Single cell transcriptomics reveals distinct transcriptional responses to oxycodone and buprenorphine by iPSC-derived brain organoids from patients with opioid use disorder.

The opioid epidemic represents a national crisis. Oxycodone is one of the most prescribed opioid medications in the United States, whereas buprenorphine is currently the most prescribed medication for opioid use disorder (OUD) pharmacotherapy. Given the extensive use of prescription opioids and the global opioid epidemic, it is essential to understand how opioids modulate brain cell type function at the single-cell level. We performed single nucleus RNA-seq (snRNA-seq) using iPSC-derived forebrain organoids from three male OUD subjects in response to oxycodone, buprenorphine, or vehicle for seven days. We utilized the snRNA-seq data to identify differentially expressed genes following drug treatment using the Seurat integrative analysis pipeline. We utilized iPSC-derived forebrain organoids and single-cell sequencing technology as an unbiased tool to study cell-type-specific and drug-specific transcriptional responses. After quality control filtering, we analyzed 25787 cells and identified sixteen clusters using unsupervised clustering analysis. Our results reveal distinct transcriptional responses to oxycodone and buprenorphine by iPSC-derived brain organoids from patients with OUD. Specifically, buprenorphine displayed a significant influence on transcription regulation in glial cells. However, oxycodone induced type I interferon signaling in many cell types, including neural cells in brain organoids. Finally, we demonstrate that oxycodone, but not buprenorphine activated STAT1 and induced the type I interferon signaling in patients with OUD. These data suggest that elevation of STAT1 expression associated with OUD might play a role in transcriptional regulation in response to oxycodone. In summary, our results provide novel mechanistic insight into drug action at single-cell resolution.

Genome-wide association study for circulating FGF21 in patients with alcohol use disorder: Molecular links between the SNHG16 locus and catecholamine metabolism.

OBJECTIVE: Alcohol consumption can increase circulating levels of fibroblast growth factor 21 (FGF21). The effects of FGF21 in the central nervous system are associated with the regulation of catecholamines, neurotransmitters that play a crucial role in reward pathways. This study aims to identify genetic variants associated with FGF21 levels and evaluate their functional role in alcohol use disorder (AUD). METHODS: We performed a genome-wide association study (GWAS) using DNA samples from 442 AUD subjects recruited from the Mayo Clinic Center for the Individualized Treatment of Alcoholism Study. Plasma FGF21 levels were measured using Olink proximity extension immunoassays. Alcohol consumption at time of entry into the study was measured using the self-reported timeline followback method. Functional genomic studies were performed using HepG2 cells and induced pluripotent stem cell (iPSC)-derived brain organoids. RESULTS: Plasma FGF21 levels were positively correlated with recent alcohol consumption and gamma-glutamyl transferase levels, a commonly used marker for heavy alcohol use. One variant, rs9914222, located 5' of SNHG16 on chromosome 17 was associated with plasma FGF21 levels (p = 4.60E-09). This variant was also associated with AUD risk (ß: -3.23; p:0.0004). The rs9914222 SNP is an eQTL for SNHG16 in several brain regions, i.e., the variant genotype was associated with decreased expression of SNHG16. The variant genotype for the rs9914222 SNP was also associated with higher plasma FGF21 levels. Knockdown of SNHG16 in HepG2 cells resulted in increased FGF21 concentrations and decreased expression and enzyme activity for COMT, an enzyme that plays a key role in catecholamine metabolism. Finally, we demonstrated that ethanol significantly induced FGF21, dopamine, norepinephrine, and epinephrine concentrations in iPSC-derived brain organoids. CONCLUSIONS: GWAS for FGF21 revealed a SNHG16 genetic variant associated with FGF21 levels which are associated with recent alcohol consumption. Our data suggest that SNHG16 can regulate FGF21 concentrations and decrease COMT expression and enzyme activity which, in turn, have implications for the regulation of catecholamines. (The ClinicalTrials.gov Identifier: NCT00662571).

Dietary and plasma carotenoids are positively associated with alpha diversity in the fecal microbiota of pregnant women.

