Simplified vacuum dressing system: effectiveness and safety in wounds management

Purpose: Negative pressure wound therapy (NPWT) has revolutionized wound care, but its high cost reduces the procedure's availability. To solve the problem, streamlined vacuum dressings systems have been proposed, but the utility of these devices has been poorly studied. The objective of this study was to evaluate a simplified vacuum dressing system model (SVDM). Methods: Randomized clinical trial in which wounds were treated with SVDM compared to a complex occlusive dressing (silver hydrofiber, SHF). The analyzed outcomes were cleaning, presence of granulation tissue, clinical appearance, and indication for surgical closure of wounds. Results: Fifty injuries were treated (25 in each group), most located on lower limbs. SVDM proved to be more effective than SHF in the evaluated outcomes. Wound recalcitrance reduced the effectiveness of the equipment used. Despite its efficacy, complications occurred, the most frequent related to dressing changes: minor bleeding, foam adherence to a wound bed, and pain. Only for bleeding no favorable risk-benefit ratio was found. There were no severe complications, worsening conditions of injuries, or deaths. Conclusions: SVDM proved to be an effective and acceptably safe device for managing studied wounds.

Speciation Analysis of Ag2S and ZnS Nanoparticles at the ng/L Level in Environmental Waters by Cloud Point Extraction Coupled with LC-ICPMS.

Toxicity and transport of metal-based nanoparticles (M-NPs) in environmental waters strongly depend on their speciation. A detailed understanding of the composition and speciation of M-NPs is necessary in order to move this field forward. Unfortunately, there is a shortage of analytical methods for metal-sulfide nanoparticles (MS-NPs) in the environment. In this work, a cloud point extraction (CPE) method combined with liquid chromatography hyphenated to inductively coupled plasma mass spectrometry (LC-ICPMS) is developed for sensitive determination of Ag2S- and ZnS-NPs. Under the condition of 0.15% (w/v) of Triton X-114 (TX-114), pH 5, 20 mM NaNO3, incubation temperature of 45 °C, and time of 15 min, MS-NPs and non-MS-NPs were extracted into the surfactant-rich phase. With the sequent addition of 10 mM bis(p-sulfonatophenyl)phenylphosphane dehydrate dipotassium (BSPP) aqueous solution (100 µL) into the CPE-obtained extract, the non-MS-NPs were selectively dissociated into their ionic counterparts while maintaining the original size and shape of Ag2S- and ZnS-NPs. Interestingly, the micelle-mediated behavior suddenly disappeared with the addition of BSPP. Thus, the extract can be injected to LC-ICPMS for speciation analysis of trace Ag2S- and ZnS-NPs. This method exhibited excellent reproducibility (relative standard deviations < 4.9%), high sensitivity with the respective detection limits of 8 ng/L for Ag2S-NPs and 15 ng/L for ZnS-NPs, enabling recoveries of 81.3-96.6% for Ag2S-NPs and 83.9-93.5% for ZnS-NPs when they were spiked into three environmental water samples. Due to its potential applicability to low concentrations of Ag2S- and ZnS-NPs, this method is particularly convenient for monitoring the transformations of AgNPs and ZnO-NPs in the environment.

Silver nanoparticles in sewage sludge: Bioavailability of sulfidized silver to the terrestrial isopod Porcellio scaber.

