Investigating the correlation between genotypes and hepatic protein expression of CYP2C9, CYP2C19, CYP2D6, and CYP3A5 using postmortem tissue from a Danish population.

The cytochrome (CYP) P450 family of enzymes plays a central role in the metabolism of many drugs. CYP genes are highly polymorphic, which is known to affect protein levels, but for some low frequent CYP genotypes the correlation between genotype and CYP protein expression is less established. In this study, we determined the CYP2C9, CYP2C19, CYP2D6, and CYP3A5 genotypes of 250 Danish individuals included in a postmortem study. For 116 of the individuals, the hepatic CYP protein levels were investigated by a proteomics approach. Overall, we found the postmortem genetic and proteomic data to be in agreement with those of other studies performed on fresh hepatic tissue, showing the usability of postmortem hepatic tissue for this type of investigation. For less investigated genotypes we could corroborate previously found results: 1) statistically significantly lower levels of hepatic CYP2C9 protein in individuals carrying the CYP2C9*3 variant compared to individuals with two wild type (wt) alleles, 2) comparable levels of CYP2C19 in CYP2C19*2/*17 and CYP2C19*1/*2 individuals, 3) reduced CYP2D6 protein levels in heterozygous individuals with the CYP2D6*3, CYP2D6*4, and CYP2D6*5 gene deletion variants, and 4) significantly lower levels of CYP3A5 protein in CYP3A5*3 homozygous individuals compared to individuals that were heterozygous for the CYP3A5*3 allele or homozygous individuals for the wt alleles. In conclusion, the use of postmortem tissue significantly increases the access to human specimens for research purposes, and postmortem proteomics can be used to investigate the link between CYP genotypes and hepatic protein expression. Significance Statement In tissue from a large postmortem cohort (n=250) we determined the CYP2C9, CYP2C19, CYP2D6, and CYP3A5 genotypes. Hepatic CYP protein levels were investigated in 116 individuals using a proteomics approach. For common genotypes, we found results similar to previous knowledge, pointing towards the usability of postmortem tissue. For the less investigated genotypes, we were able to corroborate genotype / protein expression correlations. It is a novel approach to use a large postmortem cohort to investigate genetic / protein expression correlations.

Sex- and Lifestyle-Related Factors are Associated with Altered Hepatic CYP Protein Levels in People Diagnosed with Mental Disorders.

In this study, we used human postmortem tissue to investigate hepatic protein expression levels of cytochrome P450 (CYP) 1A2, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4 by LC-MS/MS in a population of people suffering from mental disorders (n = 171). We report hepatic protein levels of these six CYP isoforms in 171 individuals in total, and define a focused population dataset of 116 individuals after excluding 55 samples due to low microsomal protein per gram of liver (MPPGL) yield. Postmortem decay was most likely the reason for the low MPPGL yield in the 55 samples. In the focused population, we found women to have significantly higher protein levels of CYP3A4 than men in addition to decreased CYP3A4 protein levels among obese individuals. Furthermore, MPPGL was negatively correlated with body mass index (BMI). An increase in CYP1A2 protein levels was observed among smokers, and increased CYP2E1 protein levels were observed among individuals with a history of alcohol abuse. Finally, individuals who received phenobarbital (CYP3A4 inducer) had significantly higher CYP3A4 levels. In conclusion, lifestyle-related factors prevalent among people suffering from mental disorders are associated with altered CYP protein levels, which may alter drug metabolism and affect the efficacy of commonly prescribed drugs. Furthermore, this investigation demonstrates that postmortem hepatic tissue can be used to study how lifestyle and effectors affect hepatic CYP-levels in a large cohort of patients. SIGNIFICANCE STATEMENT: Using a large number of postmortem hepatic tissue specimens (n=116) originating from the autopsy of individuals diagnosed with mental disorders, we were able to show that hepatic CYP-levels were affected by alcohol, smoking, BMI, and sex and that MPPGL was affected by BMI. These lifestyle-related changes may alter drug metabolism and affect the efficacy of commonly prescribed drugs. It is a novel approach to use a large postmortem cohort to investigate how lifestyle and effectors affect hepatic CYP-levels.

Comparing Seizure-Related Death and Suicide in Younger Adults with Epilepsy.

