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Individuals with obesity are vulnerable to low rates of preventive health screening. Veterans with obesity seeking bariatric surgery are also hypothesized to have gaps in preventive health screening. Evaluation in a multidisciplinary bariatric surgery clinic is a point of interaction with the healthcare system that could facilitate improvements in screening.This is a retrospective cohort study of 381 consecutive patients undergoing bariatric surgery at a Veterans Affairs Hospital from January 2010 to October 2021. Age- and sex-appropriate health screening rates were determined at initial referral to a multidisciplinary bariatric surgery clinic and at the time of surgery. Rates of guideline concordance at both time points were compared using McNemar's test. Univariate and multivariate analyses were performed to identify the risk factors for nonconcordance.Concordance with all recommended screening was low at initial referral and significantly improved by time of surgery (39.1%‒63.8%; p0.001). Screening rates significantly improved for HIV (p0.001), cervical cancer (p=0.03), and colon cancer (p0.001). Increases in BMI (p=0.005) and the number of indicated screening tests (p=0.029) were associated with reduced odds of concordance at initial referral. Smoking history (p=0.012) and increasing distance to the nearest Veterans Affairs Medical Center (p=0.039) were associated with reduced odds of change from nonconcordance at initial referral to concordance at the time of surgery.Rates of preventive health screening in Veterans with obesity are low. A multidisciplinary bariatric surgery clinic is an opportunity to improve preventive health screening in Veterans referred for bariatric surgery.

Dorsal and ventral medial entorhinal cortex (mEC) regions have distinct neural network firing patterns to differentially support functions such as spatial memory. Accordingly, mEC layer II dorsal stellate neurons are less excitable than ventral neurons. This is partly because the densities of inhibitory conductances are higher in dorsal than ventral neurons. Here, we report that T-type Ca2+ currents increase 3-fold along the dorsal-ventral axis in mEC layer II stellate neurons, with twice as much CaV3.2 mRNA in ventral mEC compared with dorsal mEC. Long depolarizing stimuli trigger T-type Ca2+ currents, which interact with persistent Na+ currents to elevate the membrane voltage and spike firing in ventral, not dorsal, neurons. T-type Ca2+ currents themselves prolong excitatory postsynaptic potentials (EPSPs) to enhance their summation and spike coupling in ventral neurons only. These findings indicate that T-type Ca2+ currents critically influence the dorsal-ventral mEC stellate neuron excitability gradient and, thereby, mEC dorsal-ventral circuit activity.

Lipidomics studies suffer from analytical and annotation challenges because of the great structural similarity of many of the lipid species. To improve lipid characterization and annotation capabilities beyond those afforded by traditional mass spectrometry (MS)-based methods, multidimensional separation methods such as those integrating liquid chromatography, ion mobility spectrometry, collision-induced dissociation and MS (LC-IMS-CID-MS) may be used. Although LC-IMS-CID-MS and other multidimensional methods offer valuable hydrophobicity, structural and mass information, the files are also complex and difficult to assess. Thus, the development of software tools to rapidly process and facilitate confident lipid annotations is essential. In this Protocol Extension, we use the freely available, vendor-neutral and open-source software Skyline to process and annotate multidimensional lipidomic data. Although Skyline ( https://skyline.ms/skyline.url ) was established for targeted processing of LC-MS-based proteomics data, it has since been extended such that it can be used to analyze small-molecule data as well as data containing the IMS dimension. This protocol uses Skyline's recently expanded capabilities, including small-molecule spectral libraries, indexed retention time and ion mobility filtering, and provides a step-by-step description for importing data, predicting retention times, validating lipid annotations, exporting results and editing our manually validated 500+ lipid library. Although the time required to complete the steps outlined here varies on the basis of multiple factors such as dataset size and familiarity with Skyline, this protocol takes ~5.5 h to complete when annotations are rigorously verified for maximum confidence.

The tourism industry makes a significant contribution to the socio-economic development of many countries of the world, due to its role in expanding the economic base, increasing foreign exchange earnings, developing small and medium-sized businesses, providing employment opportunities and improving the living standards of the population. The development of the tourism industry can be facilitated by its management on the basis of public-private partnerships. This form of cooperation helps to accelerate the solution of the country's strategic tasks, bringing significant social benefits to local communities. The purpose of the research was to study various forms of interaction between the state and business to solve the problems of developing the tourism industry in order to identify the optimal mechanism for public-private partnership for the development of the tourism industry in Kazakhstan. The paper studied the theoretical and methodological aspects and foreign experience in the formation and implementation of the mechanism of public-private partnership in the field of tourism. The article provides a comprehensive analysis of foreign experience in the creation, promotion and management of projects in the tourism industry on the principles of public-private partnership. The practical value is that the results of the study can be used in the development of state and regional programs for the development of the tourism industry in Kazakhstan.

