Prevalence and drug use correlates of inadvertent fentanyl exposure among individuals misusing drugs in seven U.S. states.

Fentanyl has emerged as the leading cause of fatal drug overdoses in the U.S. Individuals misusing drugs may not always be aware of exposure to fentanyl.To determine the prevalence of fentanyl use and extent of awareness of fentanyl exposure among a national sample of treatment-seeking individuals with opioid use disorder (n = 1098).Participants provided oral fluid and urine specimens, which were tested for drugs by liquid chromatography/tandem mass spectrometry. Participants also provided self-reports of fentanyl use.49.5% tested positive for fentanyl in oral fluid, urine, or both. Of those testing positive for fentanyl, 29.8% were unaware that they had been exposed to fentanyl. Participants testing positive for opioids methadone, and specifically 6-monoacetylmorphine (6-MAM), a unique metabolite of heroin, were significantly more likely to be unaware of fentanyl exposure than participants testing negative for these substances, with a similar trend for oxycodone and tramadol.These findings may be due to fentanyl's effect being difficult to distinguish from that of other opioids, whereas when other types of drugs are adulterated with fentanyl, the differences in effects are likely to be readily discernable. These results support the importance of expanded drug-checking services.

Point (S-to-G) Mutations in the W(S/G)GE Motif in Red/Green Cyanobacteriochrome GAF Domains Enhance Thermal Reversion Rates.

Cyanobacteriochromes (CBCRs) are photoreceptors consisting of single or tandem GAF (cGMP-phosphodiesterase/adenylate cyclase/FhlA) domains that bind bilin chromophores. Canonical red/green CBCR GAF domains are a well-characterized subgroup of the expanded red/green CBCR GAF domain family that binds phycocyanobilin (PCB) and converts between a thermally stable red-absorbing Pr state and a green-absorbing Pg state. The rate of thermal reversion from Pg to Pr varies widely among canonical red/green CBCR GAF domains, with half-lives ranging from days to seconds. Since the thermal reversion rate is an important parameter for the application of CBCR GAF domains as optogenetic tools, the molecular factors controlling the thermal reversion rate are of particular interest. Here, we report that point mutations in a well-conserved W(S/G)GE motif alter reversion rates in canonical red/green CBCR GAF domains in a predictable manner. Specifically, S-to-G mutations enhance thermal reversion rates, while the reverse, G-to-S mutations slow thermal reversion. Despite the distance (>10 Å) of the mutation site from the chromophore, molecular dynamics simulations and nuclear magnetic resonance (NMR) analyses suggest that the presence of a glycine residue allows the formation of a water bridge that alters the conformational dynamics of chromophore-interacting residues, leading to enhanced Pg to Pr thermal reversion.

Impact of patient motivation on compliance and outcomes for incontinence.

OBJECTIVES: Physiotherapeutic treatment of pelvic floor dysfunction in women has variable outcomes, and identification of the predictors of outcomes is important in developing service provision and treatment planning. This study explored whether patient motivation affects treatment compliance and outcomes of physiotherapeutic treatment in a cohort of patients with pelvic floor dysfunction. DESIGN, SETTING AND PARTICIPANTS: A prospective observational study of 141 adult female patients referred consecutively to the physiotherapy clinic of an urogynaecological outpatients unit at a UK hospital. INTERVENTION: A 6-month (one group session per month) pelvic floor muscle training programme, which included a home exercise programme. MAIN OUTCOME MEASURES: Modified Oxford Grading System and Queensland Pelvic Floor Questionnaire ratings of pelvic floor function were taken pre- and post-intervention, and the University of Rhode Island Change Assessment Scale assessed patient motivation to change pre-intervention. RESULTS: Motivation to change predicted attendance at the intervention sessions and patient-rated improvements in function. Patients with higher baseline motivation to change also reported greater improvements in pelvic symptoms, given the same pelvic floor muscle strength improvement. CONCLUSIONS: Patient motivation affects physiotherapeutic treatment adherence and outcomes, and should be considered as part of future assessment/screening procedures. CLINICAL TRIAL REGISTRATION: NCT02549157.

Structure-based design of a photoswitchable affibody scaffold.

