Cindy L. Ehlers

P300 latency reflects the degree of cognitive decline in dementing illness.

An auditory discrimination paradigm was employed to elicit the P3 component of the event-related brain potential (ERP) from 39 demented patients (mean age = 71 years). Component latency was longer in patients who were diagnosed as having primary degenerative dementia and other cognitive impairment disorders compared to age-matched controls. Neurologist ratings of cognitive impairment were significantly correlated with P3 latency values, although no differences in mean latency were obtained between the various categories of dementia. ERP measurement techniques and the interpretation of P3 latency as in index of dementing illness are discussed.

Addictions Biology: Haplotype-Based Analysis for 130 Candidate Genes on a Single Array

AIMS To develop a panel of markers able to extract full haplotype information for candidate genes in alcoholism, other addictions and disorders of mood and anxiety. METHODS A total of 130 genes were haplotype tagged and genotyped in 7 case/control populations and 51 reference populations using Illumina GoldenGate SNP genotyping technology, determining haplotype coverage. We also constructed and determined the efficacy of a panel of 186 ancestry informative markers. RESULTS An average of 1465 loci were genotyped at an average completion rate of 91.3%, with an average call rate of 98.3% and replication rate of 99.7%. Completion and call rates were lowered by the performance of two datasets, highlighting the importance of the DNA quality in high throughput assays. A comparison of haplotypes captured by the Addictions Array tagging SNPs and commercially available whole-genome arrays from Illumina and Affymetrix shows comparable performance of the tag SNPs to the best whole-genome array in all populations for which data are available. CONCLUSIONS Arrays of haplotype-tagged candidate genes, such as this addictions-focused array, represent a cost-effective approach to generate high-quality SNP genotyping data useful for the haplotype-based analysis of panels of genes such as these 130 genes of interest to alcohol and addictions researchers. The inclusion of the 186 ancestry informative markers allows for the detection and correction for admixture and further enhances the utility of the array.

A decrease in the size of the basal ganglia following prenatal alcohol exposure: A preliminary report

Prenatal alcohol exposure is known to cause damage to the central nervous system. This study sought to further elucidate the structural brain damage that occurs following prenatal alcohol exposure in both children and rats. Two children with histories of maternal alcohol abuse but who did not qualify for a diagnosis of Fetal Alcohol Syndrome (FAS), based on established criteria, underwent magnetic resonance imaging. Reduced volumes were found for the cerebrum and cerebellum. In addition, the proportional volume of the basal ganglia was reduced, although the proportional volumes of cortical and subcortical fluid, cortical gray matter, limbic and nonlimbic cortex, and diencephalic structures were unaffected. These findings are compared with our recent MRI findings in two cases of FAS. In addition, the caudate-putamen and ventricular areas were assessed in rats exposed to alcohol prenatally. Whereas the overall brain section area was not reduced in size, the area of the caudate-putamen was reduced and that of the ventricles was enlarged.

Slow-wave sleep: do young adult men and women age differently?

The differential effects of ageing on polysomnographic and EEG spectral characteristics of sleep were explored in men and women between the ages of 20 and 40. Men and women in their twenties were found to have similar percentages of slow‐wave sleep (SWS) (% Stage 3 and 4) and mean EEG slow wave activity (quantified by spectral analysis). Significant reductions in the percentage of SWS and mean slow wave activity over the night occurred in men during their thirties but not in the women. This suggests that gender difference in SWS may emerge between age 30 and 40 in young adults. Men in this sample were also found to have significant increases in Stage 2 sleep, and decreases in REM sleep time, REM activity, REM density and REM intensity. No significant effects of age were found for women in any visually scored sleep variables. Both men and women had age related reductions in spectral power in the spindle frequencies. Taken together, these findings suggest that the sleep of men and women over age 20–40 may age differently.

