A bioinformatic assay for pluripotency in human cells.

Pluripotent stem cells (PSCs) are defined by their potential to generate all cell types of an organism. The standard assay for pluripotency of mouse PSCs is cell transmission through the germline, but for human PSCs researchers depend on indirect methods such as differentiation into teratomas in immunodeficient mice. Here we report PluriTest, a robust open-access bioinformatic assay of pluripotency in human cells based on their gene expression profiles.

Declarative memory performance is associated with the number of sleep spindles in elderly women.

OBJECTIVE: Recent evidence suggests that the sleep-dependent consolidation of declarative memory relies on the nonrapid eye movement rather than the rapid eye movement phase of sleep. In addition, it is known that aging is accompanied by changes in sleep and memory processes. Hence, the purpose of this study was to investigate the overnight consolidation of declarative memory in healthy elderly women. SETTING: Sleep laboratory of University. PARTICIPANTS: Nineteen healthy elderly women (age range: 61-74 years). MEASUREMENTS: We used laboratory-based measures of sleep. To test declarative memory, the Rey-Osterrieth Complex Figure Test was performed. RESULTS: Declarative memory performance in elderly women was associated with Stage 2 sleep spindle density. Women characterized by high memory performance exhibited significantly higher numbers of sleep spindles and higher spindle density compared with women with generally low memory performance. CONCLUSION: The data strongly support theories suggesting a link between sleep spindle activity and declarative memory consolidation.

The cross-sectional GRAS sample: a comprehensive phenotypical data collection of schizophrenic patients.

BACKGROUND: Schizophrenia is the collective term for an exclusively clinically diagnosed, heterogeneous group of mental disorders with still obscure biological roots. Based on the assumption that valuable information about relevant genetic and environmental disease mechanisms can be obtained by association studies on patient cohorts of ≥ 1000 patients, if performed on detailed clinical datasets and quantifiable biological readouts, we generated a new schizophrenia data base, the GRAS (Göttingen Research Association for Schizophrenia) data collection. GRAS is the necessary ground to study genetic causes of the schizophrenic phenotype in a 'phenotype-based genetic association study' (PGAS). This approach is different from and complementary to the genome-wide association studies (GWAS) on schizophrenia. METHODS: For this purpose, 1085 patients were recruited between 2005 and 2010 by an invariable team of traveling investigators in a cross-sectional field study that comprised 23 German psychiatric hospitals. Additionally, chart records and discharge letters of all patients were collected. RESULTS: The corresponding dataset extracted and presented in form of an overview here, comprises biographic information, disease history, medication including side effects, and results of comprehensive cross-sectional psychopathological, neuropsychological, and neurological examinations. With >3000 data points per schizophrenic subject, this data base of living patients, who are also accessible for follow-up studies, provides a wide-ranging and standardized phenotype characterization of as yet unprecedented detail. CONCLUSIONS: The GRAS data base will serve as prerequisite for PGAS, a novel approach to better understanding 'the schizophrenias' through exploring the contribution of genetic variation to the schizophrenic phenotypes.

A flow-cytometric method to investigate glutamate-receptor-sensitivity in whole blood platelets - results from healthy controls and patients with schizophrenia.

Hypofunction of glutamate receptors may contribute to the symptoms of schizophrenia. Human platelets express glutamate receptors and can serve as peripheral surrogate model for neuronal cells. Aim of this study was to establish a fast and sensitive flow-cytometric method to determine the glutamate-dependent kinetics of intracellular calcium ([Ca++]i) mobilization in platelets of schizophrenic patients. Glutamate stimulated [Ca++]i response was measured with a flow-cytometer in anti-CD-41a-labelled whole blood platelets of treated schizophrenic patients (n=18) and controls (n=18). In two control experiments the NMDA-receptor antagonist MK-801 and the dopamine antagonist amisulpride, respectively, were added to probes from healthy subjects. Stimulation with glutamate led dose-dependently to a mobilization of [Ca++]i in both healthy controls and patients. This effect was significantly reduced in patients. In vitro NMDA-antagonism inhibited the glutamate response, whereas dopamine-antagonism had no effect. Our flow-cytometric method allows to measure glutamate-receptor mediated [Ca++]i response in whole blood platelets, without requiring platelet rich preparations. The reduced glutamate-response in the patients was not explained by a direct inhibitory treatment effect. However, further studies with drug naive patients will be necessary to find out whether or not the observed hypoglutamergic function of platelets is endogenous to the disorder.

