Extensive structural change of the envelope protein of dengue virus induced by a tuned ionic strength: conformational and energetic analyses

The Dengue has become a global public health threat, with over 100 million infections annually; to date there is no specific vaccine or any antiviral drug. The structures of the envelope (E) proteins of the four known serotype of the dengue virus (DENV) are already known, but there are insufficient molecular details of their structural behavior in solution in the distinct environmental conditions in which the DENVs are submitted, from the digestive tract of the mosquito up to its replication inside the host cell. Such detailed knowledge becomes important because of the multifunctional character of the E protein: it mediates the early events in cell entry, via receptor endocytosis and, as a class II protein, participates determinately in the process of membrane fusion. The proposed infection mechanism asserts that once in the endosome, at low pH, the E homodimers dissociate and insert into the endosomal lipid membrane, after an extensive conformational change, mainly on the relative arrangement of its three domains. In this work we employ all-atom explicit solvent Molecular Dynamics simulations to specify the thermodynamic conditions in that the E proteins are induced to experience extensive structural changes, such as during the process of reducing pH. We study the structural behavior of the E protein monomer at acid pH solution of distinct ionic strength. Extensive simulations are carried out with all the histidine residues in its full protonated form at four distinct ionic strengths. The results are analyzed in detail from structural and energetic perspectives, and the virtual protein movements are described by means of the principal component analyses. As the main result, we found that at acid pH and physiological ionic strength, the E protein suffers a major structural change; for lower or higher ionic strengths, the crystal structure is essentially maintained along of all extensive simulations. On the other hand, at basic pH, when all histidine residues are in the unprotonated form, the protein structure is very stable for ionic strengths ranging from 0 to 225 mM. Therefore, our findings support the hypothesis that the histidines constitute the hot points that induce configurational changes of E protein in acid pH, and give extra motivation to the development of new ideas for antivirus compound design.

Relationship between ozone, meteorological conditions, gas exchange and leaf injury in Nicotiana tabacum Bel-W3 in a sub-tropical region

The city of Sao Paulo is located in a subtropical region whose climate exhibits few defined seasons as well as frequent oscillations in temperature and rainfall throughout the year. In addition to interfering with physiological processes, these peculiar climatic dynamics influence the formation of O-3 and its influx into leaves, causing species used as bioindicators in temperate climates to be ineffective here. This study evaluated gas exchange variations in CO2 and H2O and leaf injuries induced by O-3 in Nicotiana tabacum Bel-W3 in relation to oscillations in environmental conditions. Plants were exposed to an O-3-polluted environment for fifteen periods of fourteen days each throughout 2008. Gas exchange and O-3 were higher during the summer and winter but were highly variable in all seasons. Severe injuries occurred during the winter and spring, with significant variation in this parameter being observed throughout the year. An analysis of biotic and abiotic variables revealed complex relationships among them, with great importance of meteorological factors in plant responses. We conclude that under unstable climatic conditions, the relationship between O-3 flux and injury is weak, and the qualitative character of biomonitoring is further confirmed. (c) 2012 Elsevier Ltd. All rights reserved.

Effect of pH, mucin and bovine serum on rifampicin permeability through Caco-2 cells

Rifampicin, a poorly soluble drug, has great importance in therapeutics as it is the main drug used to treat tuberculosis. The characterization of its permeability and the factors that influence it represent an important tool for predicting its bioavailability. Caco-2 cell monolayers were used as models of the intestinal mucosa to assess the uptake and transport of rifampicin and the effects of various experimental conditions were investigated, in order to establish the influence of these variables on rifampicin permeability. Different pHs (5.8, 6.8 and 7.4) in the apical medium, the presence or absence of mucin (3.0% w/v) in the donor site and the presence or absence of bovine serum albumin (4.0% v/v) in the receptor chamber were the evaluated conditions. The quantification of rifampicin in the apical or basolateral chambers was performed by a validated HPLC-UV method. The change in the donor chamber pH showed that permeability values were greater at pH 6.8, although this increase does not result in an alteration of the qualitative classification of rifampicin, which has high permeability. Mucin and bovine serum showed no effects on the permeability of rifampicin at the concentration tested. Overall, the current study suggests that pH, artificial mucin and bovine serum proteins have no influence on rifampicin permeability. Copyright (c) 2012 John Wiley & Sons, Ltd.

