Search

Cardiovascular disease remains the top cause of morbidity and mortality in the United States. Atherosclerotic plaques are known to start in adolescence, and, therefore, young adults can be affected by coronary artery disease. Children with known risk factors, such as genetic predisposition, including familial hyperlipidemias, diabetes, and renal diseases, are at higher risk. With childhood obesity becoming an epidemic in certain parts of the United States, this problem is further highlighted as an important issue affecting children’s health. There are unclear recommendations for pediatricians regarding cholesterol screening of pediatric populations, when to initiate hyperlipidemia treatment with statin therapy, and when to refer to a specialist for further management. This article reviews the epidemiology and pathophysiology of hyperlipidemia, recommendations for screening and types of screening, management (including pharmacology), prognosis, and prevention.

Nuclear waste management facilities at Chalk River Laboratories (CRL) use below-ground “tile-holes” to store solid waste from various activities such as medical isotope production. After long periods of isotopic decay, some of the waste has decayed down to low activities and can be transferred to low-level waste storage facilities. This paper presents a method to verify the radiation level of the waste inside tile-holes by performing gamma radiation scans along the depth of waste storage tile-holes. Such measurements allow for noninvasive verification of tile-hole contents and provide input to the assessment of radiological risk associated with removal of the waste. Using the radiation profile system, the radiation level of the radioactive waste may be identified based on the radiation profile. This information will support planning for possible transfer of this waste to a licensed waste storage facility designed for low-level waste, thus freeing storage space for possible tile-hole re-use for more highly radioactive waste. CRL-developed small diode-based gamma radiation sensors have been used in these radiation scans. The diode sensors were deployed into verification tubes adjacent to the tile-holes to measure the radiation profile. Over 10 tile-holes have been scanned using this technique since 2009.

Advances in radio astronomy are allowing for deeper and wider areas of the sky to be observed than ever before. Source counts of future radio surveys are expected to number in the tens of millions. Source finding techniques are used to identify sources in a radio image, however, these techniques identify single distinct sources and are challenged to identify multi-component sources, that is to say, where two or more distinct sources belong to the same underlying physical phenomenon, such as a radio galaxy. Identification of such phenomena is an important step in generating catalogues from surveys on which much of the radio astronomy science is based. Historically, identifying multi-component sources was conducted by visual inspection, however, the size of future surveys makes manual identification prohibitive. An algorithm to automate this process using statistical techniques is proposed. The algorithm is demonstrated on two radio images. The output of the algorithm is a catalogue where nearest neighbour source pairs are assigned a probability score of being a component of the same physical object. By applying several selection criteria, pairs of sources which are likely to be multi-component sources can be determined. Radio image cutouts are then generated from this selection and may be used as input into radio source classification techniques. Successful identification of multi-component sources using this method is demonstrated.

Maca (Lepidium meyenii, synonym L. peruvianum) was analyzed using a systematic approach employing principal component analysis of flow injection mass spectrometry fingerprints (no chromatographic separation) to guide the selection of samples for metabolite profiling and DNA next generation sequencing. Samples consisted of 39 commercial maca supplements from 11 manufacturers, 31 unprocessed maca tubers grown in Peru and China, and a historic non-tuber maca sample from Peru. Principal component analysis of flow injection mass spectrometry fingerprints initially placed all the maca samples in three classes with similar chemical composition: commercial maca samples, tubers grown in Peru, and tubers grown in China. Metabolite profiling identified 67 compounds in the negative mode and 51 compounds in the positive mode. Compounds identified by metabolite profiling (macamides, glucosinolates, amino acids, fatty acids, polyunsaturated fatty acids, saccharides, imidazoles) were then used to identify ions in the flow injection mass spectrometry fingerprints. The tuber fingerprints were analyzed by factorial multivariate analysis of variance revealing that black, red, and yellow maca from Peru and black and yellow maca from China were compositionally different with respect to color and country. Critical ions were identified that allowed for the differentiation of maca between colors from the same country or between two countries with the same color. Genetically, all samples were confirmed to be L. meyenii based on next generation sequencing at three gene regions (ITS2, psbA, and trnL) and comparison to recorded sequences of vouchered standards.

Background: Distal radius fractures represent a common fracture pattern frequently treated with volar locked plating for fixation. However, other methods may provide equivalent outcomes and minimize risks associated with the volar approach and hardware placement. One such method is the radial plate. Methods: After confirmation of institutional board review, we retrospectively reviewed data from 7 patients with our primary functional outcomes measured by Mayo wrist and Disabilities of the Arm, Shoulder and Hand (QuickDASH) scores. The most recent radiographic and motion parameters were recorded. Results: Mean age at time of injury was 49 years (range, 19-68 years) with clinical follow-up of 81.6 months (range, 43.5-95.5 months). Five patients had good to excellent outcomes with a mean QuickDASH score of 0.92 for those patients. The mean QuickDASH score for all patients was 18.5. Mean radial height, inclination, and volar tilt were within 5% of the contralateral side. Standard deviation values for radiographic measures and clinical range of motion indicate significant variability in our data set. Conclusions: Although the results are mixed, our small cohort indicates radial plate fixation could provide a viable alternative to volar plate fixation of distal radius fractures. Further prospective investigation is warranted to better describe long-term outcomes using this technique.

