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Introduction: Preclinical CT-guided radiotherapy platforms are increasingly used but the CT images are characterized by poor soft tissue contrast. The aim of this study was to develop a robust and accurate method of MRI-guided radiotherapy (MR-IGRT) delivery to abdominal targets in the mouse.
Methods: A multimodality cradle was developed for providing subject immobilisation and its performance was evaluated. Whilst CT was still used for dose calculations, target identification was based on MRI. Each step of the radiotherapy planning procedure was validated initially in vitro using BANG gel dosimeters. Subsequently, MR-IGRT of normal adrenal glands with a size-matched collimated beam was performed. Additionally, the SK-N-SH neuroblastoma xenograft model and the transgenic KPC model of pancreatic ductal adenocarcinoma were used to demonstrate the applicability of our methods for the accurate delivery of radiation to CT-invisible abdominal tumours.
Results: The BANG gel phantoms demonstrated a targeting efficiency error of 0.56 ± 0.18 mm. The in vivo stability tests of body motion during MR-IGRT and the associated cradle transfer showed that the residual body movements are within this MR-IGRT targeting error. Accurate MR-IGRT of the normal adrenal glands with a size-matched collimated beam was confirmed by γH2AX staining. Regression in tumour volume was observed almost immediately post MR-IGRT in the neuroblastoma model, further demonstrating accuracy of x-ray delivery. Finally, MR-IGRT in the KPC model facilitated precise contouring and comparison of different treatment plans and radiotherapy dose distributions not only to the intra-abdominal tumour but also to the organs at risk.
Conclusion: This is, to our knowledge, the first study to demonstrate preclinical MR-IGRT in intra-abdominal organs. The proposed MR-IGRT method presents a state-of-the-art solution to enabling robust, accurate and efficient targeting of extracranial organs in the mouse and can operate with a sufficiently high throughput to allow fractionated treatments to be given.
Glial cell line-derived neurotrophic factor (GDNF) is a ligand that activates, through co-receptor GDNF family receptor alpha-1 (GFRα1) and receptor tyrosine kinase “RET”, several signaling pathways crucial in the development and sustainment of multiple neuronal populations. We decided to study whether non-mammalian orthologs of these three proteins have conserved their function: can they activate the human counterparts? Using the baculovirus expression system, we expressed and purified Danio rerio RET, and its binding partners GFRα1 and GDNF, and Drosophila melanogaster RET and two isoforms of co-receptor GDNF receptor-like. Our results report high-level insect cell expression of post-translationally modified and dimerized zebrafish RET and its binding partners. We also found that zebrafish GFRα1 and GDNF are comparably active as mammalian cell-produced ones. We also report the first measurements of the affinity of the complex to RET in solution: at least for zebrafish, the Kd for GFRα1-GDNF binding RET is 5.9 μM. Surprisingly, we also found that zebrafish GDNF as well as zebrafish GFRα1 robustly activated human RET signaling and promoted the survival of cultured mouse dopaminergic neurons with comparable efficiency to mammalian GDNF, unlike E. coli-produced human proteins. These results contradict previous studies suggesting that mammalian GFRα1 and GDNF cannot bind and activate non-mammalian RET and vice versa.
Compact stars can be treated as the ultimate laboratories for testing theories of dense matter. They are not only extremely dense objects, but they are known to be associated with strong magnetic fields, fast rotation and, in certain cases, with very high temperatures. Here, we present several different approaches to model numerically the signatures and properties of these stars, namely:
•The effects of strong magnetic fields on hybrid stars by using a fully general relativistic approach. We solved the coupled Maxwell-Einstein equations in a self-consistent way, taking into consideration the anisotropy of the energy-momentum tensor due purely to the magnetic field, magnetic field effects on equation of state and the interaction between matter and the magnetic field (magnetization). We showed that the effects of the magnetization and the magnetic field on the equation of state for matter do not play an important role on global properties of neutron stars (only the pure magnetic _eld contribution does). In addition, the magnetic field breaks the spherical symmetry of stars, inducing major changes in the populated degrees of freedom inside these objects and, potentially, converting a hybrid star into a hadronic star over time.
•The effects of magnetic fields and rotation on the structure and composition of proto-neutron stars. We found that the magnetic field not only deforms these stars, but also significantly alters the number of trapped neutrinos in the stellar interior, together with the strangeness content and temperature in each evolution stage from a hot proto-neutron star to a cold neutron star.
