Photodynamic inactivation of planktonic cultures and biofilms of Candida albicans mediated by aluminum-chloride-phthalocyanine entrapped in nanoemulsions

New drug delivery systems, such as nanoemulsions (NE), have been developed to allow the use of hydrophobic drugs on the antimicrobial photodynamic therapy. This study evaluated the photodynamic potential of aluminum-chloride- phthalocyanine (ClAlPc) entrapped in cationic and anionic NE to inactivate Candida albicans planktonic cultures and biofilm compared with free ClAlPc. Fungal suspensions were treated with different delivery systems containing ClAlPc and light emitting diode. For planktonic suspensions, colonies were counted and cell metabolism was evaluated by XTT assay. Flow cytometry evaluated cell membrane damage. For biofilms, the metabolic activity was evaluated by XTT and ClAlPc distribution through biofilms was analyzed by confocal laser scanning microscopy (CLSM). Fungal viability was dependent on the delivery system, superficial charge and light dose. Free ClAlPc caused photokilling of the yeast when combined with 100 J cm-2. Cationic NE-ClAlPc reduced significantly both colony counts and cell metabolism (P < 0.05). In addition, cationic NE-ClAlPc and free ClAlPc caused significant damage to the cell membrane (P < 0.05). For the biofilms, cationic NE-ClAlPc reduced cell metabolism by 70%. Anionic NE-ClAlPc did not present antifungal activity. CLSM showed different accumulation on biofilms between the delivery systems. Although NE system showed a lower activity for planktonic culture, cationic NE-ClAlPc showed better results for Candida biofilms. Candida albicans biofilm overview after 30 min of contact with free ClAlPc. This study presents the photodynamic potential of aluminum-chloride-phthalocyanine (ClAlPc) entrapped in cationic and anionic nanoemulsions (NE) to inactivate C. albicans planktonic cultures and biofilm comparing with free ClAlPc. The photodynamic effect was dependent on the delivery system, superficial charge and light dose. Cationic NE-ClAlPc and free ClAlPc caused significant reduction in colony counts, cell metabolism and damage to the cell membrane (P < 0.05). However, only the free ClAlPc was able to cause photokilling of the yeast. The anionic NE-ClAlPc did not present antifungal activity. Although NE system showed a lower activity for planktonic culture, cationic NE-ClAlPc showed better results for Candida biofilms. © 2012 Wiley Periodicals, Inc. Photochemistry and Photobiology © 2012 The American Society of Photobiology.

Evaluation of the bone healing process utilizing platelet-rich plasma activated by thrombin and calcium chloride: A histologic study in rabbit calvaria

To evaluate the bone healing of defects filled with particulate bone graft in combination with platelet-rich plasma (PRP), added with a mixture of calcium chloride and thrombin or just calcium chloride. Two 5-mm bone defects were created in the calvaria of 24 rabbits. Each defect was filled with particulate bone graft and PRP. In one defect the PRP was activated by a mixture of calcium chloride and thrombin; in the other, PRP was activated by calcium chloride only. The animals were euthanized 1, 2, 4, and 8 weeks after the surgeries, and the calvaria was submitted to histologic processing for histomorphometric analysis. The qualitative analysis has shown that both defects presented the same histologic characteristics so that a better organized, more mature, and well-vascularized bone tissue was noticed in the eighth week. A good bone repair was achieved using either the mixture of calcium chloride and thrombin or the calcium chloride alone as a restarting agent of the coagulation process.

Biomimetic sensor based on 5,10,15,20-tetrakis(pentafluorophenyl)-21H,23H- porphyrin iron (III) chloride and MWCNT for selective detection of 2,4-D

Exploitation of the electronic properties of carbon nanotubes for the development of voltammetric and amperometric sensors to monitor analytes of environmental relevance has increased in recent years. This work reports the development of a biomimetic sensor based on a carbon paste modified with 5,10,15,20-tetrakis(pentafluorophenyl)-21H,23H-porphyrin iron (III) chloride (a biomimetic catalyst of the P450 enzyme) and multi-wall carbon nanotubes (MWCNT), for the sensitive and selective detection of the herbicide 2,4- dichlorophenoxyacetic acid (2,4-D). The sensor was evaluated using cyclic voltammetry and amperometry, for electrochemical characterization and quantification purposes, respectively. Amperometric analyses were carried out at -100 mV vs. Ag/AgCl(KClsat), using a 0.1 mol L-1 phosphate buffer solution at pH 6.0 as the support electrolyte. Under these optimized analytical conditions, the sensor showed a linear response between 9.9 × 10-6 and 1.4 × 10-4 mol L-1, a sensitivity of 1.8 × 104 (±429) μA L mol -1, and limits of detection and quantification of 2.1 × 10 -6 and 6.8 × 10-6 mol L-1, respectively. The incorporation of functionalized MWCNT in the carbon paste resulted in a 10-fold increase in the response, compared to that of the biomimetic sensor without MWCNT. In addition, the low applied potential (-100 mV) used to obtain high sensitivity also contributed to the excellent selectivity of the proposed sensor. The viability of the application of this sensor for analysis of soil samples was confirmed by satisfactory recovery values, with a mean of 96% and RSD of 2.1% (n = 3). © 2013 Elsevier B.V.

