From bench to bedside: Improving the clinical safety of GalNAc-siRNA conjugates using seed-pairing destabilization.

Preclinical mechanistic studies have pointed towards RNA interference-mediated off-target effects as a major driver of hepatotoxicity for GalNAc-siRNA conjugates. Here, we demonstrate that a single glycol nucleic acid or 2'-5'-RNA modification can substantially reduce small interfering RNA (siRNA) seed-mediated binding to off-target transcripts while maintaining on-target activity. In siRNAs with established hepatotoxicity driven by off-target effects, these novel designs with seed-pairing destabilization, termed enhanced stabilization chemistry plus (ESC+), demonstrated a substantially improved therapeutic window in rats. In contrast, siRNAs thermally destabilized to a similar extent by the incorporation of multiple DNA nucleotides in the seed region showed little to no improvement in rat safety suggesting that factors in addition to global thermodynamics play a role in off-target mitigation. We utilized the ESC+ strategy to improve the safety of ALN-HBV, which exhibited dose-dependent, transient and asymptomatic alanine aminotransferase elevations in healthy volunteers. The redesigned ALN-HBV02 (VIR-2218) showed improved specificity with comparable on-target activity and the program was reintroduced into clinical development.

In Situ Synthesis of Horseradish Peroxidase Nanoflower@Carbon Nanotube Hybrid Nanobiocatalysts with Greatly Enhanced Catalytic Activity.

Organic-inorganic hybrid nanoflowers (NFs) consisting of horseradish peroxidase (HRP) and copper II (Cu2+) are successfully synthesized with the involvement of carbon nanotubes (CNTs) by in situ and post-modification methods. Catalytic activities of in situ synthesized HRP-NF@CNT (HRP-NF@CNT-Is) and post-modification-synthesized HRP-NF@CNTs (HRP-NF@CNT-Pm) are systematically examined. The 30 mg CNTs incorporated HRP-NF@CNT-Is (HRP-NF@CNT-30Is) exhibits greatly increased catalytic activity and stability toward 3,3',5,5'-tetramethylbenzidine (TMB), thanks to the synergistic effect between HRP-NF and CNTs and the peroxidase-like activity of CNTs in the presence of hydrogen peroxide (H2O2). While HRP-NF@CNT-30Is retains almost 85% of its initial activity even after 10 cycles, HRP-NF (without CNTs) loses half of its initial activity at the same experimental conditions. We study how two experimental parameters, the pH values and temperatures, influence the catalytic activity of HRP-NF@CNT-30Is, in addition to the fact that HRP-NF@CNT-30Is is employed to detect the presence of H2O2 and glutathione (GSH) with colorimetric and spectrophotometric readouts. For instance, HRP-NF@CNT-30Is is used to sensitively detect H2O2 in the range of 20 to 300 µM with an LOD of 2.26 µM. The catalytic activity of HRP-NF@CNT-30Is is suppressed in the presence of GSH, and then an obvious color change from blue to nearly colorless is observed. Using this strategy, GSH is also sensitively determined in the range of 20-200 µM with an LOD of 11.2 µM. We expect that HRP-NF@CNTs can be used as a promising and novel nanobiocatalyst for various biomedical and industrial applications in the near future.

Impacts of Chemical-Assisted Thermal Pretreatments on Methane Production from Fruit and Vegetable Harvesting Wastes: Process Optimization.

The increasing population creates excess pressure on the plantation and production of fruits and vegetables across the world. Consumption demand during the whole year has made production compulsory in the covered production system (greenhouse). Production, harvesting, processing, transporting, and distribution chains of fruit and vegetables have resulted in a huge amount of wastes as an alternative source to produce biofuels. In this study, optimization of two pretreatment processes (NaOH and HCl assisted thermal) was investigated to enhance methane production from fruit and vegetable harvesting wastes (FVHW) that originate from greenhouses. NaOH concentration (0-6.5%), HCl concentration (0-5%), reaction temperature (60-100 °C), solid content (1-5%), time of reaction (1-5 h), and mixing speed (0-500 rpm) were chosen in a wide range of levels to optimize the process in a broad design boundary and to evaluate the positive and negative impacts of independent variables along with their ranges. Increasing NaOH and HCl concentrations resulted in higher COD solubilization but decreased the concentration of soluble sugars that can be converted directly into methane. Thus, the increasing concentrations of NaOH and HCl in the pretreatments have resulted in low methane production. The most important independent variables impacting COD and sugar solubilization were found to be chemical concentration (as NaOH and HCl), solid content and reaction temperature for the optimization of pretreatment processes. The high amount of methane productions in the range of 222-365 mL CH4 gVS-1 was obtained by the simple thermal application without using chemical agents as NaOH or HCl. Maximum enhancement of methane production was 47-68% compared to raw FVHW when 5% solid content, 1-hour reaction time and 60-100 °C reaction temperature were applied in pretreatments.

