Evaluation of a new body-focused group therapy versus a guided self-help group program for adults with psychogenic non-epileptic seizures (PNES): a pilot randomized controlled feasibility study

Objective: Psychogenic non-epileptic seizures (PNES), a common phenomenon in neurological settings, are regarded as a paroxysmal type of functional neurological disorder (FND). In a substantial proportion, PNES are disabling with poor long-term outcomes and high economic costs. Despite the clinical and financial consequences of PNES, there is still a lack of controlled clinical trials on the treatment of this challenging disorder. The study aims to evaluate the feasibility and collect first evidence of the efficacy of a group based-intervention in PNES-patients. Methods: A pilot randomized controlled feasibility study with a parallel-group design was performed in adult outpatients with PNES to evaluate a new body-focused group therapy (CORDIS) versus guided self-help groups. Self-assessment of dissociation (Dissociation Experience Scale-DES-20) and seizure severity (Liverpool Seizure Severity Scale-LSSS) were assessed two weeks before and two weeks after the treatment intervention and also six months after treatment as primary outcome parameters. Results: A total of 53 patients were recruited from a specialized outpatient clinic, and out of those, 29 patients completed either the body-focused group therapy program (n = 15) or a guided self-help group (SHG) therapy (n = 14). When analyzing the ITT sample (n = 22 CORDIS group, n = 20 SHG), both groups showed an effect on seizure severity and level of dissociation. In the per protocol sample (n = 13 CORDIS group, n = 12 SHG), CORDIS was superior to the self-help group for reducing seizure severity 6 months after the treatment. Significance: CORDIS is a newly developed body-focused group therapy program for adults with PNES. Further studies should include a multicentric design with a higher number of participants

Identification of single-point mutations in mycobacterial 16S rRNA sequences by confocal single-molecule fluorescence spectroscopy

We demonstrate the specific identification of single nucleotide polymorphism (SNP) responsible for rifampicin resistance of Mycobacterium tuberculosis applying fluorescently labeled DNA-hairpin structures (smart probes) in combination with single-molecule fluorescence spectroscopy. Smart probes are singly labeled hairpin-shaped oligonucleotides bearing a fluorescent dye at the 5′ end that is quenched by guanosine residues in the complementary stem. Upon hybridization to target sequences, a conformational change occurs, reflected in a strong increase in fluorescence intensity. An excess of unlabeled (‘cold’) oligonucleotides was used to prevent the formation of secondary structures in the target sequence and thus facilitates hybridization of smart probes. Applying standard ensemble fluorescence spectroscopy we demonstrate the identification of SNPs in PCR amplicons of mycobacterial rpoB gene fragments with a detection sensitivity of 10(−8) M. To increase the detection sensitivity, confocal fluorescence microscopy was used to observe fluorescence bursts of individual smart probes freely diffusing through the detection volume. By measuring burst size, burst duration and fluorescence lifetime for each fluorescence burst the discrimination accuracy between closed and open (hybridized) smart probes could be substantially increased. The developed technique enables the identification of SNPs in 10(−11) M solutions of PCR amplicons from M.tuberculosis in only 100 s

Use of Autoreactive Antibodies in Blood of Patients with Pancreatic Intraductal Papillary Mucinous Neoplasms (IPMN) for Grade Distinction and Detection of Malignancy

(1) Background: A reliable non-invasive distinction between low- and high-risk pancreatic intraductal papillary mucinous neoplasms (IPMN) is needed to effectively detect IPMN with malignant potential. This would improve preventative care and reduce the risk of developing pancreatic cancer and overtreatment. The present study aimed at exploring the presence of autoreactive antibodies in the blood of patients with IPMN of various grades of dysplasia. (2) Methods: A single-center cohort was studied composed of 378 serum samples from patients with low-grade IPMN (n = 91), high-grade IPMN (n = 66), IPMN with associated invasive cancer (n = 30), pancreatic ductal adenocarcinoma (PDAC) stages T1 (n = 24) and T2 (n = 113), and healthy controls (n = 54). A 249 full-length recombinant human protein microarray was used for profiling the serum samples. (3) Results: 14 proteins were identified as potential biomarkers for grade distinction in IPMN, yielding high specificity but mediocre sensitivity. (4) Conclusions: The identified autoantibodies are potential biomarkers that may assist in the detection of malignancy in IPMN patients