Purpose: Our proof-of-concept study tested the feasibility of virtual testing using child assessments that were originally validated for in-person testing only. Method: Ten adult–child dyads were assigned to complete both in-person and virtual tests of language, cognition, and narratives. Child participants fell between the ages of 4 and 8 years; adult participants were speech-language clinicians or researchers with experience in administering child assessments. Half of child participants were Spanish–English bilinguals, and half were monolingual English speakers. Results: Results showed similar performance across in-person and virtual modalities on all assessments. Recommendations for adapting, administering, and scoring virtual measures with linguistically diverse children are discussed. Conclusions: Although additional research on virtual assessment is needed, our results open opportunities for appropriate remote assessment, particularly for bilingual children, who may not have in-person access to speech-language pathology services.

Voltage-gated calcium (CaV) channels form three subfamilies (CaV1–3). The CaV1 and CaV2 channels are heteromeric, consisting of an α1 pore-forming subunit, associated with auxiliary CaVβ and α2δ subunits. The α2δ subunits are encoded in mammals by four genes, CACNA2D1–4. They play important roles in trafficking and function of the CaV channel complexes. Here we report biallelic variants in CACNA2D1, encoding the α2δ-1 protein, in two unrelated individuals showing a developmental and epileptic encephalopathy. Patient 1 has a homozygous frameshift variant c.818_821dup/p.(Ser275Asnfs*13) resulting in nonsense-mediated mRNA decay of the CACNA2D1 transcripts, and absence of α2δ-1 protein detected in patient-derived fibroblasts. Patient 2 is compound heterozygous for an early frameshift variant c.13_23dup/p.(Leu9Alafs*5), highly probably representing a null allele and a missense variant c.626G>A/p.(Gly209Asp). Our functional studies show that this amino-acid change severely impairs the function of α2δ-1 as a calcium channel subunit, with strongly reduced trafficking of α2δ-1G209D to the cell surface and a complete inability of α2δ-1G209D to increase the trafficking and function of CaV2 channels. Thus, biallelic loss-of-function variants in CACNA2D1 underlie the severe neurodevelopmental disorder in these two patients. Our results demonstrate the critical importance and non-interchangeability of α2δ-1 and other α2δ proteins for normal human neuronal development.

A major barrier for the full utilization of metal additive manufacturing (AM) technologies is quality control. Additionally, in situ real time nondestructive monitoring is desirable due to the typical high value and low volume of components manufactured with metal AM. Depending on the application, characteristics such as the geometrical accuracy, porosity, defect size and content, and material properties are quantities of interest for in situ nondestructive evaluation (NDE). In particular, functionally tailored components made with hybrid processing require quantitative NDE of their microstructure and elastic properties. Ultrasonic NDE is able to quantify these relevant characteristics. In this work, an ultrasonic measurement system is used to collect in situ real time measurements during the manufacturing of samples made with a hybrid process, which combines directed energy deposition with milling. In addition to quantifying ultrasonic properties, the measurements are used to gather insight on other geometry, material, and process effects. The results show the utility of ultrasound to evaluate relevant properties during manufacturing of a functionalized material domain, while providing perspective on additional material evolution information obtained from ultrasonic signals.

Background Primary sclerosing cholangitis (PSC) is a well-described risk factor for the development of cholangiocarcinoma (CCA). Early detection of CCA in these patients is of great importance because it expands options for therapeutic interventions, including liver transplantation. Current diagnostic tests for the evaluation of biliary strictures are limited to biliary brushing (BB) cytology and fluorescence in situ hybridization (FISH). Next-generation sequencing (NGS) has become an important diagnostic tool in oncology and may be a useful tool for diagnosing CCA on BBs. It is not clear how NGS performs when it is added to BB cytology and FISH in patients with PSC. Methods This study reports the authors' experience with NGS performed as a prospective cotest with cytology and FISH on BBs obtained from 60 patients with PSC followed at Massachusetts General Hospital. A duct with malignancy was defined as a high-risk (HR) stricture with either high-grade dysplasia or CCA. Results NGS was better than FISH and cytology in detecting HR strictures, which showed multiple genetic mutations in all cases. NGS provided specific mutational information, and NGS results were reproducible in longitudinal samples. Conclusions Adding NGS to BB cytology and FISH in the evaluation of biliary strictures for patients with PSC may provide additional information that could help to inform clinical management.

