Determinação do período de estabilidade de soluções volumétricas rotineiramente utilizadas em laboratórios de análises químicas / Determination of the stability period of volumetric solutions routinely used in chemical analysis laboratories / Determinación del período de estabilidad de las soluciones volumétricas utilizadas habitualmente en los laboratorios de análisis químicos

As soluções volumétricas são rotineiramente utilizadas nos laboratórios, principalmente nos processos de síntese de produtos e nas análises quantitativas de matéria-prima e/ou produto acabado, entretanto poucos são os estudos que abordam a estabilidade destas soluções. Considerando que a qualidade das soluções volumétricas pode afetar os procedimentos de análises químicas e consequentemente induzir a erros, e ainda que, a Farmacopeia Brasileira (2010) não cita tempo máximo de utilização dessas soluções padronizadas, a avaliação da estabilidade das mesmas é importante. Sendo assim, o objetivo do trabalho foi avaliar a estabilidadede 10 soluções volumétricas, empregadas rotineiramente em laboratórios de análises químicas, com o intuito de estabelecer o período que essas soluções permanecem estáveis, isto é, sem sofrer alteração na concentração. As metodologias de preparo e padronização das soluções volumétricas seguiram os métodos descritos na Farmacopeia Brasileira (2010), sendo as mesmas padronizadas no momento do preparo e a cada 20 dias, por um período de 180 dias. As soluções contendo ácidos e bases, bem como as soluções de iodato de potássio e nitrato de prata, permaneceram constantes durante o período de análises. As soluções de EDTA, iodo, nitrito de sódio, permanganato de potássio e tiossulfato de sódio apresentaram estabilidade inferior a 180 dias, tornando necessária a realização de padronização periódica. As soluções volumétricas utilizadas nos laboratórios apresentam diferentes estabilidades, o que ressalta a importância da determinação do período que as mesmas se mantêmcom as concentrações estáveis, evitando possíveis alterações de resultados nas análises químicas.

Volumetric solutions are routinely used in laboratories, mainly in product synthesis processes and in quantitative analyzes of raw materials and/or finished products, however there are few studies that address the stability of these solutions. Considering that the quality of volumetric solutions can affect chemical analysis procedures and consequently induce errors, and even though the Brazilian Pharmacopoeia (2010) does not mention the maximum time for using these standardized solutions, the evaluation of their stability is important. Therefore, the aim of this work was to evaluate the stability of 10 volumetric solutions, routinely used in chemical analysis laboratories, in order to establish the period that these solutions remain stable without changing their concentrations. The methodologies for preparing and standardizing the volumetric solutions followed the methods described in the Brazilian Pharmacopoeia (2010), being standardized at the time of preparation and every 20 days, for a period of 180 days. Solutions containing acids and bases, as well as potassium iodate and silver nitrate solutions, were stable during the analysis period. The solutions of EDTA, iodine, sodium nitrite, potassium permanganate and sodium thiosulfate showed stability less than 180 days, making it necessary to carry out periodic standardization of these solutions. The volumetric solutions used in the laboratories have different stabilities, which highlights the importance of determining the period in which they remain stable, avoiding possible changes in results in chemical analyzes.

Las soluciones volumétricas se utilizan de forma rutinaria en los laboratorios, principalmente en los procesos de síntesis de productos y en el análisis cuantitativo de materias primas y/o productos acabados. Sin embargo, existen pocos estudios que aborden la estabilidad de estas soluciones. Considerando que la calidad de las soluciones volumétricas puede afectar los procedimientos de análisis químico y consecuentemente inducir a errores, y también que, la Farmacopea Brasileña (2010) no menciona el tiempo máximo de uso de estas soluciones estandarizadas, la evaluación de su estabilidad es importante. Así, el objetivo del trabajo fue evaluar la estabilidad de 10 soluciones volumétricas, utilizadas rutinariamente en los laboratorios de análisis químico, con el fin de establecer el período en que estas soluciones permanecen estables, es decir, sin sufrir alteraciones en la concentración. Las metodologías de preparación y estandarización de las soluciones volumétricas siguieron los métodos descritos en la Farmacopea Brasileña (2010), siendo las mismas estandarizadas en el momento de la preparación y cada 20 días, por un período de 180 días. Las soluciones que contienen ácidos y bases, así como las soluciones de yodato de potasio y nitrato de plata, permanecieron constantes durante el periodo de análisis. Las soluciones de EDTA, yodo, nitrito de sodio, permanganato de potasio y tiosulfato de sodio fueron estables durante menos de 180 días, por lo que fue necesario realizar estandarizaciones periódicas. Las soluciones volumétricas utilizadas en los laboratorios presentan diferentes estabilidades, lo que pone de manifiesto la importancia de determinar el periodo que permanecen con concentraciones estables, evitando posibles cambios en los resultados en los análisis químicos.