Effects of Black Leaf Streak Disease and Sigatoka Disease on fruit quality and maturation process of bananas produced in the subtropical conditions of southern Brazil

Banana fruits are harvested at the green-mature stage (pre-climacteric) in order to allow sufficient time for transport and marketing. The time between the harvest and the initiation of the natural ripening process is called green life (GL), which is closely correlated to physiological age. Sigatoka Disease (SD: also called yellow Sigatoka) and Black Leaf Streak Disease (BLSD; also called black Sigatoka) are the main foliar diseases affecting banana production. The aim of this work was to investigate the influence of these diseases on banana GL and postharvest behavior in subtropical conditions (southeastern Brazil). The results showed that both diseases shortened the banana's GL when compared to control bananas of the same physiological age. Moreover, fruits from infested plots showed higher values of CO2 (+100% for SD and +300% for BLSD) and C2H4 production (+30% for SD and +60% for BLSD) at the climacteric peak. BLSD caused 40% reduction in fruit weight. Fruits from plants with a high degree of SD or BLSD undergo an altered maturation process. (C) 2011 Elsevier Ltd. All rights reserved.

Effects of tannery sludge application on physiological and fatty acid profiles of the soil microbial community

The impact of tannery sludge application on soil microbial community and diversity is poorly understood. We studied the microbial community in an agricultural soil following two applications (2006 and 2007) of tannery sludge with annual application rates of 0.0,2.3 and 22.6 Mg ha(-1). The soil was sampled 12 and 271 days after the second (2007) application. Community structure was assessed via a phospholipid fatty acid analysis, and the physiological profile of the soil microbial community via the Biolog method. Tannery sludge application changed soil chemical properties, increasing the soil pH and electrical conductivity as well as available P and mineral N concentrations. The higher sludge application rate changed the community structure and the physiological profile of the microbial community at both sampling dates. However, there is no clear link between community structure and carbon substrate utilization. According to the Distance Based Linear Models Analysis, the fatty acids 16:0 and 117:0 together contributed 84% to the observed PLFA patterns, whereas the chemical properties available P, mineral N, and Ca, and pH together contributed 54%. At 12 days, tannery sludge application increased the average well color development from 0.46 to 0.87 after 48 h, and reduced the time elapsed before reaching the midpoint carbon substrate utilization (s) from 71 to 44 h, an effect still apparent nine months after application of the higher sludge application rate. The dominant signature fatty acids and kinetic parameters (r and s) were correlated to the concentrations of available P. Ca, mineral N, pH and EC. (c) 2012 Elsevier B.V. All rights reserved.

Relationships between gender, age, family conditions, physical and mental health, and social isolation of elderly caregivers

Background: In an aging population an increasing number of elderly caregivers will be called upon to provide care over a long period, during which time they will be burdened both by caregiving and by the physiological effects of their own aging. Among them there will be more aged male caregivers, who will probably be less prepared than women to become caregivers. The aim of this study was to investigate the relationship between caregivers' gender, age, family income, living arrangements and social support as independent variables, and depressive symptoms, comorbidities, level of frailty, grip strength, walking speed and social isolation, as dependent variables. Methods: 176 elderly people (123 women) were selected from a sample of a population-based study on frailty (n = 900), who had cared for a spouse (79.3%) and/or parents (31.4%) in the past five years (mean age = 71.8 +/- 4.86 years; mean monthly family income in minimum wages = 4.64 +/- 5.14). The study used questionnaires and self-report scales, grip strength and walking speed tests. Results: 65% of participants evaluated caregiving as being very stressful. Univariate analyses of regression showed low family income as a risk factor for depression; being female and low perceived social support as a risk for comorbidities; being 80 years of age and above for low grip strength; and being male for social isolation indicated by discontinuity of activities and social roles. In multivariate analyses of regression, poverty arose as a risk factor for depression and being female for comorbidities. Conclusions: Gender roles, age, income and social support interacted with physical and emotional health, and with the continuity of social participation of elderly caregivers. Special attention must be given to male caregivers.

