bluuebunny/crossref_metadata_2025 huggingface dataset:
A randomized placebo-controlled PET study of ketamine´s effect on serotonin1B receptor binding in patients with SSRI-resistant depression
Authors: Tiger Mikael , Veldman Emma R. , Ekman Carl-Johan , Halldin Christer , Svenningsson Per , Lundberg Johan
Published: June 2020

The glutamate N-methyl-d-aspartate receptor antagonist ketamine has a rapid antidepressant effect. Despite large research efforts, ketamine’s mechanism of action in major depressive disorder (MDD) has still not been determined. In rodents, the antidepressant properties of ketamine were found to be dependent on both the α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and the serotonin (5-HT)1B receptor. Low 5-HT1B receptor binding in limbic brain regions is a replicated finding in MDD. In non-human primates, AMPA-dependent increase in 5-HT1B receptor binding in the ventral striatum (VST) has been demonstrated after ketamine infusion. Thirty selective serotonin reuptake inhibitor-resistant MDD patients were recruited via advertisement and randomized to double-blind monotherapy with 0.5 mg/kg ketamine or placebo infusion. The patients were examined with the 5-HT1B receptor selective radioligand [11C]AZ10419369 and positron emission tomography (PET) before and 24–72 h after treatment. 5-HT1B receptor binding did not significantly alter in patients treated with ketamine compared with placebo. An increase in 5-HT1B receptor binding with 16.7 % (p = 0.036) was found in the hippocampus after one ketamine treatment. 5-HT1B receptor binding in VST at baseline correlated with MDD symptom ratings (r = −0.426, p = 0.019) and with reduction of depressive symptoms with ketamine (r = −0.644, p = 0.002). In conclusion, reduction of depressive symptoms in MDD patients after ketamine treatment is correlated inversely with baseline 5-HT1B receptor binding in VST. Further studies examining the role of 5-HT1B receptors in the antidepressant mechanism of action of ketamine should be conducted, homing in on the 5-HT1B receptor as an MDD treatment response marker.
Effects of early life exposure to traffic-related air pollution on brain development in juvenile Sprague-Dawley rats
Authors: Patten Kelley T. , González Eduardo A. , Valenzuela Anthony , Berg Elizabeth , Wallis Christopher , Garbow Joel R. , Silverman Jill L. , Bein Keith J. , Wexler Anthony S. , Lein Pamela J.
Published: June 2020

Epidemiological studies link traffic-related air pollution (TRAP) to increased risk for various neurodevelopmental disorders (NDDs); however, there are limited preclinical data demonstrating a causal relationship between TRAP and adverse neurodevelopmental outcomes. Moreover, much of the preclinical literature reports effects of concentrated ambient particles or diesel exhaust that do not recapitulate the complexity of real-world TRAP exposures. To assess the developmental neurotoxicity of more realistic TRAP exposures, we exposed male and female rats during gestation and early postnatal development to TRAP drawn directly from a traffic tunnel in Northern California and delivered to animals in real-time. We compared NDD-relevant neuropathological outcomes at postnatal days 51–55 in TRAP-exposed animals versus control subjects exposed to filtered air. As indicated by immunohistochemical analyses, TRAP significantly increased microglial infiltration in the CA1 hippocampus, but decreased astrogliosis in the dentate gyrus. TRAP exposure had no persistent effect on pro-inflammatory cytokine levels in the male or female brain, but did significantly elevate the anti-inflammatory cytokine IL-10 in females. In male rats, TRAP significantly increased hippocampal neurogenesis, while in females, TRAP increased granule cell layer width. TRAP had no effect on apoptosis in either sex. Magnetic resonance imaging revealed that TRAP-exposed females, but not males, also exhibited decreased lateral ventricular volume, which was correlated with increased granule cell layer width in the hippocampus in females. Collectively, these data indicate that exposure to real-world levels of TRAP during gestation and early postnatal development modulate neurodevelopment, corroborating epidemiological evidence of an association between TRAP exposure and increased risk of NDDs.
Olig2 SUMOylation protects against genotoxic damage response by antagonizing p53 gene targeting
Authors: Liu Huiqing , Weng Weiji , Guo Rongjun , Zhou Jie , Xue Jun , Zhong Shan , Cheng Jinke , Zhu Michael X. , Pan Si-Jian , Li Yong
Published: June 2020

