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Several indicators reflect the quality of care within emergency departments (ED). The length of stay (LOS) of emergency patients represents one of the most important performance measures. Determinants of LOS have not yet been evaluated in large cohorts in Germany. This study analyzed the fixed and influenceable determinants of LOS by evaluating data from the German Emergency Department Data Registry (AKTIN registry). We performed a retrospective evaluation of all adult (age ≥ 18 years) ED patients enrolled in the AKTIN registry for the year 2019. Primary outcome was LOS for the whole cohort; secondary outcomes included LOS stratified by (1) patient-related, (2) organizational-related and (3) structure-related factors. Overall, 304,606 patients from 12 EDs were included. Average LOS for all patients was 3 h 28 min (95% CI 3 h 27 min–3 h 29 min). Regardless of other variables, patients admitted to hospital stayed 64 min longer than non-admitted patients. LOS increased with patients’ age, was shorter for walk-in patients compared to medical referral, and longer for non-trauma presenting complaints. Relevant differences were also found for acuity level, day of the week, and emergency care levels. We identified different factors influencing the duration of LOS in the ED. Total LOS was dependent on patient-related factors (age), disease-related factors (presentation complaint and triage level), and organizational factors (weekday and admitted/non-admitted status). These findings are important for the development of management strategies to optimize patient flow through the ED and thus to prevent overcrowding.

Zusammenfassung Hintergrund Die Notfallversorgung befindet sich im Umbruch. In Notaufnahmen werden Patienten ausgehend von Symptomen und Dringlichkeit versorgt; dies bildet sich jedoch in den etablierten Routinedaten der gesetzlichen Sozialversicherung nicht ab. Ziel des AKTIN-Projekts war der Aufbau einer datenschutzkonformen Registerinfrastruktur zur Nutzung von klinischen Routinedaten aus Notaufnahmen. Methoden Über eine standardisierte Schnittstelle werden aus den verschiedenen Dokumentationssystemen kontinuierlich Daten der Notaufnahmebehandlung in ein lokales Data Warehouse exportiert. Dort stehen sie sowohl für lokale Nutzungen, wie interne Berichte und Qualitätsmanagement, als auch gleichzeitig datenschutzkonform für multizentrische Auswertungen zur Verfügung. Anhand der Registerpopulation wird die Ersteinschätzung sowie die Erhebung von Vitalparametern in Abhängigkeit von Vorstellungsgründen für einen 12-Monats-Zeitraum analysiert. Ergebnisse Für den Zeitraum 04/2018 bis 03/2019 wurden 436.149 gültige Fälle aus 15 Notaufnahmen übermittelt. In 86,0 % der Fälle ist eine Ersteinschätzung dokumentiert. Diese fand in 70,5 % innerhalb von 10 min nach Ankunft des Patienten statt. In 10 Kliniken wird regelhaft (82,3 %) ein Vorstellungsgrund erfasst. Die Erfassung von Vitalparametern variiert plausibel zwischen den Vorstellungsgründen. Schlussfolgerung Das AKTIN-Notaufnahmeregister bietet einen zeitnahen Einblick in das Versorgungsgeschehen der Notaufnahmen ohne zusätzlichen Dokumentationsaufwand und unabhängig vom primären IT-System, Kostenträger, Fallart und Abrechnungsmodus. Die Vorgaben des Gemeinsamen Bundesausschusses zur Ersteinschätzung werden weitgehend umgesetzt. Durch die Etablierung von standardisierten Vorstellungsgründen werden symptombasierte Analysen und Gesundheitssurveillance ermöglicht.

