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Fedders, Ronja ; Muenzner, Matthias ; Weber, Pamela ; Sommerfeld, Manuela ; Knauer, Miriam ; Kedziora, Sarah ; Kast, Naomi ; Heidenreich, Steffi ; Raila, Jens ; Weger, Stefan ; Henze, Andrea ; Schupp, Michael
Retinol-binding protein 4 (RBP4) is the major transport protein for retinol in blood. Recent evidence from genetic mouse models shows that circulating RBP4 derives exclusively from hepatocytes. Because RBP4 is elevated in obesity and associates with the development of glucose intolerance and insulin resistance, we tested whether a liver-specific overexpression of RBP4 in mice impairs glucose homeostasis. We used adeno-associated viruses (AAV) that contain a highly liver-specific promoter to drive expression of murine RBP4 in livers of adult mice. The resulting increase in serum RBP4 levels in these mice was comparable with elevated levels that were reported in obesity. Surprisingly, we found that increasing circulating RBP4 had no effect on glucose homeostasis. Also during a high-fat diet challenge, elevated levels of RBP4 in the circulation failed to aggravate the worsening of systemic parameters of glucose and energy homeostasis. These findings show that liver-secreted RBP4 does not impair glucose homeostasis. We conclude that a modest increase of its circulating levels in mice, as observed in the obese, insulin-resistant state, is unlikely to be a causative factor for impaired glucose homeostasis.
Leveraging large-deviation statistics to decipher the stochastic properties of measured trajectories
(2021)
Thapa, Samudrajit ; Wyłomańska, Agnieszka ; Sikora, Grzegorz ; Wagner, Caroline E. ; Krapf, Diego ; Kantz, Holger ; Chechkin, Aleksei V. ; Metzler, Ralf
Extensive time-series encoding the position of particles such as viruses, vesicles, or individualproteins are routinely garnered insingle-particle tracking experiments or supercomputing studies.They contain vital clues on how viruses spread or drugs may be delivered in biological cells.Similar time-series are being recorded of stock values in financial markets and of climate data.Such time-series are most typically evaluated in terms of time-averaged mean-squareddisplacements (TAMSDs), which remain random variables for finite measurement times. Theirstatistical properties are different for differentphysical stochastic processes, thus allowing us toextract valuable information on the stochastic process itself. To exploit the full potential of thestatistical information encoded in measured time-series we here propose an easy-to-implementand computationally inexpensive new methodology, based on deviations of the TAMSD from itsensemble average counterpart. Specifically, we use the upper bound of these deviations forBrownian motion (BM) to check the applicability of this approach to simulated and real data sets.By comparing the probability of deviations fordifferent data sets, we demonstrate how thetheoretical bound for BM reveals additional information about observed stochastic processes. Weapply the large-deviation method to data sets of tracer beads tracked in aqueous solution, tracerbeads measured in mucin hydrogels, and of geographic surface temperature anomalies. Ouranalysis shows how the large-deviation properties can be efficiently used as a simple yet effectiveroutine test to reject the BM hypothesis and unveil relevant information on statistical propertiessuch as ergodicity breaking and short-time correlations.
