Investigation of EBT2 and EBT3 films for proton dosimetry in the 4-20 MeV energy range.

Radiochromic films such as Gafchromic EBT2 or EBT3 films are widely used for dose determination in radiation therapy because they offer a superior spatial resolution compared to any other digital dosimetric 2D detector array. The possibility to detect steep dose gradients is not only attractive for intensity-modulated radiation therapy with photons but also for intensity-modulated proton therapy. Their characteristic dose rate-independent response makes radiochromic films also attractive for dose determination in cell irradiation experiments using laser-driven ion accelerators, which are currently being investigated as future medical ion accelerators. However, when using these films in ion beams, the energy-dependent dose response in the vicinity of the Bragg peak has to be considered. In this work, the response of these films for low-energy protons is investigated. To allow for reproducible and background-free irradiation conditions, the films were exposed to mono-energetic protons from an electrostatic accelerator, in the 4-20 MeV energy range. For comparison, irradiation with clinical photons was also performed. It turned out that in general, EBT2 and EBT3 films show a comparable performance. For example, dose-response curves for photons and protons with energies as low as 11 MeV show almost no differences. However, corrections are required for proton energies below 11 MeV. Care has to be taken when correction factors are related to an average LET from depth-dose measurements, because only the dose-averaged LET yields similar results as obtained in mono-energetic measurements.

Link between supercurrent diode and anomalous Josephson effect revealed by gate-controlled interferometry.

In Josephson diodes the asymmetry between positive and negative current branch of the current-phase relation leads to a polarity-dependent critical current and Josephson inductance. The supercurrent nonreciprocity can be described as a consequence of the anomalous Josephson effect -a φ0-shift of the current-phase relation- in multichannel ballistic junctions with strong spin-orbit interaction. In this work, we simultaneously investigate φ0-shift and supercurrent diode efficiency on the same Josephson junction by means of a superconducting quantum interferometer. By electrostatic gating, we reveal a direct link between φ0-shift and diode effect. Our findings show that spin-orbit interaction in combination with a Zeeman field plays an important role in determining the magnetochiral anisotropy and the supercurrent diode effect.

Sign reversal of the Josephson inductance magnetochiral anisotropy and 0-π-like transitions in supercurrent diodes.

The recent discovery of the intrinsic supercurrent diode effect, and its prompt observation in a rich variety of systems, has shown that non-reciprocal supercurrents naturally emerge when both space-inversion and time-inversion symmetries are broken. In Josephson junctions, non-reciprocal supercurrent can be conveniently described in terms of spin-split Andreev states. Here we demonstrate a sign reversal of the Josephson inductance magnetochiral anisotropy, a manifestation of the supercurrent diode effect. The asymmetry of the Josephson inductance as a function of the supercurrent allows us to probe the current-phase relation near equilibrium, and to probe jumps in the junction ground state. Using a minimal theoretical model, we can then link the sign reversal of the inductance magnetochiral anisotropy to the so-called 0-π-like transition, a predicted but still elusive feature of multichannel junctions. Our results demonstrate the potential of inductance measurements as sensitive probes of the fundamental properties of unconventional Josephson junctions.

No proinflammatory signature in CD34+ hematopoietic progenitor cells in multiple sclerosis patients.

Autologous hematopoietic stem cell transplantation (aHSCT) has been used as a therapeutic approach in multiple sclerosis (MS). However, it is still unclear if the immune system that emerges from autologous CD34+ hematopoietic progenitor cells (HPC) of MS patients is pre-conditioned to re-develop the proinflammatory phenotype. The objective of this article is to compare the whole genome gene and microRNA expression signature in CD34+ HPC of MS patients and healthy donors (HD). CD34+ HPC were isolated from peripheral blood of eight MS patients and five HD and analyzed by whole genome gene expression and microRNA expression microarray. Among the differentially expressed genes (DEGs) only TNNT1 reached statistical significance (logFC=3.1, p<0.01). The microRNA expression was not significantly different between MS patients and HD. We did not find significant alterations of gene expression or microRNA profiles in CD34+ HPCs of MS patients. Our results support the use of aHSCT for treatment of MS.