Baclofen into the lateral parabrachial nucleus induces hypertonic sodium chloride intake during cell dehydration

Background: Activation of GABAB receptors with baclofen into the lateral parabrachial nucleus (LPBN) induces ingestion of water and 0.3 M NaCl in fluid replete rats. However, up to now, no study has investigated the effects of baclofen injected alone or combined with GABAB receptor antagonist into the LPBN on water and 0.3 M NaCl intake in rats with increased plasma osmolarity (rats treated with an intragastric load of 2 M NaCl). Male Wistar rats with stainless steel cannulas implanted bilaterally into the LPBN were used.Results: In fluid replete rats, baclofen (0.5 nmol/0.2 μl), bilaterally injected into the LPBN, induced ingestion of 0.3 M NaCl (14.3 ± 4.1 vs. saline: 0.2 ± 0.2 ml/210 min) and water (7.1 ± 2.9 vs. saline: 0.6 ± 0.5 ml/210 min). In cell-dehydrated rats, bilateral injections of baclofen (0.5 and 1.0 nmol/0.2 μl) into the LPBN induced an increase of 0.3 M NaCl intake (15.6 ± 5.7 and 21.5 ± 3.5 ml/210 min, respectively, vs. saline: 1.7 ± 0.8 ml/210 min) and an early inhibition of water intake (3.5 ± 1.4 and 6.7 ± 2.1 ml/150 min, respectively, vs. saline: 9.2 ± 1.4 ml/150 min). The pretreatment of the LPBN with 2-hydroxysaclofen (GABAB antagonist, 5 nmol/0.2 μl) potentiated the effect of baclofen on 0.3 M NaCl intake in the first 90 min of test and did not modify the inhibition of water intake induced by baclofen in cell-dehydrated rats. Baclofen injected into the LPBN did not affect blood pressure and heart rate.Conclusions: Thus, injection of baclofen into the LPBN in cell-dehydrated rats induced ingestion of 0.3 M NaCl and inhibition of water intake, suggesting that even in a hyperosmotic situation, the blockade of LPBN inhibitory mechanisms with baclofen is enough to drive rats to drink hypertonic NaCl, an effect independent of changes in blood pressure. © 2013 Kimura et al.; licensee BioMed Central Ltd.

Influence of ethanol, acidity and chloride concentration on the corrosion resistance of AISI 316L stainless steel

The influence of ethanol, sulfuric acid and chloride on the corrosion resistance of 316L stainless steel was investigated by means of polarization curves and electrochemical impedance spectroscopy measurements. Over the studied range, the steel corrosion potential was independent of H2SO 4 and NaCl concentrations in aqueous solution. On the other hand, in solution containing 65 wt.% ethanol and 35 wt.% water, the corrosion potentials were higher than those obtained in aqueous solution. Besides, the steel corrosion potential was affected by the addition of H2SO4 and NaCl in solution. In solutions with and without ethanol, plus 0.35 wt.% NaCl, the presence of 1 wt.% H2SO4 inhibited the appearance of pitting corrosion. © 2013 Sociedade Brasileira de Química.

Photoelectrocatalysis based on Ti/TiO2 nanotubes removes toxic properties of the azo dyes Disperse Red 1, Disperse Red 13 and Disperse Orange 1 from aqueous chloride samples

