Spatial dynamics of a population with stage-dependent diffusion

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biotransformed soybean extract induces cell death of estrogen-dependent breast cancer cells by modulation of apoptotic proteins

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

Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors

Hepatitis C virus (HCV) envelope protein 2 (E2) is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr) and CD81 in human umbilical vein endothelial cells (HUVEC) and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not) on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS) and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients.

Apoptosis and reduced microvascular density of the lamina propria during tooth eruption in rats

During tooth eruption, structural and functional changes must occur in the lamina propria to establish the eruptive pathway. In this study, we evaluate the structural changes that occur during lamina propria degradation and focus these efforts on apoptosis and microvascular density. Fragments of maxilla containing the first molars from 9-, 11-, 13- and 16-day-old rats were fixed, decalcified and embedded in paraffin. The immunohistochemical detection of vascular endothelial growth factor (VEGF), caspase-3 and MAC387 (macrophage marker), and the TUNEL method were applied to the histological molar sections. The numerical density of TUNEL-positive cells and VEGF-positive blood vessel profiles were also obtained. Data were statistically evaluated using a one-way anova with the post-hoc Kruskal-Wallis or Tukey test and a significance level of P ≤ 0.05. Fragments of maxilla were embedded in Araldite for analysis under transmission electron microscopy (TEM). TUNEL-positive structures, fibroblasts with strongly basophilic nuclei and macrophages were observed in the lamina propria at all ages. Using TEM, we identified processes of fibroblasts or macrophages surrounding partially apoptotic cells. We found a high number of apoptotic cells in 11-, 13- and 16-day-old rats. We observed VEGF-positive blood vessel profiles at all ages, but a significant decrease in the numerical density was found in 13- and 16-day-old rats compared with 9-day-old rats. Therefore, the establishment of the eruptive pathway during the mucosal penetration stage depends on cell death by apoptosis, the phagocytic activity of fibroblasts and macrophages, and a decrease in the microvasculature due to vascular cell death. These data point to the importance of vascular rearrangement and vascular neoformation during tooth eruption and the development of oral mucosa.

Using landscape metrics to analyze micro-scale soil erosion processes

Methods of recording soil erosion using photographs exist but they are not commonly considered in scientific studies. Digital images may hold an expressive amount of information that can be extracted quickly in different manners. The investigation of several metrics that were initially developed for landscape ecology analysis constitutes one method. In this study we applied a method of landscape metrics to quantify the spatial configuration of surface micro-topography and erosion-related features, in order to generate a possible complementary tool for environmental management. In a 3.7 m wide and 9.7 m long soil box used during a rainfall simulation study, digital images were systematically acquired in four instances: (a) when the soil was dry; (b) after a short duration rain for initial wetting; (c) after the first erosive rain; and (d) after the 2nd erosive rain. Thirteen locations were established in the box and digital photos were taken at these locations with the camera positioned at the same orthogonal distance from the soil surface under the same ambient light intensity. Digital photos were converted into bimodal images and seven landscape metrics were analyzed: percentage of land, number of patches, density of patches, largest patch index, edge density, shape index, and fractal dimension. Digital images were an appropriate tool because they can generate data very quickly. The landscape metrics were sensitive to changes in soil surface micro-morphology especially after the 1st erosive rain event, indicating significant erosional feature development between the initial wetting and first erosive rainfall. The method is considered suitable for spatial patterns of soil micro-topography evolution from rainfall events that bear similarity to landscape scale pattern evolution from eco-hydrological processes. Although much more study is needed for calibrating the landscape metrics at the micro-scale, this study is a step forward in demonstrating the advantages of the method.

