Childhood maltreatment is a major risk factor for depression and may contribute to sex differences in depression prevalence. We examined sex-specific associations between childhood maltreatment and depression and estimated the proportion of depression cases attributable to maltreatment subtypes. We analyzed baseline data from 159,045 participants (49.4% female; aged 19-72) in the German National Cohort (NAKO). Childhood maltreatment was assessed via the Childhood Trauma Screener; depression via self-reported physician's diagnosis and MINI classification (lifetime) and the PHQ-9 (current). Associations, including sex interactions, were modeled using binary logistic regressions. Disparity decomposition and sex-stratified population attributable fractions quantified the contribution of childhood maltreatment to depression. Childhood maltreatment was associated with increased odds of lifetime (ORphysician's diagnosis=2.45 [2.38,2.53]; ORMINI=2.30 [2.18,2.43]) and current depression (OR=2.90 [2.79,3.02], all padj<0.001). Sex interactions were observed for physician's diagnosis: physical abuse (padj=0.024) and neglect (padj<0.001) had stronger associations in females (ORphysical abuse=2.74 [2.59,2.90]; ORphysical neglect=1.36 [1.28,1.44]) than males (ORphysical abuse=2.36 [2.21,2.52]; ORphysical neglect=1.08 [1.00,1.16]), whereas sexual abuse (padj=0.003) showed stronger associations in males (OR=3.23 [2.91,3.57]) than females (OR=2.61 [2.48,2.75]). Overall, childhood maltreatment accounted for 21.2-26.2% of lifetime and 33.4% of current depression. Population attributable fractions were higher in females than males for lifetime (24.5-28.5% vs. 16.0-20.9%) and current depression (36.1% vs. 28.4%). Emotional subtypes contributed the highest population attributable fractions (up to 10.2%). Childhood maltreatment contributed 19.2-22.4% to the association between sex and depression. Childhood maltreatment accounts for a substantial proportion of depression in both sexes, with stronger overall associations in females. Sex-specific prevention may help reduce depression prevalence.

Substance use disorders (SUDs) are complex psychiatric conditions influenced by genetic and environmental factors. DNA methylation (DNAm), a dynamic and context-dependent epigenetic mechanism, can offer insight into regulatory variation associated with SUDs, particularly when examined in brain regions central to addiction-related neurobiology. In this systematic review, we compiled findings from 20 methylome-wide association studies (MWASs) of SUDs in postmortem human brain tissue, encompassing alcohol (nstudy=9), opioids (nstudy=4), cocaine (nstudy=3), nicotine (nstudy=2), methamphetamine (nstudy=1), and any illicit drug (nstudy=1). Studies varied in brain regions analyzed, methylation platforms used, and sample characteristics. Most focused on methylation at cytosines followed by guanine (i.e., mCGs). Despite methodological heterogeneity and limited overlap of findings within individual substances, consistent biological themes emerged across substances, including differential methylation in genes related to neurodevelopment, inflammatory response, signaling, and transcriptional regulation. Cross-substance comparisons revealed 525 genes implicated across independent results from at least two substances, suggesting shared methylomic signals that may reflect common neurobiological pathways, downstream adaptations to exposure, or unmeasured polysubstance use. Exploratory drug repurposing analyses based on genes overlapping three or more substances identified a small number of existing compounds with potential relevance for treating polysubstance use. This review synthesizes current results to highlight both the promise and challenges of SUD MWASs in human brain, including limited sample size, generalizability, cell-type specificity, and lack of functional validation. Future research should prioritize multi-omic integration, larger and more diverse cohorts, and advanced technologies to enhance mechanistic understanding and translational potential.

