Partial (13)C isotopic enrichment of nucleoside monophosphates: useful reporters for NMR structural studies

Analysis of the (13)C isotopic labeling patterns of nucleoside monophosphates (NMPs) extracted from Escherichia coli grown in a mixture of C-1 and C-2 glucose is presented. By comparing our results to previous observations on amino acids grown in similar media, we have been able to rationalize the labeling pattern based on the well-known biochemistry of nucleotide biosynthesis. Except for a few notable absences of label (C4 in purines and C3′ in ribose) and one highly enriched site (C1′ in ribose), most carbons are randomly enriched at a low level (an average of 13%). These sparsely labeled NMPs give less complex NMR spectra than their fully isotopically labeled analogs due to the elimination of most (13)C–(13)C scalar couplings. The spectral simplicity is particularly advantageous when working in ordered systems, as illustrated with guanosine diphosphate (GDP) bound to ADP ribosylation factor 1 (ARF1) aligned in a liquid crystalline medium. In this system, the absence of scalar couplings and additional long-range dipolar couplings significantly enhances signal to noise and resolution

Physical properties of seven binary and higher-order multiple OB systems

Analyses of multi-epoch, high-resolution (~ 50000) optical spectra of seven early-type systems provided various important new insights with respect to their multiplicity. First determinations of orbital periods were made for HD 92206C (2.022 d), HD 112244 (27.665 d), HD 164438 (10.25 d), HD 123056A (~ 1314 d) and HD 123056B (< 2 d); the orbital period of HD 318015 could be improved (23.445975 d). Concerning multiplicity, a third component was discovered for HD 92206C by means of He I line profiles. For HD 93146A, which was hitherto assumed to be SB1, lines of a secondary component could be discerned. HD 123056 turns out to be a multiple system consisting of a high-mass component A (~ O8.5) displaying a broad He II 5411 A feature with variable radial velocity, and of an inner pair B (~ B0) with double He I lines. The binary HD 164816 was revisited and some of its system parameters were improved. In particular, we determined its systemic velocity to be -7 km/s, which coincides with the radial velocity of the cluster NGC 6530. This fact, together with its distance, suggests the cluster membership of HD 164816. The OB system HD 318015 (V1082 Sco) belongs to the rare class of eclipsing binaries with a supergiant primary (B0.5/0.7). Our combined orbital and light-curve analysis suggests that the secondary resembles an O9.5III star. Our results for a limited sample corroborate the findings that many O stars are actually massive multiple systems.Comment: 16 pages, 16 figures, to appear in Astronomy and Astrophysic

Unveiling the Subjective Perception of Health Information Sources: A Three-Dimensional Source Taxonomy Based on Similarity Judgements

Although there is a multitude of taxonomies of health information sources, these taxonomies only partly include how information users classify these sources. The present paper complements this research by developing a taxonomy which is based on individuals' subjective perceptions of the “universe” of health information sources. In our study, nine non-redundant sources of health information were presented to N = 150 participants who rated all 36 possible combinations of source pairs regarding their perceived similarity. Results of non-metric multidimensional scaling suggested three basic dimensions underlying the similarity ratings: “expertise” (lay vs. professional), “interaction” (interpersonal vs. impersonal), and “accessibility” (low vs. high). Thus, the wide array of health information sources can be structured by means of the new taxonomy arranging them on three dimensions. This allows researchers to classify each source and with this, to draw on common ground when interpreting the varying use of health information sources

Small town research in Germany - status quo and recommendations

Urban studies in Germany are traditionally oriented towards large cities. The structures, meanings and functions of small towns are not sufficiently perceived and differentiated in scientific or political debates. Adequate research on small towns requires systematic, comparative, inter- and transdisciplinary approaches. Traditional attributions should be questioned critically and small towns should be examined empirically in their diversity and differentiation. This involves paying attention to external influences and heterogeneous internal structures as well as to regional functions and interdependencies. The availability and generation of statistical data, which also make small-scale analyses possible, are just as necessary as more comprehensive studies, which go beyond limited case studies. Finally, also research funding and academic teaching should address small towns more systematically than it has been the case in the past. This position paper presents recommendations for research, university teaching, official statistics and research funding in the field of small town research. The Ad-hoc Working Group focused on small town research in Germany and German-language literature, respectively