Photo-control of affinity reagents offers a general approach for high-resolution spatiotemporal control of diverse molecular processes. In an effort to develop general design principles for a photo-controlled affinity reagent, we took a structure-based approach to the design of a photoswitchable Z-domain, among the simplest of affinity reagent scaffolds. A chimera, designated Z-PYP, of photoactive yellow protein (PYP) and the Z-domain, was designed based on the concept of mutually exclusive folding. NMR analysis indicated that, in the dark, the PYP domain of the chimera was folded, and the Z-domain was unfolded. Blue light caused loss of structure in PYP and a two- to sixfold change in the apparent affinity of Z-PYP for its target as determined using size exclusion chromatography, UV-Vis based assays, and enyzme-linked immunosorbent assay (ELISA). A thermodynamic model indicated that mutations to decrease Z-domain folding energy would alter target affinity without loss of switching. This prediction was confirmed experimentally with a double alanine mutant in helix 3 of the Z-domain of the chimera (Z-PYP-AA) showing >30-fold lower dark-state binding and no loss in switching. The effect of decreased dark-state binding affinity was tested in a two-hybrid transcriptional control format and enabled pronounced light/dark differences in yeast growth in vivo. Finally, the design was transferable to the αZ-Taq affibody enabling tunable light-dependent binding both in vitro and in vivo to the Z-Taq target. This system thus provides a framework for the focused development of light switchable affibodies for a range of targets.

Yeast Two-Hybrid Screening of Photoswitchable Protein-Protein Interaction Libraries.

Although widely used in the detection and characterization of protein-protein interactions, Y2H screening has been under-used for the engineering of new optogenetic tools or the improvement of existing tools. Here we explore the feasibility of using Y2H selection and screening to evaluate libraries of photoswitchable protein-protein interactions. We targeted the interaction between circularly permuted photoactive yellow protein (cPYP) and its binding partner binder of PYP dark-state (BoPD) by mutating a set of four surface residues of cPYP that contribute to the binding interface. A library of ~10,000 variants was expressed in yeast together with BoPD in a Y2H format. An initial selection for the cPYP/BoPD interaction was performed using a range of concentrations of the cPYP chromophore. As expected, the majority (>90% of cPYP variants) no longer bound to BoPD. Replica plating was then used to evaluate the photoswitchability of the surviving clones. Photoswitchable cPYP variants with BoPD affinities equal to, or higher than, native cPYP were recovered in addition to variants with altered photocycles and binders that interacted with BoPD as apo-proteins. Y2H results reflected protein-protein interaction affinity, expression, photoswitchability, and chromophore uptake, and correlated well with results obtained both in vitro and in mammalian cells. Thus, by systematic variation of selection parameters, Y2H screens can be effectively used to generate new optogenetic tools for controlling protein-protein interactions for use in diverse settings.

A missense variant remote from the active site impairs stability of human phosphoglucomutase 1.

Missense variants of human phosphoglucomutase 1 (PGM1) cause the inherited metabolic disease known as PGM1 deficiency. This condition is categorised as both a glycogen storage disease and a congenital disorder of glycosylation. Approximately 20 missense variants of PGM1 are linked to PGM1 deficiency, and biochemical studies have suggested that they fall into two general categories: those affecting the active site and catalytic efficiency, and those that appear to impair protein folding and/or stability. In this study, we characterise a novel variant of Arg422, a residue distal from the active site of PGM1 and the site of a previously identified disease-related variant (Arg422Trp). In prior studies, the R422W variant was found to produce insoluble protein in a recombinant expression system, precluding further in vitro characterisation. Here we investigate an alternative variant of this residue, Arg422Gln, which is amenable to experimental characterisation presumably due to its more conservative physicochemical substitution. Biochemical, crystallographic, and computational studies of R422Q establish that this variant causes only minor changes in catalytic efficiency and 3D structure, but is nonetheless dramatically reduced in stability. Unexpectedly, binding of a substrate analog is found to further destabilise the protein, in contrast to its stabilising effect on wild-type PGM1 and several other missense variants. This work establishes Arg422 as a lynchpin residue for the stability of PGM1 and supports the impairment of protein stability as a pathomechanism for variants that cause PGM1 deficiency. SYNOPSIS: Biochemical and structural studies of a missense variant far from the active site of human PGM1 identify a residue with a key role in enzyme stability.