Borderline personality disorder (BPD) is a severe mental health condition influenced by environmental risk factors (for example, interpersonal trauma) and genetic factors. We conducted the largest genome-wide association study (GWAS) meta-analysis of BPD so far, with a discovery sample of 12,339 cases and 1,041,717 controls, and a replication study of 685 cases and 107,750 controls (all participants of European ancestry). We identified 11 independent associated genomic loci and 9 risk genes in gene-based analyses. We observed a single-nucleotide polymorphism heritability of 17.3% and derived polygenic scores (PGS) that predicted 4.6% of the phenotypic variance in BPD on the liability scale. BPD showed the strongest positive genetic correlations with GWAS of post-traumatic stress disorder, depression, attention deficit hyperactivity disorder, antisocial behavior, and measures of suicide and self-harm. Phenome-wide analyses in Vanderbilt University Medical Center Biobank and UK Biobank using BPD-PGS confirmed these associations and also identified associations with other medical conditions, including obstructive pulmonary disease and diabetes. These analyses highlight BPD as a polygenic disorder, with the genetic risk showing substantial overlap with psychiatric and physical health conditions.

The ability of environmental cues to trigger alcohol-seeking behaviours is thought to facilitate problematic alcohol use. Individuals' tendency to attribute incentive salience to cues may increase the risk of addiction. We sought to study the relationship between incentive salience and alcohol addiction using non-preferring rats to model the heterogeneity of human alcohol consumption, investigating both males and females. Adult rats were subjected to the alcohol deprivation effect (ADE) paradigm, where they were given voluntary access to different alcohol solutions with repeated interruptions by deprivation and reintroduction phases over a protracted period (five Alcohol Deprivation Cycles). Before each Alcohol Deprivation Cycle, rats were tested in the Pavlovian Conditioned Approach (PCA) paradigm, which quantifies the individual salience toward a conditional cue and the reward, thus allowing us to trace the process of attributing incentive salience to reward cues. During the final Alcohol Deprivation Cycle (ADE5), animals were tested for compulsive-like behaviour using quinine taste adulteration. We investigated sex differences in drinking behaviour and PCA performance. We observed thatb females drank significantly more alcohol than males and displayed more sign-tracking (ST) behaviour in the PCA, whereas males showed goal-tracking (GT) behaviour. Furthermore, we found that high drinkers exhibited more ST behaviour. The initial PCA phenotype was correlated with later alcohol consumption. Our findings indicate a complex relationship between incentive salience and alcohol addiction and emphasize the importance of considering both sexes in preclinical research.

We conducted the largest genome-wide meta-analysis of borderline personality disorder (BPD) to date, with a discovery sample of 12,339 cases and 1,041,717 controls, and a replication study of 685 cases and 107,750 controls (all participants of European ancestry). We identified 11 independent associated genomic loci, and nine risk genes in the gene-based analysis. We observed a single-nucleotide polymorphism (SNP) heritability of 17.3% and derived polygenic scores (PGS) predicted 4.6% of the phenotypic variance in BPD on the liability scale. BPD showed the strongest positive genetic correlations with GWAS of posttraumatic stress disorder, depression, attention deficit hyperactivity disorder, antisocial behavior, and measures of suicide and self-harm. Phenome-wide association analyses using BPD-PGS confirmed these associations and additionally revealed associations with general medical conditions including obstructive pulmonary disease and diabetes. The present analyses highlight BPD as a polygenic disorder, with the genetic risk showing substantial overlap with psychiatric and physical health conditions.

Despite extensive research on DNA methylation (DNAm) signatures associated with alcohol use disorder (AUD), findings are often inconsistent and not replicated. We conducted a large-scale meta-analysis of epigenome-wide association studies (EWAS) to identify reliable, reproducible epigenetic markers of AUD. Seven cohorts, comprising 3,775 individuals (1,325 with AUD), contributed to this meta-analysis within the framework of the Psychiatric Genomics Consortium Substance Use Disorders Epigenetics Working Group. Downstream analyses included the identification of differentially methylated regions, overrepresentation analyses, and the construction of a methylation risk score (MRS). We identified 118 significant CpG sites associated with AUD, with the strongest association found at cg24889777 ( p =5.12×10 -17 ) in the long non-coding RNA LOC100505942. CpG sites were enriched for pathways related to GTPase signaling and transmembrane transporter activity, as well as EWAS signals of alcohol consumption. The MRS explained 10.44% of variance in heavy drinking in an independent cohort (N=2,534, AUC=0.657). This large-scale meta-analysis offers key insights into the epigenetic mechanisms of AUD and lays the groundwork for future research on methylation risk scores for the diagnosis, prognosis, and treatment in AUD.