Posttranslational modifications of nuclear proteins, including transcription factors, nuclear receptors, and their coregulators, have attracted much attention in cancer research. Although phosphorylation of oligodendrocyte transcription factor 2 (Olig2) may contribute to the notorious resistance of gliomas to radiation and genotoxic drugs, the precise mechanisms remain elusive. We show here that in addition to phosphorylation, Olig2 is also conjugated by small ubiquitin-like modifier-1 (SUMO1) at three lysine residues K27, K76, and K112. SUMOylation is required for Olig2 to suppress p53-mediated cell cycle arrest and apoptosis induced by genotoxic damage, and to enhance resistance to temozolomide (TMZ) in glioma. Both SUMOylation and triple serine motif (TSM) phosphorylation of Olig2 are required for the antiapoptotic function. Olig2 SUMOylation enhances its genetic targeting ability, which in turn occludes p53 recruitment to Cdkn1a promoter for DNA-damage responses. Our work uncovers a SUMOylation-dependent regulatory mechanism of Olig2 in regulating cancer survival.
CAMK2A supported tumor initiating cells of lung adenocarcinoma by upregulating SOX2 through EZH2 phosphorylation
Authors: Wang Si-Qi , Liu Jing , Qin Jing , Zhu Yun , Tin Vicky Pui-Chi , Yam Judy Wai Ping , Wong Maria Pik , Xiao Zhi-Jie
Published: June 2020

Tumor initiating cells (TIC) of lung cancer are mainly induced by stress-related plasticity. Calcium/Calmodulin dependent protein kinase II alpha (CAMK2A) is a key calcium signaling molecule activated by exogenous and endogenous stimuli with effects on multiple cell functions but little is known about its role on TIC. In human lung adenocarcinomas (AD), CAMK2A was aberrantly activated in a proportion of cases and was an independent risk factor predicting shorter survivals. Functionally, CAMK2A enhanced TIC phenotypes in vitro and in vivo. CAMK2A regulated SOX2 expression by reducing H3K27me3 and EZH2 occupancy at SOX2 regulatory regions, leading to its epigenetic de-repression with functional consequences. Further, CAMK2A caused kinase-dependent phosphorylation of EZH2 at T487 with suppression of EZH2 activity. Together, the data demonstrated the CAMK2A-EZH2-SOX2 axis on TIC regulation. This study provided phenotypic and mechanistic evidence for the TIC supportive role of CAMK2A, implicating a novel predictive and therapeutic target for lung cancer.
Small extracellular vesicles derived from human MSCs prevent allergic airway inflammation via immunomodulation on pulmonary macrophages
Authors: Fang Shu-Bin , Zhang Hong-Yu , Meng Xiang-Ci , Wang Cong , He Bi-Xin , Peng Ya-Qi , Xu Zhi-Bin , Fan Xing-Liang , Wu Zhang-Jin , Wu Zi-Cong , Zheng Song-Guo , Fu Qing-Ling
Published: June 2020

