Development and validation of a presumptive colour spot test method for the detection of piperazine analogues in seized illicit materials

The increasingly large quantities of potentially illicit samples received for confirmatory analysis highlights the importance and demand for preliminary testing procedures that are simple, rapid, selective, inexpensive and able to be used in the field. Colour testing fulfils the aforementioned requirements and is a technique frequently employed to achieve presumptive identification. Piperazine analogues (often marketed as 'legal ecstasy') are a group of psychoactive substances that have recently become established on the illicit drug market and are not effectively discriminated or identified by current colour testing methods. Herein, we report on the development and validation of a chemical spot test for piperazine analogues present in unknown seized materials using the spectrophotometric reagent, sodium 1,2-naphthoquinone-4-sulphonate (NQS). Primary testing revealed that NQS reacts almost instantly to form an intense, bright orange-red coloured complex with the representative piperazine 1-benzylpiperazine (BZP) at room temperature. The results of the test, assessed by colour development, were evaluated visually and variables affecting the coloured reaction were optimised. The colour test method was validated to meet requirements for use in preliminary screening, providing qualitative and reliable presumptive test results. Validation studies show that the characteristic colour change is unique to the piperazine class at room temperature, and is unaffected by the presence of common cutting agents, i.e. glucose and caffeine, in test samples of 5% purity, and other drugs such as N-methyl-3,4-methylenedioxyamphetamine (MDMA). The NQS reagent stability was found to be limited to storage in a refrigerated environment for no more than one week before results were affected. The operational limit of detection was found to be 40 μg. © 2013 The Royal Society of Chemistry

Reducing the risk of iatrogenic Creutzfeldt–Jakob disease by improving the cleaning of neurosurgical instruments

Background: In all, there have been 178 variant Creutzfeldt–Jakob disease (vCJD) patients diagnosed in the UK, with an estimated maximum 1:2000 carriage rate based on archived appendix and tonsil tissue, implying that infection may be rare but carriage relatively frequent. Previous workers have identified that maintenance of surgical instruments in a humid atmosphere after use and prior to cleaning assists cleaning efficacy. Recently the Department of Health/Advisory Committee on Dangerous Pathogens UK have recommended a surgical instrument cleanliness threshold post cleaning of <5 μg protein per instrument side. Aim: To quantify cleanliness of neurosurgical instruments and to investigate cost-effective measures for improved cleaning. Methods: Two instrument protein quantification methods were used: one based on the International Standard (15883 series) using sodium dodecyl sulphate elution and ortho-phthalaldehyde reaction, and a second in-situ protein fluorescence detection system (ProReveal) providing results per instrument side. In-vitro investigation of the efficacy of some commercial and in-house pre-clean wetting agents was undertaken using artificial test soil and stainless steel discs under standard conditions. In-vivo evaluation of best-performing in-vitro agents was undertaken on craniotomy sets. Findings: ProReveal technology demonstrated that 163 out of 187 (87%) neurosurgical instruments had <5 μg residual protein per instrument side. The use of proprietary National Health Service plastic bags and sterile water-soaked wound pads were equivalent in efficacy to commercial pre-cleaning wetting products and significantly less expensive. Conclusion: Although we demonstrate low in-situ protein levels on neurosurgical instruments and the beneficial effects of keeping instruments moist, other cleaning critical-control points such as instrument loading patterns should also be monitored

Investigating the impact of remote neuroanatomy education during the COVID-19 pandemic using online examination performance in a National Undergraduate Neuroanatomy Competition

Neuroanatomy is a notoriously challenging subject for medical students to learn. Due to the coronavirus disease-19 (COVID-19) pandemic, anatomical education transitioned to an online format. We assessed student performance in, and attitudes toward, an online neuroanatomy assessment compared to an in-person equivalent, as a marker of the efficacy of remote neuroanatomy education. Participants in the National Undergraduate Neuroanatomy Competition (NUNC) 2021 undertook two online examinations: a neuroanatomically themed multiple-choice question paper and anatomy spotter. Students completed pre- and post-examination questionnaires to gauge their attitudes toward the online competition and prior experience of online anatomical teaching/assessment. To evaluate performance, we compared scores of students who sat the online (2021) and in-person (2017) examinations, using 12 identical neuroradiology questions present in both years. Forty-six percent of NUNC 2021 participants had taken an online anatomy examination in the previous 12?months, but this did not impact examination performance significantly (p?>?0.05). There was no significant difference in examination scores between in-person and online examinations using the 12 neuroradiology questions (p?=?0.69). Fifty percent of participants found the online format less enjoyable, with 63% citing significantly fewer networking opportunities. The online competition was less stressful for 55% of participants. This study provides some evidence to suggest that student performance is not affected when undertaking online examinations and proposes that online neuroanatomy teaching methods, particularly for neuroradiology, may be equally as effective as in-person approaches within this context. Participants perceived online examinations as less stressful but raised concerns surrounding the networking potential and enjoyment of online events.Peer reviewe