Verbal Learning Differences in Epileptic Patients with Left and Right Temporal Lobe Foci

Summary: A verbal learning test patterned after and using the same format as the Rey Auditory‐Verbal Learning Test was administered to the following three groups: (1) patients with left temporal lobe epilepsy (L‐TLE) as defined by EEG criteria (n = 11); (2) patients with right temporal lobe epilepsy (n = 10); and (3) normal controls (n = 11). Performance was highly similar for all three groups during the five immediate recall learning trials. The performance of the L‐TLE group, averaged across three delayed recall trials (free recall, phonemic cued recall, semantic cued recall), was significantly poorer than that of the other two groups. The L‐TLE group showed the worst performance on the phonemic cued recall trial, poor performance on the delayed free recall trial, and relatively intact performance on the semantic cued recall trial. Immediate and delayed free recall and phonemic and semantic cued recall for the distractor list did not discriminate groups. Word frequency, word presentation order, and concrete versus abstract words did not have different effects across groups.

Genomic screen for loci associated with alcohol dependence in Mission Indians

Alcohol dependence is a leading cause of morbidity and mortality in Native Americans, yet biological factors underlying the disorder in this ethnic group remain illusive. This studys aims were to map susceptibility loci for DSM‐III‐R alcohol dependence and two narrower alcohol‐related phenotypes in Mission Indian families. Each participant gave a blood sample and completed an interview using the Semi‐Structured Assessment for the Genetics of Alcoholism (SSAGA) that was used to make alcohol dependence diagnoses and the narrower phenotypes of withdrawal, and drinking severity. Genotypes were determined for a panel 791 microsatellite polymorphisms. Analyses of multipoint variance component LOD scores for the dichotomous DSM‐III‐R phenotype revealed no peak LOD scores that exceeded 2.0 at any chromosome location. Two chromosomes, 4 and 12, had peak LOD scores that exceeded 2 for the alcohol use severity phenotype and three chromosomes 6, 15, 16 were found to have peaks with LOD scores that exceeded 2 for the withdrawal phenotype. Evidence for linkage to chromosomes 4 and 15, and 16 have been reported previously for alcohol related phenotypes whereas no evidence has as yet been reported for chromosomes 6 and 12. Combined linkage and association analysis suggest that alcohol dehydrogenase 1B gene polymorphisms are partially responsible for the linkage result on chromosome 4 in this population. These results corroborate the importance of several chromosomal regions highlighted in prior segregation studies in alcoholism and further identify new regions of the genome that may be unique to either the restricted phenotypes evaluated or this population of Mission Indians.

Corticotropin releasing factor (CRF): studies in alcohol preferring and non-preferring rats*

Electroencephalographic (EEG) responses to corticotropin releasing factor (CRF) as well as CRF concentrations in several brain regions were measured in two lines of rats which have been genetically selected for alcohol preferring (P) or non-preferring (NP) behaviors. Fifteen rats were implanted with chronic electrodes and EEG spectra were evaluated following intracerebroventricular (ICV) administration of CRF (0.15 nmol) or saline. P rats demonstrated a significantly increased EEG response to CRF in the theta frequency range (ANOVA: PREF × DRUG 4–6 Hz,P<0.03; 6–8 Hz,P<0.05) in frontal cortex. A significantly lower concentration of CRF was found in the P rats in hypothalamus (P<0.02), amygdala (P<0.003), prefrontal cortex (P<0.01), and cingulate cortex (P<0.02). The finding that P rats had an increased response to exogenously administered CRF, taken together with decreased CRF concentrations, suggests that CRF receptors may be up-regulated in these animals. Differences in the regulation of CRF neurons may contribute to the expression of behavioral preference for ethanol consumption in these rat lines.