Younger adults with epilepsy have an increased mortality. Some deaths are seizure-related, for example, sudden unexpected death in epilepsy (SUDEP), whereas others, for example, suicide, have multiple causes, including adverse effects of the treatment on mood. In this retrospective population-based study of all Danish persons with epilepsy aged 18 to 49 years during 2007 to 2009 we evaluated the risk of death from seizures and suicide. SUDEP comprised 82.7% of all seizure-related death. Younger adults with epilepsy had an 8.3-fold increased risk of death from seizure-related causes compared with suicide. This underpins the importance of effective seizure control in preventing premature death. ANN NEUROL 2021;90:983-987.

Trends in infant mortality: an evaluation of forensic autopsied infants in Eastern Denmark over 39 years.

BACKGROUND: SIDS is a diagnosis of exclusion applied to the death of an infant < 1 year of age after an extensive post-mortem investigation. From 1980 to 2018, a total of 870 infants have been autopsied at the Section of Forensic Pathology, Department of Forensic Medicine, UCPH, covering East Denmark. In the same period, Danish national guidelines for infant care have been revised to avoid infants dying of SIDS. OBJECTIVE: This study aimed to describe trends in infant autopsies regarding cause and manner of death, gender, age, month of death, sleeping position, and bed-sharing. The trends were compared to the change in national SIDS guidelines during the period of this study. DESIGN: Information from autopsy reports from 1980 to 2018 were collected into 55 categories designed specifically for this study. Data from 7 of these categories were chosen and processed in Excel for basic epidemiological comparison. RESULTS: The trends show that most infants in the study die of natural manner and most predominant causes of death are SIDS, infection, and congenital malformations. A change in national guidelines in 1991 recommending supine- or side sleeping position coincided with a reduction in the overall infant mortality and cases of SIDS. The peak age in the cohort is 90 days, but stratification in decades shows the infants dying younger each decade. Through the study period, the number of infants found dead sleeping in the prone position has declined. Relatively more infants in this cohort have been found dead while bed-sharing, even though the prevalence of these cases has remained largely the same for four decades.

Impact of the Covid-19 lockdown on sexual assault cases in Eastern Denmark - a retrospective clinical forensic study.

To explore if the shutdown of Danish nightlife during the Covid-19 pandemic caused a decrease in the number of clinical forensic examinations of victims of sexual assault in Eastern Denmark. Secondarily, to investigate, if there was a change in criminological characteristics, e.g. scene and time of crime, relation to the perpetrator and the proportion of possible drug-facilitated sexual assaults. 130 case files from clinical forensic examinations of individuals of alleged sexual assault in the period 1st of April to 30th of June in both 2019 and 2020 were included. 67 and 63 examinations were performed in 2019 and 2020, respectively. 125 cases were female and five were male. Approximately 70% were 15-25 years of age. Pre- and post-lockdown victim profiles were similar regarding assailant relation, location of crime and time of assault. Voluntary intake of alcohol prior to the assault was registered with 46.3% in 2019 and 62% in 2020. The ratio of possible drug-facilitated sexual assault (DFSA) was approximately 50% each year. The lockdown did not seem to change the overall number of examinations or the demographic and criminological characteristics of the sexual assault victims. No decrease in cases of possible DFSA was found despite the lockdown of nightlife venues.

Sudden unexpected death in epilepsy in persons younger than 50 years: A retrospective nationwide cohort study in Denmark.

OBJECTIVE: Persons with epilepsy have an increased mortality including a high risk of sudden unexplained death (SUD), also referred to as sudden unexpected death in epilepsy (SUDEP). We aimed to evaluate the risk of SUDEP in comparison to other causes of death and the risk of SUD in persons with and without epilepsy. METHODS: We undertook a retrospective population-based cohort study of all Danish citizens with and without epilepsy aged 1-49 years during 2007-2009. All deaths in the population were evaluated, and all cases of SUD identified. Primary causes of death in persons with epilepsy were evaluated independently by three neurologists and one neuropediatrician, using the unified SUDEP criteria. RESULTS: The three most frequent causes of death in persons with epilepsy were cancer (2.38 per 1000 person-years), SUDEP (1.65 per 1000 person-years), and pneumonia (1.09 per 1000 person-years) compared with cancer (.17 per 1000 person-years), accident-related deaths (.14 per 1000 person-years), and cardiovascular disease (.09 per 1000 person-years) in persons without epilepsy. Considering definite, definite plus, and probable cases, the SUDEP incidence was .27 per 1000 person-years (95% confidence interval [CI] = .11-.64) in children aged 1-17 years and 1.21 per 1000 person-years (95% CI = .96-1.51) in adults aged 18-49 years. Adjusted for age and sex, persons with epilepsy younger than 50 years had a 10.8-fold (95% CI = 9.97-11.64, p < .0001) increased all-cause mortality and a 34.4-fold (95% CI = 23.57-50.28, p < .0001) increased risk of SUD compared with persons without epilepsy. SUDEP accounted for 23.3% of all SUD. SIGNIFICANCE: This nationwide study of all deaths in persons with epilepsy younger than 50 years found a lower SUDEP risk in children compared with adults, and that epilepsy was a major risk factor for SUD in the background population. This underlines the importance of addressing risk factors for SUDEP to prevent premature death.