Silver nanoparticles (AgNPs) are efficiently converted during the wastewater-treatment process into sparingly soluble Ag sulfides (Ag2 S). In several countries, sewage sludge is used as a fertilizer in agriculture. The bioavailability of sulfidized Ag to the terrestrial isopod Porcellio scaber was investigated. Sewage sludge containing transformed AgNPs was obtained from a laboratory-scale sewage-treatment plant operated according to Organisation for Economic Co-operation and Development (OECD) guideline 303a. The results of transmission electron microscopy with energy dispersive X-ray of sludge samples suggest that AgNPs were completely transformed to Ag2 S. Adult isopods were exposed to OECD 207 soil substrate amended with the AgNP spiked sludge for 14 d (uptake phase) followed by an elimination phase in unspiked soil of equal duration. Most of the Ag measured in P. scaber at the end of the uptake phase was found in the hindgut (71%), indicating that only a minor part of the estimated Ag content was actually assimilated by the isopods with 16.3 and 12.7% found in the carcass and hepatopancreas, respectively. As a result of this, the Ag content of the animals dropped following transition to unspiked sludge within 2 d to one-third of the previously measured Ag concentration and remained stable at this level until the end of the elimination period. The present study shows that Ag2 S in sewage sludge is bioavailable to the terrestrial isopod P. scaber. Environ Toxicol Chem 2018;37:1606-1613. © 2018 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Long-term effects of sulfidized silver nanoparticles in sewage sludge on soil microflora.

The use of silver nanoparticles (AgNPs) in consumer products such as textiles leads to their discharge into wastewater and consequently to a transfer of the AgNPs to soil ecosystems via biosolids used as fertilizer. In urban wastewater systems (e.g., sewer, wastewater treatment plant [WWTP], anaerobic digesters) AgNPs are efficiently converted into sparingly soluble silver sulfides (Ag2 S), mitigating the toxicity of the AgNPs. However, long-term studies on the bioavailability and effects of sulfidized AgNPs on soil microorganisms are lacking. Thus we investigated the bioavailability and long-term effects of AgNPs (spiked in a laboratory WWTP) on soil microorganisms. Before mixing the biosolids into soil, the sludges were either anaerobically digested or directly dewatered. The effects on the ammonium oxidation process were investigated over 140 d. Transmission electron microscopy (TEM) suggested an almost complete sulfidation of the AgNPs analyzed in all biosolid samples and in soil, with Ag2 S predominantly detected in long-term incubation experiments. However, despite the sulfidation of the AgNPs, soil ammonium oxidation was significantly inhibited, and the degree of inhibition was independent of the sludge treatment. The results revealed that AgNPs sulfidized under environmentally relevant conditions were still bioavailable to soil microorganisms. Consequently, Ag2 S may exhibit toxic effects over the long term rather than the short term. Environ Toxicol Chem 2017;36:3305-3313. © 2017 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Stability of single dispersed silver nanoparticles in natural and synthetic freshwaters: Effects of dissolved oxygen.

Silver nanoparticles (AgNPs) are increasingly used in various commercial products. This increased use raises ecological concerns because of the large release of AgNPs into the environment. Once released, the local water chemistry has the potential to influence the environmental fates and behaviors of AgNPs. The impacts of dissolved oxygen and natural organic matter (NOM) on the dissolution and stability of AgNPs were investigated in synthetic and natural freshwaters for 7 days. In synthetic freshwater, the aggregation of AgNPs occurred due to the compression of the electric double layer, accompanied by the dissolution of AgNPs. However, once oxygen was removed, the highest dissolved Ag (Agdis) concentration decreased from 356.5 µg/L to 272.1 µg/L, the pH of the AgNP suspensions increased from less than 7.6 to more than 8.4, and AgNPs were regenerated by the reduction of released Ag+ by citrate. The addition of NOM mitigated aggregation, inhibited oxidative dissolution and induced the transformation of AgNPs into Ag2S due to the formation of NOM-adsorbed layers, the reduction of Ag+ by NOM, and the high affinity of sulfur-enriched species in NOM for Ag. Likewise, in oxygen-depleted natural freshwaters, the inhibition of oxidative dissolution was obtained in comparison with oxygenated freshwaters, showing a decrease in the maximum Agdis concentration from 137.6 and 57.0 µg/L to 83.3 and 42.4 µg/L from two natural freshwater sites. Our results suggested that aggregation and dissolution of AgNPs in aquatic environments depend on the chemical composition, where oxygen-depleted freshwaters more significantly increase the colloidal stability. In comparison with oxic conditions, anoxic conditions were more favorable to the regeneration of AgNPs by reducing species (e.g., citrate and NOM) and enhanced the stability of nanoparticles. This indicates that some AgNPs will be more stable for long periods in oxygen-deprived freshwaters, and pose more serious environmental risks than that in oxygenated freshwaters.

