Repression of anti-proliferative factor Tob1 in osteoarthritic cartilage

Osteoarthritis is the most common degenerative disorder of the modern world. However, many basic cellular features and molecular processes of the disease are poorly understood. In the present study we used oligonucleotide-based microarray analysis of genes of known or assumed relevance to the cellular phenotype to screen for relevant differences in gene expression between normal and osteoarthritic chondrocytes. Custom made oligonucleotide DNA arrays were used to screen for differentially expressed genes in normal (n = 9) and osteoarthritic (n = 10) cartilage samples. Real-time polymerase chain reaction (PCR) with gene-specific primers was used for quantification. Primary human adult articular chondrocytes and chondrosarcoma cell line HCS-2/8 were used to study changes in gene expression levels after stimulation with interleukin-1β and bone morphogenetic protein, as well as the dependence on cell differentiation. In situ hybridization with a gene-specific probe was applied to detect mRNA expression levels in fetal growth plate cartilage. Overall, more than 200 significantly regulated genes were detected between normal and osteoarthritic cartilage (P < 0.01). One of the significantly repressed genes, Tob1, encodes a protein belonging to a family involved in silencing cells in terms of proliferation and functional activity. The repression of Tob1 was confirmed by quantitative PCR and correlated to markers of chondrocyte activity and proliferation in vivo. Tob1 expression was also detected at a decreased level in isolated chondrocytes and in the chondrosarcoma cell line HCS-2/8. Again, in these cells it was negatively correlated with proliferative activity and positively with cellular differentiation. Altogether, the downregulation of the expression of Tob1 in osteoarthritic chondrocytes might be an important aspect of the cellular processes taking place during osteoarthritic cartilage degeneration. Activation, the reinitiation of proliferative activity and the loss of a stable phenotype are three major changes in osteoarthritic chondrocytes that are highly significantly correlated with the repression of Tob1 expression

Inhibition of iodine organification and regulation of follicular size in rat thyroid tissue in vitro

The factors mediating the accumulation of thyroglobulin are of great importance to the understanding of the pathogenesis of human and experimentally induced colloid goiters. To elucidate further the underlying cellular mechanism, thyroid fragments from newborn rats were incorporated into semisolid alginate beads and were cultured as three-dimensional organoids for up to 21 d. In five parallel cultures, the medium contained either no supplements (group A), Nal (group B), thyroid-stimulating hormone (TSH) (group C), Nal plus TSH in the same concentrations as B and C (group D), or Nal and TSH (as in group D) plus methimazole (MMI, group E). The thyroid organoids maintained morphological intergrity, functional activity, and ability to proliferate in vitro. Addition of iodine to the cultures significantly increased mean (±SEM) follicular diameters from 19.5±0.7 μm in controls to 33.9±2.2 μm (p<0.0001) when Nal was added alone (group B), and 30.4±1.7 μm (p<0.0001) when combined with TSH (group D). The effect of Nal on follicular size was abolished by MMI (group E, follicular diameter 23.5±1.3 μm). The results presented support the recent finding, using a rat colloid goiter model, that not only TSH but also iodine organification or its inhibition are important factors in modulating follicular morpholog

Correlational analysis for identifying genes whose regulation contributes to chronic neuropathic pain

<p>Abstract</p> <p>Background</p> <p>Nerve injury-triggered hyperexcitability in primary sensory neurons is considered a major source of chronic neuropathic pain. The hyperexcitability, in turn, is thought to be related to transcriptional switching in afferent cell somata. Analysis using expression microarrays has revealed that many genes are regulated in the dorsal root ganglion (DRG) following axotomy. But which contribute to pain phenotype versus other nerve injury-evoked processes such as nerve regeneration? Using the L5 spinal nerve ligation model of neuropathy we examined <b><it>differential </it></b>changes in gene expression in the L5 (and L4) DRGs in five mouse strains with contrasting susceptibility to neuropathic pain. We sought genes for which the degree of regulation correlates with strain-specific pain phenotype.</p> <p>Results</p> <p>In an initial experiment six candidate genes previously identified as important in pain physiology were selected for in situ hybridization to DRG sections. Among these, regulation of the Na⁺channel α subunit <it>Scn11a </it>correlated with levels of spontaneous pain behavior, and regulation of the cool receptor <it>Trpm8 </it>correlated with heat hypersensibility. In a larger scale experiment, mRNA extracted from individual mouse DRGs was processed on Affymetrix whole-genome expression microarrays. Overall, 2552 ± 477 transcripts were significantly regulated in the axotomized L5DRG 3 days postoperatively. However, in only a small fraction of these was the degree of regulation correlated with pain behavior across strains. Very few genes in the "uninjured" L4DRG showed altered expression (24 ± 28).</p> <p>Conclusion</p> <p>Correlational analysis based on in situ hybridization provided evidence that differential regulation of <it>Scn11a </it>and <it>Trpm8 </it>contributes to across-strain variability in pain phenotype. This does not, of course, constitute evidence that the others are unrelated to pain. Correlational analysis based on microarray data yielded a larger "look-up table" of genes whose regulation likely contributes to pain variability. While this list is enriched in genes of potential importance for pain physiology, and is relatively free of the bias inherent in the candidate gene approach, additional steps are required to clarify which transcripts on the list are in fact of functional importance.</p

Interseasonal RSV infections in Switzerland - rapid establishment of a clinician-led national reporting system (RSV EpiCH).