Characterization of Heparin-induced Glyceraldehyde-3-phosphate Dehydrogenase Early Amyloid-like Oligomers and Their Implication in alpha-Synuclein Aggregation

Lewy bodies and Lewy neurites, neuropathological hallmarks of several neurological diseases, are mainly made of filamentous assemblies of alpha-synuclein. However, other macromolecules including Tau, ubiquitin, glyceraldehyde-3-phosphate dehydrogenase, and glycosaminoglycans are routinely found associated with these amyloid deposits. Glyceraldehyde-3-phosphate dehydrogenase is a glycolytic enzyme that can form fibrillar aggregates in the presence of acidic membranes, but its role in Parkinson disease is still unknown. In this work, the ability of heparin to trigger the amyloid aggregation of this protein at physiological conditions of pH and temperature is demonstrated by infrared and fluorescence spectroscopy, dynamic light scattering, small angle x-ray scattering, circular dichroism, and fluorescence microscopy. Aggregation proceeds through the formation of short rod-like oligomers, which elongates in one dimension. Heparan sulfate was also capable of inducing glyceraldehyde-3-phosphate dehydrogenase aggregation, but chondroitin sulfates A, B, and C together with dextran sulfate had a negligible effect. Aided with molecular docking simulations, a putative binding site on the protein is proposed providing a rational explanation for the structural specificity of heparin and heparan sulfate. Finally, it is demonstrated that in vitro the early oligomers present in the glyceraldehyde-3-phosphate dehydrogenase fibrillation pathway promote alpha-synuclein aggregation. Taking into account the toxicity of alpha-synuclein prefibrillar species, the heparin-induced glyceraldehyde-3-phosphate dehydrogenase early oligomers might come in useful as a novel therapeutic strategy in Parkinson disease and other synucleinopathies.

Norepinephrine stimulates progesterone production in highly estrogenic bovine granulosa cells cultured under serum-free, chemically defined conditions

Background: Since noradrenergic innervation was described in the ovarian follicle, the actions of the intraovarian catecholaminergic system have been the focus of a variety of studies. We aimed to determine the gonadotropin-independent effects of the catecholamine norepinephrine (NE) in the steroid hormone profile of a serum-free granulosa cell (GC) culture system in the context of follicular development and dominance. Methods: Primary bovine GCs were cultivated in a serum-free, chemically defined culture system supplemented with 0.1% polyvinyl alcohol. The culture features were assessed by hormone measurements and ultrastructural characteristics of GCs. Results: GCs produced increasing amounts of estradiol and pregnenolone for 144h and maintained ultrastructural features of healthy steroidogenic cells. Progesterone production was also detected, although it significantly increased only after 96h of culture. There was a highly significant positive correlation between estradiol and pregnenolone production in high E2-producing cultures. The effects of NE were further evaluated in a dose response study. The highest tested concentration of NE (10 (-7) M) resulted in a significant increase in progesterone production, but not in estradiol or pregnenolone production. The specificity of NE effects on progesterone productio n was further investigated by incubating GCs with propranolol (10 (-8) M), a non-selective beta-adrenergic antagonist. Conclusions: The present culture system represents a robust model to study the impact of intrafollicular factors, such as catecholamines, in ovarian steroidogenesis and follicular development. The results of noradrenergic effects in the steroidogenesis of GC have implications on physiological follicular fate and on certain pathological ovarian conditions such as cyst formation and anovulation.

