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Institute
- Theodor-Boveri-Institut für Biowissenschaften (100)
- Medizinische Klinik und Poliklinik II (37)
- Physikalisches Institut (34)
- Institut für Psychologie (33)
- Neurologische Klinik und Poliklinik (32)
- Graduate School of Life Sciences (29)
- Medizinische Klinik und Poliklinik I (25)
- Julius-von-Sachs-Institut für Biowissenschaften (23)
- Rudolf-Virchow-Zentrum (23)
- Institut für Theoretische Physik und Astrophysik (21)
Schriftenreihe
Sonstige beteiligte Institutionen
- Bavarian Center for Applied Energy Research e.V. (ZAE Bayern) (1)
- Center for Nanosystems Chemistry (1)
- Department of Chemistry, Sungkyunkwan University, 440-746 Suwon, Republic of Korea (1)
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"...using different names, as Zeus and Dis" (Arist 16). Concepts of "God" in the letter of Aristeas
(2016)
The “Letter of Aristeas” recounts the translations of the Hebrew Bible into Greek. Probably originating in the 2nd century BCE1, the book tells a legend of how the translation of the Torah into Greek came into being. This shows that translating a holy, canonical text or the first time needed explication. Notably, the translation of the godly nomos (Arist 3) comparatively takes up little space (Arist 301–307). And it has to be noted, that “God” is seldom a topic in the Book of Aristeas. The word (ὁ) θεός “God” is found in only three contexts: in the dialogue between king Ptolemaios and Aristeas (Arist 15–21), in the dialogue of the high priest Eleazar and Aristeas (Arist 121–171; above all 128; 130–141; 155–166; 168) and in the question-and-answer-speech during the symposium at the Ptolemaic royal court between the king and the Jewish scholars (Arist 184–294).
In analysing the different statements regarding God, the frame of the narrative is of decisive importance: In the Book of Aristeas, “Aristeas” (Ἀριστέας), who writes in Greek, presents himself as the author, but he is also part of the story. Accordingly, Aristeas is the narrator, who tells the story from his own point of view, and at the same time, he is a character in the ‘world’ of the text. This Aristeas presents himself as a Greek and a Non-Jew (Arist 16; 121–171), who already wrote a book (Arist 6) and plans further publications (Arist 322). In the double-role as narrator of the text and protagonist in the text, Aristeas has to be differentiated from the (real) writer/author of the Book of Aristeas, who possibly was Jewish. That means that the (real, probably Jewish) author of the Book of Aristeas presents (or invents) “Aristeas” and gives him the role of the narrator of his text.3 The author portrays Aristeas as a Greek, non-Jewish character, who is a servant of the royal court. This differentiation between narrator and writer/author is of crucial importance for the question of the different conceptions of God in the Book of Aristeas.
Although Lijphart's typology of consensus and majoritarian democracy can be regarded as the most widely used tool to classify democratic regimes, it has been rarely applied to Latin America so far. We try to fill this gap by adapting Lijphart's typological framework to the Latin American context in the following way. In contrast to previous studies, we treat the type of democracy as an independent variable and include informal factors such as clientelism or informal employment in our assessment of democratic patterns. On this basis, we aim to answer the following questions. First, how did the patterns of democracy evolve in Latin America over the two decades between 1990 and 2010 and what kind of differences can be observed in the region? Second, what are the institutional determinants of the observed changes? We focus on the emergence of new parties because of their strong impact on the first dimension of Lijphart's typology. From our observations we draw the following tentative conclusions: If strong new parties established themselves in the party system but failed to gain the presidency, they pushed the system towards consensualism. Conversely, new parties that gained the presidency produced more majoritarian traits.
Background
Mobile 3D fluoroscopes have become increasingly available in neurosurgical operating rooms. In this series, the image quality and value of intraoperative 3D fluoroscopy with intravenous contrast agent for the evaluation of aneurysm occlusion and vessel patency after clip placement was assessed in patients who underwent surgery for intracranial aneurysms.