Comparison of Gafchromic EBT2 and EBT3 films for clinical photon and proton beams.

PURPOSE: Dose verification in highly conformal radiation therapy such as IMRT or proton therapy can benefit from the high spatial resolution offered by radio-chromic films such as Gafchromic EBT or EBT2. Recently, a new generation of these films, EBT3, has become available. The composition and thickness of the sensitive layer are the same as for the previous EBT2 films. The most important change is the symmetric layer configuration to eliminate side orientation dependence, which is reported for EBT2 films. METHODS: The general film characteristics such as sensitivity to read-out orientation and postexposure darkening evolution of the new EBT3 film are evaluated. Film response has been investigated in clinical photon and proton beams and compared to former EBT2 films. Quenching effects in the proton Bragg peak region have been studied for both, EBT2 and EBT3 films. RESULTS: The general performance of EBT3 is comparable to EBT2, and the orientation dependence with respect to film side is completely eliminated in EBT3 films. Response differences of EBT2 and EBT3 films are of the same order of magnitude as batch-to-batch variations observed for EBT2 films. No significant difference has been found for both generations of EBT films between photon and proton exposure. Depth dose measurements of EBT2 and EBT3 show an excellent agreement, though underestimating dose by up to 20% in the Bragg peak region. CONCLUSIONS: The symmetric configuration of EBT3 presents a major improvement for film handling. EBT3 has similar dosimetric performance as its precursor EBT2 and can, thus, be applied to dose verification in IMRT in the same way. For dose verification in proton therapy the underresponse in the Bragg peak region has to be taken into account.

Effect of Rashba and Dresselhaus spin-orbit coupling on supercurrent rectification and magnetochiral anisotropy of ballistic Josephson junctions.

Simultaneous breaking of inversion- and time-reversal symmetry in Josephson junction (JJ) leads to a possible violation of theI(φ) = -I(-φ) equality for the current-phase relation. This is known as anomalous Josephson effect and it produces a phase shiftφ0in sinusoidal current-phase relations. In ballistic JJs with non-sinusoidal current phase relation the observed phenomenology is much richer, including the supercurrent diode effect and the magnetochiral anisotropy (MCA) of Josephson inductance. In this work, we present measurements of both effects on arrays of JJs defined on epitaxial Al/InAs heterostructures. We show that the orientation of the current with respect to the lattice affects the MCA, possibly as the result of a finite Dresselhaus component. In addition, we show that the two-fold symmetry of the Josephson inductance reflects in the activation energy for phase slips.

A deep-UV optical frequency comb at 205 nm.

By frequency quadrupling a picosecond pulse train from a Ti:sapphire laser at 820 nm we generate a frequency comb at 205 nm with nearly bandwidth-limited pulses. The nonlinear frequency conversion is accomplished by two successive frequency doubling stages that take place in resonant cavities that are matched to the pulse repetition rate of 82 MHz. This allows for an overall efficiency of 4.5 % and produces an output power of up to 70 mW for a few minutes and 25 mW with continuous operation for hours. Such a deep UV frequency comb may be employed for direct frequency comb spectroscopy in cases where it is less efficient to convert to these short wavelengths with continuous wave lasers.

Selective adhesion of cells from different telencephalic regions.

We asked whether specifications of different regions of the rodent and avian telencephalon during development involved the acquisition of differential adhesive properties. Cells from different regions were aggregated in a short-term aggregation assay, and their segregation was analyzed. Both neurons and precursor cells from cortex segregate from striatal cells at early, but not later, stages, whereas cells from rodent neocortex and hippocampus segregated only during later stages. Segregation was abolished when Ca2+-dependent but not Ca2+-independent adhesion molecules were selectively removed. Thus, selective adhesion appears to be a conserved mechanism that restricts cellular mixing and might serve to maintain positional information during forebrain development. A candidate for mediating the Ca2+-dependent segregation is the CD15 (Lewis(x)) carbohydrate epitope, which is selectively expressed by mammalian cortex but not striatum.