BACKGROUND:Although the association between smoking and depression is well-established, the underlying mechanisms and contextual factors remain insufficiently understood. We examined the association between smoking and depression, including detailed dose-response and timing-related relationships, using baseline data from a large population-based cohort, the German National Cohort (NAKO). METHODS:The analysis comprised 173,890 participants (19-72 years, 50.21% female). Lifetime and current depression were assessed via self-reported physician's diagnosis, the Major Depressive Disorder module of the MINI International Neuropsychiatric Interview (MINI), and the depression scale of the Patient Health Questionnaire (PHQ-9). Smoking behavior was assessed using self-reported smoking status, age at initiation, cigarettes per day, and time since smoking cessation. Associations between smoking and depression measures were analyzed using regression models adjusted for sex, age, age², education, Body Mass Index, and alcohol consumption. RESULTS:Lifetime depression was more prevalent among individuals who currently or formerly smoked compared to those who never smoked. Currently smoking individuals also reported most current depressive symptoms, followed by formerly smoking individuals and those who never smoked. A dose-response relationship was observed, with more cigarettes per day being associated with more current depressive symptoms. Later age at smoking initiation was associated with later depression onset. Time since smoking cessation was positively associated with time since last depressive episode and negatively with current depressive symptoms. CONCLUSIONS:Our findings support an association between smoking and depression. Robust dose-response relationships were found, with higher cigarette consumption associated with more severe depressive symptoms, and longer time since cessation linked to lower depression levels. These results highlight smoking as a meaningful and modifiable contributor to current and lifetime depression, suggesting that quitting smoking or reducing cigarette consumption may benefit mental health. Early prevention of smoking initiation, along with integrated approaches that combine smoking cessation support with mental health care, may help reduce both smoking rates and depression burden.

BACKGROUND:Impulsivity is a key feature of bipolar disorder (BD) associated with various negative outcomes. Recent use of ecological momentary assessment (EMA) has allowed for nuanced examination of the mechanisms of mood and impulsivity dysregulation. However, few existing studies have used an ecological momentary assessment of impulsivity in multiplex families with BD and examined its associations with mood. OBJECTIVE:Using EMA, this study investigated the concurrent and predictive relationships between impulsivity and mood. METHODS:Multiplex family members with BD (BDF, n = 8), unaffected family members (FC, n = 6), individuals with BD not from families (BDC, n = 8) and healthy controls (HC, n = 8), completed daily EMA surveys about mood and impulsivity for 6-12 weeks. Mixed-effects regression concurrent and lagged models were employed to analyze the relationship between impulsivity and mood. RESULTS:The BDF (Diff = -31.70, p = 0.001) and BDC (Diff = -25.74, p = 0.007) groups had a significantly lower mean in mood scores compared to the HC group but not compared to the FC group. There were no significant differences in the mean impulsivity scores between the groups. Time-lagged analyses revealed a significant negative association between prior impulsivity and mood at the next assessment independent of diagnosis (OR=0.939, p = 0.002). However, the opposite relationship between prior mood and impulsivity was not significant (OR=0.996, p = 0.135). CONCLUSIONS:These results contribute to the understanding of the complex interactions between BD, the genetic load of the disorder, impulsivity and mood. Furthermore, these findings indicate the potential benefits of addressing impulsivity as a means to improve mood outcomes at an early stage.