Nanofiller Particles and Bonding Durability, Water Sorption, and Solubility of Universal Adhesives.

The aim of this study was to evaluate the influence of nanofiller particles in simplified universal adhesive on the long-term microtensile bond strength and silver nitrate up-take, as well as water sorption and solubility. Commercial adhesives Ambar Universal (FGM) in nanofilled-containing version (filled) and same lot without fillers (unfilled) were donated and applied by means of etch-and-rinse strategy. Microtensile bond strength was surveyed after 24-hours or 1-year water storage. Silver nitrate uptake was assayed using scanning electron microscopy (SEM). Water sorption and solubility experiments were performed based on ISO 4049:2009. Statistical analysis was performed using two-way ANOVA and Tukey test (p<0.05). The bond strength of both the adhesives were statistically similar at 24 hours (p>0.05), but the filled group attained significant bond strength reduction after aging when compared to initial bond strength (p<0.001). Conversely, unfilled adhesive presented stable adhesion after 1-year storage (p=0.262). Silver nitrate uptake was similar for both adhesives, with little silver impregnation at the hybrid and adhesive layers. Water sorption was higher with filled adhesive compared to the unfilled one (p=0.01). Conversely, solubility was higher in unfilled in comparison to filled one (p=0.008). The presence of nanofillers in universal adhesive achieves higher water sorption and dentin bond degradation, which did not occur in the unfilled adhesive.

Oral bioaccessibility of silver nanoparticles and ions in natural soils: Importance of soil properties.

The abundance of silver nanoparticles (AgNPs) in consumer products has led to their environmental release and therefore to concern about their impact on human health. The ingestion of AgNP-contaminated soil from urban sites is an important exposure pathway, especially for children. Given the limited information on oral bioaccessibility of soil Ag, we used a physiologically based extraction test (PBET) to evaluate the bioaccessibility of AgNPs and AgNO3 from soil digestion. The AgNPs underwent several biochemical transformations, including their simultaneous dissolution and agglomeration in gastric fluid followed by the disintegration in the intestinal fluid of the agglomerates into NPs containing silver and chlorine. Therefore, Ag-containing soil exposed the intestine to nanoparticulate Ag in forms that were structurally different from the original forms. The bioaccessibility of AgNPs (0.5 ±â€¯0.05%-10.9 ±â€¯0.7%) was significantly lower than that of AgNO3 (4.7 ±â€¯0.6%-14.4 ±â€¯0.1%), as a result of the lower adsorption of nanoparticles to soil residues during the digestive process. For the soils tested, the bioaccessibility of AgNPs increased with decreasing clay contents and lower pH. By identifying the soil properties that control AgNP bioaccessibility, a more efficient and accurate screening can be performed of soil types that pose the greatest health risk associated with AgNP exposure.

Partitioning of nanoparticle-originated dissolved silver in natural and artificial sediments.

Sediments are believed to be a major sink for silver nanoparticles (AgNPs) in the aquatic environment, but there is a lack of knowledge about the environmental effects and behavior of AgNPs in sediments. The release of highly toxic Ag+ through dissolution of AgNPs is one mechanism leading to toxic effects in sediments. We applied an ultrasound-assisted sequential extraction method to evaluate the dissolution of AgNPs and to study the partitioning of dissolved Ag in sediments. Silver was spiked into artificial and 2 natural sediments (Lake Höytiäinen sediment and Lake Kuorinka sediment) as silver nitrate (AgNO3 ), uncoated AgNPs, or polyvinylpyrrolidone-coated AgNPs (PVP-AgNPs). In addition, the total body burdens of Ag in the sediment-dwelling oligochaete Lumbriculus variegatus were assessed over a 28-d exposure period. The dissolution rate was found to be similar between the uncoated AgNP and PVP-AgNP groups. In all sediments, dissolved Ag was mainly bound to the residual fraction of the sediment, followed by iron and manganese oxides or natural organic matter. In Lake Kuorinka sediment, dissolved Ag that originated from PVP-AgNPs was relatively more bioaccessible, also resulting in higher total body burden in L. variegatus than that from uncoated AgNPs or AgNO3 . In artificial sediment and Lake Höytiäinen sediment, AgNO3 was significantly more bioaccessible than AgNPs. Our results highlight the importance of sediment properties and AgNP surface chemistry when evaluating the environmental exposure of AgNPs. Environ Toxicol Chem 2017;36:2593-2601. © 2017 SETAC.

