Determination of methoxyphenols in ambient atmospheric particulate matter: tracers for wood combustion

Combustion of wood and other biomass fuels produces source-specific organic compounds arising from pyrolysis of lignin, including substantial amounts of 4-substituted methoxylated phenolic compounds (methoxyphenols). These compounds have been used as atmospheric markers to determine the contribution of wood smoke to ambient atmospheric fine particulate matter (PM). However, reliable quantification of methoxyphenols represents an analytical challenge because these compounds are polar, semi-volatile, and somewhat reactive. We reportherein an improved gas chromatographic-mass spectrometric (GC/MS) method for the sensitive and reliable determination of methoxyphenols in low-volume ambient PM samples. Deuterated standard compounds are added to the environmental samples prior to extraction to determine analyte recoveries in each sample. Analytical figures of merit for the assay, as applied to ambient PM2.5 and PM10 samples are as follows: recovery = 63-100%; precision = 2-6%; analytical limit of detection (S/N 2) = 0.002 microg/mL; limit of quantitation = 0.07-0.45 ng/m3 (assuming a 14 m3 sample). The improved method was applied to ambient PM samples collected between 1999 and 2000 in Seattle, WA. Particle-bound methoxyphenol concentrations in the range <0.1 to 22 ng/m3 were observed and the methoxyphenols were present almost exclusively in the fine (PM2.5) size fraction. We also demonstrated that XRF analysis of samples of atmospheric PM collected on Teflon filters significantly reduced the levels of methoxyphenols measured in the PM samples in subsequent assay of the same filters. Therefore, XRF analysis of filters, commonly undertaken to obtain trace element concentrations for use in source apportionment analyses, would preclude the subsequent analysis of those filters for methoxyphenols and other similarly semivolatile or reactive organic chemicals.     

Biocatalytic oxidase: Batch to continuous

The adoption of more efficient development strategies and manufacturing techniques will be essential for future success in the bio manufacturing sectors. Continuous operation of biocatalytic processes has the potential to offer many advantages over established batch process methodologies. There exist opportunities for improved process control; ease of scale up; minimizing of interruptions in production; reducing reactor size; and economic use of biocatalysts.     

Concentration and preservation of very low abundance biomarkers in urine, such as human growth hormone (hGH), by Cibacron Blue F3G-A loaded hydrogel particles

Urine is a potential source of diagnostic biomarkers for detection of diseases, and is a very attractive means of non-invasive biospecimen collection. Nonetheless, proteomic measurement in urine is very challenging because diagnostic biomarkers exist in very low concentration (usually below the sensitivity of common immunoassays) and may be subject to rapid degradation. Hydrogel nanoparticles functionalized with Cibacron Blue F3G-A (CB) have been applied to address these challenges for urine biomarker measurement. We chose one of the most difficult low abundance, but medically relevant, hormones in the urine: human growth hormone (hGH). The normal range of hGH in serum is 1 to 10 ng/mL but the urine concentration is suspected to be a thousand times less, well below the detection limit (50 pg/mL) of sensitive clinical hGH immunoassays. We demonstrate that CB particles can capture, preserve and concentrate hGH in urine at physiological salt and urea concentrations, so that hGH can be measured in the linear range of a clinical immunometric assay. Recombinant and cadaveric hGH were captured from synthetic and human urine, concentrated and measured with an Immulite chemiluminescent immunoassay. Values of hGH less than 0.05 ng/mL (the Immulite detection limit) were concentrated to 2 ng/mL, with a urine volume of 1 mL. Dose response studies using 10 mL of urine demonstrated that the concentration of hGH in the particle eluate was linearly dependent on the concentration of hGH in the starting solution, and that all hGH was removed from solution. Thus if the starting urine volume is 100 mL, the detection limit will be 0.1 pg/mL. Urine from a healthy donor whose serum hGH concentration was 1.34 ng/mL was studied in order detect endogenous hGH. Starting from a volume of 33 mL, the particle eluate had an hGH concentration of 58 pg/mL, giving an estimated initial concentration of hGH in urine of 0.175 pg/mL. The nanotechnology described here appears to have the desired precision, accuracy and sensitivity to support large scale clinical studies of urine hGH levels.      

