Establishment of Cytotoxic and Non-cytotoxic NK-92 Cell Clones - Evaluation of Their Function as Target Cells.

BACKGROUND/AIM: Previous studies have demonstrated that NK cells present in PBMCs might explain why clinical trials conducted with NK-92 as well as CAR modified NK-92 cells have to a large extent failed. Two NK-92 clones with different NK target cell properties have been established and are described here. MATERIALS AND METHODS: Two NK-92 cell clones, NK-92 clone 1 and clone 2, were established using the limiting dilution technique. A time-resolved fluorometric assay (TDA-labeled NK-92 clone 1, 2 or K562 as target cells) was used for measuring their sensitivities to NK cell-mediated cytolysis and their NKG2D expression was identified with immunoblotting. RESULTS: A striking difference between the NK-92 clones in their cytotoxic capacity against K562 cells was observed. A clear correlation was noticed between these NK-92 clones when used as target cells and their ability to kill K562 cells. A 50:1 effector:target ratio (PBMCs:NK-92 clone 1) gave 6.50±5.44% lysis whereas the corresponding value was 39.9±10.0% with NK-92 clone 2 as target cells. Interestingly, incubating PBMCs in medium for longer times slightly potentiated their NK activity also against the NK-92 clone 1 (E:T ratio 50:1), from 2.5±0.88% lysis (24 h pre-incubation time) to 13.7±9.04% (48 h) and 13.8±6.89% (72 h). Immunoblotting with anti-NKG2D antibodies stained an approximately 34 kDa protein band in lysates prepared from NK-92 clone 1 cells, which corresponds to the NKG2D antigen. A very faint band of the same size was observed in lysates prepared from NK-92 clone 2 cells. CONCLUSION: The NK-92 clones 1 and 2, established and described here, might turn out to be very useful for finding possible solutions for using NK-92 and CAR NK-92 cells in future treatments of human malignant diseases.

Human IL-15 Inhibits NK Cells Specific for Human NK-92 Cells.

BACKGROUND/AIM: Recent studies have indicated that natural killer (NK) cells present in peripheral blood mononuclear cells (PBMCs) might be responsible for the somewhat poor outcome of clinical trials conducted with the NK cell line NK-92, as well as chimeric antigen receptor-modified NK-92 cells against leukemias and lymphomas. These NK cells and how their cytotoxic profiles can be altered by some common gamma chain receptor-dependent cytokines or by removal of CD4+ cells have been addressed herein. MATERIALS AND METHODS: A time-resolved fluorometric assay using 2.2':6'.2"-terpyridine-6.6"-dicarboxylic acid-labeled NK-92 or K562 as target cells was used for measuring the cytotoxic activity of cytokine-treated PBMCs and purified NK cells. RESULTS: Pre-incubation with 25 ng/ml interleukin 12 (IL-12), IL-15 or IL-21 for 72 h increased NK cell activity against K562 cells by more than 90% (1:25 target:effector ratio), whereas the corresponding NK cell activity against NK-92 cells was reduced by 15.9±0.1% by IL-12 and 50.6±2.9% by IL-15 compared to cells treated with medium alone. IL-7, on the other hand, increased NK activity against K562 to a much smaller extent (10.4±0.4%) and inhibited NK-92 cell lysis by 15.2±0.3%. Interestingly, similar amounts of IL-2 potentiated NK cell activity against both K562 and NK-92 cells by 50.9±0.5% and 14.3±0.9%, respectively. Purification of NK cells with magnetic beads demonstrated that NK cells indeed were responsible for the observed cytotoxic activity against both NK-92 cells (58.5±9.10%, 1:100 target:effector ratio) and K562 cells (81.6±9.57%, 1:100 target:effector ratio). Elimination of CD4+ cells from PBMCs did not alter the NK activity profile. CONCLUSION: This study highlights a problem that might arise with immune-based NK-92 and chimeric antigen receptor-transduced NK-92 cell therapies and pinpoints the need for evaluating new NK-like cell lines.

Human NK-92 Cells Function as Target Cells for Human NK Cells - Implications for CAR NK-92 Therapies.