While considerable research has targeted gait, balance and preventing falls in individuals with Parkinson’s disease (PD), less in known about the ability to rise from the floor in this population. The aims of this study were to 1) Examine the relationship between locomotion and physical performance tests and the timed supine to stand performance measure and to 2) Identify both the time required and predominant motor patterns utilized by persons with PD to complete to floor rise transition. A cross-sectional design was utilized. Twenty community-dwelling older adults with PD (mean age 74.8+/-9.5 years; 13 men) performed a standardized floor rise test and locomotion tests in a structured task circuit. Subject demographic and anthropometric data were also collected. Statistical analyses included descriptive statistics and Pearson Product Moment correlations. Fifteen subjects (75%) demonstrated the crouch kneel pattern and fourteen (70%) used an all-4’s strategy to rise to stand. The mean time to rise from the floor was 14.9 (+/- 7.6) seconds and slower than published norms for persons without PD. Nine subjects required the use of a chair to perform floor recovery. Supine to stand performance time was significantly correlated with the: Dynamic Gait Index (r= - 0.66; p

In the setting a near or complete upper extremity amputations with significant soft tissue loss and neurovascular compromise, upper extremity surgeons are faced with the challenge of limb salvage. There are a multitude of treatment options for managing skeletal and soft tissue injuries including provisional fixation, staged reconstruction, and an acute shortening osteotomy with primary rigid internal fixation. However, many complications are associated with these techniques. Complications of provisional fixation include pin tract infection and loosening, tethering of musculotendinous units, nonunion, and additional surgeries. Staged reconstruction includes a variety of techniques: distraction osteogenesis, bone transport, or vascularized and non-vascularized structural autograft or allograft, but the risks often outweigh the benefits. Risks include nonunion, postoperative vascular complications necessitating reoperation, and the inability to return to the previous level of function at an average of 24 months. Acute shortening osteotomy with internal fixation offers the advantage of a single-stage procedure that provides for decreasing the soft tissue loss, provides a rigid platform to protect the delicate neurovascular repair, and alleviates unwanted tension at the repair sites. This review discusses the literature on the surgical treatment of severe upper extremity trauma with associated neurovascular injury over the past 75 years, and aims to evaluate the indications, surgical techniques, clinical and functional outcomes, and complications associated with acute shortening osteotomy with rigid internal fixation. Although this technique is not without risks, it is well-tolerated in the acute setting with a complication profile comparable to other techniques of fixation while remaining a single procedure.

The modeling of biological data can be carried out using structured sound and musical process in conjunction with integrated visualizations. With a future goal of improving the speed and accuracy of techniques currently in use for the production of synthetic high value chemicals through the greater understanding of data sets, the madBPM project couples real-time audio synthesis and visual rendering with a highly flexible data-ingestion engine. Each component of the madBPM system is modular, allowing for customization of audio, visual and data-based processing.

Twenty years of professionalism has seen rugby union undergo dramatic transformations, from changes to everyday training cultures to the growth of the Rugby World Cup into one of the largest global sporting events.The Rugby World in the Professional Erais the first book to examine the effect that professionalism has had across a number of different aspects of the game and the wider socio-cultural significance of these changesthrough case studiesfrom across the globe. Drawing on contributions from scholars from across the rugby-playing world, the book explores the role of rugby's professionalisation through a number of social-scientific lenses, including: labour migration race and indigenous populations the globalisation of the game mega-event management male sexualities media representations of rugby - from broadcasting matches to rugby in museums andon stage and screenOffering insights into under-researched areas of the sport, such as the growth of Rugby Sevens into an Olympic sport, and providing the most up-to-date recent history of the sport available, The Rugby World in the Professional Erais essential reading for anyone with an academic interest in rugby, and any student or scholar with interests in sports history, sports sociology, sport managementor the economics of professional sport.

Plastic labware is used in all processes of modern pharmaceutical research, including compound storage and biological assays. The use of these plastics has created vast increases in productivity and cost savings as experiments moved from glass test tubes and capillary pipettes to plastic microplates and multichannel liquid handlers. One consequence of the use of plastic labware, however, is the potential release of contaminants and their resultant effects on biological assays. We report herein the identification of biologically active substances released from a commonly used plastic microplate. The active contaminants were identified by gas chromatography-mass spectroscopy as dodecan-1-ol, dodecyl 3-(3-dodecoxy-3-oxopropyl)sulfanylpropanoate, and dodecanoic acid, and they were found to be selective monoamine oxidase-B inhibitors.

