Analysis of perchlorate in human urine using ion chromatography and electrospray tandem mass spectrometry

Because of health concerns surrounding widespread exposure to perchlorate, we developed a sensitive and selective method for quantifying perchlorate in human urine using ion chromatography coupled with electrospray ionization tandem mass spectrometry. Perchlorate was quantified using a stable isotope-labeled internal standard ((18)O(4)-perchlorate) with excellent assay precision (coefficient of variation <5% for repetitively analyzed quality control material). Analytical accuracy was established by blind analysis of certified proficiency testing materials prepared in synthetic urine matrix; calculated amounts deviated minimally from true amounts, with percent differences ranging from 2% to 5%. Selective chromatography and tandem mass spectrometry reduced the need for sample cleanup, resulting in a rugged and rapid method capable of routinely analyzing 75 samples/day. The lowest reportable level (0.025 ng/mL) was sufficiently sensitive to detect perchlorate in all human urine samples evaluated to date, with a linear response range from 0.025 to 100 ng/mL. This selective, sensitive, and rapid method will help elucidate any potential associations between human exposure to low levels of perchlorate and adverse health effects.     

Mapping invasive Phragmites australis in the coastal Great Lakes with ALOS PALSAR satellite imagery for decision support

The invasive variety of Phragmites australis (common reed) forms dense stands that can cause negative impacts on coastal Great Lakes wetlands including habitat degradation and reduced biological diversity. Early treatment is key to controlling Phragmites, therefore a map of the current distribution is needed. ALOS PALSAR imagery was used to produce the first basin-wide distribution map showing the extent of large, dense invasive Phragmites-dominated habitats in wetlands and other coastal ecosystems along the U.S. shore of the Great Lakes. PALSAR is a satellite imaging radar sensor that is sensitive to differences in plant biomass and inundation patterns, allowing for the detection and delineation of these tall (up to 5 m), high density, high biomass invasive Phragmites stands. Classification was based on multi-season ALOS PALSAR L-band (23 cm wavelength) HH and HV polarization data. Seasonal (spring, summer, and fall) datasets were used to improve discrimination of Phragmites by taking advantage of phenological changes in vegetation and inundation patterns over the seasons. Extensive field collections of training and randomly selected validation data were conducted in 2010–2011 to aid in mapping and for accuracy assessments. Overall basin-wide map accuracy was 87%, with 86% producer's accuracy and 43% user's accuracy for invasive Phragmites. The invasive Phragmites maps are being used to identify major environmental drivers of this invader's distribution, to assess areas vulnerable to new invasion, and to provide information to regional stakeholders through a decision support tool.     

Disinfection of Bacillus subtilis spores with chlorine dioxide: a bench-scale and pilot-scale study

Chlorine dioxide (ClO2) inactivation of Bacillus subtilis ATCC 19659 spores was examined at pilot-scale during periods representative of winter and summer temperature extremes at the Britannia Water Treatment Plant in Ottawa, Canada. In addition, bench-scale experiments using the same source water (Ottawa River, Ontario, Canada), as well as buffered and unbuffered laboratory waters were conducted using B. subtilis spores. Bench-scale inactivation of B. subtilis spores by ClO2 was similar to reported values for Cryptosporidium parvum (both organisms being more resistant to ClO2 than Giardia lamblia), suggesting the possibility that these spores may be used as potential indicators for protozoan parasites. Additionally, spore inactivation was observed to be influenced by pH in laboratory (distilled deionised water) water but not in Ottawa River water. At pilot-scale, spore inactivation was influenced by water temperature: a ClO2 dose of 2.5 mg/L resulted in a spore inactivation of approximately 2.0 log10 and 0.5 log10 at water temperatures of 23.2d egrees C and 5.2 degrees C, respectively. Chlorite concentrations remained below both the US EPA maximum contaminant level of 1.0 mg/L and the maximum contaminant level goal of 0.8 mg/L for up to 2.0log10 B. subtilis inactivation.     

