Orbitrap technology for comprehensive metabolite-based liquid chromatographic–high resolution-tandem mass spectrometric urine drug screening – Exemplified for cardiovascular drugs

LC–high resolution (HR)-MS well established in proteomics has become more and more important in bioanalysis of small molecules over the last few years. Its high selectivity and specificity provide best prerequisites for its use in broad screening approaches. Therefore, Orbitrap technology was tested for developing a general metabolite-based LC–HR-MS/MS screening approach for urinalysis of drugs necessary in clinical and forensic toxicology. After simple urine precipitation, the drugs and their metabolites were separated within 10 min and detected by a Q-Exactive mass spectrometer in full scan with positive/negative switching, and subsequent data dependent acquisition (DDA) mode. Identification criteria were the presence of accurate precursor ions, isotopic patterns, five most intense fragment ions, and comparison with full HR-MS/MS library spectra. The current library contains over 1900 parent drugs and 1200 metabolites. The method was validated for typical drug representatives and metabolites concerning recovery, matrix effects, process efficiency, and limits showed acceptable results. The applicability was tested first for cardiovascular drugs, which should be screened for in poisoning cases and for medication adherence of hypertension patients. The novel LC–HR-MS/MS method allowed fast, simple, and robust urine screening, particularly for cardiovascular drugs showing the usefulness of Orbitrap technology for drug testing.     

Mechanical Behavior of Unidirectional Curaua Fiber and Glass Fiber Composites

Summary: This work intends to promote the use of natural fibers by comparing the behavior of isophthalic polyester matrix composites reinforced with unidirectional curaua fibers with that of unidirectional glass fiber composites. The composites were produced varying the reinforcement angle (0°, 15°, 30°, 45°, 60°, 75° and 90°) with the aim of studying the fiber orientation effect on composite strength. Composites were also made varying the fiber volume fraction (10%, 20%, 30%, 40% and 50%). The efficiency of an alkaline (5% NaOH) surface treatment of the curaua fiber was also evaluated. The unidirectional composites were characterized using tensile, flexural and short beam tests as per ASTM standards. The properties of a lamina reinforced with either glass or curaua fibers were also studied using theoretical micromechanical approach available in commercial software. The curaua fiber alkaline treatment produced higher tensile strength results compared with untreated fibers. The increase in reinforcement angle significantly decreased strength and modulus of the composites, as expected, and the glass fiber composites showed a more pronounced dependence with fiber orientation. Although the glass fiber laminas showed the best mechanical performance, the results obtained with the curaua fibers were considered similar for angles greater than 45°.     

Aging, spatial cues, and single- versus dual-task performance in competing speech perception

Older individuals often report difficulty coping in situations with multiple conversations in which they at times need to "tune out" the background speech and at other times seek to monitor competing messages. The present study was designed to simulate this type of interaction by examining the cost of requiring listeners to perform a secondary task in conjunction with understanding a target talker in the presence of competing speech. The ability of younger and older adults to understand a target utterance was measured with and without requiring the listener to also determine how many masking voices were presented time-reversed. Also of interest was how spatial separation affected the ability to perform these two tasks. Older adults demonstrated slightly reduced overall speech recognition and obtained less spatial release from masking, as compared to younger listeners. For both younger and older listeners, spatial separation increased the costs associated with performing both tasks together. The meaningfulness of the masker had a greater detrimental effect on speech understanding for older participants than for younger participants. However, the results suggest that the problems experienced by older adults in complex listening situations are not necessarily due to a deficit in the ability to switch and/or divide attention among talkers.     

TRPM5-expressing microvillous cells in the main olfactory epithelium

Background  

The main olfactory epithelium (MOE) in the nasal cavity detects a variety of air borne molecules that provide information regarding the presence of food, predators and other relevant social and environmental factors. Within the epithelium are ciliated sensory neurons, supporting cells, basal cells and microvillous cells, each of which is distinct in morphology and function. Arguably, the least understood, are the microvillous cells, a population of cells that are small in number and whose function is not known. We previously found that in a mouse strain in which the TRPM5 promoter drives expression of the green fluorescent protein (GFP), a population of ciliated olfactory sensory neurons (OSNs), as well as a population of cells displaying microvilli-like structures is labeled. Here we examined the morphology and immunocytochemical properties of these microvillous-like cells using immunocytochemical methods.     

The effect of masker interaural phase on the detection of a monaural signal

The masking-level difference (MLD) for a 500-Hz monaural pure-tone signal was examined as a function of the interaural phase shift of a 100-Hz-wide noise band centered on 500 Hz. Results indicated that the MLD decreased in magnitude as the interaural phase shift of the masker increased. In a second experiment, the 100-Hz-wide noise band was used as both the masker and the signal in order to examine the detection cues of interaural time difference and interaural level difference separately. Again, the interaural phase of the masker was varied, and an Sm signal was presented. Results indicated that the MLD decreased as a function of increasing masker interaural temporal difference for the time cue, but that the MLD did not change systematically for the level cue. The deterioration of binaural detection as a function of increasing masker interaural phase difference was not as great as that which has been reported in localization and lateralization experiments.     

An adeno-associated viral vector transduces the rat hypothalamus and amygdala more efficient than a lentiviral vector

Background  

This study compared the transduction efficiencies of an adeno-associated viral (AAV) vector, which was pseudotyped with an AAV1 capsid and encoded the green fluorescent protein (GFP), with a lentiviral (LV) vector, which was pseudotyped with a VSV-G envelop and encoded the discosoma red fluorescent protein (dsRed), to investigate which viral vector transduced the lateral hypothalamus or the amygdala more efficiently. The LV-dsRed and AAV1-GFP vector were mixed and injected into the lateral hypothalamus or into the amygdala of adult rats. The titers that were injected were 1 × 10⁸ or 1 × 10⁹ genomic copies of AAV1-GFP and 1 × 10⁵ transducing units of LV-dsRed.     

The effect of topical anesthesia on vocal fold motion

The objective of this study was to determine if topical anesthesia to the larynx and pharynx affects vocal fold motion during dynamic voice evaluation with transnasal flexible endoscopy. Transnasal dynamic laryngeal examinations of 10 patients with no voice complaints were evaluated by five blinded fellowship-trained laryngologists. Each patient was examined before and after application of topical anesthetic. Reviewers rated briskness of right and left vocal fold movement and longitudinal tension on a visual analogue scale. Statistical comparisons were made between individual subject scores before and after anesthetic application. Inter-rater reliability was also assessed. No statistical difference was observed between subject scores before and after anesthetic application. Average intraclass correlation coefficients were 0.643 and 0.591 for pre- and postanesthesia scores, respectively. Application of topical anesthesia to the larynx and pharynx does not affect vocal fold motion.     

The angular momentum of gravitational radiation

Using the approach initiated by Penrose, we calculate the flux of angular momentum emitted by a gravitational system. In appropriate circumstances, we recover the formula Ashtekar and Streubel derived for the total emitted angular momentum.