The anatomy of the stigma and style from Cyclamen persicum (Mill.) cv. "pure white" and its relation to pollination success.

Old flowers of Cyclamen generate few or no seeds. To understand the pollination problems of Cyclamen we investigate the general anatomy of the stigma and the style of Cyclamen persicum by scanning electron microscopy at different stages of floral maturity. Our investigations confirm that there is a hollow style. Against data commonly found in the literature, we present evidence of pollen germination and tube growth that show the stigma is not outside the style but inside it. Furthermore the maturation process of the style during the flowering time indicates a mechanism by which the stigma becomes shut off through closure at the terminal aperture of the style. At 3 to 5 days after anthesis there was the beginning closure of the style which was nearly completed at 21 days. The substance which leads to the closure is still unknown. The closure of the hollow style is a probable cause for failure of seed set in flowers not pollinated early in anthesis.

Severity-dependent differences in early management of thoracic trauma in severely injured patients - Analysis based on the TraumaRegister DGU®.

BACKGROUND: Major trauma is associated with chest injuries in nearly 50% of multiple injuries. Thoracic trauma is a relevant source of comorbidity throughout the period of multiply-injured patient care and may require swift and well-thought-out interventions in order to avert a deleterious outcome. In this epidemiological study we seek to characterize groups of different thoracic trauma severity in severely injured patients and identify related differences in prehospital and early clinical management. This may help to anticipate necessary treatment for chest injuries. METHODS: Patients documented between 2002 and 2012 in the TraumaRegister DGU®, aged ≥ 16 years, determined Injury Severity Score ≥ 16, and documentation from European trauma centers were analyzed. Isolated brain injury and severe head injury (Abbreviated Injury ScaleHead ≥ 4) led to patient exclusion. Patient subgroups were formed according to the Abbreviated Injury ScaleThorax as Controls, AIS-2, AIS-3, AIS-4, and AIS-5/6. Demographic and clinical characteristics comparing the aforementioned groups were evaluated using descriptive statistics. RESULTS: Twenty two thousand five hundred sixty five predominantly male (74%) patients, mean age 45.7 years (SD 19.3), suffering from blunt trauma (95%), and presenting a mean Injury Severity Score of 25.6 (SD 9.6) were analyzed. Higher thoracic injury severity was associated with more different thoracic injuries. The highest rate of prehospital intubation (58%) occurred in AISThorax-5/6. The worse the chest trauma, the more chest tubes were placed prehospitally, peaking at 22% in AISThorax-5/6. Out-of-hospital cardiopulmonary resuscitation was successfully performed in 11% in AISThorax-5/6 compared to 1%-3% in lesser thoracic trauma severity. Massive transfusion and emergency surgery was highest in AISThorax-5/6 compared to lesser thoracic injury (12% vs. 5%-7% and 17% vs. 3%-7%) and both were independently associated with thoracic injuries in patients with AISThorax ≥ 4. CONCLUSIONS: We provide epidemiological data on trauma mechanism, concomitant injuries, frequencies of emergency interventions and outcome associated with different thoracic trauma severity. Prehospital and early clinical management is more complex when AISThorax is ≥ 4. Severely injured patients with critical thoracic trauma are most challenging to take care of with highest rates in prehospital intubation, cardiopulmonary resuscitation, chest tube placements, blood transfusions as well as emergency surgery.

Co-expression of the voltage-gated potassium channel Kv1.4 with transient receptor potential channels (TRPV1 and TRPV2) and the cannabinoid receptor CB1 in rat dorsal root ganglion neurons.