Sequential coprecipitation and matrix removal for determination of cadmium impurities from multivitamin supplements by inductively coupled plasma mass spectrometry and method validation by isotope dilution analysis of SRM 3280 multivitamin/multielement tablets.

In this paper, we examined three different sequential coprecipitation schemes based on Mg(OH)2 and CaF2 precipitation using triethylamine (TEA) and hydrofluoric acid (HF), respectively, for determination of cadmium (Cd) impurities from multivitamin/mineral (MVM) supplements by isotope dilution (ID) inductively coupled plasma mass spectrometry (ICP-MS). The schemes involved three-step coprecipitation with either TEA alone or in combination with HF and are designated as Scheme 1 (TEA-TEA-TEA), Scheme 2 (TEA-HF-TEA) and Scheme 3 (HF-TEA-TEA) according to the addition sequence of each reagent. Experiments were carried out with MVM solutions spiked with 60 µg L-1 Cd from a multielement standard solution. All schemes provided quantitative separation of Cd from MVM matrix. Scheme 1 was the least effective in removal of interfering concomitant elements, molybdenum (Mo) and tin (Sn). Scheme 2 performed better for Sn, but failed in eliminating Mo. Scheme 3 was the most effective in eliminating both Mo and Sn. Mo levels in test MVM solutions reduced from 4.3 µg mL-1 to as low as 0.014 µg mL-1 while that for Sn decreased from 0.5 µg mL-1 to 0.018 µg mL-1 allowing interference-free determination of Cd to be achieved. Salt-matrix due to Mg, Ca, P and K along with the essential elements (Mn, Fe, Cu and Zn) levels was also reduced significantly. Reagent blanks from HF and TEA were insignificant (0.008 µg L-1) allowing a limit of detection of 0.004 µg L-1 or 0.26 ng g-1 Cd to be achieved (3σ, n = 6). The performance of the coprecipitation method (Scheme 3) was validated by determination of Cd in multivitamin/multielement tablets certified reference material (SRM 3280) by ID-ICP-MS. Experimental results (ng g-1) and recoveries were 78.8 ± 4.7 (98.5%), 77.9 ± 5.2 (97.4%) and 76.5 ± 4.8 (95.6%) for 110Cd, 111Cd and 114Cd isotopes, respectively. Several commercial MVM supplements were analyzed using the method. Mean Cd concentration ranged from 21.4 ng g-1 to 93.3 ng g-1. These values are much lower than those reported to date for various MVM supplements by ICP-MS determinations without chemical separation.

Efficient generation of volatile cadmium species using Ti(III) and Ti(IV) and application to determination of cadmium by cold vapor generation inductively coupled plasma mass spectrometry (CVG-ICP-MS).