This paper proposes methods for generation and implementation of uniform, large-scale data from auralized MIDI music files for use with deep learning networks for polyphonic pitch perception and impulse response recognition. This includes synthesis and sound source separation of large batches of multitrack MIDI files in non-real time, convolution with artificial binaural room impulse responses, and techniques for neural network training. Using ChucK, individual tracks for each MIDI file, containing the ground truth for pitch and other parameters, are processed concurrently with variable Synthesis ToolKit (STK) instruments, and the audio output is written to separate wave files in order to create multiple incoherent sound sources. Then, each track is convolved with a measured or synthetic impulse response that corresponds to the virtual position of the instrument in the room before all tracks are digitally summed. The database now contains the symbolic description in the form of MIDI commands and the auralized music performances. A polyphonic pitch model based on an array of autocorrelation functions for individual frequency bands is used to train a neural network and analyze the data [Work supported by IBM AIRC grant and NSF BCS-1539276.]This paper proposes methods for generation and implementation of uniform, large-scale data from auralized MIDI music files for use with deep learning networks for polyphonic pitch perception and impulse response recognition. This includes synthesis and sound source separation of large batches of multitrack MIDI files in non-real time, convolution with artificial binaural room impulse responses, and techniques for neural network training. Using ChucK, individual tracks for each MIDI file, containing the ground truth for pitch and other parameters, are processed concurrently with variable Synthesis ToolKit (STK) instruments, and the audio output is written to separate wave files in order to create multiple incoherent sound sources. Then, each track is convolved with a measured or synthetic impulse response that corresponds to the virtual position of the instrument in the room before all tracks are digitally summed. The database now contains the symbolic description in the form of MIDI commands and the aural...

Larger size and clear cell histopathology are associated with worse outcomes for malignant renal tumors treated with radio frequency ablation. We hypothesize that greater tumor enhancement may be a risk factor for radio frequency ablation failure due to increased vascularity.A retrospective review of patients who underwent radio frequency ablation for renal tumors with contrast enhanced imaging available was performed. The change in Hounsfield units (HU) of the tumor from the noncontrast phase to the contrast enhanced arterial phase was calculated. Radio frequency ablation failure rates for biopsy confirmed malignant tumors were compared using the chi-squared test. Multivariate logistic analysis was performed to assess predictive variables for radio frequency ablation failure. Disease-free survival was calculated using Kaplan-Meier analysis.A total of 99 patients with biopsy confirmed malignant renal tumors and contrast enhanced imaging were identified. The incomplete ablation rate was significantly lower for tumors with enhancement less than 60 vs 60 HU or greater (0.0% vs 14.6%, p=0.005). On multivariate logistic regression analysis tumor enhancement 60 HU or greater (OR 1.14, p=0.008) remained a significant predictor of incomplete initial ablation. The 5-year disease-free survival for size less than 3 cm was 100% vs 69.2% for size 3 cm or greater (p0.01), while 5-year disease-free survival for HU change less than 60 was 100% vs 92.4% for HU change 60 or greater (p=0.24).Biopsy confirmed malignant renal tumors, which exhibit a change in enhancement of 60 HU or greater, experience a higher rate of incomplete initial tumor ablation than tumors with enhancement less than 60 HU. Size 3 cm or greater portends worse 5-year disease-free survival after radio frequency ablation. The degree of enhancement should be considered when counseling patients before radio frequency ablation.

Biological characteristics of ice-associated algal communities were studied in Darnley Bay (western Canadian Arctic) during a 2-week period in July 2008 when the landfast ice cover had reached an advanced stage of melt. We found two distinct and separate algal communities: (1) an interior ice community confined to brine channel networks beneath white ice covers; and (2) an ice melt water community in the brackish waters of both surface melt ponds and the layer immediately below the ice cover. Both communities reached maximum chlorophyll a concentrations of about 2.5 mg m−3, but with diatoms dominating the interior ice while flagellates dominated the melt water community. The microflora of each community was diverse, containing both unique and shared algal species, the latter suggesting an initial seeding of the ice melt water by the bottom ice community. Absorption characteristics of the algae indicated the presence of mycosporine-like amino acids (MAAs) and carotenoid pigments as a photoprotective strategy against being confined to high-light near-surface layers. Although likely not contributing substantially to total annual primary production, these ice-associated communities may play an important ecological role in the Arctic marine ecosystem, supplying an accessible and stable food source to higher trophic levels during the period of ice melt.