Taking account of dependent species in management of the Southern Ocean krill fishery: estimating crabeater seal abundance off east Antarctica

1. The crabeater seal Lobodon carcinophaga is considered to be a key species in the krill-based food web of the Southern Ocean. Reliable estimates of the abundance of this species are necessary to allow the development of multispecies, predator–prey models as a basis for management of the krill fishery in the Southern Ocean. 2. A survey of crabeater seal abundance was undertaken in 1500 000 km2 of pack-ice off east Antarctica between longitudes 64–150° E during the austral summer of 1999/2000. Sighting surveys, using double observer line transect methods, were conducted from an icebreaker and two helicopters to estimate the density of seals hauled out on the ice in survey strips. Satellite-linked dive recorders were deployed on a sample of seals to estimate the probability of seals being hauled out on the ice at the times of day when sighting surveys were conducted. Model-based inference, involving fitting a density surface, was used to infer densities in the entire survey region from estimates in the surveyed areas. 3. Crabeater seal abundance was estimated to be between 0.7 and 1.4 million animals (with 95% confidence), with the most likely estimate slightly less than 1 million. 4. Synthesis and applications. The estimation of crabeater seal abundance in Convention for the Conservation of Antarctic Marine Living Resources (CCAMLR) management areas off east Antarctic where krill biomass has also been estimated recently provides the data necessary to begin extending from single-species to multispecies management of the krill fishery. Incorporation of all major sources of uncertainty allows a precautionary interpretation of crabeater abundance and demand for krill in keeping with CCAMLR’s precautionary approach to management. While this study focuses on the crabeater seal and management of living resources in the Southern Ocean, it has also led to technical and theoretical developments in survey methodology that have widespread potential application in ecological and resource management studies, and will contribute to a more fundamental understanding of the structure and function of the Southern Ocean ecosystem.

INVESTIGATION INTO CURRENT EFFICIENCY FOR PULSE ELECTROCHEMICAL MACHINING OF NICKEL ALLOY

INVESTIGATION INTO CURRENT EFFICIENCY FOR PULSE ELECTROCHEMICAL MACHINING OF NICKEL ALLOY Yu Zhang, M.S. University of Nebraska, 2010 Adviser: Kamlakar P. Rajurkar Electrochemical machining (ECM) is a nontraditional manufacturing process that can machine difficult-to-cut materials. In ECM, material is removed by controlled electrochemical dissolution of an anodic workpiece in an electrochemical cell. ECM has extensive applications in automotive, petroleum, aerospace, textile, medical, and electronics industries. Improving current efficiency is a challenging task for any electro-physical or electrochemical machining processes. The current efficiency is defined as the ratio of the observed amount of metal dissolved to the theoretical amount predicted from Faraday’s law, for the same specified conditions of electrochemical equivalent, current, etc [1]. In macro ECM, electrolyte conductivity greatly influences the current efficiency of the process. Since there is a certain limit to enhance the conductivity of the electrolyte, a process innovation is needed for further improvement in current efficiency in ECM. Pulse electrochemical machining (PECM) is one such approach in which the electrolyte conductivity is improved by electrolyte flushing in pulse off-time. The aim of this research is to study the influence of major factors on current efficiency in a pulse electrochemical machining process in macro scale and to develop a linear regression model for predicting current efficiency of the process. An in-house designed electrochemical cell was used for machining nickel alloy (ASTM B435) by PECM. The effects of current density, type of electrolyte, and electrolyte flow rate, on current efficiency under different experimental conditions were studied. Results indicated that current efficiency is dependent on electrolyte, electrolyte flow rate, and current density. Linear regression models of current efficiency were compared with twenty new data points graphically and quantitatively. Models developed were close enough to the actual results to be reliable. In addition, an attempt has been made in this work to consider those factors in PECM that have not been investigated in earlier works. This was done by simulating the process by using COMSOL software. However, it was found that the results from this attempt were not substantially different from the earlier reported studies.