Psychotic-like experiences (PLEs) have been identified as risk factors for mental health issues and behavioral problems including violence. While cross-sectional studies suggest an association between PLEs and violent behavior in adolescents, their longitudinal relationship remains unexamined. This study aims to examine the temporal association between PLEs and violent behavior in adolescents. PLEs and violent behavior were assessed using data from self-report surveys conducted from 2011 to 2019 in a Tokyo junior and senior high school (grades 7–12). The study included 1685 participants aged 12–18 surveyed annually for up to 6 years. Random intercept cross-lagged panel models (RI-CLPMs) were used to examine between-person and within-person associations among study variables, with analyses stratified by gender. Results showed a bidirectional relationship between PLEs and violent behavior on both the between-person ( β = 0.23, p < 0.001) and within-person levels ( β = 0.07–0.25, p < 0.05). This relationship was significant for PLEs and violence towards objects (between-person: β = 0.22, p < 0.001; within-person: β = 0.07–0.32, p < 0.05), but not for PLEs and interpersonal violence. When analyzed by gender, these associations were significant in girls but not in boys. The findings suggested that PLEs may have a bidirectional relationship with violent behavior (especially violence towards objects), particularly in girls, indicating potential gender-specific pathways in this association. Further research should explore the underlying mechanisms of this bidirectional relationship, with a focus on gender-specific factors.

Electroconvulsive therapy (ECT) is an effective antidepressant treatment. The mechanisms behind the therapeutic effect are not fully understood, and reliable biomarkers for response are needed. Epigenetic modifications, such as DNA methylation (DNAm), can reflect both genetic and environmental impacts; they may shed light on the mechanisms behind treatment effects and they have the potential to inform response prediction. We performed an epigenome-wide association study (EWAS) in peripheral blood from patients before and after ECT in a Norwegian cohort (n = 65). The methylation levels of 12 differentially methylated CpG positions (DMPs) and 18 differentially methylated regions (DMRs) were significantly associated with percent clinical response. In addition, 29 DMPs and 23 DMRs were significantly associated with remission (Montgomery and Åsberg Depression Rating Scale MADRS < 10 post treatment). Two DMRs were also significantly associated with percent response at baseline and four DMRs were significantly associated with remission at baseline (FDR < 0.05). We did not identify any longitudinal (pre-post) changes in DNAm. We further performed the first meta-analysis (n = 99) between ECT cohorts, combining this Norwegian cohort and a German ECT cohort (n = 34). Seven of the DMRs found to be associated with response in the meta-analyses were previously identified in the Norwegian or the German cohort (FDR < 0.05). Methylation risk scores (MS) calculated using DMPs associated with ECT in the Norwegian cohort showed promising association with response to ECT in the German cohort (p = 0.06). Finally, we found increased neutrophil to lymphocyte ratios, calculated from estimated cell proportions, to be associated with remission (p < 0.003) in the Norwegian cohort.

BACKGROUND:Alcohol use disorder (AUD) is associated with increased mortality and morbidity risk. A reason for this could be accelerated biological aging, which is strongly influenced by disease processes such as inflammation. As recent studies of AUD show changes in DNA methylation and gene expression in neuroinflammation-related pathways in the brain, biological aging represents a potentially important construct for understanding the adverse effects of substance use disorders. Epigenetic clocks have shown accelerated aging in blood samples from individuals with AUD. However, no systematic evaluation of biological age measures in AUD across different tissues and brain regions has been undertaken. METHODS:As markers of biological aging (BioAge markers), we assessed Levine's and Horvath's epigenetic clocks, DNA methylation telomere length (DNAmTL), telomere length (TL), and mitochondrial DNA copy number (mtDNAcn) in postmortem brain samples from Brodmann Area 9 (BA9), caudate nucleus, and ventral striatum (N = 63-94), and in whole blood samples (N = 179) of individuals with and without AUD. To evaluate the association between AUD status and BioAge markers, we performed linear regression analyses while adjusting for covariates. RESULTS:The majority of BioAge markers were significantly associated with chronological age in all samples. Levine's epigenetic clock and DNAmTL were indicative of accelerated biological aging in AUD in BA9 and whole blood samples, while Horvath's showed the opposite effect in BA9. No significant association of AUD with TL and mtDNAcn was detected. Measured TL and DNAmTL showed only small correlations in blood and none in brain. CONCLUSIONS:The present study is the first to simultaneously investigate epigenetic clocks, telomere length, and mtDNAcn in postmortem brain and whole blood samples in individuals with AUD. We found evidence for accelerated biological aging in AUD in blood and brain, as measured by Levine's epigenetic clock, and DNAmTL. Additional studies of different tissues from the same individuals are needed to draw valid conclusions about the congruence of biological aging in blood and brain.

