Expression of G protein-coupled receptor 19 in human lung cancer cells is triggered by entry into S phase and supports G2/M cell cycle progression

G Protein-gekoppelte Rezeptoren (GPCRs) stellen die größte Familie zellulärer Oberflächenrezeptoren dar. Jedoch sind nur ein Teil der mehr als 800 Rezeptoren in solchem Ausmaß charakterisiert, dass deren physiologische Rolle gut verstanden wird. Tatsächlich werden in der heutigen Pharmakotherapie lediglich etwa 60 Vertreter dieser Rezeptoren mit einer Vielzahl von Substanzen angegriffen, welche etwa 30% aller verfügbaren Medikamente ausmachen. Es ist schon lange bekannt, dass GPCRs eine ungewöhnliche Expression in Krebszellen aufweisen können. An vielen Stellen der Karzinogenese tragen sonst normale, physiologische Funktionen von GPCRs mutmaßlich zur Krebsprogression bei, wenn diese unkontrolliert und missbräuchlich in einer Zelle verwendet werden. Das Hauptziel dieser Arbeit besteht darin, GPCRs vermehrt in Bezug zu malignen Erkrankungen zu stellen. Konkret wurde nach GPCRs gesucht, die eine hohe Expression im kleinzelligen Lungenkarzinom aufweisen. Hierbei zeigte sich, dass die den G Protein-gekoppelten Rezeptor 19 (GPR19) kodierende mRNA in Proben von Patienten mit kleinzelligem Lungenkarzinom verglichen mit Proben anderer Lungenkarzinome und von untransformiertem Lungengewebe überexprimiert war. Desweiteren deuten einige Beobachtungen darauf hin, dass eine Gpr19 Überexpression in von humanen Lungenkarzinomen abgeleiteten Zelllinien diesen einen spezifischen Vorteil im Durchschreiten des Zellzykluses bietet. Die Abschwächung der Gpr19 Genexpression mittels RNA Interferenz hemmte Lungenkarzinomszelllinien in ihrer Proliferation und führte letztlich zu deren Zelltod. Das Fortschreiten des Zellzykluses über die G2/M Phase hinweg war gestört, was mit einer erhöhten Präsenz an für diese Phasen charakteristischen Proteinmarkern (Zyklin B1, phosphoryliertes Histon H3) einherging. In den untersuchten Lungenkarzinomszelllinien war die Expression der Gpr19 mRNA abhängig vom Zellzyklus. Nachdem die Zellen in verschiedenen Phasen des Zellzykluses zunächst angehalten und danach aus dem Arrest entlassen wurden, konnte ein Höchststand der Gpr19 Expression während der S Phase konstatiert werden. Zudem zeigte sich mit Hilfe von Chromatin-Immunpräzipitation, dass die Gpr19 mRNA Expression in diesen Zelllinien anscheinend unter der Kontrolle von Transkriptionsfaktoren der E2F Familie steht. Mitglieder dieser Familie sind bekannt dafür, dass sie die Genexpression vieler am Zellzyklus beteiligter Gene kontrollieren. So konnte die Gpr19 Promotorregion bei der Anwendung von gegen E2F-1 bis 4 gerichteten Antikörpern nachgewiesen werden. Und die Zerstörung mutmaßlicher E2F Bindestellen in der Gpr19 Promotorregion schwächte die Expression eines Luciferase Reporterkonstrukts ab. Diese Arbeit offenbart zum ersten Mal die vom Zellzyklus abhängige mRNA Expression eines Vertreters der GPCR Familie, GPR19. Desweiteren stellt sie eine funktionelle Verbindung zwischen dieser differenziellen Expression, einem ungehinderten Durchschreiten des Zellzykluses und der Proliferation von humanen Lungenkarzinomszelllinien her, wenn diese eine erhöhte Gpr19 Grundexpression aufweisen. Folglich validieren diese Daten GPR19 als mögliches Angriffsmolekül bei einer Überexpression im Lungenkarzinom.G protein-coupled receptors (GPCRs) represent the largest family of cell surface receptors. However, only a fraction of the more than 800 receptors have been characterized to an extent that their physiological role is reasonably well understood. In fact, current pharmacotherapy only addresses some 60 receptors with a large collection of compounds that represent about 30% of the available drugs. It has long been known that GPCRs are subject to illegitimate expression in cancer cells. Presumably, hijacking the normal physiological functions of GPCRs contributes to all biological capabilities acquired during the multistep development of human cancers. With the goal of linking G protein-coupled receptors to malignant diseases, GPCRs were searched for that revealed high expression levels in small cell lung cancer (SCLC): The mRNA encoding orphan G protein-coupled receptor 19 (GPR19) was found to be frequently overexpressed in tissue samples obtained from patients with SCLC in contrast to samples derived from non-SCLC or normal lung. Several observations indicate that overexpression of Gpr19 confers a specific advantage to human lung cancer-derived cells regarding the transition through the cell cycle. Knockdown of Gpr19 mRNA by RNA interference reduced cell growth of human lung cancer cell lines and led to cell death. Cell cycle progression through G2/M phase was impaired and this was associated with increased protein levels of cyclin B1 and phosphorylated histone H3. Gpr19 exhibited a cell cycle-dependent expression pattern in lung cancer cell lines. When cell cycle distribution profiles of cells released from cell cycle arrest were related to Gpr19 mRNA levels, a peak Gpr19 expression was detected during S phase. The control of Gpr19 expression by E2F transcription factors, which drive gene expression of many genes important for cell cycle progression, was verified by chromatin immunoprecipitation: Antibodies directed against E2F-1 to E2F-4 allowed for the recovery of the Gpr19 promoter in lung cancer-derived cell lines. E2F binding site deletion in the Gpr19 promoter resulted in diminished luciferase reporter gene expression. This is the first example of a G protein-coupled receptor showing cell cycle phase specific mRNA expression. Further results point to a functional link between the cell cycle-dependent expression of Gpr19, a correct cell cycle passage, and cellular proliferation in human lung cancer-derived cells that exhibit higher than normal basal levels of Gpr19 mRNA. Hence, the data also validate GPR19 as a candidate target when overexpressed in lung cancer