The complexity of emergency cases and the number of emergency patients have increased dramatically. Due to a reduced or even missing specialist medical staff in the emergency departments (EDs), medical knowledge is often used without professional supervision for the diagnosis. The result is a failure in diagnosis and treatment, even death in the worst case. Secondary: high expenditure of time and high costs. Using accurate patient data from the German national registry of the medical emergency departments (AKTIN-registry, Home - Notaufnahmeregister (aktin.org)), the most 20 frequent diagnoses were selected for creating explainable artificial intelligence (XAI) models as part of the ENSURE project (ENSURE (umg.eu)). 137.152 samples and 51 features (vital signs and symptoms) were analyzed. The XAI models achieved a mean area under the curve (AUC) one-vs-rest of 0.98 for logistic regression (LR) and 0.99 for the random forest (RF), and predictive accuracies of 0.927 (LR) and 0.99 (RF). Based on its grade of explainability and performance, the best model will be incorporated into a portable CDSS to improve diagnoses and outcomes of ED treatment and reduce cost. The CDSS will be tested in a clinical pilot study at EDs of selected hospitals in Germany.

Purpose Early detection of bleeding is important for managing trauma cases in the emergency department (ED). Several trauma suites are equipped with computed tomography (CT) scanners to reduce the time to CT. In the last decade, sliding gantry CT has been implemented in trauma suites, highlighting conventional techniques' advantages. We investigated the change in the time to CT and the challenges faced during the implementation. Methods Trauma suite treatments with a conventional CT scanner between January and December 2016 formed the control group. From January to April 2017, trauma suites were modified, and treatment was outsourced to an interim trauma suite. By May 2017, trauma suites were equipped with a sliding gantry CT scanner. Treatments from May to July 2017 formed the transition group, and those from August to December 2017 formed the routine use group. We evaluated the time to CT in all groups and considered the reasons for the delays in the transition and routine use groups. Results On sliding gantry CT implementation, although time to CT remained unaffected in the transition group, it significantly reduced in the routine use group, independent of injury severity score. The incidence of cable management problems was significantly higher in the latter group. Conclusions We have demonstrated a decrease in the time to CT with the implementation of a sliding gantry CT. However, due to a higher number of cable management problems in the routine use group, we recommend regular refresher team training with routine use.

Neben einem Anstieg der Fallzahl in deutschen Notfallaufnahmen andert sich dort auch das Spektrum der vorstelligen Patienten. Statt einer Versorgung im vertragsarztlichen Sektor bevorzugen Patienten im akuten Krankheitsfall zunehmend die Notfallaufnahme als Anlaufstelle. Diese Arbeit untersucht das Inanspruchnahmeverhalten sowie dazugehorige Motive von Notfallaufnahmepatienten mit niedriger Dringlichkeit. Anonyme schriftliche Befragung in der Notfallaufnahme des Klinikums Wolfsburg zwischen 12/2015 und 03/2016 mit deskriptiver Auswertung. Teilnahmeberechtigt waren alle Patienten mit niedriger Dringlichkeit (Manchester-Triage-System (MTS), Stufen blau und grun). 81,5 % der Befragten (729 auswertbare von 7000 ausgegebenen Fragebogen) stellte sich zwischen 8:00 und 17:00 Uhr in der Notfallaufnahme vor, 70,1 % waren „Selbstvorsteller“. Am haufigsten (48,3 %) wurde eine vermutete bessere Versorgung als Motiv fur die Inanspruchnahme genannt. 67,8 % der Befragten schatzten sich entgegen der MTS-Stufe als mittelschweren bis lebensbedrohlichen Notfall ein. Als alternative Anlaufstelle wurden 49,2 % eine benachbarte Klinik in der Region wahlen. Die selbsteingeschatzte Behandlungsdringlichkeit der Patienten divergiert mit der Ersteinschatzung nach MTS. Bei akuter Angst um die Gesundheit werden vertragsarztliche Angebote, wie die Notfallpraxis der Kassenarztlichen Vereinigung in unmittelbarer Nahe, unzureichend genutzt. Bisherige Ansatze der Patientensteuerung scheinen in dem von uns untersuchten Setting wenig erfolgreich. Eine Starkung der Notfallaufnahmen als alleinige Anlaufstelle 24/7 bei gleichzeitiger Abschaffung paralleler Versorgungsstrukturen sollte diskutiert werden.