BACKGROUND/AIM: Recent studies indicate that chimeric antigen receptor (CAR)-T-cells seem to be superior to CAR modified NK-92 cells. One, at least partial, explanation to this discrepancy has been addressed herein, by having NK-92 cells as target cells in cytotoxicity reactions using peripheral blood mononuclear cells. MATERIALS AND METHODS: A time-resolved fluorometric assay (TDA-labeled NK-92 or K562 as target cells) was used for measuring the cytotoxic activity of blood mononuclear cells (PBMC). RESULTS: The cytotoxic capacity of the NK-92 cells was initially demonstrated by their ability to efficiently kill K562 cells. Interestingly, having PBMC as effector cells rendered the very same NK-92 cells sensitive to NK-cell mediated cytolysis. A 1:100 target:effector ratio gave 34.1% lysis compared to 72.2% lysis for K562 cells. Incubating PBMC for longer times (24 up to 48 h) potentiated their NK-activity against NK-92 cells even more, reaching a level close to that obtained with K562 cells. CONCLUSION: This study pinpoints a severe problem that has to be considered in future immune-based cancer therapies with NK-92 as well as CAR-transduced NK-92 cells.

The AVI-R2: an inventory for a differentiated diagnosis of alcohol problems.

Often in clinical practice, a diagnosis of alcohol dependence or abuse does not provide enough information to establish an adequate treatment plan. Hence, multidimensional instruments have been constructed better to describe the extent and character of a patient's alcohol problem. The purpose of the current article is to present the AVI-R2, a self-report test that provides a standardized and differentiated description of an ongoing drinking problem. The instrument includes 81 items categorized into 16 primary scales that were further summarized into four secondary scales: alcohol dependence, psychological benefits, relational complications and multiple dependence. The article provides reference values and psychometric properties of the Swedish version of the test. Data were based on 270 alcoholic patients recruited from the Centre for Dependency Disorders in Stockholm. Except for one primary scale (social complications due to drinking), the psychometric properties of the test were satisfactory. Recommendations on when and how the test can be used were also provided.

Psychometric properties and norm data of the Swedish version of the NEO-PI-R.

BACKGROUND: Internationally, the NEO Personality Inventory-Revised (NEO-PI-R) is a well established questionnaire for assessment of personality in accordance with the Five Factor Model. The instrument has been translated into many languages including Swedish. AIM: The aim of this study was to make a psychometric evaluation of the Swedish version of NEO-PI-R based on a sample from the general population. METHODS: Postal questionnaires were sent to a random sample of 1250 persons (n = 766 responders). RESULTS AND CONCLUSIONS: The test showed satisfactory internal consistency in the broad factors as well as the facets. A factor analysis indicated that the factors were similar but not identical to those obtained in American studies. In sum, The Swedish version of the NEO-PI-R shows satisfactory psychometric properties and the instrument will continue to be a valuable tool in psychological research and in clinical practice.

Alcohol habits in Sweden during 1997-2009 with particular focus on 2005 and 2009, assessed with the AUDIT: a repeated cross-sectional study.

AIM: This study aimed to survey the changes in alcohol habits during a period with the European Union legal practices. METHODS: Alcohol habits in Sweden were surveyed in the general Swedish population at four timepoints, in 1997, 2001, 2005 and 2009, using the 10-item AUDIT questionnaire. DESIGN: Four separate randomly drawn cross-sectional samples of 1,250 individuals were surveyed at each timepoint. RESULTS: An average of 70% of the sampled individuals responded to the AUDIT questionnaire. Men had higher total AUDIT scores than women in 2005 and 2009, but scores increased among women 61-71 years old and decreased among men 61-71 years old. Younger men and women 17-27 years old decreased their AUDIT-C consumption scores by almost 20% between 2005 and 2009. An analysis of problem drinkers (+8 for men/+6 for women) indicated that a larger proportion of elderly women drank moderately in 2009 compared with 2005, but fewer elderly women drank hazardously. CONCLUSIONS: Earlier increases in drinking levels between 1997 and 2001 may reflect a latent high demand that was restricted by low availability. When availability due to European Union harmonization increased, alcohol consumption followed suit. After a period of adaptation, alcohol consumption appears to have stabilized.

Personality traits and mental health of severe drunk drivers in Sweden.