Obesity is a critical problem in the equine industry, with ponies being at particularly high risk of developing obesity-related conditions such as metabolic syndrome and laminitis. The purpose of this study was to estimate the level of adiposity in an elite level of competitive show ponies, and to determine if adiposity was related to performance based on their model score (judged on conformation and appearance). Two researchers visually evaluated the body condition score (BCS) and cresty neck score (CNS) of 377 ponies at a national hunter competition. Thirty five percent of the ponies in the competition were considered obese, with BCS scores of 7 or higher. Mean BCS of the ponies was 6.7 ± 0.6 with a range of 5.25 - 8.25 and mean CNS was 2.8 ± 0.6, with a range of 1.75 - 4.5. Medium ponies had higher BCS (P0.0001) and higher CNS (P=0.015) than the large ponies. There was a tendency for a relationship between body condition score and the model score in all ponies (r = 0.08; P = 0.059), and this was stronger within the large ponies (r =0.20; P0.01). These findings show that elite competition ponies are dangerously overweight and that adiposity may influence performance in a judged event. This is of grave concern to the horse industry and needs to be addressed.

Male C1 canoe slalom athletes traditionally used cross transitions to move their paddle to the other side of the boat and off-side strokes to paddle on their non-dominant side. Conversely, female athletes often use a switching transition and on-side strokes on their non-dominant side. The purpose of this study was to use a computer model to assess the relation between cross- or switching techniques, and the relative strength (symmetry) of non-dominant compared to dominant side strokes to race times in C1 canoe slalom. We created a forward dynamics model to predict race times using stroke forces (from an indoor ergometer), drag forces (measured on-water), and probability distributions for stroke and transition times (measured from international canoe slalom competitions). The main effects from an ANOVA (

The ER Ca2+ channel ryanodine receptor 2 (RyR2) is required for maintenance of insulin content and glucose-stimulated insulin secretion, in part, via regulation of the protein IRBIT in the insulinoma cell line INS-1. Here, we examined store-operated and depolarization-dependent Ca2+entry using INS-1 cells in which either RyR2 or IRBIT were deleted. Store-operated Ca2+ entry (SOCE) stimulated with thapsigargin was reduced in RyR2KO cells compared to controls, but was unchanged in IRBITKO cells. STIM1 protein levels were not different between the three cell lines. Basal and stimulated (500 μM carbachol) phospholipase C (PLC) activity was also reduced specifically in RyR2KO cells. Insulin secretion stimulated by tolbutamide was reduced in RyR2KO and IRBITKO cells compared to controls, but was potentiated by an EPAC-selective cAMP analog in all three cell lines. Cellular PIP2 levels were increased and cortical f-actin levels were reduced in RyR2KO cells compared to controls. Whole-cell Cav channel current density was increased in RyR2KO cells compared to controls, and barium current was reduced by acute activation of the lipid phosphatase pseudojanin preferentially in RyR2KO cells over control INS-1 cells. Action potentials stimulated by 18 mM glucose were more frequent in RyR2KO cells compared to controls, and insensitive to the SK channel inhibitor apamin. Taken together, these results suggest that RyR2 plays a critical role in regulating PLC activity and PIP2 levels via regulation of SOCE. RyR2 also regulates β-cell electrical activity by controlling Cav current density and SK channel activation.

Vendor-independent software tools for quantification of small molecules and metabolites are lacking, especially for targeted analysis workflows. Skyline is a freely available, open-source software tool for targeted quantitative mass spectrometry method development and data processing with a ten-year history supporting 6 major instrument vendors. Designed initially for proteomic analysis, we describe the expansion of Skyline to data for small molecule analysis, including selected reaction monitoring (SRM), high-resolution mass spectrometry (HRMS), and calibrated quantification. This fundamental expansion of Skyline from a peptide-sequence centric tool to a molecule-centric tool makes it agnostic to the source of the molecule while retaining Skyline features critical for workflows in both peptide and more general biomolecular research. The data visualization and interrogation features already available in Skyline - such as peak picking, chromatographic alignment, and transition selection - have been adapted to support small molecule data, including metabolomics. Herein, we explain the conceptual workflow for small molecule analysis using Skyline, demonstrate Skyline performance benchmarked against a comparable instrument vendor software tool, and present additional real-world applications. Further, we include step-by-step instructions on using Skyline for small molecule quantitative method development and data analysis on data acquired with a variety of mass spectrometers from multiple instrument vendors.