Real-time tracking of CO₂ injected into a subsurface coal fire through high-frequency measurements of the ¹³CO₂ signature

CO? was injected into a coal fire burning at a depth of 15 m in the subsurface in southwestern Colorado, USA. Measurements were made of the 13CO? isotopic signature of gas exhaust from an observation well and two surface fissures. The goal of the test was to determine (1) whether CO? with a distinct isotopic signature could be used as a tracer to identify flow pathways and travel times in a combustion setting where CO? was present in significant quantities in the gases being emitted from the coalbed fire, and (2) to confirm the existence of a self-propagating system of air-intake and combustion gas exhaust that has been previously proposed. CO? was injected in three separate periods. The 13CO? isotopic signature was measured at high frequency (0.5 Hz) before, during, and after the injection periods for gas flowing from fissures over the fire and from gas entering an observation well drilled into the formation just above the fire but near the combustion zone. In two cases, a shift in the isotopic signature of outgassing CO? provided clear evidence that injected CO? had traveled from the injection well to the observation point, while in a third case, no response was seen and the fissure could not be assumed to have a flowpath connected with the injection well. High-frequency measurements of the 13CO? signature of gas in observation wells is identified as a viable technique for tracking CO? injected into subsurface formations in real-time. In addition, a chimney-like coupled air-intake and exhaust outlet system feeding the combustion of the coal seam was confirmed. This can be used to further develop strategies for extinguishing the fire.     

Attachment of Listeria monocytogenes on ready-to-eat meats

Five individual strains of Listeria monocytogenes and a mixed cocktail of all five were studied for attachment on frankfurters, ham, bologna, and roast beef relative to their cell surface characteristics. The ratio of strongly attached (sessile) L. monocytogenes cells compared with total (sessile and planktonic) attached cells on ready-to-eat meats was also determined. Because bacterial cell surfaces were characterized by net negative charge and hydrophobicity, electrostatic interaction chromatography and cationized ferritin methods were chosen to study net negative charge distribution on the bacterial cell surface, whereas hydrophobic interaction chromatography and contact angle measurement were used to examine the cell surface hydrophobicity. No differences (P > 0.05) were observed in cell surface charge or cell surface hydrophobicity among strains. Approximately 84 to 87% L. monocytogenes were found to attach strongly to ready-to-eat meats within 5 min. No differences (P > 0.05) were found among strains or among meats. Micrographs observed from scanning electron microscopy showed no differences among the strains but showed a difference in age of cells (mixed culture) in terms of surface negative charge distribution. More surface negatively charged sites were observed at 0 and 7 days and much fewer at 3 days during storage of washed, harvested cells in buffer at 4 degrees C (aged cells under cold and nutrient deprivation), indicating a possible change in cell surface properties. Because no difference in strains was observed, the contact angle measurement study was carried out with the five-strain mixed culture. The surface hydrophobicity increased in frankfurters, decreased in roast beef, and was unchanged in ham and bologna as a result of inoculation.     

Source apportionment of PM(2.5) and selected hazardous air pollutants in Seattle

The potential benefits of combining the speciated PM(2.5) and VOCs data in source apportionment analysis for identification of additional sources remain unclear. We analyzed the speciated PM(2.5) and VOCs data collected at the Beacon Hill in Seattle, WA between 2000 and 2004 with the Multilinear Engine (ME-2) to quantify source contributions to the mixture of hazardous air pollutants (HAPs). We used the 'missing mass', defined as the concentration of the measured total particle mass minus the sum of all analyzed species, as an additional variable and implemented an auxiliary equation to constrain the sum of all species mass fractions to be 100%. Regardless of whether the above constraint was implemented and/or the additional VOCs data were included with the PM(2.5) data, the models identified that wood burning (24%-31%), secondary sulfate (20%-24%) and secondary nitrate (15%-20%) were the main contributors to PM(2.5). Using only PM(2.5) data, the model distinguished two diesel features with the 100% constraint, but identified only one diesel feature without the constraint. When both PM(2.5) and VOCs data were used, one additional feature was identified as the major contributor (26%) to total VOC mass. Adding VOCs data to the speciated PM(2.5) data in source apportionment modeling resulted in more accurate source contribution estimates for combustion related sources as evidenced by the less 'missing mass' percentage in PM(2.5). Using the source contribution estimates, we evaluated the validity of using black carbon (BC) as a surrogate for diesel exhaust. We found that BC measured with an aethalometer at 370 nm and 880 nm had reasonable correlations with the estimated concentrations of diesel particulate matters (r>0.7), as well as with the estimated concentrations of wood burning particles during the heating seasons (r=0.56-0.66). This indicates that the BC is not a unique tracer for either source. The difference in BC between 370 and 880 nm, however, correlated well exclusively with the estimated wood smoke source (r=0.59) and may be used to separate wood smoke from diesel exhaust. Thus, when multiple BC related sources exist in the same monitoring environment, additional data processing or modeling of the BC measurements is needed before these measurements could be used to represent the diesel exhaust.     

