Effects of single and combined low frequency electromagnetic fields and simulated microgravity on gene expression of human mesenchymal stem cells during chondrogenesis

Introduction: Low frequency electromagnetic fields (LF-EMF) and simulated microgravity (SMG) have been observed to affect chondrogenesis. A controlled bioreactor system was developed to apply LF-EMF and SMG singly or combined during chondrogenic differentiation of human mesenchymal stem cells (hMSCs) in 3D culture. Material and methods: An external motor gear SMG bioreactor was combined with magnetic Helmholtz coils for EMF (5 mT;15 Hz). Pellets of hMSCs (+/- TGF-beta 3)were cultured (P5) under SMG, LF-EMF, LF-EMF/SMG and control (1 g) conditions for 3 weeks. Sections were stained with safranin-O and collagen type II. Gene expression was evaluated by microarray and real-time polymerase chain reaction analysis. Results: Simulated microgravity application significantly changed gene expression;specifically, COLXA1 but also COL2A1, which represents the chondrogenic potential, were reduced (p < 0.05). Low frequency electromagnetic fields application showed no gene expression changes on a microarray basis. LF-EMF/SMG application obtained significant different expression values from cultures obtained under SMG conditions with a re-increase of COL2A1, therefore rescuing the chondrogenic potential, which had been lowered by SMG. Conclusions: Simulated microgravity lowered hypertrophy but also the chondrogenic potential of hMSCs. Combined LF-EMF/SMG provided a rescue effect of the chondrogenic potential of hMSCs although no LF-EMF effect was observed under optimal conditions. The study provides new insights into how LF-EMF and SMG affect chondrogenesis of hMSCs and how they generate interdependent effects

Induction of sustained clinical remission in early axial spondyloarthritis following certolizumab pegol treatment: 48-week outcomes from C-OPTIMISE

INTRODUCTION: Achievement of remission is a key treatment goal for patients with axial spondyloarthritis (axSpA). C-OPTIMISE assessed achievement of sustained clinical remission in patients with axSpA, including radiographic (r) and non-radiographic (nr) axSpA, during certolizumab pegol (CZP) treatment, and subsequent maintenance of remission following CZP dose continuation, dose reduction or withdrawal. Here, we report outcomes from the first 48 weeks (induction period) of C-OPTIMISE, during which patients received open-label CZP. METHODS: C-OPTIMISE (NCT02505542) was a two-part, multicenter, phase 3b study in adult patients with early axSpA (r-/nr-axSpA), including a 48-week open-label induction period followed by a 48-week maintenance period. Patients with active adult-onset axSpA, < 5 years' symptom duration, and fulfilling Assessment of SpondyloArthritis international Society classification criteria, were included. During the induction period, patients received a loading dose of CZP 400 mg at weeks 0, 2, and 4, followed by CZP 200 mg every 2 weeks (Q2W) up to week 48. The main outcome of the 48-week induction period was the achievement of sustained clinical remission (defined as an Ankylosing Spondylitis Disease Activity Score [ASDAS] < 1.3 at week 32 and < 2.1 at week 36 [or vice versa], and < 1.3 at week 48). RESULTS: In total, 736 patients (407 with r-axSpA, 329 with nr-axSpA) were enrolled into the study. At week 48, 43.9% (323/736) of patients achieved sustained remission, including 42.8% (174/407) of patients with r-axSpA and 45.3% (149/329) with nr-axSpA. Patients also demonstrated substantial improvements in axSpA symptoms, MRI outcomes and quality of life measures. Adverse events occurred in 67.9% (500/736) of patients, of which 6.0% (44/736) were serious. CONCLUSIONS: Over 40% of patients with early axSpA achieved sustained remission during 48 weeks of open-label CZP treatment. Additionally, patients across the axSpA spectrum demonstrated substantial improvements in imaging outcomes and quality of life following treatment. No new safety signals were identified. TRIAL REGISTRATION: NCT02505542

Developmentally regulated GTP binding protein 1 (DRG1) controls microtubule dynamics

