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PELDOR (pulse electron-electron double resonance) is an established method to study intramolecular distances and can give evidence for conformational changes and flexibilities. However, it can also be used to study intermolecular interactions as for example oligerimization. Here, we used PELDOR to study the ‘end-to-end’ stacking of small double stranded (ds)RNAs. For this study, the dsRNA molecules were only singly labelled with the spin label TPA to avoid multi-spin effects and to measure only the intermolecular stacking interactions. It can be shown that small dsRNAs tend to assemble to rod-like structures due to π-π-interactions between the base pairs at the end of the strands. On the one hand, these interactions can influence or complicate measurements aimed at the determining of the structure and dynamics of the dsRNA molecule itself. On the other hand, it can be interesting to study such intermolecular stacking interactions in more detail, as for example their dependence on ion concentration. We quantitatively determined the stacking probability as a function of the monovalent NaCl salt and the dsRNA concentration. From this data the dissociation constant Kd was deduced and found to depend on the ratio between the NaCl salt and dsRNA concentrations. Additionally, the distances and distance distributions obtained predict a model for the stacking geometry of dsRNAs. Introducing a nucleotide overhangs at one end of the dsRNA molecule restricts the stacking to the other end, leading only to dimer formations. Introducing such an overhang at both ends of the dsRNA molecule fully suppresses stacking, as we could demonstrate by PELDOR experiments quantitatively.
Prediger, Barbara ; Polus, Stephanie ; Mathes, Tim ; Bühn, Stefanie ; Louwen, Frank ; Neugebauer, Edmund ; Pieper, Dawid
Background: The rate of caesarean sections increased in the last decades to about 30% of births in western populations. Many caesarean sections are electively planned without an urgent medical reason for mother or child. Especially in women with a foregoing caesarean section, the procedure is planned early. An early caesarean section though may harm the newborn. Our aim is to evaluate the gestational time point after the 37th gestational week (after prematurity = term) of performing an elective caesarean section with the lowest morbidity for mother and child.
Methods: This is an update of a systematic review previously carried out on behalf of the German Federal Ministry of Health. We will perform a systematic literature search in MEDLINE, EMBASE, CENTRAL and CINAHL. Our primary outcome is the rate of admissions to the neonatal intensive care unit in early versus late term neonates. We will include (quasi) randomized controlled trials and cohort studies. The studies should include pregnant women who have an elective caesarean section at term. We will screen titles and abstracts and the identified full texts of studies for eligibility. Risk of bias will be assessed with the Cochrane Risk of Bias Tool for Randomized Trials or with the Risk of Bias Tool for Non-Randomized Studies of Interventions (ROBINS-I). These tasks will be performed independently by two reviewers. Data will be extracted in beforehand piloted extraction tables. A dose-response meta-analysis will be performed.
Discussion: Our aim is to reach a higher validity in the assessment of the time point of elective caesarean sections by performing a meta-analysis to support recommendations for clinical practice. We assume to identify less randomized controlled trials but a large number of cohort studies analyzing the given question. We will discuss similarities and differences in included studies as well as methodological strengths and weaknesses.
Systematic review registration: PROSPERO CRD42017078231
Steiner, Sabine ; Willfort-Ehringer, Andrea ; Sievert, Horst ; Geist, Volker ; Lichtenberg, Michael ; Del Giudice, Costantino ; Sauguet, Antoine ; Diaz-Cartelle, Juan ; Marx, Claudia ; Ströbel, Armin ; Schult, Ingolf ; Scheinert, Dierk
Objectives: The authors sought to evaluate the performance of the Ranger paclitaxel-coated balloon versus uncoated balloon angioplasty for femoropopliteal lesions at 12 months.
Background: Drug-coated balloons (DCBs) are a promising endovascular treatment option for peripheral artery disease of the femoropopliteal segment, and each unique device requires dedicated clinical study.
Methods: The prospective, randomized RANGER SFA (Comparison of the Ranger™ Paclitaxel-Coated PTA Balloon Catheter and Uncoated PTA Balloons in Femoropopliteal Arteries) study (NCT02013193) enrolled 105 patients with symptomatic lower limb ischemia (Rutherford category 2 to 4) and stenotic lesions in the nonstented femoropopliteal segment at 10 European centers. Seventy-one patients (mean age 68 ± 8 years, n = 53 men) were enrolled in the Ranger DCB arm, and 34 patients (mean age 67 ± 9 years, n = 23 men) were assigned to the control group. Twelve-month analysis included patency, safety, and clinical outcomes and quality-of-life assessments.
Results: The DCB group had a greater primary patency rate at 12 months (Kaplan-Meier estimate 86.4% vs. 56.5%), with a significantly longer time to patency failure (log-rank p < 0.001). The estimated freedom from target lesion revascularization rate was 91.2% in the DCB group and 69.9% in the control group at 12 months, with a significantly longer time to reintervention (p = 0.010). No target limb amputations or device-related deaths occurred in either group.
Conclusions: Twelve-month results show that patency was maintained longer after Ranger DCB treatment than after conventional balloon angioplasty, and this result was associated with a low revascularization rate and good clinical outcomes.
Tunaru, Sorin ; Bonnavion, Remy ; Brandenburger, Isabell ; Preussner, Jens ; Thomas, Dominique Jeanette ; Scholich, Klaus ; Offermanns, Stefan
The long-chain fatty acid receptor FFAR1 is highly expressed in pancreatic β-cells. Synthetic FFAR1 agonists can be used as antidiabetic drugs to promote glucose-stimulated insulin secretion (GSIS). However, the physiological role of FFAR1 in β-cells remains poorly understood. Here we show that 20-HETE activates FFAR1 and promotes GSIS via FFAR1 with higher potency and efficacy than dietary fatty acids such as palmitic, linoleic, and α-linolenic acid. Murine and human β-cells produce 20-HETE, and the ω-hydroxylase-mediated formation and release of 20-HETE is strongly stimulated by glucose. Pharmacological inhibition of 20-HETE formation and blockade of FFAR1 in islets inhibits GSIS. In islets from type-2 diabetic humans and mice, glucose-stimulated 20-HETE formation and 20-HETE-dependent stimulation of GSIS are strongly reduced. We show that 20-HETE is an FFAR1 agonist, which functions as an autocrine positive feed-forward regulator of GSIS, and that a reduced glucose-induced 20-HETE formation contributes to inefficient GSIS in type-2 diabetes.
Epah, Jeremy ; Pálfi, Katalin ; Dienst, Franziska Luise ; Malacarne, Pedro Felipe ; Bremer, Rolf ; Salamon, Michael ; Kumar, Sandeep ; Jo, Hanjoong ; Schürmann, Christoph ; Brandes, Ralf
Rationale: Classic histology is the gold standard for vascular network imaging and analysis. The method however is laborious and prone to artefacts. Here, the suitability of ultramicroscopy (UM) and micro-computed tomography (CT) was studied to establish potential alternatives to histology.
Methods: The vasculature of murine organs (kidney, heart and atherosclerotic carotid arteries) was visualized using conventional 2D microscopy, 3D light sheet ultramicroscopy (UM) and micro-CT. Moreover, spheroid-based human endothelial cell vessel formation in mice was quantified. Fluorescently labeled Isolectin GS-IB4 A647 was used for in vivo labeling of vasculature for UM analysis, and analyses were performed ex vivo after sample preparation. For CT imaging, animals were perfused postmortem with radiopaque contrast agent.
Results: Using UM imaging, 3D vascular network information could be obtained in samples of animals receiving in vivo injection of the fluorescently labeled Isolectin GS-IB4. Resolution was sufficient to measure single endothelial cell integration into capillaries in the spheroid-based matrigel plug assay. Because of the selective staining of the endothelium, imaging of larger vessels yielded less favorable results. Using micro-CT or even nano-CT, imaging of capillaries was impossible due to insufficient X-ray absorption and thus insufficient signal-to-noise ratio. Identification of lumen in murine arteries using micro-CT was in contrast superior to UM.
Conclusion: UM and micro-CT are two complementary techniques. Whereas UM is ideal for imaging and especially quantifying capillary networks and arterioles, larger vascular structures are easier and faster to quantify and visualize using micro-CT. 3D information of both techniques is superior to 2D histology. UM and micro-CT together may open a new field of clinical pathology diagnosis.
Tran, Bach Xuan ; Pham, Thang Van ; Ha, Giang Hai ; Ngo, Anh Toan ; Nguyen, Long Hoang ; Vu, Thuc Thi Minh ; Do, Ha Ngoc ; Nguyen, Vu ; Nguyen, Anh Tuan Le ; Tran, Tung Thanh ; Truong, Nu Thi ; Hoang, Vuong Quan ; Ho, Tung Manh ; Dam, Nhue Van ; Vuong, Thu Trang ; Nguyen, Hung Quang ; Le, Huong Thi ; Do, Hoa Thi ; Moir, Mackenzie ; Shimpuku, Yoko ; Dhimal, Meghnath ; Arya, Shalini Subash ; Nguyen, Tu Huu ; Bhattarai, Suraj ; Latkin, Carl A. ; Ho, Cyrus S. H. ; Ho, Roger C. M.