Potassium channels contribute to basic neuronal excitability and modulation. Here, we examined expression patterns of the voltage-gated potassium channel Kv1.4, the nociceptive transduction channels TRPV1 and TRPV2 as well as the putative anti-nociceptive cannabinoid receptor CB1 by immunofluorescence double-labelings in sections of rat dorsal root ganglia (DRGs). Kv1.4, TRPV1 and CB1 were each detected in about one third of neurons (35.7+/-0.5%, 29.4+/-1.1% and 36.4+/-0.5%, respectively, mean diameter 19.1+/-0.3 microm). TRPV2 was present in 4.4+/-0.4% of all neurons that were significantly larger in diameter (27.4+/-0.7 microm; P < 0.001). Antibody double-labeling revealed that the majority of Kv1.4-positive neurons co-expressed TRPV1 (73.9+/-1.5%) whereas none expressed TRPV2. The largest overlap was found with CB1 (93.1+/-0.1%). CB1 expression resembled that seen for Kv1.4 since the majority of neurons expressing CB1-protein also expressed TRPV1 (69.4+/-6.5%) but not TRPV2 (0.6+/-0.3%). When CB1-mRNA was detected using in situ hybridizations an additional subset of larger neurons was labeled including 82.4+/-17.7% of the TRPV2 expressing neurons. However, co-localization of Kv1.4 with CB1-mRNA (92%, mean diameter: 18.5 microm) was essentially the same as with CB1-protein. The almost complete overlap of CB1 and Kv1.4 in nociceptive DRG neurons suggests a functional synergistic action between Kv1.4 and CB1. The potassium channel may have two important roles in nociception. As the molecular basis of A-type current it could be involved in the control of repetitive discharges at peripheral terminals and as a downstream signal transduction site of CB1 in the control of presynaptic transmitter release at central terminals.

Effects of item sequence on the performance of the AUDIT in general practices.

BACKGROUND: One important task in identifying subjects with alcohol use disorders (AUDs) in the general medical practice setting is the development of effective screening instruments. Sensitivity of screening questionnaires might differ according to the introductory items. This study compares two versions of the alcohol use disorders identification test (AUDIT) with varied item sequence randomly applied to patients derived from a sample of general practitioners (GP) patients. METHODS: Participants were recruited from general practices in two northern German cities; they received two different versions of the AUDIT, one group receiving the original version starting with three items addressing frequency and quantity of alcohol use (AUDIT1), and a second group receiving a version in which these items were put at the end of the questionnaire (AUDIT2). In total, 10.803 screenings were conducted (refusal rate: 5%). Alcohol use disorders were diagnosed using the Munich-Composite International Diagnostic Interview (M-CIDI). RESULTS: Logistic regression analysis revealed that AUDIT1 subjects had higher scores in the consumption items of the AUDIT, whereas AUDIT2 subjects scored higher on items focussing on symptoms of alcohol dependence or abuse. CONCLUSION: The sequence upon which items of the AUDIT are presented influences the report of drinking patterns and symptoms of alcohol use disorders in GP patients.

Assessment of cytological criteria for diagnosing osteosarcoma in dogs.

OBJECTIVES: To evaluate the specific cytological criteria of osteosarcomas in dogs. METHODS: Significant cytological characteristics of osteosarcoma and benign mesenchymal bone proliferations were determined from imprint smears of 25 dogs with osteosarcoma (group 1) and 20 dogs admitted for removal of surgical bone implants after uncomplicated healing of bone fractures (group 2). RESULTS: Mild to moderate cellular necrosis occurred frequently in patients with osteosarcoma. The cytoplasm of osteoblasts was pale blue to blue with a more pronounced basophilia in group 2. In 48 per cent of the patients in group 1, but none in group 2, osteoblasts showed a slight to moderate eosinophilic cytoplasmic granulation. In both groups, osteoblasts contained one red to pale blue nucleus with one or two grey-red to blue nucleoli in group 2. Forty-four per cent of animals in group 1 had osteoblasts with more than two nucleoli per nucleus. The median nuclear:cytoplasmic ratio was higher in group 1 (1:2.0) than in group 2 (1:3.5). Osteoblasts in group I were frequently seen to have a clumped chromatin pattern and showed significantly more criteria of malignancy (median six criteria per patient) than those in group 2 (median two criteria per patient). In group 1, mitoses of osteoblasts were detectable in 23 of 25 dogs, whereas only one dog in group 2 had evidence of mitotic osteoblasts. CLINICAL SIGNIFICANCE: Cytological criteria can be helpful in the diagnosis of canine osteosarcoma.