•The influence of the quark-hadron phase transitions in neutron stars. In particular, previous calculations have shown that fast rotating neutron stars, when subjected to a quark-hadron phase transition in their interiors, could give rise to the backbending phenomenon characterized by a spin-up era. In this work, we obtained the interesting backbending phenomenon for fast spinning neutron stars. More importantly, we showed that a magnetic field, which is assumed to be axisymmetric and poloidal, can also be enhanced due to the phase transition from normal hadronic matter to quark matter on highly magnetized neutron stars. Therefore, in parallel to the spin-up era, classes of neutron stars endowed with strong magnetic fields may go through a `magnetic-up era' in their lives.
•Finally, we were also able to calculate super-heavy white dwarfs in the presence of strong magnetic fields. White dwarfs are the progenitors of supernova Type Ia explosions and they are widely used as candles to show that the Universe is expanding and accelerating. However, observations of ultraluminous supernovae have suggested that the progenitor of such an explosion should be a white dwarf with mass above the well-known Chandrasekhar limit ~ 1.4 M. In corroboration with other works, but by using a fully general relativistic framework, we obtained also strongly magnetized white dwarfs with masses M ~ 2:0 M.
For a chaotic system pairs of initially close-by trajectories become eventually fully uncorrelated on the attracting set. This process of decorrelation can split into an initial exponential decrease and a subsequent diffusive process on the chaotic attractor causing the final loss of predictability. Both processes can be either of the same or of very different time scales. In the latter case the two trajectories linger within a finite but small distance (with respect to the overall extent of the attractor) for exceedingly long times and remain partially predictable. Standard tests for chaos widely use inter-orbital correlations as an indicator. However, testing partially predictable chaos yields mostly ambiguous results, as this type of chaos is characterized by attractors of fractally broadened braids. For a resolution we introduce a novel 0-1 indicator for chaos based on the cross-distance scaling of pairs of initially close trajectories. This test robustly discriminates chaos, including partially predictable chaos, from laminar flow. Additionally using the finite time cross-correlation of pairs of initially close trajectories, we are able to identify laminar flow as well as strong and partially predictable chaos in a 0-1 manner solely from the properties of pairs of trajectories.
Background: The Objective Structured Clinical Examination (OSCE) is increasingly used at medical schools to assess practical competencies. To compare the outcomes of students at different medical schools, we introduced standardized OSCE stations with identical checklists.
Methods: We investigated examiner bias at standardized OSCE stations for knee- and shoulder-joint examinations, which were implemented into the surgical OSCE at five different medical schools. The checklists for the assessment consisted of part A for knowledge and performance of the skill and part B for communication and interaction with the patient. At each medical faculty, one reference examiner also scored independently to the local examiner. The scores from both examiners were compared and analysed for inter-rater reliability and correlation with the level of clinical experience. Possible gender bias was also evaluated.
Results: In part A of the checklist, local examiners graded students higher compared to the reference examiner; in part B of the checklist, there was no trend to the findings. The inter-rater reliability was weak, and the scoring correlated only weakly with the examiner’s level of experience. Female examiners rated generally higher, but male examiners scored significantly higher if the examinee was female.
Conclusions: These findings of examiner effects, even in standardized situations, may influence outcome even when students perform equally well. Examiners need to be made aware of these biases prior to examining.
Parkinson's disease (PD) is a frequent neurodegenerative process in old age. Accumulation and aggregation of the lipid-binding SNARE complex component α-synuclein (SNCA) underlies this vulnerability and defines stages of disease progression. Determinants of SNCA levels and mechanisms of SNCA neurotoxicity have been intensely investigated. In view of the physiological roles of SNCA in blood to modulate vesicle release, we studied blood samples from a new large pedigree with SNCA gene duplication (PARK4 mutation) to identify effects of SNCA gain of function as potential disease biomarkers. Downregulation of complexin 1 (CPLX1) mRNA was correlated with genotype, but the expression of other Parkinson's disease genes was not. In global RNA-seq profiling of blood from presymptomatic PARK4 indviduals, bioinformatics detected significant upregulations for platelet activation, hemostasis, lipoproteins, endocytosis, lysosome, cytokine, Toll-like receptor signaling and extracellular pathways. In PARK4 platelets, stimulus-triggered degranulation was impaired. Strong SPP1, GZMH and PLTP mRNA upregulations were validated in PARK4. When analysing individuals with rapid eye movement sleep behavior disorder, the most specific known prodromal stage of general PD, only blood CPLX1 levels were altered. Validation experiments confirmed an inverse mutual regulation of SNCA and CPLX1 mRNA levels. In the 3′-UTR of the CPLX1 gene we identified a single nucleotide polymorphism that is significantly associated with PD risk. In summary, our data define CPLX1 as a PD risk factor and provide functional insights into the role and regulation of blood SNCA levels. The new blood biomarkers of PARK4 in this Turkish family might become useful for PD prediction.