Talar neck and body fractures are rare. Major posttraumatic complications with a potential reduction in the quality of life are arthrosis and necrosis due to the specific vascular supply. The aim of the study was to evaluate mid-term results of surgery for talar fractures of neck and body. Parameters that potentially affected/influenced treatment outcomes were analysed exploratively.24 patients with 24 talar neck and body fractures (Marti type II n = 9, type III n = 12, type IV n = 3) were retrospectively examined for radiological and clinical functional outcomes. The independent parameters evaluated included age ( 40, ≥ 40 years), sex (male, female), general overall extent of injury (polytrauma/multiple injuries/multiple fractures of the extremities, additional injuries to the same foot, isolated talus fracture), soft tissue damage (open, closed), surgical latency ( 6, ≥ 6 h), fracture classification/displacement (undisplaced [= Marti II], displaced [= Marti III, IV]) and fracture type (talar body, neck fracture). The potential influencing parameters were analysed by univariate analyses.With an average follow-up of 8.7 years (1,25 - 16 years) the AOFAS score was 71.4 ± 22.9 points, the Foot Function Index score 35.9 ± 28.3 points; the physical and mental component summary scores of the Short Form 36, version 2, was 43.8 ± 10.9 and 47.4 ± 13.6 points (mean ± standard deviation), respectively. Thus, the patient reported physical health of the patients was slightly reduced compared to the German population, while the mental health remained largely unaffected. Two patients developed partial avascular necrosis (8%), 10 patients developed osteoarthritis (42%). Of the independent parameters, only the general overall extent of injury showed a significant influence on osteoarthritis (p = 0.002). In the evaluation of undisplaced (n = 9) and displaced (n = 15) fractures, surgical treatment after more than 6 hours did not result in a worse outcome.The clinical outcome of internal fixation of talar neck and body fractures can be classified as good. In the study group, there was no correlation between the occurrence of arthrosis and the Marti fracture classification.Zentrale Talusfrakturen (Hals- und Körperfrakturen) sind selten. Als wesentliche posttraumatische Komplikationen mit potenzieller Minderung der Lebensqualität gelten die Arthrose sowie die Nekrose aufgrund der besonderen Gefäßversorgung. Studienziel war die Evaluation mittelfristiger Ergebnisse nach osteosynthetischer Versorgung einschließlich explorativer Analyse potenzieller Einflussparameter auf die Behandlungsergebnisse.24 Patienten mit 24 operierten zentralen Talusfrakturen (Typ II nach Marti n = 9, Typ III n = 12, Typ IV n = 3) wurden retrospektiv radiologisch und klinisch nachuntersucht. Betrachtet wurden die Einflussgrößen Lebensalter ( 40, ≥ 40 Jahre), Geschlecht (männlich, weiblich), allgemeines Gesamtverletzungsausmaß (Polytrauma/Mehrfachverletzung/multiple Extremitätenfrakturen, weitere Verletzungen am gleichen Fuß, isolierte Talusfraktur), Weichteilschaden (offen, geschlossen), OP-Latenz ( 6, ≥ 6 h) sowie Frakturklassifikation/Dislokation (undisloziert [= Marti II], disloziert [= Marti III, IV]) und die Frakturlokalisation (Körper-, Halsfraktur). Die potenziellen Einflussgrößen wurden mittels univariater Analysen untersucht.Das mittlere Follow-up lag bei 8,7 Jahren (1,25 – 16 Jahre). Im Studienkollektiv betrug der AOFAS 71,4 ± 22,9 Punkte, der Fuß-Funktions-Index 35,9 ± 28,3 Punkte; die körperliche Summenskala und psychische Summenskala des Short Form-36 Version 2 lag bei 43,8 ± 10,9 und 47,4 ± 13,6 Punkten (Mittelwert ± Standardabweichung). Somit bewerteten die Patienten ihre körperliche Gesundheit etwas schlechter als der deutsche Durchschnitt, die psychische Komponente dagegen blieb weitestgehend unbeeinträchtigt. Zwei Patienten entwickelten eine partielle Nekrose (8%), 10 Patienten eine Arthrose (42%). Von den Einflussgrößen zeigte einzig das allgemeine Gesamtverletzungsausmaß einen signifikanten Einfluss auf die Arthrose (p = 0,002). In der Gesamtauswertung undislozierter (n = 9) und dislozierter (n = 15) Frakturen ergab die operative Versorgung nach über 6 h kein signifikant schlechteres Outcome.Die klinischen Ergebnisse nach osteosynthetischer Versorgung sind gut. In unserem Studienkollektiv ergab sich keine signifikante Korrelation zwischen dem Auftreten einer Arthrose und der Frakturklassifikation nach Marti.