The effect of chronic silver nanoparticles on aquatic system in microcosms.

Silver nanoparticles (AgNPs) inevitably discharge into aquatic environments due to their abundant use in antibacterial products. It was reported that in laboratory conditions, AgNPs display dose-dependent toxicity to aquatic organisms, such as bacteria, algae, macrophytes, snails and fishes. However, AgNPs could behave differently in natural complex environments. In the present study, a series of microcosms were established to investigate the distribution and toxicity of AgNPs at approximately 500 µg L-1 in aquatic systems. As a comparison, the distribution and toxicity of the same concentration of AgNO3 were also determined. The results showed that the surface layer of sediment was the main sink of Ag element for both AgNPs and AgNO3. Both aquatic plant (Hydrilla verticillata) and animals (Gambusia affinis and Radix spp) significantly accumulated Ag. With short-term treatment, phytoplankton biomass was affected by AgNO3 but not by AgNPs. Chlorophyll content of H. verticillata increased with both AgNPs and AgNO3 short-term exposure. However, the biomass of phytoplankton, aquatic plant and animals was not significantly different between control and samples treated with AgNPs or AgNO3 for 90 d. The communities, diversity and richness of microbes were not significantly affected by AgNPs and AgNO3; in contrast, the nitrification rate and its related microbe (Nitrospira) abundance significantly decreased. AgNPs and AgNO3 may affect the nitrogen cycle and affect the environment and, since they might be also transferred to food web, they represent a risk for health.

Symbiosis between nitrogen-fixing bacteria and Medicago truncatula is not significantly affected by silver and silver sulfide nanomaterials.

Silver (Ag) engineered nanomaterials (ENMs) are being released into waste streams and are being discharged, largely as Ag2S aged-ENMs (a-ENMs), into agroecosystems receiving biosolids amendments. Recent research has demonstrated that biosolids containing an environmentally relevant mixture of ZnO, TiO2, and Ag ENMs and their transformation products, including Ag2S a-ENMs, disrupted the symbiosis between nitrogen-fixing bacteria and legumes. However, this study was unable to unequivocally determine which ENM or combination of ENMs and a-ENMs was responsible for the observed inhibition. Here, we examined further the effects of polyvinylpyrollidone (PVP) coated pristine Ag ENMs (PVP-Ag), Ag2S a-ENMs, and soluble Ag (as AgSO4) at 1, 10, and 100 mg Ag kg(-1) on the symbiosis between the legume Medicago truncatula and the nitrogen-fixing bacterium, Sinorhizobium melliloti in biosolids-amended soil. Nodulation frequency, nodule function, glutathione reductase production, and biomass were not significantly affected by any of the Ag treatments, even at 100 mg kg(-1), a concentration analogous to a worst-case scenario resulting from long-term, repeated biosolids amendments. Our results provide additional evidence that the disruption of the symbiosis between nitrogen-fixing bacteria and legumes in response to a mixture of ENMs in biosolids-amended soil reported previously may not be attributable to Ag ENMs or their transformation end-products. We anticipate these findings will provide clarity to regulators and industry regarding potential unintended consequences to terrestrial ecosystems resulting from of the use of Ag ENMs in consumer products.

