Subregion and sex differences in ethanol activation of cholinergic and glutamatergic cells in the mesopontine tegmentum.

Ethanol engages cholinergic signaling and elicits endogenous acetylcholine release. Acetylcholine input to the midbrain originates from the mesopontine tegmentum (MPT), which is composed of the laterodorsal tegmentum (LDT) and the pedunculopontine tegmental nucleus (PPN). We investigated the effect of acute and chronic ethanol administration on cholinergic and glutamatergic neuron activation in the PPN and LDT in male and female mice. We show that ethanol activates neurons of the PPN and not the LDT in male mice. Chronic 15 daily injections of 2 g/kg ethanol induced Fos expression in cholinergic and glutamatergic PPN neurons in male mice, whereas ethanol did not increase cholinergic and glutamatergic neuronal activation in the LDT. A single acute 4 g/kg injection, but not a single 2 g/kg injection, induced cholinergic neuron activation in the male PPN but not the LDT. In contrast, acute or chronic ethanol at either dose or duration had no effect on the activation of cholinergic or glutamatergic neurons in the MPT of female mice. Female mice had higher baseline level of activation in cholinergic neurons compared with males. We also found a population of co-labeled cholinergic and glutamatergic neurons in the PPN and LDT which were highly active in the saline- and ethanol-treated groups in both sexes. These findings illustrate the complex differential effects of ethanol across dose, time point, MPT subregion and sex.

Subregion and sex differences in ethanol activation of cholinergic and glutamatergic cells in the mesopontine tegmentum.

Ethanol engages cholinergic signaling and elicits endogenous acetylcholine release. Acetylcholine input to the midbrain originates from the mesopontine tegmentum (MPT), which is composed of the laterodorsal tegmentum (LDT) and the pedunculopontine tegmental nucleus (PPN). We investigated the effect of acute and chronic ethanol administration on cholinergic and glutamatergic neuron activation in the PPN and LDT in male and female mice. We show that ethanol selectively activates neurons of the PPN and not the LDT in male mice. Acute 4.0 g/kg and chronic 15 daily injections of 2.0 g/kg i.p. ethanol induced Fos expression in cholinergic and glutamatergic PPN neurons in male mice, whereas cholinergic and glutamatergic neurons of the LDT were unresponsive. In contrast, acute or chronic ethanol at either dose or duration had no effect on the activation of cholinergic or glutamatergic neurons in the MPT of female mice. Female mice had higher level of baseline activation in cholinergic neurons compared with males. We also found a population of co-labeled cholinergic and glutamatergic neurons in the PPN and LDT which were highly active in the saline- and ethanol-treated groups in both sexes. These findings illustrate the complex differential effects of ethanol across dose, time point, MPT subregion and sex.

Back-translating GWAS findings to animal models reveals a role for Hgfac and Slc39a8 in alcohol and nicotine consumption.

Alcohol and tobacco are the most commonly used addictive substances, with high comorbidity rates between alcohol use disorder and tobacco use disorder. Risk for alcohol and nicotine addiction is highly heritable, and they share common genetic factors. A GWAS in over 1 million individuals has revealed 566 genetic variants in 406 loci associated with multiple stages of alcohol and tobacco use. Three novel genes-SLC39A8, GRK4 and HGFAC-within loci associated with altered alcoholic drinks per week (ADW) or cigarettes per day (CPD) were selected to further study their role in alcohol and tobacco use disorder. The role of these genes was assessed using the two-bottle choice addiction paradigm in transgenic mice for each of the genes. We found significant decreases in chronic alcohol consumption and preference in female Hgfac knockout (KO) mice, and decreased nicotine preference in male Hgfac KO compared with wild-type (WT) mice. Additionally, male Slc39a8 hypomorph mice showed greater overall nicotine preference compared with WT mice, while no differences were detected for Grk4 KO mice in alcohol or nicotine consumption and preference in either sex. Thus, this study implicates Hgfac and Slc39a8 in alcohol and tobacco use in a sex-specific manner.

Assessment of automated craving across substances and across cultures: stability-analysis of the Craving Automated Scale (CAS).