The mitotic spindle, essential for segregating the sister chromatids into the two evolving daughter cells, is composed of highly dynamic cytoskeletal filaments, the microtubules. The dynamics of microtubules are regulated by numerous microtubule associated proteins. We identify here Developmentally regulated GTP binding protein 1 (DRG1) as a microtubule binding protein with diverse microtubule-associated functions. In vitro, DRG1 can diffuse on microtubules, promote their polymerization, drive microtubule formation into bundles, and stabilize microtubules. HeLa cells with reduced DRG1 levels show delayed progression from prophase to anaphase because spindle formation is slowed down. To perform its microtubule-associated functions, DRG1, although being a GTPase, does not require GTP hydrolysis. However, all domains are required as truncated versions show none of the mentioned activities besides microtubule binding

A Rapid, Strong, and Convergent Genetic Response to Urban Habitat Fragmentation in Four Divergent and Widespread Vertebrates

Urbanization is a major cause of habitat fragmentation worldwide. Ecological and conservation theory predicts many potential impacts of habitat fragmentation on natural populations, including genetic impacts. Habitat fragmentation by urbanization causes populations of animals and plants to be isolated in patches of suitable habitat that are surrounded by non-native vegetation or severely altered vegetation, asphalt, concrete, and human structures. This can lead to genetic divergence between patches and in turn to decreased genetic diversity within patches through genetic drift and inbreeding.We examined population genetic patterns using microsatellites in four common vertebrate species, three lizards and one bird, in highly fragmented urban southern California. Despite significant phylogenetic, ecological, and mobility differences between these species, all four showed similar and significant reductions in gene flow over relatively short geographic and temporal scales. For all four species, the greatest genetic divergence was found where development was oldest and most intensive. All four animals also showed significant reduction in gene flow associated with intervening roads and freeways, the degree of patch isolation, and the time since isolation.Despite wide acceptance of the idea in principle, evidence of significant population genetic changes associated with fragmentation at small spatial and temporal scales has been rare, even in smaller terrestrial vertebrates, and especially for birds. Given the striking pattern of similar and rapid effects across four common and widespread species, including a volant bird, intense urbanization may represent the most severe form of fragmentation, with minimal effective movement through the urban matrix

WePass Störe am Eisernen Tor Wiederherstellung der Durchgängkeit an den Staustufen Iron Gate 1 & 2 in der Unteren Donau

Das Einzugsgebiet der 2.857 km langen Donau ist mit einer Gesamtfläche von 801.463 km² - rund 10% des europäischen Festlands - das zweitgrößte Flusseinzugsgebiet in Europa. Es erstreckt sich über 19 Länder und ist damit das internationalste Flusseinzugsgebiet der Welt. Über 100 Fischarten sind in der Donau nachgewiesen, darunter vier hochgradig gefährdete Störarten, die charakteristischen Flaggschiffarten der Donau (SCHIEMER ET AL. 2004). Die diadromen Donaufischarten (z. B. Störe (Acipenser spp.), Pontischer Maifisch (Alosa immaculata), Asowscher Maifisch (Alosa tanaica) und Schwarzmeerforelle (Salmo labrax)) sind zwingend darauf angewiesen ihren Lebenszyklus im Süß- und Salzwasser zu vollziehen. Einige Arten wandern bzw. wanderten dabei über sehr große Distanzen, beispielsweise der Beluga-Stör (Huso huso) vom Schwarzen Meer bis zu den Laichgründen >2.500 km ins Landesinnere (SCHMALL & FRIEDRICH 2014). Aber auch potamodrome Arten migrieren in der Donau über weite Strecken; für Döbel (Leuciscus cephalus), Barbe (Barbus barbus) und Nase (Chondrostoma nasus) sind z. B. Wanderungen über 169, 318 und 446 km nachgewiesen (STEINMANN ET AL. 1937, VAN TREECK ET AL. 2022). Die Fischartenzusammensetzung und der Fischbestand sind wichtige Indikatoren für die Gewässerqualität der Donau. Die Ergebnisse des jüngsten Joint Danube Survey 4 zeigen, dass die Fischartengemeinschaft entlang des gesamten Flusslaufs bedroht ist. Andererseits ist die Vielfalt der Fischtaxa immer noch auf einem Niveau, als dass wirksame Gewässerentwicklungsmaßnahmen dazu beitragen können den ökologischen Zustand bzw. das ökologische Potential zu verbessern und die Ziele der EG-WRRL zu erreichen (BĂNĂDUC ET AL. 2014, ICPDR 2021)

Search for the Pair Production of Dark Particles XX with KL0→XXK_L^0 \to XX, X→γγX \to \gamma\gamma