Background:Mobile devices for remote monitoring are inevitable tools to support treatment and patient care, especially in recurrent diseases such as major depressive disorder. The aim of this study was to learn if machine learning (ML) models based on longitudinal speech data are helpful in predicting momentary depression severity. Data analyses were based on a dataset including 30 inpatients during an acute depressive episode receiving sleep deprivation therapy in stationary care, an intervention inducing a rapid change in depressive symptoms in a relatively short period of time. Using an ambulatory assessment approach, we captured speech samples and assessed concomitant depression severity via self-report questionnaire over the course of 3 weeks (before, during, and after therapy). We extracted 89 speech features from the speech samples using the Extended Geneva Minimalistic Acoustic Parameter Set from the Open-Source Speech and Music Interpretation by Large-Space Extraction (audEERING) toolkit and the additional parameter speech rate. Objective:We aimed to understand if a multiparameter ML approach would significantly improve the prediction compared to previous statistical analyses, and, in addition, which mechanism for splitting training and test data was most successful, especially focusing on the idea of personalized prediction. Methods:To do so, we trained and evaluated a set of >500 ML pipelines including random forest, linear regression, support vector regression, and Extreme Gradient Boosting regression models and tested them on 5 different train-test split scenarios: a group 5-fold nested cross-validation at the subject level, a leave-one-subject-out approach, a chronological split, an odd-even split, and a random split. Results:In the 5-fold cross-validation, the leave-one-subject-out, and the chronological split approaches, none of the models were statistically different from random chance. The other two approaches produced significant results for at least one of the models tested, with similar performance. In total, the superior model was an Extreme Gradient Boosting in the odd-even split approach (R²=0.339, mean absolute error=0.38; both P<.001), indicating that 33.9% of the variance in depression severity could be predicted by the speech features. Conclusions:Overall, our analyses highlight that ML fails to predict depression scores of unseen patients, but prediction performance increased strongly compared to our previous analyses with multilevel models. We conclude that future personalized ML models might improve prediction performance even more, leading to better patient management and care.

Laboratory-based studies have shown that psychological stress caused by response to various stressors triggers acute changes in the cardiovascular system. A better understanding is needed of the emerging evidence on temporal associations between psychological stress and cardiovascular responses in natural settings. This study examined the association of psychological stress and heart rate variability (HRV) in daily life, at high resolution over 2 weeks, taking the effect of physical activity into account. Participants (n = 34) completed ecological momentary assessments (EMA) 6 times per day, reporting levels of perceived stress, low-arousal negative affect (LNA), and high-arousal negative affect. Chest-mounted heart-rate monitors were worn to assess HRV. Multilevel models were used to examine the association between psychological stress levels and preceding/subsequent HRV. Reduced time domain HRV measures (mean and standard deviation of R-wave to R-wave intervals) during the prior hour predicted higher levels of perceived stress. Frequency domain HRV measures higher low to high frequency (LF/HF) and lower HF to total power (HF nu) ratios during the preceding 10 min predicted higher perceived stress levels, suggesting the dominance of sympathetic nervous system activity. EMA reports of higher perceived stress levels were associated with reduced time domain HRV measures during the following 10 min. On the other hand, higher LNA were related to increased HRV measures, such as lower LF/HF and higher HF nu during the following hour. The dynamic associations observed may have therapeutic implications for 'just-in-time' interventions in the management of daily stress and cardiovascular health.