Allergic airway inflammation is a major public health disease that affects up to 300 million people in the world. However, its management remains largely unsatisfactory. The dysfunction of pulmonary macrophages contributes greatly to the development of allergic airway inflammation. It has been reported that small extracellular vesicles derived from mesenchymal stromal cells (MSC-sEV) were able to display extensive therapeutic effects in some immune diseases. This study aimed to investigate the effects of MSC-sEV on allergic airway inflammation, and the role of macrophages involved in it. We successfully isolated MSC-sEV by using anion exchange chromatography, which were morphologically intact and positive for the specific EV markers. MSC-sEV significantly reduced infiltration of inflammatory cells and number of epithelial goblet cells in lung tissues of mice with allergic airway inflammation. Levels of inflammatory cells and cytokines in bronchoalveolar lavage fluid were also significantly decreased. Importantly, levels of monocytes-derived alveolar macrophages and M2 macrophages were significantly reduced by MSC-sEV. MSC-sEV were excreted through spleen and liver at 24 h post-administration in mice, and were able to be taken in by macrophages both in vivo and in vitro. In addition, proteomics analysis of MSC-sEV revealed that the indicated three types of MSC-sEV contained different quantities of proteins and shared 312 common proteins, which may be involved in the therapeutic effects of MSC-sEV. In total, our study demonstrated that MSC-sEV isolated by anion exchange chromatography were able to ameliorate Th2-dominant allergic airway inflammation through immunoregulation on pulmonary macrophages, suggesting that MSC-sEV were promising alternative therapy for allergic airway inflammation in the future.
Functional reconstruction of injured corpus cavernosa using 3D-printed hydrogel scaffolds seeded with HIF-1α-expressing stem cells
Authors: An Geng , Guo Feixiang , Liu Xuemin , Wang Zhifang , Zhu Ye , Fan Yong , Xuan Chengkai , Li Yan , Wu Hongkai , Shi Xuetao , Mao Chuanbin
Published: June 2020

Injury of corpus cavernosa results in erectile dysfunction, but its treatment has been very difficult. Here we construct heparin-coated 3D-printed hydrogel scaffolds seeded with hypoxia inducible factor-1α (HIF-1α)-mutated muscle-derived stem cells (MDSCs) to develop bioengineered vascularized corpora. HIF-1α-mutated MDSCs significantly secrete various angiogenic factors in MDSCs regardless of hypoxia or normoxia. The biodegradable scaffolds, along with MDSCs, are implanted into corpus cavernosa defects in a rabbit model to show good histocompatibility with no immunological rejection, support vascularized tissue ingrowth, and promote neovascularisation to repair the defects. Evaluation of morphology, intracavernosal pressure, elasticity and shrinkage of repaired cavernous tissue prove that the bioengineered corpora scaffolds repair the defects and recover penile erectile and ejaculation function successfully. The function recovery restores the reproductive capability of the injured male rabbits. Our work demonstrates that the 3D-printed hydrogels with angiogenic cells hold great promise for penile reconstruction to restore reproductive capability of males.
Integrating whole-genome sequencing within the National Antimicrobial Resistance Surveillance Program in the Philippines
Authors: Argimón Silvia , Masim Melissa A. L. , Gayeta June M. , Lagrada Marietta L. , Macaranas Polle K. V. , Cohen Victoria , Limas Marilyn T. , Espiritu Holly O. , Palarca Janziel C. , Chilam Jeremiah , Jamoralin Manuel C. , Villamin Alfred S. , Borlasa Janice B. , Olorosa Agnettah M. , Hernandez Lara F. T. , Boehme Karis D. , Jeffrey Benjamin , Abudahab Khalil , Hufano Charmian M. , Sia Sonia B. , Stelling John , Holden Matthew T. G. , Aanensen David M. , Carlos Celia C.
Published: June 2020