[Implantable cardioverter-defibrillator at the end of life]. / Implantierbarer Kardioverter-Defibrillator am Ende des Lebens.

Brady- and tachyarrhythmias at the end of life are common observations. Implantable cardioverter-defibrillators answer with antibrady and antitachycardia pacing, which will not be associated with any complaints of the dying patient. In contrast, defibrillation and cardioversion shocks are extremely painful. Therefore shocks should be inactivated at the end of life. Family doctors, internists, emergency physicians and paramedics are unable to inactivate shocks. Deactivation of shocks at the end of life is not comparable to euthanasia or assisted suicide, but allow the patient to die at the end of an uncurable endstage disease. Deactivation of shocks should be discussed with the patient before initial implantation of the devices. The precise moment of the inactivation at the end of life should be discussed with patients and relatives. There is no common recommendation for the time schedule of this decision; therefore it should be based on the individual situation of the patient. Emergency health care physicians need magnets and sufficient information to inactivate defibrillators. The wishes of the patient have priority in the decision process and should be written in the patient's advance directive, which must be available in the final situation. However the physician must not necessarily follow every wish of the patient. As long as the laws in the European Union are not uniform, German recommendations are needed.

Ionoacoustic characterization of the proton Bragg peak with submillimeter accuracy.

PURPOSE: Range verification in ion beam therapy relies to date on nuclear imaging techniques which require complex and costly detector systems. A different approach is the detection of thermoacoustic signals that are generated due to localized energy loss of ion beams in tissue (ionoacoustics). Aim of this work was to study experimentally the achievable position resolution of ionoacoustics under idealized conditions using high frequency ultrasonic transducers and a specifically selected probing beam. METHODS: A water phantom was irradiated by a pulsed 20 MeV proton beam with varying pulse intensity and length. The acoustic signal of single proton pulses was measured by different PZT-based ultrasound detectors (3.5 and 10 MHz central frequencies). The proton dose distribution in water was calculated by Geant4 and used as input for simulation of the generated acoustic wave by the matlab toolbox k-WAVE. RESULTS: In measurements from this study, a clear signal of the Bragg peak was observed for an energy deposition as low as 10(12) eV. The signal amplitude showed a linear increase with particle number per pulse and thus, dose. Bragg peak position measurements were reproducible within ±30 µm and agreed with Geant4 simulations to better than 100 µm. The ionoacoustic signal pattern allowed for a detailed analysis of the Bragg peak and could be well reproduced by k-WAVE simulations. CONCLUSIONS: The authors have studied the ionoacoustic signal of the Bragg peak in experiments using a 20 MeV proton beam with its correspondingly localized energy deposition, demonstrating submillimeter position resolution and providing a deep insight in the correlation between the acoustic signal and Bragg peak shape. These results, together with earlier experiments and new simulations (including the results in this study) at higher energies, suggest ionoacoustics as a technique for range verification in particle therapy at locations, where the tumor can be localized by ultrasound imaging. This acoustic range verification approach could offer the possibility of combining anatomical ultrasound and Bragg peak imaging, but further studies are required for translation of these findings to clinical application.

Analysis of NO synthase expression in neuronal, astroglial and fibroblast-like derivatives differentiating from PCC7-Mz1 embryonic carcinoma cells.

We studied the expression of the NO synthase isoforms in an in vitro model of neural development using RT-PCR, Western blot and immunohistochemistry. Murine PCC7-Mz1 cells (Jostock et al., Eur. J. Cell Biol. 76, 63-76, 1998) differentiate in the presence of all-trans retinoic acid and dibutyryl cAMP along the neural pathway into neuron-like, fibroblast-like and astroglia-like cells. Undifferentiated cells showed immunofluorescent staining for neuronal-type NOS I and endothelial-type NOS III. This expression pattern was retained in those cells differentiating into neurofilament- and tau protein-positive neuronal cells. Thymocyte alloantigen (Thy1.2/CD 90.2)-positive fibroblasts, appearing around day 3, and glial fibrillary acidic protein (GFAP)-positive astroglial cells, appearing after day 6 of differentiation, stained negative for any NOS isoform. Starting at day 6 of differentiation, expression of inducible-type NOS II could be stimulated with cytokines in a subset of cells, which may represent activated astrocytes. NOS II was always undetectable in non-induced cultures. These data indicate that the ability of stem cells to express NOS I and NOS III is only retained when the cells differentiate along the neuronal lineage, while a small subpopulation of cells acquires the ability to express NOS II in response to cytokines.