Mangrove Forests and Sedimentary Processes on the South of Coast of Sao Paulo State (Brazil)

Mangrove structure and distribution is conditioned by geomorphic processes. This paper describes the response of mangroves to sedimentary processes at the Cananeia-Iguape Coastal System on the south coast of Sao Paulo State (Brazil), between latitudes 24 degrees 40`S and 25 degrees 20`S. Within six study areas 41 plots were established along 14 transects. Plot size varied according to stem density from 2mx2m to 20mx20m. Here mangroves are strongly coupled to sedimentary processes, forming discrete architectural elements within particular depositional environments or topographic settings. These sedimentary structures and progradation environments are colonized by Laguncularia racemosa, associated with the smooth cordgrass Spartina alterniflora. Rhizophora mangle occurs typically near creeklets where tidal flooding is more frequent. Where tidal influence is restricted Avicennia schaueriana becomes dominant. Erosive margins are dominated by A. schaueriana or R. mangle. Single linkage cluster analysis yields three groups (A, B and C), with high levels of similarity, providing support to the classification of the data into two broad landform categories: depositional and erosive. Group A includes plots with the least structural development (nominal stem diameter d(n) between 1.05 and 4.61cm). Group B is composed of stems of intermediate diameter (4.99 cm <= d(n) <= 5.63cm). Group C plots have the largest structural development (5.50 cm <= d(n) <= 11.10cm). The structure of mangroves (dominance and structural development) reflects responses to geomorphology and habitat change.

Homology in a context dependent predatory behavior in spiders (Araneae)

Stereotyped behaviors have been routinely used as characters for phylogeny inference, but the same cannot be said of the plastic aspects of performance, which routinely are taken as a result of ecological processes. In this paper we examine the evolution of one of these plastic behavioral phenotypes, thus fostering a bridge between ecological and evolutionary processes. Foraging behavior in spiders is context dependent in many aspects, since it varies with prey type and size, spider nutritional and developmental state, previous experience and, in webweavers, is dependent on the structure of the web. Reeling is a predatory tactic typical of cobweb weavers (Theridiidae), in which the spider moves the prey toward her by pulling the capture thread (gumfoot) to which it is adhered. Predatory reeling is dependent on the gumfoot for its expression, and has not been previously reported in orbweavers. In order to investigate the evolution of this web dependent behavior, we built artificial, pseudogumfoot lines in orbwebs and registered parameters of the predatory tactics in this modified web. Aspects of the predatory tactics of 240 individuals (12 species in 4 families) were measured, and the resulting data were optimized on the phylogeny of Orbiculariae. All species perform predatory reeling with the pseudogumfoot lines. Thus, predatory reeling is homologous for the whole Orbiculariae group. In nature, holes made by insects in ecribellate orbs produce pseudogumfoot lines (similar to out experimentally modified webs), and thus reeling occurred naturally in ecribellates. Nevertheless, outside lab conditions, predatory reeling does not occur among cribellate orbweavers, so that this behavior could not have been selected for in the cribellate ancester of orbweavers. Cribellate spiders are flexible enough as to present novel and adaptive predatory responses (reeling) even when exposed for the first time to conditions outside their usual environment. Thus, the evolution of reeling suggests and alternative mechanism for the production of evolutionary novelties; that is, the exploration of unusual ecological conditions and of the regular effects these abnormal conditions have on phenotype expression.

mu(1)- and 5-HT1-dependent mechanisms in the anterior pretectal nucleus mediate the antinociceptive effects of retrosplenial cortex stimulation in rats

