Enantiomeric methadone quantitation on real post-mortem dried matrix spots samples: Comparison of liquid chromatography and supercritical fluid chromatography coupled to mass spectrometry.

This study describes two bioanalytical methods for the quantitation of the two methadone enantiomers in dried matrix spots using high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) and high performance supercritical chromatography tandem mass spectrometry (HPSFC-MS/MS). Dried matrix spots were obtained by spotting 10 µL of each sample fluid on a Whatman paper. Methadone and its main metabolite, EDDP, were extracted with 100 µL methanol and subsequently injected into the LC-MS/MS and SFC-MS/MS systems. Enantiomeric separation was achieved with AGP-column for the LC conditions and with Chiralpak IH-3 in SFC. The two methods were fully validated and 93 post-mortem samples were analysed with both analytical methods. Results from validation parameters and results obtained for all post-mortem samples were compared with a significant spearman correlation of rs = 0.9978 for R-methadone and rs = 0.9981 for S-methadone. The LC method provided better results in terms of uncertainty, retention factor and resolution, whereas SFC provides better sensitivity, with lower LOD. Median R-/S-methadone ratio in peripheral blood was found equal to 1.60 (N = 32), varying from 0.79 to 4.23. The reported values were in good agreement with previously published results. Based on the results obtained here, SFC-MS/MS can be considered a reliable alternative to the widely used LC-MS/MS for the quantitation of methadone enantiomers in bioanalysis and should be evaluated for other bioanalytical methods. Both methods can be easily and quickly used in toxicological routine analysis for the methadone quantitation in human fluids matrices, even if considering that the polysaccharide coated column IH-3 used in SFC does not allow the enantiomeric EDDP separation.

Isotonitazene: Fatal intoxication in three cases involving this unreported novel psychoactive substance in Switzerland.

The paper describes the first three deaths reported in Europe involved in isotonitazene consumption, a potent benzimidazole derivate opioid consumed in the recreational drug scene. Isotonitazene powder and purity determination was performed on the sample collected in the first death scene by NMR, HRMS, GC-FTIR, ATR-FTIR and GC-MS. Isotonitazene purity was determined by GC-MS analysis and proton NMR, and was defined to be above 95 % and 98 %, respectively. Quantification of isotonitazene in biological samples was performed using a targeted analysis based on SPE extraction and ultra-high performance liquid chromatography tandem mass spectrometry. The isotonitazene median concentration in femoral whole blood was 1.20ng/mL. Isotonitazene concentration in hair was similar or even lower compared to that seen in fentanyl abusers. Isotonitazene distribution in tissues converges in the brain, lungs and heart, respectively. Surprisingly, isotonitazene concentration in liver is the lowest measured for all tissues and fluids analyzed. Based on circumstantial evidence, autopsy findings and the results of the toxicological analysis, the medical examiner concluded that the cause of all three deaths was an acute intoxication with isotonitazene. Since isotonitazene toxic concentration levels are very low, the consumption of this new psychoactive drug is a real hazard for human health.

Tempo-spatial gait adaptations in stroke patients when approaching and crossing an elevated surface.

BACKGROUND: In ambulatory stroke survivors, outdoor walking is important for participation, so adapting to heightened levels (e.g. curbs) is essential. This needs precise step regulation and foot positioning and has to be achieved despite impaired balance and motor regulation. RESEARCH QUESTION: How do stroke patients approach and cross elevated surfaces? METHODS: Gait of 12 hemiparetic stroke patients (62.8 ± 10.3 years; Functional Ambulatory Category 3-5) and 13 controls (60.0 ± 12.4 years) was compared using a sensor carpet and 3D motion capturing to collect tempo-spatial parameters and foot trajectories in two conditions: flat walking vs. approaching to and stepping onto an elevated surface (height 15 cm) in a self-selected manner (6 trials each). Tempo-spatial adaptations were normalized to flat walking while trajectory analysis focused on foot clearance and placement. Complementary assessments included the Dynamic-Gait-Index, the Berg-Balance-Test and the Falls Efficacy Scale. RESULTS: Patients showed significantly worse Dynamic-Gait-Indices, less balance and more fear of falling. During the approach phase, patients slowed down, partly accompanied by shorter steps which controls did not. During crossing, no preference for a specific leading leg was detected. Clearance of the leading leg on average was not reduced but patients landed closer to the edge. Still clearance of the paretic leg was less than that of the non-paretic leg and the minimal clearance across all trials suggested an increased tripping risk, most evident for the trailing leg. In particular slower approaching caused difficulties to ensure sufficient leg clearance and to place the foot safely. Independent from that, better balance correlated with safer clearance. SIGNIFICANCE: When managing elevated levels, leading with the paretic leg causes more difficulties to safely clear the legs which is considerably dependent upon speed. Therapists should consider that slow walking may not increase safety while faster gait and aspects of postural control potentially facilitate crossing a curb.