Data on the effect of high-pressure torsion processing on secondary cast Al-10%Si- Cu piston alloy: Methods, microstructure and mechanical characterizations.

The dataset presented here shows the microstructure and mechanical properties of secondary (recycled) cast aluminum-silicon (Al-Si) piston alloys processed through severe plastic deformation technique, known as high-pressure torsion (HPT). The HPT processing was undertaken for 1/4, 1/2, 1 and 10 turns of the lower anvil (rotating at constant speed of 1rpm) while the upper anvil maintained at a normal pressure of 3.0 GPa. The data on microstructural evolution obtained at the central region and edge of the circular (disk) HPT sample were obtained using optical and scanning electron microscopy and these data are presented here. The data on the analysis of the particle shape, sizes and distribution from the micrographs using ImageJ software are also presented. Data on mechanical properties characterized using Vickers microhardness measurement across the surface of HPT sample are also shown. Pictures depicting the microhardness measurement scheme, high-pressure torsion facility and sample nomenclature are presented.

Cystoscopic transurethral incision in simplex and duplex ureteroceles-is it the definitive procedure?

OBJECTIVE: The aim of this study is to evaluate the efficacy of primary cystoscopic transurethral incision (CTUI) in the management of paediatric ureteroceles. The secondary aim is to compare the efficacy of CTUI between simplex and duplex systems. PATIENTS AND METHODS: This is a retrospective review of consecutive paediatric patients requiring surgical intervention for ureterocele. Data collected for analysis included demographics, diagnostic, pre-operative investigations, operative interventions and postoperative variables. RESULTS: Over a 19-year period, 79 consecutive cases were identified, and 42 were male (53.2%). The mean follow-up was 6.7 years. Seventy-three (92.4%) cases underwent primary CTUI; 50 of these cases (68.5%) required no further procedures during the study period. Sixty-one cases were treated by endoscopic intervention alone (77.2%). Forty-one (51.9%) cases had a simplex system, and 38 (48.1%), a duplex system. There was no statistically significant difference in the efficacy of primary CTUI in simplex vs duplex systems. Of the 20 patients who had pre-operative and postoperative micturating cystourethrograms performed, seven (35%) developed de-novo postoperative vesicoureteric reflux after CTUI. CONCLUSION: Primary CTUI is a safe, minimally invasive procedure that is definitive in the majority of children presenting with a ureterocele that requires intervention. There was no difference in success of primary CTUI between simplex or duplex systems.

Enhanced Electrochemical Stability of TiO2-Protected, Al-doped ZnO Transparent Conducting Oxide Synthesized by Atomic Layer Deposition.

Transparent, conductive coatings on porous, three-dimensional materials are often used as the current collector for photoelectrode designs in photoelectrochemical applications. These structures allow for improved light trapping and absorption in chemically synthesized, photoactive overlayers while minimizing parasitic absorption in the current collecting layer. Atomic layer deposition (ALD) is particularly useful for fabricating transparent conducting oxides (TCOs) like Sn-doped In2O3 (ITO) and Al-doped ZnO (AZO) for structured materials because the deposition is specific to exposed surfaces. Unlike line-of-sight deposition methods (evaporation, spray pyrolysis, sputtering), ALD can access the entire complex interface to make a conformal transparent conductive layer. While ITO and AZO can be grown by ALD, they are intrinsically soluble in the acidic and basic environments common for electrochemical applications like water splitting. To take advantage of the unique characteristics of ALD in these applications, it is important to develop strategies for fabricating TCO layers with enhanced chemical stability. Ultrathin coatings of stable materials can be used to protect otherwise unstable electrochemical interfaces while maintaining the desired function. Here, we describe experiments to characterize the chemical and electrochemical stability of ALD-deposited AZO TCO thin films protected by a 10 nm TiO2 overlayer. The addition of a TiO2 protection layer is demonstrated to improve the chemical stability of AZO by orders of magnitude compared to unprotected, yet otherwise identically prepared, AZO films. The electrochemical stability is enhanced accordingly in both acidic and basic environments. We demonstrate that TiO2-protected AZO can be used as a TCO for both the cathodic hydrogen evolution (HER) and anodic water oxidation (OER) half-reactions of electrochemical water splitting in base and for HER in acid when the appropriate electrocatalysts are added. As a result, we show that ALD can be used to synthesize a chemically stable TCO heterostructure, expanding the range of materials and electrochemical environments available for building complex photoelectrode architectures.