This work describes the efficiency of photoelectrocatalysis based on Ti/TiO2 nanotubes in the degradation of the azo dyes Disperse Red 1, Disperse Red 13 and Disperse Orange 1 and to remove their toxic properties, as an alternative method for the treatment of effluents and water. For this purpose, the discoloration rate, total organic carbon (TOC) removal, and genotoxic, cytotoxic and mutagenic responses were determined, using the comet, micronucleus and cytotoxicity assays in HepG2 cells and the Salmonella mutagenicity assay. In a previous study it was found that the surfactant Emulsogen could contribute to the low mineralization of the dyes (60% after 4h of treatment), which, in turn, seems to account for the mutagenicity of the products generated. Thus this surfactant was not added to the chloride medium in order to avoid this interference. The photoelectrocatalytic method presented rapid discoloration and the TOC reduction was ≥87% after 240min of treatment, showing that photoelectrocatalysis is able to mineralize the dyes tested. The method was also efficient in removing the mutagenic activity and cytotoxic effects of these three dyes. Thus it was concluded that photoelectrocatalysis was a promising method for the treatment of aqueous samples. © 2013 Elsevier Ltd.

Effect of Na-chloride on the bioleaching of a chalcopyrite concentrate in shake flasks and stirred tank bioreactors

Oxidative dissolution of chalcopyrite at ambient temperatures is generally slow and subject to passivation, posing a major challenge for developing bioleaching applications for this recalcitrant mineral. Chloride is known to enhance the chemical leaching of chalcopyrite, but much of this effect has been demonstrated at elevated temperatures. This study was undertaken to test whether 100-200 mM Na-chloride enhances the chemical and bacterial leaching of chalcopyrite in shake flasks and stirred tank bioreactor conditions at mesophilic temperatures. Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans and abiotic controls were employed for the leaching experiments. Addition of Na-chloride to the bioleaching suspension inhibited the formation of secondary phases from chalcopyrite and decreased the Fe(III) precipitation. Neither elemental S nor secondary Cu-sulfides were detected in solid residues by X-ray diffraction. Chalcopyrite leaching was enhanced when the solution contained bacteria, ferrous iron and Na-chloride under low redox potential (< 450 mV) conditions. Scanning electron micrographs and energy-dispersive analysis of X-rays revealed the presence of precipitates that were identified as brushite and jarosites in solid residues. Minor amounts of gypsum may also have been present. Electrochemical analysis of solid residues was in concurrence of the differential effects between chemical controls, chloride ions, and bacteria. Electrochemical impedance spectroscopy was used to characterize interfacial changes on chalcopyrite surface caused by different bioleaching conditions. In abiotic controls, the impedance signal stabilized after 28 days, indicating the lack of changes on mineral surface thereafter, but with more resistive behavior than chalcopyrite itself. For bioleached samples, the signal suggested some capacitive response with time owing to the formation of less conductive precipitates. At Bode-phase angle plots (middle frequency), a new time constant was observed that was associated with the formation of jarosite, possibly also with minor amount or elemental S, although this intermediate could not be verified by XRD. Real impedance vs. frequency plots indicated that the bioleaching continued to modify the chalcopyrite/solution interface even after 42 days. © 2013 The Authors.

Cytotoxicity and genotoxicity of low doses of mercury chloride and methylmercury chloride on human lymphocytes in vitro

Mercury is a xenobiotic metal that is a highly deleterious environmental pollutant. The biotransformation of mercury chloride (HgCl2) into methylmercury chloride (CH3HgCl) in aquatic environments is well-known and humans are exposed by consumption of contaminated fish, shellfish and algae. The objective of the present study was to determine the changes induced in vitro by two mercury compounds (HgCl2 and CH3HgCl) in cultured human lymphocytes. Short-term human leukocyte cultures from 10 healthy donors (5 females and 5 males) were set-up by adding drops of whole blood in complete medium. Cultures were separately and simultaneously treated with low doses (0.1 to 1000 µg/l) of HgCl2 and CH3HgCl and incubated at 37ºC for 48 h. Genotoxicity was assessed by chromosome aberrations and polyploid cells. Mitotic index was used as a measure of cytotoxicity. A significant increase (P < 0.05) in the relative frequency of chromosome aberrations was observed for all concentrations of CH3HgCl when compared to control, whether alone or in an evident sinergistic combination with HgCl2. The frequency of polyploid cells was also significantly increased (P < 0.05) when compared to control after exposure to all concentrations of CH3HgCl alone or in combination with HgCl2. CH3HgCl significantly decreased (P < 0.05) the mitotic index at 100 and 1000 µg/l alone, and at 1, 10, 100, and 1000 µg/l when combined with HgCl2, showing a synergistic cytotoxic effect. Our data showed that low concentrations of CH3HgCl might be cytotoxic/genotoxic. Such effects may indicate early cellular changes with possible biological consequences and should be considered in the preliminary evaluation of the risks of populations exposed in vivo to low doses of mercury.