BACKGROUND: The transition from hedonic to compulsive use in Substance Use Disorders (SUD) is a critical point in SUD progression and hence relevant for assessment and treatment. To measure the habitual patterns of substance consumption, the Craving Automated Scales (CAS) for alcohol (CAS-A), substances (CAS-S) and cigarette smoking (CAS-CS) were developed and introduced to different countries. In this study, we aimed to investigate the structural stability of CAS across substances and cultures. METHODS: This study analyzed the CAS-scores of a sample of 370 participants in Germany, China and the UK, including 262 opioid-users, 65 smokers and 43 alcohol-users. We performed stability analyses to check the stability (i. e. factorial invariance) of factor solutions. Based on confirmed stability of the general factor (gfactor) solution and the calculations rule obtained in the previous validation of CAS-alcohol (CAS-A), the factor structures of CAS-A, CAS-S and CAS-CS were compared. RESULTS: The gfactor solutions based on calculations rule shows good stability, with the mean stability coefficients of 0.990 and 0.977 for CAS-S and CAS-CS respectively. The gfactor patterns were similar for CAS-A, CAS-S and CAS-CS, as well as across samples (Germany, China and the UK), with most factor-loadings larger than 0.7. Based on these findings, CAS-S and CAS-CS were also associated with established clinical measures of SUD. CONCLUSIONS: Our findings suggest the two-gfactor solution based on a proposed calculation rule has a high stability across substances and cultures. This could be in line with common neurobiological mechanisms underlying habitual substance use. Moreover, comparing CAS with established clinical tools suggests that CAS might assess the automated behavior in substance consumption in a more sophisticated way.

• The two-gfactor solution of the Craving Automated Scale has a good stability.• The Craving Automated Scale can be used across substances.• The Craving Automated Scale is associated with established SUD measures.

Flavor-specific enhancement of electronic cigarette liquid consumption and preference in mice.

BACKGROUND: The use of electronic cigarettes has increased over the past decade. To determine how the abuse liability of electronic cigarette liquids (e-liquids) differs from nicotine alone, and to determine the impact of flavor, we compared nicotine-containing fruit- and tobacco-flavored e-liquids, and their nicotine-free versions, to nicotine alone in mouse models of oral consumption, reward and aversion. METHODS: Adult male C57BL/6 J mice voluntarily consumed oral nicotine, equivalent nicotine concentrations of fruit- and tobacco-flavored e-liquid, and equivalent dilutions of the nicotine-free versions in 2-bottle choice tests. Conditioned place preference and place aversion were assessed with peripherally administered e-liquids or nicotine. Serum nicotine and cotinine levels were measured after subcutaneous injections of e-liquid or nicotine. RESULTS: Mice showed higher consumption and preference for the fruit-flavored e-liquid compared with nicotine alone. This increase was not due to the flavor itself as consumption of the nicotine-free fruit-flavored e-liquid was not elevated until the highest concentration tested. The increased consumption and preference were not observed with the tobacco-flavored e-liquid. The conditioned place preference, place aversion and nicotine pharmacokinetics of the fruit-flavored e-liquid were not significantly different from nicotine alone. CONCLUSIONS: Our data suggest that fruit, but not tobacco flavor, increased the oral consumption of e-liquid compared with nicotine alone. Moreover, this enhancement was not due to increased consumption of the flavor itself, altered rewarding or aversive properties after peripheral administration, or altered pharmacokinetics. This flavor-specific enhancement suggests that some flavors may lead to higher nicotine intake and increased use of e-liquids compared with nicotine alone.

Is sitting time leading to mobility decline in long-term care residents?

BACKGROUND: Little is known about the sitting time in long-term care (LTC) facilities, or if sedentary behaviour affects changes in mobility over time. AIMS: The objectives were to document the sitting time of LTC residents and to examine if sitting time could predict changes in mobility. METHODS: Twenty residents of an LTC facility, representing three mobility statuses (independent, assisted transfer, and dependent transfer) were included. Sitting time was defined using an ActivPAL. Mobility statuses were reviewed 12 months later. RESULTS: Participants spent an average of 21.9 h per day sedentary. At follow-up, five residents experienced a decline in mobility status, but no baseline sitting time variables were associated with the changes in mobility status (P > 0.05). DISCUSSION/CONCLUSION: People living in LTC are highly sedentary. Sitting time differs amongst the mobility statues, but is unable to predict upcoming changes in mobility status.