A diffusion-based approach for modelling crack tip behaviour under fatigue-oxidation conditions.

Modelling of crack tip behaviour was carried out for a nickel-based superalloy subjected to high temperature fatigue in a vacuum and air. In a vacuum, crack growth was entirely due to mechanical deformation and thus it was sufficient to use accumulated plastic strain as a criterion. To study the strong effect of oxidation in air, a diffusion-based approach was applied to investigate the full interaction between fatigue and oxygen penetration at a crack tip. Penetration of oxygen into the crack tip induced a local compressive stress due to dilatation effect. An increase in stress intensity factor range or dwell times imposed at peak loads resulted in enhanced accumulation of oxygen at the crack tip. A crack growth criterion based on accumulated levels of oxygen and plastic strain at the crack tip was subsequently developed to predict the crack growth rate under fatigue-oxidation conditions. The predicted crack-growth behaviour compared well with experimental results.

Children with autism spectrum disorder: teaching conversation involving feelings about events.

BACKGROUND: Two procedures were developed to teach individuals with Autism Spectrum Disorders labels (tacts) for various private events (emotions): Study 1 attempted to distinguish them from pure tacts and mands (requests); and Study 2 attempted to train initiating a conversation with grammatically correct subject-verb-comment construction. METHODS: A multiple treatment reversal design was used in both studies, followed by a probe to see if the tacts were used across novel settings. RESULTS: The children were prompted to initiate a series of language exchanges, which resulted in an increased ability to participate in conversations about private events. CONCLUSIONS: Together, the results of both studies suggest that, by providing an effective and reinforcing means of teaching both the function and the form of these tacts, conversations can be successfully initiated by children with ASD.

Prolactin and cortisol regulate branchial claudin expression in Japanese medaka.

Several gill claudin (Cldn) tight junction proteins in Japanese medaka are regulated by salinity (cldn10 paralogs and cldn28b), while others are constitutively expressed (cldn27a, cldn28a and cldn30c). The role of the endocrine system in this regulation has yet to be understood. The in vitro effects of cortisol and prolactin on cldn expression in gill explant cultures were investigated in medaka. ncc2b and cftr were used as markers of specific ionocytes associated with freshwater- and seawater-acclimation, respectively. Concentration-response experiments were performed by overnight incubation with 0, 0.1, 1 and 10µgmL-1 cortisol or 0, 0.01, 0.1 and 1µgmL-1 ovine prolactin. Cortisol significantly up-regulated cftr, ncc2b, cldn10 paralogs, cldn27a and cldn30c from 1.2- to 5-fold control levels at 10µgmL-1. Cortisol had no effect on cldn28a and cldn28b. Prolactin had a concentration-dependent effect, decreasing expression of cftr (1µgmL-1, 2.2-fold) while increasing ncc2b (from 0.1µgmL-1, 6-7-fold). Prolactin up-regulated expression of 3 cldns: cldn28b (0.1 and 1µgmL-1), cldn10c and cldn10f (1µgmL-1), with up to 2-, 2.5- and 2-fold of control level, respectively. A combination experiment with both hormones showed that they act in synergy on cldn28b and have an additive effect on cftr, ncc2b, cldn10c and cldn10f. Our results showed that cortisol and prolactin are essential to maintain the expression of specific branchial claudins. This work also provides evidence that both hormones act directly on gill of medaka to modulate determinants of paracellular ion movement.

Windblown Pliocene diatoms and East Antarctic Ice Sheet retreat.

Marine diatoms in tillites along the Transantarctic Mountains (TAMs) have been used to suggest a diminished East Antarctic Ice Sheet (EAIS) during Pliocene warm periods. Updated ice-sheet modelling shows significant Pliocene EAIS retreat, creating marine embayments into the Wilkes and Aurora basins that were conducive to high diatom productivity and rapid accumulation of diatomaceous sediments. Here we show that subsequent isostatic uplift exposed accumulated unconsolidated marine deposits to wind erosion. We report new atmospheric modelling utilizing Pliocene climate and derived Antarctic landscapes indicating that prevailing mid-altitude winds transported diatoms towards the TAMs, dominantly from extensive emerged coastal deposits of the Aurora Basin. This result unifies leading ideas from competing sides of a contentious debate about the origin of the diatoms in the TAMs and their link to EAIS history, supporting the view that parts of the EAIS are vulnerable to relatively modest warming, with possible implications for future sea-level rise.

