The possible role of microcystin (D-Leu1 MC-LR) as an antioxidant on Microcystis aeruginosa (Cyanophyceae). In vitro and in vivo evidence.

Microcystins constitute a serious threat to the quality of drinking water worldwide. However, the eco-physiological role of them is not completely known and it is suggested that toxins can play a role in the antioxidant protection. The objective of this study was to evaluate the microcystin antioxidant capacity in vitro by Electronic Paramagnetic Resonance, highly specific for the different reactive oxygen species and in vivo by 7 days exposure of Microcystis aeruginosa to high (29 °C) temperature in addition to a 26 °C control condition. An effective in vitro antioxidant activity was observed for [D-Leu1]MC-LR against hydrosoluble radicals. As far as we know, this is the first in vitro record of the role of MC as antioxidant. In addition, a significant increase in cellular biomass was observed under 26 °C in cultures with [D-Leu1]MC-LR supplementation in coincidence with a significant decrease of reactive species. For cultures at 29 °C, the antioxidant role of toxins was inconclusive probably due to the presence of different reactive species generated during the experiment. Thus, MC could scavenge certain reactive species associated with the antioxidant role of CAT or the OH content by SOD activity (not measured) and then CAT activity could be lower in the presence of MC. Reinforcing our hypothesis, the [D-Leu1]MC-LR consumption after 7 days was significantly higher in cells with [D-Leu1]MC-LR supplementation in both 26 °C and 29 °C.When the production of reactive species was controlled by the scavenger activity of antioxidants plus MC, cells avoided the potential oxidative damage and started with exponential growth.

Assembly of chitosan support matrix with gallic acid-functionalized nanoparticles.

The incorporation of gallic acid (GA) in a direct way or into nanoparticles included in chitosan edible films appears as a suitable approach to increase its preservation upon adverse conditions. The addition of nanoparticles to chitosan-based matrices resulted in improvements in their solubility, swelling, and mechanical properties. It is worth noting that by means of the nanoencapsulation, the release process can be modulated in relation to the delivery of GA included directly in the matrix, releasing the agent at a lower rate for a longer time. Films containing functionalized nanoparticles are promising as a means to develop tailor-made support matrices for improving the shelf stability of the included active compound. From the point of view of the antimicrobial activity, all studied films showed bacteriostatic activity against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Salmonella spp. and Candida vinaria. The results suggest that active films with nanoparticles could be a potential candidate for the support and controlled release of active compounds such as gallic acid.

Changes induced by UV radiation in the presence of sodium benzoate in films formulated with polyvinyl alcohol and carboxymethyl cellulose.

This work was focused on: i) developing single and blend films based on carboxymethyl cellulose (CMC) and polyvinyl alcohol (PVOH) studying their properties, ii) analyzing the interactions between CMC and PVOH and their modifications UV-induced in the presence of sodium benzoate (SB), and iii) evaluating the antimicrobial capacity of blend films containing SB with and without UV treatment. Once the blend films with SB were exposed to UV radiation, they exhibited lower moisture content as well as a greater elongation at break and rougher surfaces compared to those without treatment. Considering oxygen barrier properties, the low values obtained would allow their application as packaging with selective oxygen permeability. Moreover, the characteristics of the amorphous phase of the matrix prevailed with a rearrangement of the structure of the polymer chain, causing a decrease of the crystallinity degree. These results were supported by X-rays and DSC analysis. FT-IR spectra reflected some degree of polymer-polymer interaction at a molecular level in the amorphous regions. The incorporation of sodium benzoate combined with UV treatment in blend films was positive from the microbial point of view because of the growth inhibition of a wide spectrum of microorganisms. From a physicochemical perspective, the UV treatment of films also changed their morphology rendering them more insoluble in water, turning the functionalized blend films into a potential material to be applied as food packaging.

Comparison of channeling contrast between ion and electron images.

Ion channeling contrast (iCC) and electron channeling contrast (eCC) are caused by variation in signal resulting from changes in the angle of the incident beam and the crystal lattice with respect to the target. iCC is directly influenced by the incident ion range in crystalline materials. The ion range is larger for low-index crystal orientated grains, resulting in the emission of fewer secondary electrons at the surface yielding a lower signal. Ions are stopped closer to the surface for off-axis grains, resulting in the emission of many secondary electrons yielding a higher signal. Conversely, backscattered electrons (BSEs) are the primary contribution to eCC. BSEs are diffracted or channeled to form an electron channeling pattern (ECP). The BSE emission of the ECP peaks when the electron beam is normal to the surface of an on-axis grain, and therefore a bright signal is observed. Thus, iCC and eCC images yield inverse contrast behavior for on-axis oriented grains. Since there is a critical angle associated with particle channeling, accurately determining grain boundary locations require the acquisition of multiple images obtained at different tilt conditions.