Child maltreatment remains a major health threat globally that requires the understanding of socioeconomic and cultural contexts to craft effective interventions. However, little is known about research agendas globally and the development of knowledge-producing networks in this field of study. This study aims to explore the bibliometric overview on child maltreatment publications to understand their growth from 1916 to 2018. Data from the Web of Science Core Collection were collected in May 2018. Only research articles and reviews written in the English language were included, with no restrictions by publication date. We analyzed publication years, number of papers, journals, authors, keywords and countries, and presented the countries collaboration and co-occurrence keywords analysis. From 1916 to 2018, 47,090 papers (53.0% in 2010–2018) were published in 9442 journals. Child Abuse & Neglect (2576 papers; 5.5%); Children and Youth Services Review (1130 papers; 2.4%) and Pediatrics (793 papers, 1.7%) published the most papers. The most common research areas were Psychology (16,049 papers, 34.1%), Family Studies (8225 papers, 17.5%), and Social Work (7367 papers, 15.6%). Among 192 countries with research publications, the most prolific countries were the United States (26,367 papers), England (4676 papers), Canada (3282 papers) and Australia (2664 papers). We identified 17 authors who had more than 60 scientific items. The most cited papers (with at least 600 citations) were published in 29 journals, headed by the Journal of the American Medical Association (JAMA) (7 papers) and the Lancet (5 papers). This overview of global research in child maltreatment indicated an increasing trend in this topic, with the world’s leading centers located in the Western countries led by the United States. We called for interdisciplinary research approaches to evaluating and intervening on child maltreatment, with a focus on low-middle income countries (LMICs) settings and specific contexts.
Despite being generally distributed and common on Cuba, Hispaniola, and in south Florida, species of Pyrgus Hübner (Lepidoptera: Hesperiidae), commonly known as checkered skippers, are very poorly known from The Bahamas. Previous records indicated the presence only of Pyrgus oileus (Linnaeus, 1767), just from Great Inagua Island, although its status on that island remains unclear. Herein we document P. oileus for the first time from Grand Bahama Island, suggesting an independent dispersal of this species to the northern Bahamas from south Florida. Furthermore, we document Pyrgus albescens Plötz, 1884 from Grand Bahama and Abaco islands, representing the first Caribbean records for this rapidly dispersing species. We suggest that both P. oileus and P. albescens arrived on Grand Bahama sometime between 2010 and 2014, most likely from south Florida, and that P. albescens has subsequently dispersed to Abaco. Careful study of Pyrgus species in The Bahamas is needed to document future colonization events.
Hasson, Uri ; Iacovacci, Jacopo ; Davis, Ben ; Flanagan, Ryan ; Tagliazucchi, Enzo ; Laufs, Helmut ; Lacasa, Lucas
We explore a combinatorial framework which efficiently quantifies the asymmetries between minima and maxima in local fluctuations of time series. We first showcase its performance by applying it to a battery of synthetic cases. We find rigorous results on some canonical dynamical models (stochastic processes with and without correlations, chaotic processes) complemented by extensive numerical simulations for a range of processes which indicate that the methodology correctly distinguishes different complex dynamics and outperforms state of the art metrics in several cases. Subsequently, we apply this methodology to real-world problems emerging across several disciplines including cases in neurobiology, finance and climate science. We conclude that differences between the statistics of local maxima and local minima in time series are highly informative of the complex underlying dynamics and a graph-theoretic extraction procedure allows to use these features for statistical learning purposes.
The prediction of protein–ligand interactions and their corresponding binding free energy is a challenging task in structure-based drug design and related applications. Docking and scoring is broadly used to propose the binding mode and underlying interactions as well as to provide a measure for ligand affinity or differentiate between active and inactive ligands. Various studies have revealed that most docking software packages reliably predict the binding mode, although scoring remains a challenge. Here, a diverse benchmark data set of 99 matched molecular pairs (3D-MMPs) with experimentally determined X-ray structures and corresponding binding affinities is introduced. This data set was used to study the predictive power of 13 commonly used scoring functions to demonstrate the applicability of the 3D-MMP data set as a valuable tool for benchmarking scoring functions.
Brinker, Titus Josef ; Alfitian, Jonas ; Seeger, Werner ; Groneberg, Jan David Alexander ; Kalle, Christof von ; Enk, Alexander ; Herth, Felix ; Kreuter, Michael ; Bauer, Claudia M. ; Gatzka, Martina ; Suhre, Janina Leonie
The Education Against Tobacco (EAT) network delivers smoking prevention advice in secondary schools, typically using the mirroring approach (i.e., a "selfie" altered with a face-aging app and shared with a class). In November 2017, however, the German assembly of EAT opted to expand its remit to include nursing students. To assess the transferability of the existing approach, we implemented it with the self-developed face-aging app "Smokerface" (=mixed − methods approach) in six nursing schools. Anonymous questionnaires were used to assess the perceptions of 197 students (age 18–40 years; 83.8% female; 26.4% smokers; 23.3% daily smokers) collecting qualitative and quantitative data for our cross-sectional study. Most students perceived the intervention to be fun (73.3%), but a minority disagreed that their own animated selfie (25.9%) or the reaction of their peers (29.5%) had motivated them to stop smoking. The impact on motivation not to smoke was considerably lower than experienced with seventh graders (63.2% vs. 42.0%; notably, more smokers also disagreed (45.1%) than agreed (23.5%) with this statement. Agreement rates on the motivation not to smoke item were higher in females than in males and in year 2–3 than in year 1 students. Potential improvements included greater focus on pathology (29%) and discussing external factors (26%). Overall, the intervention seemed to be appealing for nursing students
Hommers, Leif ; Richter, Jan ; Yang, Yunbo ; Raab, Annette ; Baumann, Christian ; Lang, Konstantin ; Schiele, Miriam A. ; Weber, Heike ; Wittmann, André ; Wolf, Christiane ; Alpers, Georg W. ; Arolt, Volker ; Domschke, Katharina ; Fehm, Lydia ; Fydrich, Thomas ; Gerlach, Alexander ; Gloster, Andrew T. ; Hamm, Alfons ; Helbig-Lang, Sylvia ; Kircher, Tilo ; Lang, Thomas ; Pané-Farré, Christiane A. ; Pauli, Paul ; Pfleiderer, Bettina ; Reif, Andreas ; Romanos, Marcel ; Straube, Benjamin ; Ströhle, Andreas ; Wittchen, Hans-Ulrich ; Frantz, Stefan ; Ertl, Georg ; Lohse, Martin ; Lüken, Ulrike ; Deckert, Jürgen
Increased sympathetic noradrenergic signaling is crucially involved in fear and anxiety as defensive states. MicroRNAs regulate dynamic gene expression during synaptic plasticity and genetic variation of microRNAs modulating noradrenaline transporter gene (SLC6A2) expression may thus lead to altered central and peripheral processing of fear and anxiety. In silico prediction of microRNA regulation of SLC6A2 was confirmed by luciferase reporter assays and identified hsa-miR-579-3p as a regulating microRNA. The minor (T)-allele of rs2910931 (MAFcases = 0.431, MAFcontrols = 0.368) upstream of MIR579 was associated with panic disorder in patients (pallelic = 0.004, ncases = 506, ncontrols = 506) and with higher trait anxiety in healthy individuals (pASI = 0.029, pACQ = 0.047, n = 3112). Compared to the major (A)-allele, increased promoter activity was observed in luciferase reporter assays in vitro suggesting more effective MIR579 expression and SLC6A2 repression in vivo (p = 0.041). Healthy individuals carrying at least one (T)-allele showed a brain activation pattern suggesting increased defensive responding and sympathetic noradrenergic activation in midbrain and limbic areas during the extinction of conditioned fear. Panic disorder patients carrying two (T)-alleles showed elevated heart rates in an anxiety-provoking behavioral avoidance test (F(2, 270) = 5.47, p = 0.005). Fine-tuning of noradrenaline homeostasis by a MIR579 genetic variation modulated central and peripheral sympathetic noradrenergic activation during fear processing and anxiety. This study opens new perspectives on the role of microRNAs in the etiopathogenesis of anxiety disorders, particularly their cardiovascular symptoms and comorbidities.
Petrone, Pierpaolo ; Pucci, Piero ; Vergara, Alessandro ; Amoresano, Angela ; Birolo, Leila ; Pane, Francesca ; Sirano, Francesco ; Niola, Massimo ; Buccelli, Claudio ; Graziano, Vincenzo
In AD 79 the town of Herculaneum was suddenly hit and overwhelmed by volcanic ash-avalanches that killed all its remaining residents, as also occurred in Pompeii and other settlements as far as 20 kilometers from Vesuvius. New investigations on the victims' skeletons unearthed from the ash deposit filling 12 waterfront chambers have now revealed widespread preservation of atypical red and black mineral residues encrusting the bones, which also impregnate the ash filling the intracranial cavity and the ash-bed encasing the skeletons. Here we show the unique detection of large amounts of iron and iron oxides from such residues, as revealed by inductively coupled plasma mass spectrometry and Raman microspectroscopy, thought to be the final products of heme iron upon thermal decomposition. The extraordinarily rare preservation of significant putative evidence of hemoprotein thermal degradation from the eruption victims strongly suggests the rapid vaporization of body fluids and soft tissues of people at death due to exposure to extreme heat.