Taq1a polymorphism (rs1800497) is associated with obesity-related outcomes and dietary intake in a multi-ethnic sample of children.

BACKGROUND: In adults, the Taq1a polymorphism (rs1800497) near the D2 receptor (DRD2) gene is associated with body mass index and binge eating and is more prevalent among non-Hispanic Blacks (NHB) and Hispanic-Americans (HA) relative to non-Hispanic Whites (NHW). We hypothesize Taq1a polymorphism (rs1800497) risk alleles contribute to paediatric racial/ethnic differences in obesity phenotypes. OBJECTIVES: This study aims to characterize the relationship between the Taq1a polymorphism (rs1800497), diet and adiposity in a multi-ethnic cohort of 286 children (98 NHB, 76 HA and 112 NHW), ages 7-12. METHODS: Dual-energy X-ray absorptiometry, computed tomography scans and two 24-h dietary recalls assessed body composition, fat distribution and dietary intakes, respectively. RESULTS: Children with two Taq1a risk alleles (NHB = 50.0%, HA = 43.3%, NHW = 6.7%) reported a 20% increase in total energy intake (P = 0.0034) and per cent of calories from sugar consumed (P = 0.0077) than did children with less than two risk alleles. Children with two Taq1a risk alleles demonstrated significantly higher total body fat (P = 0.0145), body fat percentage (P = 0.0377), intra-abdominal adiposity (P = 0.0459), subcutaneous abdominal adiposity (P = 0.0213) and total abdominal adiposity (P = 0.0209) than did children with one or no Taq1a risk alleles. CONCLUSIONS: Our results suggest that having two Taq1a risk alleles is associated with an increase in reported calorie and sugar consumption and is a potential risk factor for early development of excess adiposity in multi-ethnic children. These results need to be confirmed in a larger sample.

Elevated preoperative heart rate is associated with cardiopulmonary and autonomic impairment in high-risk surgical patients.

BACKGROUND: Elevated preoperative heart rate (HR) is associated with perioperative myocardial injury and death. In apparently healthy individuals, high resting HR is associated with development of cardiac failure. Given that patients with overt cardiac failure have poor perioperative outcomes, we hypothesized that subclinical cardiac failure, identified by cardiopulmonary exercise testing, was associated with elevated preoperative HR > 87 beats min -1 (HR > 87). METHODS: This was a secondary analysis of an observational cohort study of surgical patients aged ≥45 yr. The exposure of interest was HR > 87, recorded at rest before preoperative cardiopulmonary exercise testing. The predefined outcome measures were the following established predictors of mortality in patients with overt cardiac failure in the general population: ventilatory equivalent for carbon dioxide ( V˙E/V˙co2 ) ratio ≥34, heart rate recovery ≤6 and peak oxygen uptake ( V˙o2 ) ≤14 ml kg -1  min -1 . We used logistic regression analysis to test for association between HR > 87 and markers of cardiac failure. We also examined the relationship between HR > 87 and preoperative left ventricular stroke volume in a separate cohort of patients. RESULTS: HR > 87 was present in 399/1250 (32%) patients, of whom 438/1250 (35%) had V˙E/V˙co2 ratio ≥34, 200/1250 (16%) had heart rate recovery ≤6, and 396/1250 (32%) had peak V˙o2 ≤14 ml kg -1  min -1 . HR > 87 was independently associated with peak V˙o2 ≤14 ml kg -1  min -1 {odds ratio (OR) 1.69 [1.12-3.55]; P =0.01} and heart rate recovery ≤6 (OR 2.02 [1.30-3.14]; P <0.01). However, HR > 87 was not associated with V˙E/V˙co2 ratio ≥34 (OR 1.31 [0.92-1.87]; P =0.14). In a separate cohort, HR > 87 (33/181; 18.5%) was associated with impaired preoperative stroke volume (OR 3.21 [1.26-8.20]; P =0.01). CONCLUSIONS: Elevated preoperative heart rate is associated with impaired cardiopulmonary performance consistent with clinically unsuspected, subclinical cardiac failure. CLINICAL TRIAL REGISTRATION: ISRCTN88456378.

Genome-wide association studies of smooth pursuit and antisaccade eye movements in psychotic disorders: findings from the B-SNIP study.

