at 1%), and DMSO (at 1%). We conclude that specific ions and cryoprotectants, and their relative concentrations caused effect upon loach gametes. These data are important to consider for the preparation of sperm diluents and activating solutions in order to manage gamete quality for artificial propagation. (C) 2012 Elsevier B.V. All rights reserved.

Computational analyses on the structure-function relation in ion channels

Ion channels are protein molecules, embedded in the lipid bilayer of the cell membranes. They act as powerful sensing elements switching chemicalphysical stimuli into ion-fluxes. At a glance, ion channels are water-filled pores, which can open and close in response to different stimuli (gating), and one once open select the permeating ion species (selectivity). They play a crucial role in several physiological functions, like nerve transmission, muscular contraction, and secretion. Besides, ion channels can be used in technological applications for different purpose (sensing of organic molecules, DNA sequencing). As a result, there is remarkable interest in understanding the molecular determinants of the channel functioning. Nowadays, both the functional and the structural characteristics of ion channels can be experimentally solved. The purpose of this thesis was to investigate the structure-function relation in ion channels, by computational techniques. Most of the analyses focused on the mechanisms of ion conduction, and the numerical methodologies to compute the channel conductance. The standard techniques for atomistic simulation of complex molecular systems (Molecular Dynamics) cannot be routinely used to calculate ion fluxes in membrane channels, because of the high computational resources needed. The main step forward of the PhD research activity was the development of a computational algorithm for the calculation of ion fluxes in protein channels. The algorithm - based on the electrodiffusion theory - is computational inexpensive, and was used for an extensive analysis on the molecular determinants of the channel conductance. The first record of ion-fluxes through a single protein channel dates back to 1976, and since then measuring the single channel conductance has become a standard experimental procedure. Chapter 1 introduces ion channels, and the experimental techniques used to measure the channel currents. The abundance of functional data (channel currents) does not match with an equal abundance of structural data. The bacterial potassium channel KcsA was the first selective ion channels to be experimentally solved (1998), and after KcsA the structures of four different potassium channels were revealed. These experimental data inspired a new era in ion channel modeling. Once the atomic structures of channels are known, it is possible to define mathematical models based on physical descriptions of the molecular systems. These physically based models can provide an atomic description of ion channel functioning, and predict the effect of structural changes. Chapter 2 introduces the computation methods used throughout the thesis to model ion channels functioning at the atomic level. In Chapter 3 and Chapter 4 the ion conduction through potassium channels is analyzed, by an approach based on the Poisson-Nernst-Planck electrodiffusion theory. In the electrodiffusion theory ion conduction is modeled by the drift-diffusion equations, thus describing the ion distributions by continuum functions. The numerical solver of the Poisson- Nernst-Planck equations was tested in the KcsA potassium channel (Chapter 3), and then used to analyze how the atomic structure of the intracellular vestibule of potassium channels affects the conductance (Chapter 4). As a major result, a correlation between the channel conductance and the potassium concentration in the intracellular vestibule emerged. The atomic structure of the channel modulates the potassium concentration in the vestibule, thus its conductance. This mechanism explains the phenotype of the BK potassium channels, a sub-family of potassium channels with high single channel conductance. The functional role of the intracellular vestibule is also the subject of Chapter 5, where the affinity of the potassium channels hEag1 (involved in tumour-cell proliferation) and hErg (important in the cardiac cycle) for several pharmaceutical drugs was compared. Both experimental measurements and molecular modeling were used in order to identify differences in the blocking mechanism of the two channels, which could be exploited in the synthesis of selective blockers. The experimental data pointed out the different role of residue mutations in the blockage of hEag1 and hErg, and the molecular modeling provided a possible explanation based on different binding sites in the intracellular vestibule. Modeling ion channels at the molecular levels relates the functioning of a channel to its atomic structure (Chapters 3-5), and can also be useful to predict the structure of ion channels (Chapter 6-7). In Chapter 6 the structure of the KcsA potassium channel depleted from potassium ions is analyzed by molecular dynamics simulations. Recently, a surprisingly high osmotic permeability of the KcsA channel was experimentally measured. All the available crystallographic structure of KcsA refers to a channel occupied by potassium ions. To conduct water molecules potassium ions must be expelled from KcsA. The structure of the potassium-depleted KcsA channel and the mechanism of water permeation are still unknown, and have been investigated by numerical simulations. Molecular dynamics of KcsA identified a possible atomic structure of the potassium-depleted KcsA channel, and a mechanism for water permeation. The depletion from potassium ions is an extreme situation for potassium channels, unlikely in physiological conditions. However, the simulation of such an extreme condition could help to identify the structural conformations, so the functional states, accessible to potassium ion channels. The last chapter of the thesis deals with the atomic structure of the !- Hemolysin channel. !-Hemolysin is the major determinant of the Staphylococcus Aureus toxicity, and is also the prototype channel for a possible usage in technological applications. The atomic structure of !- Hemolysin was revealed by X-Ray crystallography, but several experimental evidences suggest the presence of an alternative atomic structure. This alternative structure was predicted, combining experimental measurements of single channel currents and numerical simulations. This thesis is organized in two parts, in the first part an overview on ion channels and on the numerical methods adopted throughout the thesis is provided, while the second part describes the research projects tackled in the course of the PhD programme. The aim of the research activity was to relate the functional characteristics of ion channels to their atomic structure. In presenting the different research projects, the role of numerical simulations to analyze the structure-function relation in ion channels is highlighted.

Frog oocytes to unveil the structure and supramolecular organization of human transport proteins

Structural analyses of heterologously expressed mammalian membrane proteins remain a great challenge given that microgram to milligram amounts of correctly folded and highly purified proteins are required. Here, we present a novel method for the expression and affinity purification of recombinant mammalian and in particular human transport proteins in Xenopus laevis frog oocytes. The method was validated for four human and one murine transporter. Negative stain transmission electron microscopy (TEM) and single particle analysis (SPA) of two of these transporters, i.e., the potassium-chloride cotransporter 4 (KCC4) and the aquaporin-1 (AQP1) water channel, revealed the expected quaternary structures within homogeneous preparations, and thus correct protein folding and assembly. This is the first time a cation-chloride cotransporter (SLC12) family member is isolated, and its shape, dimensions, low-resolution structure and oligomeric state determined by TEM, i.e., by a direct method. Finally, we were able to grow 2D crystals of human AQP1. The ability of AQP1 to crystallize was a strong indicator for the structural integrity of the purified recombinant protein. This approach will open the way for the structure determination of many human membrane transporters taking full advantage of the Xenopus laevis oocyte expression system that generally yields robust functional expression.