Spatial variability of soil fertility and its relationship with seed physiological potential in a soybean production area

The correlation of soil fertility x seed physiological potential is very important in the area of seed technology but results published with that theme are contradictory. For this reason, this study to evaluate the correlations between soil chemical properties and physiological potential of soybean seeds. On georeferenced points, both soil and seeds were sampled for analysis of soil fertility and seed physiological potential. Data were assessed by the following analyses: descriptive statistics; Pearson's linear correlation; and geostatistics. The adjusted parameters of the semivariograms were used to produce maps of spatial distribution for each variable. Organic matter content, Mn and Cu showed significant effects on seed germination. Most variables studied presented moderate to high spatial dependence. Germination and accelerated aging of seeds, and P, Ca, Mg, Mn, Cu and Zn showed a better fit to spherical semivariogram: organic matter, pH and K had a better fit to Gaussian model; and V% and Fe showed a better fit to the linear model. The values for range of spatial dependence varied from 89.9 m for P until 651.4 m for Fe. These values should be considered when new samples are collected for assessing soil fertility in this production area.

Computerized imaging analysis of seedlings for assessment of physiological potential of wheat seeds

Nowadays, image analysis is one of the most modern tools in evaluating physiological potential of seeds. This study aimed at verifying the efficiency of the seedling imaging analysis to assess physiological potential of wheat seeds. The seeds of wheat, cultivars IAC 370 and IAC 380, each of which represented by five different lots, were stored during four months under natural environmental conditions of temperature (T) and relative humidity (RH), in municipality of Piracicaba, Stated of São Paulo, Brazil. For this, bimonthly assessments were performed to quantify moisture content and physiological potential of seeds by means of tests of: germination, first count, accelerated aging, electrical conductivity, seedling emergence, and computerized analysis of seedlings, using the Seed Vigor Imaging System (SVIS®). It has been concluded that the computerized analyses of seedling through growth indexes and vigor, using the SVIS®, is efficient to assess physiological potential of wheat seeds.

Effect of pH-induced nanopowder deagglomeration on sintering, microstructure and dielectric properties of Ba0.77Ca0.23TiO3 ceramics

Ba0.77Ca0.23TiO3 ceramics were produced in this work starting from nanopowders synthesized via a polymeric precursor method. By adjusting the pH values of the precursor solutions above 7, it was possible to prepare powders weakly aggregated and with a smaller particle size, both facts which traduced into an enhanced nanopowders' sintering process at comparatively lower temperatures. Irrespective of the initial pH value, highly-dense and second phase-free ceramics were obtained following optimal sintering parameters (temperature and time) extracted from dilatometric and density measurements. By considering these and other sintering conditions, moreover, polycrystalline materials with an average grain size varying from 0.35 to 8 mm were produced, the grain growth process involving liquid phase-assisted sintering for heat treatments achieved at 1320 °C. The study of grain size effects on the ferroelectric properties of these materials was conducted, the results being discussed in the light of previous debates, including grain size-dependent degree of tetragonal distortion in such materials, as verified in this work.

Fermentative production of ribonucleotides from whey by Kluyveromyces marxianus: effect of temperature and pH

Ribonucleotides have shown many promising applications in food and pharmaceutical industries. The aim of the present study was to produce ribonucleotides (RNA) by Kluyveromyces marxianus ATCC 8,554 utilizing cheese whey, a dairy industry waste, as a main substrate under batch fermentation conditions. The effects of temperature, pH, aeration rate, agitation and initial cellular concentration were studied simultaneously through factorial design for RNA, biomass production and lactose consumption. The maximum RNA production (28.66 mg/g of dry biomass) was observed at temperature 30°C, pH 5.0 and 1 g/l of initial cellular concentration after 2 h of fermentation. Agitation and aeration rate did not influence on RNA concentration (p >0.05). Maximum lactose consumption (98.7%) and biomass production (6.0 g/l) was observed after 12 h of incubation. This study proves that cheese whey can be used as an adequate medium for RNA production by K. marxianus under the optimized conditions at industrial scale.

