Correlation of High-Resolution X-Ray Micro-Computed Tomography with Bioluminescence Imaging of Multiple Myeloma Growth in a Xenograft Mouse Model

Multiple myeloma (MM) is an incurable B-cell neoplasia in which progressive skeletal lesions are a characteristic feature. Earlier we established an animal model for human MM in the immune-deficient RAG2-/-γc-/- mouse, in which the growth of luciferase-transduced MM cells was visualized using noninvasive bioluminescence imaging (BLI). This model appeared well suited to study disease progression and response to therapy by identifying the location of various foci of MM tumor growth scattered throughout the skeleton and at subsequent time points the quantitative assessment of the tumor load by using BLI. We report here on the corresponding high-resolution X-ray micro-computed tomographic (micro-CT) analysis to study skeletal defects in the mice with full-blown MM. Several anatomical derangements were observed, including abnormalities in geometry and morphology, asymmetrical bone structures, decreased overall density in the remaining bone, loss of trabecular bone mass, destruction of the inner microarchitecture, as well as cortical perforations. Using the combination of BLI, micro-CT imaging, and immune-histopathological techniques, we found a high correlation between the micro-CT-identified lesions, exact tumor location, and infiltration leading to structural lesions and local bone deformation. This confirms that this animal model strongly resembles human MM and has the potential for studying the biology of MM growth and for preclinical testing of novel therapies for MM and for repair of MM-induced bone lesions

A20 critically controls microglia activation and inhibits inflammasome-dependent neuroinflammation

Microglia, the mononuclear phagocytes of the central nervous system (CNS), are important for the maintenance of CNS homeostasis, but also critically contribute to CNS pathology. Here we demonstrate that the nuclear factor kappa B (NF-kappa B) regulatory protein A20 is crucial in regulating microglia activation during CNS homeostasis and pathology. In mice, deletion of A20 in microglia increases microglial cell number and affects microglial regulation of neuronal synaptic function. Administration of a sublethal dose of lipopolysaccharide induces massive microglia activation, neuroinflammation, and lethality in mice with microgliaconfined A20 deficiency. Microglia A20 deficiency also exacerbates multiple sclerosis (MS) like disease, due to hyperactivation of the NIrp3 inflammasome leading to enhanced interleukin-113 secretion and CNS inflammation. Finally, we confirm a NIrp3 inflammasome signature and IL-1 beta expression in brain and cerebrospinal fluid from MS patients. Collectively, these data reveal a critical role for A20 in the control of microglia activation and neuroinflammation

Reorganisation ofHoxdregulatory landscapes during the evolution of a snake-like body plan

Within land vertebrate species, snakes display extreme variations in their body plan, characterized by the absence of limbs and an elongated morphology. Such a particular interpretation of the basic vertebrate body architecture has often been associated with changes in the function or regulation of Hox genes. Here, we use an interspecies comparative approach to investigate different regulatory aspects at the snake HoxD locus. We report that, unlike in other vertebrates, snake mesoderm-specific enhancers are mostly located within the HoxD cluster itself rather than outside. In addition, despite both the absence of limbs and an altered Hoxd gene regulation in external genitalia, the limb-associated bimodal HoxD chromatin structure is maintained at the snake locus. Finally, we show that snake and mouse orthologous enhancer sequences can display distinct expression specificities. These results show that vertebrate morphological evolution likely involved extensive reorganisation at Hox loci, yet within a generally conserved regulatory framework.Fundação para a Ciência e Tecnologia grant: (PTDB/BEX-BID/0899/2014); Schweizerischer Nationalfonds zur Forderung der Wissenschaftlichen Forschung grant: (310030B_138662); Claraz Foundation; Université de Genève; Instituto Federal de Educação Ciência e Tecnologia do Espírito Santo

Postpartum depression—who cares? Approaches to care via midwifery, gynaecology, paediatrics and general practice