Materials and methods
Twelve patients were included in this retrospective analysis. Prior to surgery, a 360° rotational fluoroscopy scan was performed without contrast agent followed by another scan with 50 ml of intravenous iodine contrast agent. The image files of both scans were transferred to an Apple PowerMac® workstation, subtracted and reconstructed using OsiriX® free software. The procedure was repeated after clip placement. Both image sets were compared for assessment of aneurysm occlusion and vessel patency.
Results
Image acquisition and contrast administration caused no adverse effects. Image quality was sufficient to follow the patency of the vessels distal to the clip. Metal artifacts reduce the assessability of the immediate vicinity of the clip. Precise image subtraction and post-processing can reduce metal artifacts and make the clip-site assessable and depict larger neck-remnants.
Conclusion
This technique quickly supplies images at adequate quality to evaluate distal vessel patency after aneurysm clipping. Significant aneurysm remnants may be depicted as well. As it does not require visual control of all vessels that are supposed to be evaluated intraoperatively, this technique may be complementary to other intraoperative tools like indocyanine green videoangiography and micro-Doppler, especially for the assessment of larger aneurysms. At the momentary state of this technology, it cannot replace postoperative conventional angiography. However, 3D fluoroscopy and image post-processing are young technologies. Further technical developments are likely to result in improved image quality.
The Josephson effect describes the generic appearance of a supercurrent in a weak link between two superconductors. Its exact physical nature deeply influences the properties of the supercurrent. In recent years, considerable efforts have focused on the coupling of superconductors to the surface states of a three-dimensional topological insulator. In such a material, an unconventional induced p-wave superconductivity should occur, with a doublet of topologically protected gapless Andreev bound states, whose energies vary 4π-periodically with the superconducting phase difference across the junction. In this article, we report the observation of an anomalous response to rf irradiation in a Josephson junction made of a HgTe weak link. The response is understood as due to a 4π-periodic contribution to the supercurrent, and its amplitude is compatible with the expected contribution of a gapless Andreev doublet. Our work opens the way to more elaborate experiments to investigate the induced superconductivity in a three-dimensional insulator.
Two 5-methylcytosine (5-MeC)-rich heterochromatic regions were demonstrated in metaphase chromosomes of the Indian muntjac by indirect immunofluorescence using a monoclonal anti-5-MeC antibody. The metaphases were obtained from diploid and triploid cell lines. A major region is located in the ‘neck' of the 3;X fusion chromosome and can be detected after denaturation of the chromosomal DNA with UV-light irradiation for 1 h. It is located exactly at the border of the X chromosome and the translocated autosome 3. A minor region is found in the centromeric region of the free autosome 3 after denaturing the chromosomal DNA for 3 h or longer. The structure and possible function of the major hypermethylated region as barrier against spreading of the X-inactivation process into the autosome 3 is discussed.
Background
\(^{177}\)Lu is used in peptide receptor radionuclide therapies for the treatment of neuroendocrine tumors. Based on the recent literature, SST2 antagonists are superior to agonists in tumor uptake. The compound OPS201 is the novel somatostatin antagonist showing the highest SST2 affinity. The aim of this study was to measure the in vivo biodistribution and dosimetry of \(^{177}\)Lu-OPS201 in five anesthetized Danish Landrace pigs as an appropriate substitute for humans to quantitatively assess the absorbed doses for future clinical applications.