Aim: This study examines if injection of cobalt chloride (CoCl2) or antagonists of muscarinic cholinergic (atropine), mu(1)-opioid (naloxonazine) or 5-HT1 serotonergic (methiothepin) receptors into the dorsal or ventral portions of the anterior pretectal nucleus (APtN) alters the antinociceptive effects of stimulating the retrosplenial cortex (RSC) in rats. Main method: Changes in the nociceptive threshold were evaluated using the tail flick or incision pain tests in rats that were electrically stimulated at the RSC after the injection of saline, CoCl2 (1 mM, 0.10 mu L) or antagonists into the dorsal or ventral APtN. Key findings: The injection of CoCl2, naloxonazine (5 mu g/0.10 mu L) or methiothepin (3 mu g/0.10 mu L) into the dorsal APtN reduced the stimulation-produced antinociception from the RSC in the rat tail flick test. Reduction of incision pain was observed following stimulation of the RSC after the injection of the same substances into the ventral APtN. The injection of atropine (10 ng/0.10 mu L) or ketanserine (5 mu g/0.10 mu L) into the dorsal or ventral APtN was ineffective against the antinociception resulting from RSC stimulation. Significance: mu(1)-opioid- and 5-HT1-expressing neurons and cell processes in dorsal and ventral APtN are both implicated in the mediation of stimulation-produced antinociception from the RSC in the rat tail flick and incision pain tests, respectively. (c) 2012 Elsevier Inc. All rights reserved.

Effect of pulse repetition rate and number of pulses in the analysis of polypropylene and high density polyethylene by nanosecond infrared laser induced breakdown spectroscopy

Pulse repetition rates and the number of laser pulses are among the most important parameters that do affect the analysis of solid materials by laser induced breakdown spectroscopy, and the knowledge of their effects is of fundamental importance for suggesting analytical strategies when dealing with laser ablation processes of polymers. In this contribution, the influence of these parameters in the ablated mass and in the features of craters was evaluated in polypropylene and high density polyethylene plates containing pigment-based PbCrO4. Surface characterization and craters profile were carried out by perfilometry and scanning electron microscopy. Area, volume and profile of craters were obtained using Taylor Map software. A laser induced breakdown spectroscopy system consisted of a Q-Switched Nd:YAG laser (1064 nm, 5 ns) and an Echelle spectrometer equipped with ICCD detector were used. The evaluated operating conditions consisted of 10, 25 and 50 laser pulses at 1, 5 and 10 Hz, 250 mJ/pulse (85 J cm(-2)), 2 mu s delay time and 6 mu s integration time gate. Differences in the topographical features among craters of both polymers were observed. The decrease in the repetition rate resulted in irregular craters and formation of edges, especially in polypropylene sample. The differences in the topographical features and ablated masses were attributed to the influence of the degree of crystallinity, crystalline melting temperature and glass transition temperature in the ablation process of the high density polyethylene and polypropylene. It was also observed that the intensities of chromium and lead emission signals obtained at 10 Hz were two times higher than at 5 Hz by keeping the number of laser pulses constant. (C) 2011 Elsevier B. V. All rights reserved.

Behavior of the thermal diffusivity of native and oxidized human low-density lipoprotein solutions studied by the Z-scan technique

Modifications in low-density lipoprotein (LDL) have emerged as a major pathogenic factor of atherosclerosis, which is the main cause of morbidity and mortality in the western world. Measurements of the heat diffusivity of human LDL solutions in their native and in vitro oxidized states are presented by using the Z-Scan (ZS) technique. Other complementary techniques were used to obtain the physical parameters necessary to interpret the optical results, e. g., pycnometry, refractometry, calorimetry, and spectrophotometry, and to understand the oxidation phase of LDL particles. To determine the sample's thermal diffusivity using the thermal lens model, an iterative one-parameter fitting method is proposed which takes into account several characteristic ZS time-dependent and the position-dependent transmittance measurements. Results show that the thermal diffusivity increases as a function of the LDL oxidation degree, which can be explained by the increase of the hydroperoxides production due to the oxidation process. The oxidation products go from one LDL to another, disseminating the oxidation process and caring the heat across the sample. This phenomenon leads to a quick thermal homogenization of the sample, avoiding the formation of the thermal lens in highly oxidized LDL solutions. (C) 2012 Society of Photo-Optical Instrumentation Engineers (SPIE). [DOI: 10.1117/1.JBO.17.10.105003]

Laminated Tubular-channel Ferroelectret Systems from Low-density Polyethylene Films and from Fluoroethylene-propylene Copolymer Films - A Comparison