Silver nanoparticles induce oocyte maturation in zebrafish (Danio rerio).

Public concern regarding silver nanoparticles (AgNPs) in the environment has been increasing since they can cause adverse effects in some aquatic species. However, few data are actually available on the effects of AgNPs on the germ cells. In the present study, we used the zebrafish ovarian follicle as a model to assess the potentially adverse effects of AgNPs on oocyte maturation (germinal vesicle breakdown, GVBD) in vitro. Similar to the maturation inducing hormone (17α, 20ß-dihydroxy-4-pregnen-3-one), AgNPs induced GVBD, and reduced the total cyclic adenosine monophosphate (cAMP) concentration in zebrafish ovarian follicles. The results from transmission electron microscope observation and Hoechst 33342 staining clearly indicated that AgNPs induced apoptosis in ovarian follicle cells surrounding the oocyte. Similar to AgNPs, AgNO3 also induced GVBD, decreased cAMP concentration and induced apoptosis of ovarian follicle cells. However, the results from gene expression analysis showed that transcript levels of oxidative stress related genes were more sensitive to AgNPs than AgNO3. Further more, H2O2 has an ability to induce zebrafish oocytes maturation by induction of apoptosis in ovarian follicle cells. Taken together, the results from our study indicated that oxidative stress appeared to be one of important mechanisms in AgNP induced apoptosis in ovarian follicle cells, which further triggered the GVBD.

Tracking silver delivery to bacteria using turn-on fluorescence.

Herein we present a novel silver complex [Ag(qBODIPY)(CF3SO3)] (1) that exhibits dramatic increase in fluorescence upon silver release. Complex 1 has a minimum inhibitory concentration comparable to that of silver nitrate. Confocal microscopy was used to track the delivery of silver to bacterial targets.

A Filter-based Surface Enhanced Raman Spectroscopic Assay for Rapid Detection of Chemical Contaminants.

We demonstrate a method to fabricate highly sensitive surface-enhanced Raman spectroscopic (SERS) substrates using a filter syringe system that can be applied to the detection of various chemical contaminants. Silver nanoparticles (Ag NPs) are synthesized via reduction of silver nitrate by sodium citrate. Then the NPs are aggregated by sodium chloride to form nanoclusters that could be trapped in the pores of the filter membrane. A syringe is connected to the filter holder, with a filter membrane inside. By loading the nanoclusters into the syringe and passing through the membrane, the liquid goes through the membrane but not the nanoclusters, forming a SERS-active membrane. When testing the analyte, the liquid sample is loaded into the syringe and flowed through the Ag NPs coated membrane. The analyte binds and concentrates on the Ag NPs coated membrane. Then the membrane is detached from the filter holder, air dried and measured by a Raman instrument. Here we present the study of the volume effect of Ag NPs and sample on the detection sensitivity as well as the detection of 10 ppb ferbam and 1 ppm ampicillin using the developed assay.

Harmful effects of silver nanoparticles on a complex detrital model system.