Because microbes use carotenoids as an antioxidant for protection, dietary carotenoids could be associated with gut microbiota composition. We aimed to determine associations among reported carotenoid intake, plasma carotenoid concentrations, and fecal bacterial communities in pregnant women. Pregnant women (n = 27) were enrolled in a two-arm study designed to assess feasibility of biospecimen collection and delivery of a practical nutrition intervention. Plasma and fecal samples were collected and women were surveyed with a 24-hr dietary checklist and recalls. Plasma carotenoids were analyzed by HPLC using photodiode array detection. Fecal bacteria were analyzed by 16S rRNA DNA sequencing. Results presented are cross-sectional from the 36-week gestational study visit combined across both study arms due to lack of significant differences between intervention and usual care groups (n = 23 women with complete data). Recent intake of carotenoid-containing foods included carrots, sweet potatoes, mangos, apricots, and/or bell peppers for 48% of women; oranges/orange juice (17%); egg (39%); tomato/tomato-based sauces (52%); fruits (83%); and vegetables (65%). Average plasma carotenoid concentrations were 6.4 µg/dL α-carotene (AC), 17.7 µg/dL ß-carotene (BC), 11.4 µg/dL cryptoxanthin, 39.0 µg/dL trans-lycopene, and 29.8 µg/dL zeaxanthin and lutein. AC and BC concentrations were higher in women who recently consumed foods high in carotenoids. CR concentrations were higher in women who consumed oranges/orange juice. Microbiota α-diversity positively correlated with AC and BC. Microbiota ß-diversity differed significantly across reported intake of carotenoid containing foods and plasma concentrations of AC. This may reflect an effect of high fiber or improved overall dietary quality, rather than a specific effect of carotenoids. PRACTICAL APPLICATION: Little is known about the association between the gut microbiome and specific dietary microconstituents, such as carotenoids, especially during pregnancy. This research demonstrates that a carotenoid-rich diet during pregnancy supports a diverse microbiota, which could be one mechanism by which carotenoids promote health.

Substance use disorders and their effect on the psychiatric and justice systems.

BACKGROUND AND OBJECTIVES: A high proportion of persons in institutionalized settings such as the criminal justice system and psychiatric hospitals have substance use disorders (SUDs). We explored the association between substance use, demographics, and criminal justice involvement in a population of patients placed on involuntary 72-h holds in a psychiatric facility. METHODS: We retrospectively identified patients aged 18 through 57 years who had been placed on 72-h holds during an acute psychiatric hospitalization during a 1-year period. Data were analyzed with standard descriptive statistics, and data collection was reviewed by 2 randomly assigned psychiatrists. RESULTS: We identified 336 patients placed on 72-h holds during an acute psychiatric stay. Of these, more than two-thirds (68.5%; n = 230) had an SUD. Compared with patients not using substances, those with SUDs were significantly more likely to be younger (p = .003), male (p = .005), and unmarried (p < .001) and to have criminal justice involvement before (p < .001) and after hospitalization (p < .001). The rate of unemployment was similarly high in both users (67.4%) and nonusers (69.2%). DISCUSSION AND CONCLUSIONS: Most patients on involuntary psychiatric holds have comorbid SUDs. These patients are more likely to have interacted with the criminal justice system and less likely to have social support in the form of marriage. Unemployment was common among all patients. SCIENTIFIC SIGNIFICANCE: When SUDs are not treated by the criminal justice or mental health system, rehospitalization and criminal recidivism may result. (Am J Addict 2018;27:574-577).

Alcohol reduction in the first trimester is unrelated to smoking, patient or pregnancy characteristics.

