bluuebunny/crossref_metadata_2025 huggingface dataset:
Structural basis for substrate specificity and catalysis of α1,6-fucosyltransferase
Authors: García-García Ana , Ceballos-Laita Laura , Serna Sonia , Artschwager Raik , Reichardt Niels C. , Corzana Francisco , Hurtado-Guerrero Ramon
Published: February 2020

Core-fucosylation is an essential biological modification by which a fucose is transferred from GDP-β-L-fucose to the innermost N-acetylglucosamine residue of N-linked glycans. A single human enzyme α1,6-fucosyltransferase (FUT8) is the only enzyme responsible for this modification via the addition of an α-1,6-linked fucose to N-glycans. To date, the details of substrate recognition and catalysis by FUT8 remain unknown. Here, we report the crystal structure of FUT8 complexed with GDP and a biantennary complex N-glycan (G0), which provides insight into both substrate recognition and catalysis. FUT8 follows an SN2 mechanism and deploys a series of loops and an α-helix which all contribute in forming the binding site. An exosite, formed by one of these loops and an SH3 domain, is responsible for the recognition of branched sugars, making contacts specifically to the α1,3 arm GlcNAc, a feature required for catalysis. This information serves as a framework for inhibitor design, and helps to assess its potential as a therapeutic target.
Gaze, behavioral, and clinical data for phantom limbs after hand amputation from 15 amputees and 29 controls
Authors: Saetta Gianluca , Cognolato Matteo , Atzori Manfredo , Faccio Diego , Giacomino Katia , Mittaz Hager Anne-Gabrielle , Tiengo Cesare , Bassetto Franco , Müller Henning , Brugger Peter
Published: February 2020

Despite recent advances in prosthetics, many upper limb amputees still use prostheses with some reluctance. They often do not feel able to incorporate the artificial hand into their bodily self. Furthermore, prosthesis fitting is not usually tailored to accommodate the characteristics of an individual’s phantom limb sensations. These are experienced by almost all persons with an acquired amputation and comprise the motor and postural properties of the lost limb. This article presents and validates a multimodal dataset including an extensive qualitative and quantitative assessment of phantom limb sensations in 15 transradial amputees, surface electromyography and accelerometry data of the forearm, and measurements of gaze behavior during exercises requiring pointing or repositioning of the forearm and the phantom hand. The data also include acquisitions from 29 able-bodied participants, matched for gender and age. Special emphasis was given to tracking the visuo-motor coupling between eye-hand/eye-phantom during these exercises.
Pollen defenses negatively impact foraging and fitness in a generalist bee (Bombus impatiens: Apidae)
Authors: Brochu Kristen K. , van Dyke Maria T. , Milano Nelson J. , Petersen Jessica D. , McArt Scott H. , Nault Brian A. , Kessler André , Danforth Bryan N.
Published: February 2020

Plants may benefit from limiting the community of generalist floral visitors if the species that remain are more effective pollinators and less effective pollenivores. Plants can reduce access to pollen through altered floral cues or morphological structures, but can also reduce consumption through direct pollen defenses. We observed thatEucera (Peponapis) pruinosa, a specialist bee onCucurbitaplants, collected pure loads of pollen while generalist honey bees and bumble bees collected negligible amounts of cucurbit pollen, even though all groups of bees visited these flowers. Cucurbit flowers have no morphological adaptations to limit pollen collection by bees, thus we assessed their potential for physical, nutritional, and chemical pollen traits that might act as defenses to limit pollen loss to generalist pollinators. Bumble bee (Bombus impatiens) microcolonies experienced reduced pollen consumption, mortality, and reproduction as well as increased stress responses when exposed to nutritional and mechanical pollen defenses. These bees also experienced physiological effects of these defenses in the form of hindgut expansion and gut melanization. Chemical defenses alone increased the area of gut melanization in larger bees and induced possible compensatory feeding. Together, these results suggest that generalist bumble bees avoid collecting cucurbit pollen due to the physiological costs of physical and chemical pollen defenses.
Phase I studies of vorinostat with ixazomib or pazopanib imply a role of antiangiogenesis-based therapy for TP53 mutant malignancies
Authors: Wang Yudong , Janku Filip , Piha-Paul Sarina , Hess Kenneth , Broaddus Russell , Liu Lidong , Shi Naiyi , Overman Michael , Kopetz Scott , Subbiah Vivek , Naing Aung , Hong David , Tsimberidou Apostolia M. , Karp Daniel , Yao James , Fu Siqing
Published: February 2020

We performed two phase I trials of the histone deacetylase inhibitor vorinostat combined with either the vascular endothelial growth factor inhibitor pazopanib (NCT01339871) or the proteasome inhibitor ixazomib (NCT02042989) in patients with metastatic TP53 mutant solid tumors. Both trials followed a 3 + 3 dose-escalation design allowing for a dose expansion cohort of up to 14 additional patients with a specific tumor type. Patients had to have a confirmed TP53 mutation to be enrolled in NCT02042989. Among patients enrolled in NCT01339871, TP53 mutation status was determined for those for whom tumor specimens were available. The results of NCT01339871 were reported previously. Common treatment-related adverse events in NCT02042989 included anemia, thrombocytopenia, fatigue, nausea, vomiting, and diarrhea. Compared with patients with metastatic TP53 hotspot mutant solid tumors who were treated with ixazomib and vorinostat (n = 59), those who were treated with pazopanib and vorinostat (n = 11) had a significantly higher rate of clinical benefit, defined as stable disease lasting ≥6 months or an objective response (3.4% vs. 45%; p < 0.001), a significantly longer median progression-free survival duration (1.7 months [95% confidence interval (CI), 1.1–2.3] vs. 3.5 months [95% CI, 1.7–5.2]; p = 0.002), and a longer median overall survival duration (7.3 months [95% CI, 4.8–9.8] vs. 12.7 months [95% CI, 7.1–18.3]; p = 0.24). Our two phase I trials provide preliminary evidence supporting the use of antiangiogenisis-based therapy in patients with metastatic TP53 mutant solid tumors, especially in those with metastatic sarcoma or metastatic colorectal cancer.
A tiny new Middle Triassic stem-lepidosauromorph from Germany: implications for the early evolution of lepidosauromorphs and the Vellberg fauna
Authors: Sobral Gabriela , Simões Tiago R. , Schoch Rainer R.
Published: February 2020