Kringel, Dario ; Lippmann, Christian ; Parnham, Michael J. ; Kalso, Eija ; Ultsch, Alfred ; Lötsch, Jörn
Background: Human genetic research has implicated functional variants of more than one hundred genes in the modulation of persisting pain. Artificial intelligence and machine‐learning techniques may combine this knowledge with results of genetic research gathered in any context, which permits the identification of the key biological processes involved in chronic sensitization to pain.
Methods: Based on published evidence, a set of 110 genes carrying variants reported to be associated with modulation of the clinical phenotype of persisting pain in eight different clinical settings was submitted to unsupervised machine‐learning aimed at functional clustering. Subsequently, a mathematically supported subset of genes, comprising those most consistently involved in persisting pain, was analysed by means of computational functional genomics in the Gene Ontology knowledgebase.
Results: Clustering of genes with evidence for a modulation of persisting pain elucidated a functionally heterogeneous set. The situation cleared when the focus was narrowed to a genetic modulation consistently observed throughout several clinical settings. On this basis, two groups of biological processes, the immune system and nitric oxide signalling, emerged as major players in sensitization to persisting pain, which is biologically highly plausible and in agreement with other lines of pain research.
Conclusions: The present computational functional genomics‐based approach provided a computational systems‐biology perspective on chronic sensitization to pain. Human genetic control of persisting pain points to the immune system as a source of potential future targets for drugs directed against persisting pain. Contemporary machine‐learned methods provide innovative approaches to knowledge discovery from previous evidence.
Significance: We show that knowledge discovery in genetic databases and contemporary machine‐learned techniques can identify relevant biological processes involved in Persitent pain.
Kwankamol Limsopatham ; Hall, Martin J. R. ; Zehner, Richard ; Zajac, Barbara Karolina ; Verhoff, Marcel A. ; Narin Sontigun ; Kom Sukontason ; Kabkaew L. Sukontason ; Amendt, Jens
The bluebottle blow fly Calliphora vicina is a common species distributed throughout Europe that can play an important role as forensic evidence in crime investigations. Developmental rates of C. vicina from distinct populations from Germany and England were compared under different temperature regimes to explore the use of growth data from different geographical regions for local case work. Wing morphometrics and molecular analysis between these populations were also studied as indicators for biological differences. One colony each of German and English C. vicina were cultured at the Institute of Legal Medicine in Frankfurt, Germany. Three different temperature regimes were applied, two constant (16°C & 25°C) and one variable (17–26°C, room temperature = RT). At seven time points (600, 850, 1200, 1450, 1800, 2050, and 2400 accumulated degree hours), larval lengths were measured; additionally, the durations of the post feeding stage and intrapuparial metamorphosis were recorded. For the morphometric and molecular study, 184 females and 133 males from each C. vicina population (Germany n = 3, England n = 4) were sampled. Right wings were measured based on 19 landmarks and analyzed using canonical variates analysis and discriminant function analysis. DNA was isolated from three legs per specimen (n = 61) using 5% chelex. A 784 bp long fragment of the mitochondrial cytochrome b gene was sequenced; sequences were aligned and phylogenetically analyzed. Similar larval growth rates of C. vicina were found from different geographic populations at different temperatures during the major part of development. Nevertheless, because minor differences were found a wider range of temperatures and sampling more time points should be analyzed to obtain more information relevant for forensic case work. Wing shape variation showed a difference between the German and English populations (P<0.0001). However, separation between the seven German and English populations at the smaller geographic scale remained ambiguous. Molecular phylogenetic analysis by maximum likelihood method could not unambiguously separate the different geographic populations at a national (Germany vs England) or local level.
Anomiopus cirulito Cano n. sp., from the tropical forest of the Mayan Biosphere Reserve, Petén, Guatemala is described. This is the northernmost known species of the genus Anomiopus Westwood (Coleoptera: Scarabaeidae: Scarabaeinae) and is related to the Costa Rican and Panamanian Anomiopus panamensis (Paulian).
Classifying and mapping landscapes are tools to simplify complex systems into the discreet subsets widely used in landscape management. In 1999, the Queensland Government adopted a Regional Ecosystems approach as a state-wide landscape classification scheme. For the Cape York Peninsula bioregion in north-eastern Australia, Regional Ecosystems (RE) were initially recognised based on a pre-existing vegetation map and classification for the bioregion. The classification had been developed using expert-techniques based on extensive field plot data. Here, we use numerical analyses to classify the field plot data and identify savanna plant communities associated with two widespread landform groups in the bioregion (the old loamy and sandy plains (land zone 5) and the hills and lowlands on igneous rocks (land zone 12). Communities were identified at the plant association level, using species importance values calculated from foliage cover and vegetation height at each plot. We developed a descriptive-framework for each community using statistically based characterising species and biophysical attributes. We recognise 57 communities compared with 110 that had been previously identified using expert-techniques. This classification is used to recommend refined Regional Ecosystems under the government’s regulations. The descriptive-framework supported consistent descriptions of communities and assignment of new sites to the classification. We conclude that incorporating quantitative methods in classifying and describing plant communities will improve the robustness and defensibility of Regional Ecosystems and their use in landscape management across Queensland.
Constantino, Luis Miguel ; Cadena-Castañeda, Oscar J. ; Cardona Granda, Juan Manuel ; Benavides Machado, Pablo ; Góngora Botero, Carmenza
A remarkable new species of Poecilocloeus (Orthoptera: Acrididae: Proctolabinae) found damaging coffee plantations in the Western Andes of Colombia is described and named P. coffeaphilus n. sp. This new species is part of a distinct and colorful group of Proctolabinae grasshoppers, with most species found at low altitudes in the rainforest of the Amazon basin. In contrast, the new species is found at elevations of 1600 to 1800 m in the canopy of dense cloud forests, in the southwestern part of the department of Antioquia (Western Cordillera, Colombian Andes). Information about the natural history, behavior, natural enemies and control strategies in coffee plantations is given for this new species of masked grasshopper. A key to the Neotropical species of the fruticolus species group is presented.
Zomot, Elia ; Yardeni, Eliane Hadas ; Vargiu, Attilio Vittorio ; Tam, Heng Keat ; Malloci, Giuliano ; Ramaswamy, Venkata Krishnan ; Perach, Michal ; Ruggerone, Paolo ; Pos, Klaas Martinus ; Bibi, Eitan
Secondary multidrug (Mdr) transporters utilize ion concentration gradients to actively remove antibiotics and other toxic compounds from cells. The model Mdr transporter MdfA from Escherichia coli exchanges dissimilar drugs for protons. The transporter should open at the cytoplasmic side to enable access of drugs into the Mdr recognition pocket. Here we show that the cytoplasmic rim around the Mdr recognition pocket represents a previously overlooked important regulatory determinant in MdfA. We demonstrate that increasing the positive charge of the electrically asymmetric rim dramatically inhibits MdfA activity and sometimes even leads to influx of planar, positively charged compounds, resulting in drug sensitivity. Our results suggest that unlike the mutants with the electrically modified rim, the membrane-embedded wild-type MdfA exhibits a significant probability of an inward-closed conformation, which is further increased by drug binding. Since MdfA binds drugs from its inward-facing environment, these results are intriguing and raise the possibility that the transporter has a sensitive, drug-induced conformational switch, which favors an inward-closed state.
A new species of leaf insect from the celebicum species group, Phyllium (Phyllium) yapicum Cumming and Teemsma, new species (Phasmida: Phylliidae), is described from a female specimen from the California Academy of Sciences collection, United States. This new species is the first recorded species of Phylliidae from the country of Micronesia and represents a notable range expansion for the family. With Phyllium (Phyllium) yapicum Cumming and Teemsma, new species, currently only known from a female holotype; a key to females is included for the celebicum species group.