The usefulness of a trauma probability of survival model for forensic life-threatening danger assessments.

Clinical forensic medical examinations constitute an increasing proportion of our institution's tasks, and, concomitantly, the authorities are now requesting forensic life-threatening danger assessments based on our examinations. The aim of this retrospective study was to assess if a probability of survival (PS) trauma score could be useful for these forensic life-threatening danger assessments and to identify a cut-off PS score as a supporting tool for the forensic practice of assessing life-threatening danger. We compared a forensic database and a trauma database and identified 161 individuals (aged 15 years or older) who had both a forensic life-threatening danger assessment and a PS score. The life-threatening danger assessments comprised the following statements: was not in life-threatening danger (NLD); could have been in life-threatening danger (CLD); or was in life-threatening danger (LD). The inclusion period was 2012-2016. A statistically significant difference was found in the PS scores between NLD, CLD and LD (chi-square test: p < 0.0001). The usefulness of the PS score for categorizing life-threatening danger assessments was determined by a receiver-operator characteristic (ROC) curve. The area under the curve was 0.76 (95% CI, 0.69 to 0.84) and the ROC curve revealed that a cut-off PS score of 95.8 would appropriately identify LD. Therefore, a PS score below 95.8 would indicate life-threatening danger. We propose a further exploration of how the evidence-based PS score, including a cut-off value, might be implemented in clinical forensic medical statements to add to the scientific strength of these statements.

Life-threatening danger assessments of penetrating injuries in Eastern Danish clinical forensic medicine.

Clinical forensic assessments of injuries' life-threatening danger may have an impact on the legal aftermath following a violent assault. The pursuit of evidence-based guidelines should ensure a user-independent and reproducible forensic practice. However, does it? The aim of this study was to evaluate the forensic life-threatening danger assessments after a protocol implementation in 2016. The evaluation concerned usability and reproducibility of the protocol, and its influence on assessment severity. We analyzed the level of inter- and intra-rater agreement using 169 blinded, prior-protocol cases that were reassessed by two forensic specialists. We compared assessment made the year before and after protocol implementation (n = 262), and the forensic specialists' reassessments with the prior-protocol cases' original assessments (n = 169). Whether to make an assessment, the levels of agreement varied between weak agreement (inter-rater, Κ = 0.43; assessor 1, Κ = 0.57) and strong agreement (assessor 2, Κ = 0.90). Regarding severity, the levels of agreement varied between strong agreement (inter-rater, Κ = 0.87; assessor 1: Κ = 0.90) and almost perfect agreement (assessor 2: Κ = 0.94). The assessments were statistically significant redistributed after the implementation (chi-square test: p < 0.0001). The proportion of cases assessed as having not been in life-threatening danger increased from 9 to 43%, and moderate severity assessments decreased from 55 to 23%. Of the moderate severity assessments, 55% were reassessed as having not been in life-threatening danger. The protocol ensured independent and reproducible assessments when the forensic specialists agreed on making one. The protocol resulted in less severe assessments. Future studies should examine the reliability of the protocol and its consequences for legal aftermaths.

Genetic investigations of 100 inherited cardiac disease-related genes in deceased individuals with schizophrenia.