The Middle Triassic was a time of major changes in tetrapod faunas worldwide, but the fossil record for this interval is largely obscure for terrestrial faunas. This poses a severe limitation to our understanding on the earliest stages of diversification of lineages representing some of the most diverse faunas in the world today, such as lepidosauromorphs (e.g., lizards and tuataras). Here, we report a tiny new lepidosauromorph from the Middle Triassic from Vellberg (Germany), which combines a mosaic of features from both early evolving squamates and rhynchocephalians, such as the simultaneous occurrence of a splenial bone and partial development of acrodonty. Phylogenetic analyses applying different optimality criteria, and combined morphological and molecular data, consistently recover the new taxon as a stem-lepidosauromorph, implying stem-lepidosauromorph species coinhabited areas comprising today’s central Europe at the same time as the earliest known rhynchocephalians and squamates. It further demonstrates a more complex evolutionary scenario for dental evolution in early lepidosauromorphs, with independent acquisitions of acrodonty early in their evolutionary history. The small size of most terrestrial vertebrates from Vellberg is conspicuous, contrasting to younger Triassic deposits worldwide, but comparable to Early Triassic faunas, suggesting a potential long-lasting Lilliput effect in this fauna.
Neural correlates of subjective timing precision and confidence
Authors: Arnold Derek H. , Hohaia Wiremu , Yarrow Kielan
Published: February 2020

Humans perceptual judgments are imprecise, as repeated exposures to the same physical stimulation (e.g. audio-visual inputs separated by a constant temporal offset) can result in different decisions. Moreover, there can be marked individual differences – precise judges will repeatedly make the same decision about a given input, whereas imprecise judges will make different decisions. The causes are unclear. We examined this using audio-visual (AV) timing and confidence judgments, in conjunction with electroencephalography (EEG) and multivariate pattern classification analyses. One plausible cause of differences in timing precision is that it scales with variance in the dynamics of evoked brain activity. Another possibility is that equally reliable patterns of brain activity are evoked, but there are systematic differences that scale with precision. Trial-by-trial decoding of input timings from brain activity suggested precision differences may not result from variable dynamics. Instead, precision was associated with evoked responses that were exaggerated (more different from baseline) ~300 ms after initial physical stimulations. We suggest excitatory and inhibitory interactions within a winner-take-all neural code for AV timing might exaggerate responses, such that evoked response magnitudes post-stimulation scale with encoding success.
The juvenile hormone described in Rhodnius prolixus by Wigglesworth is juvenile hormone III skipped bisepoxide
Authors: Villalobos-Sambucaro Maria Jose , Nouzova Marcela , Ramirez Cesar E. , Eugenia Alzugaray María , Fernandez-Lima Francisco , Ronderos Jorge Rafael , Noriega Fernando G.
Published: February 2020

Juvenile hormones (JHs) are sesquiterpenoids synthesized by the corpora allata (CA). They play critical roles during insect development and reproduction. The first JH was described in 1934 as a “metamorphosis inhibitory hormone” in Rhodnius prolixus by Sir Vincent B. Wigglesworth. Remarkably, in spite of the importance of R. prolixus as vectors of Chagas disease and model organisms in insect physiology, the original JH that Wigglesworth described for the kissing-bug R. prolixus remained unidentified. We employed liquid chromatography mass spectrometry to search for the JH homologs present in the hemolymph of fourth instar nymphs of R. prolixus. Wigglesworth’s original JH is the JH III skipped bisepoxide (JHSB3), a homolog identified in other heteropteran species. Changes in the titer of JHSB3 were studied during the 10-day long molting cycle of 4th instar nymph, between a blood meal and the ecdysis to 5th instar. In addition we measured the changes of mRNA levels in the CA for the 13 enzymes of the JH biosynthetic pathway during the molting cycle of 4th instar. Almost 90 years after the first descriptions of the role of JH in insects, this study finally reveals that the specific JH homolog responsible for Wigglesworth’s original observations is JHSB3.
ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab
Authors: Agnolon Valentina , Contato Anna , Meneghello Anna , Tagliabue Elda , Toffoli Giuseppe , Gion Massimo , Polo Federico , Fabricio Aline S. C.
Published: February 2020