Stratification of pancreatic tissue samples for molecular studies: RNA-based cellular annotation procedure

Abstract Background/objectives Meaningful profiling of pancreatic cancer samples is particularly challenging due to their complex cellular composition. Beyond tumor cells, surgical biopsies contain desmoplastic stroma with infiltrating inflammatory cells, adjacent normal parenchyma, and "non-pancreatic tissues". The risk of misinterpretation rises when the heterogeneous cancer tissues are sub-divided into smaller fragments for multiple analytic procedures. Pre-analytic histological evaluation is the best option to characterize pancreatic tissue samples. Our aim was to develop a complement or alternative procedure to determine the cellular composition of pancreatic cancerous biopsies, basing on intra-analytic molecular annotation. A standard process for sample stratification at a molecular level does not yet exist. Particularly in the case of retrospective or data depository-based studies, when hematoxylin-eosin stained sections are not available, it supports the correct interpretation of expression profiles. Methods A five-gene transcriptional signature ( RNA CellStrat) was defined that allows cell type-specific stratification of pancreatic tissues. Testing biopsy material from biobanks with this procedure demonstrated high correspondence of molecular (qRT-PCR and microarray) and histologic (hematoxylin-eosin stain) evaluations. Results Notably, about a quarter of randomly selected samples (tissue fragments) were exposed as inappropriate for subsequent clinico-pathological interpretation. Conclusions Via immediate intra-analytical procedure, our RNA-based stratification RNA CellStrat increases the accuracy and reliability of the conclusions drawn from diagnostic and prognostic molecular information

Strong Association of a Common Dihydropyrimidine Dehydrogenase Gene Polymorphism with Fluoropyrimidine-Related Toxicity in Cancer Patients

variations associated with enhanced drug toxicity. = 0.001; the attributable risk was 56.9%. Comparing tumor-type matched sets of samples, correlation of c.496A>G with toxicity was particularly present in patients with gastroesophageal and breast cancer, but did not reach significance in patients with colorectal malignancies. polymorphism strongly contributes to the occurrence of fluoropyrimidine-related drug adverse effects. Carriers of this variant could benefit from individual dose adjustment of the fluoropyrimidine drug or alternate therapies