Losses of immunoreactive parvalbumin amacrine and immunoreactive alphaprotein kinase C bipolar cells caused by methylmercury chloride intoxication in the retina of the tropical fish Hoplias malabaricus

To quantify the effects of methylmercury (MeHg) on amacrine and on ON-bipolar cells in the retina, experiments were performed in MeHg-exposed groups of adult trahiras (Hoplias malabaricus) at two dose levels (2 and 6 µg/g, ip). The retinas of test and control groups were processed by mouse anti-parvalbumin and rabbit anti-_aprotein kinase C (_aPKC) immunocytochemistry. Morphology and soma location in the inner nuclear layer were used to identify immunoreactive parvalbumin (PV-IR) and _aPKC (_aPKC-IR) in wholemount preparations. Cell density, topography and isodensity maps were estimated using confocal images. PV-IR was detected in amacrine cells in the inner nuclear layer and in displaced amacrine cells from the ganglion cell layer, and _aPKC-IR was detected in ON-bipolar cells. The MeHg-treated group (6 µg/g) showed significant reduction of the ON-bipolar _aPKC-IR cell density (mean density = 1306 ± 393 cells/mm²) compared to control (1886 ± 892 cells/mm²; P < 0.001). The mean densities found for amacrine PV-IR cells in MeHg-treated retinas were 1040 ± 56 cells/mm² (2 µg/g) and 845 ± 82 cells/mm² (6 µg/g), also lower than control (1312 ± 31 cells/mm²; P < 0.05), differently from the data observed in displaced PV-IR amacrine cells. These results show that MeHg changed the PV-IR amacrine cell density in a dose-dependent way, and reduced the density of _aKC-IR bipolar cells at the dose of 6 µg/g. Further studies are needed to identify the physiological impact of these findings on visual function.

Assessing the effectiveness of 30% sodium chloride aqueous solution for the preservation of fixed anatomical specimens: a 5-year follow-up study

Anatomical specimens used in human or veterinary anatomy laboratories are usually prepared with formaldehyde (a cancerous and teratogenic substance), glycerin (an expensive and viscous fluid), or ethanol (which is flammable). This research aimed to verify the viability of an aqueous 30% sodium chloride solution for preservation of anatomical specimens previously fixed with formaldehyde. Anatomical specimens of ruminant, carnivorous, equine, swine and birds were used. All were previously fixed with an aqueous 20% formaldehyde solution and held for 7days in a 10% aqueous solution of the same active ingredient. During the first phase of the experiment, small specimens of animal tissue previously fixed in formaldehyde were distributed in vials with different concentrations of formaldehyde, with or without 30% sodium chloride solution, a group containing only 30% sodium chloride, and a control group containing only water. During this phase, no contamination was observed in any specimen containing 30% sodium chloride solution, whether alone or in combination with different concentrations of formaldehyde. In the second phase of the experiment, the 30% sodium chloride solution, found to be optimal in the first phase of the experiment, was tested for its long-term preservation properties. For a period of 5years, the preserved specimens were evaluated three times a week for visual contamination, odors, and changes in color and texture. There was no visual contamination or decay found in any specimen. Furthermore, no strange odors, or changes in color or softness were noted. The 30% sodium chloride solution was determined to be effective in the preservation of anatomic specimens previously fixed in formaldehyde.

Cetyltrimethylammonium chloride (CTAC) effect on the thermal stability of oxy-HbGp: Dynamic light scattering (DLS) and small angle X-ray scattering (SAXS) studies

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

The Effects of Weathering Exposure on the Physical, Mechanical, and Thermal Properties of High-density Polyethylene and Poly (Vinyl Chloride)

This paper presents results describing the physical, mechanical, and thermal properties (melt flow index - MFI and oxidative induction time - OIT) of high density polyethylene and poly (vinyl chloride) after weathering exposure (6, 12, 18, and 30 months). The materials exposed were geomembranes of two thicknesses: 1.0 and 2.0 mm (PVC) and 0.8 and 2.5 mm (HDPE). The climate parameters (average) obtained were 25 degrees C (temperature), 93 mm (precipitation), 66% (relative humidity), and 19 MJ/m(2). day (intensity of global radiation). Some results showed, for instance, that the behavior of the geomembranes changed after the exposures. A few minor variations in physical properties occurred. The density and thickness, for instance, varied 0.5-1.0% (average) for both the PVC and HDPE geomembranes. The mechanical properties changed as a function of the period of exposure. In general, some decreases were verified by the deformation of PVC. The samples became more rigid. In contrast, HDPE geomembranes became more ductile. Despite the variations in elasticity, some increases in deformability were verified. An MFI test showed some degradation in HDPE geomembranes. OIT tests revealed small values for both intact and exposed samples.