Circulating HER2 extracellular domain (HER2 ECD) levels were proposed as a surrogate for HER2 tissue expression to monitor breast cancer patients for early relapse or responses to standard or HER2-targeted therapies, such as the monoclonal antibody (mAb) trastuzumab. Currently, available commercial ELISA assays for HER2 ECD rely on antibodies recognizing undisclosed or unknown epitopes. In this work, two ELISA assays employing MGR2 and MGR3 epitope-specific mAbs for HER2 ECD were developed and validated, showing good assay precision and linearity of the dose-response signal within the dynamic range of 0.19–12.50 ng mL−1 and detection limits of 0.76 and 0.75 ng mL−1 for the MGR2 and MGR3 assays, respectively. The developed assay showed a good agreement with two widely used commercial kits for HER2 ECD quantification in serum samples from breast cancer patients. A complete characterization of mAb-HER2 ECD interaction was performed by means of surface plasmon resonance using trastuzumab as control for both epitope mapping and kinetics analysis. The epitopes recognized by the two mAbs showed no overlap with trastuzumab, which was confirmed by trastuzumab interference analysis in serum samples. The method showed to be a practical approach to determine HER2 ECD with a high degree of sensitivity, reliability and recovery in samples containing mAbs-based therapies.
Inhibition of IL-1beta improves Glycaemia in a Mouse Model for Gestational Diabetes
Authors: Schulze Friederike , Wehner Josua , Kratschmar Denise V. , Makshana Valmir , Meier Daniel T. , Häuselmann Stéphanie P. , Dalmas Elise , Thienel Constanze , Dror Erez , Wiedemann Sophia J. , Traub Shuyang , Nordmann Thierry M. , Rachid Leila , De Baat Axel , Rohm Theresa V. , Zhao Cheng , Odermatt Alex , Böni-Schnetzler Marianne , Donath Marc Y.
Published: February 2020

Gestational diabetes mellitus (GDM) is one of the most common diseases associated with pregnancy, however, the underlying mechanisms remain unclear. Based on the well documented role of inflammation in type 2 diabetes, the aim was to investigate the role of inflammation in GDM. We established a mouse model for GDM on the basis of its two major risk factors, obesity and aging. In these GDM mice, we observed increased Interleukin-1β (IL-1β) expression in the uterus and the placenta along with elevated circulating IL-1β concentrations compared to normoglycemic pregnant mice. Treatment with an anti-IL-1β antibody improved glucose-tolerance of GDM mice without apparent deleterious effects for the fetus. Finally, IL-1β antagonism showed a tendency for reduced plasma corticosterone concentrations, possibly explaining the metabolic improvement. We conclude that IL-1β is a causal driver of impaired glucose tolerance in GDM.
Denitrification rates in lake sediments of mountains affected by high atmospheric nitrogen deposition
Authors: Palacin-Lizarbe Carlos , Camarero Lluís , Hallin Sara , Jones Christopher M. , Catalan Jordi
Published: February 2020

During the last decades, atmospheric nitrogen loading in mountain ranges of the Northern Hemisphere has increased substantially, resulting in high nitrate concentrations in many lakes. Yet, how increased nitrogen has affected denitrification, a key process for nitrogen removal, is poorly understood. We measured actual and potential (nitrate and carbon amended) denitrification rates in sediments of several lake types and habitats in the Pyrenees during the ice-free season. Actual denitrification rates ranged from 0 to 9 μmol N2O m−2 h−1 (mean, 1.5 ± 1.6 SD), whereas potential rates were about 10-times higher. The highest actual rates occurred in warmer sediments with more nitrate available in the overlying water. Consequently, littoral habitats showed, on average, 3-fold higher rates than the deep zone. The highest denitrification potentials were found in more productive lakes located at relatively low altitude and small catchments, with warmer sediments, high relative abundance of denitrification nitrite reductase genes, and sulphate-rich waters. We conclude that increased nitrogen deposition has resulted in elevated denitrification rates, but not sufficiently to compensate for the atmospheric nitrogen loading in most of the highly oligotrophic lakes. However, there is potential for high rates, especially in the more productive lakes and landscape features largely govern this.
Decimated little brown bats show potential for adaptive change
Authors: Auteri Giorgia G. , Knowles L. Lacey
Published: February 2020

The degree to which species can rapidly adapt is key to survival in the face of climatic and other anthropogenic changes. For little brown bats (Myotis lucifugus), whose populations have experienced declines of over 90% because of the introduced fungal pathogen that causes white-nose syndrome (WNS), survival of the species may ultimately depend upon its capacity for adaptive change. Here, we present evidence of selectively driven change (adaptation), despite dramatic nonadaptive genomic shifts (genetic drift) associated with population declines. We compared the genetic makeups of wild survivors versus non-survivors of WNS, and found significant shifts in allele frequencies of genes associated with regulating arousal from hibernation (GABARB1), breakdown of fats (cGMP-PK1), and vocalizations (FOXP2). Changes at these genes are suggestive of evolutionary adaptation, given that WNS causes bats to arouse with unusual frequency from hibernation, contributing to premature depletion of fat reserves. However, whether these putatively adaptive shifts in allele frequencies translate into sufficient increases in survival for the species to rebound in the face of WNS is unknown.
Meta-analysis of honey bee neurogenomic response links Deformed wing virus type A to precocious behavioral maturation
Authors: Traniello Ian M. , Bukhari Syed Abbas , Kevill Jessica , Ahmed Amy Cash , Hamilton Adam R. , Naeger Nicholas L. , Schroeder Declan C. , Robinson Gene E.
Published: February 2020