In this study, a highly efficient chemical vapor generation (CVG) approach is reported for determination of cadmium (Cd). Titanium (III) and titanium (IV) were investigated for the first time as catalytic additives along with thiourea, L-cysteine and potassium cyanide (KCN) for generation of volatile Cd species. Both Ti(III) and Ti(IV) provided the highest enhancement with KCN. The improvement with thiourea was marginal (ca. 2-fold), while L-cysteine enhanced signal slightly only with Ti(III) in H2SO4. Optimum CVG conditions were 4% (v/v) HCl + 0.03 M Ti(III) + 0.16 M KCN and 2% (v/v) HNO3 + 0.03 M Ti(IV) + 0.16 M KCN with a 3% (m/v) NaBH4 solution. The sensitivity was improved about 40-fold with Ti(III) and 35-fold with Ti(IV). A limit of detection (LOD) of 3.2 ng L-1 was achieved with Ti(III) by CVG-ICP-MS. The LOD with Ti(IV) was 6.4 ng L-1 which was limited by the blank signals in Ti(IV) solution. Experimental evidence indicated that Ti(III) and Ti(IV) enhanced Cd vapor generation catalytically; for best efficiency mixing prior to reaction with NaBH4 was critical. The method was highly robust against the effects of transition metal ions. No significant suppression was observed in the presence of Co(II), Cr(III), Cu(II), Fe(III), Mn(II), Ni(II) and Zn(II) up to 1.0 µg mL-1. Among the hydride forming elements, no interference was observed from As(III) and Se(IV) at 0.5 µg mL-1 level. The depressive effects from Pb(II) and Sb(III) were not significant at 0.1 µg mL-1 while those from Bi(III) and Sn(II) were marginal. The procedures were validated with determination of Cd by CVG-ICP-MS in a number certified reference materials, including Nearshore seawater (CASS-4), Bone ash (SRM 1400), Dogfish liver (DOLT-4), Mussel tissue (SRM 2976) and Domestic Sludge (SRM 2781).

Is aceticlastic methanogen composition in full-scale anaerobic processes related to acetate utilization capacity?

In this study, biomass samples were obtained from six municipal and nine industrial full-scale anaerobic processes to investigate whether the aceticlastic methanogen population composition is related to acetate utilization capacity and the nature of the wastewater treated, i.e. municipal sludge or industrial wastewater. Batch serum bottle tests were used to determine the specific acetate utilization rate (AUR), and a quantitative real-time polymerase chain reaction protocol was used to enumerate the acetate-utilizing Methanosaeta and Methanosarcina populations in the biomass samples. Methanosaeta was the dominant aceticlastic methanogen in all samples, except for one industrial wastewater-treating anaerobic process. However, Methanosarcina density in industrial biomass samples was higher than the Methanosarcina density in the municipal samples. The average AUR values of municipal and industrial wastewater treatment plant biomass samples were 10.49 and 10.65 mg CH3COO(-)/log(aceticlastic methanogen gene copy).d, respectively. One-way ANOVA test and principle component analysis showed that the acetate utilization capacities and aceticlastic methanogen community composition did not show statistically significant correlation among the municipal digesters and industrial wastewater-treating processes investigated.

Selective solid phase extraction of copper using a new Cu(II)-imprinted polymer and determination by inductively coupled plasma optical emission spectroscopy (ICP-OES).

This work reports the preparation of a novel Cu(II)-ion imprinted polymer using 2-thiozylmethacrylamide (TMA) for on-line preconcentration of Cu(II) prior to its determination by inductively coupled optical emission spectroscopy (ICP-OES). Cu(II)-TMA monomer (complex) was synthesized and copolymerized via bulk polymerization method in the presence of ethyleneglycoldimethacrylate cross-linker. The resulting polymer was washed with 5% (v/v) HNO3 to remove Cu(II) ions and then with water until a neutral pH. The ion imprinted polymer was characterized by FT-IR and scanning electron microscopy. The experimental conditions were optimized for on-line preconcentration of Cu(II) using a minicolumn of ion imprinted polymer (IIP). Quantitative retention was achieved between pH 5.0 and 6.0, whereas the recoveries for the non-imprinted polymer (NIP) were about 61%. The IIP showed about 30 times higher selectivity to Cu(II) in comparison to NIP. The IIP also exhibited excellent selectivity for Cu(II) against the competing transition and heavy metal ions, including Cd, Co, Cr, Fe, Mn, Ni, Pb and Zn. Computational calculations revealed that the selectivity of IIP was mediated by the stability of Cu(II)-TMA complex which was far more stable than those of Co(II), Ni(II) and Zn(II) that have similar charge and ionic radii to Cu(II). A volume of 10 mL sample solution was loaded onto the column at 4.0 mL min-1 by using a sequential injection system (FIALab 3200) followed by elution with 1.0 mL of 2% (v/v) HNO3. The relative standard deviation (RSD) and limit of detection (LOD, 3s) of the method were 3.2% and 0.4 µg L-1, respectively. The method was successfully applied to determination of Cu(II) in fish otoliths (CRM 22), bone ash (SRM 1400) and coastal seawater and estuarine water samples.