Cardiac diseases and sudden cardiac death (SCD) are more prevalent in individuals diagnosed with schizophrenia compared to the general population, with especially coronary artery disease (CAD) as the major cardiovascular cause of death. Antipsychotic medications, genetics, and lifestyle factors may contribute to the increased SCD in individuals with schizophrenia. The role of antipsychotic medications and lifestyle factors have been widely investigated, while the genetic predisposition to inherited cardiac diseases in schizophrenia is poorly understood. In this study, we examined 100 genes associated with inherited cardiomyopathies and cardiac channelopathies in 97 deceased individuals diagnosed with schizophrenia for the prevalence of genetic variants associated with SCD. The deceased individuals had various causes of death and were included in the SURVIVE project, a prospective, autopsy-based study of mentally ill individuals in Denmark. This is the first study of multiple inherited cardiac disease-related genes in deceased individuals with diagnosed schizophrenia to shed light on the genetic predisposition to SCD in individuals with schizophrenia. We found no evidence for an overrepresentation of rare variants with high penetrance in inherited cardiac diseases, following the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG) consensus guidelines. However, we found that the deceased individuals had a statistically significantly increased polygenic burden caused by variants in the investigated heart genes compared to the general population. This indicates that common variants with smaller effects in heart genes may play a role in schizophrenia.

Sudden cardiac death among persons with diabetes aged 1-49 years: a 10-year nationwide study of 14 294 deaths in Denmark.

AIMS: The aim of this study was to compare nationwide incidence rate (IR) of sudden cardiac death (SCD) in persons aged 1-49 years with and without diabetes mellitus (DM). METHODS AND RESULTS: The study population consisted of all persons in Denmark aged 1-49 years in 2000-09, which equals 27.1 million person-years. All 14 294 deaths in the 10-year period were included. By using the highly descriptive Danish death certificates, 1698 cases of sudden and unexpected death were identified. Through review of autopsy reports, discharge summaries, and the Danish registries, we identified 1363 cases of SCD. The Danish Register of Medicinal Product Statistics was used to identify persons with type 1 DM and type 2 DM. Among the 14 294 decedents, there were 669 with DM, of which 118 suffered SCD (9% of all SCD), making SCD the leading cause of death among young persons with DM. Among those aged 1-35 years, the IR of SCD-DM was 21.9 per 100 000 person-years compared to 2.6 per 100 000 person-years among persons without DM [IR ratio 8.6, 95% confidence interval (CI) 5.8-28.6]. Within the age range 36-49 years, the IR among persons with DM was 119.8 per 100 000 person-years compared to 19.7 per 100 000 person-years among persons without DM (IR ratio 6.1, 95% CI 4.7-7.8). CONCLUSION: We found that young persons with DM aged 1-35 years had >8-fold higher SCD IR compared to young persons without DM. Our study highlights the need for early cardiovascular risk monitoring and assessment in young persons with DM.

Undiagnosed disease in decedents with and without schizophrenia: an autopsy-based case-control study.

BACKGROUND: Individuals with schizophrenia (SCZ) suffer from higher morbidity and mortality than the general population. Some of this is due to concurrent somatic disease. AIMS: To identify and compare the somatic comorbidities antemortem (AM) and postmortem (PM) in autopsied decedents with SCZ and with no mental illness (NMI), using the Charlson Comorbidity Index (CCI). MATERIALS: A total of 106 autopsied decedents with SCZ and 156 decedents with NMI were included. AM diagnoses were sampled from Danish national health registers. PM diagnoses were included from forensic autopsies. Negative binomial regression models were used to test associations between age, sex, SCZ diagnosis, manner of death and the CCI score both AM and PM. RESULTS: The CCI score increased significantly from AM to PM for both decedents with SCZ and NMI (SCZ, AM 0.669 vs PM 1.208, p = 0.008) (NMI, AM 0.519 vs PM 1.218, p = 0.000). Regression analysis showed that age was significantly associated with the CCI score both AM and PM (AM, OR 1.044 [1.029-1.060] p ≤ 0.001) (PM, OR 1.028 [1.015-1.041], p ≤ 0.001). A diagnosis of SCZ was correlated with the CCI score AM but not PM (AM, OR 1.880 [1.207-2.928], p = 0.005) (PM, OR 1.170 [0.828-1.654], p = 0.374). CONCLUSION: The autopsies revealed undiagnosed diseases in both decedents with SCZ and NMI. The diseases were primarily of cardiovascular, pulmonary and gastrointestinal origin and may have contributed to premature mortality. Age was the only covariate significantly associated with the CCI score both AM and PM; the SCZ diagnosis was positively correlated to the CCI score before death.