Patient-reported outcomes (PROs) such as quality of life and work productivity are important for measuring patient's experience. We assessed PROs during and after treatment of hepatitis C virus (HCV) patients.Data were obtained from a phase 3 open label study of sofosbuvir and ribavirin (SOF + RBV) with and without interferon (IFN). Patients completed 4 PRO assessment instruments (SF-36, Functional Assessment of Chronic Illness Therapy-Fatigue, Chronic Liver Disease Questionnaire- HCV, Work Productivity and Activity-Specific Health Problem) before, during, and after treatment.A total of 533 patients with chronic HCV were enrolled; 28.9% treatment-naïve, 23.1% cirrhotic, 219 received IFN + SOF + RBV and 314 received IFN-free SOF + RBV. At baseline, there were no differences in PROs between the IFN-free and IFN-containing treatment arms (all P > 0.05). During treatment, patients receiving IFN + SOF + RBV had a substantial impairment in their PROs (up to -24.4% by treatment week 12, up to -8.3% at week 4 post-treatment). The PRO decrements seen in the SOF + RBV arm were smaller in magnitude (up to -7.1% by treatment week 12), and all returned to baseline or improved by post-treatment week 4. By 12 weeks after treatment cessation, patients who achieved sustained viral response-12 showed some improvement of PRO scores regardless of the regimen (up to +7.1%, P < 0.0001) or previous treatment experience. In multivariate analysis, the use of IFN was independently associated with lower PROs.IFN-based regimens have a profoundly negative impact to PROs. By contrast, the impact of RBV on these PROs is relatively modest. Achieving HCV cure is associated with improvement of most of the PRO scores.
Expression, perception and recognition of intense emotions in healthy and depressed individuals
(2017)
Die Fähigkeit die Gefühle anderer zu erkennen und einzuordnen ermöglicht es soziale Situationen richtig einzuschätzen und soziale Beziehungen aufzubauen. Da Emotionen also in unserem Leben eine wichtige Rolle spielen, kann eine Dysregulation der Emotionsverarbeitung auch zu elementaren Einschränkungen führen. Menschen, die unter depressiven Episoden leiden, durchleben beispielsweise regelmäßig Phasen intensiver und anhaltender Traurigkeit. Jedoch ist noch nicht vollständig erklärt, wie es zu dieser verzerrten Emotionswahrnehmung kommt. Diese Dissertation hatte deshalb das Ziel, den Ausdruck, die Wahrnehmung und das Erkennen extremer Emotionen genauer zu beleuchten.
In Studie 1 wurden der Ausdruck und das Erkennen extremer Emotionen untersucht.
Hierbei dienten aus dem Internet bezogene Videosequenzen von Kindern und Erwachsenen als Basis, in denen diese sich in Situationen befanden, die sie extrem negative oder extrem positive Emotionen durchleben ließen. Die Gesichtsausdrücke der Kinder und Erwachsenen wurden dann zum Zeitpunkt der stärksten emotionalen Erregung in ein Bild umgewandelt und von unabhängigen Ratern auf ihre Valenz und ihr Arousal eingeschätzt. Es wurde beobachtet, dass - entgegen der Vorhersage etablierter Emotionstheorien (z.B. Ekman, 1993) – Emotionen hoher positiver und negativer Intensität schwer auseinander zu halten sind. Tatsächlich wurden positive Emotionsausdrücke häufig als negativ eingeschätzt. Eine mögliche Erklärung dafür liefern Aragón und Kollegen (2015). Sie schätzen den Ausdruck negativer Emotionen in positiven Situationen als Emotionsregulationsstrategie ein, die dazu dient ein emotionales Equilibrium wieder herzustellen, das durch die überwältigenden positiven Emotionen aus dem Gleichgewicht gebracht wurde.
In Studie 2 und 3 wurde die Wahrnehmung negativer Emotionen bei depressiven Menschen im Vergleich zu gesunden Kontrollprobanden auf subjektiver und physiologischer Ebene untersucht. Hierbei wurde zunächst im Rahmen von Studie 2 untersucht, ob Parameter des autonomen Nervensystems (ANS) sich zwischen depressiven und gesunden Probanden unterscheiden. ANS-Parameter umfassten Hormone (Cortisol und DHEA), Herzratenvariabilität (HRV), Hautleitfähigkeit (GSR), Hauttemperatur (TEMP) und Atemfrequenz (RSP). Es konnten erhöhte DHEA-Werte, eine erhöhte Hauttemperatur und eine reduzierte Atemfrequenz in der Patientengruppe gefunden werden. Eine erhöhte Hauttemperatur korrelierte zudem mit der Ausprägung depressiver Symptome und der aktuellen Stimmung. Reduzierte HRV-Werte wurden hauptsächlich auf antidepressive Medikation zurückgeführt.