The rapid proliferation of silver nanoparticles (AgNP) in industry and the environment requires realistic toxicity assessments based on approaches that consider the biological complexity of ecosystems. Here we assessed the acute toxicity of carbonate-coated AgNP and, for comparison, AgNO3 (Ag(+)) by using a model system consisting of decomposing plant litter and the associated fungal and bacterial decomposers as central players in the functioning of stream ecosystems. Little variation in size and surface charge during the experiment indicated that the AgNP used were essentially stable. AgNP disrupted bacterial growth (≤83% reduction in protein biosynthesis, EC50 = 0.3 µM), clearly affected fungal growth (≤61% reduction in ergosterol synthesis, EC50 = 47 µM) with both endpoints more sensitive to AgNP than to Ag(+). Fungal reproduction, in contrast, was stimulated by AgNP, but not Ag(+), at concentrations up to 25 µM. Both AgNP and Ag(+ )also stimulated extracellular alkaline phosphatase but reduced leucine aminopeptidase, whereas ß-glucosidase was stimulated by AgNP and reduced by Ag(+). Importantly, the provision of cysteine, a chelating ligand that complexes free Ag(+), failed to alleviate AgNP toxicity to microbial growth, clearly demonstrating particle-mediated toxicity independent of the presence of ionic silver. This contrasts with the observed inhibition of leucine aminopeptidase by Ag(+), which accounted for 2-6% of the total silver in treatments receiving AgNP. These results show that although outcomes of AgNP and Ag(+ )exposure assessed by different functional endpoints vary widely, AgNP strongly interferes with bacterial growth and a range of other microbial processes, resulting in severe consequences for natural microbial communities and ecosystem functioning.

Distribution and accumulation of 10 nm silver nanoparticles in maternal tissues and visceral yolk sac of pregnant mice, and a potential effect on embryo growth.

We examined the distribution of silver in pregnant mice and embryos/fetuses following intravenous injections of 10 nm silver nanoparticles (AgNPs) or soluble silver nitrate (AgNO3) at dose levels of 0 (citrate buffer control) or 66 µg Ag/mouse to pregnant mice on gestation days (GDs) 7, 8 and 9. Selected maternal tissues and all embryos/fetuses from control, AgNP- and AgNO3-treated groups on GD10 and control and AgNP-treated groups on GD16 were processed for the measurement of silver concentrations, intracellular AgNP localization, histopathology and gross examination of tissue morphology. Inductively-coupled plasma mass spectrometry revealed silver in all examined tissues following either AgNP or AgNO3 treatment, with highest concentrations of silver in maternal liver, spleen and visceral yolk sac (VYS), and lowest concentrations in embryos/fetuses. For VYS, mean silver concentration following AgNO3 treatment (4.87 ng Ag/mg tissue) was approximately two-fold that following AgNP treatment (2.31 ng Ag/mg tissue); for all other tissues examined, mean silver concentrations following either AgNP or AgNO3 treatment were not significantly different from each other (e.g. 2.57 or 2.84 ng Ag/mg tissue in maternal liver and 1.61 or 2.50 ng Ag/mg tissue in maternal spleen following AgNP or AgNO3 treatment, respectively). Hyperspectral imaging revealed AgNP aggregates in maternal liver, kidney, spleen and VYS from AgNP-treated mice, but not AgNO3-treated mice. Additionally, one or more embryos collected on GD10 from eight of ten AgNP-treated mice appeared small for their age (i.e. Theiler stage 13 [GD8.5] or younger). In the control group (N = 11), this effect was seen in embryos from only one mouse. In conclusion, intravenous injection of 10 nm AgNPs to pregnant mice resulted in notable silver accumulation in maternal liver, spleen and VYS, and may have affected embryonic growth. Silver accumulation in embryos/fetuses was negligible.

Effects of soil and dietary exposures to Ag nanoparticles and AgNO3 in the terrestrial isopod Porcellionides pruinosus.

The effects of Ag-NPs and AgNO3 on the isopod Porcellionides pruinosus were determined upon soil and dietary exposures. Isopods avoided Ag in soil, with EC50 values of ∼16.0 and 14.0 mg Ag/kg for Ag-NPs and AgNO3, respectively. Feeding inhibition tests in soil showed EC50s for effects on consumption ratio of 127 and 56.7 mg Ag/kg, respectively. Although similar EC50s for effects on biomass were observed for nanoparticulate and ionic Ag (114 and 120 mg Ag/kg dry soil, respectively), at higher concentrations greater biomass loss was found for AgNO3. Upon dietary exposure, AgNO3 was more toxic, with EC50 for effects on biomass change being >1500 and 233 mg Ag/kg for Ag-NPs and AgNO3, respectively. The difference in toxicity between Ag-NPs and AgNO3 could not be explained from Ag body concentrations. This suggests that the relation between toxicity and bioavailability of Ag-NPs differs from that of ionic Ag in soils.

Development and application of methods for the determination of silver in polymeric dressings used for the care of burns.