Psychiatric disorders in a population of deceased drug users.

PURPOSE: To estimate the prevalence of psychiatric morbidity and dual diagnosis in a population of decedents with positive drug toxicology and evaluate changes over time between 2001-2002 and 2011-2012. MATERIALS AND METHODS: A total of 520 autopsied drug users with positive toxicology were included in the study from 2001 to 2002 and from 2011 to 2012. Materials included autopsy reports, toxicological screening during autopsy and data from the Danish national health registers, including psychiatric diagnoses from psychiatric hospitals and ambulatory functions, dispensed prescription use from pharmacies and registered treatment for drug use disorders. RESULTS: In 2001-2002, 63.3% of the decedents had only positive toxicology, 22.5% also had psychiatric morbidity, and 14.2% had a dual diagnosis. In 2011-2012, 56.4% had only positive toxicology, 26.1% also had psychiatric morbidity, and 17.5% had a dual diagnosis. None of the changes were significant. Decedents with only positive toxicology became older at time of death over time; decedents with psychiatric morbidity and a dual diagnosis did not. The prevalence of nonprescribed psychotropic medication, methadone and benzodiazepines increased. CONCLUSION: Decedents with psychiatric morbidity and dual diagnosis did not increase their lifespan over a 10-year period. Decedents with only positive toxicology increasingly consumed nonprescribed psychotropic medication and may have suffered from undiagnosed psychiatric disorders. The prevalence of prescribed and nonprescribed benzodiazepines and methadone increased and may have contributed to premature mortality.

Differential Methylation in the GSTT1 Regulatory Region in Sudden Unexplained Death and Sudden Unexpected Death in Epilepsy.

Sudden cardiac death (SCD) is a diagnostic challenge in forensic medicine. In a relatively large proportion of the SCDs, the deaths remain unexplained after autopsy. This challenge is likely caused by unknown disease mechanisms. Changes in DNA methylation have been associated with several heart diseases, but the role of DNA methylation in SCD is unknown. In this study, we investigated DNA methylation in two SCD subtypes, sudden unexplained death (SUD) and sudden unexpected death in epilepsy (SUDEP). We assessed DNA methylation of more than 850,000 positions in cardiac tissue from nine SUD and 14 SUDEP cases using the Illumina Infinium MethylationEPIC BeadChip. In total, six differently methylated regions (DMRs) between the SUD and SUDEP cases were identified. The DMRs were located in proximity to or overlapping genes encoding proteins that are a part of the glutathione S-transferase (GST) superfamily. Whole genome sequencing (WGS) showed that the DNA methylation alterations were not caused by genetic changes, while whole transcriptome sequencing (WTS) showed that DNA methylation was associated with expression levels of the GSTT1 gene. In conclusion, our results indicate that cardiac DNA methylation is similar in SUD and SUDEP, but with regional differential methylation in proximity to GST genes.

Undiagnosed acromegaly as an underlying cause of sudden death.

We report a case of sudden death in a 31-year-old male diagnosed at autopsy with clinical undiagnosed acromegaly. The purpose of this report is to underline the importance of health professionals reacting to phenotypic acromegaly, such as acral enlargement and/or unexplained hypertension, including a range of severe comorbidities, to avoid a fatal outcome. Recent studies have shown that the increased mortality seen in acromegaly patients can be reversed with modern treatment aimed at normalizing GH and IGF-I levels. One year before death, the presented case was diagnosed with hypertension, but was otherwise described as healthy. The forensic autopsy, including post-mortem CT, showed phenotypic facial and body characteristics for acromegaly, general visceromegaly, and a pituitary tumor. The cause of death was heart failure due to end-stage acromegalic cardiopathy. Because the disease is slowly progressive, the individual himself, and the people close to him, might not have considered the acromegaly-related facial changes as abnormal.

Enlargement of the human adrenal zona fasciculata and chronic psychiatric illness - an autopsy-based study.