Quantification of small molecules using liquid chromatography/tandem mass spectrometry (LC/MS/MS) on a triple quadrupole mass spectrometer has become a common practice in bioanalytical support of in vitro adsorption, distribution, metabolism and excretion (ADME) screening. The bioanalysis process involves primarily three indispensable steps: MS/MS optimization for a large number of new chemical compounds undergoing various screening assays in early drug discovery, high-throughput sample analysis with LC/MS/MS for those chemically diverse compounds using the optimized MS/MS conditions, and post-acquisition data review and reporting. To improve overall efficiency of ADME bioanalysis, an integrated system was proposed featuring an automated and unattended MS/MS optimization, a staggered parallel LC/MS/MS for high-throughput sample analysis, and a sophisticated software tool for LC/MS/MS raw data review as well as biological data calculation and reporting. The integrated platform has been used in bioanalytical support of a serum protein binding screening assay with high speed, high capacity, and good robustness. In this new platform, a unique sample dilution scheme was also introduced. With this dilution design, the total number of analytical samples was reduced; therefore, the total operation time was reduced and the overall throughput was further improved. The performance of the protein binding screening assay was monitored with two controls representing high and low binding properties and an acceptable inter-assay consistency was achieved. This platform has been successfully used for the determination of serum protein binding in multiple species for more than 4000 compounds.

OBJECTIVES/SPECIFIC AIMS: “Cure Quest” is an adventure quest game for mobile tablets that aims to teach the player about the complexities of discovery and development of new medicines. The game instills a sense of wonderment into the learning process, taking the player to a world of magic where a mysterious condition has affected the land and they must follow the steps of the discovery and development process to find a treatment. METHODS/STUDY POPULATION: The game is being developed through a collaboration between faculty and students at ISMMS and the Games and Simulation Arts and Science Program at Rensselaer Polytechnic Institute. The first target audience is 2nd–3rd year medical students, with the future goal of adapting the game to a broader population. RESULTS/ANTICIPATED RESULTS: The game is currently in development, but the project has yielded insight into the design process for serious games in medicine. We found that for a game of this type it is essential not just to have both designers and subject matter experts, but to enable cross-pollination of modes of thinking. Through multiple design iterations and focus groups, we found that a game design approach rooted in narrative and allegorical abstraction would have a better ability to engage the target audience than one focused only on realistic simulation. When complete, we anticipate that the game will improve understanding of the core concepts in drug discovery. DISCUSSION/SIGNIFICANCE OF IMPACT: If successful, the game-based learning approach can help fill key gaps in current formal medical and scientific training, as well as gaps in understanding among the general public. The design process serves as an informative model of evolving collaborative team science.

Salvia divinorum is a natural occurring hallucinogen that is traditionally used by the Mazatec Indians of central Mexico. The diterpene salvinorin A was identified as an active component of S. divinorum over 20 years ago, but only recently has biochemical screening indicated that a molecular target of salvinorin A in vitro is the kappa-opioid receptor. We have examined whether salvinorin A, the C2-substituted derivative salvinorinyl-2-propionate, and salvinorin B can act as kappa-opioid receptor agonists in vivo. We found that following intracerebroventricular injection over a dose range of 1 to 30 microg of both salvinorin A and salvinorinyl-2-propionate produces antinociception in wild-type mice but not in a novel strain of kappa-opioid receptor knockout mice. Moreover, both salvinorin A and salvinorinyl-2-propionate reduce rectal body temperature, similar to conventional kappa-opioid receptor agonists, in a genotype-dependent manner. In addition, we determined that salvinorin A has high affinity for kappa 1- but not kappa 2-opioid receptors, demonstrating selectivity for this receptor subclass. Finally, treatment over the same dose range with salvinorin B, which is inactive in vitro, produced neither antinociceptive nor hypothermic effects in wild-type mice. These data demonstrate that salvinorin A is the active component of S. divinorum, selective for kappa(1)-opioid receptors, and that salvinorin A and specific structurally related analogs produce behavioral effects that require the kappa-opioid receptor.

A simple and efficient synthesis of 3,5‐dibromo‐O‐methyl‐L‐tyrosine is described by direct bromination of commercial O‐methyl‐L‐tyrosine with bromine in aqueous hydrochloric acid. MO calculations are presented to explain a difference in the degree of substitution of O‐methyl‐L‐tyrosine in different solvents.