Effects of synthesis conditions on the nanostructre of CexZr1-xO2 mesoporous ceramics.

CexZr1-xO2 (0.5 ≤ x ≤ 0.9) were synthesized with Zr and Ce chloride precursors, using the triblock copolymer Pluronic P123 and HCl (2 mol/L). The pH adjustment was performed in two ways: synthesis A used 11.4 mL of a NH4OH solution added at once to the initial mixture, composed by metal precursors and template in HCl; synthesis B was done by dripping slowly until the change of pH value (between 3 and 6). In this work, CexZr1-xO2 samples synthesized by these two processes are compared. The effects of pH values in materials characteristics were also evaluated. These samples were analysed by X-Ray Diffraction (XRD) with Rietveld refinement, and Nitrogen Adsorption/Desorption. In both processes, the studied materials presented two crystalline phases of CexZr1-xO2 solid solution: cubic and tetragonal. The synthesis A also presented a tetragonal phase of ZrO2. The average crystallite size and the Brunauer- Emmett-Teller (BET) surface area are bigger in process A. Both processes give samples with a mesoporous structure.

The structure of planktonic communities under variable coastal upwelling conditions off Cape Ghir (31ºN), in the Canary Current System (NW Africa)

Universidad de Las Palmas de Gran Canaria. Facultad de Ciencias del Mar. Programa de doctorado en Oceanografía. Diploma de Estudios Avanzados