Severe mental illness (SMI) is associated with a reduced life expectancy of up to 20 years. One possible contributor to this fact is dysregulation of the hypothalamus-pituitary-adrenal (HPA)-axis. Looking at the morphology of effector organs, such as the adrenal glands themselves, could reveal insights into organ function and response to possible HPA-dysregulation. This forensic autopsy-based study investigated if there were any morphological changes in adrenal glands between decedents who had previously been submitted to a psychiatric hospital with a diagnosis of schizophrenia (n = 34), bipolar (n = 5), or depressive disorder (n = 20), any other psychiatric diagnosis (n = 36) compared with decedents who had no previous psychiatric admission (n = 40). Length of admissions to psychiatric wards and admission in the 180 days preceding death was included in regression as proxy variables for severity of illness. On the macroscopic level, we found no difference in gland weight or volume. On the microscopic level, we found a 25% increase in cross-sectional area of the zona fasciculata (ZF) in decedents who had a diagnosis of schizophrenia compared with controls (p = 0.033). Other diagnosis groups did not differ from controls. Total admission length was positively correlated with area of the ZF.Lay SummaryPeople with a severe mental disorder may be in a constant state of increased stress, which is harmful. This study looked at the adrenal gland, which produces stress hormones, to see if they were different in deceased persons who had suffered from a severe mental illness. We found that the part of the adrenal gland that produces stress hormones is larger in deceased patients who suffered from schizophrenia, but not other types of psychiatric illnesses, compared to deceased persons with no history of psychiatric illness.

Reliability of police reports when assessing health information at the forensic post-mortem examination-using schizophrenia as a model.

Autopsies continue to be the most reliable source of mortality statistics; however, more and more death certificates are based on the post-mortem external examination (PME) alone. Forensic PMEs differ from clinical PMEs, because the forensic pathologist usually has no preceding knowledge of the health of the decedent and must rely on information from authorities in the form of the police report. It is useful at the forensic PME to know whether the decedent suffered from a mental illness; however, it is unknown how valid such a diagnosis is, when based upon information in the police report alone. This study compared tentative diagnoses of schizophrenia from 500 forensic PMEs with a reference database based on the Danish National Patient Registry. We found that 19.3% of schizophrenia cases were missed, and 9.1 % of identified cases were false positives. Overall, 11.4% of all assessments were incorrect. Subgroup analysis showed that marital status as 'single' and the finding of illegal substances at the scene were predictors for both correctly identified and overlooked schizophrenia cases. The most reliable source of information was the decedent's general practitioner, whereas friends and neighbors were the most unreliable. Future studies should be aware of the risk of assigning a wrong diagnosis and use as many sources of information as possible. Taking the decedent's social history and observations about the scene into account may add to the diagnostic accuracy.

Targeted molecular genetic testing in young sudden cardiac death victims from Western Denmark.

Sudden unexpected death in the young continues to be an important unsolved challenge. A significant proportion of the deaths are suspected to be caused by inherited cardiac diseases and are referred to as sudden cardiac deaths (SCD). We performed targeted molecular testing of 70 deceased individuals under 40 years of age that after forensic autopsy were suspected to have died of SCD. The individuals were previously genetically investigated using smaller numbers of genes associated with specific cardiac diseases. In our previous studies, seven (10%) individuals had pathogenic or likely pathogenic variants according to the 2015 ACMG guidelines. In order to investigate the value of expanding the panel to 100 genes associated with cardiac diseases, we histopathologically re-examined the 70 suspected SCD cases and grouped them according to phenotypes into suspected cardiomyopathy (the cardiomyopathy group), left ventricular hypertrophy (the hypertrophy group) and structural normal hearts (the SUD group). DNA was captured with the Haloplex target enrichment system and sequenced using an Illumina MiSeq. We found that 11 (16%) individuals harboured pathogenic or likely pathogenic variants. In the cardiomyopathy, hypertrophy and SUD groups, 22%, 6% and 17% of the individuals, respectively, harboured pathogenic or likely pathogenic variants. Our findings show that testing of a broad panel of genes associated with cardiac diseases identify potential pathogenic variants of cardiac diseases in a significant proportion of SCD cases, and this may have important implications in family screening to prevent future deaths.

Targeted exon sequencing in deceased schizophrenia patients in Denmark.