Digitalization and standardization of documentation in medicine are increasingly progressing. In the decision of the Federal Joint Committee (G-BA) for a staged system of emergency structures in hospitals and in the report of the expert committee for development in the healthcare system on needs-oriented guidance of healthcare, the actuality of the topic is underlined with concrete naming of future tasks. The section on emergency admission protocols of the German Interdisciplinary Association of Intensive and Emergency Care Medicine (DIVI) has been working for years on this topic and has repeatedly reported on the progress of the topic in clinical emergency care. Standardization and digitalization represent the foundation for health services research spread across locations as well as the possibility for benchmarking. Digitalization makes the secondary use of primary clinical routine data possible. Digitalization decreases redundancies of data transmission by avoiding manual data input in, for example registers.

Introduction Quality of emergency department (ED) care affects patient outcomes substantially. Quality indicators (QIs) for ED care are a major challenge due to the heterogeneity of patient populations, health care structures and processes in Germany. Although a number of quality measures are already in use, there is a paucity of data on the importance of these QIs on medium-term and long-term outcomes. The evaluation of outcome relevance of quality indicators in the emergency department study (ENQuIRE) aims to identify and investigate the relevance of QIs in the ED on patient outcomes in a 12-month follow-up. Methods and analysis The study is a prospective non-interventional multicentre cohort study conducted in 15 EDs throughout Germany. Included are all patients in 2019, who were ≥18 years of age, insured at the Techniker Krankenkasse (statutory health insurance (SHI)) and gave their written informed consent to the study. The primary objective of the study is to assess the effect of selected quality measures on patient outcome. The data collected for this purpose comprise medical records from the ED treatment, discharge (claims) data from hospitalised patients, a patient questionnaire to be answered 6–8 weeks after emergency admission, and outcome measures in a 12-month follow-up obtained as claims data from the SHI. Descriptive and analytical statistics will be applied to provide summaries about the characteristics of QIs and associations between quality measures and patient outcomes. Ethics and dissemination Approval of the leading ethics committee at the Medical Faculty of the University of Magdeburg (reference number 163/18 from 19 November 2018) has been obtained and adapted by responsible local ethics committees. The findings of this work will be disseminated by publication of peer-reviewed manuscripts and presentations as conference contributions (abstracts, poster or oral presentations). Moreover, results will be discussed with clinical experts and medical associations before being proposed for implementation into the quality management of EDs. Trial registration number German Clinical Trials Registry (DRKS00015203); Pre-results.

Purpose To improve quality of trauma room management, intra- and inter-hospital benchmarking are important tools. However, primary data quality is crucial for benchmarking reliability. In this study, we analyzed the effect of a medical documentation assistant on documentation completeness in trauma room management in comparison to documentation by physicians involved in direct patient treatment. Methods We included all patients treated in the trauma room from 2016/01/01 to 2016/12/31 that were documented with the trauma module of the German Emergency Department Medical Record V2015.1. We divided the data into documentation by medical documentation assistant (DA, 07:00 to 17:00), physician in daytime (PD, 07:00 to 17:00), and physician at night (PN, 17:00 to 07:00). Data were analyzed for completeness (primary outcome parameter) as well as diagnostic intervals. Results There was a significant increase in complete recorded data for DA (74.5%; IQR 14.5%) compared to PD (26.9%; IQR 18.7%; p p p = 0.007) or PN (24.5 min; IQR 10.0; p = 0.001). Conclusion In presence of a qualified medical documentation assistant, data completeness and time to WBCT improved significantly. Therefore, utilizing a professional DA in the trauma room appears beneficial for data quality and time management.