De novo acquisition of neuronal polarity in retinoic acid-induced embryonal carcinoma cells.

The mouse embryonal carcinoma cell line PCC7-Mz1 represents an advantageous model to study acquisition of polarity by neurons. During the first two days after differentiation is induced by the addition of retinoic acid, the neuronal derivatives develop extensions which for at least four more days do not differ from each other in growth characteristics, morphology, and marker expression. Beginning around differentiation day 6 and following the relocation of the nucleus from a central to a polar position in the cell soma, the morphology and marker expression changes dramatically: expression of MAP2 diminishes and eventually disappears in the thinner neurite (future axon), which originates at the nucleated pole, but remains strong in the branched, broad based neurite(s). The opposite changes in expression are observed for synaptophysin, together with a clustering of the vesicle protein in varicosity-like areas. Complete segregation of expression of the two markers is achieved around day 12, shortly followed by dendrite-specific location of MAP2 mRNA and the ability to generate and conduct action potentials. Our studies add several aspects to the process of neuronal polarity acquisition, as it was previously studied in primary cultures of embryonic neurons: (i) we monitored neuronal differentiation from the birth of neurons, rather than from later and less defined maturation stages, (ii) cell nucleus relocation may be associated with the induction of neuronal polarity, and (iii) functional competence of neurons is closely associated with previous acquisition of polarity. Acquisition of polarity by PCC7-Mz1 neuronal derivatives probably refers to de novo acquisition rather than to reestablishment of polarity.

Functional and structural analysis of acetylcholine receptor-rich membranes after negative staining.

Phosphotungstate (pH 7.4) used for negative staining of membranes from Torpedo electric tissue rich in acetylcholine receptor does not affect binding properties and cation permeability of the receptor and its ion channel. Uranyl salts, frequently used for negative staining, precipitate the receptor-rich membranes making measurements of ligand binding and ion-permeability regulation impossible. The gross ultrastructure in the two stains is not significantly different, but for future high-resolution electron microscopy aiming at visualizing structural details of functional receptor molecules it is necessary to resort to a stain preserving native and active receptor. Uranyl salts are not applicable for this purpose. The electron micrographs obtained with phosphotungstate reveal two distinct structures in the receptor-rich membrane: a closed ring ('doughnut') and an open ring ('horseshoe'), with a ratio of abundance of about 3:2.

The murine nuclear orphan receptor GCNF is expressed in the XY body of primary spermatocytes.

We have studied the expression of the nuclear orphan receptor GCNF (germ cell nuclear factor) on the mRNA and protein level in pubertal and adult mouse testes. We show by Northern and Western blot analyses and by in situ hybridization that GCNF is expressed in spermatocytes and round spermatids of adult mouse testis suggesting that GCNF may be a transcriptional regulator of spermatogenesis. Since the GCNF protein is accumulated in the XY body of late pachytene spermatocytes, it may be involved in transcriptional inactivation of sex chromosomes.

Modulation of voltage-gated K(+) channels Kv11 and Kv1 4 by forskolin.

Forskolin (FSK) affects voltage-gated K(+) (Kv) currents in different cell types, but it is not known which of the various subunits form FSK-sensitive Kv channels. We compared the effect of the compound at Kv1.1 and Kv1.4 channels ectopically expressed in HEK 293 cells. Low FSK concentrations induced a phosphorylation-dependent potentiation of Kv1.1 currents. At higher concentrations, this effect was superimposed by a fast, cAMP-independent channel block. Kv1.4 currents were inhibited with lower potency by FSK but were not modified by phosphorylation. The variable effect of the compound might help to distinguish between Kv subunits expressed by native cells.