Hintergrund Die postpartale Depression (PPD) zählt zu den häufigsten psychischen Erkrankungen in der Postpartalzeit. Unbehandelt kann sie teils folgenschwere Auswirkungen auf die Mutter-Kind-Beziehung und die Entwicklung des Kindes haben. Um mögliche negative Auswirkungen verhindern zu können, sind eine frühzeitige Diagnostik betroffener Mütter und eine professionelle Betreuung essenziell. Ziel der Arbeit Der vorliegende Artikel exploriert das Zuständigkeitsgefühl der 4 Primärversorger:innen in der Postpartalzeit: Hebammen, Gynäkolog:innen, Hausärzt:innen und Pädiater:innen, und untersucht den Umgang mit der Erkrankung sowie die Barrieren und Optimierungsmöglichkeiten in der Versorgung. Material und Methoden Die primären Versorger:innen von Frauen nach einer Geburt in Deutschland wurden in 4 voneinander unabhängigen Studien befragt. Mit Hebammen, Gynäkolog:innen und Hausärzt:innen wurden quantitative Befragungen mittels Fragebögen durchgeführt, mit Vertreter:innen des deutschen Berufsverbands der Kinder- und Jugendärzte e. V. (BVKJ) eine qualitative Telefoninterviewbefragung. Es erfolgte eine systematische vergleichende Analyse. Ergebnisse und Diskussion Hebammen und Gynäkolog:innen zeigten ein deutlich höheres Zuständigkeitsgefühl für das Erkennen und die Therapie der PPD als Hausärzt:innen und Pädiater:innen. Als zentrale Voraussetzung für eine Verbesserung der Versorgungssituation in Deutschland wurden von allen 4 Berufsgruppen eine engere interdisziplinäre Zusammenarbeit und somit ein größeres Angebot an Überweisungs- und Therapiemöglichkeiten genannt. Auch eine einheitliche Regelung der finanziellen Vergütung ist für alle Versorger ein wichtiger Aspekt.Background Postpartum depression (PPD) is one of the most common mental illnesses in the postpartum period. If left untreated, it can have serious consequences for the mother–child relationship and the development of the child. In order to prevent possible negative effects, early diagnosis of affected mothers and professional care are essential. Aim of the study This article explores the sense of responsibility of the four primary care providers in the postpartum period—midwives, gynaecologists, general practitioners and paediatricians—and examines how they deal with the disease as well as the barriers and possibilities for optimisation in care. Materials and methods The primary care providers of postpartum women in Germany were interviewed in four independent studies. Quantitative questionnaires were used to interview midwives, gynaecologists and general practitioners, and a qualitative telephone survey was conducted with representatives of the German Association of Paediatricians and Adolescents (BVKJ). A systematic comparative analysis was carried out. Results and discussion Midwives and gynaecologists showed a significantly higher sense of responsibility for the recognition and treatment of PPD than general practitioners and paediatricians. Closer interdisciplinary cooperation and thus a wider range of referral and therapy options were named by all four professional groups as a central prerequisite for improving the care situation in Germany. A uniform regulation of financial remuneration is also an important aspect for all providers

Production by Clostridium acetobutylicum ATCC 824 of CelG, a Cellulosomal Glycoside Hydrolase Belonging to Family 9

The genome sequence of Clostridium acetobutylicum ATCC 824, a noncellulolytic solvent-producing strain, predicts the production of various proteins with domains typical for cellulosomal subunits. Most of the genes coding for these proteins are grouped in a cluster similar to that found in cellulolytic clostridial species, such as Clostridium cellulovorans. CAC0916, one of the open reading frames present in the putative cellulosome gene cluster, codes for CelG, a putative endoglucanase belonging to family 9, and it was cloned and overexpressed in Escherichia coli. The overproduced CelG protein was purified by making use of its high affinity for cellulose and was characterized. The biochemical properties of the purified CelG were comparable to those of other known enzymes belonging to the same family. Expression of CelG by C. acetobutylicum grown on different substrates was studied by Western blotting by using antibodies raised against the purified E. coli-produced protein. Whereas the antibodies cross-reacted with CelG-like proteins secreted by cellobiose- or cellulose-grown C. cellulovorans cultures, CelG was not detectable in extracellular medium from C. acetobutylicum grown on cellobiose or glucose. However, notably, when lichenan-grown cultures were used, several bands corresponding to CelG or CelG-like proteins were present, and there was significantly increased extracellular endoglucanase activity

Structural basis of Cullin 2 RING E3 ligase regulation by the COP9 signalosome

13 p.-5 fig.-1 tab.Cullin-Ring E3 Ligases (CRLs) regulate a multitude of cellular pathways through specific substrate receptors. The COP9 signalosome (CSN) deactivates CRLs by removing NEDD8 from activated Cullins. Here we present structures of the neddylated and deneddylated CSN-CRL2 complexes by combining single-particle cryo-electron microscopy (cryo-EM) with chemical cross-linking mass spectrometry (XL-MS). These structures suggest a conserved mechanism of CSN activation, consisting of conformational clamping of the CRL2 substrate by CSN2/CSN4, release of the catalytic CSN5/CSN6 heterodimer and finally activation of the CSN5 deneddylation machinery. Using hydrogen-deuterium exchange (HDX)-MS we show that CRL2 activates CSN5/CSN6 in a neddylation-independent manner. The presence of NEDD8 is required to activate the CSN5 active site. Overall, by synergising cryo-EM with MS, we identify sensory regions of the CSN that mediate its stepwise activation and provide a framework for understanding the regulatory mechanism of other Cullin family members. © 2019, The Author(s).The London Interdisciplinary Biosciences Consortium (LIDo) BBSRC Doctoral Training Partnership (BB/M009513/1) supports A.M.C.L. S.V.F., E.P.M. and F.B. are funded by Cancer Research UK (C12209/A16749). C.S. acknowledges funding from the Federal Ministry for Education and Research (BMBF, ZIK programme, 03Z22HN22), the European Regional Development Funds (EFRE, ZS/2016/04/78115) and the MLU Halle-Wittenberg. C.M. and A.P. are funded by the Wellcome Trust (109854/Z/15/Z) and a King’s Health Partners R&D Challenge Fund through the MRC.Peer reviewe