Eye movement deviations, particularly deficits of initial sensorimotor processing and sustained pursuit maintenance, and antisaccade inhibition errors, are established intermediate phenotypes for psychotic disorders. We here studied eye movement measures of 849 participants from the Bipolar-Schizophrenia Network on Intermediate Phenotypes (B-SNIP) study (schizophrenia N=230, schizoaffective disorder N=155, psychotic bipolar disorder N=206 and healthy controls N=258) as quantitative phenotypes in relation to genetic data, while controlling for genetically derived ancestry measures, age and sex. A mixed-modeling genome-wide association studies approach was used including ~4.4 million genotypes (PsychChip and 1000 Genomes imputation). Across participants, sensorimotor processing at pursuit initiation was significantly associated with a single nucleotide polymorphism in IPO8 (12p11.21, P=8 × 10-11), whereas suggestive associations with sustained pursuit maintenance were identified with SNPs in SH3GL2 (9p22.2, P=3 × 10-8). In participants of predominantly African ancestry, sensorimotor processing was also significantly associated with SNPs in PCDH12 (5q31.3, P=1.6 × 10-10), and suggestive associations were observed with NRSN1 (6p22.3, P=5.4 × 10-8) and LMO7 (13q22.2, P=7.3x10-8), whereas antisaccade error rate was significantly associated with a non-coding region at chromosome 7 (P=6.5 × 10-9). Exploratory pathway analyses revealed associations with nervous system development and function for 40 top genes with sensorimotor processing and pursuit maintenance (P=4.9 × 10-2-9.8 × 10-4). Our findings suggest novel patterns of genetic variation relevant for brain systems subserving eye movement control known to be impaired in psychotic disorders. They include genes involved in nuclear trafficking and gene silencing (IPO8), fast axonal guidance and synaptic specificity (PCDH12), transduction of nerve signals (NRSN1), retinal degeneration (LMO7), synaptic glutamate release (SH3GL2), and broader nervous system development and function.

Quantitative Contribution of rs75017182 to Dihydropyrimidine Dehydrogenase mRNA Splicing and Enzyme Activity.

Dihydropyrimidine dehydrogenase (DPD; DPYD gene) variants have emerged as reliable predictors of adverse toxicity to the chemotherapy agent 5-fluorouracil (5-FU). The intronic DPYD variant rs75017182 has been recently suggested to promote alternative splicing of DPYD. However, both the extent of alternative splicing and the true contribution of rs75017182 to DPD function remain unclear. In the present study we quantified alternative splicing and DPD enzyme activity in rs75017182 carriers utilizing healthy volunteer specimens from the Mayo Clinic Biobank. Although the alternatively spliced transcript was uniquely detected in rs75017182 carriers, canonically spliced DPYD levels were only reduced by 30% (P = 2.8 × 10-6 ) relative to controls. Similarly, DPD enzyme function was reduced by 35% (P = 0.025). Carriers of the well-studied toxicity-associated variant rs67376798 displayed similar reductions in DPD activity (31% reduction). The modest effects on splicing and function suggest that rs75017182 may have clinical utility as a predictor of 5-FU toxicity similar to rs67376798.

Endovascular Therapy Research in Lower Limb Peripheral Arterial Disease Published Over a 5-Year Period: Who is Publishing and Where?

INTRODUCTION: Peripheral arterial disease (PAD) is being increasingly managed by endovascular therapies. In this study, we identified the clinical services publishing research as well as the journals of publication over a 5-year period. METHODS: Twenty keywords and phrases related to endovascular intervention were identified, and a literature search was performed through the PubMed database from January 2009 to January 2014. Inclusion criteria were English language, study population more than five patients, and matching the keyword search. Eligible studies were collated into a database and classified by journal of publication, PubMed number, article title, publishing clinical service, type of publication, country of origin, and authors. RESULTS: 825 studies from 114 different journals were identified. 297 papers were excluded. Of the 528 included papers, 204 (39%) were published by Vascular Surgery (VS), 157 (30%) by Interventional Radiology (IR), 101 (19%) by Cardiology, 43 (8%) by Angiology, 6 (1%) by Vascular Medicine, and 17 (3%) from miscellaneous services. 283 (54%) studies originated from Europe, 157 (30%) from North America, 76 (14%) from Asia, 6 from Australia, 3 each from South America and Africa. IR published the most papers on PAD endovascular intervention in Europe with VS second while this trend was reversed in the USA. The 528 papers were published in 98 different journals with retrospective case series (72%), the majority. CONCLUSION: IR continues to play a significant research role in endovascular intervention in PAD, particularly in Europe, and specifically in below the knee intervention, pedal intervention, and drug-eluting technologies.