BACKGROUND:Digital phenotyping and monitoring tools are the most promising approaches to automatically detect upcoming depressive episodes. Especially, linguistic style has been seen as a potential behavioral marker of depression, as cross-sectional studies showed, for example, less frequent use of positive emotion words, intensified use of negative emotion words, and more self-references in patients with depression compared to healthy controls. However, longitudinal studies are sparse and therefore it remains unclear whether within-person fluctuations in depression severity are associated with individuals' linguistic style. METHODS:To capture affective states and concomitant speech samples longitudinally, we used an ambulatory assessment approach sampling multiple times a day via smartphones in patients diagnosed with depressive disorder undergoing sleep deprivation therapy. This intervention promises a rapid change of affective symptoms within a short period of time, assuring sufficient variability in depressive symptoms. We extracted word categories from the transcribed speech samples using the Linguistic Inquiry and Word Count. RESULTS:Our analyses revealed that more pleasant affective momentary states (lower reported depression severity, lower negative affective state, higher positive affective state, (positive) valence, energetic arousal and calmness) are mirrored in the use of less negative emotion words and more positive emotion words. CONCLUSION:We conclude that a patient's linguistic style, especially the use of positive and negative emotion words, is associated with self-reported affective states and thus is a promising feature for speech-based automated monitoring and prediction of upcoming episodes, ultimately leading to better patient care.

BACKGROUND:Adolescents are vulnerable to mental health problems, and this vulnerability may be enhanced in situations such as the present COVID-19 pandemic. Online mental health literacy (MHL) education may help adolescents maintain/improve their mental health, especially in situations where face-to-face education is difficult. AIMS:To evaluate the effects of a teacher-led "online Short MHL Program (o-SMHLP)" delivered online to grade 10 students in their classrooms. METHODS:Students (age 15-16) were randomly assigned to an o-SMHLP group (n = 115 (3 classes)) or a control group (n = 155 (4 classes)) at the class level. The program consisted of a 20-minute session which included an animated video. The students completed a self-report questionnaire pre- and post-intervention assessing outcomes including: "Knowledge about mental health/illnesses", "Recognition of necessity to seek help", "Intention to seek help", and "Unwillingness to socialize with people having mental illness". Mixed effects modeling was employed for analyses. RESULTS:All outcomes were significantly improved in the intervention group compared to the control group post-intervention, except for "intention to seek help". CONCLUSIONS:The present study shows the effectiveness of an online MHL intervention while identifying the need for the development of effective online programs targeting adolescents' "intention to seek help".

IntroductionParents and guardians (hereafter caregivers) of teenagers need high levels of mental health literacy (MHL) to manage mental health problems arising in teenagers in their care. Previous studies assessing MHL levels in caregivers of teenagers have reported mixed results, making it difficult to clearly estimate caregiver MHL levels. This study aimed to investigate MHL levels in Japanese caregivers of regular teenagers.MethodsResponses from caregivers (n = 1,397) of students entering junior and senior high schools to a self-administered online questionnaire were analyzed. The questionnaire assessed (a) knowledge about mental health/illnesses and (b) attitudes towards mental health problems in teens in their care (e.g., recognition of depression as a medical illness and intention to engage in helping behaviors).ResultsThe average proportion of correct answers to the knowledge questions (n = 7) was 55.4%; about one tenth (9.2%) of caregivers correctly answered only one or none of the questions. Few caregivers correctly answered about the life-time prevalence of any mental illnesses (46.1%) and appropriate sleep duration for teenagers’ health (16.5%). The proportions of caregivers who had the intention to listen to the teen in their care, consult another person, and seek professional medical help if the teen suffered from depression were 99.5%, 91.5% and 72.7%, respectively.ConclusionsMany teenagers’ caregivers appeared to be willing to help the teens in their care if they were suffering from mental health problems. However, there was much room for improvement in knowledge on mental health/illnesses and intention to seek help from medical professionals. Efforts toward better education should be made.