Solid phase extraction of rare earth elements in seawater and estuarine water with 4-(2-thiazolylazo) resorcinol immobilized Chromosorb 106 for determination by inductively coupled plasma mass spectrometry.

A solid phase preconcentration method has been developed using new chelating resin prepared by immobilization of 4-(2-thiazolylazo) resorcinol (TAR) on Chromosorb 106. The method was optimized for determination of rare earth elements (REEs) in seawater and estuarine water samples by inductively coupled plasma mass spectrometry (ICP-MS). The effects of various experimental parameters, such as load pH, eluent concentration, sample and eluent flow rates were examined to find the optimum operating conditions. The REEs were quantitatively retained from saline solutions on a minicolumn Chromosorb 106-TAR resin at pH 5.0 and then eluted with 1.0 mL of 1% (v/v) HNO3. The resin possesses large sorption capacity for REEs ranging from 81.1 µmol g-1 for Lu and 108 µmol g-1 for Nd. Detection limits (3s) varied between 0.06 ng L-1 for Pr to 0.31 for Ce for preconcentration of 5.0 mL blank solutions (pH 5.0). The relative standard deviation for triplicate measurements was less than 5% at 0.1 µg L-1 level. The method was validated by analysis Nearshore seawater certified reference material (CASS-4). The elemental results were comparable with the values reported in literature. The method was verified by analysis of spiked and unspiked coastal seawater and estuarine water samples.

Effect of pretreated and anaerobically digested microalgae on the chemical and biochemical properties of soil and wheat grown on fluvisol.

ABSTRACTIn this study, the effects of the potential application of digestate as an agricultural fertiliser obtained from anaerobically digested microalgae treated by three pretreatment methods, namely alkaline hydrogen peroxide (AHP), high temperature and pressure (HTP), and hydrodynamic cavitation (HC) on some properties of soil, and wheat growth and yield were investigated. For this purpose, pretreated and anaerobically digested microalgae digestates alone or together with diammonium phosphate (DAP) as a chemical fertiliser were applied to soil for wheat growth. The highest dosage of AHP pretreated digestate combined with a half dose of DAP applied to soil was rich in nutrients as 0.25%N and 7.19 mg kg-1 compared to all groups. The properties of the soils were enhanced by applying the highest dosage (0.06 g kg-1) of microalgae digestate combined with a half dose of DAP. 0.02 g kg-1 dosage of HC pretreated digestate combined with a half dose of DAP also greatly improved nitrogen use efficiency indices by up to 104%. The soils' enzyme activities increased in wheat growth experiments by applying either raw or pretreated microalgae digestates. The soils' ß-glycosidase, alkaline phosphatase, and urease enzyme activities increased to 1.38 mg pNP g-1 soil, 4.91 mg pNP g-1 soil, and 2.27 mg NH4-N 100 g-1 soil respectively by the application of highest dosage of HC pretreated digestate. The digestates did not have a toxic effect on wheat growth, it was determined that applied pretreatment processes did not cause significant changes in wheat plant height or wet and dry weight.

On-line chemical vapour generation of cadmium in the presence of hexacyanochromate(III) for determination by inductively coupled plasma mass spectrometry (ICP-MS).

A vapour generation (VG) procedure has been described for determination of Cd by ICP-MS. Volatile species of Cd were generated on-line by interacting acidic sample solution containing potassium hexacyanochromate(III), K3Cr(CN)6, with sodium borohydride (NaBH4). The hexacyanochromate(III) complex was generated on-line by reacting 0.04 mol L-1 chromium(III) nitrate and 0.16 mol L-1 potassium cyanide (KCN) solutions in water. The resulting suspension of chromium(III) hydroxide, Cr(OH)3, was fed continuously to acidic stream of sample solution in the presence of excess KCN. The experimental conditions were optimized for effective generation of volatile species of Cd. Optimum signals were obtained from reaction of sample solutions in 4% v/v HCl with 2% m/v NaBH4 solution. Presence of K3Cr(CN)6 improved the efficiency of Cd vapour generation substantially affording 15-fold higher sensitivity. This phenomenon was thought to occur through formation of reactive intermediates evolved from interaction of [Cr(CN)6]3- with NaBH4 that react with Cd(II) to increase the yield volatile Cd species. Under the optimum conditions, no significant interferences were observed from the transition metals, including Cu and Ni, up to 1.0 µg mL-1 levels. Among the hydride forming elements, Bi, Pb, Sb and Sn depressed the signals above 0.1 µg mL-1. The detection limits (3s) were 6.2 and 5.2 ng L-1 for 110Cd and 111Cd isotopes, respectively. The method was successfully applied to determination of Cd by ICP-MS in several certified reference materials, including Nearshore seawater (CASS-4), Bone ash (SRM 1400), Dogfish liver (DOLT-4) and Mussel tissue (SRM 2976).