The implication of lipid dysregulation in diseases, toxic exposure outcomes, and inflammation has brought great interest to lipidomic studies. However, lipids have proven to be analytically challenging due to their highly isomeric nature and vast concentration ranges in biological matrices. Therefore, multidimensional techniques such as those integrating liquid chromatography, ion mobility spectrometry, collision-induced dissociation, and mass spectrometry (LC-IMS-CID-MS) have been implemented to separate lipid isomers as well as provide structural information and increased identification confidence. These data sets are however extremely large and complex, resulting in challenges for data processing and annotation. Here, we have overcome these challenges by developing sample-specific multidimensional lipid libraries using the freely available software Skyline. Specifically, the human plasma library developed for this work contains over 500 unique lipids and is combined with adapted Skyline functions such as indexed retention time (iRT) for retention time prediction and IMS drift time filtering for enhanced selectivity. For comparison with other studies, this database was used to annotate LC-IMS-CID-MS data from a NIST SRM 1950 extract. The same workflow was then utilized to assess plasma and bronchoalveolar lavage fluid (BALF) samples from patients with varying degrees of smoke inhalation injury to identify lipid-based patient prognostic and diagnostic markers.

Purpose Producing a steady cadence and power while cycling results in fairly consistent average pedal forces for every revolution, though small fluctuations about an average force do occur. This force can be generated by several combinations of muscles, each with slight fluctuations in excitation for every pedal cycle. Fluctuations such as these are commonly thought of as random variation about average values. However, research into fluctuations of stride length and stride time during walking shows information can be contained in the order of fluctuations. This order, or structure, is thought to reveal underlying motor control strategies. Previously, we found persistent structure in the fluctuations of EMG signals during cycling using Entropic Half-Life (EnHL) analysis. These EMG signals contained fluctuations across multiple timescales, such as those within a burst of excitation, between the burst and quiescent period of a cycle, and across multiple cycles. It was not clear which sources of variation contributed to the persistent structure in the EMG. Methods In this study, we manipulated variation at different timescales in EMG intensity signals to identify the sources of structure observed during cycling. Nine participants cycled at a constant power and cadence for 30 minutes while EMG was collected from six muscles of the leg. Results We found persistent structure across multiple pedal cycles of average EMG intensities, as well as average pedal forces and durations. Additionally, we found the EnHL did not quantify fluctuations within a burst of EMG intensity; instead, it detected unstructured variation between the burst and quiescent period within a cycle. Conclusions The persistent structure in average EMG intensities suggests that fluctuations in muscle excitation are regulated from cycle to cycle.

Summary Skyline is a Windows application for targeted mass spectrometry method creation and quantitative data analysis. Like most graphical user interface (GUI) tools, it has a complex user interface with many ways for users to edit their files which makes the task of logging user actions challenging and is the reason why audit logging of every change is not common in GUI tools. We present an object comparison-based approach to audit logging for Skyline that is extensible to other GUI tools. The new audit logging system keeps track of all document modifications made through the GUI or the command line and displays them in an interactive grid. The audit log can also be uploaded and viewed in Panorama, a web repository for Skyline documents that can be configured to only accept documents with a valid audit log, based on embedded hashes to protect log integrity. This makes workflows involving Skyline and Panorama more reproducible. Availability and implementation Skyline is freely available at https://skyline.ms. Supplementary information Supplementary data are available at Bioinformatics online.