Results
\(^{177}\)Lu-OPS201 was obtained with a specific activity ranging from 10 to 17 MBq/μg. Prior to administration, the radiochemical purity was measured as s > 99.7 % in all cases. After injection, fast clearance of the compound from the blood stream was observed. Less than 5 % of the injected activity was presented in blood 10 min after injection. A series of SPECT/CT and whole-body scans conducted until 10 days after intravenous injection showed uptake mostly in the liver, spine, and kidneys. There was no visible uptake in the spleen. Blood samples were taken to determine the time-activity curve in the blood. Time-activity curves and time-integrated activity coefficients were calculated for the organs showing visible uptake. Based on these data, the absorbed organ dose coefficients for a 70-kg patient were calculated with OLINDA/EXM. For humans after an injection of 5 GBq \(^{177}\)Lu-OPS201, the highest predicted absorbed doses are obtained for the kidneys (13.7 Gy), the osteogenic cells (3.9 Gy), the urinary bladder wall (1.8 Gy), and the liver (1.0 Gy). No metabolites of 177Lu-OPS201 were found by radio HPLC analysis. None of the absorbed doses calculated will exceed organ toxicity levels.
Conclusions
The \(^{177}\)Lu-OPS201 was well tolerated and caused no abnormal physiological or behavioral signs. In vivo distributions and absorbed doses of pigs are comparable to those observed in other publications. According to the biodistribution data in pigs, presented in this work, the expected radiation exposure in humans will be within the acceptable range.
Chemokine receptor-4 (CXCR4) has been reported to be overexpressed in glioblastoma (GBM) and to be associated with poor survival. This study investigated the feasibility of non-invasive CXCR4-directed imaging with positron emission tomography/computed tomography (PET/CT) using the radiolabelled chemokine receptor ligand \(^{68}\)Ga-Pentixafor.
15 patients with clinical suspicion on primary or recurrent glioblastoma (13 primary, 2 recurrent tumors) underwent \(^{68}\)Ga-Pentixafor-PET/CT for assessment of CXCR4 expression prior to surgery. O-(2-\(^{18}\)F-fluoroethyl)-L-tyrosine (\(^{18}\)F-FET) PET/CT images were available in 11/15 cases and were compared visually and semi-quantitatively (SUV\(_{max}\), SUV\(_{mean}\)). Tumor-to-background ratios (TBR) were calculated for both PET probes. \(^{68}\)Ga-Pentixafor-PET/CT results were also compared to histological CXCR4 expression on neuronavigated surgical samples.
\(^{68}\)Ga-Pentixafor-PET/CT was visually positive in 13/15 cases with SUV\(_{mean}\) and SUV\(_{max}\) of 3.0±1.5 and 3.9±2.0 respectively. Respective values for \(^{18}\)F-FET were 4.4±2.0 (SUV\(_{mean}\)) and 5.3±2.3 (SUV\(_{max}\)). TBR for SUV\(_{mean}\) and SUV\(_{max}\) were higher for \(^{68}\)Ga-Pentixafor than for \(^{18}\)F-FET (SUV\(_{mean}\) 154.0±90.7 vs. 4.1±1.3; SUV\(_{max}\) 70.3±44.0 and 3.8±1.2, p<0.01), respectively. Histological analysis confirmed CXCR4 expression in tumor areas with high \(^{68}\)Ga-Pentixafor uptake; regions of the same tumor without apparent \(^{68}\)Ga-Pentixafor uptake showed no or low receptor expression.
In this pilot study, \(^{68}\)Ga-Pentixafor retention has been observed in the vast majority of glioblastoma lesions and served as readout for non-invasive determination of CXCR4 expression. Given the paramount importance of the CXCR4/SDF-1 axis in tumor biology, \(^{68}\)Ga-Pentixafor-PET/CT might prove a useful tool for sensitive, non-invasive in-vivo quantification of CXCR4 as well as selection of patients who might benefit from CXCR4-directed therapy.