Alcohol Metabolism in Asian-American Men with Genetic Polymorphisms of Aldehyde Dehydrogenase

Epidemiologic studies have found that rates of alcohol use and alcoholism in persons of Asian descent are lower than rates in other ethnic groups. One possible reason is that about half of certain Asians, including Chinese, Japanese, and Korean persons, have a deficiency of the low-Km mitochondrial aldehyde dehydrogenase (ALDH2) isoenzyme, which is responsible for metabolizing acetaldehyde. A deficiency of ALDH2 results from inheritance of the mutant ALDH2*2 allele, a dominant mutation that exerts its effect both by reducing enzyme activity and increasing the turnover of this activity [1, 2]. After ingestion of alcohol, the faces of Asians with one or both alleles of ALDH2*2 become visibly flushed. Asians who are homozygous for ALDH2*1 generally lack visible alcohol-induced flushing or experience only a mild flush response. The dominance of the ALDH2 mutation, however, does not seem to be complete; phenotypic differences are associated with the three ALDH2 genotypes. Asians who are homozygous for ALDH2*2 drink very little alcohol [3], and no studies have found alcoholic persons with this genotype [4-8]. Asians who are heterozygous for ALDH2*2 drink less alcohol and are also less likely to be alcoholic compared with Asians with ALDH2*1 alleles, but they are not fully protected from alcoholism. Approximately 12% of alcoholic Asians have the ALDH2*1/2*2 genotype [5]. In the context of alcoholism or lower alcohol intake, Asian persons who are heterozygous for ALDH2*2 may be more vulnerable to alcohol-associated conditions, including liver disease [6-9], asthma [9], and esophageal cancer [10]. The three ALDH2 genotypes are also associated with variability in response to alcohol [11]. Asians who are homozygous for ALDH2*2 are very sensitive to alcohol and have tachycardia, hypotension, and vomiting after ingesting a moderate amount of alcohol. Asians who are heterozygous for ALDH2*2 are more sensitive to alcohol than Asians with ALDH2*1 alleles, although the response of the former is not necessarily aversive. Among Asians with an ALDH2 deficiency, differences in sensitivity to alcohol may be mediated by differences in alcohol metabolism, slower elimination of alcohol, or accumulation of acetaldehyde in the blood [12]. Some studies [13-17] have measured blood levels of alcohol or acetaldehyde after ingestion of alcohol in Asians who were known to have ALDH2 genotypes, but these studies had an inadequate sample size, did not include a placebo control, or did not control for use of alcohol and cigarettes (which can alter alcohol metabolism). We measured blood levels of alcohol and acetaldehyde after ingestion of alcoholic or placebo beverages in Asian-American men who underwent genotyping at the ALDH2 locus. Particular attention was given to matching the groups for age, height, weight, history of alcohol use, and history of smoking. Methods Asian-American men 21 to 25 years of age were recruited from advertisements in university newspapers for our randomized, double-blind, crossover study. They completed a questionnaire that solicited information on demographic characteristics; patterns of and problems with alcohol and drug use; and family history of alcohol, drug, and psychiatric problems. We excluded persons who completely abstained from alcohol, persons who had consumed more than 60 standard alcoholic drinks per month during the previous 6 months, and persons who reported that either biological parent was not of Chinese, Japanese, or Korean descent. Thirty-five men who did not have a personal or family history of alcohol dependence and who had no evidence of other substance dependence, major psychiatric disorders, or medical disorders gave informed consent to participate in two test sessions. The study was approved by the institutional review board at the Scripps Clinic and Research Foundation. Participants were asked to refrain from using alcohol, cigarettes, and other drugs (including aspirin, nonsteroidal anti-inflammatory agents, and anti-histamines) that might alter alcohol metabolism for 3 days before testing. On test days, each participant arrived at the clinic at 7:30 a.m. after an overnight fast. He then ate a low-fat breakfast (two slices of dry toast and juice), and an indwelling heparin lock was inserted for drawing blood. At the first session, blood was drawn and genotyping at the ALDH2 locus was done by using polymerase chain reaction of DNA and allele-specific oligonucleotide probes [1]. At 9:00 a.m., each participant was given a placebo beverage (3 mL of 95% alcohol in a reservoir on top of noncaffeinated, sugar-free soda) or 0.75 mL of 95% alcohol (0.56 g/kg of body weight) as a 20%-by-volume solution in the same mixer. The alcohol and placebo were ingested over 7 minutes through a placebo alcohol apparatus [18]. Blood was drawn to determine levels of alcohol and acetaldehyde before beverage ingestion and 15, 30, 45, 60, 90, 120, and 150 minutes after