The German Emergency Department Data Registry (GEDD-registry, AKTIN) provides an infrastructure for collecting and querying up-to-date medical records in a distributed manner. Within this framework, a benchmark report on cross-institutional comparison using the program R is prepared using routine data of participating hospitals. Currently, 16 emergency departments (EDs) routinely transfer data of 1,200 to approximately 5,000 patients per month to a federated GEDD-registry datawarehouse. Using various packages in the R environment, hospitals receive a monthly visual report on their data among all participating hospitals. Graphical representations are implemented using column diagrams and box plots. Reports currently contain 25 tables and 40 graphs. Benchmark reports are created in R-Studio and exported using Portable Document Format, PDF. Quarterly expert meetings with the heads of participating EDs are currently performed for further improvements. Preparation of external benchmarking reports with R enables a detailed data presentation for participating hospitals and ED managers.

In recent years, increases in the number of patients in emergency departments (ED) have led to continuous work intensification. To handle this problem, the treatment effectiveness has to be maximized. One strategy that may help to optimize workflow is the use of standard operating procedures (SOPs). We investigated the existence of SOPs and subjective effects on treatment in German EDs.We performed an online survey from February 2015 until June 2016. We collected data regarding the existence of SOPs, health care level, medical field, work experience, and education. All professional groups participating in the treatment of patients were requested to take part in the survey.Seventy-five percent of the 589 participants in the survey confirmed the existence of SOPs in their EDs. SOPs were more frequently available in hospitals with higher health care levels. Participants working in EDs without SOPs felt less confident regarding treatment of patients. More than 85% of these participants were in favor of having SOPs. The absence of SOPs was associated with a subjective delay in patient treatment.Most of the EDs had available SOPs. In departments without SOPs, most physicians wanted them to be implemented. SOPs seemed adequate in terms of supporting workflow and satisfaction with patients' treatment.HINTERGRUND: Die in den letzten Jahren steigende Zahl an Patienten in den Notaufnahmen führte zu einer zunehmenden Arbeitsverdichtung. Um dieses Problem zu adressieren, muss u. a. die Behandlungseffektivität gesteigert werden. Eine Strategie ist die Optimierung des Workflows durch die Nutzung von Standard Operating Procedures (SOP). Wir untersuchten die Häufigkeit von SOP und deren subjektive Effekte auf die Patientenbehandlung in deutschen Notaufnahmen.Wir führten von Februar 2015 bis Juni 2016 eine Onlineumfrage in deutschen Notaufnahmen durch. Dazu wurden Daten zum Vorliegen von SOP, dem Versorgungslevel des Krankenhauses sowie dem medizinischen Hintergrund, der Arbeitserfahrung und der Ausbildung des Befragten erhoben. Dabei wurden alle Professionen eingeschlossen, die an der unmittelbaren Patientenbehandlung beteiligt sind.Von den insgesamt 589 Teilnehmern gaben 75 % an, über SOP in der Notaufnahme zu verfügen. Dabei waren SOP häufiger in Krankenhäusern mit höherem Versorgungslevel zu finden. Teilnehmer, die über keine SOP in der Notaufnahme verfügten, fühlten sich unsicherer in der Patientenbehandlung. Mehr als 85 % dieser Befragten wünschten sich SOP. Das Fehlen der SOP war weiterhin mit einer subjektiven Verzögerung der Patientenbehandlung verbunden.Die meisten Notaufnahmen verfügen über SOP. Beim Fehlen von SOP wünschten sich die meisten dort arbeitenden Ärzte deren Implementierung. In diesem Hinblick scheinen SOP ein geeigneter Weg zu sein, den Workflow zu unterstützen und die Zufriedenheit mit der Patientenbehandlung zu verbessern.