Weigel, Robert ; Irl, Severin ; Treydte, Kerstin ; Beierkuhnlein, Carl ; Berels, Johanna ; Field, Richard ; Miranda, José Carlos ; Steinbauer, Alana ; Steinbauer, Manuel ; Jentsch, Anke
Climatic seasonality drives ecosystem processes (e.g. productivity) and influences plant species distribution. However, it is poorly understood how different aspects of seasonality (especially regarding temperature and precipitation) affect growth continuity of trees in climates with low seasonality because seasonality is often only crudely measured. On islands, exceptionally wide elevational species distribution ranges allow the use of tree rings to identify how growth continuity and climate–growth relationships change with elevation. Here, we present a novel dendroecological method to measure stem growth continuity based on annual density fluctuations (ADFs) in tree rings of Pinus canariensis to indicate low climatic seasonality. The species ranges from 300 to >2000 m a.s.l. on the trade wind-influenced island of La Palma (Canary Islands), where we measured three decades of tree-ring data of 100 individuals distributed over 10 sites along the entire elevational range. The successfully implemented ADF approach revealed a major shift of stem growth continuity across the elevational gradient. In a remarkably clear pattern, stem growth continuity (percentage of ADFs) showed a hump-shaped relationship with elevation reaching a maximum at around 1000 m a.s.l. Low- to mid-elevation tree growth was positively correlated with the Palmer Drought Severity Index (PDSI; indicating aridity) and sea surface temperature (indicating trade wind-influenced moderation of water supply), while high-elevation tree growth was positively correlated with winter temperature (indicating a cold-induced dormancy period). We conclude that ADFs are a useful method to measure stem growth continuity in low-seasonality climates. Growth of P. canariensis on the Canary Islands is more frequently interrupted by winter cold at high elevations and by summer drought at low elevations than in the trade wind-influenced mid elevations, where growth sometimes continues throughout the year. Climate change-associated alterations in trade wind cloud formation might cause non-analogue growth limitations for many unique island species.
Snodgrass, Ryan G. ; Zezina, Ekaterina ; Namgaladze, Dmitry ; Gupta, Sahil ; Angioni, Carlo Federico ; Geisslinger, Gerd ; Lütjohann, Dieter ; Brüne, Bernhard
Arachidonate 15-lipoxygenase (ALOX15) and arachidonate 15-lipoxygenase, type B (ALOX15B) catalyze the dioxygenation of polyunsaturated fatty acids and are upregulated in human alternatively activated macrophages (AAMs) induced by Th2 cytokine interleukin-4 (IL-4) and/or interleukin-13. Known primarily for roles in bioactive lipid mediator synthesis, 15-lipoxygenases (15-LOXs) have been implicated in various macrophage functions including efferocytosis and ferroptosis. Using a combination of inhibitors and siRNAs to suppress 15-LOX isoforms, we studied the role of 15-LOXs in cellular cholesterol homeostasis and immune function in naïve and AAMs. Silencing or inhibiting the 15-LOX isoforms impaired sterol regulatory element binding protein (SREBP)-2 signaling by inhibiting SREBP-2 processing into mature transcription factor and reduced SREBP-2 binding to sterol regulatory elements and subsequent target gene expression. Silencing ALOX15B reduced cellular cholesterol and the cholesterol intermediates desmosterol, lanosterol, 24,25-dihydrolanosterol, and lathosterol as well as oxysterols in IL-4-stimulated macrophages. In addition, attenuating both 15-LOX isoforms did not generally affect IL-4 gene expression but rather uniquely impacted IL-4-induced CCL17 production in an SREBP-2-dependent manner resulting in reduced T cell migration to macrophage conditioned media. In conclusion, we identified a novel role for ALOX15B, and to a lesser extent ALOX15, in cholesterol homeostasis and CCL17 production in human macrophages.
Of the circa 25 species of butterfly species occurring in the Cabo Verde Archipelago, only one species is endemic: Chilades evorae (Lycaenidae). The species was reared by the authors and colour photographs of the early stages of this species are presented for the first time. The host-plants on the islands of Santo Antão, São Vicente, and Santa Luzia are illustrated with photographs. The butterfly is reported from Raso for the first time, together with a potential host-plant. A tachinid fly species Cadurciella sp. parasitising C. evorae is also reported.
A point mutation in the Ncr1 signal peptide impairs the development of innate lymphoid cell subsets
(2018)
Almeida, Francisca F. ; Tognarelli, Sara ; Marçais, Antoine ; Kueh, Andrew J. ; Friede, Miriam Elisabeth ; Liao, Yang ; Willis, Simon N. ; Luong, Kylie ; Faure, Fabrice ; Mercier, Francois E. ; Galluso, Justine ; Firth, Matthew ; Narni-Mancinelli, Emilie ; Rais, Bushra ; Scadden, David T. ; Spallotta, Francesco ; Weil, Sandra ; Giannattasio, Ariane ; Kalensee, Franziska ; Zöller, Tobias ; Huntington, Nicholas D. ; Schleicher, Ulrike ; Geburtig-Chiocchetti, Andreas ; Ugolini, Sophie ; Herold, Marco ; Shi, Wei ; Koch, Joachim ; Steinle, Alexander ; Vivier, Eric ; Walzer, Thierry ; Belz, Gabrielle T. ; Ullrich, Evelyn
NKp46 (CD335) is a surface receptor shared by both human and mouse natural killer (NK) cells and innate lymphoid cells (ILCs) that transduces activating signals necessary to eliminate virus-infected cells and tumors. Here, we describe a spontaneous point mutation of cysteine to arginine (C14R) in the signal peptide of the NKp46 protein in congenic Ly5.1 mice and the newly generated NCRB6C14R strain. Ly5.1C14R NK cells expressed similar levels of Ncr1 mRNA as C57BL/6, but showed impaired surface NKp46 and reduced ability to control melanoma tumors in vivo. Expression of the mutant NKp46C14R in 293T cells showed that NKp46 protein trafficking to the cell surface was compromised. Although Ly5.1C14R mice had normal number of NK cells, they showed an increased number of early maturation stage NK cells. CD49a+ILC1s were also increased but these cells lacked the expression of TRAIL. ILC3s that expressed NKp46 were not detectable and were not apparent when examined by T-bet expression. Thus, the C14R mutation reveals that NKp46 is important for NK cell and ILC differentiation, maturation and function.
Rooijen, Mathilde van ; Jairam, An ; Tollens, Tim ; Jørgensen, Lars Nannestad ; Vries Reilingh, Tammo S. de ; Piessen, Guillaume ; Köckerling, Ferdinand ; Miserez, Marc ; Windsor, Alastair C. J. ; Berrevoet, Frederik ; Fortelny, René H. ; Dousset, Bertrand ; Woeste, Guido ; Westreenen, Henderik Leendert van ; Gossetti, Francesco ; Lange, Johan F. ; Tetteroo, Geert W. M. ; Koch, Andreas ; Kroese, Leonard F. ; Jeekel, Johannes
Background: Incisional heia is a frequent complication of midline laparotomy. The use of mesh in hernia repair has been reported to lead to fewer recurrences compared to primary repair. However, in Ventral Hernia Working Group (VHWG) Grade 3 hernia patients, whose hernia is potentially contaminated, synthetic mesh is prone to infection. There is a strong preference for resorbable biological mesh in contaminated fields, since it is more able to resist infection, and because it is fully resorbed, the chance of a foreign body reaction is reduced. However, when not crosslinked, biological resorbable mesh products tend to degrade too quickly to facilitate native cellular ingrowth. Phasix™ Mesh is a biosynthetic mesh with both the biocompatibility and resorbability of a biological mesh and the mechanical strength of a synthetic mesh. This multi-center single-arm study aims to collect data on safety and performance of Phasix™ Mesh in Grade 3 hernia patients.
Methods: A total of 85 VHWG Grade 3 hernia patients will be treated with Phasix™ Mesh in 15 sites across Europe. The primary outcome is Surgical Site Occurrence (SSO) including hematoma, seroma, infection, dehiscence and fistula formation (requiring intervention) through 3 months. Secondary outcomes include recurrence, infection and quality of life related outcomes after 24 months. Follow-up visits will be at drain removal (if drains were not placed, then on discharge or staple removal instead) and in the 1st, 3rd, 6th, 12th, 18th and 24th month after surgery.
Conclusion: Based on evidence from this clinical study Depending on the results this clinical study will yield, Phasix™ Mesh may become a preferred treatment option in VHWG Grade 3 patients.
Trial registration: The trial was registered on March 25, 2016 on clinicaltrials.gov: NCT02720042.
Cabo Verde is a unique biogeographical region, where by mixing temperate and tropical characteristics an unusually high number of endemic species are reported. Cephalopods are central pieces of trophic networks worldwide, interacting as predator/ prey and competing with fish for ecological niches. We aimed to assess how the topography, prey availability, and predatory pressure of the Desertas Islands shaped the behaviour and ecology of the existing Octopus vulgaris population. Visual census (both underwater and on tidal rock pools) were performed on Santa Luzia Island (20 days) and Raso Islet (eight days). Octopus vulgaris individuals were found only in intertidal areas, during low tide, and mean population morphometry averaged 35.6 ± 10.4 cm (total length) and 175.0 ± 53 g (wet weight). The markedly reduced size of O. vulgaris, only partly explainable by Bergmann’s rule, and exclusion from subtidal areas, appears to have been mainly driven by severe predatory pressure and strong inter-specific competition for limited habitat niches. The induced behavioural and morphometric alterations may be the product of developmental plasticity, or have arisen from deeper genetic alterations, which would portray a potential speciation phenomenon of octopus’ populations residing on Cabo Verde’s Desertas Islands.