The Diffusion of Organizations: The Role of Foreign Aid

Kühl S. The Diffusion of Organizations: The Role of Foreign Aid. In: Holzer B, Kastner F, Werron T, eds. From Globalization to World Society. Neo-Institutional and Systems-Theoretical Perspectives. Routledge advances in sociology. Vol 131. New York/London: Routledge; 2015: 258-278

Relapse Prevention Intervention after Suicidal Event (RISE): Feasibility study of a psychotherapeutic short-term program for inpatients after a recent suicide attempt

Recent research suggests that treating only mental disorders may not be sufficient to reduce the risk for future suicidal behavior in patients with a suicide attempt(s). It is therefore necessary to pay special therapeutic attention to past suicidal acts. Thus, the newly developed RISE (Relapse Prevention Intervention after Suicidal Event) program was built on the most effective components of existing psychotherapeutic and psychosocial interventions according to our current meta-analysis. The RISE program consists of five individual sessions designed for the acute psychiatric inpatient setting. The main goals of the treatment are to decrease future suicidal events and to improve patients' ability to cope with future suicidal crises. In the present study, feasibility and acceptance of the RISE program were investigated as well as its clinical effects on suicidal ideations, mental pain, self-efficacy and depressive symptoms. We recruited a sample of 27 inpatients of the Department of Psychiatry and Psychotherapy, University Hospital Jena, Germany. The final sample consisted of 20 patients hospitalized for a recent suicide attempt, including 60 percent of multiple attempters. The data collection included a structured interview and a comprehensive battery of questionnaires to evaluate the feasibility and acceptance of the RISE program as well as associated changes in clinical symptoms. A follow-up examination was carried out after 6 months. Considering the low dropout rate and the overall positive evaluation, the RISE program was highly accepted in a sample of severely impaired patients. The present study also demonstrated that the levels of suicidal ideations, mental pain, depressive symptoms, and hopelessness decreased significantly after RISE. Since all of these clinical parameters are associated with the risk of future suicidal behavior, a potential suicide-preventive effect of the intervention can be inferred from the present findings. The positive results of the follow-up assessment after 6 months point in the same direction. In addition, RISE treatment increased self-efficacy in patients, which is an important contributor for better coping with future suicidal crises. Thus, present study demonstrate that RISE is a suitable therapy program for the treatment of patients at high risk for suicidal behavior in an acute inpatient setting

Did female prisoners with mental disorders receive psychiatric treatment before imprisonment?