[Off-label biological therapies in patients with large vessel vasculitis and/or polymyalgia rheumatica : Safety and efficacy analysis of a nationwide German registry (GRAID2)]. / Off-label-Biologikatherapie bei Patienten mit Großgefäßvaskulitiden und/oder Polymyalgia rheumatica : Wirksamkeits- und Sicherheitsanalyse eines nationalen Registers (GRAID2).

OBJECTIVE: To evaluate the safety and clinical outcome of biological therapies in patients with large vessel vasculitis (LVV) or polymyalgia rheumatica (PMR) refractory to standard of care therapy in a real-life setting in Germany. METHODS: GRAID 2 (German Registry in Autoimmune Diseases 2) is a retrospective, noninterventional, multicenter registry collecting data from all patients with inflammatory rheumatic diseases refractory to conventional therapy treated with an initial off-label biological between August 2006 and December 2013. The retrospective documentation comprised case history, diagnosis, course of disease including safety and overall efficacy. RESULTS: Data from 14 patients were collected, 11 with LVV (78.6%) and 3 with isolated PMR (21.4%). Ten patients were treated with tocilizumab (71.4%), while 3 patients received infliximab infusions (21.4%) and 1 patient was treated with rituximab (7.1%). All clinical as well as laboratory efficacy parameters improved substantially. After the first application, tolerability of biologicals was assessed as "very good"/"good" by the physicians in 92.3% of the patients. Altogether, 8 adverse events (AEs) occurred in 4 patients including 3 infections (1 urogenital infection, 2 diverticulitis) representing a rate of 23.6 infections per 100 patient-years. One of these infections (diverticulitis under infliximab treatment) was rated as serious AE, requiring ICU treatment representing a rate of serious AEs of 7.9 per 100 patient-years. No deaths occurred during the observation period. CONCLUSION: With known limitations of a retrospective database, the results of this survey confirm data of smaller case series and proof-of-concept studies and suggest a substantial response to biological therapies in patients with otherwise refractory LVV or PMR with no new safety signals.

Acute effects of LSD on amygdala activity during processing of fearful stimuli in healthy subjects.

Lysergic acid diethylamide (LSD) induces profound changes in various mental domains, including perception, self-awareness and emotions. We used functional magnetic resonance imaging (fMRI) to investigate the acute effects of LSD on the neural substrate of emotional processing in humans. Using a double-blind, randomised, cross-over study design, placebo or 100 µg LSD were orally administered to 20 healthy subjects before the fMRI scan, taking into account the subjective and pharmacological peak effects of LSD. The plasma levels of LSD were determined immediately before and after the scan. The study (including the a priori-defined study end point) was registered at ClinicalTrials.gov before study start (NCT02308969). The administration of LSD reduced reactivity of the left amygdala and the right medial prefrontal cortex relative to placebo during the presentation of fearful faces (P<0.05, family-wise error). Notably, there was a significant negative correlation between LSD-induced amygdala response to fearful stimuli and the LSD-induced subjective drug effects (P<0.05). These data suggest that acute administration of LSD modulates the engagement of brain regions that mediate emotional processing.

Evaluation of a Novel Semi-Automated Ultrasound System for the Detection of Synovitis: A Prospective Study involving 45 Patients with Rheumatoid Arthritis.

Background: Arthrosonography has proven to be more sensitive and reliable for the detection of synovitis than clinical examination, but a comprehensive examination of small joints is time-consuming. The automated breast volume scanner (ABVS) has been developed to allow automatic and reproducible series of consecutive B-mode pictures of the female breast. Objectives: To analyze the comparability of ABVS and conventional manual ultrasonography (mUS) for the detection of synovitis in hands and feet of patients with rheumatoid arthritis (RA). Methods: 45 patients with early and established active rheumatoid arthritis were recruited for this trial. All subjects were assessed clinically and by manual (Esaote MyLab70) and automated ultrasound (ACUSON S2000™ ABVS). The wrists, the metacarpophalangeal and proximal interphalangeal joints of the hands and the metatarsophalangeal joints of the feet were examined. Results: A total of 2 340 joint aspects were examined with both methods. ABVS detected 291 grade 1, 124 grade 2, 100 grade 3 cases of synovitis (515 in total) compared to 267, 180 and 145 cases of synovitis (592 in total) with mUS. 242 erosions and 52 cases of tenosynovitis were found by ABVS compared to 244 erosions and 99 cases of tenosynovitis found by mUS. Kappa coefficients for the agreement between both methods ranged from 0.51 in PIP joints to 0.71 in MCP joints. The correlations with clinical parameters as well as interrater agreements were comparable for both ultrasound methods. Conclusion: Based on the results, ABVS seems to be a promising technology for the comprehensive and time-saving assessment of synovitis in RA.