National networks of laboratory-based surveillance of antimicrobial resistance (AMR) monitor resistance trends and disseminate these data to AMR stakeholders. Whole-genome sequencing (WGS) can support surveillance by pinpointing resistance mechanisms and uncovering transmission patterns. However, genomic surveillance is rare in low- and middle-income countries. Here, we implement WGS within the established Antimicrobial Resistance Surveillance Program of the Philippines via a binational collaboration. In parallel, we characterize bacterial populations of key bug-drug combinations via a retrospective sequencing survey. By linking the resistance phenotypes to genomic data, we reveal the interplay of genetic lineages (strains), AMR mechanisms, and AMR vehicles underlying the expansion of specific resistance phenotypes that coincide with the growing carbapenem resistance rates observed since 2010. Our results enhance our understanding of the drivers of carbapenem resistance in the Philippines, while also serving as the genetic background to contextualize ongoing local prospective surveillance.
Inactivation of Arid1a in the endometrium is associated with endometrioid tumorigenesis through transcriptional reprogramming
Authors: Suryo Rahmanto Yohan , Shen Wenjing , Shi Xu , Chen Xi , Yu Yu , Yu Zheng-Cheng , Miyamoto Tsutomu , Lee Meng-Horng , Singh Vivek , Asaka Ryoichi , Shimberg Geoffrey , Vitolo Michele I. , Martin Stuart S. , Wirtz Denis , Drapkin Ronny , Xuan Jianhua , Wang Tian-Li , Shih Ie-Ming
Published: June 2020

Somatic inactivating mutations of ARID1A, a SWI/SNF chromatin remodeling gene, are prevalent in human endometrium-related malignancies. To elucidate the mechanisms underlying how ARID1A deleterious mutation contributes to tumorigenesis, we establish genetically engineered murine models with Arid1a and/or Pten conditional deletion in the endometrium. Transcriptomic analyses on endometrial cancers and precursors derived from these mouse models show a close resemblance to human uterine endometrioid carcinomas. We identify transcriptional networks that are controlled by Arid1a and have an impact on endometrial tumor development. To verify findings from the murine models, we analyze ARID1AWT and ARID1AKO human endometrial epithelial cells. Using a system biology approach and functional studies, we demonstrate that ARID1A-deficiency lead to loss of TGF-β tumor suppressive function and that inactivation of ARID1A/TGF-β axis promotes migration and invasion of PTEN-deleted endometrial tumor cells. These findings provide molecular insights into how ARID1A inactivation accelerates endometrial tumor progression and dissemination, the major causes of cancer mortality.
Designing antifilarial drug trials using clinical trial simulators
Authors: Walker Martin , Hamley Jonathan I. D. , Milton Philip , Monnot Frédéric , Pedrique Belén , Basáñez Maria-Gloria
Published: June 2020

Lymphatic filariasis and onchocerciasis are neglected tropical diseases (NTDs) targeted for elimination by mass (antifilarial) drug administration. These drugs are predominantly active against the microfilarial progeny of adult worms. New drugs or combinations are needed to improve patient therapy and to enhance the effectiveness of interventions in persistent hotspots of transmission. Several therapies and regimens are currently in (pre-)clinical testing. Clinical trial simulators (CTSs) project patient outcomes to inform the design of clinical trials but have not been widely applied to NTDs, where their resource-saving payoffs could be highly beneficial. We demonstrate the utility of CTSs using our individual-based onchocerciasis transmission model (EPIONCHO-IBM) that projects trial outcomes of a hypothetical macrofilaricidal drug. We identify key design decisions that influence the power of clinical trials, including participant eligibility criteria and post-treatment follow-up times for measuring infection indicators. We discuss how CTSs help to inform target product profiles.
An evolutionary path to altered cofactor specificity in a metalloenzyme
Authors: Barwinska-Sendra Anna , Garcia Yuritzi M. , Sendra Kacper M. , Baslé Arnaud , Mackenzie Eilidh S. , Tarrant Emma , Card Patrick , Tabares Leandro C. , Bicep Cédric , Un Sun , Kehl-Fie Thomas E. , Waldron Kevin J.
Published: June 2020