Digitalization and standardization of documentation in medicine are increasingly progressing. In the decision of the Federal Joint Committee (G-BA) for a staged system of emergency structures in hospitals and in the report of the expert committee for development in the healthcare system on needs-oriented guidance of healthcare, the actuality of the topic is underlined with concrete naming of future tasks. The section on emergency admission protocols of the German Interdisciplinary Association of Intensive and Emergency Care Medicine (DIVI) has been working for years on this topic and has repeatedly reported on the progress of the topic in clinical emergency care. Standardization and digitalization represent the foundation for health services research spread across locations as well as the possibility for benchmarking. Digitalization makes the secondary use of primary clinical routine data possible. Digitalization decreases redundancies of data transmission by avoiding manual data input in, for example registers.

Gastric carcinogenesis is associated with alterations of microRNAs (miRNAs) and reversal of these alterations may be a crucial element in cancer prevention. Here we evaluate the influence of H. pylori eradication, low-dose aspirin (LDA), non-steroidal anti-inflammatory drugs (NSAIDs) and proton-pump inhibitors (PPI) on modification of inflammatory mucosal miRNAs miR-155 and miR-223 in Helicobacter pylori-infected and non-infected subjects. The study was performed in two parts: 1) interventional study in 20 healthy subjects with and without H. pylori infection or following eradication (each n = 10) where LDA (100 mg) was given daily for 7 days; 2) prospective case-control observational study (n = 188). MiR-155 and miR-223 expression was strongly linked to H. pylori-infection and in short-term view showed a trend for reversal after eradication. Daily LDA as well as regular NSAIDs showed no influence on miRNAs expression both in healthy subjects and patients, while regular PPI intake was associated with lower miR-155 expression in antrum of patients with chronic gastritis independent of density of neutrophils and mononuclear infiltrate. In summary, PPI but not LDA or NSAIDs were associated with modification of inflammatory miRNAs miR-155 and miR-223 in an H. pylori dependent manner. The functional role of inflammatory miR-155 and miR-223 in understanding of H. pylori-related diseases needs further evaluation.

Background Some of the advantages of implementing electronic emergency department information systems (EDIS) are improvements in data availability and simplification of statistical evaluations of emergency department (ED) treatments. However, for multi-center evaluations, standardized documentation is necessary. The AKTIN project (“National Emergency Department Register: Improvement of Health Services Research in Acute Medicine in Germany”) has used the “German Emergency Department Medical Record” (GEDMR) published by the German Interdisciplinary Association of Intensive and Emergency Care as the documentation standard for its national data registry. Methods Until March 2016 the documentation standard in ED was the pen-and-paper version of the GEDMR. In April 2016 we implemented the GEDMR in a timeline-based EDIS. Related to this, we compared the availability of structured treatment information of traumatological patients between pen-and-paper-based and electronic documentation, with special focus on the treatment time. Results All 796 data fields of the 6 modules (basic data, severe trauma, patient surveillance, anesthesia, council, neurology) were adapted for use with the existing EDIS configuration by a physician working regularly in the ED. Electronic implementation increased availability of structured anamnesis and treatment information. However, treatment time was increased in electronic documentation both immediately (2:12 ± 0:04 h; n = 2907) and 6 months after implementation (2:18 ± 0:03 h; n = 4778) compared to the pen-and-paper group (1:43 ± 0:02 h; n = 2523; p

Wilson disease (WD) is a severe hepato-neurologic disorder that affects primarily children and young adults. WD is caused by mutations in ATP7B and subsequent copper overload. However, copper levels alone do not predict severity of the disease. We demonstrate that temporal and spatial distribution of copper in hepatocytes may play an important role in WD pathology. High resolution synchrotron-based x-ray fluorescence imaging in situ indicates that copper does not continuously accumulate in Atp7b−/− hepatocytes, but reaches a limit at 90–300 fmol. The lack of further accumulation is associated with the loss of copper transporter Ctr1 from the plasma membrane and the appearance of copper-loaded lymphocytes and extracellular copper deposits. The WD progression is characterized by changes in subcellular copper localization and transcriptome remodeling. The synchrotron-based x-ray fluorescence imaging and mRNA profiling both point to the key role of nucleus in the initial response to copper overload and suggest time-dependent sequestration of copper in deposits as a protective mechanism. The metabolic pathways, up-regulated in response to copper, show compartmentalization that parallels changes in subcellular copper concentration. In contrast, significant down-regulation of lipid metabolism is observed at all stages of WD irrespective of copper distribution. These observations suggest new stage-specific as well as general biomarkers for WD. The model for the dynamic role of copper in WD is proposed.