Radiation safety awareness among medical interns: are EU guidelines being implemented?

BACKGROUND: European recommendations suggest that medical students should be taught radiation safety before entering clinical practice. AIM: The aim of this study was to produce a summative assessment of radiation protection training in medical school in Ireland. MATERIALS AND METHODS: A web-based questionnaire was distributed to the 2014 intern population (n = 683) via network intern-coordinators. The survey encompassed knowledge of radiation dose in X-ray investigations, laws governing the prescribing of radiation and complications of radiation exposure to staff and patients. RESULTS: Response rate was 14.2% (97/683) with all Irish medical schools represented. 64% of interns reported no formal training in radiation safety. 80% correctly identified MRI and 94% US as not posing a radiation risk. 54% identified CT PET as emitting the highest radiation dose to patients. Only 32% correctly identified one CT abdomen/pelvis as equivalent to the dose from 300 to 500 chest X-rays and 22% correctly identified the theoretical lifetime risk of cancer induction from CT abdomen/pelvis as 1 in 2000. While 71% thought it was very important that prescribers should be aware of patient radiation dose and 28% thought it was moderately important, 74% were not aware of any laws governing the prescribing of radiology investigations. CONCLUSION: Currently, there is little formal radiation safety training in Irish medical schools. Knowledge of radiation dose and the laws governing prescribing is limited among qualifying interns. Implementation of a formal radiation safety curriculum in Irish Medical Schools would adhere to EU guidelines and improve prescriber knowledge, patient, and personal radiation safety.

Survey on examining prevalence of paternal anxiety and its risk factors in perinatal period in Hong Kong: a longitudinal study.

BACKGROUND: There is emerging evidence of the significance of paternal mental health problems among the expectant fathers during the antenatal and postnatal period. The present study aims at determining the prevalence of paternal perinatal anxiety and identifying its risk factors among the fathers. METHODS: A total of 622 expectant fathers were recruited in Hong Kong. The expectant fathers were assessed using standardized and validated psychological instruments on three time points including early pregnancy, late pregnancy and 6 week postnatal. Independent samples t-test, one way ANOVA, Pearson's correlation and multiple linear regression were used to examine the effect of hypothesized risk factors. Hierarchical multiple regression and mixed effect model were also conducted with potential confounding factors controlled for. RESULTS: Results showed that a significant proportion of expectant fathers experienced anxiety during the perinatal period. Low self-esteem and poor social support were found to be risk factors of paternal anxiety across pregnancy to postnatal period. Work-family conflict could significantly predict paternal anxiety in the pregnancy period. CONCLUSIONS: The present study points to the need for greater research and clinical attention to paternal anxiety, given that it is a highly prevalent problem and could be detrimental to their partner's well-being and children development. The present findings contributes to the theoretical understanding of the prevalence and risk factors of paternal perinatal anxiety and have implications for the design of effective identification, prevention, and interventions of these clinical problems.

PKCε phosphorylates α4ß2 nicotinic ACh receptors and promotes recovery from desensitization.

BACKGROUND AND PURPOSE: Nicotinic (ACh) receptor recovery from desensitization is modulated by PKC, but the PKC isozymes and the phosphorylation sites involved have not been identified. We investigated whether PKCε phosphorylation of α4ß2 nAChRs regulates receptor recovery from desensitization. EXPERIMENTAL APPROACH: Receptor recovery from desensitization was investigated by electrophysiological characterization of human α4ß2 nAChRs. Phosphorylation of the α4 nAChR subunit was assessed by immunoblotting of mouse synaptosomes. Hypothermia induced by sazetidine-A and nicotine was measured in Prkce(-/-) and wild-type mice. KEY RESULTS: Inhibiting PKCε impaired the magnitude of α4ß2 nAChR recovery from desensitization. We identified five putative PKCε phosphorylation sites in the large intracellular loop of the α4 subunit, and mutating four sites to alanines also impaired recovery from desensitization. α4 nAChR subunit phosphorylation was reduced in synaptosomes from Prkce(-/-) mice. Sazetidine-A-induced hypothermia, which is mediated by α4ß2 nAChR desensitization, was more severe and prolonged in Prkce(-/-) than in wild-type mice. CONCLUSIONS AND IMPLICATIONS: PKCε phosphorylates the α4 nAChR subunit and regulates recovery from receptor desensitization. This study illustrates the importance of phosphorylation in regulating α4ß2 receptor function, and suggests that reducing phosphorylation prolongs receptor desensitization and decreases the number of receptors available for activation.