General treatment and wound management in hereditary epidermolysis bullosa: indication and experience using silver hydrofiber dressing / Tratamento geral e das feridas na epidermólise bolhosa hereditária: indicação e experiência usando curativo de hidrofibra com prata

Introduction: Hereditary epidermolysis bullosa (EB) is a rare disorder characterized by cutaneomucous fragility, with formation of blisters during minimal trauma. Treatment consists of clinical and nutritional support and management of pain and skin lesions. Silver hydrofiber (Aquacel Ag®) is a type of carboxymethylcellulose fiber dressing with silver that can be used in selected cases of EB. Objective: To review the literature on the general treatment and management of cutaneous lesions in congenital EB and evaluate the indication and experience of using silver hydrofiber dressing. Methods: The review included original articles and systematic reviews published between 2009 and 2014. We also selected two patients with congenital EB treated at the Plastic Surgery Division of Hospital das Clínicas of the Faculty of Medicine of Ribeirão Preto at the University of São Paulo. Results: There is a shortage of scientific evidence related to the treatment of skin lesions in congenital EB, with most recommendations being based on expert opinions. Hydrofiber is indicated in most consensuses for wounds with some exudation and has been shown to be more absorbent than alginate. In our experience, there was apparent improved control of pain, bleeding, and hypothermia with the use of hydrofiber, which has the advantage of not requiring daily changes and can remain on the wound for up to two weeks. Conclusions: The general and lesion treatments in EB are challenging. Hydrofiber with silver is a treatment option for wounds in hereditary EB, without the need for daily dressing changes.

Introdução: Epidermólise bolhosa (EB) hereditária é uma desordem rara caracterizada pela fragilidade cutaneomucosa, com formação de bolhas ao mínimo trauma. O tratamento consiste em suporte clínico, nutricional, manejo da dor e das lesões cutâneas. A hidrofibra com prata (Aquacel Ag®) é um tipo de curativo de fibra de carboximetilcelulose e prata que pode ser utilizada em casos selecionados de EB. Objetivo: Revisão da literatura sobre o tratamento geral e o manejo das lesões cutâneas na EB congênita, além de avaliar a indicação e experiência usando curativo de hidrofibra com prata. Métodos: A revisão incluiu artigos originais e revisões sistemáticas, publicados entre 2009 e 2014. Selecionamos ainda dois pacientes com EB congênita tratados na Divisão de Cirurgia Plástica do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo. Resultados: Há escassez de evidências científicas relacionadas ao tratamento das lesões cutâneas na EB congênita, sendo a maioria das recomendações baseadas em opiniões de especialistas. A hidrofibra está indicada na maioria dos consensos para feridas com alguma exsudação e mostrou-se mais absorvente que o alginato. Em nossa experiência, houve aparente melhor controle da dor, do sangramento e da hipotermia com o uso da hidrofibra, que apresenta a vantagem de não necessitar de trocas diárias, podendo permanecer na ferida por até duas semanas. Conclusões: O tratamento geral e das lesões na EB é um desafio. A hidrofibra com prata é uma opção de tratamento para as feridas na EB hereditária, sem necessidade de trocas diárias de curativo.

Gas-phase intramolecular cyclization of argentinated N-allylbenzamides.

The fragmentations of argentinated N-allylbenzamides have been exhaustively studied through collision-induced dissociation and through deuterium labeling. The intriguing elimination of AgOH is certified as the consequence of intramolecular cyclization between terminal olefin and carbonyl carbon following proton transfer to carbonyl oxygen, rather than simple enolization of amide. Linear free energy correlations and density functional theory (DFT) calculations were performed to understand the competitive relationship between AgOH loss and AgH loss, which results from the 1,2-elimination of α-hydrogen (to the amido nitrogen) with the silver.

Transformation of AgCl nanoparticles in a sewer system--A field study.

