Mapping of the insulin-like growth factor II binding site of the Type I insulin-like growth factor receptor by alanine scanning mutagenesis

AbstractThe Type I insulin-like growth factor receptor is a physiological receptor for insulin-like growth factor II (IGF-II). To characterize the molecular basis of the receptor's ligand binding properties, we have examined the effects of alanine mutations of residues in the ligand binding site of the receptor on its affinity for IGF-II. The functional epitope for IGF-II comprises residues in the N-terminal L1 domain and residues at the C-terminus of the α subunit. Cysteine rich domain residues do not appear to be critical for IGF-II binding

Microbiology of the stalactites from Grotta dei Cervi, Porto Badisco, Italy

The active stalactites from Grotta dei Cervi, Porto Badisco, southeastern Italy, were sampled to investigate the microbial communities present in these speleothems. Sampling was carried out in a transect about 150 m long in the central gallery, where numerous Gram-positive bacteria were isolated. Actinomycetes of the genus Streptomyces were the most abundant, followed by members of the genus Bacillus. Further isolates were assigned to the genera Amycolatopsis, Arthrobacter, Agromyces, Micrococcus, Nocardiopsis and Rhodococcus of the order Actinomycetales. The ability of actinomycetes to colonize subterranean environments is discussed

Klimaschutz in der räumlichen Planung : Gestaltungsmöglichkeiten der Raumordnung und Bauleitplanung ; Kurzdokumentation der Fallstudien

Als Grundlage für die Praxishilfe "Klimaschutz in der räumlichen Planung" wurde im Auftrag des Umweltbundesamtes eine Reihe von Konzepten, Plänen und Programmen auf teil- bzw. gesamtstädtischer, regionaler und Landesebene untersucht, die für die Integration des Klimaschutz in die räumliche Planung beispielhaft sind. Es handelt sich im Schwerpunkt um formelle Instrumente (Bebauungs- und Flächennutzungspläne, Regionalpläne, Landesentwicklungspläne). Komplementär wurden auch informelle Instrumente untersucht, die die fachliche Grundlage für eine integrierte und effektive Bearbeitung des Klimaschutzes in der förmlichen Raumplanung darstellen (v.a. Energieversorgungs- und Klimaschutzkonzepte)

Two distinct modes for propagation of histone PTMs across the cell cycle

Epigenetic states defined by chromatin can be maintained through mitotic cell division. However, it remains unknown how histone-based information is transmitted. Here we combine nascent chromatin capture (NCC) and triple-SILAC (stable isotope labeling with amino acids in cell culture) labeling to track histone modifications and histone variants during DNA replication and across the cell cycle. We show that post-translational modifications (PTMs) are transmitted with parental histones to newly replicated DNA. Di- and trimethylation marks are diluted twofold upon DNA replication, as a consequence of new histone deposition. Importantly, within one cell cycle, all PTMs are restored. In general, new histones are modified to mirror the parental histones. However, H3K9 trimethylation (H3K9me3) and H3K27me3 are propagated by continuous modification of parental and new histones because the establishment of these marks extends over several cell generations. Together, our results reveal how histone marks propagate and demonstrate that chromatin states oscillate within the cell cycle

Sobre la presència de Teucrium pumilum i Teucrium libanitis (Lamiaceae) a la província de València.

12p. fotog.color.[EN] About the presence of Teucrium pumilum and Teucrium libanitis (Lamiaceae) in the Valencian province, Spain.- Teucrium pumilum and T. libanitis have been cited from Valencia province (Spain) since the middle of the XXth Century from the gypsic soils in the Valle de Ayora-Cofrentes shire. The analysis of specimens and labels is not conclusive; no specimen would certify the presence of any of these taxa. The field identification and their inclusion in a phyotosociological table (relevé) by Rivas Goday are the only basis of their presence in the territory in a particular moment of the recent Spanish botanical history. Additionally, the unique herbarium specimen from Valencia, impossible to assign a concrete geographic locality, which was traditionally assigned to T. pumilum by some authors, is actually its congeneric T. carolipaui.[ES] Sobre la presencia de Teucrium pumilum y Teucrium libanitis (Lamiaceae) en la provincia de Valencia.- Teucrium pumilum y T. libanitis son dos especies que han sido citadas como presentes en la provincia de Valencia desde mediados del siglo XX, para los yesos que afloran en la comarca del Valle de Ayora-Cofrentes. El análisis de las etiquetas de los pliegos de los herbarios no es concluyente; ningún pliego testigo certificaría la presencia de estas especies. La determinación de visu por parte de Rivas Goday, y su inclusión en una tabla de inventarios fitosociológica, son el único fundamento de su existencia en la zona en algún momento determinado de la reciente historia botánica española. Además, para T. pumilum, el pliego de herbario que tradicionalmente se ha asignado a la cita valenciana se trata de una confusión en la determinación por parte de algunos autores con su congénere T. carolipaui, siendo imposible al mismo tiempo asignar una localidad geográfica concreta a este material.[CAT] Sobre la presència de Teucrium pumilum i Teucrium libanitis (Lamiaceae) a la província de València.- Teucrium pumilum i T. libanitis són dues espècies que han estat citades com a presents a la província de València des de mitjans del segle XX, sobre els guixos que afloren a la comarca de la Vall d’Aiora-Cofrents. L’anàlisi de les etiquetes dels plecs dels herbaris no és concloent; cap plec testimoni certificaria la presència d’aquestes espècies. La determinació de visu per part de Rivas Goday, i la seva inclusió en una taula d’inventaris fitosociològica, són l’únic fonament de la seva existència a la zona en un moment determinat de la recent història botànica espanyola. A més, per a T. pumilum, el plec d’herbari que tradicionalment s’ ha assignat a la cita valenciana es tracta d’una confusió en la determinació per part d’alguns autors amb el seu congènere.Peer reviewe

