A Rapid, Fluorescence-Based Field Screening Technique for Organic Species in Soil and Water Matrices.

Real-time detection of hydrocarbon contaminants in the environment presents analytical challenges because traditional laboratory-based techniques are cumbersome and not readily field portable. In the current work, a method for rapid and semi-quantitative detection of organic contaminants, primarily crude oil, in natural water and soil matrices has been developed. Detection limits in the parts per million and parts per billion were accomplished when using visual and digital detection methods, respectively. The extraction technique was modified from standard methodologies used for hydrocarbon analysis and provides a straight-forward separation technique that can remove interference from complex natural constituents. For water samples this method is semi-quantitative, with recoveries ranging from 70 % to 130 %, while measurements of soil samples are more qualitative due to lower extraction efficiencies related to the limitations of field-deployable procedures.

Cerebrovascular accident under anesthesia during dental surgery.

Stroke, or cerebrovascular accident (CVA), is a medical emergency that may lead to permanent neurological damage, complications, and death. The rapid loss of brain function due to disruption of the blood supply to the brain is caused by blockage (thrombosis, arterial embolism) or hemorrhage. The incidence of CVA during anesthesia for noncardiac nonvascular surgery is as high as 1% depending on risk factors. Comprehensive preoperative assessment and good perioperative management may prevent a CVA. However, should an ischemic event occur, appropriate and rapid management is necessary to minimize the deleterious effects caused to the patient. This case report describes a patient who had an ischemic CVA while under general anesthesia for dental alveolar surgery and discusses the anesthesia management.

Effect of ion energies on the surface interactions of NO formed in nitrogen oxide plasma systems.

The contributions of various gas-phase species in surface reactions are of significant value to assess and improve catalytic substrates for abatement of vehicular emissions. The impact of ions on surface scatter of NO radicals is investigated with an aim toward improving and tailoring surfaces for the reduction or removal of nitrogen oxide (N(x)O(y)) species via inductively coupled plasmas (ICPs). Nascent ions are monitored via mass spectrometry and energy analysis for a variety of N(x)O(y) precursor gases. The total average ion energy ((total)) determined for all ions within each respective plasma system shows a strong positive correlation with applied rf power and a negative correlation with system pressure. The imaging of radicals interacting with surfaces (IRIS) technique was used to determine the role ions play in the surface scatter of NO radicals. The net effect of ions on substrate processing is largely dependent upon (total). Scatter coefficients (S), determined for ion-limited and ion-rich plasma systems were used to correlate (total) and scatter. The resultant effect is that ions play a substantial role in scatter of NO only when (total) > ~50 eV. The majority of systems studied contained ions below this energy threshold, suggesting knowledge of ion energies is integral to appropriately controlling the chemistry occurring between the gas-phase and surface.

Drug therapy for the pregnant dental patient.

Providing needed dental treatment, managing oral infection, and controlling pain are essential functions of dentists for helping patients maintain overall health during pregnancy. Medications commonly required for dental care consist of local anesthetics and associated vasoconstrictors, centrally and peripherally acting analgesics, sedative and anxiolytic agents, and antibiotics. Therapeutic drugs routinely used in dental practice are selected because of their known safety and effectiveness. However, for a pregnant patient requiring dental care, the agents routinely prescribed should be reevaluated for potential risks to the mother and/or fetus. The decision to administer a specific drug requires that the benefits outweigh the potential risks of the drug therapy. This article reviews and updates the recommendations for using dental therapeutic agents, thereby enabling general practitioners to select the safest drugs when treating pregnant dental patients.

Hepatic Bax inhibitor-1 inhibits IRE1alpha and protects from obesity-associated insulin resistance and glucose intolerance.

The unfolded protein response (UPR) or endoplasmic reticulum (ER) stress response is a physiological process enabling cells to cope with altered protein synthesis demands. However, under conditions of obesity, prolonged activation of the UPR has been shown to have deteriorating effects on different metabolic pathways. Here we identify Bax inhibitor-1 (BI-1), an evolutionary conserved ER-membrane protein, as a novel modulator of the obesity-associated alteration of the UPR. BI-1 partially inhibits the UPR by interacting with IRE1alpha and inhibiting IRE1alpha endonuclease activity as seen on the splicing of the transcription factor Xbp-1. Because we observed a down-regulation of BI-1 expression in liver and muscle of genetically obese ob/ob and db/db mice as well as in mice with diet-induced obesity in vivo, we investigated the effect of restoring BI-1 expression on metabolic processes in these mice. Importantly, BI-1 overexpression by adenoviral gene transfer dramatically improved glucose metabolism in both standard diet-fed mice as well as in mice with diet-induced obesity and, critically, reversed hyperglycemia in db/db mice. This improvement in whole body glucose metabolism and insulin sensitivity was due to dramatically reduced gluconeogenesis as shown by reduction of glucose-6-phosphatase and phosphoenolpyruvate carboxykinase expression. Taken together, these results identify BI-1 as a critical regulator of ER stress responses in the development of obesity-associated insulin resistance and provide proof of concept evidence that gene transfer-mediated elevations in hepatic BI-1 may represent a promising approach for the treatment of type 2 diabetes.