Open vessel and microwave digestion methods have been developed for the determination of total silver in six commercial dressing used for the treatment of skin burns. An extraction method using TMAH has also been developed to determine the amount of silver present in the exudates found on the surface after dressing removal so an estimation of the patient dose can be made. All microwave methods had a quantitative recovery, whereas the open vessel had recoveries that ranged from 80 to 100%. The silver concentrations were determined by inductively coupled plasma mass spectrometry using an external calibration. In the absence of suitable reference materials, isotope dilution analysis was applied to validate the accuracy of results obtained by external calibration. All the products had a total Ag content that agreed with the values declared by the producer, which ranged from 10 to 0.2% Ag by weight. One of the methods was applied to the indirect determination of Ag released in vivo by Acticoat™ Flex 3, a dressing composed of silver nanoparticles on a polymer net. Silver levels were determined in used dressings after application to patients with partial thickness skin burns. A maximum of 62% of the silver was found to have been released onto the patient where hemopurulent exudate occurred, indicating that the dressing was virtually exhausted after 3 days of use. We conclude that the Ag released into the patient's tissues is closely correlated with the local severity of the wound.

Behaviour of silver nanoparticles and silver ions in an in vitro human gastrointestinal digestion model.

Oral ingestion is an important exposure route for silver nanoparticles (AgNPs), but their fate during gastrointestinal digestion is unknown. This was studied for 60 nm AgNPs and silver ions (AgNO3) using in vitro human digestion model. Samples after saliva, gastric and intestinal digestion were analysed with SP-ICPMS, DLS and SEM-EDX. In presence of proteins, after gastric digestion the number of particles dropped significantly, to rise back to original values after the intestinal digestion. SEM-EDX revealed that reduction in number of particles was caused by their clustering. These clusters were composed of AgNPs and chlorine. During intestinal digestion, these clusters disintegrated back into single 60 nm AgNPs. The authors conclude that these AgNPs under physiological conditions can reach the intestinal wall in their initial size and composition. Importantly, intestinal digestion of AgNO3 in presence of proteins resulted in particle formation. These nanoparticles (of 20-30 nm) were composed of silver, sulphur and chlorine.

Aging and soil organic matter content affect the fate of silver nanoparticles in soil.

Sewage sludge application on soils represents an important potential source of silver nanoparticles (Ag NPs) to terrestrial ecosystems, and it is thus important to understand the fate of Ag NPs once in contact with soil components. Our aim was to compare the behavior of three different forms of silver, namely silver nitrate, citrate stabilized Ag NPs (5nm) and uncoated Ag NPs (19nm), in two soils with contrasting organic matter content, and to follow changes in binding strength over time. Soil samples were spiked with silver and left to age for 2h, 2 days, 5 weeks or 10 weeks before they were submitted to sequential extraction. The ionic silver solution and the two Ag NP types were radiolabeled so that silver could be quantified by gamma spectrometry by measuring the (110m)Ag tracer in the different sequential extraction fractions. Different patterns of partitioning of silver were observed for the three forms of silver. All types of silver were more mobile in the mineral soil than in the soil rich in organic matter, although the fractionation patterns were very different for the three silver forms in both cases. Over 20% of citrate stabilized Ag NPs was extractible with water in both soils the first two days after spiking (compared to 1-3% for AgNO(3) and uncoated Ag NPs), but the fraction decreased to trace levels thereafter. Regarding the 19nm uncoated Ag NPs, 80% was not extractible at all, but contrary to AgNO(3) and citrate stabilized Ag NPs, the bioaccessible fraction increased over time, and by day 70 was between 8 and 9 times greater than that seen in the other two treatments. This new and unexpected finding demonstrates that some Ag NPs can act as a continuous source of bioaccessible Ag, while AgNO(3) is rapidly immobilized in soil.

In vitro biosynthesis and genotoxicity bioassay of silver nanoparticles using plants.