No sex scandals please, we're French: French attitudes towards politicians' public and private conduct

The notion of distinct ‘public’ and ‘private’ spheres underpins much normative and practical engagement with political misconduct. What is less clear is whether citizens draw distinctions between misdemeanours in the ‘public’ and ‘private’ spheres, and whether they judge these in systematically different ways. This paper explores attitudes to political misconduct in France. French citizens are often said to be particularly relaxed about politicians’ private affairs, but there has been little empirical evidence for this proposition. Drawing on original survey data, this paper demonstrates clearly that French citizens draw a sharp distinction between politicians’ public and private transgressions, and are more tolerant of the latter

Spectral properties of zero temperature dynamics in a model of a compacting granular column

The compacting of a column of grains has been studied using a one-dimensional Ising model with long range directed interactions in which down and up spins represent orientations of the grain having or not having an associated void. When the column is not shaken (zero 'temperature') the motion becomes highly constrained and under most circumstances we find that the generator of the stochastic dynamics assumes an unusual form: many eigenvalues become degenerate, but the associated multi-dimensional invariant spaces have but a single eigenvector. There is no spectral expansion and a Jordan form must be used. Many properties of the dynamics are established here analytically; some are not. General issues associated with the Jordan form are also taken up.Comment: 34 pages, 4 figures, 3 table

Rapid conditional knock-down–knock-in system for mammalian cells

RNA interference (RNAi) is a powerful tool to analyze gene function in mammalian cells. However, the interpretation of RNAi knock-down phenotypes can be hampered by off-target effects or compound phenotypes, as many proteins combine multiple functions within one molecule and coordinate the assembly of multimolecular complexes. Replacing the endogenous protein with ectopic wild-type or mutant forms can exclude off-target effects, preserve complexes and unravel specific roles of domains or modifications. Therefore, we developed a rapid-knock-down–knock-in system for mammalian cells. Stable polyclonal cell lines were generated within 2 weeks by simultaneous selection of two episomal vectors. Together these vectors mediated reconstitution and knock-down in a doxycycline-dependent manner to allow the analysis of essential genes. Depletion was achieved by an artificial miRNA-embedded siRNA targeting the untranslated region of the endogenous, but not the ectopic mRNA. To prove effectiveness, we tested 17 mutants of WDR12, a factor essential for ribosome biogenesis and cell proliferation. Loss-off function phenotypes were rescued by the wild-type and six mutant forms, but not by the remaining mutants. Thus, our system is suitable to exclude off-target effects and to functionally analyze mutants in cells depleted for the endogenous protein

Elevated Pre-Treatment Serum MMP-7 Levels Are Associated with the Presence of Metastasis and Poor Survival in Upper Tract Urothelial Carcinoma

Upper tract urothelial carcinoma (UTUC) is a rare cancer with a barely predictable clinical behaviour. Serum MMP-7 is a validated prognostic marker in urothelial bladder cancer, a tumour entity with large clinical, histological, and molecular similarity to UTUC. The serum MMP-7 levels have not yet been investigated in UTUC. In the present study, we determined MMP-7 concentrations in an overall number of 103 serum samples from 57 UTUC patients who underwent surgical or systemic (platinum or immune checkpoint inhibitor) therapy by using the ELISA method. In addition to pre-treatment samples, the serum samples collected at predefined time points after or during therapy were also investigated. Serum MMP-7 concentrations were correlated with clinicopathological and follow-up data. Our results revealed significantly, two-fold elevated pre-treatment serum MMP-7 levels in metastatic cases of UTUC in both the radical surgery- and the chemotherapy-treated cohorts (p = 0.045 and p = 0.040, respectively). In addition, high serum MMP-7 levels significantly decreased after radical surgery, and high pre-treatment MMP-7 concentrations were associated with shorter survival both in the surgery- and chemotherapy-treated cohorts (p = 0.029 and p = 0.001, respectively). Our results revealed pre-treatment serum MMP-7 as a prognostic marker for UTUC, which may help to improve preoperative risk-stratification and thereby improve therapeutic decision-making