Nonlinear formulation based on FEM, Mazars damage criterion and Fick's law applied to failure assessment of reinforced concrete structures subjected to chloride ingress and reinforcements corrosion

Structural durability is an important design criterion, which must be assessed for every type of structure. In this regard, especial attention must be addressed to the durability of reinforced concrete (RC) structures. When RC structures are located in aggressive environments, its durability is strongly reduced by physical/chemical/mechanical processes that trigger the corrosion of reinforcements. Among these processes, the diffusion of chlorides is recognized as one of major responsible of corrosion phenomenon start. To accurate modelling the corrosion of reinforcements and to assess the durability of RC structures, a mechanical model that accounts realistically for both concrete and steel mechanical behaviour must be considered. In this context, this study presents a numerical nonlinear formulation based on the finite element method applied to structural analysis of RC structures subjected to chloride penetration and reinforcements corrosion. The physical nonlinearity of concrete is described by Mazars damage model whereas for reinforcements elastoplastic criteria are adopted. The steel loss along time due to corrosion is modelled using an empirical approach presented in literature and the chloride concentration growth along structural cover is represented by Fick's law. The proposed model is applied to analysis of bended structures. The results obtained by the proposed numerical approach are compared to responses available in literature in order to illustrate the evolution of structural resistant load after corrosion start. (C) 2014 Elsevier Ltd. All rights reserved.

Effect of calcium chloride on the pH values of several formulations of calcium hydroxide mixed with chlorhexidine digluconate

This study evaluated the effect of the addition of 5% calcium chlorite (CaCl2) on pH values in calcium hydroxide pastes (CH), with or without 2% chlorhexidine digluconate (CHX) used as vehicle, in several periods analysis. Polyethylene tubes were filled with CH mixed with water (G1), 2% CHX solution (G2) or gel (G3), or CHX solution or gel with 5% CaCl2 (G4 and G5, respectively). All tubes were individually immersed in distilled water. After 12, 24 hours, 7, 14 and 28 days, pH value was evaluated directly in water which the tubes were stored. Data were submitted to ANOVA and Tukey tests (α=0.05). In 24 hs and 14 days, pH values were similar to all groups. In 12 hs, the G1 presented lower pH value than other groups except to G4 (p < 0.05), and G4 presented lower pH value than G5 (p < 0.05). In 7 days, G1 presented lower pH value than G4 and G5 (p < 0.05). In 28 days, G1 and G5 presented lower pH values than G2 and G4 (p < 0.05) and among other groups there are no statistical differences (p > 0.05). The pH values increased in long-term analysis to all CH pastes. The association of 5% calcium chloride with 2% CHX solution as vehicle of CH paste provided a pH value increase in relation to CH mixed with distilled water. The CHX gel interfered negatively on pH value in comparison to CHX solution when mixed with CaCl2.

The association of chlorhexidine digluconate and calcium chloride to use as vehicle of a silicate calcium-based cement

Objective: The purpose of this study was to evaluate the influence of the addition of 2% chlorhexidine digluconate (CHX) associated with 5% calcium chloride (CaCl2 ) on antimicrobial activity, setting time, pH and calcium release of gray mineral trioxide aggregate (GMTA). Materials and Methods: GMTA powder was mixed with water, 2% CHX alone or 2% CHX combined with 5% CaCl2 . Antimicrobial activity was determined against Enterococcus faecalis (ATCC 29212) strains by agar diffusion test. Data obtained were submitted to kruskal wallis tests. Analysis of the setting time was evaluated by American society for testing and materials C266-03 requirements. The pH and calcium release analysis were evaluated, in 24 h, 7, 14 and 28 days using pH meter equipment and atomic absorption spectrophotometer, respectively. Data obtained were analyzed by ANOVA, in 5% significance level. Results: Significant differences were seen (P < 0.01) among the zones of bacterial growth inhibition produced by 5% CaCl2 + 2% CHX combination against E. faecalis when compared with water (P < 0.05). Regarding the setting time, that combination had the shortest setting time (P < 0.05). All associations were alkaline and released calcium. No statistical difference was observed between the experimental groups at the different periods of analysis (P > 0.05). Conclusion: Combination of 5% CaCl2 + 2% CHX reduced the setting time and enhanced the antimicrobial activity of GMTA without changing the pH and calcium release.