Bioremediation of PAHs - Limitations and soultions

Introduction 1.1 Occurrence of polycyclic aromatic hydrocarbons (PAH) in the environment Worldwide industrial and agricultural developments have released a large number of natural and synthetic hazardous compounds into the environment due to careless waste disposal, illegal waste dumping and accidental spills. As a result, there are numerous sites in the world that require cleanup of soils and groundwater. Polycyclic aromatic hydrocarbons (PAHs) are one of the major groups of these contaminants (Da Silva et al., 2003). PAHs constitute a diverse class of organic compounds consisting of two or more aromatic rings with various structural configurations (Prabhu and Phale, 2003). Being a derivative of benzene, PAHs are thermodynamically stable. In addition, these chemicals tend to adhere to particle surfaces, such as soils, because of their low water solubility and strong hydrophobicity, and this results in greater persistence under natural conditions. This persistence coupled with their potential carcinogenicity makes PAHs problematic environmental contaminants (Cerniglia, 1992; Sutherland, 1992). PAHs are widely found in high concentrations at many industrial sites, particularly those associated with petroleum, gas production and wood preserving industries (Wilson and Jones, 1993). 1.2 Remediation technologies Conventional techniques used for the remediation of soil polluted with organic contaminants include excavation of the contaminated soil and disposal to a landfill or capping - containment - of the contaminated areas of a site. These methods have some drawbacks. The first method simply moves the contamination elsewhere and may create significant risks in the excavation, handling and transport of hazardous material. Additionally, it is very difficult and increasingly expensive to find new landfill sites for the final disposal of the material. The cap and containment method is only an interim solution since the contamination remains on site, requiring monitoring and maintenance of the isolation barriers long into the future, with all the associated costs and potential liability. A better approach than these traditional methods is to completely destroy the pollutants, if possible, or transform them into harmless substances. Some technologies that have been used are high-temperature incineration and various types of chemical decomposition (for example, base-catalyzed dechlorination, UV oxidation). However, these methods have significant disadvantages, principally their technological complexity, high cost , and the lack of public acceptance. Bioremediation, on the contrast, is a promising option for the complete removal and destruction of contaminants. 1.3 Bioremediation of PAH contaminated soil & groundwater Bioremediation is the use of living organisms, primarily microorganisms, to degrade or detoxify hazardous wastes into harmless substances such as carbon dioxide, water and cell biomass Most PAHs are biodegradable unter natural conditions (Da Silva et al., 2003; Meysami and Baheri, 2003) and bioremediation for cleanup of PAH wastes has been extensively studied at both laboratory and commercial levels- It has been implemented at a number of contaminated sites, including the cleanup of the Exxon Valdez oil spill in Prince William Sound, Alaska in 1989, the Mega Borg spill off the Texas coast in 1990 and the Burgan Oil Field, Kuwait in 1994 (Purwaningsih, 2002). Different strategies for PAH bioremediation, such as in situ , ex situ or on site bioremediation were developed in recent years. In situ bioremediation is a technique that is applied to soil and groundwater at the site without removing the contaminated soil or groundwater, based on the provision of optimum conditions for microbiological contaminant breakdown.. Ex situ bioremediation of PAHs, on the other hand, is a technique applied to soil and groundwater which has been removed from the site via excavation (soil) or pumping (water). Hazardous contaminants are converted in controlled bioreactors into harmless compounds in an efficient manner. 1.4 Bioavailability of PAH in the subsurface Frequently, PAH contamination in the environment is occurs as contaminants that are sorbed onto soilparticles rather than in phase (NAPL, non aqueous phase liquids). It is known that the biodegradation rate of most PAHs sorbed onto soil is far lower than rates measured in solution cultures of microorganisms with pure solid pollutants (Alexander and Scow, 1989; Hamaker, 1972). It is generally believed that only that fraction of PAHs dissolved in the solution can be metabolized by microorganisms in soil. The amount of contaminant that can be readily taken up and degraded by microorganisms is defined as bioavailability (Bosma et al., 1997; Maier, 2000). Two phenomena have been suggested to cause the low bioavailability of PAHs in soil (Danielsson, 2000). The first one is strong adsorption of the contaminants to the soil constituents which then leads to very slow release rates of contaminants to the aqueous phase. Sorption is often well correlated with soil organic matter content (Means, 1980) and significantly reduces biodegradation (Manilal and Alexander, 1991). The second phenomenon is slow mass transfer of pollutants, such as pore diffusion in the soil aggregates or diffusion in the organic matter in the soil. The complex set of these physical, chemical and biological processes is schematically illustrated in Figure 1. As shown in Figure 1, biodegradation processes are taking place in the soil solution while diffusion processes occur in the narrow pores in and between soil aggregates (Danielsson, 2000). Seemingly contradictory studies can be found in the literature that indicate the rate and final extent of metabolism may be either lower or higher for sorbed PAHs by soil than those for pure PAHs (Van Loosdrecht et al., 1990). These contrasting results demonstrate that the bioavailability of organic contaminants sorbed onto soil is far from being well understood. Besides bioavailability, there are several other factors influencing the rate and extent of biodegradation of PAHs in soil including microbial population characteristics, physical and chemical properties of PAHs and environmental factors (temperature, moisture, pH, degree of contamination). Figure 1: Schematic diagram showing possible rate-limiting processes during bioremediation of hydrophobic organic contaminants in a contaminated soil-water system (not to scale) (Danielsson, 2000). 1.5 Increasing the bioavailability of PAH in soil Attempts to improve the biodegradation of PAHs in soil by increasing their bioavailability include the use of surfactants , solvents or solubility enhancers.. However, introduction of synthetic surfactant may result in the addition of one more pollutant. (Wang and Brusseau, 1993).A study conducted by Mulder et al. showed that the introduction of hydropropyl-ß-cyclodextrin (HPCD), a well-known PAH solubility enhancer, significantly increased the solubilization of PAHs although it did not improve the biodegradation rate of PAHs (Mulder et al., 1998), indicating that further research is required in order to develop a feasible and efficient remediation method. Enhancing the extent of PAHs mass transfer from the soil phase to the liquid might prove an efficient and environmentally low-risk alternative way of addressing the problem of slow PAH biodegradation in soil.