Solid phase extraction of Cu(II), Ni(II), Pb(II), Cd(II) and Mn(II) ions with 1-(2-thiazolylazo)-2-naphthol loaded Amberlite XAD-1180.

A new method for separation and preconcentration of trace amounts of Cu(II), Ni(II), Pb(II), Cd(II) and Mn(II) ions in various matrices was proposed. The method is based on the adsorption and chelation of the metal ions on a column containing Amberlite XAD-1180 resin impregnated with 1-(2-thiazolylazo)-2-naphthol (TAN) reagent prior to their determination by flame atomic absorption spectrometry (FAAS). The effect of pH, type, concentration and volume of eluent, sample volume, flow rates of sample and elution solutions, and interfering ions have been investigated. The optimum pH for simultaneous retention of all the metal ions was 9. Eluent for quantitative elution was 20 ml of 2 mol l(-1) HNO(3). The optimum sample and eluent flow rates were found as 4 ml min(-1), and also sample volume was 500 ml, except for Mn (87% recovery). The sorption capacity of the resin was found to be 0.77, 0.41, 0.57, and 0.30 mg g(-1) for Cu(II), Ni(II), Cd(II), and Mn(II), respectively. The preconcentration factor of the method was 200 for Cu(II), 150 for Pb(II), 100 for Cd(II) and Ni(II), and 50 for Mn(II). The recovery values for all of the metal ions were > or = 95% and relative standard deviations (RSDs) were < or = 5.1%. The detection limit values were in the range of 0.03 and 1.19 microg l(-1). The accuracy of the method was confirmed by analysing the certified reference materials (TMDA 54.4 fortified lake water and GBW 07605 tea samples) and the recovery studies. This procedure was applied to the determination of Cu(II), Ni(II), Pb(II), Cd(II) and Mn(II) in waste water and lake water samples.

Synthesis of Long-Term Stable Gold Nanoparticles Benefiting from Red Raspberry (Rubus idaeus), Strawberry (Fragaria ananassa), and Blackberry (Rubus fruticosus) Extracts-Gold Ion Complexation and Investigation of Reaction Conditions.

We report synthesis of monodispersed, stable, and colloidal gold nanoparticles (Au NPs) using anthocyanin-riched red raspberry (Rubus idaeus), strawberry (Fragaria ananassa), and blackberry (Rubus fruticosus) extracts as functions of concentration of HAuCl4·3H2O and berries extract, reaction time, and reaction pH values (pHs) and demonstrate their unique stability in highly concentrated salt (sodium chloride, NaCl) solutions. The catecholamine group of anthocyanin molecules give preferential coordination reaction with gold ions (Au3+) for creating anthocyanin-Au3+ complexes, which may lead to initiation of nucleation for seed formation, and then, oxidation of catecholamine results in a flow of electrons from anthocyanins to Au seeds for anisotropic growth. Finally, the surface of the Au NPs is saturated with anthocyanins, and formation of monodispersed and stable Au NPs with narrow size distribution is completed. We also report the effects of some experimental parameters including concentrations of Au3+ ions and barrier extracts, reaction time, and pHs on formation of the Au NPs with rational explanations. The long-term colloidal stability of the Au NPs in the 400 mM NaCl solution was comparatively studied with commercial Au NPs (citrate capped). As results show that anthocyanin-riched berry extracts directed Au NPs we proposed here can be considered as promising and safe tools for biomedical applications owing to their highly much colloidal dispersibility and salt tolerance properties.