Abstract Background Having no or few confidants is found to be associated with more severe mental health problems and a higher prevalence of depression in adults, but research examining this association in adolescents is scarce. Social relationships may be particularly critical during adolescence, as it is an important developmental period during which vulnerability to mental health problems increases. The present study examined the relationship between having no or few confidants and anxiety/depressive symptoms in adolescents. Methods Cross-sectional self-report survey targeting 7–12th grade students (age range: 12–18) was conducted in public junior and senior high schools in Mie and Kochi, Japan. Data from 17,829 students (49.7% boys) were analyzed. Associations between anxiety/depressive symptoms (12-item General Health Questionnaire; score range: 0–12) and the number of confidants (None, 1–3, or ≥ 4) were examined using multilevel regression analyses. The analyses were stratified by gender and school level (junior/senior high), and adjusted for experiences of being physically abused and bullied and the interactions of these experiences with the number of confidants. Results Having no or 1–3 confidants was associated with more anxiety/depressive symptoms, compared to having ≥ 4 confidants (p < 0.001) in all stratified groups. Having no confidants was associated with more anxiety/depressive symptoms than having 1–3 confidants (p < 0.001); in senior high boys, no difference was observed between having no confidants and having 1–3 confidants. In addition, in senior high boys, victims of bullying who have confidants reported significantly less anxiety/depressive symptoms than the victims who have no confidants (p < 0.01). Conclusions Adolescents who had no or few confidants had more anxiety/depressive symptoms. Attention needs to be paid to better identify these adolescents, and avenues to support them need to be established.

Pharmacogenomics aims to use the genetic information of an individual to personalize drug prescribing. There is evidence that pharmacogenomic testing before prescription may prevent adverse drug reactions, increase efficacy, and reduce cost of treatment. CYP2D6 is a key pharmacogene of relevance to multiple therapeutic areas. Indeed, there are prescribing guidelines available for medications based on CYP2D6 enzyme activity as deduced from CYP2D6 genetic data. The Agena MassARRAY system is a cost-effective method of detecting genetic variation that has been clinically applied to other genes. However, its clinical application to CYP2D6 has to date been limited by weaknesses such as the inability to determine which haplotype was present in more than one copy for individuals with more than two copies of the CYP2D6 gene. We report application of a new protocol for CYP2D6 haplotype phasing of data generated from the Agena MassARRAY system. For samples with more than two copies of the CYP2D6 gene for which the prior consensus data specified which one was present in more than one copy, our protocol was able to conduct CYP2D6 haplotype phasing resulting in 100% concordance with the prior data. In addition, for three reference samples known to have more than two copies of CYP2D6 but for which the exact number of CYP2D6 genes was unknown, our protocol was able to resolve the number for two out of the three of these, and estimate the likely number for the third. Finally, we demonstrate that our method is applicable to CYP2D6 hybrid tandem configurations.