Subjective body vertical: a promising diagnostic tool in idiopathic normal pressure hydrocephalus?

Postural instability is a frequent symptom of patients with idiopathic normal pressure hydrocephalus (iNPH), and might be due to the misperception of body verticality. The objective of this study was to assess the usefulness of the subjective body vertical (SBV) as a potential tool for diagnosing iNPH. Twenty patients with iNPH underwent tests of SBV in the pitch and roll planes before and after cerebrospinal fluid (CSF) drainage. Ten patients with other central gait disorders served as controls and also underwent tests for SBV. Before CSF drainage, patients with iNPH showed an impaired verticality perception in the pitch plane with a significant backward deviation of the SBV as compared to the control group (iNPH: mean ± SD -3.7 ± 3.6°; control group: -0.8 ± 2.2°; t value = -2.30, p t-test = 0.03). After CSF drainage, the SBV of the iNPH patients normalized for the pitch plane (-0.9 ± 1.9°). There was a correlation between the backward deviation of the SBV and the ventricular enlargement of the frontal horns (Evan's index; r = -0.52; p Pearson = 0.02). An even stronger correlation was found with the enlargement of the third ventricle (Thalamus index; r = -0.64; p Pearson = 0.002). The new and clinically relevant finding of this study is that verticality perception of patients with iNPH is primarily impaired the pitch plane, and it improves after CSF drainage. This disturbance in pitch might be due to a bilateral central vestibular dysfunction of the thalamus. Determination of the SBV in pitch promises to increase diagnostic accuracy in the cases of suspected iNPH.

Neuroimaging in moderate MDMA use: A systematic review.

MDMA ("ecstasy") is widely used as a recreational drug, although there has been some debate about its neurotoxic effects in humans. However, most studies have investigated subjects with heavy use patterns, and the effects of transient MDMA use are unclear. In this review, we therefore focus on subjects with moderate use patterns, in order to assess the evidence for harmful effects. We searched for studies applying neuroimaging techniques in man. Studies were included if they provided at least one group with an average of <50 lifetime episodes of ecstasy use or an average lifetime consumption of <100 ecstasy tablets. All studies published before July 2015 were included. Of the 250 studies identified in the database search, 19 were included. There is no convincing evidence that moderate MDMA use is associated with structural or functional brain alterations in neuroimaging measures. The lack of significant results was associated with high methodological heterogeneity in terms of dosages and co-consumption of other drugs, low quality of studies and small sample sizes.

Ballistic Beloniformes attacking through Snell's Window.

Needlefishes (Beloniformes) were observed employing a range of stalking and attacking behaviours to attack schools of bait fishes ranging from the use of tactics common to predatory fishes to a novel behaviour: the use of leaping, aerial attacks. These aerial attacks are suggested to serve two purposes: to extend the attack range of the needlefishes and to reduce their prey's potential for evasion. Furthermore, a third purpose is hypothesized that the needlefishes are taking advantage of Snell's Window, an optical effect which may mask their approach to their prey.

An advanced lithium-air battery exploiting an ionic liquid-based electrolyte.

A novel lithium-oxygen battery exploiting PYR14TFSI-LiTFSI as ionic liquid-based electrolyte medium is reported. The Li/PYR14TFSI-LiTFSI/O2 battery was fully characterized by electrochemical impedance spectroscopy, capacity-limited cycling, field emission scanning electron microscopy, high-resolution transmission electron microscopy, and X-ray photoelectron spectroscopy. The results of this extensive study demonstrate that this new Li/O2 cell is characterized by a stable electrode-electrolyte interface and a highly reversible charge-discharge cycling behavior. Most remarkably, the charge process (oxygen oxidation reaction) is characterized by a very low overvoltage, enhancing the energy efficiency to 82%, thus, addressing one of the most critical issues preventing the practical application of lithium-oxygen batteries.

Translation of genomics-guided RNA-based personalised cancer vaccines: towards the bedside.