In anticipation of an interseasonal respiratory syncytial virus (RSV) epidemic, a clinician-led reporting system was rapidly established to capture RSV infections in Swiss hospitals, starting in January 2021. Here, we present details of the reporting system and first results to June 2021. An unusual epidemiology was observed with an interseasonal surge of RSV infections associated with COVID-19-related non-pharmacological interventions. These data allowed real-time adjustment of RSV prophylaxis guidelines and consequently underscore the need for and continuation of systematic nationwide RSV surveillance

Phase 3 trials of ixekizumab in moderate-to-severe plaque psoriasis

BACKGROUND Two phase 3 trials (UNCOVER-2 and UNCOVER-3) showed that at 12 weeks of treatment, ixekizumab, a monoclonal antibody against interleukin-17A, was superior to placebo and etanercept in the treatment of moderate-to-severe psoriasis. We report the 60-week data from the UNCOVER-2 and UNCOVER-3 trials, as well as 12-week and 60-week data from a third phase 3 trial, UNCOVER-1. METHODS We randomly assigned 1296 patients in the UNCOVER-1 trial, 1224 patients in the UNCOVER-2 trial, and 1346 patients in the UNCOVER-3 trial to receive subcutaneous injections of placebo (placebo group), 80 mg of ixekizumab every 2 weeks after a starting dose of 160 mg (2-wk dosing group), or 80 mg of ixekizumab every 4 weeks after a starting dose of 160 mg (4-wk dosing group). Additional cohorts in the UNCOVER-2 and UNCOVER-3 trials were randomly assigned to receive 50 mg of etanercept twice weekly. At week 12 in the UNCOVER-3 trial, the patients entered a long-term extension period during which they received 80 mg of ixekizumab every 4 weeks through week 60; at week 12 in the UNCOVER-1 and UNCOVER-2 trials, the patients who had a response to ixekizumab (defined as a static Physicians Global Assessment [sPGA] score of 0 [clear] or 1 [minimal psoriasis]) were randomly reassigned to receive placebo, 80 mg of ixekizumab every 4 weeks, or 80 mg of ixekizumab every 12 weeks through week 60. Coprimary end points were the percentage of patients who had a score on the sPGA of 0 or 1 and a 75% or greater reduction from baseline in Psoriasis Area and Severity Index (PASI 75) at week 12. RESULTS In the UNCOVER-1 trial, at week 12, the patients had better responses to ixekizumab than to placebo; in the 2-wk dosing group, 81.8% had an sPGA score of 0 or 1 and 89.1% had a PASI 75 response; in the 4-wk dosing group, the respective rates were 76.4% and 82.6%; and in the placebo group, the rates were 3.2% and 3.9% (P<0.001 for all comparisons of ixekizumab with placebo). In the UNCOVER-1 and UNCOVER-2 trials, among the patients who were randomly reassigned at week 12 to receive 80 mg of ixekizumab every 4 weeks, 80 mg of ixekizumab every 12 weeks, or placebo, an sPGA score of 0 or 1 was maintained by 73.8%, 39.0%, and 7.0% of the patients, respectively. Patients in the UNCOVER-3 trial received continuous treatment of ixekizumab from weeks 0 through 60, and at week 60, at least 73% had an sPGA score of 0 or 1 and at least 80% had a PASI 75 response. Adverse events reported during ixekizumab use included neutropenia, candidal infections, and inflammatory bowel disease. CONCLUSIONS In three phase 3 trials involving patients with psoriasis, ixekizumab was effective through 60 weeks of treatment. As with any treatment, the benefits need to be weighed against the risks of adverse events. The efficacy and safety of ixekizumab beyond 60 weeks of treatment are not yet known