The present study was intended to investigate personality characteristics and mental health of severe driving under influence (DUI) offenders in a Swedish cohort. More specifically the aim was to investigate the personality traits as assessed by The NEO personality inventory (NEO-PI-R) and aspects of mental health as assessed by the symptom checklist (SCL-90) as compared to the general population. The subjects were 162 severe DUI offenders (with the BAC >0.099%) with an age range of 18-88 years, 143 males and 19 females. It was found that the openness to experience and conscientiousness scales of NEO-PI-R differentiated Swedish DUI offenders from Swedish norm population. The differences between the DUI group and the general population on the on SCL-90 scales were all significant except on the Hostility scale. Two main subtypes of DUI offenders identified were roughly comparable to types I and II alcoholics, as in Cloninger's typology. Among all the scales used (personality traits, psychiatric comorbidity and alcohol use), the only factor that was predictive for future relapses to drunk driving was the factor of depression.

Psychosocial characteristics of drunk drivers assessed by the Addiction Severity Index, prediction of relapse.

AIMS: To investigate psychosocial characteristics and problems of a representative sample of Swedish drunk drivers with special consideration of the gender of the driver; analyze criminal records of the drivers before and after enrollment in the study; identify psychosocial predictors of relapse to drunk driving. METHODS: Psychosocial characteristics were assessed by the Addiction Severity Index (ASI). Alcohol problems were additionally assessed by the Alcohol Use Disorders Identification Test (AUDIT). Criminal records were collected for the five year period before the enrollment, and for the subsequent two years, from Sweden's official crime statistics. RESULTS: About half of the investigated drivers had other psychosocial problems besides the drink driving offence. Female drivers had more alcohol, drug, psychiatric and relational problems, including with parents when growing up, than male drivers, but less previous and subsequent criminality. Heavily drunk drivers (blood alcohol concentration > or = 0.1%) had more problems with alcohol, legal status and employment and support than the other drunk drivers. Problems with legal status, family and social relations and alcohol use increased the risk of relapse in drunk driving, while medical problems seemed to be a protective factor. Different ASI risk factors were identified for relapse in either traffic offences or other crimes. CONCLUSIONS: As well as the drunk driving offence, drunk drivers often have other psychosocial problems, female drivers in particular. Already the blood alcohol concentration per se gives some indication of the psychosocial problem profile of a drunk driver and the ASI profile has some prognostic value for relapse in drunk driving.

Criminal and alcohol problems among Swedish drunk drivers--predictors of DUI relapse.

The prevalence and types of crime offences, as well as predictors of relapse, among drivers suspected of driving under influence (DUI) were investigated. A total of 1830 Swedish DUI drivers responded to the Alcohol Use Disorders Identification Test--AUDIT. Information about previous DUI offences, other traffic offences along with other types of criminal offences was taken from a crime register. A total criminality (including all traffic offences) of 64% in the period of five years before investigation was analyzed. 40% of the sample had other criminality besides traffic violations during that period. 14.3% of the drivers relapsed to DUI in the two-year period after the investigation. In terms of DUI relapse, the following factors were the main predictors: previous traffic violations, previous DUI offences, previous other criminality (frauds or other acts of dishonesty) and detection hours between 12.00 and 19.00. Detection in general traffic controls and high BAC (blood alcohol concentration) when detected were the strongest factors with negative correlation to DUI re-offence.

Alcohol problems among Swedish drunk drivers: differences related to mode of detection and geographical region.

OBJECTIVE: To investigate the prevalence of alcohol problems in a representative sample of Swedish drivers suspected of drunk driving in comparison with control drivers and the general Swedish population in relation to mode and time of detection. Is the time of day or night or the detection mode important for the prevalence of alcohol problems and which are the best predictors for identifying alcohol problems among DUI offenders? METHODS: Two thousand and one hundred drivers (169 females) suspected of DUI offence during 1997-2001 who agreed to respond to the AUDIT questionnaire (Alcohol Use Disorders Identification Test) and 785 control drivers (266 females) not suspected of DUI recruited at general traffic controls were investigated. RESULTS: Both mode and time of detection were found to be important. The greatest impact on the prevalence of alcohol problems emanated from the predictors in the following order: high BAC; unlicensed driving; detection hours between 12.00 and 18.00, and age under 26 years. Age over 55 years and detection in general traffic controls were the two strongest factors negatively correlated to alcohol problems prevalence. The differences between regions with regard to alcohol problems incidence could only partly be explained by police routines and resources. CONCLUSIONS: Mode and time of detection affect the prevalence of DUI offenders with alcohol problems, and to a lesser degree also BAC level. Both the detection mode and the time of detection are significant for the proportion of identified DUIs with alcohol problems. Because the majority of committed DUI offenses are never identified, it is important to optimize the detection strategies of the police with the purpose of minimizing public damage and expenses.