beverage ingestion. Blood alcohol concentrations were determined by using a modified alcohol dehydrogenase assay [19]. The rate of alcohol elimination (mg/kg per hour) was calculated from the slope of the pseudolinear decline of the blood alcohol concentration-time curve (usually from the 90-, 120-, and 150- minute samples) by using linear least-squares regression. Blood acetaldehyde levels were determined by using a modified fluorigenic high-performance liquid chromatographic assay [20] that had a detection sensitivity in the picomole range and intra-assay and interassay precisions of 2.4% and 3.7%, respectively. Statistical analyses, done by using SYSTAT software (SYSTAT, Inc., Evanston, Illinois), focused on differences between participants with ALDH2*1/2*1 and those with ALDH2*1/2*2. Demographic information, data on recent alcohol and cigarette use, peak blood alcohol concentration, time to peak blood alcohol concentration, volume of distribution, and rate of alcohol elimination were analyzed by using one-way analysis of variance; ALDH2 genotype was a between-participant variable. Data on blood alcohol concentration and acetaldehyde level were analyzed by using separate 2 8 analysis of variance for alcohol and placebo sessions; ALDH2 genotype was a between-participant variable, and time was a repeated measurement. Significant interactions were then analyzed by using post hoc comparisons with contrast matrices. The Bonferroni correction was used to limit the familywise error rate to 0.05 for comparisons among the placebo session time points and among the alcohol session time points. Results Genotyping for ALDH2 revealed 20 participants who had ALDH2*1/2*1 genotype, 13 who had ALDH2*1/2*2 genotype, and 2 who had ALDH2*2/2*2 genotype. Three participants (1 with ALDH2*1/2*2 genotype and the 2 with ALDH2*2/2*2 genotype) became ill after ingesting alcohol and were excluded from analyses because of missing data. One participant with ALDH2*1/2*2 genotype whose acetaldehyde levels exceeded 4 SDs from the mean (most likely as a result of instrumentation error) was also excluded from data analyses. The Table 1 shows demographic information and patterns of recent alcohol and cigarette use for the remaining 31 men. The ALDH2 genotype groups did not differ significantly for any of these variables; this reflects participant selection. Table 1. Demographic Information and Recent Patterns of Alcohol and Cigarette Use in 20 Asian-American Men with ALDH2*1/2*1 Genotype and 11 Asian-American Men with ALDH2*1/2*2 Genotype* Mean peak blood alcohol concentration SD was 81.3 12.48 mg/dL; the peak occurred 43.1 15.85 minutes after ingestion of alcohol. Mean volume of distribution was 0.718 0.1124 L/kg of body weight, and the mean rate of alcohol elimination was 97.8 33.97 mg/kg per hour. The ALDH2 genotype groups did not differ significantly for any of these variables. Mean blood alcohol concentrations for the alcohol session and mean acetaldehyde levels for the placebo and alcohol sessions, measured over time according to ALDH2 genotype, are shown in the (Figure 1). Figure 1. Mean (SD) blood levels of alcohol and acetaldehyde before and after ingestion of a placebo beverage containing 3 mL of 95% alcohol and an alcoholic beverage (0. n n P Analysis of variance revealed that the main effects for ALDH2 genotype and the interaction between ALDH2 genotype and time were not significant for the data on blood alcohol concentration from the alcohol session. Analysis of variance also revealed that the main effects for ALDH2 genotype and the interaction between ALDH2 genotype and time were significant for the data on acetaldehyde levels from the placebo sessions (ALDH2 genotype, P < 0.005; interaction between ALDH2 genotype and time, P < 0.013) and the alcohol sessions (ALDH2 genotype, P < 0.002; interaction between ALDH2 genotype and time, P < 0.001). Post hoc analyses with Bonferroni corrections revealed significant group differences 30, 45, and 60 minutes after placebo ingestion and 60, 90, 120, and 150 minutes after alcohol ingestion. Discussion We found that Asian-American men who were heterozygous for the ALDH2*2 allele did not differ from carefully matched men who were homozygous for the ALDH2*1 allele in measures of blood alcohol concentration overall or at any time after alcohol ingestion. These findings are consistent with the results of one study [14] but differ from those of other studies [15-17] in which persons with ALDH2*2 alleles had significantly slower rates of alcohol elimination than did persons with ALDH2*1 alleles. These discrepancies may result from group differences on important variables, especially history of alcohol use, that we controlled for. We also found that, despite equivalent blood alcohol concentrations, participants with ALDH2*1/2*2 genotype had significantly higher blood acetaldehyde levels after ingesting the alcohol beverage than did participants with ALDH2*1/2*1 genotype. These findings suggest that blood acetaldehyde levels rather than bloo