Hochleistungswerkzeuge für die Blechumformung mittels Laserstrahlschmelzen

Die Fertigung von Blechumformwerkzeugen zusammen mit der Nutzung einer konturnahen Werkzeugkühlung als eine weitere mögliche Anwendung der generativen Fertigung wurde bisher kaum betrachtet. Im Rahmen des Forschungsvorhabens „HiperFormTool“ wurde daher der Einsatz des generativen Laserstrahlschmelzens für die Blechwarmumformung, das Presshärten, genauer untersucht. Hauptziel des Projektes war die Entwicklung von innovativen Werkzeugeinsätzen zur deutlichen Verbesserung des Umformprozesses. Durch eine konturnahe Werkzeugtemperierung sollten Zykluszeiten signifikant reduziert und mittels integrierter Sensorik der Umformprozess überwacht werden. Es wurden das thermische Verhalten der Werkzeuge analysiert und verschiedene Kühlgeometrien miteinander verglichen. Auf Basis der Simulationsergebnisse und unter Berücksichtigung der fertigungstechnischen Besonderheiten und mechanischer Anforderungen an das Werkzeug wurde eine innovative, grundlegend optimierte Kühlkanalgeometrie für die ausschließlich mittels Laserstrahlschmelzen mögliche Werkzeugfertigung abgeleitet. Die Sensorik wurde bereits während der generativen Fertigung in das Werkzeug integriert und erprobt. Der Vortrag beschreibt die Entwicklungen der Werkzeugtemperierung, zeigt die Vorgehensweise bei der Sensorintegration, veranschaulicht den generativen Fertigungsanlauf und gibt einen Überblick über die erreichten Ergebnisse

Potentiale generativer Fertigung im Werkzeug- und Formenbau

Die Fertigung metallischer Komponenten mit Hilfe des Laserstrahlschmelzens ermöglicht es dem Fraunhofer IWU, seinen Industriepartnern neue Wege in der Entwicklung und Fertigung innovativer Werkzeuge aufzuzeigen. Das Laserstrahlschmelzen, ein pulverbettbasiertes generatives Fertigungsverfahren, bietet dabei zahlreiche Möglichkeiten, in Werkzeugen bestehende Funktionalitäten zu verbessern bzw. auch völlig neue Funktionalitäten zu integrieren. Dabei wird zwischen geometrischer und integrativer Funktionalisierung unterscheiden. Zur geometrischen Funktionalisierung zählen beispielsweise funktionale Hohlräume und Kanäle und auch zellulare Strukturen, welche vor allem in Werkzeugen als Temperierkanäle genutzt werden. Die Gruppe der integrativen Funktionalisierung umfasst vorranging die Integration von Sensorik/Aktorik. Im vorliegenden Beitrag werden Beispiele für die Herstellung von Werkzeugen und Formen mit entsprechender Funktionalisierung vorgestellt. Es wird auf Entwicklungstrends eingegangen und Prognosen hinsichtlich Produktivitätssteigerung und Kostensenkung vorgestellt

Hochleistungswerkzeuge für die Blechwarmumformung mittels Laserstrahlschmelzen

Im Rahmen des Projektes „HiperFormTool“ wurde untersucht, wie die Blechwarmumformung durch laserstrahlgeschmolzene Werkzeugaktivkomponenten effizienter gestaltet werden kann. Dazu wurden mittels Simulation das thermische Verhalten der Werkzeuge und der Umformprozess genauestens analysiert und verschiedene Kühlkanalgeometrien miteinander verglichen. Auf Basis der Simulationsergebnisse und unter Nutzung der geometrischen Freiheiten des generativen Laserstrahlschmelzens wurde ein innovatives, konturnahes Kühlsystem abgeleitet. Hauptziel der Forschungsarbeiten war die signifikante Verkürzung der Zykluszeit. Zusätzlich wurde ein Konzept zur Sensorintegration noch während der generativen Fertigung entwickelt und umgesetzt

Laser beam melting for added value in tooling applications: Presentation held at 1st EUROFORGE conFair 2018, The future of forging, 13th - 15th Novmeber 2018, Berlin

In the recent past, additive manufacturing (AM) has been extended to more and more fields of application and is now being used in toolmaking and, in individual cases, in the series production of components. The main driving sectors are automotive production, aerospace industry and medical technology. In particular, the additive Laser Beam Melting (LBM) makes it possible – by processing conventional metallic materials such as tool steel – to manufacture innovative tools and molds. The almost limitless freedom of design of LBM enables innovative elements and complex functional structures and thus has the potential to change toolmaking. Here a distinction can be drawn between geometric and integrative functionalization. Geometric functionalization for example, includes functional cavities and channels and also cellular structures, which are mainly used in tools as cooling channels. The group of integrative functionalization primarily includes the integration of sensors as well as actuators. Plastic injection molding, light metal die casting but also metal forming such as stamping and press hardening (hot sheet metal forming) can benefit greatly from additive manufactured tooling. The presentation provides an overview of the state of the art with application examples such as the thermal management of forming tools and shows the development path for the near future