Acetaminophen [paracetamol, N-acetyl-p-aminophenol (APAP)]-induced acute liver injury (ALI) not only remains a persistent clinical challenge but likewise stands out as well-characterized paradigmatic model of drug-induced liver damage. APAP intoxication associates with robust hepatic necroinflammation the role of which remains elusive with pathogenic but also pro-regenerative/-resolving functions being ascribed to leukocyte activation. Here, we shine a light on and put forward a unique role of the interleukin (IL)-1 family member IL-18 in experimental APAP-induced ALI. Indeed, amelioration of disease as previously observed in IL-18-deficient mice was further substantiated herein by application of the IL-18 opponent IL-18-binding protein (IL-18BPd:Fc) to wild-type mice. Data altogether emphasize crucial pathological action of this cytokine in APAP toxicity. Adding recombinant IL-22 to IL-18BPd:Fc further enhanced protection from liver injury. In contrast to IL-18, the role of prototypic pro-inflammatory IL-1 and tumor necrosis factor-α is controversially discussed with lack of effects or even protective action being repeatedly reported. A prominent detrimental function for IL-18 in APAP-induced ALI as proposed herein should relate to its pivotal role for hepatic expression of interferon-γ and Fas ligand, both of which aggravate APAP toxicity. As IL-18 serum levels increase in patients after APAP overdosing, targeting IL-18 may evolve as novel therapeutic option in those hard-to-treat patients where standard therapy with N-acetylcysteine is unsuccessful. Being a paradigmatic experimental model of ALI, current knowledge on ill-fated properties of IL-18 in APAP intoxication likewise emphasizes the potential of this cytokine to serve as therapeutic target in other entities of inflammatory liver diseases.
Kaese, Thomas ; Trageser, Timo ; Budy, Hendrik ; Bolte, Michael ; Lerner, Hans-Wolfram ; Wagner, Matthias
Organoboranes are among the most versatile and widely used reagents in synthetic chemistry. A significant further expansion of their application spectrum would be achievable if boron-containing reactive intermediates capable of inserting into C–H bonds or performing nucleophilic substitution reactions were readily available. However, current progress in the field is still hampered by a lack of universal design concepts and mechanistic understanding. Herein we report that the doubly arylene-bridged diborane(6) 1H2 and its B[double bond, length as m-dash]B-bonded formal deprotonation product Li2[1] can activate the particularly inert C(sp3)–H bonds of added H3CLi and H3CCl, respectively. The first case involves the attack of [H3C]− on a Lewis-acidic boron center, whereas the second case follows a polarity-inverted pathway with nucleophilic attack of the B[double bond, length as m-dash]B double bond on H3CCl. Mechanistic details were elucidated by means of deuterium-labeled reagents, a radical clock, α,ω-dihaloalkane substrates, the experimental identification of key intermediates, and quantum-chemical calculations. It turned out that both systems, H3CLi/1H2 and H3CCl/Li2[1], ultimately funnel into the same reaction pathway, which likely proceeds past a borylene-type intermediate and requires the cooperative interaction of both boron atoms.
Mishra, Bagdevi ; Gupta, Deepak Kumar ; Pfenninger, Markus ; Hickler, Thomas ; Langer, Ewald ; Nam, Bora ; Paule, Juraj ; Sharma, Rahul ; Ulaszewski, Bartosz ; Warmbier, Joanna ; Burczyk, Jarosław ; Thines, Marco
Background: The European beech is arguably the most important climax broad-leaved tree species in Central Europe, widely planted for its valuable wood. Here, we report the 542 Mb draft genome sequence of an up to 300-year-old individual (Bhaga) from an undisturbed stand in the Kellerwald-Edersee National Park in central Germany.
Findings: Using a hybrid assembly approach, Illumina reads with short- and long-insert libraries, coupled with long Pacific Biosciences reads, we obtained an assembled genome size of 542 Mb, in line with flow cytometric genome size estimation. The largest scaffold was of 1.15 Mb, the N50 length was 145 kb, and the L50 count was 983. The assembly contained 0.12% of Ns. A Benchmarking with Universal Single-Copy Orthologs (BUSCO) analysis retrieved 94% complete BUSCO genes, well in the range of other high-quality draft genomes of trees. A total of 62,012 protein-coding genes were predicted, assisted by transcriptome sequencing. In addition, we are reporting an efficient method for extracting high-molecular-weight DNA from dormant buds, by which contamination by environmental bacteria and fungi was kept at a minimum.
Conclusions: The assembled genome will be a valuable resource and reference for future population genomics studies on the evolution and past climate change adaptation of beech and will be helpful for identifying genes, e.g., involved in drought tolerance, in order to select and breed individuals to adapt forestry to climate change in Europe. A continuously updated genome browser and download page can be accessed from beechgenome.net, which will include future genome versions of the reference individual Bhaga, as new sequencing approaches develop.
Engel, Andreas ; Bönisch, Harald ; Ostermöller, Jennifer ; Chipperfield, Martyn P. ; Dhomse, Sandip ; Jöckel, Patrick
Chlorine and bromine atoms lead to catalytic depletion of ozone in the stratosphere. Therefore the use and production of ozone-depleting substances (ODSs) containing chlorine and bromine is regulated by the Montreal Protocol to protect the ozone layer. Equivalent effective stratospheric chlorine (EESC) has been adopted as an appropriate metric to describe the combined effects of chlorine and bromine released from halocarbons on stratospheric ozone. Here we revisit the concept of calculating EESC. We derive a refined formulation of EESC based on an advanced concept of ODS propagation into the stratosphere and reactive halogen release. A new transit time distribution is introduced in which the age spectrum for an inert tracer is weighted with the release function for inorganic halogen from the source gases. This distribution is termed the release time distribution. We show that a much better agreement with inorganic halogen loading from the chemistry transport model TOMCAT is achieved compared with using the current formulation. The refined formulation shows EESC levels in the year 1980 for the mid-latitude lower stratosphere, which are significantly lower than previously calculated. The year 1980 is commonly used as a benchmark to which EESC must return in order to reach significant progress towards halogen and ozone recovery. Assuming that – under otherwise unchanged conditions – the EESC value must return to the same level in order for ozone to fully recover, we show that it will take more than 10 years longer than estimated in this region of the stratosphere with the current method for calculation of EESC. We also present a range of sensitivity studies to investigate the effect of changes and uncertainties in the fractional release factors and in the assumptions on the shape of the release time distributions. We further discuss the value of EESC as a proxy for future evolution of inorganic halogen loading under changing atmospheric dynamics using simulations from the EMAC model. We show that while the expected changes in stratospheric transport lead to significant differences between EESC and modelled inorganic halogen loading at constant mean age, EESC is a reasonable proxy for modelled inorganic halogen on a constant pressure level.
Neurons collect their inputs from other neurons by sending out arborized dendritic structures. However, the relationship between the shape of dendrites and the precise organization of synaptic inputs in the neural tissue remains unclear. Inputs could be distributed in tight clusters, entirely randomly or else in a regular grid-like manner. Here, we analyze dendritic branching structures using a regularity index R, based on average nearest neighbor distances between branch and termination points, characterizing their spatial distribution. We find that the distributions of these points depend strongly on cell types, indicating possible fundamental differences in synaptic input organization. Moreover, R is independent of cell size and we find that it is only weakly correlated with other branching statistics, suggesting that it might reflect features of dendritic morphology that are not captured by commonly studied branching statistics. We then use morphological models based on optimal wiring principles to study the relation between input distributions and dendritic branching structures. Using our models, we find that branch point distributions correlate more closely with the input distributions while termination points in dendrites are generally spread out more randomly with a close to uniform distribution. We validate these model predictions with connectome data. Finally, we find that in spatial input distributions with increasing regularity, characteristic scaling relationships between branching features are altered significantly. In summary, we conclude that local statistics of input distributions and dendrite morphology depend on each other leading to potentially cell type specific branching features.
An unusual eye malformation observed in Trichiotinus rufobrunneus (Casey) (Coleoptera: Scarabaeidae: Cetoniinae: Trichiini), is described and illustrated. The functionality of the ectopic compound eye is discussed. According to label data, larval association with oak rotten log habitats is reported.
Adults of the Neotropical genera Beltia Jacoby (type species: Beltia nicaraguensis Jacoby) and Colaspoides Laporte (type species: Colaspoides limbata [Olivier]) (Chrysomelidae: Eumolpinae: Eumolpini) are difficult to separate. In this paper, the genus Beltia Jacoby is redefined and diagnosed by features of the pygidium, lateral wings of the prosternum, and metatibiae to distinguish it from Colaspoides and other medium-sized, ovate Eumolpini. Fourteen new species from Costa Rica, Panama, Colombia, Ecuador, and Peru are described and illustrated—Beltia awapita, B. confusa, B. gorgona, B. herreri, B. ledesmae, B. napoensis, B. osa, B. rugosa, B. sanchezae, B. talaga, B. tilarana, B. tisingalita, B. tsachila and B. vacilona. A key and range maps for all species recognized herein are provided. Colaspoides placidula Bechyne, Colaspoides placidula angustomarginata Bechyne, Colaspoides chiriquensis Jacoby, and Colaspoides weyrauchi Bechyne are transferred to Beltia and redescribed. Colaspoides turrialbana Bechyne is synonymized with B. chiriquensis, and Colaspoides chanchamaya Bechyne is synonomized with B. weyrauchi. Morphological similarities with Beltia indicate that Old World Colaspoides also should be removed from Colaspoides s. str.