Optimization of Supercritical Carbon Dioxide Extraction of Bioactive Flavonoid Compounds from Spearmint (<Emphasis Type="Italic">Mentha spicata</Emphasis> L.) Leaves by Using Response Surface Methodology

The bioactive flavonoid compounds of spearmint (Mentha spicata L.) leaves were obtained by using supercritical carbon dioxide (SC-CO₂) extraction. Extraction was carried out according to face-centred central composite design, and independent variables were pressure (100, 200 and 300 bar), temperature (40, 50 and 60 °C) and co-solvent amount (3, 6 and 9 g/min). The extraction process was optimized by using response surface methodology for the highest crude extraction yield of bioactive flavonoid compounds. The optimal conditions were identified as 209.39 bar pressure, 50.00 °C temperature and 7.39 g/min co-solvent amount. The obtained extract under optimum SC-CO₂ condition was analysed by high-performance liquid chromatography. Seven bioactive flavonoids including catechin, epicatechin, rutin, luteolin, myricetin, apigenin and naringenin were identified as major compounds. The results of quantification showed that spearmint leaves are potential source of antioxidant compounds.     

Application of 2-hydroxypropyl-β-cyclodextrin in the assay of acyl-CoA: cholesterol acyltransferase and neutral and acid cholesterol ester hydrolases

The utility of 2-hydroxypropyl-β-cyclodextrin for increasing the sensitivity of assays for the microsomal acyl-CoA:cholesterol acyltransferase, and the acid lysosomal and the neutral microsomal and cytosolic cholesterol ester hydrolase activity was studied in rat hepatocytes. Enzyme assays, at optimal concentrations of cyclodextrin, were validated by assessing: (i) linearity of product formation with incubation time and protein amount, and saturation with substrate, and (ii) the effect of treatments of cells or of subcellular fractions on enzyme activities. Delivery of cholesterol dissolved in 2-hydroxypropyl-β-cyclodextrin to the acyl-CoA:cholesterol acyltransferase assay mixture raised the enzyme activity more than 8-fold and was twice that measured when cholesterol was added in Triton WR-1339. 2-Hydroxypropyl-β-cyclodextrin itself was partially effective, apparently by making endogenous cholesterol more accesible to the enzyme. Inclusion of 2-hydroxypropyl-β-cyclodextrin in cholesterol ester hydrolase assays using standard micellar substrates doubled the activity estimated in lysosome and microsome preparations and enhanced the cytosolic cholesterol esterase activity by about 50%. Differences in the catalytic activity of acyl-CoA:cholesterol acyltransferase and cholesterol ester hydrolases caused by treatment of hepatocytes with compound 58-035 or 25-hydroxycholesterol, or of subcellular fractions with NaF, were maintained when enzymes were assayed with cyclodextrin. The results indicate that 2-hydroxypropyl-β-cyclodextrin is a suitable vehicle for delivering cholesterol to acyl-CoA:cholesterol acyltransferase and enhances the sensitivity of standard assays of the enzymes governing the intrahepatic hydrolysis of cholesteryl esters.     

To Extinguish the Fire from Outside the Cell or to Shutdown the Gas Valve Inside? Novel Trends in Anti-Inflammatory Therapies

Cytokines are the most important soluble mediators of inflammation. Rare pediatric diseases provided exemplar conditions to study the anti-inflammatory efficacy of new generation therapies (biologics/biopharmaceuticals) selectively targeting single cytokines. Monoclonal antibodies and recombinant proteins have revolutionized anti-inflammatory therapies in the last two decades, allowing the specific targeting of single cytokines. They are very effective in extinguishing inflammation from outside the cell, even with the risk of an excessive and prolonged immunosuppression. Small molecules can enter the cell and shutdown the valve of inflammation by directly targeting signal proteins involved in cytokine release or in response to cytokines. They are orally-administrable drugs whose dosage can be easily adjusted to obtain the desired anti-inflammatory effect. This could make these drugs more suitable for a wide range of diseases as stroke, gout, or neurological impairment, where inflammatory activation plays a pivotal role as trigger. Autoinflammatory diseases, which have previously put anti-cytokine proteins in the limelight, can again provide a valuable model to measure the real potential of small inhibitors as anti-inflammatory agents.     