Effect of depression and anxiety on the success of pelvic floor muscle training for pelvic floor dysfunction.

The objective of this study was to determine the impact of the psychiatric symptoms of anxiety and depression, as assessed by validated questionnaires on the success of pelvic floor muscle training (PFMT). A prospective observational study was carried out by the Uro-gynaecological Physiotherapy Department at the Singleton Hospital, Swansea. A total of 108 consecutive women with pelvic floor dysfunction were referred for physiotherapy and admitted to the 6-month physiotherapy programme. They underwent subjective and objective assessments of their pelvic floor and psychological health at the beginning and end of the programme. A strong correlation was noted between the severity of anxiety and depression symptoms and the severity of their pelvic floor dysfunction. Following physiotherapy, apart from sexual function, all domains of pelvic floor dysfunction showed significant improvement. Based on the severity of their anxiety/depression symptoms, the patients were stratified into three groups. The group of patients that benefitted most had either no or only mild anxiety/depression. This study raises the question of whether a targeted approach should be undertaken for managing patients who, in addition to their pelvic floor dysfunction, demonstrate psychiatric symptoms.

The incidence, diagnostic clinical manifestations and severity of juvenile systemic lupus erythematosus in New Zealand Maori and Pacific Island children: the Starship experience (2000-2010).

OBJECTIVES: To describe the incidence, diagnostic clinical manifestations and severity of juvenile systemic lupus erythematosus (jSLE) in a cohort of New Zealand Maori and Pacific Island children compared to European children. METHODS: A chart review was conducted of children with jSLE seen by the Starship paediatric rheumatology and/or renal services between January 2000 and November 2010. Diagnostic clinical data and lupus nephritis data at anytime were collated while classic British Isles Lupus Assessment Group (BILAG) and Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores were derived retrospectively. RESULTS: Thirty-two children were diagnosed with jSLE with an annual incidence of 0.52 per 100,000 per year. Compared with European children (0.31 per 100,000 per year) the incidence of jSLE was higher among Maori and Pacific (0.67 per 100,000 per year, p=0.06) and significantly higher among Asian children (1.17 per 100,000 per year, p=0.01). Compared with European children, Maori and Pacific children were more frequently diagnosed with lupus nephritis (80% vs 40%, p=0.09) and severe (WHO class 4 or 5) renal lesions (60% vs 40%, p=0.43) at presentation. Similarly, at any time during the study, lupus nephritis (100% vs 40%, p=0.001) and severe (WHO class 4 or 5) renal lesions (73.3% vs 40%, p=0.12) were more frequent among Maori and Pacific compared with European children. Furthermore, retrospective BILAG assessment of diagnostic disease severity demonstrated that Maori and Pacific children experienced the majority of severe "Category A" disease (56.8% vs 22.7%, p=0.17) which was predominantly renal (73.3% vs 40%, p=0.12) in nature. CONCLUSIONS: This is the first description of the incidence and clinical manifestations of jSLE in a cohort of New Zealand children. Although limited by the small numbers involved it confirmed anecdotal suspicions that the incidence of jSLE among Maori, Pacific and Asian children is higher than European children. Lupus nephritis is also more frequent and severe in Maori and Pacific children.

Consideration of the degree of increase in urine metadrenalines provides superior specificity in the diagnosis of phaeochromocytoma than additional urine catecholamine measurement.

BACKGROUND: Measurement of fractionated plasma or urine metadrenalines is the recommended screening test in the diagnosis of phaeochromocytoma, with clinical cut-offs geared towards diagnostic sensitivity. Current practice at Salford Royal Hospital is to add urine catecholamines onto samples with raised urine metadrenalines, with the aim of adding specificity to a diagnosis of phaeochromocytoma. METHODS: This practice was reviewed by identifying a series of patients with raised urine metadrenalines who had catecholamines reflectively added. A total of 358 samples were identified from 242 patients, of which 228 had urine catecholamines measured. RESULTS: A diagnosis of 'phaeochromocytoma' (n = 41) or 'no phaeochromocytoma' (n = 90) was obtained in 131 of 228 patients, giving raised urine metadrenalines a positive predictive value for phaeochromocytoma of 31%. The finding of increased urine catecholamines in samples with raised urine metadrenalines increased specificity for phaeochromocytoma to 70%. However, 95% diagnostic specificity for phaeochromocytoma could be achieved by the introduction of a second cut-off for urine metadrenalines geared towards maximizing specificity. CONCLUSIONS: Consideration of the degree of increase in urine metadrenalines is a superior method of determining the likelihood of phaeochromocytoma than measurement of urine catecholamines.