Almost half of all enzymes utilize a metal cofactor. However, the features that dictate the metal utilized by metalloenzymes are poorly understood, limiting our ability to manipulate these enzymes for industrial and health-associated applications. The ubiquitous iron/manganese superoxide dismutase (SOD) family exemplifies this deficit, as the specific metal used by any family member cannot be predicted. Biochemical, structural and paramagnetic analysis of two evolutionarily related SODs with different metal specificity produced by the pathogenic bacterium Staphylococcus aureus identifies two positions that control metal specificity. These residues make no direct contacts with the metal-coordinating ligands but control the metal’s redox properties, demonstrating that subtle architectural changes can dramatically alter metal utilization. Introducing these mutations into S. aureus alters the ability of the bacterium to resist superoxide stress when metal starved by the host, revealing that small changes in metal-dependent activity can drive the evolution of metalloenzymes with new cofactor specificity.
A genome-wide gain-of-function screen identifies CDKN2C as a HBV host factor
Authors: Eller Carla , Heydmann Laura , Colpitts Che C. , El Saghire Houssein , Piccioni Federica , Jühling Frank , Majzoub Karim , Pons Caroline , Bach Charlotte , Lucifora Julie , Lupberger Joachim , Nassal Michael , Cowley Glenn S. , Fujiwara Naoto , Hsieh Sen-Yung , Hoshida Yujin , Felli Emanuele , Pessaux Patrick , Sureau Camille , Schuster Catherine , Root David E. , Verrier Eloi R. , Baumert Thomas F.
Published: June 2020

Chronic HBV infection is a major cause of liver disease and cancer worldwide. Approaches for cure are lacking, and the knowledge of virus-host interactions is still limited. Here, we perform a genome-wide gain-of-function screen using a poorly permissive hepatoma cell line to uncover host factors enhancing HBV infection. Validation studies in primary human hepatocytes identified CDKN2C as an important host factor for HBV replication. CDKN2C is overexpressed in highly permissive cells and HBV-infected patients. Mechanistic studies show a role for CDKN2C in inducing cell cycle G1 arrest through inhibition of CDK4/6 associated with the upregulation of HBV transcription enhancers. A correlation between CDKN2C expression and disease progression in HBV-infected patients suggests a role in HBV-induced liver disease. Taken together, we identify a previously undiscovered clinically relevant HBV host factor, allowing the development of improved infectious model systems for drug discovery and the study of the HBV life cycle.
Site-selective electrooxidation of methylarenes to aromatic acetals
Authors: Xiong Peng , Zhao Huai-Bo , Fan Xue-Ting , Jie Liang-Hua , Long Hao , Xu Pin , Liu Zhan-Jiang , Wu Zheng-Jian , Cheng Jun , Xu Hai-Chao
Published: June 2020

Aldehyde is one of most synthetically versatile functional groups and can participate in numerous chemical transformations. While a variety of simple aromatic aldehydes are commercially available, those with a more complex substitution pattern are often difficult to obtain. Benzylic oxygenation of methylarenes is a highly attractive method for aldehyde synthesis as the starting materials are easy to obtain and handle. However, regioselective oxidation of functionalized methylarenes, especially those that contain heterocyclic moieties, to aromatic aldehydes remains a significant challenge. Here we show an efficient electrochemical method that achieves site-selective electrooxidation of methyl benzoheterocycles to aromatic acetals without using chemical oxidants or transition-metal catalysts. The acetals can be converted to the corresponding aldehydes through hydrolysis in one-pot or in a separate step. The synthetic utility of our method is highlighted by its application to the efficient preparation of the antihypertensive drug telmisartan.
Model-based analysis of sample index hopping reveals its widespread artifacts in multiplexed single-cell RNA-sequencing
Authors: Farouni Rick , Djambazian Haig , Ferri Lorenzo E. , Ragoussis Jiannis , Najafabadi Hamed S.
Published: June 2020