Crop pollination by the western honey bee Apis mellifera is vital to agriculture but threatened by alarmingly high levels of colony mortality, especially in Europe and North America. Colony loss is due, in part, to the high viral loads of Deformed wing virus (DWV), transmitted by the ectoparasitic mite Varroa destructor, especially throughout the overwintering period of a honey bee colony. Covert DWV infection is commonplace and has been causally linked to precocious foraging, which itself has been linked to colony loss. Taking advantage of four brain transcriptome studies that unexpectedly revealed evidence of covert DWV-A infection, we set out to explore whether this effect is due to DWV-A mimicking naturally occurring changes in brain gene expression that are associated with behavioral maturation. Consistent with this hypothesis, we found that brain gene expression profiles of DWV-A infected bees resembled those of foragers, even in individuals that were much younger than typical foragers. In addition, brain transcriptional regulatory network analysis revealed a positive association between DWV-A infection and transcription factors previously associated with honey bee foraging behavior. Surprisingly, single-cell RNA-Sequencing implicated glia, not neurons, in this effect; there are relatively few glial cells in the insect brain and they are rarely associated with behavioral plasticity. Covert DWV-A infection also has been linked to impaired learning, which together with precocious foraging can lead to increased occurrence of infected bees from one colony mistakenly entering another colony, especially under crowded modern apiary conditions. These findings provide new insights into the mechanisms by which DWV-A affects honey bee health and colony survival.
Chromosome 19 microRNA cluster enhances cell reprogramming by inhibiting epithelial-to-mesenchymal transition
Authors: Mong Ezinne F. , Yang Ying , Akat Kemal M. , Canfield John , VanWye Jeffrey , Lockhart John , Tsibris John C. M. , Schatz Frederick , Lockwood Charles J. , Tuschl Thomas , Kayisli Umit A. , Totary-Jain Hana
Published: February 2020

During implantation, cytotrophoblasts undergo epithelial-to-mesenchymal transition (EMT) as they differentiate into invasive extravillous trophoblasts (EVTs). The primate-specific microRNA cluster on chromosome 19 (C19MC) is exclusively expressed in the placenta, embryonic stem cells and certain cancers however, its role in EMT gene regulation is unknown. In situ hybridization for miR-517a/c, a C19MC cistron microRNA, in first trimester human placentas displayed strong expression in villous trophoblasts and a gradual decrease from proximal to distal cell columns as cytotrophoblasts differentiate into invasive EVTs. To investigate the role of C19MC in the regulation of EMT genes, we employed the CRISPR/dCas9 Synergistic Activation Mediator (SAM) system, which induced robust transcriptional activation of the entire C19MC cistron and resulted in suppression of EMT associated genes. Exposure of human iPSCs to hypoxia or differentiation of iPSCs into either cytotrophoblast-stem-like cells or EVT-like cells under hypoxia reduced C19MC expression and increased EMT genes. Furthermore, transcriptional activation of the C19MC cistron induced the expression of OCT4 and FGF4 and accelerated cellular reprogramming. This study establishes the CRISPR/dCas9 SAM as a powerful tool that enables activation of the entire C19MC cistron and uncovers its novel role in suppressing EMT genes critical for maintaining the epithelial cytotrophoblasts stem cell phenotype.
Si2Ge: A New VII-Type Clathrate with Ultralow Thermal Conductivity and High Thermoelectric Property
Authors: Shen Jinni , Xie Tianzhu , Zhang Longkun , Wang Ping , Fang Zhenxing
Published: February 2020

Based on global particle-swarm optimization algorithm and density functional theory methods, we predicted an alloyed Si2Ge compond with body centered tetragonal type VII clathrate (space group I4/mmm) built by a truncated octahedron fromed by six quadrangles and eight hexagons ([4668]). Si2Ge clathrate is 0.06 eV/atom lower than VII Si clathrate and thermally stable up to 1000 K. It has an indirect band gap of 0.23 eV, high p-doping Seebeck coefficient and n-doping electrical conductivity. It owns a low lattice thermal conductivity of 0.28 W/mK at 300 K because of its weak bonding and strong anharmonic interaction of longitudinal acoustic and low-lying optical phonons. The moderate electronic transport properties together with low lattice thermal conductivity results in a high optimal thermoeletric performance value of 2.54 (1.49) at 800 (1000) K in n (p)-doped Si2Ge.
Egg genotyping reveals the possibility of patent Ancylostoma caninum infection in human intestine
Authors: Furtado Luis Fernando Viana , Dias Lucas Teixeira de Oliveira , Rodrigues Thais de Oliveira , Silva Vivian Jordania da , Oliveira Valéria Nayara Gomes Mendes de , Rabelo Élida Mara Leite
Published: February 2020

Hookworms are intestinal parasites that cause major public health problems, especially in developing countries. To differentiate eggs from different hookworm species, it is necessary to use molecular methodologies, since the eggs are morphologically similar. Here, we performed the molecular identification of single hookworm eggs from six Brazilian states. Of the 634 eggs individually analyzed, 98.1% (622/634) represented Necator americanus, and surprisingly, 1.9% (12/634 eggs from the same patient) represented Ancylostoma caninum. DNA analysis of the A. caninum-positive stool sample revealed no contamination with animal feces. This is the first report of the presence of A. caninum eggs in human feces, which may have a direct implication for the epidemiology of hookworm infection caused by this species. This suggests the need for special attention regarding prophylaxis, as different reservoirs, previously not described, may have great relevance for the spread of A. caninum.
Delegated Regressor, A Robust Approach for Automated Anomaly Detection in the Soil Radon Time Series Data
Authors: Rafique Muhammad , Tareen Aleem Dad Khan , Mir Adil Aslim , Nadeem Malik Sajjad Ahmed , Asim Khawaja M. , Kearfott Kimberlee Jane
Published: February 2020