The VLT-FLAMES Tarantula Survey III: A very massive star in apparent isolation from the massive cluster R136

VFTS 682 is located in an active star-forming region, at a projected distance of 29 pc from the young massive cluster R136 in the Tarantula Nebula of the Large Magellanic Cloud. It was previously reported as a candidate young stellar object, and more recently spectroscopically revealed as a hydrogen-rich Wolf-Rayet (WN5h) star. Our aim is to obtain the stellar properties, such as its intrinsic luminosity, and to investigate the origin of VFTS 682. To this purpose, we model optical spectra from the VLT-FLAMES Tarantula Survey with the non-LTE stellar atmosphere code CMFGEN, as well as the spectral energy distribution from complementary optical and infrared photometry. We find the extinction properties to be highly peculiar (RV ~4.7), and obtain a surprisingly high luminosity log(L/Lsun) = 6.5 \pm 0.2, corresponding to a present-day mass of ~150Msun. The high effective temperature of 52.2 \pm 2.5kK might be explained by chemically homogeneous evolution - suggested to be the key process in the path towards long gamma-ray bursts. Lightcurves of the object show variability at the 10% level on a timescale of years. Such changes are unprecedented for classical Wolf-Rayet stars, and are more reminiscent of Luminous Blue Variables. Finally, we discuss two possibilities for the origin of VFTS 682: (i) the star either formed in situ, which would have profound implications for the formation mechanism of massive stars, or (ii) VFTS 682 is a slow runaway star that originated from the dense cluster R136, which would make it the most massive runaway known to date.Comment: 5 pages, 5 figures, accepted by A&A Letter

Resistance exercise initiates mechanistic target of rapamycin (mTOR) translocation and protein complex co-localisation in human skeletal muscle

This is the final version. Available from the publisher via the DOI in this record.The mechanistic target of rapamycin (mTOR) is a central mediator of protein synthesis in skeletal muscle. We utilized immunofluorescence approaches to study mTOR cellular distribution and protein-protein co-localisation in human skeletal muscle in the basal state as well as immediately, 1 and 3 h after an acute bout of resistance exercise in a fed (FED; 20 g Protein/40 g carbohydrate/1 g fat) or energy-free control (CON) state. mTOR and the lysosomal protein LAMP2 were highly co-localised in basal samples. Resistance exercise resulted in rapid translocation of mTOR/LAMP2 towards the cell membrane. Concurrently, resistance exercise led to the dissociation of TSC2 from Rheb and increased in the co-localisation of mTOR and Rheb post exercise in both FED and CON. In addition, mTOR co-localised with Eukaryotic translation initiation factor 3 subunit F (eIF3F) at the cell membrane post-exercise in both groups, with the response significantly greater at 1 h of recovery in the FED compared to CON. Collectively our data demonstrate that cellular trafficking of mTOR occurs in human muscle in response to an anabolic stimulus, events that appear to be primarily influenced by muscle contraction. The translocation and association of mTOR with positive regulators (i.e. Rheb and eIF3F) is consistent with an enhanced mRNA translational capacity after resistance exercise.Biotechnology and Biological Science Research Council (BBSRC)Natural Sciences and Engineering Research Council (NSERC)China Scholarship CouncilNational Institute of Arthritis and Musculoskeletal and Skin DiseasesDepartment of Defens

Children and adolescents with all forms of shoulder instability demonstrate differences in their movement and muscle activity patterns when compared to age- and sex-matched controls.