Index hopping is the main cause of incorrect sample assignment of sequencing reads in multiplexed pooled libraries. We introduce a statistical model for estimating the sample index-hopping rate in multiplexed droplet-based single-cell RNA-seq data and for probabilistic inference of the true sample of origin of hopped reads. We analyze several datasets and estimate the sample index hopping probability to range between 0.003–0.009, a small number that counter-intuitively gives rise to a large fraction of phantom molecules — the fraction of phantom molecules exceeds 8% in more than 25% of samples and reaches as high as 85% in low-complexity samples. Phantom molecules lead to widespread complications in downstream analyses, including transcriptome mixing across cells, emergence of phantom copies of cells from other samples, and misclassification of empty droplets as cells. We demonstrate that our approach can correct for these artifacts by accurately purging the majority of phantom molecules from the data.
Ultrastrong coupling between nanoparticle plasmons and cavity photons at ambient conditions
Authors: Baranov Denis G. , Munkhbat Battulga , Zhukova Elena , Bisht Ankit , Canales Adriana , Rousseaux Benjamin , Johansson Göran , Antosiewicz Tomasz J. , Shegai Timur
Published: June 2020

Ultrastrong coupling is a distinct regime of electromagnetic interaction that enables a rich variety of intriguing physical phenomena. Traditionally, this regime has been reached by coupling intersubband transitions of multiple quantum wells, superconducting artificial atoms, or two-dimensional electron gases to microcavity resonators. However, employing these platforms requires demanding experimental conditions such as cryogenic temperatures, strong magnetic fields, and high vacuum. Here, we use a plasmonic nanorod array positioned at the antinode of a resonant optical Fabry-Pérot microcavity to reach the ultrastrong coupling (USC) regime at ambient conditions and without the use of magnetic fields. From optical measurements we extract the value of the interaction strength over the transition energy as high as g/ω ~ 0.55, deep in the USC regime, while the nanorod array occupies only ∼4% of the cavity volume. Moreover, by comparing the resonant energies of the coupled and uncoupled systems, we indirectly observe up to ∼10% modification of the ground-state energy, which is a hallmark of USC. Our results suggest that plasmon-microcavity polaritons are a promising platform for room-temperature USC realizations in the optical and infrared ranges, and may lead to the long-sought direct visualization of the vacuum energy modification.
Glycan-dependent cell adhesion mechanism of Tc toxins
Authors: Roderer Daniel , Bröcker Felix , Sitsel Oleg , Kaplonek Paulina , Leidreiter Franziska , Seeberger Peter H. , Raunser Stefan
Published: June 2020

Toxin complex (Tc) toxins are virulence factors of pathogenic bacteria. Tcs are composed of three subunits: TcA, TcB and TcC. TcA facilitates receptor–toxin interaction and membrane permeation, TcB and TcC form a toxin-encapsulating cocoon. While the mechanisms of holotoxin assembly and pore formation have been described, little is known about receptor binding of TcAs. Here, we identify heparins/heparan sulfates and Lewis antigens as receptors for different TcAs from insect and human pathogens. Glycan array screening reveals that all tested TcAs bind negatively charged heparins. Cryo-EM structures ofMorganella morganiiTcdA4 andXenorhabdus nematophilaXptA1 reveal that heparins/heparan sulfates unexpectedly bind to different regions of the shell domain, including receptor-binding domains. In addition,Photorhabdus luminescensTcdA1 binds to Lewis antigens with micromolar affinity. Here, the glycan interacts with the receptor-binding domain D of the toxin. Our results suggest a glycan dependent association mechanism of Tc toxins on the host cell surface.
Skeletal muscle enhancer interactions identify genes controlling whole-body metabolism
Authors: Williams Kristine , Ingerslev Lars R. , Bork-Jensen Jette , Wohlwend Martin , Hansen Ann Normann , Small Lewin , Ribel-Madsen Rasmus , Astrup Arne , Pedersen Oluf , Auwerx Johan , Workman Christopher T. , Grarup Niels , Hansen Torben , Barrès Romain
Published: June 2020