We propose a new method based on the idea of delegating regressors for predicting the soil radon gas concentration (SRGC) and anomalies in radon or any other time series data. The proposed method is compared to different traditional boosting e.g., Extreme Gradient Boosting (EGB) and simple regression methods e.g., support vector regressors with linear kernel and radial kernel in terms of accurate predictions. R language has been used for the statistical analysis of radon time series (RTS) data. The results obtained show that the proposed methodology predicts SRGC more accurately when compared to different traditional boosting and regression methods. The best correlation is found between the actual and predicted radon concentration for window size of 2 i.e., two days before and after the start of seismic activities. RTS data was collected from 05 February 2017 to 16 February 2018, including 7 seismic events recorded during the study period. Findings of study show that the proposed methodology predicts the SRGC with more precision, for all the window sizes, by overlapping predicted with the actual radon time series concentrations.
The Proteolytic Fraction From Vasconcellea cundinamarcensis Latex Displays Anti-Inflammatory Effect in A Mouse Model of Acute TNBS-Induced Colitis
Authors: Albuquerque Ronniel Morais , Pizzitola Marina Passos , Araújo e Silva Ana Cândida , Dittz Dalton , Freitas Kátia Michelle , Ferreira Ênio , Salas Carlos Edmundo , Lopes Miriam Teresa Paz
Published: February 2020

The proteolytic fraction (P1G10) from Vasconcellea cundinamarcensis, displays gastric protective and healing activities in different skin lesions in mice and human. In an excisional model, this fraction accelerates resolution of lesions and modulates inflammatory mediators. Based on these data, we assessed its anti-inflammatory activity in murine colitis model, induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS) adopted by its physiopathological similarity with human colitis. Twenty four hours after colitis induction followed by three days of treatment, P1G10 at 0.3 and 3.0 mg/Kg induced 30% increase in body weight (p < 0.0001) and ~80% reduction in colon macroscopic damage score (p < 0.05) compared to the untreated TNBS-induced colitis group. Histological analyses showed that 0.3 mg/Kg P1G10 reduced the inflammatory profile and tissue damage (47%, p < 0.05) when it was proteolytically active. Compared to TNBS group, 0.3 mg/Kg P1G10 reduced MPO activity (80%, p < 0.01), MCP-1 (47%, p < 0.05) and TNF-α (50%, no significant) and increased IL-10 (330%, p < 0.001) levels in the supernatant of colonic tissue homogenate. P1G10 treatment also reduced COX-2 expression (60%, p < 0.05) and metalloprotease-2 activity (39%, p < 0.05) while increased globet cell density (140%, p < 0.01), that contributes to mucus layer protection in colonic tissue. Taken together, these findings suggest that low doses of active P1G10 promotes lesion resolution, at least in part by its anti-inflammatory activity, in TNBS-colitis model.
Controlled Peptide-Mediated Vesicle Fusion Assessed by Simultaneous Dual-Colour Time-Lapsed Fluorescence Microscopy
Authors: Mora Nestor Lopez , Boyle Aimee L. , Kolck Bart Jan van , Rossen Anouk , Pokorná Šárka , Koukalová Alena , Šachl Radek , Hof Martin , Kros Alexander
Published: February 2020

We have employed a model system, inspired by SNARE proteins, to facilitate membrane fusion between Giant Unilamellar Vesicles (GUVs) and Large Unilamellar Vesicles (LUVs) under physiological conditions. In this system, two synthetic lipopeptide constructs comprising the coiled-coil heterodimer-forming peptides K4, (KIAALKE)4, or E4, (EIAALEK)4, a PEG spacer of variable length, and a cholesterol moiety to anchor the peptides into the liposome membrane replace the natural SNARE proteins. GUVs are functionalized with one of the lipopeptide constructs and the fusion process is triggered by adding LUVs bearing the complementary lipopeptide. Dual-colour time lapse fluorescence microscopy was used to visualize lipid- and content-mixing. Using conventional confocal microscopy, lipid mixing was observed on the lipid bilayer of individual GUVs. In addition to lipid-mixing, content-mixing assays showed a low efficiency due to clustering of K4-functionalized LUVs on the GUVs target membranes. We showed that, through the use of the non-ionic surfactant Tween 20, content-mixing between GUVs and LUVs could be improved, meaning this system has the potential to be employed for drug delivery in biological systems.
An endocannabinoid catabolic enzyme FAAH and its paralogs in an early land plant reveal evolutionary and functional relationship with eukaryotic orthologs
Authors: Haq Imdadul , Kilaru Aruna
Published: February 2020

Endocannabinoids were known to exist only among Animalia but recent report of their occurrence in early land plants prompted us to study its function and metabolism. In mammals, anandamide, as an endocannabinoid ligand, mediates several neurological and physiological processes, which are terminated by fatty acid amide hydrolase (FAAH). We identified nine orthologs of FAAH in the moss Physcomitrella patens (PpFAAH1 to PpFAAH9) with amidase signature and catalytic triad. The optimal amidase activity for PpFAAH1 was at 37 °C and pH 8.0, with higher specificity to anandamide. Further, the phylogeny and predicted structural analyses of the nine paralogs revealed that PpFAAH1 to PpFAAH4 were closely related to plant FAAH while PpFAAH6 to PpFAAH9 were to the rat FAAH, categorized based on the membrane binding cap, membrane access channel and substrate binding pocket. We also identified that a true ‘dynamic paddle’ that is responsible for tighter regulation of FAAH is recent in vertebrates and absent or not fully emerged in plants and non-vertebrates. These data reveal evolutionary and functional relationship among eukaryotic FAAH orthologs and features that contribute to versatility and tighter regulation of FAAH. Future studies will utilize FAAH mutants of moss to elucidate the role of anandamide in early land plants.
A Novel Mono-surface Antisymmetric 8Tx/16Rx Coil Array for Parallel Transmit Cardiac MRI in Pigs at 7T
Authors: Elabyad Ibrahim A. , Terekhov Maxim , Lohr David , Stefanescu Maria R. , Baltes Steffen , Schreiber Laura M.
Published: February 2020