The RelB-BLNK Axis Determines Cellular Response to a Novel Redox-Active Agent Betamethasone during Radiation Therapy in Prostate Cancer

Aberrant levels of reactive oxygen species (ROS) are potential mechanisms that contribute to both cancer therapy efficacy and the side effects of cancer treatment. Upregulation of the non-canonical redox-sensitive NF-kB family member, RelB, confers radioresistance in prostate cancer (PCa). We screened FDA-approved compounds and identified betamethasone (BET) as a drug that increases hydrogen peroxide levels in vitro and protects non-PCa tissues/cells while also enhancing radiation killing of PCa tissues/cells, both in vitro and in vivo. Significantly, BET increases ROS levels and exerts different effects on RelB expression in normal cells and PCa cells. BET induces protein expression of RelB and RelB target genes, including the primary antioxidant enzyme, manganese superoxide dismutase (MnSOD), in normal cells, while it suppresses protein expression of RelB and MnSOD in LNCaP cells and PC3 cells. RNA sequencing analysis identifies B-cell linker protein (BLNK) as a novel RelB complementary partner that BET differentially regulates in normal cells and PCa cells. RelB and BLNK are upregulated and correlate with the aggressiveness of PCa in human samples. The RelB-BLNK axis translocates to the nuclear compartment to activate MnSOD protein expression. BET promotes the RelB-BLNK axis in normal cells but suppresses the RelB-BLNK axis in PCa cells. Targeted disruptions of RelB-BLNK expressions mitigate the radioprotective effect of BET on normal cells and the radiosensitizing effect of BET on PCa cells. Our study identified a novel RelB complementary partner and reveals a complex redox-mediated mechanism showing that the RelB-BLNK axis, at least in part, triggers differential responses to the redox-active agent BET by stimulating adaptive responses in normal cells but pushing PCa cells into oxidative stress overload.     

Self-concept changes of junior high students.

Previous studies have identified a relationship between self-concept and school behavior. The present study investigated whether self-concept (as measured by Bills's Index of Attitudes and Values) and school behavior of problem junior high school students could be changed by participation in a teacher helper program in an elementary school. Ss matched on grade (8th or 9th) sex, and GPA were randomly assigned to an experimental or a control group (n = 12). Self-concept ratings of experimental Ss who participated in the teacher helper program for 8 wks significantly increased during this period, while controls did not change. Neither group changed significantly in attendance or GPA. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Thymoquinone Suppresses Angiogenesis in DEN-Induced Hepatocellular Carcinoma by Targeting miR-1-3p

Hepatocellular carcinoma (HCC) is characterized by its high vascularity and metastasis. Thymoquinone (TQ), the main bio-active constituent of Nigella sativa, has shown anticancer and hepatoprotective effects. TQ’s anticancer effect is mediated through miRNA regulation. miR-1-3p plays a significant role in various cancers but its role in HCC invasiveness remains poorly understood. Bio-informatics analysis predicted that the 3′-UTR of TIMP3 is a target for miR-1-3p; Rats were equally divided into four groups: Group 1, the negative control; Group 2 received TQ; Group 3 received DEN; and Group 4 received DEN after pretreatment with TQ. The expression of TIMP3, MMP2, MMP9, and VEGF in rats’ liver was determined immunohistochemically. RT-qPCR was used to measure the miR-1-3p level in rats’ liver, and TIMP3, MMP2, MMP9, and VEGF in the HepG2 cells after being transfected with miR-1-3p mimic or inhibitor; In rats pretreated with TQ, a decreased expression of MMP2, MMP9 and VEGF, and increased expression levels of TIMP3 and miR-1-3p were detected. Treating the HepG2 cells with miR-1-3p mimic led to the upregulation of TIMP3 and downregulation of MMP2, MMP9, and VEGF, and showed a significant delay in wound healing; These results suggested that the anti-angiogenic effect of TQ in HCC may be mediated through the regulation of miR-1-3p.