To compare the effects of a 3‐week multimodal rehabilitation involving supervised high‐intensity interval training (HIIT) on female breast cancer survivors with respect to key variables of aerobic fitness, body composition, energy expenditure, cancer‐related fatigue, and quality of life to those of a standard multimodal rehabilitation program. A randomized controlled trial design was administered. Twenty‐eight women, who had been treated for cancer were randomly assigned to either a group performing exercise of low‐to‐moderate intensity (LMIE; n = 14) or a group performing high‐intensity interval training (HIIT; n = 14) as part of a 3‐week multimodal rehabilitation program. No adverse events related to the exercise were reported. Work economy improved following both HIIT and LMIE, with improved peak oxygen uptake following LMIE. HIIT reduced mean total body fat mass with no change in body mass, muscle or fat‐free mass (best P < 0.06). LMIE increased muscle and total fat‐free body mass. Total energy expenditure (P = 0.45) did not change between the groups, whereas both improved quality of life to a similar high extent and lessened cancer‐related fatigue. This randomized controlled study demonstrates that HIIT can be performed by female cancer survivors without adverse health effects. Here, HIIT and LMIE both improved work economy, quality of life and cancer‐related fatigue, body composition or energy expenditure. Since the outcomes were similar, but HIIT takes less time, this may be a time‐efficient strategy for improving certain aspects of the health of female cancer survivors.
We present a supersymmetric left-right model which predicts gauge coupling unification close to the string scale and extra vector bosons at the TeV scale. The subtleties in constructing a model which is in agreement with the measured quark masses and mixing for such a low left-right breaking scale are discussed. It is shown that in the constrained version of this model radiative breaking of the gauge symmetries is possible and a SM-like Higgs is obtained. Additional CP-even scalars of a similar mass or even much lighter are possible. The expected mass hierarchies for the supersymmetric states differ clearly from those of the constrained MSSM. In particular, the lightest down-type squark, which is a mixture of the sbottom and extra vector-like states, is always lighter than the stop. We also comment on the model’s capability to explain current anomalies observed at the LHC.
Purpose: A study of real-time adaptive radiotherapy systems was performed to test the hypothesis that, across delivery systems and institutions, the dosimetric accuracy is improved with adaptive treatments over non-adaptive radiotherapy in the presence of patient-measured tumor motion. Methods and materials: Ten institutions with robotic(2), gimbaled(2), MLC(4) or couch tracking(2) used common materials including CT and structure sets, motion traces and planning protocols to create a lung and a prostate plan. For each motion trace, the plan was delivered twice to a moving dosimeter; with and without real-time adaptation. Each measurement was compared to a static measurement and the percentage of failed points for gamma-tests recorded. Results: For all lung traces all measurement sets show improved dose accuracy with a mean 2%/2 mm gamma-fail rate of 1.6% with adaptation and 15.2% without adaptation (p < 0.001). For all prostate the mean 2%/2 mm gamma-fail rate was 1.4% with adaptation and 17.3% without adaptation (p < 0.001). The difference between the four systems was small with an average 2%/2 mm gamma-fail rate of <3% for all systems with adaptation for lung and prostate. Conclusions: The investigated systems all accounted for realistic tumor motion accurately and performed to a similar high standard, with real-time adaptation significantly outperforming non-adaptive delivery methods.
The Venus flytrap, \textit{Dionaea muscipula}, with its carnivorous life-style and its highly
specialized snap-traps has fascinated biologist since the days of Charles Darwin. The
goal of the \textit{D. muscipula} genome project is to gain comprehensive insights into the
genomic landscape of this remarkable plant.
The genome of the diploid Venus flytrap with an estimated size between 2.6 Gbp to
3.0 Gbp is comparatively large and comprises more than 70 % of repetitive regions.
Sequencing and assembly of genomes of this scale are even with state-of-the-art
technology and software challenging. Initial sequencing and assembly of the genome
was performed by the BGI (Beijing Genomics Institute) in 2011 resulting in a 3.7 Gbp
draft assembly. I started my work with thorough assessment of the delivered assembly
and data. My analysis showed that the BGI assembly is highly fragmented and
at the same time artificially inflated due to overassembly of repetitive sequences.