The Prevalence and Risk Factors of Paternal Depression from the Antenatal to the Postpartum Period and the Relationships between Antenatal and Postpartum Depression among Fathers in Hong Kong.

Introduction. Despite the fact that maternal perinatal mental health problems have been extensively studied and addressed to be a significant health problem, the literature on paternal perinatal mental health problems is relatively scarce. The present study aims at determining the prevalence of paternal perinatal depression and identifying the risk factors and the relationship between antenatal and postpartum depression. Methodology. 622 expectant fathers were recruited from regional maternal clinics. The expectant fathers were assessed using standardized and validated psychological instruments on 3 time points including early pregnancy, late pregnancy, and six weeks postpartum. Results. Results showed that a significant proportion of expectant fathers manifested depressive symptoms during the perinatal period. Paternal antenatal depression could significantly predict higher level of paternal postpartum depression. Psychosocial risk factors were consistently associated with paternal depression in different time points. Conclusions. The present study points to the need for greater research and clinical attention to paternal depression given that it is a highly prevalent problem and could be detrimental to their spouse and children development. The present findings contribute to theoretical basis of the prevalence and risk factors of paternal perinatal depression and have implications of the design of effective identification, prevention, and interventions of these clinical problems.

A DPYD variant (Y186C) in individuals of african ancestry is associated with reduced DPD enzyme activity.

5-Fluorouracil (5-FU) is used to treat many aggressive cancers, such as those of the colon, breast, and head and neck. The responses to 5-FU, with respect to both toxicity and efficacy, vary among racial groups, potentially because of variability in the activity levels of the enzyme dihydropyrimidine dehydrogenase (DPD, encoded by the DPYD gene). In this study, the genetic associations between DPYD variations and circulating mononuclear-cell DPD enzyme activity were evaluated in 94 African-American and 81 European-American volunteers. The DPYD-Y186C variant was unique to individuals of African ancestry, and DPD activity was 46% lower in carriers as compared with noncarriers (279 ± 35 vs. 514 ± 168 pmol 5-FU min(-1) mg(-1); P = 0.00029). In this study, 26% of the African Americans with reduced DPD activity were carriers of Y186C. In the African-American cohort, after excluding Y186C carriers, homozygous carriers of C29R showed 27% higher DPD activity as compared with noncarriers (609 ± 152 and 480 ± 152 pmol 5-FU min(-1) mg(-1), respectively; P = 0.013).

Intestinal failure and home parenteral nutrition: implications for oral health and dental care.

BACKGROUND & AIMS: Background disease processes, medication and therapies in people with intestinal failure receiving home parenteral nutrition may affect their oral health. To inform oral health advice for this group a study of their oral health status was carried out. METHODS: Fifty-two HPN outpatients recruited from specialised nutrition clinics at a national referral centre listed their medical and medication history, perceived oral health and dental treatment experience in a structured interview and underwent an oral health examination. Findings were compared with 2009 UK Adult Dental Health Survey data, using one-sample t tests. RESULTS: Oral health of the HPN cohort was poorer than the UK norm; patients had more decay (p<0.001), fewer teeth (p<0.001) and fewer sound and untreated teeth (p=0.023) despite similar dental attendance. Hyperphagia, sip feeds, oral rehydration fluids and polypharmacy (in 96%) are identifiable risk factors for caries, xerostomia (in 81%) and thus oral infection risk (including oral candidiasis). Patients were experiencing current problems (60%) and psychological discomfort (56%) from poor oral health. The patient pathway does not include oral health information. CONCLUSION: Dental teams should be aware of the management and prevention of HPN related complications with bisphosphonates, anticoagulant therapy, and parenteral antibiotic prophylaxis. HPN patients may benefit from increased awareness of their oral health risk factors.