A User-Friendly Sample Preparation Alternative for Manual and Automated LC-MS/MS Quantification of Testosterone.

We describe an LC-MS/MS method for serum testosterone using a novel extraction media, AC Extraction Plate™ (AC Plate,Tecan Schweiz). The AC Plate principle is essentially that of a liquid-liquid extraction (LLE) but employs a stationary nonpolar phase coated on the wells of 96-well plates instead of a nonmiscible organic solvent for partitioning testosterone out of serum, leaving interfering substances behind. This low complexity sample preparation protocol has been validated for and used in production in our laboratory with both manual and automated liquid handling.

Validation of a liquid chromatography-tandem mass spectrometry method for analyzing cannabinoids in oral fluid.

A liquid chromatography tandem mass spectrometry method was developed for quantifying ten cannabinoids in oral fluid (OF). This method utilizes OF collected by the Quantisal™ device and concurrently quantifies cannabinol (CBN), cannabidiol (CBD), Δ-tetrahydrocannabinol (THC), 11-hydroxy-Δ-THC (11-OH-THC), 11-nor-9-carboxy-Δ-THC (THC-COOH), 11-nor-9-carboxy-Δ-THC glucuronide (THC-COOH-gluc), Δ-THC glucuronide (THC-gluc), cannabigerol (CBG), tetrahydrocannabiverin (THCV), and Δ-tetrahydrocannabinolic acid A (THCA-A). Solid phase extraction was optimized using Oasis Prime HLB 30 mg 96-well plates.

Reproducibility assessment for a broad spectrum drug screening method from urine using liquid chromatography time-of-flight mass spectrometry.

During the reproducibility validation for a time-of-flight (TOF) high-resolution mass spectrometry (HRMS) method set up to detect 61 drugs of abuse commonly encountered in the toxicology laboratory, it was noticed that, a number of compounds were not identified correctly during the between run analysis; the most difficult compounds to identify were norpropoxyphene, morphine, norbuprenorphine, nortriptyline, EDDP and tramadol. In subsequent patient comparison studies, screening a panel of 338 analytes, the TOF-HRMS method correctly identified 211 analytes over two runs, but did not identify 127. A total of 11 false positive results were identified by manual review of the data to be the result of confirmation ion signal-to-noise ratio(s) < 3, although one false positive that was difficult to resolve (i.

Broad-Spectrum Drug Screening Using Liquid Chromatography-Hybrid Triple-Quadrupole Linear Ion Trap Mass Spectrometry.

Urine is processed with a simple C18 solid-phase extraction (SPE) and reconstituted in mobile phase. The liquid chromatography system (LC) injects 10 μL of extracted sample onto a reverse-phase LC column for gradient analysis with ammonium formate/acetonitrile mobile phases. Drugs in the column eluent become charged in the ion source using positive electrospray ionization (ESI).

High-Throughput Serum 25-Hydroxy Vitamin D Testing with Automated Sample Preparation.

Serum from bar-coded tubes, and then internal standard, are pipetted to 96-well plates with an 8-channel automated liquid handler (ALH). The first precipitation reagent (methanol:ZnSO4) is added and mixed with the 8-channel ALH. A second protein precipitating agent, 1 % formic acid in acetonitrile, is added and mixed with a 96-channel ALH.

Transmission advantage favors selfing allele in experimental populations of self-incompatible Witheringia solanacea (solanaceae).

The evolution of self-fertilization is one of the most commonly traversed transitions in flowering plants, with profound implications for population genetic structure and evolutionary potential. We investigated factors influencing this transition using Witheringia solanacea, a predominantly self-incompatible (SI) species within which self-compatible (SC) genotypes have been identified. We showed that self-compatibility in this species segregates with variation at the S-locus as inherited by plants in F1 and F2 generations.

On a wing and a prayer: an assessment of modularized crew resource management training for health care professionals.

Introduction: Evidence suggests that Crew Resource Management (CRM), a form of team training, is beneficial. In CRM training, participants learn individual portable team skills such as communication and decision making through group discussion and activities. However, the usual 1-day course format is not always compatible with health care organizational routines.

Embryonic inbreeding depression varies among populations and by mating system in Witheringia solanacea (Solanaceae).

Premise Of The Study: Embryonic inbreeding depression is a key influence on mating system evolution and can be difficult to estimate in self-incompatible species. A pollen chase experiment was used to estimate the magnitude of embryonic inbreeding depression in Costa Rican Witheringia solanacea, a species polymorphic for self-incompatibility (SI). In a pollen chase experiment, bud self-pollinations are followed after anthesis by outcross pollinations, with a comparable pair of outcross pollinations used as a control.

Moving interprofessional learning forward through formal assessment.

Context: There is increasing agreement that graduates who finish tertiary education with the full complement of skills and knowledge required for their designated profession are not 'work-ready' unless they also acquire interpersonal, collaborative practice and team-working capabilities. Health workers are unable to contribute to organisational culture in a positive way unless they too attain these capabilities. These capabilities have been shown to improve health care in terms of patient safety, worker satisfaction and health service efficiency.

An Interprofessional Collaborative Practice model for preparation of clinical educators.

Unlabelled: Work-integrated learning is essential to health professional education, but faces increasing academic and industry resource pressures. The aim of this pilot "Professional Practice Project" was to develop and implement an innovative education intervention for clinical educators across several health disciplines. The project used interprofessional collaboration as its underlying philosophy, and a participatory action research methodology in four cycles: Cycle 1: Formation of an interprofessional project executive and working party from academic staff.