Cancer is a disease caused by DNA mutations. Cancer therapies targeting defined functional mutations have shown clinical benefit. However, as 95% of the mutations in a tumour are unique to that single patient and only a small number of mutations are shared between patients, the addressed medical need is modest. A rapidly determined patient-specific tumour mutation pattern combined with a flexible mutation-targeting drug platform could generate a mutation-targeting individualised therapy, which would benefit each single patient. Next-generation sequencing enables the rapid identification of somatic mutations in individual tumours (the mutanome). Immunoinformatics enables predictions of mutation immunogenicity. Mutation-targeting RNA-based vaccines can be rapidly and affordably synthesised as custom GMP drug products. Integration of these cutting-edge technologies into a clinically applicable process holds the promise of a disruptive innovation benefiting cancer patients. Here, we describe our translation of the individualised RNA-based cancer vaccine concept into clinic trials.

[Segmental testicular infarction in sickle cell anemia]. / Segmentaler Hodeninfarkt bei Sichelzellenanämie.

Vascular occlusions are the clinical indicators of sickle cell disease and in urology they can lead to papillary necrosis, renal infarction or priapism. Segmental testicular infarction in patients with sickle cell disease is a rare event and only a few cases have been reported. We present a 25-year-old man with right testicular pain increasing over 3 days and sickle cell disease. Ultrasound of the right scrotum presented an inhomogeneous, mainly hypoechegenic mass with a hyperechogenic margin and no sign of blood flow. A partial orchiectomy was performed with total enucleation of the lesion, which was histologically diagnosed as benign hemorrhagic necrotic testicular tissue.

Gender-specific elimination of continuous-infusional 5-fluorouracil in patients with gastrointestinal malignancies: results from a prospective population pharmacokinetic study.

BACKGROUND: This study was initiated to assess the quantitative impact of patient anthropometrics and dihydropyrimidine dehydrogenase (DPYD) mutations on the pharmacokinetics (PK) of 5-fluorouracil (5FU) and to explore limited sampling strategies of 5FU. PATIENTS AND METHODS: We included 32 patients with gastrointestinal malignancies, receiving 46-h continuous-infusional 5FU and performed PK-sampling at baseline, 15, 30, 45 min, 1 and 2 h after the start of infusion and at the end of infusion, for 2 subsequent cycles. Plasma concentrations of 5FU, 5-fluorodihydrouracil (5FUH2), uracil (U) and 5,6-dihydrouracil (UH2) were determined using LC-MS/MS and submitted to population PK analysis using nonlinear mixed-effects modeling. Broad genotyping of DPYD was performed, and the potential impact of the DPYD genotype on the elimination of 5FU was assessed. Limited sampling strategies were evaluated for their accuracy to predict steady-state concentrations of 5FU (CSS(5FU)), using data simulations based on the final PK-model. RESULTS: The area-under-the concentration-time curve of 5FU (AUC(5FU)) was found to be <20 mg h/L in 33 occasions (58 %), between 20 and 30 mg h/L in 17 occasions (30 %) and >30 mg h/L in 7 occasions (12 %). Men had a 26 % higher elimination of 5FU and a 18 % higher apparent elimination of 5FUH2. Accordingly, women had a higher AUC(5FU) compared to men (22 vs. 18 mg h/L, p = 0.04). No DPYD risk variants were found, and the DPYD variants detected (c.496A>G, c.1601G>A, c.1627A>G) were not significantly associated with the elimination of 5FU. Individual baseline UH(2)/U ratio was significantly associated with AUC(5FU) (R = -0.49, p < 0.001). Limited sampling strategies with time-points <3 h after the start of infusion were not adequate to predict CSS(5FU). Female gender was the only predictor of nausea/emesis in the multivariate model. CONCLUSIONS: Gender-specific elimination of 5FU is supported by the present data and may partly explain the gender-specific association between DPYD risk variants and 5FU-specific toxicity.

Molecular combination of the dopamine and serotonin scaffolds yield in novel antipsychotic drug candidates - characterization by in vivo experiments.

Serotonin and dopamine play an important role in the aetiology of schizophrenia. Combination of the structural scaffolds of both neurotransmitters in a single molecule lead to aromatic [d,g]-bisannelated azecine derivatives, which have been shown to be nanomolar to subnanomolar dopamine D1-D5 receptor antagonists with a preference for the D1 family. In this work the potential antipsychotic activity of some azecine derivatives was predicted by their dopamine receptor affinities obtained in vitro from radioligand binding experiments and conclusively confirmed in vivo (rats) by applying a conditioned avoidance model. Furthermore, the compounds were tested in vivo for the development of catalepsy, which is a predictive parameter for extra-pyramidal side-effects caused by many antipsychotics. The investigated azecines displayed low cytotoxicity, and the affinities for human dopamine D1-D5 and serotonin 5-HT2 A receptors were in a nanomolar range. In vivo, their antipsychotic activities in the rat model were comparable with those of haloperidol and risperidone, but revealed a 2-5 times better therapeutic range with regard to catalepsy. Preliminary tests for oral bioavailability also revealed promising results for this new class of potential antipsychotic compounds. In conclusion, our in vivo experiments show that aromatic [d,g]-annelated azecines represent a novel and advantageous class of potential atypical neuroleptics.