Porous graphitized carbon (PGC) based chromatography achieves high-resolution separation of glycan structures released from glycoproteins. This approach is especially valuable when resolving structurally similar isomers and for discovery of novel and/or sample-specific glycan structures. However, the implementation of PGC-based separations in glycomics studies has been limited because system-independent retention values have not been established to normalize technical variation. To address this limitation, this study combined the use of hydrolyzed dextran as an internal standard and Skyline software for post-acquisition normalization to reduce retention time and peak area technical variation in PGC-based glycan analyses. This approach allowed assignment of system-independent retention values that are applicable to typical PGC-based glycan separations and supported the construction of a library containing >300 PGC-separated glycan structures with normalized glucose unit (GU) retention values. To enable the automation of this normalization method, a spectral MS/MS library was developed of the dextran ladder, achieving confident discrimination against isomeric glycans. The utility of this approach is demonstrated in two ways. First, to inform the search space for bioinformatically predicted but unobserved glycan structures, predictive models for two structural modifications, core-fucosylation and bisecting GlcNAc, were developed based on the GU library. Second, the applicability of this method for the analysis of complex biological samples is evidenced by the ability to discriminate between cell culture and tissue sample types by the normalized intensity of N-glycan structures alone. Overall, the methods and data described here are expected to support the future development of more automated approaches to glycan identification and quantitation.

International audience

SummaryThe medial entorhinal cortex (mEC) plays a salient role in physiological processes such as spatial cognition and spatial coding. mEC layer II stellate neurons, in particular, influence these processes. Interestingly, ventral and dorsal stellate neurons diversely affect these processes and have distinct intrinsic membrane properties and action potential firing patterns. Little, though, is known about how ventral stellate neuron intrinsic excitability is regulated. We show that ventral stellate neurons predominantly possess T-type Ca2+ currents encoded by CaV3.2 subunits, with dorsal stellate neurons having small or no currents. Further, twice as much CaV3.2 mRNA was present in ventral than dorsal mEC. In line with T-type, CaV3.2 Ca2+ current biophysical properties, depolarising stimuli activated these currents in ventral, but not dorsal, neurons. Here, these currents acted in concert with persistent Na+ currents to elevate input resistance and tonic action potential firing. CaV3.2 currents also enhanced excitatory post-synaptic potential decay and integration solely in ventral neurons. These results reveal that CaV3.2 currents, together with persistent Na+ currents, impart the characteristic intrinsic membrane and firing properties of ventral stellate neurons. This signifies that specific voltage-gated conductances distinctly affect ventral and dorsal mEC stellate neuron activity and functions such as spatial memory and spatial navigation.

IntroductionDirect-to-family clinical trials efficiently provide data while reducing the participation burden for children and their families. Although these trials can offer significant advantages over traditional clinical trials, the process of designing and implementing direct-to-family studies is poorly defined, especially in children with rheumatic disease. This paper provides lessons learnt from the design and implementation of a self-controlled, direct-to-family pilot trial aimed to evaluate the effects of a medication management device on adherence to hydroxychloroquine in paediatric SLE.MethodsSeveral design features accommodate a direct-to-family approach. Participants meeting eligibility criteria from across the USA were identified a priori through a disease registry, and all outcome data are collected remotely. The primary outcome (medication adherence) is evaluated using electronic medication event-monitoring, plasma drug levels, patient questionnaires and pill counts. Secondary and exploratory endpoints include (1) lupus disease activity measured by a remote SLE Disease Activity Index examination and the Systemic Lupus Activity Questionnaire; and (2) hydroxychloroquine pharmacokinetics and pharmacodynamics. Recruitment of the initial target of 20 participants was achieved within 10 days. Due to initial recruitment success, enrolment was increased to 26 participants. Additional participants who were interested were placed on a waiting list in case of dropouts during the study.Discussion and disseminationDirect-to-family trials offer several advantages but present unique challenges. Lessons learnt from the protocol development, design, and implementation of this trial will inform future direct-to-family trials for children and adults with rheumatic diseases. Additionally, the data collected remotely in this trial will provide critical information regarding the accuracy of teleresearch in lupus, the impact of adherence to hydroxychloroquine on disease activity and a pharmacokinetic analysis to inform paediatric-specific dosing of hydroxychloroquine.Trial registration numberClinicalTrials.gov Registry (NCT04358302).

Insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) are important biomarkers in research and diagnosis of growth disorders. Quantitative analysis is performed using various ligand-binding assays or enzymatic digestion LC-MS/MS methods, whose widespread adoption is hampered by time-consuming sample preparation procedures. We present a simple and fast antibody-free LC-MS/MS method for the quantification of intact IGF-1 and IGF-2 in human plasma. The method requires 50 μL of plasma and uses fully 15N-labelled IGF-1 as internal standard. It features trifluoroethanol (TFE)-based IGF/IGF-binding protein complex dissociation and a two-step selective protein precipitation workflow, using 5% acetic acid in 80/20 acetone/acetonitrile (precipitation 1) and ice-cold ethanol (precipitation 2). Detection of intact IGF-1 and IGF-2 is performed by means of a Waters XEVO TQ-S triple quadrupole mass spectrometer in positive electrospray ionisation (ESI+) mode. Lower limits of quantification were 5.9 ng/mL for IGF-1 and 8.4 ng/mL for IGF-2. Intra-assay imprecision was below 4.5% and inter-assay imprecision was below 5.8% for both analytes. An excellent correlation was found between nominal and measured concentrations of the WHO reference standard for IGF-1. Comparison with the IDS-iSYS IGF-1 immunoassay showed good correlation (R2 > 0.97), although a significant bias was observed with the immunoassay giving substantially higher concentrations. The LC-MS/MS method described here allows for reliable and simultaneous quantification of IGF-1 and IGF-2 in plasma, without the need for enzymatic digestion. The method can be readily implemented in clinical mass spectrometry laboratories and has the potential to be adapted for the analysis of different similarly sized peptide hormones. Graphical abstract

Purpose Maximally decreasing portal pressures with transjugular intrahepatic portosystemic shunt (TIPS) is associated with improved ascites control but also increased encephalopathy incidence. Since splenic venous flow contributes to portal hypertension, we assessed if combining small-diameter TIPS with splenic artery embolization could improve ascites while minimizing encephalopathy. Methods Fifty-five patients underwent TIPS creation for refractory ascites. Subjects underwent creation of 8 mm TIPS followed by proximal splenic artery embolization (group A, n=8), or of 8 mm (group B, n=6) or 10 mm TIPS (group C, n=41) without splenic embolization. Data were retrospectively reviewed. Results In group A, median portosystemic gradient decreased from 19 mmHg to 9 mmHg after TIPS, and 8 mmHg after subsequent splenic artery embolization. In groups B and C, gradient decreased from 15 mmHg to 8 mmHg and 16 mmHg to 6 mmHg. All patients except for one in group A and two in C had greater than 50% reduction in the number of paracenteses in 3 months. Any postprocedural encephalopathy incidence was 62%, 50%, 83% in groups A, B, and C, respectively. Overall, 20% of subjects with 10 mm TIPS required TIPS reduction/closure compared to 7% of subjects with 8 mm TIPS. Conclusion We found that 8 mm diameter TIPS provided similar ascites control compared to 10 mm TIPS regardless of splenic embolization. While more patients with 10 mm TIPS required reduction/closure for severe encephalopathy, the study was underpowered for definitive assessment. Splenic embolization might have the potential to further decrease portosystemic gradient and ascites as an alternative to dilation of TIPS to 10 mm minimizing the risk of encephalopathy, but larger studies are warranted.

Genetically-encoded fluorescent sensors have been widely used to illuminate secretory vesicle dynamics and the vesicular lumen, including Zn(2+) and pH, in living cells. However, vesicular sensors have a tendency to mislocalize and are susceptible to the acidic intraluminal pH. In this study, we performed a systematic comparison of five different vesicular proteins to target the fluorescent protein mCherry and a Zn(2+) FRET sensor to secretory vesicles. We found that motifs derived from vesicular cargo proteins, including Chromogranin A (CgA), target vesicular puncta with greater efficacy than transmembrane proteins. To characterize vesicular Zn(2+) levels, we developed CgA-Zn(2+) FRET sensor fusions with existing sensors ZapCY1 and eCALWY-4, and characterized subcellular localization and the influence of pH on sensor performance. We simultaneously monitored Zn(2+) and pH in individual secretory vesicles by leveraging the acceptor fluorescent protein as a pH sensor and found that pH influenced FRET measurements in situ. While unable to characterize vesicular Zn(2+) levels at the single vesicle level, we were able to monitor Zn(2+) dynamics in populations of vesicles and detected high vesicular Zn(2+) in MIN6 cells compared to lower levels in the prostate cancer cell line LnCaP. The combination of CgA-ZapCY1 and CgA-eCALWY-4 allows for measurement of Zn(2+) from pM to nM ranges.