Objectives Problematic sexual behavior (PSB) is defined by recurrent sexual behaviors that are difficult to control, causing social and functional impairments. PSB can co-occur with reward deficiency syndrome (RDS), but this relationship remains unclear. RDS has been associated with the 10/10 genotype of 3 ′ variable number tandem repeat (VNTR) in the dopamine transporter gene SLC6A3, which is implicated in the reward pathway. This study investigates the genetic relationship between PSB and RDS, testing their association with SLC6A3 3′ VNTR genotype. Methods PSB patients from addiction treatment facilities (n=454), and comparison participants (with PSB=82; without PSB, n=888) were recruited. PSB was measured by the Sexual Addiction Screening Test-Revised (SAST-R) Core and RDS was measured using a composite variable from a custom test battery. DNA was collected from saliva and buccal swabs. Genotyping was performed using polymerase chain reaction (PCR), and regression analyses were conducted to investigate the association of SLC6A3 3′ VNTR genotype with PSB and RDS. Results The 10/10 genotype of SLC6A3 3′ VNTR was associated with RDS in a combined analysis of all groups, and with PSB only in comparison participants. In patients, rare SLC6A3 3′ VNTR genotypes (3-, 6-, 8-, 11-repeat alleles) were associated with PSB. No genotype showed relationships to RDS in only PSB patients. Conclusions The link of the 10/10 genotype to RDS indicates that PSB could be a manifestation of RDS in non-clinical populations; rare genotypes might be associated with clinical forms of PSB, together with comorbid psychopathology. ### Competing Interest Statement While Dr Carnes was previously a Board member of the American Foundation for Addiction Research, he is no longer a member. Moreover, neither the Foundation nor any other of the funders played any role in study design, or in data analysis or interpretation thereof. All other authors declare no conflicts of interest. ### Funding Statement The work herein was supported by an Alberta Centennial Addiction and Mental Health Research Chair and transitional funding (to KJA), Canada Foundation for Innovation (CFI), John R. Evans Leaders Fund (JELF) grant (32147 - Pharmacogenetic translational biomarker discovery), Alberta Innovation and Advanced Education Small Equipment Grants Program (to KJA), and a research grant and philanthropic support from the American Foundation for Addiction Research (to KJA). A Fulbright-Canada-Palix Foundation grant (to PC) assisted with his contributions to study design, project management, and collaborative working. SJ was supported by an Alberta Innovates Postdoctoral Recruitment Fellowship. ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The Quorum Review Institutional Review Board (Seattle, Washington; Protocol Number: 2016-001) and the University of Alberta Research Ethics Board (Protocol: Pro00066552) approved the study. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present study are available upon reasonable request to the authors.

BackgroundThe use of mobile devices to continuously monitor objectively extracted parameters of depressive symptomatology is seen as an important step in the understanding and prevention of upcoming depressive episodes. Speech features such as pitch variability, speech pauses, and speech rate are promising indicators, but empirical evidence is limited, given the variability of study designs. ObjectivePrevious research studies have found different speech patterns when comparing single speech recordings between patients and healthy controls, but only a few studies have used repeated assessments to compare depressive and nondepressive episodes within the same patient. To our knowledge, no study has used a series of measurements within patients with depression (eg, intensive longitudinal data) to model the dynamic ebb and flow of subjectively reported depression and concomitant speech samples. However, such data are indispensable for detecting and ultimately preventing upcoming episodes. MethodsIn this study, we captured voice samples and momentary affect ratings over the course of 3 weeks in a sample of patients (N=30) with an acute depressive episode receiving stationary care. Patients underwent sleep deprivation therapy, a chronotherapeutic intervention that can rapidly improve depression symptomatology. We hypothesized that within-person variability in depressive and affective momentary states would be reflected in the following 3 speech features: pitch variability, speech pauses, and speech rate. We parametrized them using the extended Geneva Minimalistic Acoustic Parameter Set (eGeMAPS) from open-source Speech and Music Interpretation by Large-Space Extraction (openSMILE; audEERING GmbH) and extracted them from a transcript. We analyzed the speech features along with self-reported momentary affect ratings, using multilevel linear regression analysis. We analyzed an average of 32 (SD 19.83) assessments per patient. ResultsAnalyses revealed that pitch variability, speech pauses, and speech rate were associated with depression severity, positive affect, valence, and energetic arousal; furthermore, speech pauses and speech rate were associated with negative affect, and speech pauses were additionally associated with calmness. Specifically, pitch variability was negatively associated with improved momentary states (ie, lower pitch variability was linked to lower depression severity as well as higher positive affect, valence, and energetic arousal). Speech pauses were negatively associated with improved momentary states, whereas speech rate was positively associated with improved momentary states. ConclusionsPitch variability, speech pauses, and speech rate are promising features for the development of clinical prediction technologies to improve patient care as well as timely diagnosis and monitoring of treatment response. Our research is a step forward on the path to developing an automated depression monitoring system, facilitating individually tailored treatments and increased patient empowerment.