Furthermore, it only comprises about on third of the expected genes in full-length,
rendering it inadequate for downstream analysis.
In the following I sought to optimize the sequencing and assembly strategy to obtain
an assembly of higher completeness and contiguity by improving data quality and
assembly procedure and by developing tailored bioinformatics tools. Issues with
technical biases and high levels of heterogeneity in the original data set were solved
by sequencing additional short read libraries from high quality non-polymorphic DNA
samples. To address contiguity and heterozygosity I examined numerous alternative
assembly software packages and strategies and eventually identified ALLPATHS-LG
as the most suited program for assembling the data at hand. Moreover, by utilizing
digital normalization to reduce repetitive reads, I was able to substantially reduce
computational demands while at the same time significantly increasing contiguity of
the assembly.
To improve repeat resolution and scaffolding, I started to explore the novel PacBio
long read sequencing technology. Raw PacBio reads exhibit high error rates of 15 %
impeding their use for assembly. To overcome this issue, I developed the PacBio
hybrid correction pipeline proovread (Hackl et al., 2014). proovread uses high
coverage Illumina read data in an iterative mapping-based consensus procedure to
identify and remove errors present in raw PacBio reads. In terms of sensitivity and
accuracy, proovread outperforms existing software. In contrast to other correction
programs, which are incapable of handling data sets of the size of D. muscipula
project, proovread’s flexible design allows for the efficient distribution of work load on high-performance computing clusters, thus enabling the correction of the Venus
flytrap PacBio data set.
Next to the assembly process itself, also the assessment of the large de novo draft
assemblies, particularly with respect to coverage by available sequencing data, is
difficult. While typical evaluation procedures rely on computationally extensive
mapping approaches, I developed and implemented a set of tools that utilize k-mer
coverage and derived values to efficiently compute coverage landscapes of large-scale
assemblies and in addition allow for automated visualization of the of the obtained
information in comprehensive plots.
Using the developed tools to analyze preliminary assemblies and by combining my
findings regarding optimizations of the assembly process, I was ultimately able to
generate a high quality draft assembly for D. muscipula. I further refined the assembly
by removal of redundant contigs resulting from separate assembly of heterozygous
regions and additional scaffolding and gapclosing using corrected PacBio data. The
final draft assembly comprises 86 × 10 3 scaffolds and has a total size of 1.45 Gbp.
The difference to the estimated genomes size is well explained by collapsed repeats.
At the same time, the assembly exhibits high fractions full-length gene models,
corroborating the interpretation that the obtained draft assembly provides a complete
and comprehensive reference for further exploration of the fascinating biology of the
Venus flytrap.
Tissue-engineered skin equivalents mimic key aspects of the human skin, and can thus be employed as wound coverage for large skin defects or as in vitro test systems as an alternative to animal models. However, current skin equivalents lack a functional vasculature limiting clinical and research applications. This study demonstrates the generation of a vascularized skin equivalent with a perfused vascular network by combining a biological vascularized scaffold (BioVaSc) based on a decellularized segment of a porcine jejunum and a tailored bioreactor system. Briefly, the BioVaSc was seeded with human fibroblasts, keratinocytes, and human microvascular endothelial cells. After 14 days at the air-liquid interface, hematoxylin & eosin and immunohistological staining revealed a specific histological architecture representative of the human dermis and epidermis including a papillary-like architecture at the dermal-epidermal-junction. The formation of the skin barrier was measured non-destructively using impedance spectroscopy. Additionally, endothelial cells lined the walls of the formed vessels that could be perfused with a physiological volume flow. Due to the presence of a complex in-vivo-like vasculature, the here shown skin equivalent has the potential for skin grafting and represents a sophisticated in vitro model for dermatological research.