Gas-phase chemistry in inductively coupled plasmas for NO removal from mixed gas systems.

Inductively coupled rf plasmas were used to investigate the removal of NO from a variety of gas mixtures. Laser-induced fluorescence and optical emission spectroscopy were employed to measure the relative gas-phase density of NO as a function of the applied rf power, gas mixture, and catalytic substrate type. In general, the overall density of NO decreases as a function of applied rf power in both NO and N(2)/O(2) plasmas, but the addition of gases such as H(2)O vapor and CH(4), as well as the presence of Au-coated substrates, significantly affects the behavior of NO in these systems. Rotational and vibrational temperatures for NO were measured using laser-induced fluorescence excitation spectra and optical emission spectra. Results show NO vibrational temperatures are about a factor of 5 higher than rotational temperatures and indicate little dependence on applied rf power, feed gas composition, or overall system pressure. Possible mechanisms for the observed changes in [NO] as well as the rotational and vibrational temperature data are addressed.

Bcl-B expression in human epithelial and nonepithelial malignancies.

PURPOSE: Apoptosis plays an important role in neoplastic processes. Bcl-B is an antiapoptotic Bcl-2 family member, which is known to change its phenotype upon binding to Nur77/TR3. The expression pattern of this protein in human malignancies has not been reported. EXPERIMENTAL DESIGN: We investigated Bcl-B expression in normal human tissues and several types of human epithelial and nonepithelial malignancy by immunohistochemistry, correlating results with tumor stage, histologic grade, and patient survival. RESULTS: Bcl-B protein was strongly expressed in all normal plasma cells but found in only 18% of multiple myelomas (n = 133). Bcl-B immunostaining was also present in normal germinal center centroblasts and centrocytes and in approximately half of diffuse large B-cell lymphoma (n = 48) specimens, whereas follicular lymphomas (n = 57) did not contain Bcl-B. In breast (n = 119), prostate (n = 66), gastric (n = 180), and colorectal (n = 106) adenocarcinomas, as well as in non-small cell lung cancers (n = 82), tumor-specific overexpression of Bcl-B was observed. Bcl-B expression was associated with variables of poor prognosis, such as high tumor grade in breast cancer (P = 0.009), microsatellite stability (P = 0.0002), and left-sided anatomic location (P = 0.02) of colorectal cancers, as well as with greater incidence of death from prostate cancer (P = 0.005) and shorter survival of patients with small cell lung cancer (P = 0.009). Conversely, although overexpressed in many gastric cancers, Bcl-B tended to correlate with better outcome (P = 0.01) and more differentiated tumor histology (P < 0.0001). CONCLUSIONS: Tumor-specific alterations in Bcl-B expression may define subsets of nonepithelial and epithelial neoplasms with distinct clinical behaviors.

Certification in Medical Physics Imaging, The Workforce and Training Models.

The pathway to becoming a qualified medical physicist (QMP) in the imaging physics disciplines includes several certification organizations. Imaging QMPs play an essential role in the safe practice of the diagnostic disciplines, and their qualifications are necessary for compliance with federal bodies and professional accreditation organizations. The future demand for imaging QMPs is largely unknown, but professional organizations that represent these groups agree that efforts should be made to increase the number of matriculating trainees. The number of imaging residency programs that provide the necessary professional experience to enter the certification pathway has increased substantially in recent years. Most of these programs follow a traditional academic hospital-based training model, but guidance on program construction from the accrediting body permits flexibility. Existing training models for medical physics imaging also include consortiums of affiliate partners and private consulting service groups. In this article, the authors briefly review the certification pathways for imaging QMPs, workforce estimates, and training models.

A TR3/Nur77 peptide-based high-throughput fluorescence polarization screen for small molecule Bcl-B inhibitors.