Following up on our previous findings that the skin possesses steroidogenic activity from progesterone, we now show widespread cutaneous expression of the full cytochrome P450 side-chain cleavage (P450scc) system required for the intracellular catalytic production of pregnenolone, i.e. the genes and proteins for P450scc enzyme, adrenodoxin, adrenodoxin reductase and MLN64. Functionality of the system was confirmed in mitochondria from skin cells. Moreover, purified mammalian P450scc enzyme and, most importantly, mitochondria isolated from placenta and adrenals produced robust transformation of 7-dehydrocholesterol (7-DHC; precursor to cholesterol and vitamin D3) to 7-dehydropregnenolone (7-DHP). Product identity was confirmed by comparison with the chemically synthesized standard and chromatographic, MS and NMR analyses. Reaction kinetics for the conversion of 7-DHC into 7-DHP were similar to those for cholesterol conversion into pregnenolone. Thus, 7-DHC can form 7-DHP through P450scc side-chain cleavage, which may serve as a substrate for further conversions into hydroxy derivatives through existing steroidogenic enzymes. In the skin, 5,7-steroidal dienes (7-DHP and its hydroxy derivatives), whether synthesized locally or delivered by the circulation, may undergo UVB-induced intramolecular rearrangements to vitamin D3-like derivatives. This novel pathway has the potential to generate a variety of molecules depending on local steroidogenic activity and access to UVB.

In recent years β-amino acids have increased their importance enormously in defining secondary structures of β-peptides. Interest in β-amino acids raises the question: Why and how did nature choose α-amino acids for the central role in life? In this article we present experimental results of MS and 31P NMR methods on the chemical behavior of N-phosphorylated α-alanine, β-alanine, and γ-amino butyric acid in different solvents. N-Phosphoryl α-alanine can self-assemble to N-phosphopeptides either in water or in organic solvents, while no assembly was observed for β- or γ-amino acids. An intramolecular carboxylic–phosphoric mixed anhydride (IMCPA) is the key structure responsible for their chemical behaviors. Relative energies and solvent effects of three isomers of IMCPA derived from α-alanine (2a–c), with five-membered ring, and five isomers of IMCPA derived from β-alanine (4a–e), with six-membered ring, were calculated with density functional theory at the B3LYP/6-31G** level. The lower relative energy (3.2 kcal/mol in water) of 2b and lower energy barrier for its formation (16.7 kcal/mol in water) are responsible for the peptide formation from N-phosphoryl α-alanine. Both experimental and theoretical studies indicate that the structural difference among α-, β-, and γ-amino acids can be recognized by formation of IMCPA after N-phosphorylation. © 2003 Wiley Periodicals, Inc. Int J Quantum Chem 94: 232–241, 2003

Introduction: Chronic graft versus host disease (cGVHD) remains a major source of morbidity and mortality following allogeneic transplantation. While corticosteroids remain first line therapy for cGVHD, they are associated with significant toxicity, and a substantial proportion of patients fail to completely respond. Treatments for steroid-refractory cGVHD are limited. While the pathophysiology of chronic GVHD is complex, activated T cells play a critical role, driven by allo-antigen stimulation. As such, inhibition of T cell activation via blockade of co-stimulation has potential as a therapeutic target in cGVHD. Abatacept is a recombinant fusion protein consisting of the extracellular domain of human CTLA-4 and a fragment of the Fc domain of human IgG1 that has been modified to prevent complement fixation and antibody-dependent cellular cytotoxicity. Abatacept is the first drug in a class of agents termed "selective co-stimulation modulators." The CTLA-4 moiety of Abatacept binds specifically to CD80 and CD86 and down-modulates the CD28-mediated co-stimulation of T cells. We conducted a phase I clinical trial was conducted to evaluate the safety, clinical and immune effects of Abatacept in patients with steroid-refractory cGVHD. Methods: The study followed a 3+3 design with two escalating doses of Abatacept to determine the maximum tolerated dose (MTD): 3 mg/kg and 10 mg/kg. Dose-limiting toxicities (DLTs) were defined as Grade 3 or 4 toxicities judged to be probably or definitely related to Abatacept. Infection was not considered a DLT. Abatacept was administered for a total of 6 doses. Doses 1-3 were administered at two-week intervals. One month following Dose 3, Abatacept was given at four-week intervals for three doses (Doses 4-6). Inclusion criteria included recipients of allogeneic bone marrow or stem cell transplantation with myeloablative or reduced intensity conditioning, with cGVHD defined by NIH consensus criteria. Patients must have had treatment with ≥ 0.5 mg/kg/day of prednisone for at least 4 weeks. Patients with active malignant disease relapse or other active malignancy and patients with uncontrolled infection were excluded. Peripheral blood was drawn prior to each dose of Abatacept and following completion of therapy to assess the effect of treatment on circulating T cells. PD-1 expression on circulating T cells, and T cell expression of interferon gamma versus IL-10 was assessed by multichannel FACS analysis. Results: 17 subjects were treated. Three patients were treated at a dose of 3 mg/kg without DLT. Three evaluable patients completed treatment on cohort 2, at a dose of 10mg/kg without DLT. A forth participant withdrew consent following one dose of treatment and therefore is not evaluable. Ten patients were treated on an expansion cohort at a dose of 10mg/kg. We observed one grade 4 pulmonary infection, and three grade 3 pulmonary infections which resolved. Other Abatacept related adverse events included grade 2 gastritis (n=1), grade 2 pain (n=1), and grade 1 diarrhea (n=2), fatigue (n=2), rash (n=1), and skin pain (n=1). Of the 16 evaluable patients, 7 (44%) achieved a clinical partial response as defined by improvement of two disease systems based on the 2011 NIH consensus criteria. Abatacept resulted in a 51.3% reduction in prednisone usage in clinical responders with a mean baseline dose of 27mg compared to a mean dose of 14mg 1 month following the 6th dose of Abatacept (p = 0.01). PD-1 expression on circulating CD4+ and CD8+ T cells increased from a mean of 3.4% and 2.7% respectively at baseline to a mean of 8.9% and 7.6% respectively at one month following the 6thdose of Abatacept in clinical responders (n=3; p Conclusion: Abatacept is well-tolerated in the treatment of steroid-refractory cGVHD. Abatacept resulted in the improvement in NIH cGVHD scores in 44% of patients with steroid-refractory GVHD with a significant decrease in prednisone dose. An increase in PD-1 expression and a skewing toward Th2 cytokines was observed in clinical responders. Based on this promising data, a phase II trial is being initiated. Disclosures Soiffer: GentiumSpA/Jazz Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees. Arnason:Gilead: Consultancy. Avigan:Astex: Research Funding; DCPrime: Research Funding. Rosenblatt:Astex: Research Funding; BMS: Research Funding; DCPrime: Research Funding.