„Perspective-taking“ is the ability to put yourself into the place of somebody else. Psychological research distinguishes three kinds of perspective-taking, namely, perceptual (visuo-spatial), affective (empathy), and cognitive (theory of mind) perspective-taking. The last two kinds of perspective-taking are often summarized as “psychological perspective-taking”. This dissertation tackles the question of whether these three kinds of perspective-taking should be conceptualized as independent constructs or as facets of one and the same construct.
Prior research findings concerning this are equivocal. While some authors consider correlations between the different kinds of perspective-taking as too low for a unitary construct, others interpret correlations of the same magnitude as evidence for this. A less arbitrary way of deciding this would be to identify common mechanisms that underlie all kinds of perspective-taking and to examine whether manipulating these mechanisms in psychological experiments affects measures of perceptual, affective, and cognitive perspective-taking in parallel.
In accordance with this reasoning, the present dissertation assumes that the mental self-rotation of the body schema into the physical location of another person, the main mechanism of perceptual perspective-taking, is a common mechanism of all kinds of perspective-taking. Thus, contrary to previous research a unitary construct is not only justified on the grounds of a common central functionality of all kinds of perspective-taking, that is, overcoming one’s egocentrism in favor of an alternative (perceptual, affective or cognitive) point of view, but additionally on the grounds of a common psychological mechanism. From this, the simple hypothesis that inducing visuo-spatial perspective-taking also leads to psychological consequences is derived. This hypothesis was tested in 6 experiments.
In these experiments, participants first had to adopt the visual perspective of another person. To this end, they saw a person sitting at a table with two objects. During every trial, participants had to decide which hand the person would have to use in order to grab one of the two objects. Furthermore, the angular disparity between the participant and the target was manipulated in such a way that during half of the trials the target person was within the same visuo-spatial reference frame as the participant and thus no perspective-taking was necessary to solve the task correctly. During the remaining trials, the target person was sitting in another visuo-spatial reference frame so that the participants had to engage in perspective-taking to solve the task correctly. After every such trial, the target person was imbued with a mental state. This was done using an adapted paradigm for the investigation of the anchoring heuristic. Specifically, participants were asked to answer a trivia question and also saw what the target person from the visuo-spatial perspective-taking task was guessing.
In line with the hypothesis that visuo-spatial perspective-taking leads to psychological outcomes, too, it was found that participants adopted the thoughts of the target person more strongly after visuo-spatial perspective-taking. This was evident in the absolute size of the anchoring effect, as well as the differences between participant and target estimations. Further experiments ruled out sample and stimulus characteristics and task difficulty as alternative explanations for these effects. The last two experiments furthermore established that the effects were specific to constellations where an embodied self-rotation into the target’s perspective was necessary and that the adoption of the target’s thoughts was associated with feelings of similarity.
Taken together, these findings support the theoretically elaborated unitary view of perspective-taking and furthermore distinguish this construct from other related phenomena. In the general discussion, the significance of these findings for research on empathy, theory of mind, and perspective-taking, as well as practical implications are discussed.
BACKGROUND:
Thigmotaxis refers to a specific behavior of animals (i.e., to stay close to walls when exploring an open space). Such behavior can be assessed with the open field test (OFT), which is a well-established indicator of animal fear. The detection of similar open field behavior in humans may verify the translational validity of this paradigm. Enhanced thigmotaxis related to anxiety may suggest the relevance of such behavior for anxiety disorders, especially agoraphobia.
METHODS:
A global positioning system was used to analyze the behavior of 16 patients with agoraphobia and 18 healthy individuals with a risk for agoraphobia (i.e., high anxiety sensitivity) during a human OFT and compare it with appropriate control groups (n = 16 and n = 19). We also tracked 17 patients with agoraphobia and 17 control participants during a city walk that involved walking through an open market square. RESULTS: Our human OFT triggered thigmotaxis in participants; patients with agoraphobia and participants with high anxiety sensitivity exhibited enhanced thigmotaxis. This behavior was evident in increased movement lengths along the wall of the natural open field and fewer entries into the center of the field despite normal movement speed and length. Furthermore, participants avoided passing through the market square during the city walk, indicating again that thigmotaxis is related to agoraphobia.