The self-report Drug Use Disorders Identification Test: Extended (DUDIT-E): reliability, validity, and motivational index.

Among clients who have been screened already for drug-related problems, the Drug Use Disorders Identification Test--Extended (DUDIT-E) maps the frequency of illicit drug use (D), the positive (P) and negative (N) aspects of drug use, and treatment readiness (T). D scores correlated with Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition diagnoses among 154 heavy drug users from criminal justice and drug detoxification settings, as well as with urine test results in drug detoxification units. One-week test/retest intraclass correlation coefficients among 92 male prison inmates were .90, .78, .75, and .84 for D, P, N, and T scores, respectively. Cronbach's alpha were .88-.95 for P score, .88-.93 for N score, and .72-.81 for T score. Principal components analysis supported construct validity for P, N, and T scores. T scores were higher in prison treatment units than in motivational and regular units without treatment emphasis. Motivational index scores differentiated between three categories of heavy drug users; they did not differentiate between prisons and unit types, but this corresponded to unclear structural differentiation between units.

Alcohol habits in Sweden during 1997-2005 measured with the AUDIT.

This study aimed to describe changes in Swedish alcohol habits during a period of "harmonization" with European Union alcohol policy. Three random samples collected during the years 1997 (n=997), 2001 (n=893) and 2005 (n=914) were compared. Alcohol habits and hazardous consumption was measured with the AUDIT (Alcohol Use Disorders Identification Test) distributed to the respondents by post. Results were somewhat diverse, but significant changes in alcohol habits occurred among two subgroups of the population: for women and the age group between 28 and 60 years, AUDIT scores peaked in 2001. The results are discussed in relation to the changes made in Swedish alcohol policy during the investigation period.

Evaluation of the Drug Use Disorders Identification Test (DUDIT) in criminal justice and detoxification settings and in a Swedish population sample.

Psychometric properties of the 11-item self-report Drug Use Disorders Identification Test (DUDIT) were evaluated in a sample of heavy drug users from prison, probation, and inpatient detoxification settings, and in a general Swedish population sample. In the drug user sample, the DUDIT predicted drug dependence with a sensitivity of 90% for both DSM-4 and ICD-10 and a respective specificity of 78 and 88%. Reliability according to Cronbach's alpha coefficient was 0.80. In the population sample, 3.1% scored positive on the DUDIT; T-score values are suggested. The DUDIT screens effectively for drug-related problems in clinically selected groups and may prove useful in the context of public health surveys.

Transcription factor AP-2beta genotype associated with anxiety-related personality traits in women. A replication study.

Attempts to link transmitter system genes to certain aspects of personality have been performed. Several monoamine-related gene variants have been investigated. We previously reported an association between a transcription factor activating protein-2beta (AP-2beta) variant and anxiety-related personality traits as estimated by Karolinska Scales of Personality (KSP). To confirm this reported association, we have, in the present study, analysed an enlarged group of healthy volunteers (n = 370) with regard to AP-2beta genotype and personality traits. For estimation of personality traits, individuals completed 5 different personality questionnaires, i.e. Swedish Universities Scales of Personality (SSP), Health-Relevant 5- Factor Personality Inventory (HP5i), Temperament and Character Inventory, the Revised NEO Personality Inventory and KSP. In contrast to men, women having two long AP-2beta alleles displayed lower scores for muscular tension (KSP; F = 10.65, p = 0.0013), somatic trait anxiety (SSP; F = 7.18, p = 0.0081), trait irritability (SSP; F = 4.51, p = 0.032), mistrust (SSP; F = 4.01, p = 0.0468) and negative affectivity (HP5i; F = 10.20, p = 0.0017) than women with at least one short allele. The data presented in this study, together with our previously published data, suggest that AP-2beta intron 2 genotype is associated with low levels of anxiety-related personality traits in women. Hence, these data further suggest the human AP-2beta gene as a novel candidate gene in personality.

Fetus at risk: prevalence of alcohol consumption during pregnancy estimated with a simple screening method in Swedish antenatal clinics.