Silver nanoparticles (AgNP-P) from AgNO(3) were synthesized by using the broth prepared from the aromatic spath of male inflorescence of screw pine, Pandanus odorifer (Forssk.) Kuntze AgNP-P was then characterized by UV-visible spectroscopy, transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDS). Functional groups in the broth were analyzed by Fourier Transform infrared spectroscopy (FTIR). Genotoxicity of AgNP-P was assessed by utilizing our well-established Allium cepa assay system with biomarkers including the generation reactive oxygen species (ROS: O(2)(·-) and H(2)O(2)), cell death, mitotic index, micronucleus, mitotic aberrations; and DNA damage by Comet assay. Other chemical forms of silver such as Ag(+) ion, colloidal AgCl, and AgNP-S at doses 0-80 mg L(-1) were included for comparison with AgNP-P. The results revealed that AgNP-P and AgNP-S exhibited similar biological effects in causing lesser extent of cytotoxicity and greater extent of genotoxicity than that was exhibited by Ag(+) ion alone. Among different tested chemical forms of silver, colloidal AgCl was identified to be the least cytotoxic and genotoxic. Cell death and DNA-damage induced by AgNP-P were prevented by Tiron and dimethyl thiourea that scavenge O(2)(·-) and H(2)O(2), respectively. The present findings demonstrated the role of ROS in the AgNP-induced cell death and DNA damage.

Biocompatibility and antibacterial effect of silver doped 3D-glass-ceramic scaffolds for bone grafting.

A 3D-glass-ceramic scaffold for bone tissue engineering with an interconnected macroporous network of pores was doped with silver ions in order to confer antibacterial properties. For this purpose, silver ions were selectively added to the scaffold surfaces through ion-exchange using an aqueous silver nitrate solution. The silver-doped scaffolds were characterized by means of leaching, in vitro antibacterial, and citotoxicity tests. In particular, the silver effect was examined through a broth dilution test in order to evaluate the proliferation of bacteria by counting the colonies forming units. Moreover, cytotoxicity tests were carried out to understand the effect of silver-containing scaffolds on cell adhesion, proliferation, and vitality. For all tests a comparison between silver-doped scaffold and silver-doped scaffold dry sterilized was performed.

Mechanism of bactericidal activity of Silver Nitrate: a concentration dependent bi-functional molecule

Silver nitrate imparts different functions on bacteria depending upon its concentration. At lower concentration it induced synthesis of nanoparticles, whereas at higher concentrations it induced cell death. Bacillus licheniformis was used as model system. The MIC was 5 mM, and it induced catalase production, apoptotic body formation and DNA fragmentation.

Effects of adhesive temperature on the early and 6-month dentin bonding.

OBJECTIVES: The aim of this study was to test the effect of adhesive temperature on the bond strength to dentin (muTBS) and silver nitrate uptake (SNU) of an ethanol/water (Adper Single Bond 2 [SB]) and an acetone-based (Prime&Bond 2.1 [PB]) etch-and-rinse adhesive system. METHODS: The bottles of each adhesive were kept in various temperatures (5 degrees C, 20 degrees C, 37 degrees C and 50 degrees C) for 1h previously to its application in the occlusal demineralized dentin of 40 molars. Bonded sticks (0.8 mm(2)) were tested in tension (0.5 mm/min) immediately (IM) or after 6 months (6 M) of water storage. Two bonded sticks from each hemi-tooth were immersed in silver nitrate and analyzed by SEM. Data were analyzed by two-way repeated measures ANOVA and Tukey's test (alpha=0.05). RESULTS: No significant difference in muTBS was detected for both adhesives at 5 degrees C and 20 degrees C. The highest bond strength for PB was observed in the 37 degrees C group while for SB it was in the 50 degrees C. Significant reductions of bond strengths were observed for PB at 37 degrees C and SB at 50 degrees C after 6 M of water storage. Silver nitrate deposition was seen in all hybrid layers, irrespective of the group. Lower silver nitrate deposition (water trees) in the adhesive layer was seen for PB and SB at higher temperatures. CONCLUSIONS: The heating or refrigeration of the adhesives did not improve their resin-dentin bond resistance to water degradation over time.

The in vitro reduction of sodium [36Cl]chlorate in bovine ruminal fluid.