Background: High-resolution MS (HRMS) has recently received a considerable interest in quantitative bioanalysis using full-scan acquisition mode. The benefits include complete elimination of compound-specific MS method development, and simultaneous collection of mass spectral data on both targeted and non-targeted components. One additional advantage that has not been widely discussed is its suitability for simultaneous quantitation of, theoretically, an unlimited number of compounds, which is not possible with selected reaction monitoring (SRM) on a triple quadrupole mass spectrometer. Materials & Methods: We took advantage of this unique bioanalytical capability of HRMS and developed a novel in vitro ADME workflow of cassette incubation of as many as 32 compounds, followed by quantitative bioanalysis using full-scan acquisition on an Orbitrap HRMS. The workflow was evaluated for a serum protein-binding assay and a parallel artificial membrane permeability (PAMPA) assay. Results: The bioanalytical assay displayed acceptable sensitivity, selectivity and linearity for all compounds in the cassettes, and the biological results obtained using this approach were similar to those from discrete incubation and analysis, demonstrating the feasibility of the workflow. Additional benefits of this platform include a saving of analysis time due to the reduced sample numbers from the cassette approach, as well as cost saving due to the reduction in the required assay reagents. Conclusion: Cassette incubation with bioanalysis using HRMS is a feasible approach for high-throughput in vitro ADME assays evaluated in this study.

Solar domestic hot water (SDHW) can be used to reduce energy bills and greenhouse gas emissions associated with heating domestic water. However, one of the most significant barriers to further deployment of solar thermal applications is the space and weight required for storage of the energy collected. Phase change materials (PCMs) are advantageous for daily energy storage with SDHW due to their high storage density and isothermal operation during phase transitions, and would overcome these obstacles. The aim of this paper is to outline the initial steps in the development of a SDHW energy storage system using PCMs, with emphasis on the numerical and experimental studies used to access the phase change and thermal behaviour of the selected PCM. Lauric acid was selected as the PCM based on the melting temperature range which was targeted by studying solar data from an existing solar hot water system in Halifax, Nova Scotia, Canada. Due to the low thermal conductivity of PCMs, additional work is required to develop and validate a design to enhance heat transfer to the storage material using fins. The selected design will be built and installed in an existing large scale solar thermal system on an apartment building in Halifax. The system will be instrumented in order to acquire continuous data (temperatures, flow rates, pressures, etc.) to fully characterize the system.

[1] Melt pond covered sea ice is a ubiquitous feature of the summertime Arctic Ocean when meltwater collects in lower-lying areas of ice surfaces. Horizontal transects were conducted during June 2008 above and below landfast sea ice with melt ponds to characterize surface and bottom topography together with variations in transmitted spectral irradiance. We captured a rapid progression from a highly flooded sea ice surface with lateral drainage toward flaws and seal breathing holes to the formation of distinct melt ponds with steep edges. As the mass of the ice cover decreased due to meltwater drainage and rose upward with respect to the seawater level, the high-scattering properties of ice above the water level (i.e., white ice) were continuously regenerated, while pond waters remained transparent compared to underlying ice. The relatively stable albedos observed throughout the study, even as ice thickness decreased, were directly related to these surface processes. Transmission through the ice cover of incident irradiance in the 400–700 nm wave band ranged from 38% to 67% and from 5% to 16% beneath ponded and white ice, respectively. Our results show that this transmission varied not only as a function of surface type (melt ponds or white ice) areal coverage but also in relation to ice thickness and proximity to other surface types through the influence of horizontal spreading of light. Thus, in contrast to albedo, this implies that regional transmittance estimates need to consider melt pond size and shape distributions and variations in optical properties and thickness of the ice cover.