Broad spectrum drug screening using liquid chromatography-hybrid triple quadrupole linear ion trap mass spectrometry.

Centrifuged urine, internal standard (promazine), and ammonium formate buffer are mixed in an autosampler vial to achieve a 10-fold dilution of the specimen. Without additional pretreatment, 10 microL of the sample is injected onto a C18 reverse phase column for gradient analysis with ammonium formate/acetonitrile mobile phases. Drugs in the column eluent become charged in the ion source using positive electrospray atmospheric pressure ionization.

Broad spectrum drug screening using electron-ionization gas chromatography-mass spectrometry (EI-GCMS).

A liquid-liquid extraction (LLE) of drugs and internal standard (promazine) is performed by mixing urine at basic pH with 1-chlorobutane. There are no hydrolysis or derivatization steps. After centrifugation the organic (upper) layer is transferred to another tube and evaporated.

Morbidity involving the hallucinogenic designer amines MDA and 2C-I.

A case is presented of a 39-year-old woman who suffered severe debilitation because of a hemorrhagic stroke in the context of substance abuse. The patient presented to the emergency room with rapidly diminishing mental status, hypertension, and vasoconstriction; her friends provided a history of ingestion of cocaine, 3,4-methylenedioxymethamphetamine (MDMA), and 2C-I, a novel designer amine. A multi-targeted LC-MS/MS method for sympathomimetic amines and related drugs in urine detected and quantified 2C-I and MDA, while ruling out MDMA.

Clinical utility of an LC-MS/MS seizure panel for common drugs involved in drug-induced seizures.

Background: Approximately 6% of new-onset seizures are drug-related, but there is currently no reliable way to determine if a seizure is drug-induced. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is a powerful tool that allows simultaneous detection of numerous analytes of diverse chemical nature in patient samples. This allows a single analysis to incorporate many compounds relevant to a particular clinical presentation, such as suspected drug-induced seizures.

A hydrophilic-interaction chromatography tandem mass spectrometry method for quantitation of serum s-adenosylmethionine in patients infected with human immunodeficiency virus.

Background: s-Adenosylmethionine (SAM)1 has been suggested as a diagnostic test and surrogate marker for Pneumocystis jirovecii pneumonia (PCP) in HIV-positive patients. In this study, we report a robust hydrophilic-interaction liquid chromatography tandem mass spectrometry (LC-MS/MS) assay that can be used to quantitate serum SAM in clinical laboratories.

Methods: Proteins in serum samples were precipitated using trichloroacetic acid.

An action research protocol to strengthen system-wide inter-professional learning and practice [LP0775514].

Background: Inter-professional learning (IPL) and inter-professional practice (IPP) are thought to be critical determinants of effective care, improved quality and safety and enhanced provider morale, yet few empirical studies have demonstrated this. Whole-of-system research is even less prevalent. We aim to provide a four year, multi-method, multi-collaborator action research program of IPL and IPP in defined, bounded health and education systems located in the Australian Capital Territory (ACT).

Variation in the self-incompatibility response within and among populations of the tropical shrub Witheringia solanacea (Solanaceae).

Breakdown of genetically enforced self-incompatibility (SI), an extremely common and important evolutionary transition in plants, has conventionally been conceived as a qualitative rather than a quantitative change. We evaluated qualitative and quantitative variation in SI for four populations of Witheringia solanacea in Costa Rica, examining growth of self-pollen tubes in pollinations of buds and mature flowers. We also measured levels of RNase production in styles to determine whether enzyme production was correlated with differences in self-rejection.

Efficacy and safety of tifacogin (recombinant tissue factor pathway inhibitor) in severe sepsis: a randomized controlled trial.

Context: The expression and release of tissue factor is a major trigger for the activation of coagulation in patients with sepsis. Tissue factor pathway inhibitor (TFPI) forms a complex with tissue factor and blood protease factors leading to inhibition of thrombin generation and fibrin formation.

Objectives: To determine if administration of tifacogin (recombinant TFPI) provides mortality benefit in patients with severe sepsis and elevated international normalized ratio (INR) and to assess tifacogin safety in severe sepsis, including patients with low INR.

Anther-stigma separation is associated with inbreeding depression in Datura stramonium, a predominantly self-fertilizing annual.

Genetically based variation in outcrossing rate generates lineages within populations that differ in their history of inbreeding. According to some models, mating-system modifiers in such populations will demonstrate both linkage and identity disequilibrium with fitness loci, resulting in lineage-specific inbreeding depression. Other models assert that differences among families in levels of inbreeding depression are mainly attributable to random accumulation of genetic load, unrelated to variation at mating-system loci.

Components of pollination effectiveness in Psychotria suerrensis, a tropical distylous shrub.

In this paper I report components of effectiveness for pollinators of a tropical distylous shrub, Psychotria suerrensis (Rubiaceae), which is visited by a variety of bees, wasps, and butterflies, and by two species of hummingbirds. In the field, I measured the following components of effectiveness: frequency of visits, evenness of visits across plants, and diurnal pattern of visits. I also used flight-cage experiments to compare pollentransfer abilities of euglossine bees and heliconiid butterflies.

THE EVOLUTION OF DISTYLY: POLLEN TRANSFER IN ARTIFICIAL FLOWERS.

A recent model by Lloyd and Webb derives conditions necessary for the evolution of distyly based on pollen-transfer probabilities between ancestral morphs and invading mutants. We used bumblebees visiting artificial flowers to measure the parameters of the model. Our findings supported the first evolutionary step proposed by the model, establishment of a stigma-height polymorphism.