A novel mono-surface antisymmetric 16-element transmit/receive (Tx/Rx) coil array was designed, simulated, constructed, and tested for cardiac magnetic resonance imaging (cMRI) in pigs at 7 T. The cardiac array comprised of a mono-surface 16-loops with two central elements arranged anti-symmetrically and flanked by seven elements on either side. The array was configured for parallel transmit (pTx) mode to have an eight channel transmit and 16-channel receive (8Tx/16Rx) coil array. Electromagnetic (EM) simulations, bench-top measurements, phantom, and MRI experiments with two pig cadavers (68 and 46 kg) were performed. Finally, the coil was used in pilot in-vivo measurements with a 60 kg pig. Flip angle (FA), geometry factor (g-factor), signal-to-noise ratio (SNR) maps, and high-resolution cardiac images were acquired with an in-plane resolution of 0.6 mm × 0.6 mm (in-vivo) and 0.3 mm × 0.3 mm (ex-vivo). The mean g-factor over the heart was 1.26 (R = 6). Static phase $${B}_{1}^{+}$$

B

1

+

shimming in a pig body phantom with the optimal phase vectors makes possible to improve the $${B}_{1}^{+}$$

B

1

+

homogeneity by factor > 2 and transmit efficiency by factor > 3 compared to zero phases (before RF shimming). Parallel imaging performed in the in-vivo measurements demonstrated well preserved diagnostic quality of the resulting images at acceleration factors up to R = 6. The described hardware design can be adapted for arrays optimized for animals and humans with a larger number of elements (32–64) while maintaining good decoupling for various MRI applications at UHF (e.g., cardiac, head, and spine).
Characterization of cereblon-dependent targeted protein degrader by visualizing the spatiotemporal ternary complex formation in cells
Authors: Kaji Tomohiro , Koga Hiroshi , Kuroha Mutsumi , Akimoto Toshihiko , Hayata Kenji
Published: February 2020

Targeted protein degradation (TPD) through a proteasome-dependent pathway induced by heterofunctional small molecules is initiated by the formation of a ternary complex with recruited E3 ligases. This complex formation affects the degradation ability of TPD molecules, and thus we tested for visualization of the intracellular dynamics of ternary complex formation. In this study, we applied the fluorescent-based technology detecting protein-protein interaction (Fluoppi) system, in which detectable fluorescent foci are formed when ternary complex formation induced by TPD molecules occurs in cells. We show here that cells coexpressing BRD4 and cereblon (CRBN) tagged with the Fluoppi system formed detectable foci in both live and fixed cells only when treated with BRD4-targeting degraders utilizing CRBN as an E3 ligase in dose- and time-dependent manners. Notably, the maintenance and efficacy of TPD molecule-induced foci formation correlated with the ability to degrade target proteins. Furthermore, we demonstrated that BRD4-targeting and FKBP12F36V-targeting degraders formed ternary complexes mainly in the nucleus and cytoplasm, respectively, suggesting that TPD molecules utilize the proteasome to degrade target proteins in their corresponding localized region. Our results also suggest that the Fluoppi system is a powerful tool for characterizing TPD molecules by visualizing the spatiotemporal formation of ternary complex.
Defining the risk of first intravenous immunoglobulin unresponsiveness in non-Asian patients with Kawasaki disease
Authors: Piram Maryam , Darce Bello Martha , Tellier Stéphanie , Di Filippo Sylvie , Boralevi Franck , Madhi Fouad , Meinzer Ulrich , Cimaz Rolando , Piedvache Celine , Koné-Paut Isabelle
Published: February 2020

About 10–20% of patients with Kawasaki disease (KD) are unresponsive to intravenous immunoglobulin (IVIg) and are at increased risk of coronary artery abnormalities (CAAs). Early identification is critical to initiate aggressive therapies, but available scoring systems lack sensitivity in non-Japanese populations. We investigated the accuracy of 3 Japanese scoring systems and studied factors associated with IVIg unresponsiveness in a large multiethnic French population of children with KD to build a new scoring system. Children admitted for KD between 2011–2014 in 65 centers were enrolled. Factors associated with second line-treatment; i.e. unresponsiveness to initial IVIg treatment, were analyzed by multivariate regression analysis. The performance of our score and the Kobayashi, Egami and Sano scores were compared in our population and in ethnic subgroups. Overall, 465 children were reported by 84 physicians; 425 were classified with KD (55% European Caucasian, 12% North African/Middle Eastern, 10% African/Afro-Caribbean, 3% Asian and 11% mixed). Eighty patients (23%) needed second-line treatment. Japanese scores had poor performance in our whole population (sensitivity 14–61%). On multivariate regression analysis, predictors of secondary treatment after initial IVIG were hepatomegaly, ALT level ≥30 IU/L, lymphocyte count <2400/mm3 and time to treatment <5 days. The best sensitivity (77%) and specificity (60%) of this model was with 1 point per variable and cut-off ≥2 points. The sensitivity remained good in our 3 main ethnic subgroups (74–88%). We identified predictors of IVIg resistance and built a new score with good sensitivity and acceptable specificity in a non-Asian population.
Immune correlates of postexposure vaccine protection against Marburg virus
Authors: Woolsey Courtney , Jankeel Allen , Matassov Demetrius , Geisbert Joan B. , Agans Krystle N. , Borisevich Viktoriya , Cross Robert W. , Deer Daniel J. , Fenton Karla A. , Latham Theresa E. , Gerardi Cheryl S. , Mire Chad E. , Eldridge John H. , Messaoudi Ilhem , Geisbert Thomas W.
Published: February 2020