Heavy metal contamination of herbal products is a major concern in the herbal and dietary supplement industry. Heavy metal exposure is well-documented to cause a variety of adverse human health effects and to negatively impact our environment. The Final Rule for Dietary Supplements for current good manufacturing practice regulation, 21 U.S. Food and Drug Administration Code of Federal Regulations 111, requires dietary supplement manufacturers to establish herbal purity limits for heavy metal contaminants considered safe for human consumption. Heavy metals may enter into the herbal manufacturing process via bioaccumulation from the harvest site or during postharvest processing phases, such as drying and/or liquid extraction. Traditionally, herbalists have used hydroethanolic solvents to extract herbal biomasses in pure food-grade ethanolwater combinations with solvent polarities capable of removing a wide range of hydrophilic and lipophilic constituents. The presented data demonstrate that hydroethanolic solvents are not completely efficient in the extraction of heavy metal accumulations from plant matrixes; and can act as an effective decontamination step in herbal product processing.

In order to identify potential cytochrome P-450 3A4 (drug-metabolizing enzyme) inducers at an early stage of the drug discovery process, a cell-based transactivation high-throughput luciferase reporter assay for the human pregnane X receptor (PXR) in HepG2 cells has been implemented and multiplexed with a viability end point for data interpretation, as part of a Lead Profiling portfolio of assays. As a routine part of Lead Profiling operations, assays are periodically evaluated for utility as well as for potential improvements in technology or process. We used a recent evaluation of our PXR-transactivation assay as a model for the application of Lean Thinking-based process analysis to lab-bench assay optimization and automation. This resulted in the development of a 384-well multiplexed homogeneous assay simultaneously detecting PXR transactivation and HepG2 cell cytotoxicity. In order to multiplex fluorescent and luminescent read-outs, modifications to each assay were necessary, which included optimization of multiple assay parameters such as cell density, plate type, and reagent concentrations. Subsequently, a set of compounds including known cytotoxic compounds and PXR inducers were used to validate the multiplexed assay. Results from the multiplexed assay correlate well with those from the singleplexed assay formats measuring PXR transactivation and viability separately. Implementation of the multiplexed assay for routine compound profiling provides improved data quality, sample conservation, cost savings, and resource efficiencies.

Novel semisynthetic analogs of salvinorin A, a full agonist having extraordinary affinity as well as selectivity for the κ-opioid receptor (KOR), were obtained in good yields. The derivatives are remarkable for their unusual and unique hemiacetal structure in the salvinorin series of compounds. The formation of the hemiacetal occurs with epimerization at C-12, thus preserving the original configuration of salvinorin A. The dimethyl ester derivative of the hemiacetal was found to have an affinity for both KOR and MOR (μ-opioid receptor).

Cryopreserved, transiently transfected HepG2 cells were compared to freshly transfected HepG2 cells for use in a pregnane X receptor (PXR) transactivation assay. Assay performance was similar for both cell preparations; however, cryopreserved cells demonstrated less interassay variation. Validation with drugs of different PXR activation potencies and efficacies demonstrated an excellent correlation (r2 > 0.95) between cryopreserved and fresh cells. Cryopreservation did not change the effect of known CYP3A4 inducers that have poor cell permeability, indicating that cryopreservation had little effect on membrane permeability. In addition, cryopreserved HepG2 cells did not exhibit enhanced susceptibility to cytotoxic compounds compared to transiently transfected control cells. The use of cryopreserved cells enables this assay to run with enhanced efficiency.

Salvinorin A ((2S,4aR,6aR,7R,9S,10aS, 10bR)-2H-naphtho[2,1-c]pyran-7-carboxylic acid, 9-(acetyloxy)-2-(3-furanyl)dodecahydro-6a,10b-dimethyl-4,10-dioxo methyl ester, 1, CAS 83729-01-5) has been shown to bind with high affinity and selectivity to the kappa-opioid receptor (KOR) as an agonist. Bioisosteres of 1 were developed and biologically evaluated in binding and functional assays. The C-2 thioacetate isoster produced comparable activity to 1, but nitrogen substitution had a diminishing effect. Intermediates, which lack a beta-carbonyl at C-2, displayed moderate affinity. The derivatives were tested against all opioid subtypes and were selective towards KOR.