Occurrence of toxic cyanobacterial blooms in rio de la plata estuary, Argentina: field study and data analysis.

Water samples were collected during 3 years (2004-2007) at three sampling sites in the Rio de la Plata estuary. Thirteen biological, physical, and chemical parameters were determined on the water samples. The presence of microcystin-LR in the reservoir samples, and also in domestic water samples, was confirmed and quantified. Microcystin-LR concentration ranged between 0.02 and 8.6 µg.L(-1). Principal components analysis was used to identify the factors promoting cyanobacteria growth. The proliferation of cyanobacteria was accompanied by the presence of high total and fecal coliforms bacteria (>1500 MNP/100 mL), temperature ≥25°C, and total phosphorus content ≥1.24 mg·L(-1). The observed fluctuating patterns of Microcystis aeruginosa, total coliforms, and Microcystin-LR were also described by probabilistic models based on the log-normal and extreme value distributions. The sampling sites were compared in terms of the distribution parameters and the probability of observing high concentrations for Microcystis aeruginosa, total coliforms, and microcystin-LR concentration.

Non-monotonic material contrast in scanning ion and scanning electron images.

30keV Ga(+) focused ion beam induced secondary electron (iSE) imaging was used to determine the relative contrast between several materials. The iSE signal compared from C, Si, Al, Ti, Cr, Ni, Cu, Mo, Ag, and W metal layers does not decrease with an increase in target atomic number Z(2), and shows a non-monotonic relationship between contrast and Z(2). The non-monotonic relationship is attributed to periodic fluctuations of the stopping power and sputter yield inherent to the ion-solid interactions. In addition, material contrast from electron-induced secondary electron (eSE) and backscattered electron (BSE) images using scanning electron microscopy (SEM) also shows non-monotonic contrast as a function of Z(2), following the periodic behavior of the stopping power for electron-solid interactions. A comparison of the iSE and eSE results shows similar relative contrast between the metal layers, and not complementary contrast as conventionally understood. These similarities in the contrast behavior can be attributed to similarities in the periodic and non-monotonic function defined by incident particle-solid interaction theory.

High-temperature interface superconductivity between metallic and insulating copper oxides.

The realization of high-transition-temperature (high-T(c)) superconductivity confined to nanometre-sized interfaces has been a long-standing goal because of potential applications and the opportunity to study quantum phenomena in reduced dimensions. This has been, however, a challenging target: in conventional metals, the high electron density restricts interface effects (such as carrier depletion or accumulation) to a region much narrower than the coherence length, which is the scale necessary for superconductivity to occur. By contrast, in copper oxides the carrier density is low whereas T(c) is high and the coherence length very short, which provides an opportunity-but at a price: the interface must be atomically perfect. Here we report superconductivity in bilayers consisting of an insulator (La(2)CuO(4)) and a metal (La(1.55)Sr(0.45)CuO(4)), neither of which is superconducting in isolation. In these bilayers, T(c) is either approximately 15 K or approximately 30 K, depending on the layering sequence. This highly robust phenomenon is confined within 2-3 nm of the interface. If such a bilayer is exposed to ozone, T(c) exceeds 50 K, and this enhanced superconductivity is also shown to originate from an interface layer about 1-2 unit cells thick. Enhancement of T(c) in bilayer systems was observed previously but the essential role of the interface was not recognized at the time.

Mathematical modelling of microbial growth in ground beef from Argentina. Effect of lactic acid addition, temperature and packaging film.

The effects of: (i) storage temperature (0, 4 and 10°C), (ii) gaseous permeability of the packaging film (polyethylene and EVA SARAN EVA for vacuum packaging), and (iii) natural beef pH (5.6, 5.8 and 6.1) on the growth of different bacteria isolated from beef muscle were examined. The bacteria were Klebsiella, Pseudomonas sp. and Escherichia coli. Microbial growth was modelled using Gompertz and linear equations. The effects of temperature on microbial growth rate (µ) and on lag phase duration were modelled using an Arrhenius type equation. In polyethylene, E. coli was the microorganism, that showed the highest µ values and also the greatest effect of pH on µ, especially in samples stored at 4 and 10°C. In the case of Klebsiella sp., neither pH nor temperature had marked effects on µ and on LPD. In ESE film, µ of all the microorganisms were less affected by pH and temperature than in polyethylene. In ESE film E. coli showed the highest effect of pH on µ, at 4 and 10°C. LPD increased significantly with respect to the values in polyethylene, with Klebsiella sp., showing the highest values of LPD, followed by E. coli. Experiments in ground beef with added lactic acid producing a decrease of the original muscle pH from 6.1 to 5.6 showed that the kinetic parameters of the microbial flora did not differ significantly from those of beef samples in which the original pH was 5.6.