Silver nanoparticles (Ag-NP) are increasingly used in consumer products and their release during the use phase may negatively affect aquatic ecosystems. Research efforts, so far, have mainly addressed the application and use of metallic Ag(0)-NP. However, as shown by recent studies on the release of Ag from textiles, other forms of Ag, especially silver chloride (AgCl), are released in much larger quantities than metallic Ag(0). In this field study, we report the release of AgCl-NP from a point source (industrial laundry that applied AgCl-NP during a piloting phase over a period of several months to protect textiles from bacterial regrowth) to the public sewer system and investigate the transformation of Ag during its transport in the sewer system and in the municipal wastewater treatment plant (WWTP). During the study period, the laundry discharged ~85 g of Ag per day, which dominated the Ag loads in the sewer system from the respective catchment (72-95%) and the Ag in the digested WWTP sludge (67%). Combined results from electron microscopy and X-ray absorption spectroscopy revealed that the Ag discharged from the laundry to the sewer consisted of about one third AgCl and two thirds Ag2S, both forms primarily occurring as nanoparticles with diameters<100 nm. During the 800 m transport in the sewer channel to the nearby WWTP, corresponding to a travel time of ~30 min, the remaining AgCl was transformed into nanoparticulate Ag2S. Ag2S-NP also dominated the Ag speciation in the digested sludge. In line with results from earlier studies, the very low Ag concentrations measured in the effluent of the WWTP (<0.5 µg L(-1)) confirmed the very high removal efficiency of Ag from the wastewater stream (>95%).

Speciation and lability of Ag-, AgCl-, and Ag2S-nanoparticles in soil determined by X-ray absorption spectroscopy and diffusive gradients in thin films.

Long-term speciation and lability of silver (Ag-), silver chloride (AgCl-), and silver sulfide nanoparticles (Ag2S-NPs) in soil were studied by X-ray absorption spectroscopy (XAS), and newly developed "nano" Diffusive Gradients in Thin Films (DGT) devices. These nano-DGT devices were designed specifically to avoid confounding effects when measuring element lability in the presence of nanoparticles. The aging profile and stabilities of the three nanoparticles and AgNO3 (ionic Ag) in soil were examined at three different soil pH values over a period of up to 7 months. Transformation of ionic Ag, Ag-NP and AgCl-NPs were dependent on pH. AgCl formation and persistence was observed under acidic conditions, whereas sulfur-bound forms of Ag dominated in neutral to alkaline soils. Ag2S-NPs were found to be very stable under all conditions tested and remained sulfur bound after 7 months of incubation. Ag lability was characteristically low in soils containing Ag2S-NPs. Other forms of Ag were linked to higher DGT-determined lability, and this varied as a function of aging and related speciation changes as determined by XAS. These results clearly indicate that Ag2S-NPs, which are the most environmentally relevant form of Ag that enter soils, are chemically stable and have profoundly low Ag lability over extended periods. This may minimize the long-term risks of Ag toxicity in the soil environment.

Synergistic coupling of fluorescent "turn-off" with spectral overlap modulated FRET for ratiometric Ag+ sensor.

A useful strategy for ratiometric fluorescent detecting of Ag(+) is demonstrated. Upon selective binding of Ag(+) to a BODIPY-porphyrin dyad (1), the synergistic coupling of two functions, namely the suppressing of FRET from BODIPY donor to porphyrin acceptor and the fluorescence quenching of porphyrin acceptor, leads to exceptionally large changes in the intensity ratio of two distinct emissions (F513/F654) which allow for the ratiometric detecting of Ag(+) with excellent sensitivity in solution and living cells.

Remobilisation of silver and silver sulphide nanoparticles in soils.

Manufactured nanoparticles (NPs) present in consumer products could enter soils through re-use of biosolids. Among these NPs are those based on silver (Ag), which are found sulphidised (e.g. silver sulphide, Ag2S) in biosolids. Herein, our aim was to examine the release of retained Ag(0) and Ag2S NPs in soils and biosolids as facilitated by environmentally and agriculturally relevant ligands. Under natural soil conditions, exemplified by potassium nitrate and humic acid experiments, release of Ag retained in soil was limited. The highest total Ag release was facilitated by ligands that simulated root exudates (citrate) or fertilisers (thiosulphate). Released Ag was predominantly present in the colloidal phase (>3 kDa-< 0.45 µm); intact NPs only identified in Ag2S-NP extracts. For biosolids containing nanoparticulate-Ag-S, release was also enhanced by thiosulphate, though mostly as colloidal-Ag - not intact NPs. These results suggest that exposure to NPs as a result of its release from soils or biosolids will be low.