Hypothalamic peptide modulation of EEG sleep in depression: A further application of the S-process hypothesis

Several lines of evidence suggest that a constellation of electroencephalogram (EEG) monitored sleep abnormalities exists in the vast majority of depressed patients. Disturbances of sleep during depressive episodes can include sleep maintenance difficulties, a delay in sleep onset, and a reduced amount of slow-wave sleep (Kupfer et al. 1984). Rapid eye movement (REM) sleep alterations during depressive episodes include a shortened first NREM period (which is usually referred to as a decrease in REM latency) and an increase in REM density during the first few hours of sleep, especially in the first REM period. The majority of these sleep abnormalities, whether they represent changes in REM or NREM sleep, are most pronounced during the first 100-120 min following sleep onset. creases in cortisol secretion rates (Linkowski et al. 1985), a flattening of the circadian rhythm in cortisol (Sachar 1976), and elevated cortisol nadir (Jarrett et al. 1983) have been reported and replicated in numerous investigations. The failure to suppress plasma cortisol in the Dexamethasone Suppression Test (DST) in many depressed patients has also supported the notion of a specific HPA abnormality (Carroll 1982).

CRF/NPY interactions: a potential role in sleep dysregulation in depression and anxiety.

Neuropeptide Y (NPY) has neuromodulatory actions on multiple brain functions including endocrine, behavioral, and circadian processes and has been implicated in the pathophysiology of both anxiety and depression. Behavioral studies suggest that NPY is a potent anxiolytic, whereas CRF is anxiogenic, thus it seems that a balance of these two peptides may exert important influences on behavioral state regulation. However, little is known about how the NPY/CRF balance affects general arousal, attention, and/or sleep states. The present study evaluated the effects of CRF alone, and co‐administered with NPY, on spontaneous brain activity as well as on auditory processing using electrophysiological measures. Electroencephalographic (EEG) and event‐related potentials (ERPs) were obtained in rats following intracerebroventricular administration of CRF (0.5 μg) and CRF (0.5 μg)/NPY (5.0 or 15 μg). Auditory processing, as assessed by ERPs, was affected most significantly in the frontal cortex where CRF produced increases in the N1 and P3 components of the ERP, and NPY/CRF co‐administration produced significant decreases. These data are consistent with a role for CRF in hyperarousal, and further suggest that NPY may be capable of reversing such states. Administration of CRF also produced a significant increase in the time to sleep onset and a decrease in the amount of time spent in non‐rapid eye movement (NREM) sleep as quantified by scoring the EEG paper records. Co‐administration of NPY with CRF reversed the effects of CRF on sleep duration and sleep onset in a dose‐dependent fashion. Spectral analysis revealed that CRF produced quantitative changes in the EEG that were similar to what has previously been reported. CRF‐induced increases in fast frequency activity were found to be reversed by co‐administration of NPY. Taken together these data suggest that “dysregulation” of sleep and arousal states in depression and anxiety may be consistent with an upset of the balance between hypothalamic neuropeptide systems. Depression and Anxiety 6:1–9, 1997.