Inhibitory effect of a surfactant on pure cultures of a filamentous and a floc forming micro-organism.

Activated sludge is the most widely used biological process for wastewater treatment. Inorganic and organic compounds are removed by a biotic community in the aeration basin. Problems of these systems are loss of settleability and poor sludge compaction due to excessive growth of filamentous micro-organisms. The filamentous bulking can be controlled by the addition of chemical agents. Strong oxidants, such as chlorine, are utilized to eliminate filamentous bacteria; however, these substances also tend to attack floc-forming bacteria and to cause process breakdown. Besides, chlorine may become hazardous owing to the formation of chemical products as chloramines. Surfactant addition constitutes an interesting alternative for the control of filamentous bulking. In this work the effect of a surfactant Triton X-100 (octylphenol ethoxylate), on the respiratory activity (RA) of pure cultures of a filamentous (Sphaerotilus natans) and a floc-former microorganism (Acinetobacter anitratus) was evaluated. In the concentration range tested (60-220 mg l(-1)), the surfactant was observed to exhibit high RA specific inhibition of the filamentous micro-organism with no significant effect on the floc-forming bacteria. Light microscopy observations showed that the surfactant induced cell lysis, leaving only empty sheaths in the case of filamentous micro-organisms. A kinetic equation to predict the microbial RA fraction of a S. natans pure culture as a function of surfactant concentration and contact time was proposed. The effect of Triton X-100 on the inactivation of pure cultures of both micro-organisms was compared to that of chlorine. Triton X-100 results were adequate to eliminate filamentous bacteria emerging as an alternative for filamentous bulking treatment.

Kinetics of ethanol degradation in forensic blood samples.

To determine ethanol in human post-mortem blood samples is problematic, largely due to the inappropriate and variable methods of preserving and storing, which can cause decomposition and loss of alcohol concentration. In this study, four crucial parameters of sample conservation were studied: temperature (T), percentage of air chamber in container (%CA), ethanol concentration in blood and post-mortem time. Blood samples from post-mortem cases were stored under different conditions (ethanol levels were known in all cases); factorial design variables: (%CA) 0, 5, 20, 35, 65%; storage temperature: 25, 4 and -10 degrees C; in a total of 15 experiments. No preserving agent was used in samples. Quantification of ethanol in blood was carried out by gas chromatography with head-space FID detector. Initial ethanol concentration ranged from 0.50 to 4.30 g/L. The kinetics of degradation observed was pseudo-first-order. The parameter that characterised the kinetics of ethanol degradation (k(0)) ranged from (4 x 10(-4) and 5.0 x 10(-1) day(-1)), depending on storage conditions. A strong dependence between ethanol degradation and the content of the air chamber was observed and this dependence was found to be stronger than that between degradation and temperature; there was an experimental relation between (k(0)) and (%CA). Activation energy for different conditions, i.e. 0, 5, 20, 35 and 65 (%CA), were calculated and contour plots were made. A mathematical equation relating air chamber, temperature and ethanol concentration at a certain time was determined. This equation allowed estimation of initial concentrations of ethanol with minimal error. A good correlation between experimental data and data calculated with the equation was obtained (r(2) = 0.9998). The best storage conditions were: 0% CA and storage at -10 degrees C, obtaining an ethanol degradation of 0.01% after 15 days. However, 33% of ethanol degradation was obtained with 35% CA at 25 degrees C after 15 days. This equation is useful in forensic cases in which original concentration of ethanol has to be estimated under different sample storage conditions.

The correlation between ion beam/material interactions and practical FIB specimen preparation.

The focused ion beam (FIB) tool has been successfully used as both a stand alone analytical instrument and a means to prepare specimens for subsequent analysis by SEM, TEM, SIMS, XPS, and AUGER. In this work, special emphasis is given to TEM specimen preparation by the FIB lift-out technique. The fundamental ion/solid interactions that govern the FIB milling process are examined and discussed with respect to the preparation of electron transparent membranes. TRIM, a Monte Carlo simulation code, is used to physically model variables that influence FIB sputtering behavior. The results of such computer generated models are compared with empirical observations in a number of materials processed with an FEI 611 FIB workstation. The roles of incident ion attack angle, beam current, trench geometry, raster pattern, and target-material-dependent removal rates are considered. These interrelationships are used to explain observed phenomena and predict expected milling behaviors, thus increasing the potential for the FIB to be used more efficiently with reproducible results.