Water-deficits can cause lethal damage to organisms, which is rooted in cellular dehydration. Many plant species, but also other organisms have developed mechanisms to tolerate such stresses, such as the expression of LEA proteins. Many studies report on physiological protective functions of LEA proteins but lack information about their precise mechanisms on a molecular level. Most LEA proteins are intrinsically disordered in dilute solution but may adopt a distinct secondary structure upon changes in solvent conditions. Understanding the molecular mechanism of how LEA proteins contribute to the counteraction of cellular damage during water-deficits may in the long-term pave the way for breeding crops that are resistant to the effects of global warming. The objective of the work at hand is to improve the biophysical understanding of the sequencestructure-function relationship of LEA proteins as membrane stabilizers, based on the LEA_4 family of the model plant A. thaliana. This is pursued by using a combination of spectroscopic and scattering techniques, supported by bioinformatics and computational analyses. Eight out of the 18 LEA_4 proteins are experimentally assessed revealing that a coil-helix transition in response to water-deficit is a common feature, as predicted for the entire family. In addition, they all stabilize simple membrane models during a freeze/ thaw cycle. Three-dimensional structure prediction of representative members suggests that their completely folded states are represented by a sequential arrangement of alpha-helical segments connected by unstructured linkers, which is experimentally verified for the LEA_4 protein COR15A. The unstructured linker region of COR15A represents a conserved motif among its closest homologs and is, therefore, of particular interest. Facilitating a set of seven designed and investigated COR15A mutants uncovers a complex interplay of transient interactions between the amphipathic alpha-helical segments, mediated by the linker, which fine-tunes folding transitions and structural ensembles upon reduced water-availability. Finally, alpha-helicity is also induced in COR15A upon temperature decrease, which is enhanced in the presence of osmolytes. In addition, high solution osmolarity induced secondary structure is followed by oligomerization of COR15A. Interestingly, the functionality of COR15A, in terms of liposome stabilization, strongly correlates with its alpha-helix ratio in the folded state. The present work significantly improves the understanding of the sequence-structure-function relationship for LEA_4 proteins and offers novel findings on folding mechanisms and oligomerization of COR15A.
The trace elements zinc and manganese are essential for human health, especially due to their enzymatic and protein stabilizing functions. If these elements are ingested in amounts exceeding the requirements, regulatory processes for maintaining their physiological concentrations (homeostasis) can be disturbed. Those homeostatic dysregulations can cause severe health effects including the emergence of neurodegenerative disorders such as Parkinson’s disease (PD). The concentrations of essential trace elements also change during the aging process. However, the relations of cause and consequence between increased manganese and zinc uptake and its influence on the aging process and the emergence of the aging-associated PD are still rarely understood. This doctoral thesis therefore aimed to investigate the influence of a nutritive zinc and/or manganese oversupply on the metal homeostasis during the aging process. For that, the model organism Caenorhabditis elegans (C. elegans) was applied. This nematode suits well as an aging and PD model due to properties such as its short life cycle and its completely sequenced, genetically amenable genome. Different protocols for the propagation of zinc- and/or manganese-supplemented young, middle-aged and aged C. elegans were established. Therefore, wildtypes, as well as genetically modified worm strains modeling inheritable forms of parkinsonism were applied. To identify homeostatic and neurological alterations, the nematodes were investigated with different methods including the analysis of total metal contents via inductively-coupled plasma tandem mass spectrometry, a specific probe-based method for quantifying labile zinc, survival assays, gene expression analysis as well as fluorescence microscopy for the identification and quantification of dopaminergic neurodegeneration.. During aging, the levels of iron, as well as zinc and manganese increased.. Furthermore, the simultaneous oversupply with zinc and manganese increased the total zinc and manganese contents to a higher extend than the single metal supplementation. In this relation the C. elegans metallothionein 1 (MTL-1) was identified as an important regulator of metal homeostasis. The total zinc content and the concentration of labile zinc were age-dependently, but differently regulated. This elucidates the importance of distinguishing these parameters as two independent biomarkers for the zinc status. Not the metal oversupply, but aging increased the levels of dopaminergic neurodegeneration. Additionally, nearly all these results yielded differences in the aging-dependent regulation of trace element homeostasis between wildtypes and PD models. This confirms that an increased zinc and manganese intake can influence the aging process as well as parkinsonism by altering homeostasis although the underlying mechanisms need to be clarified in further studies.
Breslau has been almost entirely forgotten in the Anglophone sphere as a place of Enlightenment. Moreover, in the context of the Jewish Enlightenment, Breslau has never been discussed as a place of intercultural exchange between German-speaking Jewish, Protestant and Catholic intellectuals. The story of Moses Hirschel offers us an excellent case-study to investigate the complex reciprocal relationship between Jewish and non-Jewish enlighteners in a prosperous and influential Central European city on the cusp of the 18th century.