Obesity and type 2 diabetes (T2D) are metabolic disorders influenced by lifestyle and genetic factors that are characterized by insulin resistance in skeletal muscle, a prominent site of glucose disposal. Numerous genetic variants have been associated with obesity and T2D, of which the majority are located in non-coding DNA regions. This suggests that most variants mediate their effect by altering the activity of gene-regulatory elements, including enhancers. Here, we map skeletal muscle genomic enhancer elements that are dynamically regulated after exposure to the free fatty acid palmitate or the inflammatory cytokine TNFα. By overlapping enhancer positions with the location of disease-associated genetic variants, and resolving long-range chromatin interactions between enhancers and gene promoters, we identify target genes involved in metabolic dysfunction in skeletal muscle. The majority of these genes also associate with altered whole-body metabolic phenotypes in the murine BXD genetic reference population. Thus, our combined genomic investigations identified genes that are involved in skeletal muscle metabolism.
Suppression of unwanted CRISPR-Cas9 editing by co-administration of catalytically inactivating truncated guide RNAs
Authors: Rose John C. , Popp Nicholas A. , Richardson Christopher D. , Stephany Jason J. , Mathieu Julie , Wei Cindy T. , Corn Jacob E. , Maly Dustin J. , Fowler Douglas M.
Published: June 2020

CRISPR-Cas9 nucleases are powerful genome engineering tools, but unwanted cleavage at off-target and previously edited sites remains a major concern. Numerous strategies to reduce unwanted cleavage have been devised, but all are imperfect. Here, we report that off-target sites can be shielded from the active Cas9•single guide RNA (sgRNA) complex through the co-administration of dead-RNAs (dRNAs), truncated guide RNAs that direct Cas9 binding but not cleavage. dRNAs can effectively suppress a wide-range of off-targets with minimal optimization while preserving on-target editing, and they can be multiplexed to suppress several off-targets simultaneously. dRNAs can be combined with high-specificity Cas9 variants, which often do not eliminate all unwanted editing. Moreover, dRNAs can prevent cleavage of homology-directed repair (HDR)-corrected sites, facilitating scarless editing by eliminating the need for blocking mutations. Thus, we enable precise genome editing by establishing a flexible approach for suppressing unwanted editing of both off-targets and HDR-corrected sites.
Specific fibroblast subpopulations and neuronal structures provide local sources of Vegfc-processing components during zebrafish lymphangiogenesis
Authors: Wang Guangxia , Muhl Lars , Padberg Yvonne , Dupont Laura , Peterson-Maduro Josi , Stehling Martin , le Noble Ferdinand , Colige Alain , Betsholtz Christer , Schulte-Merker Stefan , van Impel Andreas
Published: June 2020

Proteolytical processing of the growth factor VEGFC through the concerted activity of CCBE1 and ADAMTS3 is required for lymphatic development to occur. How these factors act together in time and space, and which cell types produce these factors is not understood. Here we assess the function of Adamts3 and the related protease Adamts14 during zebrafish lymphangiogenesis and show both proteins to be able to process Vegfc. Only the simultaneous loss of both protein functions results in lymphatic defects identical tovegfcloss-of-function situations. Cell transplantation experiments demonstrate neuronal structures and/or fibroblasts to constitute cellular sources not only for both proteases but also for Ccbe1 and Vegfc. We further show that this locally restricted Vegfc maturation is needed to trigger normal lymphatic sprouting and directional migration. Our data provide a single-cell resolution model for establishing secretion and processing hubs for Vegfc during developmental lymphangiogenesis.
Tetrameric architecture of an active phenol-bound form of the AAA+ transcriptional regulator DmpR
Authors: Park Kwang-Hyun , Kim Sungchul , Lee Su-Jin , Cho Jee-Eun , Patil Vinod Vikas , Dumbrepatil Arti Baban , Song Hyung-Nam , Ahn Woo-Chan , Joo Chirlmin , Lee Seung-Goo , Shingler Victoria , Woo Eui-Jeon
Published: June 2020