Outcome of intravenous thrombolysis in stroke patients weighing over 100 kg.

BACKGROUND: Intravenous thrombolysis with alteplase for ischemic stroke is fixed at a maximal dose of 90 mg for safety reasons. Little is known about the clinical outcomes of stroke patients weighing >100 kg, who may benefit less from thrombolysis due to this dose limitation. METHODS: Prospective data on 1,479 consecutive stroke patients treated with intravenous alteplase in six Swiss stroke units were analyzed. Presenting characteristics and the frequency of favorable outcomes, defined as a modified Rankin scale (mRS) score of 0 or 1, a good outcome (mRS score 0-2), mortality and symptomatic intracranial hemorrhage (SICH) were compared between patients weighing >100 kg and those weighing ≤100 kg. RESULTS: Compared to their counterparts (n = 1,384, mean body weight 73 kg), patients weighing >100 kg (n = 95, mean body weight 108 kg) were younger (61 vs. 67 years, p < 0.001), were more frequently males (83 vs. 60%, p < 0.001) and more frequently suffered from diabetes mellitus (30 vs. 13%, p < 0.001). As compared with patients weighing ≤100 kg, patients weighing >100 kg had similar rates of favorable outcomes (45 vs. 48%, p = 0.656), good outcomes (58 vs. 64%, p = 0.270) and mortality (17 vs. 12%, p = 0.196), and SICH risk (1 vs. 5%, p = 0.182). After multivariable adjustment, body weight >100 kg was strongly associated with mortality (p = 0.007) and poor outcome (p = 0.007). CONCLUSION: Our data do not suggest a reduced likehood of favorable outcomes in patients weighing >100 kg treated with the current dose regimen. The association of body weight >100 kg with mortality and poor outcome, however, demands further large-scale studies to replicate our findings and to explore the underlying mechanisms.

Femto-Newton light force measurement at the thermal noise limit.

The measurement of very small light forces has wide applications in many fields of physics. A common measurement method for small force detection is the determination of changes in the dynamic behavior of mechanical oscillators, either in amplitude or in frequency. The detection of slowly varying forces mostly requires long period oscillators, such as a torsion pendulum. We demonstrate the application of a macroscopic, low-noise, torsion balance oscillator for the detection of radiation pressure forces at the femto-Newton level. The system is "precooled" (removing excess seimic noise) to be only thermal noise limited. The demonstrated force sensitivity reaches the thermal limit.

Cost of injuries from a prospective cohort study of North Carolina high school athletes.

OBJECTIVE: To estimate the economic cost of injuries in a population of US high school varsity athletes. DESIGN AND SETTING: The North Carolina High School Athletic Injury Study, conducted from 1996 to 1999, was a prospective cohort study of injury incidence and severity. A two-stage cluster sampling technique was used to select athletic teams from 100 high schools in North Carolina. An injury cost model was used to estimate the economic cost of injury. PARTICIPANTS: Varsity athletes from 12 sports: football, girls' and boy's soccer, girls' and boys' track, girls' and boy's basketball, baseball, softball, wrestling, volleyball, and cheerleading. MAIN OUTCOME MEASURES: Descriptive data were collected at the time of injury. Three types of costs were estimated: medical, human capital (medical costs plus loss of future earnings), and comprehensive (human capital costs plus lost quality of life). RESULTS: The annual statewide estimates were $9.9 million in medical costs, $44.7 million in human capital costs, and $144.6 million in comprehensive costs. The mean medical cost was $709 per injury (95% CI $542 to $927), $2223 per injury (95% CI $1709 to $2893) in human capital costs, and $10,432 per injury (95% CI $8062 to $13,449) in comprehensive costs. Sport and competition division were significant predictors of injury costs. CONCLUSIONS: Injuries among high school athletes represent a significant economic cost to society. Further research should estimate costs in additional populations to begin to develop cost-effective sports injury prevention programs.

Comparative biology of human and canine osteosarcoma.

Dogs with osteosarcoma provide an important model for the same disease in humans. In this report, the comparative nature of human and canine osteosarcoma including incidence and risk factors, clinical presentation and diagnosis, genetic abnormalities, biologic behaviour and prognostic factors, as well as treatment options are reviewed.