Nuclear receptor TR3/Nur77/NR4A1 binds several antiapoptotic Bcl-2-family proteins (Bcl-B, Bcl-2, Bfl-1) in a non-BH3-dependent manner. A 9-amino-acid peptide derived from full-length TR3 with polyarginine tail (TR3-r8) recapitulates TR3's binding specificity, displaying high affinity for Bcl-B. TR3-r8 peptide was used to screen for small molecule Bcl-B inhibitors. A fluorescence polarization assay (FPA) employing fluorescein isothiocyanate (FITC)-labeled TR3-r8 peptide (FITC-TR3-r8) and Bcl-B protein was optimized, with nonfluorescent TR3-r8 serving to demonstrate reversible, competitive binding. Approximately 50,000 compounds were screened at 3.75 mg/L, yielding 145 reproducible hits with > or =50% FITC-TR3-r8 displacement (a confirmed hit rate of 0.29%). After dose-response analyses and counterscreening with an unrelated FITC-based FPA, 6 candidate compounds remained. Nuclear magnetic resonance (NMR) showed that 2 of these compounds bound Bcl-B, but not glutathione S-transferase (GST) control protein. One Bcl-B-binding compound was unable to displace FITClabeled BH3 peptides from Bcl-B, confirming a unique binding mechanism compared with traditional antagonists of antiapoptotic Bcl-2-family proteins. This compound bound Bcl-B with Kd 1.94 +/- 0.38 microM, as determined by isothermal titration calorimetry. Experiments using Bcl-B overexpressing HeLa cells demonstrated that this compound induced Bcl-B-dependent cell death. The current FPA represents a screen that can identify noncanonical inhibitors of Bcl-2-family proteins.

Daxx represses expression of a subset of antiapoptotic genes regulated by nuclear factor-kappaB.

Daxx is a nuclear protein that localizes to PML oncogenic domains, sensitizes cells to apoptosis, and functions as a transcriptional repressor. We found that Daxx represses the expression of several antiapoptotic genes regulated by nuclear factor-kappaB, including cIAP2, in human tumor cell lines. Daxx interacts with RelB and inhibits RelB-mediated transcriptional activation of the human cIAP2 gene promoter. Daxx also forms complexes with RelB while bound to its target sites in the cIAP2 promoter, as shown by electrophoretic mobility shift assays and chromatin immunoprecipitation experiments. Using cells from daxx-/- mouse embryos, we observed that levels of the corresponding murine c-IAP mRNA and protein are increased in cells lacking Daxx. Conversely, c-IAP mRNA and protein levels were reduced in relB-/- cells. Taken together, these observations provide a mechanism that links two previously ascribed functions of Daxx: transcriptional repression and sensitization to apoptosis.

Oxidized analogs of Di(1H-indol-3-yl)methyl-4-substituted benzenes are NR4A1-dependent UPR inducers with potent and safe anti-cancer activity.

Di(1H-indol-3-yl)(4-trifluoromethylphenyl)methane (DIM-Ph-4-CF3) is an analog of orphan nuclear receptor 4A1 (NR4A1) ligand cytosporone B. We have synthesized several oxidation products of DIM-Ph-4-CF3, focusing on analogs with electron-withdrawing or donating groups at their phenyl ring 4-positions, and examined their anti-cancer activity and mechanism-of-action. Mesylates (DIM-Ph-4-X+ OMs-s) having CF3, CO2Me and Cl groups were more effective inhibitors of cancer cell viability than their precursors. 19F NMR spectroscopy and differential scanning calorimetry strongly indicated interactions of DIM-Ph-4-CF3+ OMs- with the NR4A1 ligand binding domain, and compound-induced apoptosis of prostate cancer cells was dependent on NR4A1. DIM-Ph-4-CF3+ OMs- showed robust inhibition of LNCaP prostate cancer xenografts with no apparent toxicity. In vitro and in vivo, DIM-Ph-4-CF3+ OMs- activated proapoptotic unfolded protein response (UPR) signaling in prostate cancer cells. Independently of DIM-Ph-4-CF3+ OMs-, the bulk of NR4A1 localized to the cytoplasm in various cancer cell lines, suggesting a cytoplasmic mechanism-of-action of DIM-Ph-4-CF3+ OMs- in UPR induction and cell death. In summary, the data suggest that oxidized analogs of DIM-Ph-4-CF3 possess potent and safe anti-cancer activity which is mediated through UPR signaling downstream of NR4A1 binding.

An allergic reaction following intramuscular administration of ketamine and midazolam.