INTRODUCTION: Studies show alcohol-preferring mice reduce their alcohol intake during pregnancy; this study questions if the same is true for humans. The current investigation compares women's pre-pregnancy and first trimester alcohol consumption, examines if women with problem drinking diminish their alcohol intake during pregnancy, and determines if prenatal alcohol reduction is associated with characteristics of pregnancy, patients or smoking. METHODS: 126 participants in weeks 1-12 of pregnancy, recruited from Obstetric and Family Practices, completed a survey during their initial prenatal visit including two gender-specific AUDITs (Alcohol Use Disorders Identification Tests) querying current and pre-pregnancy alcohol use. AUDIT-C (AUDIT items 1-3) scores measuring pre-pregnancy and first trimester alcohol consumption were compared, analyzed and tested using general linear model repeated. A p ≤ 0.05 was accepted as significant. RESULTS: Most participants were multiparous, Caucasian high school graduates experiencing nausea and vomiting. Pre-pregnancy alcohol use was significantly (p = 0.019, Fisher's exact) higher among women seeing obstetricians. Pre-pregnancy AUDIT-C scores (m (mean) = 2.22, sd (standard deviation) = 2.19) were significantly higher (p < 0.001) than first trimester scores (m = 0.143, sd = 0.532). Among 49 with pre-pregnancy AUDIT-C scores ≥ 3, 45/49 (92%) reduced their alcohol use to zero during the first trimester. Age, race, education, marital status, parity, nausea and vomiting, gestational age and smoking were non-factors in score reduction. CONCLUSIONS: Women reported reducing their alcohol consumption during pregnancy, including those screening positive for pre-pregnancy problem drinking. First trimester alcohol reduction cannot be accounted for by smoking, patient or pregnancy characteristics; public health initiatives, psychological factors and hormonal mechanisms may be implicated.

Mixed mania associated with cessation of breastfeeding.

BACKGROUND: This case chronicles the unique presentation of psychotic mixed mania in a female 5 months after parturition and 1 week following breastfeeding discontinuation, highlighting a rarely recognized mania risk factor that is temporally delayed from parturition: breastfeeding discontinuation. CASE PRESENTATION: A 25-year-old G1P1 female with a past psychiatric history of a depressive episode in adolescence presented to the Emergency Department with her 5-month-old daughter, fiancée, and family 1 week after breastfeeding cessation. She endorsed sleep-deprived energy enhancement, unfulfilled goal-oriented productivity, hyper-talkativeness, hyper-sexuality and increased nicotine use. Concurrent depressive symptoms included hopelessness, worthlessness, poor concentration, lack of appetite, and ego-dystonic intrusive thoughts that she may kill herself or her child. She exhibited pressured speech, affective lability, expansiveness, distractibility, and tangential, grandiose, delusional self-referential content. Transient thoughts of self-harm and harm to her child were not associated with intent. Her family history was significant for a deceased mother who had bipolar I disorder. The patient was hospitalized for 5 days and diagnosed with bipolar disorder, type I, current episode manic with psychotic features with a mixed-feature specifier. Olanzapine and lithium were initiated and the patient's acute episode of mania resolved prior to discharge. CONCLUSIONS: This case extends the limited literature on mania following weaning and highlights the role of rapid serum dopamine rise following breastfeeding cessation in mania.

The role of gabapentin in the management of alcohol withdrawal and dependence.

OBJECTIVE: To review the literature evaluating gabapentin for alcohol withdrawal and dependence. DATA SOURCES: A literature search of MEDLINE (1966 to end of March 2015) and PubMed was performed using the terms alcohol, gabapentin, withdrawal, and dependence. Additional references were identified from a review of literature citations. STUDY SELECTION AND DATA EXTRACTION: English-language prospective studies evaluating gabapentin for alcohol withdrawal and dependence were evaluated. DATA SYNTHESIS: A total of 10 publications utilizing gabapentin in alcohol withdrawal (n = 5) and alcohol dependence (n = 5) were included in this review. Limited data suggest that gabapentin can provide benefit in managing mild alcohol withdrawal syndrome. There were 5 reported or suspected seizures in the withdrawal studies, suggesting that additional safety data are necessary before gabapentin monotherapy can be routinely considered. Sleep and mood/anxiety-related outcomes were positively influenced by gabapentin, which may result in long-term benefits if continued beyond the withdrawal period for the treatment of alcohol dependence. Studies evaluating gabapentin for alcohol dependence demonstrated dose-dependent benefits for complete abstinence, rates of no heavy drinking, and cravings. Gabapentin used to treat alcohol dependence was well tolerated with no severe adverse reactions reported in the extant literature. CONCLUSION: Gabapentin may have a role in the treatment of mild alcohol withdrawal, but future studies should focus on adequate dosing strategies. Gabapentin should be considered for the treatment of alcohol dependence when barriers prevent the use of traditional agents. Additional studies should be conducted to further validate findings from the research conducted to date, but the current literature is promising for gabapentin in the treatment of alcohol dependence.