Postexposure immunization can prevent disease and reduce transmission following pathogen exposure. The rapid immunostimulatory properties of recombinant vesicular stomatitis virus (rVSV)-based vaccines make them suitable postexposure treatments against the filoviruses Ebola virus and Marburg virus (MARV); however, the mechanisms that drive this protection are undefined. Previously, we reported 60–75% survival of rhesus macaques treated with rVSV vectors expressing MARV glycoprotein (GP) 20–30 minutes after a low dose exposure to the most pathogenic variant of MARV, Angola. Survival in this model was linked to production of GP-specific antibodies and lower viral load. To confirm these results and potentially identify novel correlates of postexposure protection, we performed a similar experiment, but analyzed plasma cytokine levels, frequencies of immune cell subsets, and the transcriptional response to infection in peripheral blood. In surviving macaques (80–89%), we observed induction of genes mapping to antiviral and interferon-related pathways early after treatment and a higher percentage of T helper 1 (Th1) and NK cells. In contrast, the response of non-surviving macaques was characterized by hypercytokinemia; a T helper 2 signature; recruitment of low HLA-DR expressing monocytes and regulatory T-cells; and transcription of immune checkpoint (e.g., PD-1, LAG3) genes. These results suggest dysregulated immunoregulation is associated with poor prognosis, whereas early innate signaling and Th1-skewed immunity are important for survival.
A salivary metabolite signature that reflects gingival host-microbe interactions: instability predicts gingivitis susceptibility
Authors: Fernandez-Gutierrez Marcela M. , Imangaliyev Sultan , Prodan Andrei , Loos Bruno G. , Keijser Bart J. F. , Kleerebezem Michiel
Published: February 2020

Several proteins and peptides in saliva were shown to stimulate gingival wound repair, but the role of salivary metabolites in this process remains unexplored. In vitro gingival re-epithelialization kinetics were determined using unstimulated saliva samples from healthy individuals collected during an experimental gingivitis study. Elastic net regression with stability selection identified a specific metabolite signature in a training dataset that was associated with the observed re-epithelialization kinetics and enabled its prediction for all saliva samples obtained in the clinical study. This signature encompassed ten metabolites, including plasmalogens, diacylglycerol and amino acid derivatives, which reflect enhanced host-microbe interactions. This association is in agreement with the positive correlation of the metabolite signature with the individual’s gingival bleeding index. Remarkably, intra-individual signature-variation over time was associated with elevated risk for gingivitis development. Unravelling how these metabolites stimulate wound repair could provide novel avenues towards therapeutic approaches in patients with impaired wound healing capacity.
Hyperproliferation is the main driver of metabolomic changes in psoriasis lesional skin
Authors: Pohla Liis , Ottas Aigar , Kaldvee Bret , Abram Kristi , Soomets Ursel , Zilmer Mihkel , Reemann Paula , Jaks Viljar , Kingo Külli
Published: February 2020

Systematic understanding of the metabolite signature of diseases may lead to a closer understanding of the disease pathogenesis and ultimately to the development of novel therapies and diagnostic tools. Here we compared for the first time the full metabolomic profiles of lesional and non-lesional skin biopsies obtained from plaque psoriasis patients and skin samples of healthy controls. Significant differences in the concentration levels of 29 metabolites were identified that provide several novel insights into the metabolic pathways of psoriatic lesions. The metabolomic profile of the lesional psoriatic skin is mainly characterized by hallmarks of increased cell proliferation. As no significant differences were identified between non-lesional skin and healthy controls we conclude that local inflammatory process that drives the increased cell proliferation is the main cause of the identified metabolomic shifts.
A semi-automated technique for adenoma quantification in the ApcMin mouse using FeatureCounter
Authors: Shepherd Amy L. , Smith A. Alexander T. , Wakelin Kirsty A. , Kuhn Sabine , Yang Jianping , Eccles David A. , Ronchese Franca
Published: February 2020

Colorectal cancer is a major contributor to death and disease worldwide. The ApcMin mouse is a widely used model of intestinal neoplasia, as it carries a mutation also found in human colorectal cancers. However, the method most commonly used to quantify tumour burden in these mice is manual adenoma counting, which is time consuming and poorly suited to standardization across different laboratories. We describe a method to produce suitable photographs of the small intestine of ApcMin mice, process them with an ImageJ macro, FeatureCounter, which automatically locates image features potentially corresponding to adenomas, and a machine learning pipeline to identify and quantify them. Compared to a manual method, the specificity (or True Negative Rate, TNR) and sensitivity (or True Positive Rate, TPR) of this method in detecting adenomas are similarly high at about 80% and 87%, respectively. Importantly, total adenoma area measures derived from the automatically-called tumours were just as capable of distinguishing high-burden from low-burden mice as those established manually. Overall, our strategy is quicker, helps control experimenter bias, and yields a greater wealth of information about each tumour, thus providing a convenient route to getting consistent and reliable results from a study.
Color-specific porosity in double pigmented natural 3d-nanoarchitectures of blue crab shell
Authors: Nekvapil Fran , Pinzaru Simona Cintă , Barbu–Tudoran Lucian , Suciu Maria , Glamuzina Branko , Tamaș Tudor , Chiș Vasile
Published: February 2020