A 6-year-old female in good health presented with no known drug allergies for dental treatment under general anesthesia. Following the preoperative evaluation, the patient received intramuscular premedication consisting of midazolam (1 mg) and Ketamine (60 mg) into the left deltoid muscle. During patient transfer, anesthesia personnel detected a hive developing in proximity to the patient's right ear lobe. The subject was directly placed into the operative chair, and a physical exam revealed urticaria on the neck, back, and torso. In addition, an audible wheeze was detected with lung auscultation. Investigations carried out after the incident revealed a positive reaction to ketamine

A weight-of-evidence approach to identify nanomaterials in consumer products: a case study of nanoparticles in commercial sunscreens.

Nanoscale ingredients in commercial products represent a point of emerging environmental concern due to recent findings that correlate toxicity with small particle size. A weight-of-evidence (WOE) approach based upon multiple lines of evidence (LOE) is developed here to assess nanomaterials as they exist in consumer product formulations, providing a qualitative assessment regarding the presence of nanomaterials, along with a baseline estimate of nanoparticle concentration if nanomaterials do exist. Electron microscopy, analytical separations, and X-ray detection methods were used to identify and characterize nanomaterials in sunscreen formulations. The WOE/LOE approach as applied to four commercial sunscreen products indicated that all four contained at least 10% dispersed primary particles having at least one dimension <100 nm in size. Analytical analyses confirmed that these constituents were comprised of zinc oxide (ZnO) or titanium dioxide (TiO2). The screening approaches developed herein offer a streamlined, facile means to identify potentially hazardous nanomaterial constituents with minimal abrasive processing of the raw material.

A high-throughput assay for modulators of ligand-gated chloride channels.

Invertebrate glutamate-gated chloride channels (GluCls) are important targets for anthelmintics and insecticides such as ivermectin. To facilitate screening for novel GluCl modulators, the Caenorhabditis elegans GluCl alpha2beta channel was chosen as a surrogate for parasite channels not yet cloned, and an inducible stable human embryonic kidney cell line was generated. Functional expression of the alpha2 and beta subunits was confirmed by whole-cell voltage clamp assays. Using this cell line, a high-throughput assay was developed that detects membrane potential changes associated with the activation of GluCls. In this assay, membrane depolarization was quantified via changes in fluorescence resonance energy transfer between two membrane-associated dyes. Robust and reproducible signals were detected in response to addition of glutamate or ivermectin. This assay was used for the screening of over 180,000 samples from natural and synthetic sources.

Automated external defibrillator use among the general population.

Automated External Defibrillators (AED) are becoming more prominent in public locations within the mainstream of our society. They are marketed as providing the ability for a broader range of people, beyond clinicians and community emergency medical services personal, to successfully defibrillate a person in cardiac arrest. The objectives of this study were to determine whether or not a member of the general population, without previous exposure to an AED, could successfully operate an AED, thus delivering the necessary shock in ventricular fibrillation arrest. In addition, we analyzed the relationship between health care training and the time required to defibrillate a patient using an AED and investigated the overall success of operating an AED with respect to health care training. Utilizing an AED trainer, we conducted a timed trial study of five subject categories (general population; first-year dental students; third-year dental students; dentists, hygienists, and nurses; and anesthesiologists and surgeons) as each operator attempted to defibrillate a mannequin (n=50). Their times, success in defibrillation, and comments were recorded. The general population group experienced an 80 percent failure rate, while the other groups showed an inverse relationship between failure rates and the amount of health care training. Overall, only 58 percent of the subjects successfully performed the defibrillation with the AED. Operator speed in relation to the amount of health care training showed another inverse relationship as times decreased from group one (general population) to group five (anesthesiologists and surgeons). The findings suggest that prior exposure to an AED leads to a greater number of successful defibrillations. It remains unclear at this time as to whether a member of the general population can successfully operate an AED.

Determination of isoelectric points and the role of pH for common quartz crystal microbalance sensors.

Isoelectric points (IEPs) were determined by the method of contact angle titration for five common quartz crystal microbalance (QCM) sensors. The isoelectric points range from mildly basic in the case of Al2O3 sensors (IEP = 8.7) to moderately acidic for Au (5.2) and SiO2 (3.9), to acidic for Ag (3.2) and Ti (2.9). In general, the values reported here are indicative of inherent surface oxides. A demonstration of the effect of the surface isoelectric point on the packing efficiency of thin mucin films is provided for gold and silica QCM sensors. It is determined that mucin layers on both substrates achieve a maximum and equal layer density of ∼3500 kg/m(3) at the corresponding IEP of either QCM sensor. This implies that mucin film packing is dependent upon short-range electrostatic interactions at the sensor surface.