AIMS: Maternal alcohol consumption is a major health hazard for the fetus. Sweden has an extensive system of public antenatal care clinics, whose mission is to detect and prevent this type of health hazards. However, very few cases of alcohol consumption during pregnancy are detected. The aim of this study was to examine the prevalence of hazardous or harmful alcohol consumption during pregnancy in a consecutive series of Swedish pregnant females. DESIGN, SETTING, PARTICIPANTS AND MEASUREMENTS: The Alcohol Use Disorders Identification Test (AUDIT) was used to collect anonymous data from consecutive pregnant subjects admitted during 1 year to an antenatal clinic in Stockholm, and signing up for parental education offered routinely (n = 1327). Data were obtained from 1101 subjects, typically in pregnancy week 30. A complete AUDIT form was filled out referring to alcohol use during the year prior to pregnancy. A separate form with the consumption items from AUDIT was filled out to report behaviour during pregnancy. FINDINGS: For the year preceding pregnancy, 17% of subjects reported AUDIT scores of 6 or higher, indicating hazardous or harmful alcohol use in women. Few individuals reported scores of 13 or higher (indicating abuse or dependence), but almost half the subjects (46%) reported binge drinking (six standard drinks on a single occasion) once/month or more often, and 6% reported binge drinking on every occasion of alcohol consumption. One-third of the subjects (30%) continued regular alcohol use during pregnancy, and 6% reported consumption two to four times/month. In a logistic regression model, AUDIT scores for the year prior to pregnancy and subject age, but not education level were significant predictors of continued alcohol use during pregnancy. CONCLUSIONS: Alcohol use during pregnancy is more extensive than has been presumed in Sweden. Simple, clinically useful screening methodology detects hazardous consumption during pregnancy in a manner which regular antenatal care does not. If this methodology can be shown to have similar sensitivity when administered under non-anonymous conditions, it should be made part of routine antenatal care.

NURR1 promoter polymorphisms: Parkinson's disease, schizophrenia, and personality traits.

We have previously identified mutations in exon three in NURR1 (NR4A2) in two patients with schizophrenia (SZ) and one patient with bipolar disease with psychotic symptoms. In the present study we analyzed the promoter region of NURR1 and identified five polymorphic sites: three were found to be in strong linkage disequilibrium with a previously identified polymorphic site in the sixth intron. One polymorphism of this haplotype and the two other independent polymorphisms were investigated for their possible association with SZ and Parkinson's disease (PD) by comparing their frequencies in a Swedish material consisting of 134 subjects with SZ and 207 matched controls and 108 subjects with PD and 125 matched controls. Exon 1 was also investigated in our Parkinson and control material but no variances were found. The distributions of the two most informative polymorphisms in the promoter were investigated in an American material as well consisting of 141 subjects with SZ and 139 matched controls. Furthermore, the identified markers were screened for association with putative endophenotypes of SZ in the Swedish material. The distribution of sequence variants among the Swedish controls matched for SZ was investigated with regard to personality. No significant genotype or allelic association of the three sequence variants with SZ or PD was found. Several comparisons regarding endophenotypes or personality indicated association at the 5% confidence level, although correction for multiple testing rendered none of these findings significant. We conclude that the identified polymorphic sites in the human NURR1 are unlikely to be involved in conferring susceptibility for SZ or PD in our patient material.

Hospitalization and mortality succeeding drunk driving and risky driving.

AIMS: The association between drunk driving (DD) and/or risky driving (RD) offences and subsequent hospitalization and mortality was studied during a 25-year period. METHODS: Information about drinking habits and psychosocial factors for the 8122 conscripts from Stockholm County in 1969-1970 was linked to register data on hospitalization, mortality, DD and RD. RESULTS: Analyses comparing background characteristics of DD and RD groups showed that the prevalence of problem behaviour and drug use was highest in the RD group. The relative risk (RR) for hospitalization after DD and/or RD was significantly elevated in multivariate logistic regression analysis for all the studied diagnostic categories (alcohol diagnoses, narcotic diagnoses, suicide attempts, psychoses, E-codes, all diagnoses), and was especially high for alcohol (RR = 7.2) and diagnosis of drug misuse (RR = 9.2). The RR of all hospitalization was 1.5 for the DD group, 1.8 for the RD group, and 1.9 for those who had been sentenced both for drunk driving and risky driving (DRD), all of which were significantly increased. The RR of death was significantly elevated in all three groups. CONCLUSIONS: The results show a significantly increased risk of hospitalization and mortality both in the DD and the RD group. From a public health and traffic safety perspective, this implies a need for developing and implementing better prevention strategies.