UNSTRUCTURED Excess weight is a major risk factor for chronic diseases. In Australia, over 60% of adults are overweight or obese. The overconsumption of energy-dense nutrient-poor (EDNP) foods and low physical activity (PA) levels are key factors contributing to population obesity. New cost-effective approaches to improve population diet and PA behaviors are needed. This 1-year randomized controlled trial (6-month intervention and 6-month follow-up) aims to investigate whether a tailored intervention using mobile technology can improve diet and PA behaviors leading to weight loss in adults (aged 18-65 years) who are overweight or obese and recruited through a social marketing campaign (LiveLighter). All eligible participants will provide data on demographics and lifestyle behaviors online at baseline, 6 months, and 12 months. Using two-stage randomization, participants will be allocated into one of three conditions (n=200 per group): tailored feedback delivered via email at seven time points, informed by objective dietary (mobile food record app) and activity (wearable activity monitor) assessment; active control receiving no tailored feedback, but undergoing the same objective assessments as tailored feedback; and online control receiving no tailored feedback or objective assessments. Primary outcome measures at 6 and 12 months are changes in body mass, EDNP food and beverage consumption, and daily moderate-to-vigorous PA (measured via accelerometry). Secondary outcomes include change in fruit and vegetable consumption, daily sedentary behaviors, and cost effectiveness. Enrolment commenced in August 2017. Primary outcomes at 12 months will be available for analysis from September 2019. Tailored email feedback provided to individuals may deliver a cost-effective strategy to overcome existing barriers to improving diet and PA. If found to be successful and cost effective, upscaling this intervention for inclusion in larger-scale interventions is highly feasible. Australian New Zealand Clinical Trials Registry ACTRN12617000554369; https://www.anzctr.org.au /Trial/Registration/TrialReview.aspx?id=371325&isReview=true.

Background and aims Coronavirus disease 2019 (COVID-19), the illness caused by the SARS-CoV-2 virus, is rapidly spreading throughout the world. Hospitals and healthcare providers are preparing for the anticipated surge in critically ill patients, but few are wholly equipped to manage this new disease. The goals of this document are to provide data on what is currently known about COVID-19, and how it may impact hepatologists and liver transplant providers and their patients. Our aim is to provide a template for the development of clinical recommendations and policies to mitigate the impact of the COVID-19 pandemic on liver patients and healthcare providers. Approach and results This article discusses what is known about COVID-19 with a focus on its impact on hepatologists, liver transplant providers, patients with liver disease, and liver transplant recipients. We provide clinicians with guidance for how to minimize the impact of the COVID-19 pandemic on their patients' care. Conclusions The situation is evolving rapidly, and these recommendations will need to evolve as well. As we learn more about how the COVID-19 pandemic impacts the care of patients with liver disease, we will update the online document available at https://www.aasld.org/about-aasld/covid-19-and-liver.

Background Coronavirus disease 2019 (COVID-19) is associated with elevated liver biochemistries in approximately half of hospitalized patients, with many possible etiologies. Aim To assess agreement on the etiology of abnormal liver biochemistries and diagnostic recommendations in COVID-19. Methods Twenty hepatology consultations were reviewed by three senior hepatologists who provided a differential diagnosis and diagnostic recommendations. Kappa agreement on the primary etiology was calculated. Results Kappa agreement between hepatologists on the primary etiology of elevated liver biochemistries was 0.10 (p = 0.03). Agreement was greater around drug-induced liver injury 0.51 (p

Objective:To examine demographic and behavioural correlates of frequent consumption of fast food among Australian secondary school students and explore the associations between fast food consumption and social/environmental factors.Design:Cross-sectional survey using a web-based self-report questionnaire.Setting:Secondary schools across all Australian states and territories.Participants:Students aged 12–17 years participating in the 2012–2013 National Secondary Students’ Diet and Activity survey (n 8392).Results:Overall, 38 % of students surveyed reported consuming fast food at least weekly. Being male, residing in lower socio-economic areas and metropolitan locations, having more weekly spending money and working at a fast food outlet were all independently associated with consuming fast food once a week or more, as were several unhealthy eating (low vegetable intake and high sugary drink and snack food intake) and leisure (low physical activity and higher commercial television viewing) behaviours and short sleep duration. Frequent fast food consumption and measured weight status were unrelated. Students who agreed they go to fast food outlets with their family and friends were more likely to report consuming fast food at least weekly, as were those who usually ‘upsize’ their fast food meals and believe fast food is good value for money.Conclusions:These results suggest that frequent fast food consumption clusters with other unhealthy behaviours. Policy and educational interventions that reach identified at-risk groups are needed to reduce adolescent fast food consumption at the population level. Policies placing restrictions on the portion sizes of fast food may also help adolescents limit their intake.