Coordinated subjects often show variable number agreement with the finite verb, but linguistic approaches to this phenomenon have rarely been informed by systematically collected data. We report the results from three experiments investigating German speakers' agreement preferences with complex subjects joined by the correlative conjunctions sowohl horizontal ellipsis als auch ('both horizontal ellipsis and'), weder horizontal ellipsis noch ('neither horizontal ellipsis nor') or entweder horizontal ellipsis oder ('either horizontal ellipsis or'). We examine to what extent conjunction type and a conjunct's relative proximity to the verb affect the acceptability and processibility of singular vs. plural agreement. Experiment 1 was an untimed acceptability rating task, Experiment 2 a timed sentence completion task, and Experiment 3 was a self-paced reading task. Taken together, our results show that number agreement with correlative coordination in German is primarily determined by a default constraint triggering plural agreement, which interacts with linear order and semantic factors. Semantic differences between conjunctions only affected speakers' agreement preferences in the absence of processing pressure but not their initial agreement computation. The combined results from our offline and online experimental measures of German speakers' agreement preferences suggest that the constraints under investigation do not only differ in their relative weighting but also in their relative timing during agreement computation.
A convenient method for the synthesis of gamma-spirolactams in only three steps is described. Birch reduction of inexpensive and commercially available aromatic carboxylic acids in the presence of chloroacetonitrile affords nitriles in moderate to good yields. Suitable precursors are methyl-substituted benzoic acids, naphthoic, and anthroic acid. Subsequent catalytic hydrogenation proceeds smoothly with PtO2 or Raney Ni as catalysts and lactams are isolated in excellent yields and stereoselectivities. Thus, up to 3 new stereogenic centers can be constructed as sole diastereomers from achiral benzoic acids. Furthermore, it is possible to control the degree of saturation at different pressures, affording products with 0, 1, or 2 double bonds. Overall, more than 15 new gamma-spirolactams have been synthesized in analytically pure form.
Influence of functional groups on the ene reaction of singlet oxygen with 1,4-cyclohexadienes
(2021)
Fudickar, Werner ; Metz, Melanie ; Mai-Linde, Yasemin ; Krüger, Tobias ; Kelling, Alexandra ; Sperlich, Eric ; Linker, Torsten
The photooxygenation of 1,4-cyclohexadienes has been studied with a special focus on regio- and stereoselectivities. In all examples, only the methyl-substituted double bond undergoes an ene reaction with singlet oxygen, to afford hydroperoxides in moderate to good yields. We explain the high regioselectivities by a "large-group effect" of the adjacent quaternary stereocenter. Nitriles decrease the reactivity of singlet oxygen, presumably by quenching, but can stabilize proposed per-epoxide intermediates by polar interactions resulting in different stereoselectivities. Spiro lactams and lactones show an interesting effect on regio- and stereoselectivities of the ene reactions. Thus, singlet oxygen attacks the double bond preferentially anti to the carbonyl group, affording only one regioisomeric hydroperoxide. If the reaction occurs from the opposite face, the other regioisomer is exclusively formed by severe electrostatic repulsion in a perepoxide intermediate. We explain this unusual behavior by the fixed geometry of spiro compounds and call it a "spiro effect" in singlet oxygen ene reactions.
In the absence of a stranded auxiliary or modal, VP-topicalization in most Germanic languages gives rise to the presence of a dummy verb meaning 'do'. Cross-linguistically, this is a rather uncommon strategy as comparable VP-fronting constructions in other languages, e.g. Hebrew, Polish, and Portuguese, among many others, exhibit verb doubling. A comparison of several recent approaches to verb doubling in VP-fronting reveals that it is the consequence of VP-evacuating head movement of the verb to some higher functional head, which saves the (low copy of the) verb from undergoing copy deletion as part of the low VP copy in the VP-topicalization dependency. Given that almost all Germanic languages have such V-salvaging head movement, namely V-to-C movement, but do not show verb doubling, this paper suggests that V-raising is exceptionally impossible in VP-topicalization clauses and addresses the question of why it is blocked. After discussing and rejecting some conceivable explanations for the lack of verb doubling, I propose that the blocking effect arises from a bleeding interaction between V-to-C movement and VP-to-SpecCP movement. As both operations are triggered by the same head, i.e. C, the VP is always encountered first by a downward search algorithm. Movement of VP then freezes it and its lower copies for subextraction precluding subsequent V-raising. Crucially, this implies that there is no V-to-T raising in most Germanic languages. V2 languages with V-to-T raising, e.g. Yiddish, are correctly predicted to not exhibit the blocking effect.