[Swedish women have developed more risky and more harmful alcohol drinking habits. A survey of alcohol drinking changes among Swedes between 1997-2001]. / Svenska kvinnor har fått mer riskfyllda och skadligare alkoholvanor. Undersökning av förändringar i svenskarnas alkoholvanor åren 1997-2001.

A random sample of 1,250 persons from the general Swedish population responded to the Alcohol Use Disorders Identification Test, AUDIT. The results (72.5% response rate) were compared to the results from an identical survey made 1997 (79.8% response rate). The factor structure and the internal reliability showed good correspondence with those of 1997, indicating a stable and satisfactory psychometric quality of the Swedish AUDIT version. The female prevalence of hazardous or harmful alcohol use had increased from 11 to 15 percent between 1997 and 2001. Thus, women, particularly 28-38 years old, reported drinking more often and in greater amounts. Furthermore, the prevalence of female teetotallers had decreased from 18 to 13 percent. The corresponding trends among men were not significant but might have been underestimated due to a greater dropout, particularly among young men in the year of 2001.

Two NOTCH4 polymorphisms and their relation to schizophrenia susceptibility and different personality traits.

BACKGROUND: Recently, linkage disequilibrium mapping of the major histocompatibility complex region on the short arm of human chromosome 6 suggested that the NOTCH4 locus is highly associated with schizophrenia. OBJECTIVES AND METHODS: We analysed two polymorphisms in this gene in Swedish schizophrenic patients ( =74) and control subjects ( =135). The NOTCH4 variants were also analysed in schizophrenic patients with regard to subdiagnosis, age at first hospitalization, abuse/dependence of alcohol, solvents, or drugs, previous suicide attempts, extrapyramidal symptoms, treatment with anticholinergic drugs, and response to anti-psychotic drug treatment. Control subjects were scrutinized with regard to personality, another partially heritable trait suggested being of importance in schizophrenia. In addition, two intermediate endophenotypes suggested being of importance in schizophrenia, dopamine D(2) receptor density in striatum and monoamine metabolites in cerebrospinal fluid, respectively, were investigated with regard to the two NOTCH4 variants. RESULTS: There was no significant association between the patients and the controls for the two investigated polymorphisms neither for the parameters analysed in the schizophrenia material. The NOTCH4 SNP2 variant, an A-->G substitution, was associated with the Karolinska Scales of Personality Irritability scale. The NOTCH4 (CTG)(n) variant was associated with the revised NEO personality inventory Extraversion and Activity (E4) scales. However, after correction for multiple testing, no difference remained significant. The results for the endophenotypes and the polymorphisms were non-significant. CONCLUSIONS: The present study does not support that the investigated NOTCH4 variants have a major influence on susceptibility to schizophrenia or related neurobiological traits.

MR volumetry during acute alcohol withdrawal and abstinence: a descriptive study.

AIMS: The brain volume of chronic drinkers is known to partially recover with abstinence from alcohol. To investigate the relative contribution of grey and white brain matter to this process, magnetic resonance imaging and brain tissue segmentation was used to study brain tissue in acute alcohol withdrawal and abstinence in seven alcohol-dependent men. METHODS: The patients were studied on three occasions; within 48 h after the last drink and approximately one month and two and a half months later. Total brain tissue class volumes [grey matter (GM) and white matter (WM) and cerebrospinal fluid (CSF)] were measured. Eleven healthy volunteers were scanned twice to serve as a control group. The alcohol-dependent patients were investigated with regard to drinking variables, neuropsychological performance and blood biochemistry. RESULTS: In the alcohol-dependent patients, intracranial volume and total GM volume did not change between scan occasions, except in a single patient who demonstrated a GM increase of 4.8% (4.2% relative volume) between scans 2 and 3. For all patients, the increase in total WM volume ranged between 1.9 and 22.4% (absolute volumes) and 2.1 and 21.2% (relative volumes). Between scans 2 and 3, the increase in total WM volume ranged between 0.3 and 13.2% (absolute volumes), and between 1.5 and 14.0% (relative volumes). One patient resumed drinking and was investigated a second time during acute withdrawal. In this patient, the measured decrease of 8.1 and 8.5% of absolute and relative WM volumes corresponded to the size of the volume increase between scans 1 and 2. CSF, GM and WM volumes in the healthy subjects were constant over time. CONCLUSIONS: The results demonstrate that changes in brain volume during short-term abstinence in chronic alcohol-dependent patients are confined to the WM. The time limit of WM volume restitution is variable and continues longer than 3 weeks after withdrawal.