Inhibitory effect of silver diamine fluoride on dentine demineralisation and collagen degradation.

OBJECTIVE: To investigate the inhibitory effects of 38% silver diamine fluoride (SDF) on demineralised dentine. METHODS: Human dentine blocks were demineralised and allocated to four groups: SF, F, S and W. The blocks in group SF received a topical application of 38% SDF solution (253,900ppm Ag, 44,800ppm F), group F received a 10% sodium fluoride solution (44,800ppm F), group S received a 42% silver nitrate solution (253,900ppm Ag) and group W received deionised water (control). They were subjected to pH cycling using demineralisation solution (pH 5) and remineralisation solution (pH 7) for 8 days. The surface morphology, crystal characteristics, lesion depth and collagen matrix degradation of the specimens were investigated by scanning electron microscopy (SEM), X-ray diffraction (XRD), micro-CT testing and spectrophotometry with a hydroxyproline assay. RESULTS: The surface morphology under SEM showed evident demineralisation with exposed collagen in groups S and W, but not in group SF. Clusters of granular spherical grains were observed in the cross-sections of specimens in groups SF and F. XRD revealed precipitates of silver chloride in groups SF and S. The mean lesion depths (±SD) of groups SF, F, S and W were 182 ± 32µm, 204 ± 26µm, 259 ± 42µm and 265 ± 40µm, respectively (SDF, F

State of the art in human risk assessment of silver compounds in consumer products: a conference report on silver and nanosilver held at the BfR in 2012.

In light of the broad spectrum of products containing nanosilver, the harmfulness of nanosilver to human health and the environment was intensively discussed at a conference held in February 2012 at the BfR. The conference agenda covered the aspects of analytics of nanosilver materials, human exposure and toxicology as well as effects on microorganisms and the environment. The discussion recovered major gaps related to commonly agreed guidelines for sample preparation and central analytical techniques. In particular, the characterization of the nanoparticles in complex matrices was regarded as a challenge which might become a pitfall for further innovation and application. Historical and anecdotal records of colloidal silver have been sometimes taken as empirical proof for the general low toxicity of nanosilver. Yet as reported herein, a growing number of animal studies following modern performance standards of toxicity testing have been carried out recently revealing well-characterized adverse effects on different routes of exposure in addition to argyria. Furthermore, recent approaches in exposure assessment were reported. However, consumer exposure scenarios are only starting to be developed and reliable exposure data are still rare. It was further widely agreed on the workshop that the use of silver may lead to the selection of silver resistant bacteria. With respect to its environmental behavior, it was suggested that nanosilver released to wastewater may have negligible ecotoxicological effects. Finally, the presentations and discussion on risk assessment and regulation of nanosilver applications gave insights into different approaches of risk assessment of nanomaterials to be performed under the various regulatory frameworks.

Transformation of four silver/silver chloride nanoparticles during anaerobic treatment of wastewater and post-processing of sewage sludge.

The increasing use of silver (Ag) nanoparticles [containing either elemental Ag (Ag-NPs) or AgCl (AgCl-NPs)] in commercial products such as textiles will most likely result in these materials reaching wastewater treatment plants. Previous studies indicate that a conversion of Ag-NPs to Ag2S is to be expected during wastewater transport/treatment. However, the influence of surface functionality, the nature of the core structure and the effect of post-processing on Ag speciation in sewage sludge/biosolids has not been investigated. This study aims at closing these knowledge gaps using bench scale anaerobic digesters spiked with Ag nitrate, three different types of Ag-NPs, and AgCl-NPs at environmentally realistic concentrations. The results indicate that neither surface functionality nor the different compositions of the NP prevented the formation of Ag2S. Silver sulfides, unlike the sulfides of other metals present in sewage sludge, were stable over a six month period simulating composting/stockpiling.

Silver behaviour along the salinity gradient of the Gironde Estuary.