Selection of GalNAc-conjugated siRNAs with limited off-target-driven rat hepatotoxicity.

Small interfering RNAs (siRNAs) conjugated to a trivalent N-acetylgalactosamine (GalNAc) ligand are being evaluated in investigational clinical studies for a variety of indications. The typical development candidate selection process includes evaluation of the most active compounds for toxicity in rats at pharmacologically exaggerated doses. The subset of GalNAc-siRNAs that show rat hepatotoxicity is not advanced to clinical development. Potential mechanisms of hepatotoxicity can be associated with the intracellular accumulation of oligonucleotides and their metabolites, RNA interference (RNAi)-mediated hybridization-based off-target effects, and/or perturbation of endogenous RNAi pathways. Here we show that rodent hepatotoxicity observed at supratherapeutic exposures can be largely attributed to RNAi-mediated off-target effects, but not chemical modifications or the perturbation of RNAi pathways. Furthermore, these off-target effects can be mitigated by modulating seed-pairing using a thermally destabilizing chemical modification, which significantly improves the safety profile of a GalNAc-siRNA in rat and may minimize the occurrence of hepatotoxic siRNAs across species.

Novel Imprinted Polymer for the Preconcentration of Cadmium with Determination by Inductively Coupled Plasma Mass Spectrometry.

A novel Cd(II)-imprinted polymer was prepared with chemical immobilization approach by using N-methacryloyl-L-histidine as a vinylated chelating agent for on-line solid phase extraction of Cd(II) for determination by inductively coupled plasma mass spectrometry. Cd(II)-monomer complex was synthesized and copolymerized via bulk polymerization method in the presence of ethyleneglycoldimethacrylate cross-linker. The resulting polymer was leached with 1.0 mol L-1 HNO3 to generate the cavities in the polymer for Cd(II) ions. The experimental conditions, including load pH, solution flow rate, and eluent concentration for effective sorption of Cd(II) were optimized using a minicolumn of the imprinted polymer. A volume of 5.0 mL sample 5 µg L-1 Cd(II) solution at pH 6.5 was loaded onto the column at 2.0 mL min-1 by using a sequential injection system (FIALab 3200) followed by elution with 1.0 mL of 0.75 mol L-1 HNO3. The relative selectivity coefficients of the imprinted polymer for Cd(II) were 38.5, 3.5, 3.0, 2.5 and 6.0 in the presence of Cu(II), Ni(II), Zn(II), Co(II) and Pb(II), respectively. Computational calculations revealed that the selectivity of the imprinted polymer was mediated by the stability of Cd(II)-N-methacryloyl-L-histidine complex which was far more stable than those of commonly used monomers, such as 4-vinyl pyridine, methacrylic acid and vinylimidazole. The detection limit (3s) and relative standard deviation (%) were found to be 0.004 µg L-1 and 3.2%, respectively. The method was validated by analysis of seawater certified reference material (CASS-4) and successfully applied to the determination of Cd(II) in coastal seawater and estuarine water samples.

Self assembled snowball-like hybrid nanostructures comprising Viburnum opulus L. extract and metal ions for antimicrobial and catalytic applications.

Herein, we report the production of novel organic-inorganic nanobio-catalytic and antimicrobial agents called "nanosnowball" (NSBs) with a rational design and elucidate the increase in the catalytic and antimicrobial activities. The NSBs resemble to guelder rose were formed of the extract of Viburnum opulus (VO) as the organic component and copper (II) ions (Cu2+) as the inorganic component. The effects of the VO extract and Cu2+ concentrations on the morphology of NSBs were systematically examined and characterized with several techniques such as SEM, FT-IR, EDX and XRD. Our results demonstrated that the presence of CuO bonds in NSBs could be indication of VO extract-Cu2+ complexes. Interestingly, the NSBs exhibited peroxidase-like activity towards guaiacol used as a model substrate depending on Fenton-like reaction. While free VO extract did not show antimicrobial activity at indicated concentration (2000-125µg/mL), the NSBs showed effective antimicrobial activity against bacterial (Escherichia coli ATCC 35218, Salmonella typhi ATCC 14028, Enterococcus faecium ATCC 8459, Enterococcus faecalis ATCC 29212, Bacillus cereus ATCC 11778, Staphylococcus aureus ATCC 25923, except Pseudomonas aeruginosa ATCC 27853 and Haemophilus influenza ATCC 49247) and fungal pathogens (Candida albicans ATCC 10231, C. glabrata ATCC 90030), respectively.