Forrester, Alison ; De Leonibus, Chiara ; Grumati, Paolo ; Fasana, Elisa ; Piemontese, Marilina ; Staiano, Leopoldo ; Fregno, Ilaria ; Raimondi, Andrea ; Marazza, Alessandro ; Bruno, Gemma ; Iavazzo, Maria ; Intartaglia, Daniela ; Seczynska, Marta ; Anken, Eelco van ; Conte, Ivan ; De Matteis, Maria Antonietta ; Đikić, Ivan ; Molinari, Maurizio ; Settembre, Carmine
Autophagy is a cytosolic quality control process that recognizes substrates through receptor‐mediated mechanisms. Procollagens, the most abundant gene products in Metazoa, are synthesized in the endoplasmic reticulum (ER), and a fraction that fails to attain the native structure is cleared by autophagy. However, how autophagy selectively recognizes misfolded procollagens in the ER lumen is still unknown. We performed siRNA interference, CRISPR‐Cas9 or knockout‐mediated gene deletion of candidate autophagy and ER proteins in collagen producing cells. We found that the ER‐resident lectin chaperone Calnexin (CANX) and the ER‐phagy receptor FAM134B are required for autophagy‐mediated quality control of endogenous procollagens. Mechanistically, CANX acts as co‐receptor that recognizes ER luminal misfolded procollagens and interacts with the ER‐phagy receptor FAM134B. In turn, FAM134B binds the autophagosome membrane‐associated protein LC3 and delivers a portion of ER containing both CANX and procollagen to the lysosome for degradation. Thus, a crosstalk between the ER quality control machinery and the autophagy pathway selectively disposes of proteasome‐resistant misfolded clients from the ER.
Hutten, Nadia R. P. W. ; Kuypers, Kim P. C. ; Wel, Janelle van ; Theunissen, Eef L. ; Tönnes, Stefan W. ; Verkes, Robbert Jan ; Ramaekers, Johannes G.
Background: Prospective memory is the ability to recall intended actions or events at the right time or in the right context. While cannabis is known to impair prospective memory, the acute effect of cocaine is unknown. In addition, it is not clear whether changes in prospective memory represent specific alterations in memory processing or result from more general effects on cognition that spread across multiple domains such as arousal and attention.
Aims: The main objective of the study was, therefore, to determine whether drug-induced changes in prospective memory are memory specific or associated with more general drug-induced changes in attention and arousal.
Methods: A placebo-controlled, three-way, cross-over study including 15 regular poly-drug users was set up to test the influence of oral cocaine (300 mg) and vaporised cannabis (300+150 ‘booster’ µg/kg bodyweight) on an event-based prospective memory task. Attentional performance was assessed using a divided attention task and subjective arousal was assessed with the Profile of Mood States questionnaire.
Results: Results showed that cocaine enhanced prospective memory, attention and arousal. Mean performance of prospective memory and attention, as well as levels of arousal were lowest during treatment with cannabis as compared with placebo and cocaine as evinced by a significantly increased trend across treatment conditions. Prospective memory performance was only weakly positively associated to measures of attention and arousal.
Conclusion: Together, these results indicate that cocaine enhancement of prospective memory performance cannot be fully explained by parallel changes in arousal and attention levels, and is likely to represent a direct change in the neural network underlying prospective memory.
Sharma, Sunny ; David Hartmann, Johannes ; Watzinger, Peter ; Klepper, Arvid ; Peifer, Christian ; Kötter, Peter ; Lafontaine, Denis L. J. ; Entian, Karl-Dieter
The entire chemical modification repertoire of yeast ribosomal RNAs and the enzymes responsible for it have recently been identified. Nonetheless, in most cases the precise roles played by these chemical modifications in ribosome structure, function and regulation remain totally unclear. Previously, we demonstrated that yeast Rrp8 methylates m1A645 of 25S rRNA in yeast. Here, using mung bean nuclease protection assays in combination with quantitative RP-HPLC and primer extension, we report that 25S/28S rRNA of S. pombe, C. albicans and humans also contain a single m1A methylation in the helix 25.1. We characterized nucleomethylin (NML) as a human homolog of yeast Rrp8 and demonstrate that NML catalyzes the m1A1322 methylation of 28S rRNA in humans. Our in vivo structural probing of 25S rRNA, using both DMS and SHAPE, revealed that the loss of the Rrp8-catalyzed m1A modification alters the conformation of domain I of yeast 25S rRNA causing translation initiation defects detectable as halfmers formation, likely because of incompetent loading of 60S on the 43S-preinitiation complex. Quantitative proteomic analysis of the yeast Δrrp8 mutant strain using 2D-DIGE, revealed that loss of m1A645 impacts production of specific set of proteins involved in carbohydrate metabolism, translation and ribosome synthesis. In mouse, NML has been characterized as a metabolic disease-associated gene linked to obesity. Our findings in yeast also point to a role of Rrp8 in primary metabolism. In conclusion, the m1A modification is crucial for maintaining an optimal 60S conformation, which in turn is important for regulating the production of key metabolic enzymes.
Karimova, Madina ; Baker, Oliver ; Camgoz, Aylin ; Naumann, Ronald ; Buchholz, Frank ; Anastassiadis, Konstantinos
Site-specific recombinases (SSR) are utilized as important genome engineering tools to precisely modify the genome of mice and other model organisms. Reporter mice that mark cells that at any given time had expressed the enzyme are frequently used for lineage tracing and to characterize newly generated mice expressing a recombinase from a chosen promoter. With increasing sophistication of genome alteration strategies, the demand for novel SSR systems that efficiently and specifically recombine their targets is rising and several SSR-systems are now used in combination to address complex biological questions in vivo. Generation of reporter mice for each one of these recombinases is cumbersome and increases the number of mouse lines that need to be maintained in animal facilities. Here we present a multi-reporter mouse line for loci-of-recombination (X) (MuX) that streamlines the characterization of mice expressing prominent recombinases. MuX mice constitutively express nuclear green fluorescent protein after recombination by either Cre, Flp, Dre or Vika recombinase, rationalizing the number of animal lines that need to be maintained. We also pioneer the use of the Vika/vox system in mice, illustrating its high efficacy and specificity, thereby facilitating future designs of sophisticated recombinase-based in vivo genome engineering strategies.
Many cellular processes are regulated via pH, and maintaining the pH of different organelles is crucial for cell survival. A pH-sensitive GFP variant, the so-called pHluorin, has proven to be a valuable tool to study the pH of the cytosol, mitochondria and other organelles in vivo. We found that the fluorescence intensity of Endoplasmic Reticulum (ER)-targeted pHluorin in the yeast Saccharomyces cerevisiae was very low and barely showed pH sensitivity, probably due to misfolding in the oxidative environment of the ER. We therefore developed a superfolder variant of pHluorin which enabled us to monitor pH changes in the ER and the cytosol of S. cerevisiae in vivo. The superfolder pHluorin variant is likely to be functional in cells of different organisms as well as in additional compartments that originate from the secretory pathway like the Golgi apparatus and pre-vacuolar compartments, and therefore has a broad range of possible future applications.
The males of Caraphia squamosa (Chemsak and Linsley, 1984) and C. seriata (Chemsak and Linsley, 1984), and the female of C. lingafelteri Ohbayashi and Yamasako, 2016 (Coleoptera: Cerambycidae: Lepturinae) are described for the first time. Two new Caraphia species are described: C. warneri Wappes and Santos-Silva, from Guatemala; and C. woodruffi Wappes and Santos-Silva, from Guatemala and Honduras. A key to American species of Caraphia and a map showing their known distribution is provided. New country records for C. seriata and C. lingafelteri are also provided. Lastly, the C. seriata record for Honduras was based on specimens of a new species (Caraphia lingafelteri), hence the Honduras record should be deleted.
Introduction: To date, several meta-analyses clearly demonstrated that resistance and plyometric training are effective to improve physical fitness in children and adolescents. However, a methodological limitation of meta-analyses is that they synthesize results from different studies and hence ignore important differences across studies (i.e., mixing apples and oranges). Therefore, we aimed at examining comparative intervention studies that assessed the effects of age, sex, maturation, and resistance or plyometric training descriptors (e.g., training intensity, volume etc.) on measures of physical fitness while holding other variables constant.
Methods: To identify relevant studies, we systematically searched multiple electronic databases (e.g., PubMed) from inception to March 2018. We included resistance and plyometric training studies in healthy young athletes and non-athletes aged 6 to 18 years that investigated the effects of moderator variables (e.g., age, maturity, sex, etc.) on components of physical fitness (i.e., muscle strength and power).