Interleukin-1 receptor-associated kinase-2 (IRAK2) is a critical mediator of endoplasmic reticulum (ER) stress signaling.

Endoplasmic reticulum (ER) stress occurs when unfolded proteins accumulate in the lumen of the organelle, triggering signal transduction events that contribute either to cellular adaptation and recovery or alternatively to cellular dysfunction and death. ER stress has been implicated in numerous diseases. To identify novel modulators of ER stress, we undertook a siRNA library screen of the kinome, revealing Interleukin-1 Receptor-Associated Kinase-2 (IRAK2) as a contributor to unfolded protein response (UPR) signaling and ER stress-induced cell death. Knocking down expression of IRAK2 (but not IRAK1) in cultured mammalian cells suppresses ER stress-induced expression of the pro-apoptotic transcription factor CHOP and activation of stress kinases. Similarly, RNAi-mediated silencing of the IRAK family member Tube (but not Pelle) suppresses activation of stress kinase signaling induced by ER stress in Drosophila cells. The action of IRAK2 maps to the IRE1 pathway, rather than the PERK or ATF6 components of the UPR. Interestingly, ER stress also induces IRAK2 gene expression in an IRE1/XBP1-dependent manner, suggesting a mutually supporting amplification loop involving IRAK2 and IRE1. In vivo, ER stress induces Irak2 expression in mice. Moreover, Irak2 gene knockout mice display defects in ER stress-induced CHOP expression and IRE1 pathway signaling. These findings demonstrate an unexpected linkage of the innate immunity machinery to UPR signaling, revealing IRAK2 as a novel amplifier of the IRE1 pathway.

Contributions of CF and CF2 Species to fluorocarbon film composition and properties for C(x)F(y) plasma-enhanced chemical vapor deposition.

Inductively-coupled C(x)F(y) (y/x = 2.0-4.0) plasma systems were investigated to determine relationships between precursor chemistry, CF(n) radical-surface reactivities, and surface properties of deposited films. The contributions of CF(n) (n = 1, 2) radicals to film properties were probed via gas-phase diagnostics and the imaging of radicals interacting with surfaces (IRIS) technique. Time-resolved radical emission data elucidate CF(g) and CF(2)(g) production kinetics from the C(x)F(y) source gases and demonstrate that CF(4) plasmas inherently lag in efficacy of film formation when compared to C(2)F(6), C(3)F(8), and C(3)F(6) systems. IRIS data show that as the precursor y/x ratio decreases, the propensity for CF(n) scatter concomitantly declines. Analyses of the composition and characteristics of fluorocarbon films deposited on Si wafers demonstrate that surface energies of the films decrease markedly with increasing film fluorine content. In turn, increased surface energies correspond with significant decreases in the observed scatter coefficients for both CF and CF(2). These data improve our molecular-level understanding of CF(n) contributions to fluorocarbon film deposition, which promises advancements in the ability to tailor FC films to specific applications.

TR-FRET-based high-throughput screening assay for identification of UBC13 inhibitors.

UBC13 is a noncanonical ubiquitin conjugating enzyme (E2) that has been implicated in a variety of cellular signaling processes due to its ability to catalyze formation of lysine 63-linked polyubiquitin chains on various substrates. In particular, UBC13 is required for signaling by a variety of receptors important in immune regulation, making it a candidate target for inflammatory diseases. UBC13 is also critical for double-strand DNA repair and thus a potential radiosensitizer and chemosensitizer target for oncology. The authors developed a high-throughput screening (HTS) assay for UBC13 based on the method of time-resolved fluorescence resonance energy transfer (TR-FRET). The TR-FRET assay combines fluorochrome (Fl)-conjugated ubiquitin (fluorescence acceptor) with terbium (Tb)-conjugated ubiquitin (fluorescence donor), such that the assembly of mixed chains of Fl- and Tb-ubiquitin creates a robust TR-FRET signal. The authors defined conditions for optimized performance of the TR-FRET assay in both 384- and 1536-well formats. Chemical library screens (total 456 865 compounds) were conducted in high-throughput mode using various compound collections, affording superb Z' scores (typically >0.7) and thus validating the performance of the assays. Altogether, the HTS assays described here are suitable for large-scale, automated screening of chemical libraries in search of compounds with inhibitory activity against UBC13.