Changes in CREB phosphorylation and BDNF plasma levels during psychotherapy of depression.

BACKGROUND: The cyclic adenosine monophosphate response element-binding proteins (CREB) and their interaction with brain-derived neurotrophic factor (BDNF) are essential elements in signal transduction pathways important for cellular resilience and neuroplasticity. They play a decisive role in the concept of altered neuroplasticity in major depression. We have previously demonstrated that the increase in phosphorylated CREB (pCREB) in T lymphocytes is significantly associated with clinical improvement in patients treated with antidepressants. In the present study, we focused on patients treated only with psychotherapy to exclude direct pharmacological actions. In addition to pCREB, we also measured the BDNF plasma levels. METHODS: pCREB in T lymphocytes was determined by Western blot; the BDNF plasma levels with solid-phase ELISA. Psychopathology was evaluated with the Hamilton Rating Scale for Depression (HAMD). Thirty patients meeting DSM-IV criteria for major depressive episodes (MDE) were recruited into this 6-week study. They received interpersonal psychotherapy (IPT) twice weekly. RESULTS: After 6 weeks of IPT, 17 patients responded (reduction of > or =50% of baseline HAMD); after 1 week of treatment pCREB increased significantly compared to the nonresponder group. Measurement of the BDNF plasma levels revealed no differences between the responder and nonresponder groups. Furthermore, the correlations between BDNF plasma levels and pCREB were not significant. CONCLUSIONS: The early increase in pCREB is related to treatment response and does not depend on pharmacological interventions or BDNF plasma levels. For the first time, cellular biological markers could be associated with response to psychotherapy.

Visual declarative memory is associated with non-rapid eye movement sleep and sleep cycles in patients with chronic non-restorative sleep.

OBJECTIVE: Sleep contributes to processes of memory, but many questions still remain open. The aim of this study was to test the role of different aspects of sleep for memory performance in a group of patients with chronic non-restorative sleep. METHODS: Forty-two consecutive patients (mean age 40.3 years; 31 women) with non-restorative sleep were included. All subjects underwent polysomnography for diagnostic reasons and obtained the following diagnoses (International Classification of Sleep Disorders, ICSD): psychophysiological or idiopathic insomnia (N=18), paradoxical insomnia (N=13), mild hypersomnia (N=6), and dysthymic disorder (N=5). Patients with sleep-related breathing disorders or restless legs were not included. Prior to polysomnography on the second night and the next morning, neuropsychological tests were performed. Declarative memory was tested by the Rey-Osterrieth Complex Figure Test and a paired associative word list. Procedural learning was assessed by a mirror-tracing skill. RESULTS: Visual declarative memory performance was significantly associated with total sleep time, sleep efficiency, duration of non-rapid eye movement (NREM) sleep and number of NREM-REM sleep cycles, but not with specific measures of REM sleep or slow wave sleep. CONCLUSIONS: Further indications of a role of sleep, and in particular of NREM sleep and sleep organization, for visual declarative memory were found.

Case management for the treatment of patients with major depression in general practices--rationale, design and conduct of a cluster randomized controlled trial--PRoMPT (PRimary care Monitoring for depressive Patient's Trial) [ISRCTN66386086]--study protocol.