Restoration of ovarian activity and pregnancy after transplantation of cryopreserved ovarian tissue: a review of 60 cases of reimplantation.

Aggressive chemotherapy/radiotherapy and bone marrow transplantation can cure >90% of girls and young women affected by disorders requiring such treatment. However, the ovaries are very sensitive to cytotoxic drugs, especially to alkylating agents. Several options are currently available to preserve fertility in cancer patients. The present review reports the results of 60 orthotopic reimplantations of cryopreserved ovarian tissue performed by three teams, as well as 24 live births reported in the literature to date. Restoration of ovarian activity occurred in almost all cases in the three series. Among the 60 patients, eleven conceived and six of those had already delivered twelve healthy babies. In the future, we are looking to: 1) improve freezing techniques; and 2) enhance the "vascular bed" before reimplantation to increase pregnancy rates. On the other hand, cryopreservation of ovarian tissue may be combined with removal, via puncture, of small antral follicles, making it possible to freeze both ovarian tissue and isolated immature oocytes.

Short-term risk after initial observation for chest pain.

Few studies have evaluated the necessity of immediate stress testing after observation for chest pain. The purpose of this study was to assess the safety of outpatient stress testing after discharge from a chest pain unit. We hypothesized that discharge from a chest pain unit before stress testing is associated with a low rate of short-term adverse outcomes. This was a retrospective chart review of managed care patients discharged from the chest pain unit before the performance of stress testing. Records were reviewed for the occurrence of adverse cardiac outcomes before an outpatient stress test up to 60 days post-discharge. Primary outcomes were defined as death or myocardial infarction, and secondary outcomes as readmission for chest pain evaluation, unstable angina, or congestive heart failure. Three hundred forty-four patients were identified. One hundred sixty-six patients had either a recent prior stress test (17) or an outpatient test (149) performed within 60 days of discharge. During that time, 2 patients (0.6%) had a fatal out-of-hospital cardiac event, and there were 27 subsequent chest pain visits to the Emergency Department by 24 patients (7.0%). Nine patients (2.6%) were admitted to the hospital and 10 (2.9%) were readmitted to the observation unit for chest pain. We conclude that patients who have negative serial electrocardiograms and enzyme testing in a chest pain unit are at low risk for short-term cardiac events. Appropriately selected patients may be discharged for subsequent outpatient testing.

Anxiety sensitivity as a specific and unique marker of anxious symptoms in youth psychiatric inpatients.

We addressed several questions regarding the relation of anxiety sensitivity to anxious symptoms among 47 youth psychiatric inpatients (18 boys, 29 girls), ages 9-17 (M = 14.23, SD = 1.89). Participants completed measures of anxiety sensitivity, anxious and depressive symptoms, trait anxiety, and positive and negative affect; chart diagnoses were available. Consistent with hypotheses, we found that (a) anxiety sensitivity was associated with anxious symptoms, even controlling for trait anxiety and depressive symptoms; and (b) anxiety sensitivity displayed symptom specificity to anxious versus depressive symptoms (i.e., was associated with anxiety controlling for depression but not with depression controlling for anxiety). Furthermore, regarding factors of anxiety sensitivity, we obtained mixed support for our prediction that phrenophobia would be associated with both depression and anxiety, whereas fear of physical arousal would be associated with anxiety but not depression. Implications for the construct validity of anxiety sensitivity were discussed.

Cognitive factors differentiating attention deficit-hyperactivity disorder with and without a comorbid mood disorder.

Mood disorders and attention deficit-hyperactivity disorder (ADHD) co-occur in 20-30% of children and adolescents diagnosed in both epidemiological and clinical studies, but little information is available regarding cognitive factors that may be relevant to the expression of co-occurring mood disorders and ADHD. This study examined whether ADHD with and without a comorbid mood disorder could be differentiated on the basis of cognitive factors associated with prominent theories of depression. Children meeting diagnostic criteria for ADHD (n = 14) or ADHD and a comorbid mood disorder (n = 27) were assessed on a variety of cognitive indices. Children in the comorbid group reported more negative views of themselves and a more depressogenic attributional style. Cognitive disturbances associated with A. T. Beck's (1967) cognitive model and attributional style theories of depression differentiate ADHD children with significant mood pathology.