CONCLUSIONS:
This study is the first to our knowledge to verify the translational validity of the OFT and to reveal that thigmotaxis, an evolutionarily adaptive behavior shown by most species, is related to agoraphobia, a pathologic fear of open spaces, and anxiety sensitivity, a risk factor for agoraphobia.
Animal models reflective of ulcerative colitis (UC) remain a major challenge, and yet are crucial to understand mechanisms underlying the onset of disease and inflammatory characteristics of relapses and remission. Mouse models in which colitis-like symptoms are induced through challenge with toxins such as oxazolone, dextran sodium sulfate (DSS) or 2,4,6-trinitrobenzenesulfonic acid (TNBS) have been instrumental in understanding the inflammatory processes of UC. However, these neither reflect the heterogeneous symptoms observed in the UC-affected population nor can they be used to test the efficacy of inhibitors developed against human targets where high sequence and structural similarity of the respective ligands is lacking. In an attempt to overcome these problems, we have developed a mouse model that relies on NOD-scid IL2R γnull mice reconstituted with peripheral blood mononuclear cells derived from UC-affected individuals. Upon challenge with ethanol, mice developed colitis-like symptoms and changes in the colon architecture, characterized by influx of inflammatory cells, edema, crypt loss, crypt abscesses and epithelial hyperplasia, as previously observed in immune-competent mice. TARC, TGFβ1 and HGF expression increased in distal parts of the colon. Analysis of human leucocytes isolated from mouse spleen revealed an increase in frequencies of CD1a+, CD64+, CD163+ and TSLPR+ CD14+ monocytes, and antigen-experienced CD44+ CD4+ and CD8+ T-cells in response to ethanol. Analysis of human leucocytes from the colon of challenged mice identified CD14+ monocytes and CD11b+ monocytes as the predominant populations. Quantitative real-time PCR (RT-PCR) analysis from distal parts of the colon indicated that IFNγ might be one of the cytokines driving inflammation. Treatment with infliximab ameliorated symptoms and pathological manifestations, whereas pitrakinra had no therapeutic benefit. Thus, this model is partially reflective of the human disease and might help to increase the translation of animal and clinical studies.
Chronobiological studies of individual activity rhythms in social insects can be constrained by the artificial isolation of individuals from their social context. We present a new experimental set-up that simultaneously measures the temperature rhythm in a queen-less but brood raising mini colony and the walking activity rhythms of singly kept honey bees that have indirect social contact with it. Our approach enables monitoring of individual bees in the social context of a mini colony under controlled laboratory conditions. In a pilot experiment, we show that social contact with the mini colony improves the survival of monitored young individuals and affects locomotor activity patterns of young and old bees. When exposed to conflicting Zeitgebers consisting of a light-dark (LD) cycle that is phase-delayed with respect to the mini colony rhythm, rhythms of young and old bees are socially synchronized with the mini colony rhythm, whereas isolated bees synchronize to the LD cycle. We conclude that the social environment is a stronger Zeitgeber than the LD cycle and that our new experimental set-up is well suited for studying the mechanisms of social entrainment in honey bees.