BACKGROUND: Depression is a disorder with high prevalence in primary health care and a significant burden of illness. The delivery of health care for depression, as well as other chronic illnesses, has been criticized for several reasons and new strategies to address the needs of these illnesses have been advocated. Case management is a patient-centered approach which has shown efficacy in the treatment of depression in highly organized Health Maintenance Organization (HMO) settings and which might also be effective in other, less structured settings. METHODS/DESIGN: PRoMPT (PRimary care Monitoring for depressive Patients Trial) is a cluster randomised controlled trial with General Practice (GP) as the unit of randomisation. The aim of the study is to evaluate a GP applied case-management for patients with major depressive disorder. 70 GPs were randomised either to intervention group or to control group with the control group delivering usual care. Each GP will include 10 patients suffering from major depressive disorder according to the DSM-IV criteria. The intervention group will receive treatment based on standardized guidelines and monthly telephone monitoring from a trained practice nurse. The nurse investigates the patient's status concerning the MDD criteria, his adherence to GPs prescriptions, possible side effects of medication, and treatment goal attainment. The control group receives usual care--including recommended guidelines. Main outcome measure is the cumulative score of the section depressive disorders (PHQ-9) from the German version of the Prime MD Patient Health Questionnaire (PHQ-D). Secondary outcome measures are the Beck-Depression-Inventory, self-reported adherence (adapted from Moriskey) and the SF-36. In addition, data are collected about patients' satisfaction (EUROPEP-tool), medication, health care utilization, comorbidity, suicide attempts and days out of work. The study comprises three assessment times: baseline (T0) , follow-up after 6 months (T1) and follow-up after 12 months (T2). DISCUSSION: Depression is now recognized as a disorder with a high prevalence in primary care but with insufficient treatment response. Case management seems to be a promising intervention which has the potential to bridge the gap of the usually time-limited and fragmented provision of care. Case management has been proven to be effective in several studies but its application in the private general medical practice setting remains unclear.

[Signals for the initiation of structured diagnostic procedures for depression in primary health care. A practice-relevant evaluation of international guidelines]. / Signalsituationen für den Beginn einer strukturierten Depressionsdiagnostik in der Allgemeinarztpraxis--Eine praxis-kritische Einschätzung internationaler Leitlinien.

BACKGROUND AND OBJECTIVES: Almost every tenth patient of a general practitioner (GP) suffers from depression. However, only 20-25% of these patients are correctly diagnosed during a GP consultation. How do international guidelines for depression in primary care initiate structured diagnostic procedures for depression? METHODS: We performed a systematic literature search on guidelines for the diagnosis of depression with focus on primary care. The quality of the guidelines was rated according to base of evidence, existence of pilot studies, data on implementation, presentation and specificity for primary care settings, and conflict of interest. We also screened whether and how the guidelines comment on the initiation of structured diagnostic procedures for depression. RESULTS: Of the 22 identified guidelines, only 15 address primary care. Only 3 of these were tested in pilot studies, 3 provided data on implementation, 9 were evidence-based. The best guideline (6 out of 6 criteria met) is available in Dutch and established for The Netherlands only. We ranked the guidelines from NHG, VHA and ICSI as very good in terms of methodological quality. They present 'red flags' that initiate structured diagnostic procedures by 'opportunistic screening'. This is followed by the application of a self-rating instrument and an ICD-10-based diagnostic checklist identifying up to 98% of all patients with depression in a given consultation time of 10 minutes on average. CONCLUSION: Based on these criteria a national diagnostic depression guideline should, from our point of view, explicitly include keys such as "red flags" for the initiation of structured diagnostic procedures.

Differential effects of fluoxetine and imipramine on the phosphorylation of the transcription factor CREB and cell-viability.

It has been shown that antidepressants increase the expression of CREB (cAMP-response-element-binding-protein) and BDNF (brain derived neurotrophic factor) in vivo. Apparently inconsistent to these survival-promoting properties for many years antidepressants are known to induce apoptosis in various cell types in vitro. In the present study we evaluated if the antidepressants imipramine and fluoxetine are capable to influence the translational expression and phosphorylation of CREB (pCREB) in cells known to be apoptosis-inducible by antidepressants. We therefore used jurkat cells and quantified apoptosis via propidiumiodid-staining and FACS-analysis. CREB-expression and -phosphorylation was quantified via western blot. Both antidepressants induced apoptosis within 24 h. Fluoxetin starts to induce significant apoptosis at a concentration of 20 microM, whereas imipramine at 100 microM. At these concentrations both antidepressants also increased the phosphorylation of CREB within 6 h. But even in concentrations to low to induce apoptosis both antidepressants still increased CREB-phosphorylation. Treating cells with lowest concentrations only imipramine revealed an increase of CREB-phosphorylation after long-time treatment over 3 weeks. In all experiments overall CREB-expression remained unchanged. In conclusion our experiments indicate that antidepressants are capable to increase CREB-phosphorylation without induction of apoptosis depending on concentration and duration of treatment. We further assume that antidepressants induce CREB-phosphorylation via signal transduction pathways that are different from those inducing apoptosis.