Salvinorin A is a naturally occurring hallucinogenic diterpenoid from the plant Salvia divinorumthat selectively and potently activates kappa-opioid receptors (KORs). Salvinorin A is unique in that it is the only known lipid-like molecule that selectively and potently activates a G-protein coupled receptor (GPCR), which has as its endogenous agonist a peptide; salvinorin A is also the only known non-nitrogenous opioid receptor agonist. In this paper, we identify key residues in KORs responsible for the high binding affinity and agonist efficacy of salvinorin A. Surprisingly, we discovered that salvinorin A was stabilized in the binding pocket by interactions with tyrosine residues in helix 7 (Tyr313 and Tyr320) and helix 2 (Tyr119). Intriguingly, activation of KORs by salvinorin A required interactions with the helix 7 tyrosines Tyr312, Tyr313, and Tyr320 and with Tyr139 in helix 3. In contrast, the prototypical nitrogenous KOR agonist U69593 and the endogenous peptidergic agonist dynorphin A (1-13) showed differential requirements for these three residues for binding and activation. We also employed a novel approach, whereby we examined the effects of cysteine-substitution mutagenesis on the binding of salvinorin A and an analogue with a free sulfhydryl group, 2-thiosalvinorin B. We discovered that residues predicted to be in close proximity, especially Tyr313, to the free thiol of 2-thiosalvinorin B when mutated to Cys showed enhanced affinity for 2-thiosalvinorin B. When these findings are taken together, they imply that the diterpenoid salvinorin A utilizes unique residues within a commonly shared binding pocket to selectively activate KORs.

A simple and efficient synthesis of 3,5‐dibromo‐O‐methyl‐L‐tyrosine is described by direct bromination of commercial O‐methyl‐L‐tyrosine with bromine in aqueous hydrochloric acid. MO calculations are presented to explain a difference in the degree of substitution of O‐methyl‐L‐tyrosine in different solvents.

The diterpene salvinorin A from Salvia divinorum has recently been reported to be a high-affinity and selective kappa-opioid receptor agonist (Roth et al., 2002). Salvinorin A and selected derivatives were found to be potent and efficacious agonists in several measures of agonist activity using cloned human kappa-opioid receptors expressed in human embryonic kidney-293 cells. Thus, salvinorin A, salvinorinyl-2-propionate, and salvinorinyl-2-heptanoate were found to be either full (salvinorin A) or partial (2-propionate, 2-heptanoate) agonists for inhibition of forskolin-stimulated cAMP production. Additional studies of agonist potency and efficacy of salvinorin A, performed by cotransfecting either the chimeric G proteins Gaq-i5 or the universal G protein Ga16 and quantification of agonist-evoked intracellular calcium mobilization, affirmed that salvinorin A was a potent and effective kappa-opioid agonist. Results from structure-function studies suggested that the nature of the substituent at the 2-position of salvinorin A was critical for kappa-opioid receptor binding and activation. Because issues of receptor reserve complicate estimates of agonist efficacy and potency, we also examined the agonist actions of salvinorin A by measuring potassium conductance through G protein-gated K(+) channels coexpressed in Xenopus oocytes, a system in which receptor reserve is minimal. Salvinorin A was found to be a full agonist, being significantly more efficacious than (trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl] benzeneacetamide methane-sulfonate hydrate (U50488) or (trans)-3,4-dichloro-N-methyl-N-[2-(1-pyrrolidinyl)-cyclohexyl] benzeneacetamide methane-sulfonate hydrate (U69593) (two standard kappa-opioid agonists) and similar in efficacy to dynorphin A (the naturally occurring peptide ligand for kappa-opioid receptors). Salvinorin A thus represents the first known naturally occurring non-nitrogenous full agonist at kappa-opioid receptors.

This thesis draws on the work of a wide range of theorists, philosophers, and educators in order to examine corporate sponsored curriculum used in the Canadian public school system. The main aim of this thesis is to determine whether or not corporate sponsored curriculum is, in fact, advertising. Does the classroom become a marketing medium through the use of corporate sponsored curriculum? Two case studies, of the Bank of Montreal's my money Investment Kit and the Tampax Health Education Program On Puberty And Menstrual Health, will be used as models for this thesis. The curriculum will be examined through both proponents and critics of adverting via four main perspectives. These perspectives include: (1) the view that advertising is a form of information, (2) the view that advertising is a form of persuasion, (3) the view that advertising is a form of symbolism, and (4) the critical cultural view of advertising. The findings will be entrenched in the socio-historical context of the current role played by the system of public education. This role is then contrasted with the aims and desires of marketing organizations and corporations. The thesis concludes with a synthesis of the findings and combines them with theoretical, philosophical, and contemporary views of education and society. Source: Masters Abstracts International, Volume: 39-02, page: 0372. Adviser: Myles Ruggles. Thesis (M.A.)--University of Windsor (Canada), 2000.