DNA aptamer functionalized gold nanostructures for molecular recognition and photothermal inactivation of methicillin-Resistant Staphylococcus aureus.

In this work, we report the development of DNA aptamer-functionalized gold nanoparticles (Apt@Au NPs) and gold nanorods (Apt@Au NRs) for inactivation of Methicillin-resistant Staphylococcus aureus (MRSA) with targeted photothermal therapy (PTT). Although both Apt@Au NPs and Apt@Au NRs specifically bind to MRSA cells, Apt@Au NPs and Apt@Au NRs inactivated ∼5% and over 95% of the cells,respectively through PTT. This difference in inactivation was based on the relatively high longitudinal absorption of near-infrared (NIR) radiation and strong photothermal conversion capability for the Apt@Au NRs compared to the Apt@Au NPs. The Au NRs served as a nanoplatform for the loading of thiolated aptamer and also provided multivalent effects for increasing binding strength and affinity to MRSA. Our results indicate that the type of aptamer and the degree of multivalent effect(s) are important factors for MRSA inactivation efficiency in PTT. We show that the Apt@Au NRs are a very effective and promising nanosystem for specific cell recognition and in vitro PTT.

MULTIELEMENT SOLID PHASE PRECONCENTRATION USING A CHELATING RESIN OF STYRENE DIVINYLBENZENE COPOLYMER AND APPLICATION TO ANALYSIS OF SEAWATER AND FISH OTOLITHS BY INDUCTIVELY COUPLED PLASMA MASS SPECTROMETRY (ICP�MS).

A new chelating resin has been synthesized by immobilizing 4-(2-thiazolylazo) resorcinol (TAR) onto styrene divinlybenzene copolymer and examined for on-line solid phase extraction/preconcentration of Cd, Co, Cu, Ni, Pb and Zn in seawater and fish otoliths for determination by inductively plasma mass spectrometry (ICP-MS). A volume of 5.0 mL sample solution was loaded onto the mini column of TAR immobilized resin at 2.0 mL min-1 via a sequential injection system. The optimum pH for multielement preconcentration was around pH 5.5. Recoveries were better than 96% in artificial seawater. Elution was achieved with 1.0 mL of 0.75 mol L-1 HNO3. The resin possesses large sorption capacity ranging from 82.0 µmol g-1 for Pb to 319 µmol g-1 for Cu. The detection limits (3s) varied between 0.0016 µg L-1 (Cd) and to 0.015 µg L-1 (Zn) for preconcentration of 5.0 mL blank solutions (pH 5.5). Relative standard deviation (RSD)for three replicate runs was between 0.3% (Cd) and 6% (Zn) at 1.0 µg L-1 level. The procedure was validated by analysis of Nearshore Seawater certified reference material (CASS-4), and then successfully applied to the determination of the trace elements in fish otoliths (CRM 22) and in coastal seawater and estuarine water samples.

Determination of lead by hydride generation inductively coupled plasma mass spectrometry (HG-ICP-MS): on-line generation of plumbane using potassium hexacyanomanganate(III).

A hydride generation (HG) procedure has been described for determination of Pb by ICP-MS using potassium hexacyanomanganate(III), K(3)Mn(CN)(6), as an additive to facilitate the generation of plumbane (PbH(4)). Potassium hexacyanomanganate(III) was prepared in acidic medium as it was unstable in water. The stability of hexacyanomanganate(III) was examined in dilute solutions of HCl, HNO(3) and H(2)SO(4). The solutions prepared in 1% v/v H(2)SO(4) were found to be stable for over a period of 24h. The least suitable medium was 1% v/v HNO(3). For generation of plumbane, acidic hexacyanomanganate(III) and sample solutions were mixed on-line along a 5-cm long tygon tubing (1.14 mm i.d.) and then reacted with 2% m/v sodium borohydride (NaBH(4)). A concentration of 0.5% m/v K(3)Mn(CN)(6) facilitated the generation of PbH(4) remarkably. In comparison to H(2)SO(4), HCl provided broader working range for which optimum concentration was 1% v/v. No significant interferences were noted from transition metals and hydride forming elements, up to 0.5 µg mL(-1) levels, except Cu which depressed the signals severely. The depressive effects in the presence of 0.1 µg mL(-1) Cu were alleviated by increasing the concentration of K(3)Mn(CN)(6) to 2% m/v. Under these conditions, the sensitivity was enhanced by a factor of at least 42 to 48. The detection limit (3s) was 0.008 µg L(-1) for (208)Pb isotope. Average signal-to-noise ratio (S/N) ranged between 18 and 20 for 1.0 µg mL(-1) Pb solution. The accuracy of the method was verified by analysis of several certified reference materials, including Nearshore seawater (CASS-4), Bone ash (SRM 1400), and Mussel tissue (SRM 2976). The procedure was also successfully applied to the determination of Pb in coastal seawater samples by ICP-MS.