Changes in CREB-phosphorylation during recovery from major depression.

For decades psychiatrists have been looking for biological state markers measurable by easy blood test in order to follow up and predict early on treatment response in patients with major depression. In the present study we investigated whether or not measuring CREB (cAMP-response-element-binding-protein) phosphorylation in peripheral blood T lymphocytes is a state marker of treatment response. CREB is an ubiquitous key-element of intracellular signal transduction cascades and its transcriptional activity depends on phosphorylation at Ser-133. Several studies in animals demonstrated that the transcriptional activity of CREB is up-regulated by antidepressant treatment. Therefore, it has been hypothesized that antidepressant treatment exerts its therapeutic effect by this mechanism. In the present study, we investigated CREB-phosphorylation in T-lymphocytes of 20 patients before and in the end of week one and two of either psychopharmacological or psychotherapeutic treatment. After two weeks, 15 patients fulfilled the criteria of treatment response (i.e. 30% reduction in HAMD score compared to baseline), whereas five patients did not. In the end of week two, the responders showed a significant increase in CREB-phosphorylation (P = 0.018) compared to the non-responders. This was true for all patients with either treatment regimen. In conclusion, these results indicate for the first time that the increase in CREB-phosphorylation might be a molecular state marker for the response to antidepressant treatment.

Morning headaches in patients with sleep disorders: a systematic polysomnographic study.

OBJECTIVES: Patients with sleep disorders suffer more often from headache after awakening than healthy subjects. However, it still is a matter of controversy whether this applies only to patients with sleep apnea syndrome (SAS) or also to patients with other diagnoses of sleep disorders. METHODS: We asked all patients in our sleep laboratory about the frequency of past headaches and also ascertained the occurrence of morning headaches after awakening in the sleep laboratory. Polysomnographic recordings from nights before morning headache were compared with nights without following headache. Four hundred and thirty-two patients with sleep disorders (age range 18-86 years, 37% women) and 30 healthy subjects (age range 24-55 years, 27% women) participated in this prospective study. RESULTS: The reported frequency of past headaches and the frequency of morning headache in the sleep laboratory were significantly increased in patients with SAS and other sleep disorders compared with healthy subjects. The occurrence of morning headache in the sleep laboratory was associated polysomnographically with a decrease in total sleep time, sleep efficiency and amount of rapid eye movement sleep and with an increase in the wake-time during the preceding night. CONCLUSIONS: We conclude that morning headaches in patients with sleep disorders might be associated with particular disturbances of the preceding night's sleep. We speculate that dysregulation in anatomically identical central regions modulating sleep and nociception might be relevant to morning headache, rather than one particular sleep disorder such as SAS.

Novel object presentation affects sleep-wake behavior in rats.

Sleep is suggested to be crucial for the processing and storage of new information. Several learning tasks have been shown to increase the amount of rapid eye movement sleep (REMS) with its typical theta activity (6-8 Hz) relative to total sleep time. Vice versa, REMS deprivation is able to affect memory consolidation following some, but not all learning tasks. Furthermore, recent studies have shown an increase of spindle activity (12-15 Hz) within the electroencephalogram (EEG) of nonREMS as well. The enhancement of both spindle and theta activity is suggested to serve as background activity for the synchronization of those neuronal pathways that were involved in the registration and, later on, participate in the long-term storage of new information in defined brain regions. In the present study, the presentation of a novel object to rats enhanced the amount of preREMS, an intermediate sleep stage with high spindle activity, within the first 2 h of the subsequent sleeping phase. Four hours later, the amount of REMS was increased as well. However, there were no changes in the EEG power spectra of nonREMS, preREMS and REMS. We therefore hypothesize that the increase of preREMS and REMS amounts and the related spindle and theta activity stand for the processing and storage of new information about the presented novel objects.

Perinatal MK-801 treatment affects age-related changes in locomotor activity from childhood to later adulthood in rats.