We present the first search for the pair production of dark particles $X$ via $K_L^0\to XX$ with $X$ decaying into two photons using the data collected by the KOTO experiment. No signal was observed in the mass range of 40 - 110~MeV/c$^2$ and 210 - 240 MeV/c$^2$. This sets upper limits on the branching fractions as $\mathcal{B}(K_L^0 \to XX)$ $<$ (1-4) $\times$ 10$^{-7}$ and $\mathcal{B}(K_L^0 \to XX)$ $<$ (1-2) $\times$ 10$^{-6}$ at the 90% confidence level for the two mass regions, respectively

Tubulin isoform composition tunes microtubule dynamics

Microtubules polymerize and depolymerize stochastically, a behavior essential for cell division, motility and differentiation. While many studies advanced our understanding of how microtubule-associated proteins tune microtubule dynamics in trans, we have yet to understand how tubulin genetic diversity regulates microtubule functions. The majority of in vitro dynamics studies are performed with tubulin purified from brain tissue. This preparation is not representative of tubulin found in many cell types. Here we report the 4.2Å cryo-EM structure and in vitro dynamics parameters of α1B/βI+βIVb microtubules assembled from tubulin purified from a human embryonic kidney cell line with isoform composition characteristic of fibroblasts and many immortalized cell lines. We find that these microtubules grow faster and transition to depolymerization less frequently compared to brain microtubules. Cryo-EM reveals that the dynamic ends of α1B/βI+βIVb microtubules are less tapered and that these tubulin heterodimers display lower curvatures. Interestingly, analysis of EB1 distributions at dynamic ends suggests no differences in GTP cap sizes. Lastly, we show that the addition of recombinant α1A/βIII tubulin, a neuronal isotype overexpressed in many tumors, proportionally tunes the dynamics of α1B/βI+βIVb microtubules. Our study is an important step towards understanding how tubulin isoform composition tunes microtubule dynamics

Site-Specific Bioconjugation of a Murine Dihydrofolate Reductase Enzyme by Copper(I)-Catalyzed Azide-Alkyne Cycloaddition with Retained Activity

Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) is an efficient reaction linking an azido and an alkynyl group in the presence of copper catalyst. Incorporation of a non-natural amino acid (NAA) containing either an azido or an alkynyl group into a protein allows site-specific bioconjugation in mild conditions via CuAAC. Despite its great potential, bioconjugation of an enzyme has been hampered by several issues including low yield, poor solubility of a ligand, and protein structural/functional perturbation by CuAAC components. In the present study, we incorporated an alkyne-bearing NAA into an enzyme, murine dihydrofolate reductase (mDHFR), in high cell density cultivation of Escherichia coli, and performed CuAAC conjugation with fluorescent azide dyes to evaluate enzyme compatibility of various CuAAC conditions comprising combination of commercially available Cu(I)-chelating ligands and reductants. The condensed culture improves the protein yield 19-fold based on the same amount of non-natural amino acid, and the enzyme incubation under the optimized reaction condition did not lead to any activity loss but allowed a fast and high-yield bioconjugation. Using the established conditions, a biotin-azide spacer was efficiently conjugated to mDHFR with retained activity leading to the site-specific immobilization of the biotin-conjugated mDHFR on a streptavidin-coated plate. These results demonstrate that the combination of reactive non-natural amino acid incorporation and the optimized CuAAC can be used to bioconjugate enzymes with retained enzymatic activityope

WEPASS Project - Making the Iron Gate Dams passable for migratory fish

European rivers are obstructed by more than one million barriers that have resulted in excessive loss of river continuity. On the main course of the Danube River there are 83 longitudinal continuity interruptions, out of which 65 dams are used for hydropower (ICPDR 2022a). The Iron Gate Hydropower and Navigation System is one of the largest river engineering projects undertaken in Europe, with the dams mainly built to provide hydropower and flood protection, and to facilitate navigation along the Danube. These infrastructures represent introduced barriers to fish migration. Hence, ensuring passage opportunities for fish at the Iron Gate dams is considered to be of major importance for the conservation of migratory fish populations in the Danube River basin. Restoration of river continuity at these sites would reopen an additional 900 km for migration up to the Gabčikovo dam, providing suitable habitats and spawning grounds along the Danube and its tributaries. Knowledge about fish behavior and movements in the vicinity of these river infrastructures is required to build effective up- and downstream passage facilities to allow the migration of fish species. To gain insight in the approach routes and aggregation areas a refined approach to acoustic telemetry is employed to support migration facilitation