Haplotyping and copy number estimation of the highly polymorphic human beta-defensin locus on 8p23 by 454 amplicon sequencing

<p>Abstract</p> <p>Background</p> <p>The beta-defensin gene cluster (DEFB) at chromosome 8p23.1 is one of the most copy number (CN) variable regions of the human genome. Whereas individual DEFB CNs have been suggested as independent genetic risk factors for several diseases (e.g. psoriasis and Crohn's disease), the role of multisite sequence variations (MSV) is less well understood and to date has only been reported for prostate cancer. Simultaneous assessment of MSVs and CNs can be achieved by PCR, cloning and Sanger sequencing, however, these methods are labour and cost intensive as well as prone to methodological bias introduced by bacterial cloning. Here, we demonstrate that amplicon sequencing of pooled individual PCR products by the 454 technology allows in-depth determination of MSV haplotypes and estimation of DEFB CNs in parallel.</p> <p>Results</p> <p>Six PCR products spread over ~87 kb of DEFB and harbouring 24 known MSVs were amplified from 11 DNA samples, pooled and sequenced on a Roche 454 GS FLX sequencer. From ~142,000 reads, ~120,000 haplotype calls (HC) were inferred that identified 22 haplotypes ranging from 2 to 7 per amplicon. In addition to the 24 known MSVs, two additional sequence variations were detected. Minimal CNs were estimated from the ratio of HCs and compared to absolute CNs determined by alternative methods. Concordance in CNs was found for 7 samples, the CNs differed by one in 2 samples and the estimated minimal CN was half of the absolute in one sample. For 7 samples and 2 amplicons, the 454 haplotyping results were compared to those by cloning/Sanger sequencing. Intrinsic problems related to chimera formation during PCR and differences between haplotyping by 454 and cloning/Sanger sequencing are discussed.</p> <p>Conclusion</p> <p>Deep amplicon sequencing using the 454 technology yield thousands of HCs per amplicon for an affordable price and may represent an effective method for parallel haplotyping and CN estimation in small to medium-sized cohorts. The obtained haplotypes represent a valuable resource to facilitate further studies of the biomedical impact of highly CN variable loci such as the beta-defensin locus.</p

Effect of metal ions on the physical properties of multilayers from hyaluronan and chitosan, and the adhesion, growth and adipogenic differentiation of multipotent mouse fibroblasts

[EN] Polyelectrolyte multilayers (PEMs) consisting of the polysaccharides hyaluronic acid (HA) as the polyanion and chitosan (Chi) as the polycation were prepared with layer-by-layer technique (LbL). The [Chi/HA](5) multilayers were exposed to solutions of metal ions (Ca2+, Co2+, Cu2+ and Fe3+). Binding of metal ions to [Chi/HA](5) multilayers by the formation of complexes with functional groups of polysaccharides modulates their physical properties and the bioactivity of PEMs with regard to the adhesion and function of multipotent murine C3H10T1/2 embryonic fibroblasts. Characterization of multilayer formation and surface properties using different analytical methods demonstrates changes in the wetting, surface potential and mechanical properties of multilayers depending on the concentration and type of metal ion. Most interestingly, it is observed that Fe3+ metal ions greatly promote adhesion and spreading of C3H10T1/2 cells on the low adhesive [Chi/HA](5) PEM system. The application of intermediate concentrations of Cu2+, Ca2+ and Co2+ as well as low concentrations of Fe3+ to PEMs also results in increased cell spreading. Moreover, it can be shown that complex formation of PEMs with Cu2+ and Fe3+ ions leads to increased metabolic activity in cells after 24 h and induces cell differentiation towards adipocytes in the absence of any additional adipogenic media supplements. Overall, complex formation of [Chi/HA](5) PEM with metal ions like Cu2+ and Fe3+ represents an interesting and cheap alternative to the use of growth factors for making cell-adhesive coatings and guiding stem cell differentiation on implants and scaffolds to regenerate connective-type of tissues.This work was part of the High-Performance Center Chemical and Biosystems Technology Halle/Leipzig, supported by the European Regional Development Fund (ERDF), and a grant to HK from the Martin Luther University Halle-Wittenberg for female PhD students. The work was further supported by the Fraunhofer Internal Programs under Grant No. Attract 069-608203 (CEHS). TG acknowledges the kind support by the Ministry of Science and Higher Education of the Russian Federation within the framework of state support for the creation and development of World-Class Research Centers ``Digital biodesign and personalized healthcare'' 075-15-2020926. GGF acknowledges funding by the State Research Agency. Ministry of Science and Innovation of Spain, grant PID2019106000RB-C21/AEI/10.13039/501100011033 project. We are grateful for the kind support by Christian Willems for the help in formatting and proof reading the manuscript.Kindi, H.; Menzel, M.; Heilmann, A.; Schmelzer, CEH.; Herzberg, M.; Fuhrmann, B.; Gallego-Ferrer, G.... (2021). Effect of metal ions on the physical properties of multilayers from hyaluronan and chitosan, and the adhesion, growth and adipogenic differentiation of multipotent mouse fibroblasts. Soft Matter. 17(36):8394-8410. https://doi.org/10.1039/d1sm00405k83948410173