BACKGROUND: Shoulder instability is a complex impairment and identifying biomarkers which differentiate subgroups is challenging. There is limited fundamental movement and muscle activity data for identifying different mechanisms for shoulder instability in children and adolescents which may inform subgrouping and treatment allocation. HYPOTHESIS: Children and adolescents with shoulder instability (irrespective of etiology) have differences in their movement and muscle activity profiles compared to age- and sex-matched controls (two-tailed). METHODS: Young people between eight to 18 years were recruited into two groups of shoulder instability (SI) or and age- and sex-matched controls (CG). All forms of SI were included and young people with co-existing neurological pathologies or deficits were excluded. Participants attended a single session and carried out four unweighted and three weighted tasks in which their movements and muscle activity was measured using 3D-movement analysis and surface electromyography. Statistical parametric mapping was used to identify between group differences. RESULTS: Data was collected for 30 young people (15 SI (6M:9F) and 15 CG (8M:7F)). The mean (SD) age for all participants was 13.6 years (3.0). The SI group demonstrated consistently more protracted and elevated sternoclavicular joint positions during all movements. Normalized muscle activity in Latissimus dorsi was lower in the SI group and had the most statistically significant differences across all movements. Where differences were identified, the SI group also had increased normalized activity of their middle trapezius, posterior deltoid and biceps muscles whilst activity of their latissimus dorsi, triceps and anterior deltoid were decreased compared to the CG group. No statistically significant differences were found for pectoralis major across any movements. Weighted tasks produced fewer differences in muscle activity patterns compared to unweighted tasks. DISCUSSION: Young people with SI may adapt their movements to minimize glenohumeral joint instability. This was demonstrated by reduced variability in acromioclavicular and sternoclavicular joint angles, adoption of different movement strategies across the same joints and increased activity of the scapular stabilizing muscles, despite achieving similar arm positions to the CG. CONCLUSION: Young people with shoulder instability demonstrated consistent differences in their muscle activity and movement patterns. Consistently observed differences at the shoulder girdle included increased sternoclavicular protraction and elevation accompanied by increased normalized activity of the posterior scapula stabilizing muscles. Existing methods of measurement may be used to inform clinical decision making, however, further work is needed evaluate the prognostic and clinical utility of derived 3D and sEMG data for informing decision making within shoulder instability

Is there a compact companion orbiting the late O-type binary star HD 164816?

We present a multi-wavelength (X-ray, $\gamma$-ray, optical and radio) study of HD 194816, a late O-type X-ray detected spectroscopic binary. X-ray spectra are analyzed and the X-ray photon arrival times are checked for pulsation. In addition, newly obtained optical spectroscopic monitoring data on HD 164816 are presented. They are complemented by available radio data from several large scale surveys as well as the \emph{FERMI} $\gamma$-ray data from its \emph{Large Area Telescope}. We report the detection of a low energy excess in the X-ray spectrum that can be described by a simple absorbed blackbody model with a temperature of $\sim$ 50 eV as well as a 9.78 s pulsation of the X-ray source. The soft X-ray excess, the X-ray pulsation, and the kinematical age would all be consistent with a compact object like a neutron star as companion to HD 164816. The size of the soft X-ray excess emitting area is consistent with a circular region with a radius of about 7 km, typical for neutron stars, while the emission measure of the remaining harder emission is typical for late O-type single or binary stars. If HD 164816 includes a neutron star born in a supernova, this supernova should have been very recent and should have given the system a kick, which is consistent with the observation that the star HD 164816 has a significantly different radial velocity than the cluster mean. In addition we confirm the binarity of HD 164816 itself by obtaining an orbital period of 3.82 d, projected masses $m_1 {\rm sin}^{3} i$ = 2.355(69) M$_\odot$, $m_2 {\rm sin}^{3} i$ = 2.103(62) M$_\odot$ apparently seen at low inclination angle, determined from high-resolution optical spectra.Comment: Accepted for publication by MNRAS, 11 pages, 6 figures, 4 table