Helicobacter pylori, a microaerophilic gram-negative bacterium, colonizes the human stomach. About 50% of the world's population is infected, and this infection is considered as the major risk factor for the development of gastric adenocarcinomas in 1% of infected subjects. Carcinogenesis is characterized by the process of epithelial-to-mesenchymal transition (EMT), in the course of which fully differentiated epithelial cells turn into depolarized and migratory cells. Concomitant disruption of adherence junctions (AJ) is facilitated by growth factors like hepatocyte growth factor 1 (HGF-1), but has been also shown to depend on ectodomain shedding of E-cadherin. The aim of this study was to investigate the impact of infection with H. pylori of NCI-N87 gastric epithelial cells on the shedding of E-cadherin and HGF-receptor c-Met. Our results show that infection with H. pylori provokes shedding of the surface proteins c-Met and E-cadherin. Evidence is provided that ADAM10 contributes to the shedding of c-Met and E-cadherin.

Background & Aims Wilson disease is a severe disorder of copper metabolism caused by mutations in ATP7B, which encodes a copper-transporting adenosine triphosphatase. The disease presents with a variable phenotype that complicates the diagnostic process and treatment. Little is known about the mechanisms that contribute to the different phenotypes of the disease. Methods We analyzed 28 variants of ATP7B from patients with Wilson disease that affected different functional domains; the gene products were expressed using the baculovirus expression system in Sf9 cells. Protein function was analyzed by measuring catalytic activity and copper (64Cu) transport into vesicles. We studied intracellular localization of variants of ATP7B that had measurable transport activities and were tagged with green fluorescent protein in mammalian cells using confocal laser scanning microscopy. Results Properties of ATP7B variants with pathogenic amino-acid substitution varied greatly even if substitutions were in the same functional domain. Some variants had complete loss of catalytic and transport activity, whereas others lost transport activity but retained phosphor-intermediate formation or had partial losses of activity. In mammalian cells, transport-competent variants differed in stability and subcellular localization. Conclusions Variants in ATP7B associated with Wilson disease disrupt the protein's transport activity, result in its mislocalization, and reduce its stability. Single assays are insufficient to accurately predict the effects of ATP7B variants the function of its product and development of Wilson disease. These findings will contribute to our understanding of genotype–phenotype correlation and mechanisms of disease pathogenesis.

Wilson disease (WD) is a severe hepato-neurologic disorder that affects primarily children and young adults. WD is caused by mutations in ATP7B and subsequent copper overload. However, copper levels alone do not predict severity of the disease. We demonstrate that temporal and spatial distribution of copper in hepatocytes may play an important role in WD pathology. High resolution synchrotron-based x-ray fluorescence imaging in situ indicates that copper does not continuously accumulate in Atp7b(-/-) hepatocytes, but reaches a limit at 90-300 fmol. The lack of further accumulation is associated with the loss of copper transporter Ctr1 from the plasma membrane and the appearance of copper-loaded lymphocytes and extracellular copper deposits. The WD progression is characterized by changes in subcellular copper localization and transcriptome remodeling. The synchrotron-based x-ray fluorescence imaging and mRNA profiling both point to the key role of nucleus in the initial response to copper overload and suggest time-dependent sequestration of copper in deposits as a protective mechanism. The metabolic pathways, up-regulated in response to copper, show compartmentalization that parallels changes in subcellular copper concentration. In contrast, significant down-regulation of lipid metabolism is observed at all stages of WD irrespective of copper distribution. These observations suggest new stage-specific as well as general biomarkers for WD. The model for the dynamic role of copper in WD is proposed.