3D-engineered nano-architectures with various functionalities are still difficult to obtain and translate for real-world applications. However, such nanomaterials are naturally abundant and yet wasted, but could trigger huge interest for blue bioeconomy, provided that our understanding of their ultrastructure-function is achieved. To date, the Bouligand pattern in crustaceans shell structure is believed to be unique. Here we demonstrated that in blue crab Callinectes sapidus, the 3D-nanoarchitecture is color-specific, while the blue and red-orange pigments interplay in different nano-sized channels and pores. Thinnest pores of about 20  nm are found in blue shell. Additionally, the blue pigment co-existence in specific Bouligand structure is proved for the green crab Carcinus aestuarii, although the crab does not appear blue. The pigments interplay, simultaneously detected by Raman spectroscopy in color-specific native cuticles, overturns our understanding in crustaceans coloration and may trigger the selective use of particular colored natural nanoarchitectures for broaden area of applications.
Local transplantation of adipose-derived stem cells has a significant therapeutic effect in a mouse model of rheumatoid arthritis
Authors: Ueyama Hideki , Okano Tadashi , Orita Kumi , Mamoto Kenji , Ii Masaaki , Sobajima Satoshi , Iwaguro Hideki , Nakamura Hiroaki
Published: February 2020

Adipose-derived stem cells (ADSCs) have anti-inflammatory and regenerative properties. The purpose of this study was to investigate the effect of locally administered ADSCs in a rheumatoid arthritis (RA) mouse model. In an in vivo experiment, single-cell ADSCs and three dimensionally-cultured ADSC spheroids were injected intra-articularly into the knees of RA model mice and histologically assessed. Marked improvement of synovial inflammation and articular cartilage regeneration was found in ADSC-treated mice. Proliferation, migration, and apoptosis assays of synovial fibroblasts incubated with single-cell and spheroid ADSCs were performed. The expression levels of total cytokine RNA in ADSC single cells, spheroids, and ADSC-treated inflammatory synovial fibroblasts were also evaluated by quantitative reverse transcription PCR. ADSCs suppressed the proliferation and migration of activated inflammatory cells and downregulated inflammatory cytokines. TSG-6 and TGFβ1 were significantly upregulated in ADSCs compared to controls and TGFβ1 was significantly upregulated in ADSC spheroids compared to single cells. The apoptosis rate of ADSC spheroids was significantly lower than that of single-cell ADSCs. These results indicated that intra-articular administration of ADSC single cells and spheroids was effective in an RA mouse model, offering a novel approach for the development of effective localized treatments for patients with RA.
Diagnostic ability of vessel density measured by spectral-domain optical coherence tomography angiography for glaucoma in patients with high myopia
Authors: Lee Kwanghyun , Maeng Kyung Joo , Kim Joo Yeon , Yang Heon , Choi Wungrak , Lee Sang Yeop , Seong Gong Je , Kim Chan Yun , Bae Hyoung Won
Published: February 2020

Although early glaucoma detection is important to prevent visual loss due to disease progression, its clinical diagnosis in highly myopic eyes is still difficult. Many studies using optical coherence tomography (OCT) angiography (OCTA) reported decreased vessel density (VD) in glaucomatous eyes compared to normal eyes. We evaluated the diagnostic ability of peripapillary VD and macular VD measured by OCTA, comparing them with conventional valuables such as peripapillary retinal nerve fibre layer (RNFL) thickness and macular ganglion cell-inner plexiform layer (GCIPL) thickness measured by OCT. We also calculated the average VD ratio (VDR) (average outer macular VD/average inner macular VD), superior VDR (superior outer macular VD/average inner macular VD), and inferior VDR (inferior outer macular VD/average inner macular VD). Totally, 169 eyes from 169 subjects were enrolled. Among OCTA measurements, the best diagnostic parameters were average VDR (AUROC: 0.852 and 0.909) and inferior VDR (AUROC: 0.820 and 0.941) in nonhighly and highly myopic eyes, respectively. Inferior VDR showed better diagnostic ability than most of the other OCT measurements including peripapillary RNFL thickness and macular GCIPL thickness in highly myopic eyes. Accordingly, OCTA measurements can be useful for diagnosing glaucoma in highly myopic eyes, especially when using calculated indices such as average VDR or inferior VDR.
Antifreeze protein complements cryoprotective dehydration in the freeze-avoiding springtail Megaphorura arctica
Authors: Graham Laurie A. , Boddington Marie E. , Holmstrup Martin , Davies Peter L.
Published: February 2020

The springtail, Megaphorura arctica, is freeze-avoiding and survives sub-zero temperatures by cryoprotective dehydration. At the onset of dehydration there is some supercooling of body fluids, and the danger of inoculative freezing, which would be lethal. To see if the springtails are protected by antifreeze proteins in this pre-equilibrium phase, we examined extracts from cold-acclimated M. arctica and recorded over 3 °C of freezing point depression. Proteins responsible for this antifreeze activity were isolated by ice affinity. They comprise isoforms ranging from 6.5 to 16.9 kDa, with an amino acid composition dominated by glycine (>35 mol%). Tryptic peptide sequences were used to identify the mRNA sequence coding for the smallest isoform. This antifreeze protein sequence has high similarity to one characterized in Hypogastrura harveyi, from a different springtail order. If these two antifreeze proteins are true homologs, we suggest their origin dates back to the Permian glaciations some 300 million years ago.