Competitive growth kinetics of Sphaerotilus natans and Acinetobacter anitratus.

Growth kinetics of Sphaerotilus natans and Acinetobacter anitratus (strain E932) in pure and mixed cultures were analysed. In order to determine mixed cultures biomass composition, a quantitative image analysis technique was developed. Pure culture studies showed that for dilution rates less than 0.188 h(-1), the filamentous micro-organism will predominate leading to bulking phenomena. By using the developed technique to determine biomass composition, mixed culture experiments showed that changes in the dilution rate modify the microbial composition of the biomass determining which micro-organism predominate. The stated equations that predict the predominance of S. natans at low dilution rates agree satisfactorily with the obtained results.

Hydrogen cyanide and carbon monoxide in blood of convicted dead in a polyurethane combustion: a proposition for the data analysis.

Carbon monoxide is a well-known toxic component in fire atmospheres. However, the importance of hydrogen cyanide as a toxic agent in fire causalities is under discussion. A tragic polyurethane mattress fire provoked death of 35 convicts in a prison (Unit I, Olmos, Penitenciary Service of Buenos Aires Province, Argentina), in 1990. There is no report of any investigation carried out with such a large amount of victims in Argentina. Carboxihemoglobin (COHb) and hydrogen cyanide (HCN) were quantified in victims blood to elucidate the cause of the death. Saturation of COHb ranged between 4 and 18%, and HCN 2.0-7.2mg/l. These latter values were higher than the lethal levels reported in literature. Other toxic components routinely measured (ethanol, methanol, aldehydes and other volatile compounds) gave negative results on the 35 cases. Neither drugs of abuse nor psychotropics were detected. Statistical chi(2) analysis was applied to find differences between HCN and COHb concentrations. Saturation of COHb and HCN in blood were not independent variables (chi(2)=8.25). Moreover, the ratio COHb/HCN was constant (0.47+/-0.04). In order to evaluate the contribution of each toxic to the diagnosis, a lethal index was defined for each toxic (LI(CO) and LI(HCN)). The most probable cause of death could be inferred by a suitable plot of both indexes. The results indicated that death in the 35 fire victims was probably caused by HCN, generated during the extensive polyurethane decomposition provoked by a rapid increase of temperature.

Modeling the aerobic growth and decline of Staphylococcus aureus as affected by pH and potassium sorbate concentration.

The effects of pH (5.0, 5.2, 5.4, 5.6, and 5,8) and concentration of potassium sorbate (10.0 and 16.6 mM) at two water activity values (0.90 and 0.92) on the aerobic growth and decline of Staphylococcus aureus ATCC 6538P, 196-E, and FDA-C243 were studied using brain-heart infusion broth. The inoculum was approximately 4 to 5 log CFU/ml, and the incubation temperature was 30 degrees C. Samples were periodically enumerated on tryptic soy agar. The Gompertz model was used to obtain microbial growth parameters, specific growth rate was obtained as a derived parameter, and the inhibition index was calculated. A linear model was fitted in cases of bacteriostatic or bactericidal action of the treatment. The ATCC 6538P strain showed the highest resistance in the range of tested conditions. Microbial behavior was modeled considering the main controlling factors, and a response surface methodology was used to determine the effects of undissociated acid concentration and pH. These results can be used to establish treatment conditions for microorganism growth or inhibition.

Applications of the FIB lift-out technique for TEM specimen preparation.

A site-specific technique for cross-section transmission electron microscopy specimen preparation of difficult materials is presented. A focused ion beam was used to slice an electron transparent membrane from a specific area of interest within a bulk sample. Micromanipulation lift-out procedures were then used to transport the electron-transparent specimen to a carbon-coated copper grid for subsequent TEM analysis. The FIB (focused ion beam) lift-out technique is a fast method for the preparation of site-specific TEM specimens. The versatility of this technique is demonstrated by presenting cross-sectioned TEM specimens from several types of materials systems, including a multi-layered integrated circuit on a Si substrate, a galvanized steel, a polycrystalline SiC ceramic fiber, and a ZnSe optical ceramic. These specimens have both complex surface geometry and interfaces with complex chemistry. FIB milling was performed sequentially through different layers of cross-sectioned materials so that preferential sputtering was not a factor in preparing TEM specimens. The FIB lift-out method for TEM analysis is a useful technique for the study of complex materials systems for TEM analysis.