Neuronal degeneration underlying the pathology of schizophrenia is already present during childhood. However, most of these studies were done in adult humans or animals. The present study therefore investigated age-related changes of behavior in an innovative version of the Open Field Maze following perinatal chronic administration of the noncompetitive N-methyl-D-aspartate antagonist MK-801. The waxing and waning pattern of locomotor activity during aging with a peak around the age of 90 days was profoundly attenuated by MK-801. MK-801 further reduced locomotion and body weight in mature rats. However, it increased explorative behavior in immature rats. Therefore, the chronic perinatal administration of MK-801 seems to be a model to investigate the pathological mechanisms of psychomotor retardation and body weight dysregulation in schizophrenia.

Knockout of c-Jun N-terminal kinases 1, 2 or 3 isoforms induces behavioural changes.

c-Jun N-terminal kinases (JNKs) are central and ubiquitous mediators of cellular signaling for both physiogical-regenerative and pathological-apoptotic processes. Their impact on degeneration or inflammation is well documented, but so far little is known about their roles in higher brain functions. The more, the contribution of individual JNK isoforms remains obscure so far. Here we have tested the behaviour of JNK1, JNK2 and JNK3 knockout (ko) mice in elevated plus maze (EPM), open field (OF), novel object recognition memory (NORM) test and Morris water maze (MWM). Compared with wild type C57BL/6N mice JNK ko mice revealed significant differences. Taken together the data on anxiety, exploration and learning indicate that JNK1 ko mice displayed a stronger explorative behaviour and that knockout of JNK2 or JNK3 showed a tendency of behaviour opposite to that of JNK1 ko mice. This pattern reminds of the impact of individual JNK ko on neurodegeneration. This is the first comparative study on the impact of individual JNK ko on behavioural parameters.

Effects of daytime naps on procedural and declarative memory in patients with schizophrenia.

Sleep has been identified as a state that optimizes the consolidation of newly acquired information in memory. Straight memory deficits and sleep disturbances are well-known in patients with schizophrenia. This study tested the hypothesis that patients with schizophrenia have a deficit in procedural and declarative memory consolidation after a short midday nap when compared to healthy controls and patients with remitted to moderate major depression. Following a normal night's sleep, 22 healthy subjects, 20 patients with major depression and 21 patients with schizophrenia were studied in a napping and wake condition in a random-order cross-over design, early in the afternoon. To test declarative memory, the Rey-Osterrieth Complex Figure Test respectively the Taylor Complex Figure Test and, for procedural learning, a mirror tracing task were performed. The present study is the first to demonstrate significant differences between individuals with schizophrenia, depression and healthy matched controls with regard to measures of sleep and memory performance after a short period of daytime sleep (napping). In particular we found that a daytime nap of only about 40min led to improvement of declarative memory performance in all investigated groups, whereas no beneficial effect was seen on procedural performance in the group of medicated patients with schizophrenia in contrast to healthy controls and patients with remitted to moderate major depression.

Are there age-related effects in antisocial personality disorders and psychopathy?

We investigated possible age differences in the frequency of antisocial personality disorder (ASPD), and of psychopathy according to Hare's criteria and its constituent features: In a cross-sectional study 226 male violent offenders, detained in adult or youth custody, were investigated using the Psychopathy Checklist Screening Version (PCL:SV) and the SCID II Interview. Their ages ranged from 18 to 59 years. Total PCL:SV score was negatively correlated with age. ANOVA showed that total PCL scores for three age groups differed significantly. Both effects were due entirely to Factor 2 of the PCL. Factor 1 was not related to age. The frequency of ASPD was also lower among older prisoners. The relationship with age was similar to that of PCL:SV Factor 2. These results point to age-related effects in psychopathy and suggest that different aspects of psychopathy follow different developmental courses. The results of our group comparison suggest that the different subfacets of psychopathy are not stable over time to the same extent. In order to make statements about the course of intraindividual development, however, longitudinal studies would be required.

The relationship between DSM-IV cluster B personality disorders and psychopathy according to Hare's criteria: clarification and resolution of previous contradictions.

This study examines the relationship between DSM-cluster B personality disorders (PDs) and psychopaths according to Hare's criteria as detected by the Psychopathy Checklist (PCL:SV) in 299 violent offenders. To clarify some contradictions among several previous studies on this issue, individual cluster B PDs were looked at alone, excluding any cases of comorbidity with other PDs of this cluster. We found highly significant relationships between antisocial and borderline PD and Factor II of the PCL and a highly significant correlation between narcissistic PD and Factor I of the PCL. These results were to be expected from the theoretical basis of the development of the PCL and provide a contribution to the construct validity of the PCL, which until now has not been validated on such a large sample in Germany.