HATMSC Secreted Factors in the Hydrogel as a Potential Treatment for Chronic Wounds—In Vitro Study

Mesenchymal stem cells (MSCs) can improve chronic wound healing; however, recent studies suggest that the therapeutic effect of MSCs is mediated mainly through the growth factors and cytokines secreted by these cells, referred to as the MSC secretome. To overcome difficulties related to the translation of cell therapy into clinical use such as efficacy, safety and cost, we propose a hydrogel loaded with a secretome from the recently established human adipose tissue mesenchymal stem cell line (HATMSC2) as a potential treatment for chronic wounds. Biocompatibility and biological activity of hydrogel-released HATMSC2 supernatant were investigated in vitro by assessing the proliferation and metabolic activity of human fibroblast, endothelial cells and keratinocytes. Hydrogel degradation was measured using hydroxyproline assay while protein released from the hydrogel was assessed by interleukin-8 (IL-8) and macrophage chemoattractant protein-1 (MCP-1) ELISAs. Pro-angiogenic activity of the developed treatment was assessed by tube formation assay while the presence of pro-angiogenic miRNAs in the HATMSC2 supernatant was investigated using real-time RT-PCR. The results demonstrated that the therapeutic effect of the HATMSC2-produced factors is maintained following incorporation into collagen hydrogel as confirmed by increased proliferation of skin-origin cells and improved angiogenic properties of endothelial cells. In addition, HATMSC2 supernatant revealed antimicrobial activity, and which therefore, in combination with the hydrogel has a potential to be used as advanced wound-healing dressing.     

Tuning the optical bandgap of multi-walled carbon nanotube-modified zinc silicate glass-ceramic composites

Novel glass-ceramic composites with optical bandgap tunability were synthesised. Zinc silicate powder (ZS) was mixed with multi-walled carbon nanotubes (MWCNTs) at various mass fractions (0, 1, 2, and 3 wt %), followed by argon sintering. X-ray diffraction (XRD) analysis revealed the structural change from an amorphous ZS phase to a crystalline willemite phase (Zn₂SiO₄) by adding MWCNTs, and the largest crystallite size was obtained for ZS with 2.0 wt% MWCNTs. Although the agglomeration of ZS and MWCNTs was observed by field emission scanning electron microscopy (FESEM), there was no chemical interaction between ZS and MWCNTs as confirmed by Fourier transform infrared spectroscopy (FTIR). MWCNTs enhanced the crystallisation, which led to the green emission of Zn₂SiO₄ blue-shifting from 572 nm to 557 nm. The narrowed optical bandgap of Zn₂SiO₄ was attributed to the MWCNT-induced exciton localised between the valence band and conduction band of Zn₂SiO₄. The bandgap tuning effect of MWCNTs potentially paved new ways to mass fabricate zinc silicate-based semiconductors with desirable optical bandgap energy E_g, which significantly benefits the sensor and laser-related industry.     

Synthesis of Edible Vegetable Oils Enriched with Healthy 1,3-Diglycerides Using Crude Glycerol and Homogeneous/Heterogeneous Catalysis

Commercial edible vegetable oils in which part of their triglycerides are substituted with 1,3-diglycerides are healthier for human consumption than the original oils. This is because the human metabolism of 1,3-diglycerides is believed to occur through a distinct pathway with less probability of being deposited as fat in the body tissues. To obtain these enriched oils, conversion of triglycerides into diglycerides is carried out by glycerolysis using commercial crude glycerol containing dissolved alkali cations that homogeneously catalyze the reaction. The addition of a food production-compatible MgO as a supplementary solid basic catalyst, shortens the reaction time by half due to a combination of homogeneous and heterogeneous catalysis processes. In either homogeneously or homogeneous-heterogeneously catalyzed glycerolysis, the increase of the reaction temperature in the range of 453–493 K increases the final 1,3-diglyceride content. Furthermore, in both glycerolysis processes the triglyceride content can be decreased in more than 60% with the consequent increase of total diglycerides to 50%, 70% of which are the 1,3-isomers. The glycerolysis reaction proceeds without altering the fatty acid distribution of the original oils.