Results: Our systematic literature search revealed a total of 75 eligible resistance and plyometric training studies, including 5,138 participants. Mean duration of resistance and plyometric training programs amounted to 8.9 ± 3.6 weeks and 7.1±1.4 weeks, respectively. Our findings showed that maturation affects plyometric and resistance training outcomes differently, with the former eliciting greater adaptations pre-peak height velocity (PHV) and the latter around- and post-PHV. Sex has no major impact on resistance training related outcomes (e.g., maximal strength, 10 repetition maximum). In terms of plyometric training, around-PHV boys appear to respond with larger performance improvements (e.g., jump height, jump distance) compared with girls. Different types of resistance training (e.g., body weight, free weights) are effective in improving measures of muscle strength (e.g., maximum voluntary contraction) in untrained children and adolescents. Effects of plyometric training in untrained youth primarily follow the principle of training specificity. Despite the fact that only 6 out of 75 comparative studies investigated resistance or plyometric training in trained individuals, positive effects were reported in all 6 studies (e.g., maximum strength and vertical jump height, respectively).
Conclusions: The present review article identified research gaps (e.g., training descriptors, modern alternative training modalities) that should be addressed in future comparative studies.
Spahn, Christoph Klaus ; Glaesmann, Mathilda ; Grimm, Jonathan B. ; Ayala, Anthony X. ; Lavis, Luke D. ; Heilemann, Mike
Maintenance of the bacterial homeostasis initially emanates from interactions between proteins and the bacterial nucleoid. Investigating their spatial correlation requires high spatial resolution, especially in tiny, highly confined and crowded bacterial cells. Here, we present super-resolution microscopy using a palette of fluorescent labels that bind transiently to either the membrane or the nucleoid of fixed E. coli cells. The presented labels are easily applicable, versatile and allow long-term single-molecule super-resolution imaging independent of photobleaching. The different spectral properties allow for multiplexed imaging in combination with other localisation-based super-resolution imaging techniques. As examples for applications, we demonstrate correlated super-resolution imaging of the bacterial nucleoid with the position of genetic loci, of nascent DNA in correlation to the entire nucleoid, and of the nucleoid of metabolically arrested cells. We furthermore show that DNA- and membrane-targeting labels can be combined with photoactivatable fluorescent proteins and visualise the nano-scale distribution of RNA polymerase relative to the nucleoid in drug-treated E. coli cells.
Diploid transgenic organisms are either hemi- or homozygous. Genetic assays are, therefore, required to identify the genotype. Our AGameOfClones vector concept uses two clearly distinguishable transformation markers embedded in interweaved, but incompatible Lox site pairs. Cre-mediated recombination leads to hemizygous individuals that carry only one marker. In the following generation, heterozygous descendants are identified by the presence of both markers and produce homozygous progeny that are selected by the lack of one marker. We prove our concept in Tribolium castaneum by systematically creating multiple functional homozygous transgenic lines suitable for long-term fluorescence live imaging. Our approach saves resources and simplifies transgenic organism handling. Since the concept relies on the universal Cre-Lox system, it is expected to work in all diploid model organisms, for example, insects, zebrafish, rodents and plants. With appropriate adaptions, it can be used in knock-out assays to preselect homozygous individuals and thus minimize the number of wasted animals.
When mapping eye-movement behavior to the visual information presented to an observer, Areas of Interest (AOIs) are commonly employed. For static stimuli (screen without moving elements), this requires that one AOI set is constructed for each stimulus, a possibility in most eye-tracker manufacturers' software. For moving stimuli (screens with moving elements), however, it is often a time-consuming process, as AOIs have to be constructed for each video frame. A popular use-case for such moving AOIs is to study gaze behavior to moving faces. Although it is technically possible to construct AOIs automatically, the standard in this field is still manual AOI construction. This is likely due to the fact that automatic AOI-construction methods are (1) technically complex, or (2) not effective enough for empirical research. To aid researchers in this field, we present and validate a method that automatically achieves AOI construction for videos containing a face. The fully-automatic method uses an open-source toolbox for facial landmark detection, and a Voronoi-based AOI-construction method. We compared the position of AOIs obtained using our new method, and the eye-tracking measures derived from it, to a recently published semi-automatic method. The differences between the two methods were negligible. The presented method is therefore both effective (as effective as previous methods), and efficient; no researcher time is needed for AOI construction. The software is freely available from https://osf.io/zgmch/.
Alten, Rieke ; Burkhardt, Harald ; Feist, Eugen ; Krüger, Klaus ; Rech, Hans Jürgen ; Rubbert-Roth, Andrea ; Voll, Reinhard ; Elbez, Yedid ; Rauch, Christiane
Background: Methotrexate (MTX) remains the anchor drug in rheumatoid arthritis (RA) treatment, but is poorly tolerated or contraindicated in some patients. There is a wealth of data supporting the use of abatacept in combination with MTX, but data on alternative conventional synthetic disease-modifying antirheumatic drug (csDMARD) combinations with abatacept are scarce.
Methods: In this post-hoc exploratory analysis, efficacy and safety data were extracted from abatacept RA studies in which combination with csDMARDs other than MTX was permitted: three interventional trials (ATTAIN, ASSURE, and ARRIVE) and one real-world study (ACTION). Patients with moderate-to-severe RA received abatacept in combination with MTX, hydroxychloroquine, sulfasalazine, azathioprine, or leflunomide for 6 months to 2 years according to the study design. Change from baseline in physical function (Health Assessment Questionnaire—Disability Index (HAQ-DI); all studies) and 28-joint Disease Activity Score (C-reactive protein) (DAS28 (CRP); ATTAIN, ARRIVE, and ACTION), American College of Rheumatology response rates (ATTAIN), and safety were assessed for individual and pooled csDMARD combinations for each trial. A meta-analysis was also performed on pooled data for HAQ-DI and DAS28 (CRP) across interventional trials.
Results: Across all four studies, 731 patients received abatacept plus one non-MTX csDMARD (hydroxychloroquine n = 152; sulfasalazine n = 123; azathioprine n = 59; and leflunomide n = 397) and 2382 patients received abatacept plus MTX. Mean changes from baseline in HAQ-DI scores for abatacept plus MTX (all csDMARDs pooled) vs abatacept plus a non-MTX csDMARD were –0.54 vs –0.44 (ATTAIN), –0.43 vs –0.43 (ASSURE), and –0.39 vs –0.36 (ARRIVE). Mean changes from baseline in DAS28 (CRP) and ACR response rates were also similar with abatacept plus MTX or non-MTX csDMARDs. Data for individual non-MTX csDMARDs (pooled across studies) and real-world data were consistent with these findings. Rates of treatment-related adverse events and serious adverse events, respectively, for abatacept plus one non-MTX csDMARD vs abatacept plus MTX were 35.7% vs 41.7% and 2.4% vs 2.3% (ATTAIN), 58.0% vs 55.9% and 4.2% vs 1.7% (ASSURE), and 38.1% vs 44.3% and 0.6% vs 2.9% (ARRIVE).
Conclusions: Abatacept in combination with non-MTX csDMARDs is clinically effective and well tolerated in patients with moderate-to-severe RA, providing similar benefits to those seen with abatacept plus MTX.
Background: Stem cell therapy is considered as a promising alternative to treat intervertebral disc degeneration (IDD). Extensive work had been done on identifying and comparing different types of candidate stem cells, both in vivo and in vitro. However, few studies have shed light on degenerative nucleus pulposus cells (NPCs), especially their biological behavior under the influence of exogenous stem cells, specifically the gene expression and regulation pattern. In the present study, we aimed to determine messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs), which are differentially expressed during the co-culturing process with adipose-derived mesenchymal stem cells (ASCs) and to explore the involved signaling pathways and the regulatory networks.
Methods: We compared degenerative NPCs co-cultured with ASCs with those cultured solely using lncRNA-mRNA microarray analysis. Based on these data, we investigated the significantly regulated signaling pathways based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database. Moreover, 23 micro RNAs (miRNAs), which were demonstrated to be involved in IDD were chosen; we investigated their theoretic regulatory importance associated with our microarray data.
Results: We found 632 lncRNAs and 1682 mRNAs were differentially expressed out of a total of 40,716 probes. We then confirmed the microarray data by real-time PCR. Furthermore, we demonstrated 197 upregulated, and 373 downregulated Gene Ontology terms and 176 significantly enriched pathways, such as the mitogen-activated protein kinase (MAPK) pathway. Also, a signal-net was constructed to reveal the interplay among differentially expressed genes. Meanwhile, a mRNA-lncRNA co-expression network was constructed for the significantly changed mRNAs and lncRNAs. Also, the competing endogenous RNA (ceRNA) network was built.
Conclusion: Our results present the first comprehensive identification of differentially expressed lncRNAs and mRNAs of degenerative NPCs, altered by co-culturing with ASCs, and outline the gene expression regulation pattern. These may provide valuable information for better understanding of stem cell therapy and potential candidate biomarkers for IDD treatment.