Sodium chlorate effectively reduces or eliminates gram-negative pathogenic bacteria in the gastrointestinal tracts of live cattle. Limitations to the in vivo efficacy of chlorate are its rapid absorption from the gastrointestinal tract and its presumed reduction to chloride within the gastrointestinal tract. We hypothesized that chlorate would be reduced via ruminal bacteria in a ruminal in vitro system and that the reduction of chlorate would be influenced by the dietary for-age:concentrate ratio; thus, 4 ruminally cannulated steers were fed 20 or 80% concentrate diets in a crossover design. Ruminal fluid was collected in 2 periods and dispensed into in vitro tubes containing sodium [36Cl]chlorate, which was sufficient for 100 or 300 mg/L final chlorate concentrations. The tubes were incubated for 0, 1, 4, 8, 16, or 24 h; autoclaved, control ruminal fluid, fortified with sodium [36Cl]chlorate, was incubated for 24 h. Chlorate remaining in each sample was measured by liquid scintillation counting after [36Cl]chloride was precipitated with silver nitrate. A preliminary study indicated that chlorite, a possible intermediate in the reduction of chlorate, had a half-life of approximately 4.5 min in freshly collected (live) ruminal fluid; chlorite was, therefore, not specifically measured in ruminal incubations. The chlorate dose did not affect in vitro DM digestion (P > or = 0.11), whereas in vitro DM digestibility was decreased (P < or = 0.05) by 80% forage content. By 24 h, 57.5 +/- 2.6% of the chlorate remained in 100-mg/L incubations, whereas 78.2 +/- 2.6% of the chlorate remained in the 300-mg/L incubations. When the data were expressed on a concentration basis (mg/L), diet had no effect (P > or = 0.18) on chlorate reduction; however, when chlorate reduction was expressed on a percentage basis, chlorate reduction tended to be greater (P > or = 0.09) at 8 and 16 h in the incubations containing the low-concentrate diet. Chlorate remaining in autoclaved controls at 24 h was intermediate (P < 0.01) between chlorate remaining in live ruminal fluid samples incubated for 0 or 24 h. Attempts to isolate chlorate-respiring bacteria from 2 sources of ruminal fluid were not successful. These data indicate that microbial-dependent or chemical-dependent, or both, reduction of chlorate occurs in bovine ruminal fluid and that dietary concentrate had a negligible effect on chlorate reduction.

Dentin penetrability evaluation of three different dyes in root-end cavities filled with mineral trioxide aggregate (MTA).

The purpose of this study was to evaluate the penetration of three dyes in MTA root-end fillings. In 30 single-rooted teeth, cavities for retrofilling were prepared with an ultrasound appliance and filled with MTA. The specimens were randomly assigned to three groups (n = 10) and immersed in the following solutions: 2% methylene blue (MET), 50% silver nitrate (NIT) and 0.2% rhodamine B (ROD). Two transversal slices (1 mm) of the retrofilling region were obtained and evaluated using the Image Tool 3.0 software to obtain a quantitative evaluation (in mm2) of the dye penetration around the retrofillings. Data were submitted to statistical analysis using Students t-test. The lowest degree of dye penetration was observed for the NIT group, in both slices (p < 0.05). Dye penetration was significantly larger in the ROD group when compared to the NIT group, in both slices (p < 0.05), and to the MET group, only in slice 1 (p < 0.05). Within the limitations of this study, it was concluded that the choice of dye could influence the penetration evaluation in root-end filling studies, and that the NIT had the lowest penetration capacity in the apical dentine.

Dentin penetrability evaluation of three different dyes in root-end cavities filled with mineral trioxide aggregate (MTA)

A proposta deste trabalho foi avaliar a penetração de três corantes em retrocavidades obturadas com MTA. As retrocavidades foram confeccionadas com aparelho de ultra-som em 30 dentes unirradiculares e obturadas com MTA. Os espécimes foram divididos aleatoriamente em três grupos (n = 10) e imersos nas seguintes soluções corantes: azul de metileno a 2% (MET), nitrato de prata a 50% (NIT) e rodamina B a 0,2% (ROD). Duas fatias transversais (1 mm) da região retrobturada foram obtidas e avaliadas através do software Image Tool 3.0, objetivando quantificar a área (em mm2) de penetração do corante ao redor das retrobturações. Os dados foram submetidos à análise estatística, utilizando o teste t de Student. A menor área de penetração foi observada no grupo NIT, nas duas fatias (p < 0.05). A penetração de corante foi significativamente maior no grupo ROD quando comparado ao grupo NIT, nas duas fatias (p < 0.05), e ao grupo MET, somente na fatia 1 (p < 0.05). Dentro das limitações desta pesquisa, concluiu-se que a escolha da solução corante pode influenciar a avaliação da penetração em estudos sobre retrobturações e que o grupo NIT teve a menor capacidade de penetração na dentina apical.