Tracking of body mass indices over 2 years in Maori and European children.

BACKGROUND/OBJECTIVES: In 2002, the prevalence of overweight and obesity, defined by body mass index (BMI), was higher in New Zealand Maori (40%) and Pacific (60%) than in European (24%) children; however, this does not take into account interethnic differences in body composition. This study compared trajectories of anthropometric indices from 2004 to 2006 among 5- and 10-year-old Maori and European children. SUBJECTS/METHODS: In 2004 and then in 2006, 1244 children (639 boys and 605 girls) aged 5 and 10 years had height, weight and fat-free mass (FFM) by bioimpedance measured to derive measures of fat mass (FM), percentage body fat (%BF), FM index (FMI, FM/height(2)), FFM index (FFMI, FFM/height(2)), and s.d. scores for BMI and %BF and BMI categories by International Obesity Task Force criteria. RESULTS: Body composition and growth in Maori children is different from European children. Over 2 years, the BMI and %BF s.d. scores in both 5- and 10-year-old cohorts increased more in Maori children than in European children. The prevalence of overweight and obesity also increased within ethnicity and age group. The relative change of FMI and FFMI differed by age group, gender and ethnicity. In Maori girls, the magnitude of the changes between 10 and 12 years of age was most marked-the major contribution was from an increase in FMI. At 12 years, the mean %BF of Maori girls was 31.7% (95% confidence interval (CI): 30.1 and 33.3) compared with that of European girls (28.0%; 95% CI: 27.0 and 29.0). CONCLUSIONS: FM and FFM measures provide a more appropriate understanding of growth and body composition change in children than BMI, and vary with gender and ethnicity. Thus, FMI and FFMI should be tracked and compared among populations.

Comparison of conditioning impairments in children with Down syndrome, autistic spectrum disorders and mental age-matched controls.

BACKGROUND: This study investigated the relative ease of learning across four tasks suggested by an adaptation of Thomas's hierarchy of learning in children with Down syndrome, autism spectrum disorders and mental age-matched controls. METHODS: Learning trials were carried out to investigate observational learning, instrumental learning, reversal learning and conditional discrimination. RESULTS: The sample with autism spectrum disorders performed worse than the other two groups on the observational learning and conditional discrimination tasks, while the Down syndrome sample performed worse on the instrumental learning task. CONCLUSIONS: These findings are discussed in terms of there implications for reward-based educational intervention programmes.

Randomised controlled trial comparing oral and intravenous paracetamol (acetaminophen) plasma levels when given as preoperative analgesia.

Gastric absorption of oral paracetamol (acetaminophen) may be unreliable perioperatively in the starved and stressed patient. We compared plasma concentrations of parenteral paracetamol given preoperatively and oral paracetamol when given as premedication. Patients scheduled for elective ear; nose and throat surgery or orthopaedic surgery were randomised to receive either oral or intravenous paracetamol as preoperative medication. The oral dose was given 30 minutes before induction of anaesthesia and the intravenous dose given pre-induction. All patients were given a standardised anaesthetic by the same specialist anaesthetist who took blood for paracetamol concentrations 30 minutes after the first dose and then at 30 minute intervals for 240 minutes. Therapeutic concentrations of paracetamol were reached in 96% of patients who had received the drug parenterally, and 67% of patients who had received it orally. Maximum median plasma concentrations were 19 mg.l(-1) (interquartile range 15 to 23 mg.l(-1)) and 13 mg.l(-1) (interquartile range 0 to 18 mg.l(-1)) for the intravenous and oral group respectively. The difference between intravenous and oral groups was less marked after 150 minutes but the intravenous preparation gave higher plasma concentrations throughout the study period. It can be concluded that paracetamol gives more reliable therapeutic plasma concentrations when given intravenously.