Dissolved and particulate Ag concentrations (Ag(D) and Ag(P), respectively) were measured in surface water and suspended particulate matter (SPM) along the salinity gradient of the Gironde Estuary, South West France, during three cruises (2008-2009) covering contrasting hydrological conditions, i.e. two cruises during intermediate and one during high freshwater discharge (~740 and ~2,300 m(3)/s). Silver distribution reflected non-conservative behaviour with 60-70 % of Ag(P) in freshwater particles being desorbed by chlorocomplexation. The amount of Ag(P) desorbed was similar to the so-called reactive, potentially bioavailable Ag(P) fraction (60 ± 4 %) extracted from river SPM by 1 M HCl. Both Ag(P) (0.22 ± 0.05 mg/kg) and Ag(P)/Th(P) (0.025-0.028) in the residual fraction of fluvial and estuarine SPM were similar to those in SPM from the estuary mouth and in coastal sediments from the shelf off the Gironde Estuary, indicating that chlorocomplexation desorbs the reactive Ag(P). The data show that desorption of reactive Ag(P) mainly occurs inside the estuary during low and intermediate discharge, whereas expulsion of partially Ag(P)-depleted SPM (Ag(P)/Th(P) ~0.040) during the flood implies ongoing desorption in the coastal ocean, e.g. in the nearby oyster production areas (Marennes-Oléron Bay). The highest Ag(D) levels (6-8 ng/L) occurred in the mid-salinity range (15-20) of the Gironde Estuary and were decoupled from freshwater discharge. In the maximum turbidity zone, Ag(D) were at minimum, showing that high SPM concentrations (a) induce Ag(D) adsorption in estuarine freshwater and (b) counterbalance Ag(P) desorption in the low salinity range (1-3). Accordingly, Ag behaviour in turbid estuaries appears to be controlled by the balance between salinity and SPM levels. The first estimates of daily Ag(D) net fluxes for the Gironde Estuary (Boyle's method) showed relatively stable theoretical Ag(D) at zero salinity (Ag (D) (0) = 25-30 ng/L) for the contrasting hydrological situations. Accordingly, Ag(D) net fluxes were very similar for the situations with intermediate discharge (1.7 and 1.6 g/day) and clearly higher during the flood (5.0 g/day) despite incomplete desorption. Applying Ag (D) (0) to the annual freshwater inputs provided an annual net Ag(D) flux (0.64-0.89 t/year in 2008 and 0.56-0.77 t/year in 2009) that was 12-50 times greater than the Ag(D) gross flux. This estimate was consistent with net Ag(D) flux estimates obtained from gross Ag(P) flux considering 60 % desorption in the estuarine salinity gradient.

Synthesis, characterization and application of a new chelating resin for on-line separation, preconcentration and determination of Ag(I) by flame atomic absorption spectrometry.

A simple and reliable on-line separation/preconcentration procedure was developed for the determination of trace levels of Ag(I) by flame atomic absorption spectrometry. Poly[N-(3-methyl-1H-indol-1-yl)-2-methacrylamide-co-2-acrylamido-2-methyl-1-propane sulfonic acid-co-divinylbenzene] was synthesized and characterized as a new chelating resin for the first time. Ag(I) was sorbed on the chelating resin, from which it could be eluted with 3 mol L(-1) HCl and then introduced directly to the nebulizer-burner system for flame atomic absorption spectrometry. The parameters influential on the determination of Ag(I) ions such as the pH of the sample solution, amount of resin, eluent type, interfering ions and flow variables were studied. Under the optimum conditions, the calibration graph obtained was linear over the concentration range of 2-20 µg L(-1). The detection limit of the method (3σ) was 0.3 µg L(-1) while precision was 1.5% (n=25) at the level of 10 µg L(-1) Ag(I). The limit of quantification for the method, based on 20 σ, was 2.0 µg L(-1). The enrichment factor was found to be 65 while the optimized sample volume was 13.6 mL. The accuracy of the method was performed by analyzing certified reference materials. The developed method was applied successfully for the determination of silver in different water samples with satisfactory results.

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.