The introduction of benzodiazepines at the Charité Nervenklinik from 1962-75

Einleitung Zu Beginn der 1960er-Jahre kamen in der westlichen Welt die Benzodiazepine auf den Markt. Schnell wurden sie dort zu den meistverkauften Medikamenten überhaupt. In der DDR waren sie jedoch aufgrund restriktiver Importbestimmungen und ideologischer Bedenken zunächst nicht verfügbar. Die Voraussetzung für einen Import oder die nationale Eigenproduktion war ein positiver Bescheid zentraler, durch das Ministerium für Gesundheitswesen gelenkter Ausschüsse und Gremien. Dieser wurde schließlich getroffen und man begann die Substanzen zu analysieren und ab Ende 1967 zu produzieren. So konnten in der Nervenklinik der Berliner Charité erste klinische Erfahrungen mit den nun breit verfügbaren Benzodiazepinen gesammelt werden. Zielstellung In einem mehrschrittigen Vorgehen habe ich die Jahre vor und nach der Verfügbarkeit der Medikamente unter besonderer Berücksichtigung der Charité von 1962-1975 untersucht. Ziel war es dabei erstens, die ärztliche Anwendung der neuen Beruhigungsmittel zu charakterisieren. Zweitens sollte die Sensibilität der Kliniker für das Abhängigkeitspotential der Medikamente beurteilt werden. Drittens fragte ich nach dem Einfluss von DDR-Bürgern auf die Entscheidung zur nationalen Produktion von Benzodiazepinen. Methodik Meine Untersuchung basiert auf der Auswertung von Kranken- und Verwaltungsakten der Nervenklinik. Nach einer explorativen Sichtung wurden folgende Kriterien für eine Auswertung der Krankenakten festgelegt: 1. Die Anwendung von sedierenden Medikamenten während des stationären Aufenthalts 2. Der dokumentierte Gebrauch von Beruhigungsmitteln vor der stationären Aufnahme 3. Die Diagnose Medikamentenmissbrauch oder -abhängigkeit. Es wurden drei Zufallsstichproben von insgesamt 732 Akten aus den Jahrgängen 1962, 1968 und 1975 ausgewertet. Bei allen Akten, welche die genannten Kriterien erfüllten, wurden Variablen wie das Geschlecht, der soziale Status der Patienten, die Diagnosen und die Anzahl der Behandlungstage mit einem Beruhigungsmittel bestimmt und die Häufigkeiten für die Publikationen selektiv ausgewertet. Zusätzlich erfolgte eine qualitative Analyse, welche etwa eine Beschreibung der Indikationsstellung und die Beurteilung als Medikamentenabhängigkeit ermöglichte. Weiterhin wurde die zeitgenössische wissenschaftliche Literatur und Petitionen von DDR-Bürgern an das Ministerium für Gesundheitswesen der DDR ausgewertet. Ergebnisse und Schlussfolgerungen In einer ersten Veröffentlichung stellte ich die ärztliche Anwendung der Benzodiazepine dar. Diese orientierte sich stark an den verbreiteten Barbituraten. Nur unzureichend konnten die neuen Präparate in das psychopathologische Forschungsprogramm der Klinik integriert werden und wurden zunächst sehr zurückhaltend eingesetzt. In einer weiteren Arbeit untersuchte ich die Sensibilisierung für das Suchtpotential der Substanzen. Überraschender Weise konnte diesbezüglich eine deutlich zunehmende Akzeptanz für die regelmäßige Einnahme der Präparate in den 1970er-Jahren beobachtet werden. Schließlich erörterte ich in einem dritten Aufsatz, wie die DDR-Bürger durch Petitionen an das zuständige Ministerium die politische Entscheidung zur Produktion von Benzodiazepinen beförderten.Introduction Benzodiazepines were launched on western markets at the beginning of the 1960s and rapidly became blockbuster drugs. However, these drugs were not available in the GDR due to import restrictions and ideological doubts. The main prerequisite for importation or national production was a favourable decision by centralized panels controlled by the Ministry of Health. Once this decision was eventually taken, chemical analysis began and national production was underway by the end of 1967. Clinicians at the Berlin Charité Nervenklinik were then able to gain their first clinical experiences with the now widely available benzodiazepines. Objectives I observed the years before and after introduction of the benzodiazepines with a special focus on the Charité between 1962-1975. I first aimed to characterize the application of the new sedatives in clinical practice. Secondly, I observed the perception of their addictive potential among clinicians. I thirdly asked how GDR citizens influenced drug policy to achieve the national production of benzodiazepines. Methods My study is based on an examination of the Charité-Nervenklinik’s patient and administrative files. After an explorative survey, the following criteria for further investigation were defined: 1. the application of sedatives in the medications during hospitalisation; 2. the documented usage of sedatives before admittance to the hospital; 3. diagnosis of drug abuse or addiction. Three random samples from a total of 732 patient files were taken from the volumes 1962, 1968 and 1975. Of all the files that met the above- mentioned criteria, variables such as sex, social status, diagnosis and the duration of application of a sedative were included and frequencies were determined specifically regarding the scope of the publication. Additionally, a qualitative analysis allowed for descriptions such as indication for treatment and the evaluation of abusive behaviour. Furthermore, the contemporary scientific literature and petitions of GDR citizens to the Ministry of Health were examined. Results and conclusions In a first article I portrayed the medical application of the benzodiazepines in clinical practice. It was closely related to barbiturates, which had been in widespread use for a considerable period of time. The drugs` application could not be matched with the psychopathological research interests at the Nervenklinik and were in limited use. In a second work, I examined the perception of the drugs`addictive potential among clinicians. Surprisingly, an increasing tolerance towards regular intake of benzodiazepines was observed in the 1970s. And finally, in a third published work, I discussed how GDR citizens used petitions to the Ministry of Health to push for a political decision on the national production of benzodiazepines

An open-label pilot study of the efficacy and safety of anakinra in patients with psoriatic arthritis refractory to or intolerant of methotrexate (MTX)