A template-based lamination technique for the manufacture of ferroelectrets from uniform electret films was recently reported. In the present work, this technique is used to prepare similar ferroelectret structures from low-density polyethylene (LDPE) films and from fluoro-ethylene-propylene (FEP) copolymer films. A comparative analysis of the pressure-, temperature-, and frequency-dependent piezoelectric properties has been performed on the two ferroelectret systems. It is observed that the FEP ferroelectrets exhibit better piezoelectric responses and are thermally more stable. The difference between the piezoelectric d(33) coefficients of the two ferroelectret systems is partially explained here by their different elastic moduli. The anti-resonance peaks of both structures have been investigated by means of dielectric resonance spectroscopy and electroacoustic sound-pressure measurements. A difference of more than 10 kHz is observed between the anti-resonance frequencies of the two ferroelectret systems.

Black hole formation with an interacting vacuum energy density

We discuss the gravitational collapse of a spherically symmetric massive core of a star in which the fluid component is interacting with a growing vacuum energy density. The influence of the variable vacuum in the collapsing core is quantified by a phenomenological beta parameter as predicted by dimensional arguments and the renormalization group approach. For all reasonable values of this free parameter, we find that the vacuum energy density increases the collapsing time, but it cannot prevent the formation of a singular point. However, the nature of the singularity depends on the value of beta. In the radiation case, a trapped surface is formed for beta <= 1/2, whereas for beta >= 1/2, a naked singularity is developed. In general, the critical value is beta = 1-2/3(1 + omega) where omega is the parameter describing the equation of state of the fluid component.

Platelet-activating factor receptor (PAF-R)-dependent pathways control tumour growth and tumour response to chemotherapy

Abstract Background Phagocytosis of apoptotic cells by macrophages induces a suppressor phenotype. Previous data from our group suggested that this occurs via Platelet-activating factor receptor (PAF-R)-mediated pathways. In the present study, we investigated the impact of apoptotic cell inoculation or induction by a chemotherapeutic agent (dacarbazine, DTIC) on tumour growth, microenvironmental parameters and survival, and the effect of treatment with a PAF-R antagonist (WEB2170). These studies were performed in murine tumours: Ehrlich Ascitis Tumour (EAT) and B16F10 melanoma. Methods Tumour growth was assessed by direct counting of EAT cells in the ascitis or by measuring the volume of the solid tumour. Parameters of the tumour microenvironment, such as the frequency of cells expressing cyclo-oxygenase-2 (COX-2), caspase-3 and galectin-3, and microvascular density, were determined by immunohistochemistry. Levels of vascular endothelium growth factor (VEGF) and prostaglandin E2 (PGE2) were determined by ELISA, and levels of nitric oxide (NO) by Griess reaction. PAF-R expression was analysed by immunohistochemistry and flow cytometry. Results Inoculation of apoptotic cells before EAT implantation stimulated tumour growth. This effect was reversed by in vivo pre-treatment with WEB2170. This treatment also reduced tumour growth and modified the microenvironment by reducing PGE2, VEGF and NO production. In B16F10 melanoma, WEB2170 alone or in association with DTIC significantly reduced tumour volume. Survival of the tumour-bearing mice was not affected by WEB2170 treatment but was significantly improved by the combination of DTIC with WEB2170. Tumour microenvironment elements were among the targets of the combination therapy since the relative frequency of COX-2 and galectin-3 positive cells and the microvascular density within the tumour mass were significantly reduced by treatment with WEB2170 or DTIC alone or in combination. Antibodies to PAF-R stained the cells from inside the tumour, but not the tumour cells grown in vitro. At the tissue level, a few cells (probably macrophages) stained positively with antibodies to PAF-R. Conclusions We suggest that PAF-R-dependent pathways are activated during experimental tumour growth, modifying the microenvironment and the phenotype of the tumour macrophages in such a way as to favour tumour growth. Combination therapy with a PAF-R antagonist and a chemotherapeutic drug may represent a new and promising strategy for the treatment of some tumours.