Mücke, Marcus Maximilian ; Mücke, Victoria Therese ; Peiffer, Kai-Henrik ; Sarrazin, Christoph ; Zeuzem, Stefan ; Berger, Annemarie ; Vermehren, Johannes
Background: Patients with chronic hepatitis C virus (HCV) infection and active or previous hepatitis B virus (HBV) are at risk of HBV reactivation (HBV-R) during direct-acting antiviral (DAA) therapy. Recent reports suggest that HBV-R may even occur several months after completion of DAA therapy. The aim of this study was to assess the risk of HBV-R in patients with resolved HBV after successful DAA therapy during long-term follow-up (FU).
Methods: Among 848 patients treated for chronic HCV, all patients with resolved HBV and long-term FU data were eligible for inclusion. Patients were HBV DNA/hepatitis B surface antigen (HBsAg)–negative at the end of therapy (EOT) and were followed for up to 52 weeks thereafter. Patients underwent regular alanine transaminase (ALT) testing, and additional HBV DNA/HBsAg testing was performed at FU week 12, end of FU, and in case of an ALT increase above the upper limit of normal (>ULN).
Results: A total of 108 patients were followed up for a mean (range) of 41.5 (24–52) weeks after EOT. None of the patients experienced reverse HBsAg seroconversion or reappearance of HBV DNA. One patient received a liver transplantation; 1 patient was diagnosed with de novo hepatocellular carcinoma, and 2 patients died. Eighteen patients (16.7%) had increased ALT levels (grade 0/1). Of those, the majority were male (72.2%) and significantly more patients had cirrhosis (66.7% vs 36.2%, P = .015) or received ribavirin as part of their treatment regimen (86.7% vs 46.8%, P = .041). None of these were associated with HBV-R.
Conclusions: Our results indicate that the risk of HBV-R in patients with resolved HBV treated with DAAs for HCV is low during long-term follow-up.
Al-Shahi Salman, Rustam ; Frantzias, Joseph ; Lee, Robert J. ; Lyden, Patrick D. ; Battey, Thomas W. K. ; Ayres, Alison M. ; Goldstein, Joshua Norkin ; Mayer, Stephan A. ; Steiner, Thorsten ; Wang, Xia ; Arima, Hisatomi ; Hasegawa, Hitoshi ; Oishi, Makoto ; Godoy, Daniel Agustín ; Masotti, Luca ; Dowlatshahi, Dar ; Rodríguez-Luna, David ; Molina, Carlos ; Jang, Dong-Kyu ; Davalos, Antonio ; Castillo, José ; Yao, Xiaoying ; Claassen, Jan ; Volbers, Bastian ; Kazui, Seiji ; Okada, Yasushi ; Fujimoto, Shigeru ; Toyoda, Kazunori ; Li, Qi ; Khoury, Jane ; Delgado, Pilar ; Álvarez-Sabín, José ; Hernández-Guillamon, Mar ; Prats-Sánchez, Luis ; Cai, Chunyan ; Kate, Mahesh ; McCourt, Rebecca ; Venkatasubramanian, Chitra ; Diringer, Michael N. ; Ikeda, Yukio ; Worthmann, Hans ; Ziai, Wendy C. ; D’Esterre, Christopher D. ; Aviv, Richard I. ; Raab, Peter ; Murai, Yasuo ; Zazulia, Allyson R. ; Butcher, Kenneth ; Seyedsaadat, Seyed Mohammad ; Grotta, James C. ; Martí-Fàbregas, Joan ; Montaner, Joan ; Broderick, Joseph ; Yamamoto, Haruko ; Staykov, Dimitre ; Connolly, Edward Sander ; Selim, Magdy ; Leira, Rogelio ; Moon, Byung Hoo ; Demchuk, Andrew M. ; Di Napoli, Mario ; Fujii, Yukihiko ; Anderson, Craig S. ; Rosand, Jonathan
Background: Intracerebral haemorrhage growth is associated with poor clinical outcome and is a therapeutic target for improving outcome. We aimed to determine the absolute risk and predictors of intracerebral haemorrhage growth, develop and validate prediction models, and evaluate the added value of CT angiography.
Methods: In a systematic review of OVID MEDLINE—with additional hand-searching of relevant studies' bibliographies— from Jan 1, 1970, to Dec 31, 2015, we identified observational cohorts and randomised trials with repeat scanning protocols that included at least ten patients with acute intracerebral haemorrhage. We sought individual patient-level data from corresponding authors for patients aged 18 years or older with data available from brain imaging initially done 0·5–24 h and repeated fewer than 6 days after symptom onset, who had baseline intracerebral haemorrhage volume of less than 150 mL, and did not undergo acute treatment that might reduce intracerebral haemorrhage volume. We estimated the absolute risk and predictors of the primary outcome of intracerebral haemorrhage growth (defined as >6 mL increase in intracerebral haemorrhage volume on repeat imaging) using multivariable logistic regression models in development and validation cohorts in four subgroups of patients, using a hierarchical approach: patients not taking anticoagulant therapy at intracerebral haemorrhage onset (who constituted the largest subgroup), patients taking anticoagulant therapy at intracerebral haemorrhage onset, patients from cohorts that included at least some patients taking anticoagulant therapy at intracerebral haemorrhage onset, and patients for whom both information about anticoagulant therapy at intracerebral haemorrhage onset and spot sign on acute CT angiography were known.
Findings: Of 4191 studies identified, 77 were eligible for inclusion. Overall, 36 (47%) cohorts provided data on 5435 eligible patients. 5076 of these patients were not taking anticoagulant therapy at symptom onset (median age 67 years, IQR 56–76), of whom 1009 (20%) had intracerebral haemorrhage growth. Multivariable models of patients with data on antiplatelet therapy use, data on anticoagulant therapy use, and assessment of CT angiography spot sign at symptom onset showed that time from symptom onset to baseline imaging (odds ratio 0·50, 95% CI 0·36–0·70; p<0·0001), intracerebral haemorrhage volume on baseline imaging (7·18, 4·46–11·60; p<0·0001), antiplatelet use (1·68, 1·06–2·66; p=0·026), and anticoagulant use (3·48, 1·96–6·16; p<0·0001) were independent predictors of intracerebral haemorrhage growth (C-index 0·78, 95% CI 0·75–0·82). Addition of CT angiography spot sign (odds ratio 4·46, 95% CI 2·95–6·75; p<0·0001) to the model increased the C-index by 0·05 (95% CI 0·03–0·07).
Interpretation: In this large patient-level meta-analysis, models using four or five predictors had acceptable to good discrimination. These models could inform the location and frequency of observations on patients in clinical practice, explain treatment effects in prior randomised trials, and guide the design of future trials.
Funding: UK Medical Research Council and British Heart Foundation.
We evaluate the near-surface representation of thermally driven winds in the Swiss Alps in a numerical weather prediction model at km-scale resolution. In addition, the influence of grid resolution (2.2 km and 1.1 km), topography filtering, and land surface datasets on the accuracy of the simulated valley winds is investigated. The simulations are evaluated against a comprehensive set of surface observations for an 18-day fair-weather summer period in July 2006. The episode is characterized by strong diurnal wind systems and the formation of shallow convection over the mountains, which transitions to precipitating convection in some areas. The near-surface winds (10 m above ground level) follow a typical diurnal pattern with strong daytime up-valley flow and weaker nighttime down-valley flow. At a 2.2 km resolution the valley winds are poorly simulated for most stations, while at a 1.1 km resolution the diurnal cycle of the valley winds is well represented in most large (e.g., Rhein valley at Chur and Rhone valley at Visp) and medium-sized valleys (e.g., Linth valley at Glarus). In the smaller valleys (e.g., Maggia valley at Cevio), the amplitude of the valley wind is still significantly underestimated, even at a 1.1 km resolution. Detailed sensitivity experiments show that the use of high-resolution land surface datasets, for both the soil characteristics as well as for the land cover, and reduced filtering of the topography are essential to achieve good performance at a 1.1 km resolution
Glatzel, Daniel Karl ; Koeberle, Andreas ; Pein, Helmut ; Löser, Konstantin ; Stark, Anna ; Keksel, Nelli ; Werz, Oliver ; Müller, Rolf ; Bischoff, Iris ; Fürst, Robert
The enzyme acetyl-CoA carboxylase (ACC) plays a crucial role in fatty acid metabolism. In recent years, ACC has been recognized as a promising drug target for treating different diseases. However, the role of ACC in vascular endothelial cells (ECs) has been neglected so far. To characterize the role of ACC, we used the ACC inhibitor, soraphen A, as a chemical tool, and also a gene silencing approach. We found that ACC1 was the predominant isoform in human umbilical vein ECs as well as in human microvascular ECs and that soraphen A reduced the levels of malonyl-CoA. We revealed that ACC inhibition shifted the lipid composition of EC membranes. Accordingly, membrane fluidity, filopodia formation, and migratory capacity were reduced. The antimigratory action of soraphen A depended on an increase in the cellular proportion of PUFAs and, most importantly, on a decreased level of phosphatidylglycerol. Our study provides a causal link between ACC, membrane lipid composition, and cell migration in ECs. Soraphen A represents a useful chemical tool to investigate the role of fatty acid metabolism in ECs and ACC inhibition offers a new and valuable therapeutic perspective for the treatment of EC migration-related diseases.