Only limited data have been published about the therapeutic use of anakinra in patients with psoriatic arthritis. We undertook this study to evaluate the efficacy and safety of anakinra in patients with active psoriatic arthritis. In a prospective open-label single-center study, 20 patients were treated with 100 mg anakinra everyday either alone or in combination with ongoing methotrexate over 6 months. Safety and efficacy was evaluated using Psoriasis Arthritis Response Criteria (PsARC), Disease Activity Score (DAS) 28, American College of Rheumatology (ACR), European League Against Rheumatism (EULAR), Psoriasis Area and Severity Index Score, Dactylitis Score and Health Assessment Questionnaire (HAQ), and the C-reactive protein, and erythrocyte sedimentation rate. Of the 20 patients enrolled, six completed 24 weeks, 18 completed 12 weeks, and 19 completed 4 weeks of treatment. Early-treatment termination was mainly due to inefficacy (13 patients) and only one drop-out occurred because of an unrelated adverse event. Six patients fulfilled continuously the PsARC until week 24. A moderate EULAR response was achieved by four patients and a good EULAR response by three patients in week 24. Five patients reached ACR 20, four patients ACR 50, and two patients ACR 70 in week 24. HAQ improved slightly throughout the study (n = 19, mean (SD); baseline, 1.127 (0.671); week 24, 1.055 (0.812)) just as DAS 28 (n = 16; baseline, 4.7(1.5); week 24, 4.0(2.0)). Only nine patients showed skin manifestations affecting > 3% of their body surface area which improved in two, worsened in four, stabilized in two patients, and newly evolved in one patient. Adverse events were mainly mild (95%). Fifteen (75%) patients showed injection site reactions. No serious infections occurred. Anakinra was well tolerated with no occurrence of serious drug-associated adverse events and lead to improvement of signs and symptoms in nine out of 19 patients, therefore providing a potential therapeutic option in patients with active psoriatic arthritis

Genes and Proteins Differentially Expressed during In Vitro Malignant Transformation of Bovine Pancreatic Duct Cells

Pancreatic carcinoma has an extremely bad prognosis due to lack of early diagnostic markers and lack of effective therapeutic strategies. Recently, we have established an in vitro model recapitulating the first steps in the carcinogenesis of the pancreas. SV40 large T antigen-immortalized bovine pancreatic duct cells formed intrapancreatic adenocarcinoma tumors on k-ras(mut) transfection after orthotopic injection in the nude mouse pancreas. Here we identified genes and proteins differentially expressed in the course of malignant transformation using reciprocal suppression subtractive hybridization and 2D gel electrophoresis and mass spectrometry, respectively. We identified 34 differentially expressed genes, expressed sequence tags, and 15 unique proteins. Differential expression was verified for some of the genes or proteins in samples from pancreatic carcinoma. Among these genes and proteins, the majority had already been described either to be influenced by a mutated ras or to be differentially expressed in pancreatic adenocarcinoma, thus proving the feasibility of our model. Other genes and proteins (e.g., BBC1, GLTSCR2, and rhoGDIα), up to now, have not been implicated in pancreatic tumor development. Thus, we were able to establish an in vitro model of pancreatic carcinogenesis, which enabled us to identify genes and proteins differentially expressed during the early steps of malignant transformation

Establishment and characterization of a highly tumourigenic and cancer stem cell enriched pancreatic cancer cell line as a well defined model system.

Standard cancer cell lines do not model the intratumoural heterogeneity situation sufficiently. Clonal selection leads to a homogeneous population of cells by genetic drift. Heterogeneity of tumour cells, however, is particularly critical for therapeutically relevant studies, since it is a prerequisite for acquiring drug resistance and reoccurrence of tumours. Here, we report the isolation of a highly tumourigenic primary pancreatic cancer cell line, called JoPaca-1 and its detailed characterization at multiple levels. Implantation of as few as 100 JoPaca-1 cells into immunodeficient mice gave rise to tumours that were histologically very similar to the primary tumour. The high heterogeneity of JoPaca-1 was reflected by diverse cell morphology and a substantial number of chromosomal aberrations. Comparative whole-genome sequencing of JoPaca-1 and BxPC-3 revealed mutations in genes frequently altered in pancreatic cancer. Exceptionally high expression of cancer stem cell markers and a high clonogenic potential in vitro and in vivo was observed. All of these attributes make this cell line an extremely valuable model to study the biology of and pharmaceutical effects on pancreatic cancer