Mathematical modelling of microbial growth in packaged refrigerated beef stored at different temperatures.

Gompertz and logistic models were fitted to experimental counts of microorganisms growing in beef stored at 0, 4, 7, 9 and 10 degrees C. Samples were packaged in polyethylene (high gaseous permeability) and in EVA/SARAN/EVA (low gaseous permeability) films, being EVA ethyl vinyl acetate and SARAN polyvinyl and polyvinylidene chloride copolymer. Lag phase duration (LPD) and specific growth rate (mu) were obtained as derived parameters for lactic acid bacteria, Enterobacteriaceae, Pseudomonas sp. and psychrotrophic microorganisms. The reciprocal of LPD was fitted to an Arrhenius type equation; LPD of lactic acid bacteria showed a marked dependence on temperature, with activation energy values (ELPD) of 222.2 and 216.9 kJ/mol for polyethylene and ESE respectively. The effect of initial microbial population at different storage temperatures on adaptation period was analyzed. As the initial microbial population increased, adaptation period decreased for all studied microorganisms and for both packaging films. The effect of temperature on specific growth rate was better interpreted by the Arrhenius model than by the linear or the square root equations. Psychrotrophic microorganisms in beef showed the highest activation energy values for specific growth rate (E mu) in both packaging films, being E mu 85.50 and 103.10 KJ/mol for polyethylene and ESE film respectively. In both films, Enterobacteriaceae showed the lowest E mu values, being 15.33 and 59.89 kJ/mol in ESE and polyethylene respectively. The final number of microorganisms (maximum population density) did not show significant changes with storage temperature.

Chemical Preservatives Action on Microbial Growth in a Model System of Refrigerated Prepeeled Potatoes.

The effect of different concentrations of citric acid and ascorbic acid (applied individually or in mixture's) on microbial growth in potato homogenate was analyzed and compared to the sodium bisulfite action during storage at 4°C in low gaseous permeability films. These experiments allowed one to simulate the behavior of prepeeled potatoes but with a known amount of added preservative to evaluate additive or synergic effects. Total viable microorganisms, Enterobacteriaceae , Pseudomonas sp., Lactobacillus sp., molds, yeasts, Clostridium sulfite reducers, psychrotropic microorganisms, and aerobic and anaerobic viable spores were analyzed during storage time. Inhibition indexes produced by the tested preservatives were calculated for the different microorganisms. Sodium bisulfite solutions (100 ppm) had no inhibitory effect. Concentrations of 3,500 ppm citric acid and 10,000 ppm ascorbic acid showed antimicrobial action as well as mixtures of citric acid/ascorbic acid of the following compositions (in terms of total acids concentration): 2,700/2,000, 3,500/2,000, and 2,700/3,000 ppm. A higher effect on Enterobacteriaceae was observed in comparison with other microorganisms. The apparent synergic effect of these acids when they were applied together was demonstrated to be actually an additive effect when concentrations of undissociated acid in the mixtures were considered instead of total concentration.

[Growth of Staphylococcus aureus in solid media with a water activity less than 0.86]. / Crecimiento de Staphylococcus aureus en medios sólidos de actividad acuosa inferior a 0.86.

The purpose of this study was to determine whether S. aureus could grow in solid substrates with aw 0.843 adjusted by water adsorption. The media used included dehydrated powder of pork, beef and milk as basic substrates in order to compensate an eventual deficiency; additional source of nutrients was added. The tested strains were ATCC 6538 P, FDA-C243, NA1 and FM1. It was observed that no strain grew at 30 degrees C in those media at aw 0.843. These data were compared with others which indicate that S. aureus could grow al aw 0.83-0.84. The possible reasons for the difference in the minimal aw value reported to be required for the growth of S. aureus are discussed.

Crecimiento de Staphylococcus aureus en medios sólidos de actividad acuosa inferior a 0.86. / [Growth of Staphylococcus aureus in solid media with a water activity less than 0.86]

The purpose of this study was to determine whether S. aureus could grow in solid substrates with aw 0.843 adjusted by water adsorption. The media used included dehydrated powder of pork, beef and milk as basic substrates in order to compensate an eventual deficiency; additional source of nutrients was added. The tested strains were ATCC 6538 P, FDA-C243, NA1 and FM1. It was observed that no strain grew at 30 degrees C in those media at aw 0.843. These data were compared with others which indicate that S. aureus could grow al aw 0.83-0.84. The possible reasons for the difference in the minimal aw value reported to be required for the growth of S. aureus are discussed.