Synthesis, characterization and application of a novel ion-imprinted polymer for selective solid phase extraction of copper(II) ions from high salt matrices prior to its determination by FAAS.

A new Cu(II)-imprinted sorbent has been prepared by using 5-methyl-2-thiozylmethacrylamide (MTMAAm). The monomer of Cu(II)-MTMAAm complex was synthesized and copolymerized in the presence of ethyleneglycol dimethacrylate cross-linker via bulk polymerization method. The resulting Cu(II)-imprinted polymer was characterized by FT-IR spectroscopy and scanning electron microscopy (SEM). Copper ions were removed from the polymer with 1.0 mol L(-1) HNO3 and determined by flame atomic absorption spectrometry (FAAS). The imprinted polymer showed higher selectivity for Cu(II) in comparison to the non-imprinted polymer. Relative selectivity coefficients (k') for Cu(II)/Zn(II), Cu(II)/Ni(II) and Cu(II)/Co(II) were 9.1, 14.8 and 26.6, respectively. The imprinted polymer was examined as a column packing material for solid phase extraction of Cu(II) from various matrices. The effects of solution pH, acid eluents and interfering ions were investigated. The poylmer possesses selective extraction of Cu(II) within pH range from 5.0 to 6.5. The relative standard deviation and limit of detection (3s) of the method were evaluated as 1.4% and 0.9 µg L(-1), respectively. The accuracy of the method was verified by analysis of two certified reference materials (CWW-TM-D and SRM 3280) and then applied to the determination of Cu in seawater, lake water and tap water samples, and hemodialysis concentrates and multivitamin/multielement supplements.

Cyanovanadate(III) complexes as novel additives for efficient generation of volatile cadmium species in complex samples prior to determinations by inductively coupled plasma mass spectrometry (ICP-MS).

A new method has been described for generation of volatile species of Cd using vanadium(III) cyanide complex. Aqueous solutions of 0.04 mol L(-1) vanadium chloride (VCl3) and 0.12 mol L(-1) potassium cyanide (KCN) were reacted on-line yielding a suspension of vanadium hydroxide, V(OH)3. This suspension was dissolved along the stream of sample solution in dilute HCl to form heptacyanovanadate(III) complex, [V(CN)7]4-. Volatile Cd species were generated by reacting the stream of sample solution and cyanovanadate(III) complex with sodium borohydride (NaBH4). Feasibility of off-line and on-online approaches was investigated for quantitative determinations. Better precision and daily stability were achieved with on-line settings. Optimum signals were obtained from sample solutions within a range of 3 to 5% v/v HCl. A concentration of 2% m/v NaBH4 was adequate to achieve an enhancement of 20-fold in the presence of cyanovanadate(III) complex. The limits of detection were 5.0 and 4.5 ng L(-1) for 110Cd and 111Cd isotopes, respectively. Precision (%RSD) was better than 4.7% for six replicate measurements. The interferences of Cu(II) and Ni(II) were marginal (<10%) at 1.0 µg mL(-1). Depressive effects from Bi, Se and Sn were not significant below 0.1 µg mL(-1). The method was validated by determination of Cd using ICP-MS in certified reference materials of Nearshore seawater (CASS-4), Bone ash (SRM 1400), Dogfish liver (DOLT-4) and Mussel tissue (SRM 2976).