A catalogue of young runaway Hipparcos stars within 3kpc from the Sun

Traditionally runaway stars are O and B type stars with large peculiar velocities.We want to extend this definition to young stars (up to ~50 Myr) of any spectral type and identify those present in the Hipparcos catalogue applying different selection criteria such as peculiar space velocities or peculiar one-dimensional velocities. Runaway stars are important to study the evolution of multiple star systems or star clusters as well as to identify origins of neutron stars. We compile distances, proper motions, spectral types, luminosity classes, V magnitudes and B-V colours and utilise evolutionary models from different authors to obtain star ages and study a sample of 7663 young Hipparcos stars within 3 kpc from the Sun. Radial velocities are obtained from the literature. We investigate the distributions of the peculiar spatial velocity, the peculiar radial velocity as well as the peculiar tangential velocity and its one-dimensional components and obtain runaway star probabilities for each star in the sample. In addition, we look for stars that are situated outside any OB association or OB cluster and the Galactic plane as well as stars of which the velocity vector points away from the median velocity vector of neighbouring stars or the surrounding local OB association/ cluster although the absolute velocity might be small. We find a total of 2547 runaway star candidates (with a contamination of normal Population I stars of 20 per cent at most). Thus, after subtraction of those 20 per cent, the runaway frequency among young stars is about 27 per cent. We compile a catalogue of runaway stars which will be available via VizieR.Comment: 12 pages, 8 figures, 7 tables, accepted for publication in MNRAS old version replaced due to change of the title after journal proof-readin

The evolution of irradiance detection: melanopsin and the non-visual opsins

Circadian rhythms are endogenous 24 h cycles that persist in the absence of external time cues. These rhythms provide an internal representation of day length and optimize physiology and behaviour to the varying demands of the solar cycle. These clocks require daily adjustment to local time and the primary time cue (zeitgeber) used by most vertebrates is the daily change in the amount of environmental light (irradiance) at dawn and dusk, a process termed photoentrainment. Attempts to understand the photoreceptor mechanisms mediating non-image-forming responses to light, such as photoentrainment, have resulted in the discovery of a remarkable array of different photoreceptors and photopigment families, all of which appear to use a basic opsin/vitamin A-based photopigment biochemistry. In non-mammalian vertebrates, specialized photoreceptors are located within the pineal complex, deep brain and dermal melanophores. There is also strong evidence in fish and amphibians for the direct photic regulation of circadian clocks in multiple tissues. By contrast, mammals possess only ocular photoreceptors. However, in addition to the image-forming rods and cones of the retina, there exists a third photoreceptor system based on a subset of melanopsin-expressing photosensitive retinal ganglion cells (pRGCs). In this review, we discuss the range of vertebrate photoreceptors and their opsin photopigments, describe the melanopsin/pRGC system in some detail and then finally consider the molecular evolution and sensory ecology of these non-image-forming photoreceptor systems

Normal calcium-activated anion secretion in a mouse selectively lacking TMEM16A in intestinal epithelium

Calcium-activated anion secretion is expected to ameliorate cystic fibrosis, a genetic disease that carries an anion secretory defect in exocrine tissues. Human patients and animal models of the disease that present a mild intestinal phenotype have been postulated to bear a compensatory calcium-activated anion secretion in the intestine. TMEM16A is calcium-activated anion channel whose presence in the intestinal epithelium is contradictory. We aim to test the functional expression of TMEM16A using animal models with Cftr and/or Tmem16a intestinal silencing. Expression of TMEM16A was studied in a wild type and intestinal Tmem16a knockout mice by mRNA-seq, mass-spectrometry, q-PCR, Western blotting and immunolocalization. Calcium-activated anion secretion was recorded in the ileum and proximal colon of these animals including intestinal Cftr knockout and double mutants with dual Tmem16a and Cftr intestinal ablation. Mucus homeostasis was studied by immune-analysis of Mucin-2 (Muc2) and survival curves were recorded. Tmem16a transcript was found in intestine. Nevertheless, protein was barely detected in colon samples. Electrophysiological measurements demonstrated that the intestinal deletion of Tmem16a did not change calcium-activated anion secretion induced by carbachol or ATP in ileum and proximal colon. Muc2 architecture was not altered by Tmem16a silencing as was observed when Cftr was deleted from mouse intestine. Tmem16a silencing neither affected animal survival nor modified the lethality observed in the intestinal Cftr-null mouse. Our results demonstrate that TMEM16A function in the murine intestine is not related to electrogenic calcium-activated anion transport and does not affect mucus homeostasis and survival of animals