In Studie 3 wurde dann die Reaktion der Probanden auf emotionsevozierende Stimuli verschiedener Valenzkategorien (neutral, leicht negative, hoch negative) untersucht. Hierbei wurden sowohl physiologische Parameter (TEMP, HRV, GSR, RSP) als auch die subjektive Einschätzung der Stimuli bezüglich ihrer Valenz und ihres Arousal erhoben. Die Befunde bezüglich Hauttemperatur und HRV-Werte aus Studie 2 konnten in Studie 3 repliziert
werden. Zudem zeigte sich eine akzentuierte Reaktion der RSP sowie höhere Valenz- und Arousalratings in der Patientengruppe. Das subjektiv intensivere Empfinden der Stimuli bei den Patienten hing zusätzlich mit emotionaler und sozialer Kompetenz zusammen.
In dieser Dissertation konnte gezeigt werden, dass Ausdrücke intensiver Emotionen im Gesicht oft als zweideutig wahrgenommen werden. Um ein genaueres Verständnis der Emotionswahrnehmung bei depressiven Menschen zu erlangen, konnten zudem mehrere Parameter des ANS identifiziert werden, die teils noch nicht untersucht wurden und einer intensiveren Emotionswahrnehmung bei depressiven Patienten zugrunde liegen könnten.
Hierbei wurden zusätzlich Zusammenhänge zu weiteren Aspekten der Depression, wie Defiziten in sozialen Kompetenzen, aufgezeigt. Damit gibt diese Dissertation umfassende Aufschlüsse über Emotionsverarbeitungsprozesse bei gesunden und depressiven Menschen.
Ocular gene therapy approaches have been developed for a variety of different diseases. In particular, clinical gene therapy trials for RPE65 mutations, X-linked retinoschisis, and choroideremia have been conducted at different centers in recent years, showing that adeno-associated virus (AAV)-mediated gene therapy is safe, but limitations exist as to the therapeutic benefit and long-term duration of the treatment. The technique of vector delivery to retinal cells relies on subretinal injection of the vector solution, causing a transient retinal detachment. Although retinal detachments are known to cause remodeling of retinal neuronal structures as well as significant cell loss, the possible effects of this short-term therapeutic retinal detachment on retinal structure and circuitry have not yet been studied in detail. In this study, retinal morphology and apoptotic status were examined in healthy rat retinas following AAV-mediated gene transfer via subretinal injection with AAV2/5.CMV.d2GFP or sham injection with fluorescein. Outer plexiform layer (OPL) morphology was assessed by immunohistochemical labeling, laser scanning confocal microscopy, and electron microscopy. The number of synaptic contacts in the OPL was quantified after labeling with structural markers. To assess the apoptotic status, inflammatory and pro-apoptotic markers were tested and TUNEL assay for the detection of apoptotic nuclei was performed. Pre- and postsynaptic structures in the OPL, such as synaptic ribbons or horizontal and bipolar cell processes, did not differ in size or shape in injected versus non-injected areas and control retinas. Absolute numbers of synaptic ribbons were not altered. No signs of relevant gliosis were detected. TUNEL labeling of retinal cells did not vary between injected and non-injected areas, and apoptosis-inducing factor was not delocalized to the nucleus in transduced areas. The neuronal circuits in the OPL of healthy rat retinas undergoing AAV-mediated gene transfer were not altered by the temporary retinal detachment caused by subretinal injection, the presence of viral particles, or the expression of green fluorescent protein as a transgene. This observation likely requires further investigations in the dog model for RPE65 deficiency in order to determine the impact of RPE65 transgene expression on diseased retinas in animals and men.
Hyponatremia is the most common and by far underestimated electrolyte disorder in clinical practice. Especially in elderly patients, treatment of symptomatic hyponatremia is challenging. Herein we describe the case of an octogenarian with recurrent symptomatic hyponatremia due to idiopathic syndrome of inappropriate antidiuretic hormone release (SIADH). Fluid restriction was insufficient to prevent repeated episodes of hyponatremia complicated by falls and coma. After introduction of a low-dose therapy with tolvaptan, serum sodium levels as well as the clinical condition were stable under vaptan therapy, without any relapse for more than six years now. This case demonstrates that long-term tolvaptan treatment for hyponatremia caused by SIADH is safe and well tolerated, even in the elderly.