Schizophrenia patients have higher mortality rates and lower life expectancy than the general population. However, forensic investigations of their deaths often fail to determine the cause of death, hindering prevention. As schizophrenia is a highly heritable condition and given recent advances in our understanding of the genetics of schizophrenia, it is now possible to investigate how genetic factors may contribute to mortality. We made use of findings from genome-wide association studies (GWAS) to design a targeted panel (PsychPlex) for sequencing of exons of 451 genes near index single nucleotide polymorphisms (SNPs) identified with GWAS. We sequenced the DNA of 95 deceased schizophrenia patients included in SURVIVE, a prospective, autopsy-based study of mentally ill persons in Denmark. We compared the allele frequencies of 1039 SNPs in these cases with the frequencies of 2000 Danes without psychiatric diseases and calculated their deleteriousness (CADD) scores. For 81 SNPs highly associated with schizophrenia and CADD scores above 15, expression profiles in the Genotype-Tissue Expression (GTEx) Project indicated that these variants were in exons, whose expressions are increased in several types of brain tissues, particularly in the cerebellum. Molecular pathway analysis indicated the involvement of 163 different pathways. As for rare SNP variants, most variants were scored as either benign or likely benign with an average of 17 variants of unknown significance per individual and no pathogenic variant. Our results highlight the potential of DNA sequencing of an exon panel to discover genetic factors that may be involved in the development of schizophrenia.

Treatment Status and Use of Psychoactive Substances in Deceased Drug Users.

BACKGROUND: Studies on drug use are limited by the study populations available, which usually only include drug users in treatment settings. Therefore, the knowledge base is limited on drug users not entering treatment for drug use disorder (DUD). Using registers from departments of forensic medicine enables research on decedents with DUD, irrespective of treatment status. OBJECTIVES: The aim of this study is to characterize and compare drug users not receiving treatment and drug users receiving treatment, in relation to cause of death, toxicological findings, and use of nonprescribed medication. METHODS: Retrospective register-based study on deceased drug users with supplemental data from the Registry of Drug Abusers Undergoing Treatment and the Register of Medicinal Product Statistics in 2 observation periods: 2001-2002 and 2011-2012. RESULTS: Two-thirds of the population were not receiving treatment at the time of death in both observation periods. Drug users receiving treatment were more likely to die from accidental poisonings than drug users not receiving treatment. There was no difference in mean age at the time of death between the 2 groups, and both groups were older in the second observation period. There was no difference in toxicological findings according to treatment status and the 2 groups did not differ in the presence of nonprescribed medication found in the blood at the time of death. DISCUSSION/CONCLUSIONS: The proportion of drug users that received treatment prior to death has not increased, and deceased drug users are mostly not in treatment for their drug use at the time of death.

Spontaneous Subarachnoid Haemorrhage as a Cause of Out-of-Hospital Death.

BACKGROUND: There are no recent studies on the incidence rate of out-of-hospital death due to spontaneous subarachnoid haemorrhage (SAH). The primary aim of this study was to determine how often SAH was the cause of out-of-hospital death. The secondary aim was to determine if decedents had contacted any health care services within the last 72 h prior to the time of death. METHODS: This was a retrospective cohort study. The reports of all autopsies carried out at the Department of Forensic Medicine in the Capital Region of Denmark in a ten-year period were read. Police records and Emergency Medical Services (EMS) telephone records were searched for health care contacts within the last 72 h prior to the time of death. Descriptive statistics were used, and to analyse the incidence rates for trend Poisson regression was used. RESULTS: In total, 6,903 decedents underwent autopsy. Out-of-hospital SAH was the cause of death in 58 decedents, resulting in an average incidence rate of 0.34 per 100.000 persons per year. No significant change in the incidence rate over time was found (p = 0.52). No EMS data were available for eleven decedents in the first part of the study period. Of the remaining 47 decedents, 2 (3.5%, 95% CI: 0.4-11.9) had called the EMS, and in regards to 27 of the 58 decedents, the police records contained information on health care system contacts. Five (8.6%, 95% CI: 2.9-18.9) patients had contacted a general practitioner and three (5.2%, 95% CI: 1.1-14.4) patients had been admitted to hospital but were discharged again within 72 h prior to their death. CONCLUSION: The incidence rate of out-of-hospital death from SAH was 0.34 per 100.000 persons per year and remained stable across the years 2009-2018. Several patients had sought medical attention shortly before their death, emphasizing the vital importance of recognizing the early symptoms of SAH.