© 2015 Mundt et al.; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.BackgroundThroughout the world, high prevalence rates of mental disorders have been found in prison populations, especially in females. It has been suggested that these populations do not access psychiatric treatment. The aim of this study was to establish rates of psychiatric in- and outpatient treatments prior to imprisonment in female prisoners and to explore reasons for discontinuation of such treatments.Methods150 consecutively admitted female prisoners were interviewed in Berlin, Germany. Socio-demographic characteristics, mental disorders, and previous psychiatric in- and outpatient treatments were assessed by trained researchers. Open questions were used to explore reasons for ending previous psychiatric treatment.ResultsA vast majority of 99 prisoners (66%; 95% CI: 58¿73) of the total sample reported that they had previously been in psychiatric treatment, 80 (53%; 95 CI: 45¿61) in inpatient treatment, 62 (41%; 95 CI: 34¿49) in outpatient treatment and 42 (29%; 21¿39) in both in- and outpatient treatments. All prisoners with psychosis and 72% of the ones with any lifetime mental health disorder had been in previous treatment. The number of inpatient treatments and imprisonments were positively correlated (rho¿=¿0.27; p¿<¿0.01). Inpatient treatment was described as successfully completed by 56% (N¿=¿41) of those having given reasons for ending such treatment, whilst various reasons were reported for prematurely ending outpatient treatments.ConclusionThe data do not support the notion of a general `mental health treatment gap¿ in female prisoners. Although inpatient care is often successfully completed, repeated inpatient treatments are not linked with fewer imprisonments. Improved transition from inpatient to outpatient treatment and services that engage female prisoners to sustained outpatient treatments are needed

Prevalence Rates of Mental Disorders in Chilean Prisons

PMCID: PMC371883

Bond-disordered spin systems: Theory and application to doped high-Tc compounds

We examine the stability of magnetic order in a classical Heisenberg model with quenched random exchange couplings. This system represents the spin degrees of freedom in high-$T_\textrm{c}$ compounds with immobile dopants. Starting from a replica representation of the nonlinear $\sigma$-model, we perform a renormalization-group analysis. The importance of cumulants of the disorder distribution to arbitrarily high orders necessitates a functional renormalization scheme. From the renormalization flow equations we determine the magnetic correlation length numerically as a function of the impurity concentration and of temperature. From our analysis follows that two-dimensional layers can be magnetically ordered for arbitrarily strong but sufficiently diluted defects. We further consider the dimensional crossover in a stack of weakly coupled layers. The resulting phase diagram is compared with experimental data for La$_{2-x}$Sr$_x$CuO$_4$.Comment: 12 pages, 5 figure

The microaerophilic microbiota of de-novo paediatric inflammatory bowel disease: the BISCUIT study

&lt;p&gt;Introduction: Children presenting for the first time with inflammatory bowel disease (IBD) offer a unique opportunity to study aetiological agents before the confounders of treatment. Microaerophilic bacteria can exploit the ecological niche of the intestinal epithelium; Helicobacter and Campylobacter are previously implicated in IBD pathogenesis. We set out to study these and other microaerophilic bacteria in de-novo paediatric IBD.&lt;/p&gt; &lt;p&gt;Patients and Methods: 100 children undergoing colonoscopy were recruited including 44 treatment naïve de-novo IBD patients and 42 with normal colons. Colonic biopsies were subjected to microaerophilic culture with Gram-negative isolates then identified by sequencing. Biopsies were also PCR screened for the specific microaerophilic bacterial groups: Helicobacteraceae, Campylobacteraceae and Sutterella wadsworthensis.&lt;/p&gt; &lt;p&gt;Results: 129 Gram-negative microaerophilic bacterial isolates were identified from 10 genera. The most frequently cultured was S. wadsworthensis (32 distinct isolates). Unusual Campylobacter were isolated from 8 subjects (including 3 C. concisus, 1 C. curvus, 1 C. lari, 1 C. rectus, 3 C. showae). No Helicobacter were cultured. When comparing IBD vs. normal colon control by PCR the prevalence figures were not significantly different (Helicobacter 11% vs. 12%, p = 1.00; Campylobacter 75% vs. 76%, p = 1.00; S. wadsworthensis 82% vs. 71%, p = 0.312).&lt;/p&gt; &lt;p&gt;Conclusions: This study offers a comprehensive overview of the microaerophilic microbiota of the paediatric colon including at IBD onset. Campylobacter appear to be surprisingly common, are not more strongly associated with IBD and can be isolated from around 8% of paediatric colonic biopsies. S. wadsworthensis appears to be a common commensal. Helicobacter species are relatively rare in the paediatric colon.&lt;/p&gt