Effects of short- and long-term pulsed radiofrequency electromagnetic fields on night sleep and cognitive functions in healthy subjects.

There has been wide public discussion on whether the electromagnetic fields of mobile telephones and their base stations affect human sleep or cognitive functioning. As there is evidence for learning and memory-consolidating effects of sleep and particularly of REM sleep, disturbance of sleep by radiofrequency electromagnetic fields might also impair cognitive functions. Previously realized sleep studies yielded inconsistent results regarding short-term exposure. Moreover, data are lacking on the effect that short- and long-term exposure might have on sleep as well as on cognitive functions. Therefore, 10 healthy young male subjects were included and nocturnal sleep was recorded during eight consecutive nights. In the second, third, and last night, we investigated polysomnographic night sleep and cognitive functions. After the adaptation and baseline nights, the participants were exposed to a defined radiofrequency electromagnetic field during the following six nights. We analyzed polysomnographic night sleep according to Rechtschaffen and Kales [1968, Manual of Standardized Terminology, Techniques and Scoring System for Sleep of Human Subjects] as well as by power spectra and correlation dimension. Cognitive functions were investigated by an array of neuropsychological tests. Data analysis was done by comparing the baseline night with the first and last exposure night and the first two sleep cycles of the respective nights. We did not find significant effects, either on conventional sleep parameters or on power spectra and correlation dimension, nor were there any significant effects on cognitive functions. With our results, we are unable to reveal either short-term or cumulative long-term effects of radiofrequency electromagnetic fields on night sleep and cognitive functions in healthy young male subjects.

Sleep in major depression: relation to memory performance and outcome after interpersonal psychotherapy.

BACKGROUND: Earlier findings suggest both a link between sleep and memory consolidation and a relationship between abnormal sleep at baseline and poor treatment outcome in major depression after interpersonal psychotherapy (IPT). METHODS: Pre-treatment polysomnography was examined in 32 patients with a major depressive episode (mean age = 39.5 years, 20 women). Declarative memory was tested by the Rey-Osterrieth Complex Figure Test and a paired associative word list and procedural learning was assessed by a mirror tracing skill. All patients were treated with IPT according to the manual and did not receive any antidepressant medication. Twenty-three patients took part in a minimum of 12 sessions of IPT. Remission was defined as 2 consecutive weeks with a score <8 on the Hamilton Rating Scale of Depression. RESULTS: Declarative visual memory performance was associated with total sleep time and total amount of rapid eye movement sleep. In IPT remitters (n = 14), there was a trend towards a decrease in rapid eye movement density (first period) and a significant decrease in delta power in pre-treatment sleep in comparison to non-remitters (n = 9). Treatment outcome after IPT was also associated with declarative memory performance at baseline (as a trend). CONCLUSIONS: Further indications of a role of sleep in memory processes and of the importance of specific sleep parameters as markers for a positive treatment response to psychotherapy were found.

[In-patient treatment concept for acute crises of borderline patients on the basis of dialectical-behavioral therapy]. / Stationäres Interventionskonzept auf Basis der Dialektisch-Behavioralen Therapie zur Behandlung akuter Krisen bei Borderline-Patienten.

Whereas the Dialectical-Behavioral Therapy (DBT) by Marsha Linehan for chronic suicidal female patients with borderline personality disorder has also been successfully implemented for in-patient treatment, the management of recurrent crisis has continued to be a significant challenge especially for hospitals taking part in the mandatory health coverage. Therefore, we have established an acute, short-term crisis treatment concept for both genders on a locked unit which takes the different circumstances of acute crisis into account. It ought to be used under the conditions of acute psychiatry and at the same time meet psychotherapeutic demands of borderline treatment. The treatment basis is DBT. The treatment focus is to improve stress-tolerance, especially by skills-training, regarding the individual situation and problems at time of admittance. The in-patient treatment should not take more than three weeks. The short-time aim is a fast reintegration in ambulant treatment or motivation for a specific therapy as DBT, middle- and long-time aims are the improvement of the ability to cope with crises and to demand less in-patient care.