G-protein coupled receptors (GPCRs) can activate a heterotrimeric G-protein complex with subsecond kinetics. Genetically encoded biosensors based on Förster resonance energy transfer (FRET) are ideally suited for the study of such fast signaling events in single living cells. Here we report on the construction and characterization of three FRET biosensors for the measurement of Gα\(_{i1}\), Gα\(_{i2}\) and Gα\(_{i3}\) activation. To enable quantitative long-term imaging of FRET biosensors with high dynamic range, fluorescent proteins with enhanced photophysical properties are required. Therefore, we use the currently brightest and most photostable CFP variant, mTurquoise2, as donor fused to Gα\(_{i}\) subunit, and cp173Venus fused to the Gγ\(_{2}\) subunit as acceptor. The Gα\(_{i}\) FRET biosensors constructs are expressed together with Gβ\(_{1}\) from a single plasmid, providing preferred relative expression levels with reduced variation in mammalian cells. The Gα\(_{i}\) FRET sensors showed a robust response to activation of endogenous or over-expressed alpha-2A-adrenergic receptors, which was inhibited by pertussis toxin. Moreover, we observed activation of the Gα\(_{i}\) FRET sensor in single cells upon stimulation of several GPCRs, including the LPA\(_{2}\), M\(_{3}\) and BK\(_{2}\) receptor. Furthermore, we show that the sensors are well suited to extract kinetic parameters from fast measurements in the millisecond time range. This new generation of FRET biosensors for Gα\(_{i1}\), Gα\(_{i2}\) and Gα\(_{i3}\) activation will be valuable for live-cell measurements that probe Gα\(_{i}\) activation.
Motivation:
Next generation sequencing technologies have provided us with a wealth of information on genetic variation, but predi cting the functional significance of this variation is a difficult task. While many comparative genomics studies have focused on gene flux and large scale changes, relatively little attention has been paid to quantifying the effects of single nucleotide polymorphisms and indels on protein function, particularly in bacterial genomics.
Results:
We present a hidden Markov model based approach we call delta-bitscore (DBS) for identifying orthologous proteins that have diverged at the amino acid sequence level in a way that is likely to impact biological function. We benchmark this approach with several widely used datasets and apply it to a proof-of-concept study of orthologous proteomes in an investigation of host adaptation in Salmonella enterica. We highlight the value of the method in identifying functional divergence of genes, and suggest that this tool may be a better approach than the commonly used dN/dS metric for identifying functionally significant genetic changes occurring in recently diverged organisms.
The most energetic versions of active galactic nuclei (AGNs) feature two highly-relativistic plasma outflows, so-called jets, that are created in the vicinity of the central supermassive black hole and evolve in opposite directions. In blazars, which dominate the extragalactic gamma-ray sky, the jets are aligned close to the observer's line of sight leading to strong relativistic beaming effects of the jet emission. Radio observations especially using very long baseline interferometry (VLBI) provide the best way to gain direct information on the intrinsic properties of jets down to sub-parsec scales, close to their formation region.
In this thesis, I focus on the properties of three AGNs, IC 310, PKS 2004-447, and 3C 111 that belong to the small non-blazar population of gamma-ray-loud AGNs. In these kinds of AGNs, the jets are less strongly aligned with respect to the observer than in blazars. I study them in detail with a variety of radio astronomical instruments with respect to their high-energy emission and in the context of the large samples in the monitoring programmes MOJAVE and TANAMI. My analysis of radio interferometric observations and flux density monitoring data reveal very different characteristics of the jet emission in these sources. The work presented in this thesis illustrates the diversity of the radio properties of gamma-ray-loud AGNs that do not belong to the dominating class of blazars.
Olfactory receptors (ORs) are G protein-coupled receptors which serve important sensory functions beyond their role as odorant detectors in the olfactory epithelium. Here we describe a novel role for one of these ORs, Olfr1393, as a regulator of renal glucose handling. Olfr1393 is specifically expressed in the kidney proximal tubule, which is the site of renal glucose reabsorption. Olfr1393 knockout mice exhibit urinary glucose wasting and improved glucose tolerance, despite euglycemia and normal insulin levels. Consistent with this phenotype, Olfr1393 knockout mice have a significant decrease in luminal expression of Sglt1, a key renal glucose transporter, uncovering a novel regulatory pathway involving Olfr1393 and Sglt1. In addition, by utilizing a large scale screen of over 1400 chemicals we reveal the ligand profile of Olfr1393 for the first time, offering new insight into potential pathways of physiological regulation for this novel signaling pathway.