The Pseudomonas putida phenol-responsive regulator DmpR is a bacterial enhancer binding protein (bEBP) from the AAA+ ATPase family. Even though it was discovered more than two decades ago and has been widely used for aromatic hydrocarbon sensing, the activation mechanism of DmpR has remained elusive. Here, we show that phenol-bound DmpR forms a tetramer composed of two head-to-head dimers in a head-to-tail arrangement. The DmpR-phenol complex exhibits altered conformations within the C-termini of the sensory domains and shows an asymmetric orientation and angle in its coiled-coil linkers. The structural changes within the phenol binding sites and the downstream ATPase domains suggest that the effector binding signal is propagated through the coiled-coil helixes. The tetrameric DmpR-phenol complex interacts with the σ54 subunit of RNA polymerase in presence of an ATP analogue, indicating that DmpR-like bEBPs tetramers utilize a mechanistic mode distinct from that of hexameric AAA+ ATPases to activate σ54-dependent transcription.
Immunotherapy with engineered bacteria by targeting the STING pathway for anti-tumor immunity
Authors: Leventhal Daniel S. , Sokolovska Anna , Li Ning , Plescia Christopher , Kolodziej Starsha A. , Gallant Carey W. , Christmas Rudy , Gao Jian-Rong , James Michael J. , Abin-Fuentes Andres , Momin Munira , Bergeron Christopher , Fisher Adam , Miller Paul F. , West Kip A. , Lora Jose M.
Published: June 2020

Synthetic biology is a powerful tool to create therapeutics which can be rationally designed to enable unique and combinatorial functionalities. Here we utilize non-pathogenic E coli Nissle as a versatile platform for the development of a living biotherapeutic for the treatment of cancer. The engineered bacterial strain, referred to as SYNB1891, targets STING-activation to phagocytic antigen-presenting cells (APCs) in the tumor and activates complementary innate immune pathways. SYNB1891 treatment results in efficacious antitumor immunity with the formation of immunological memory in murine tumor models and robust activation of human APCs. SYNB1891 is designed to meet manufacturability and regulatory requirements with built in biocontainment features which do not compromise its efficacy. This work provides a roadmap for the development of future therapeutics and demonstrates the transformative potential of synthetic biology for the treatment of human disease when drug development criteria are incorporated into the design process for a living medicine.
Two linear epitopes on the SARS-CoV-2 spike protein that elicit neutralising antibodies in COVID-19 patients
Authors: Poh Chek Meng , Carissimo Guillaume , Wang Bei , Amrun Siti Naqiah , Lee Cheryl Yi-Pin , Chee Rhonda Sin-Ling , Fong Siew-Wai , Yeo Nicholas Kim-Wah , Lee Wen-Hsin , Torres-Ruesta Anthony , Leo Yee-Sin , Chen Mark I-Cheng , Tan Seow-Yen , Chai Louis Yi Ann , Kalimuddin Shirin , Kheng Shirley Seah Gek , Thien Siew-Yee , Young Barnaby Edward , Lye David C. , Hanson Brendon John , Wang Cheng-I , Renia Laurent , Ng Lisa F. P.
Published: June 2020

Given the ongoing SARS-CoV-2 pandemic, identification of immunogenic targets against the coronavirus spike glycoprotein will provide crucial advances towards the development of sensitive diagnostic tools and potential vaccine candidate targets. In this study, using pools of overlapping linear B-cell peptides, we report two IgG immunodominant regions on SARS-CoV-2 spike glycoprotein that are recognised by sera from COVID-19 convalescent patients. Notably, one is specific to SARS-CoV-2, which is located in close proximity to the receptor binding domain. The other region, which is localised at the fusion peptide, could potentially function as a pan-SARS target. Functionally, antibody depletion assays demonstrate that antibodies targeting these immunodominant regions significantly alter virus neutralisation capacities. Taken together, identification and validation of these neutralising B-cell epitopes will provide insights towards the design of diagnostics and vaccine candidates against this high priority coronavirus.
A large-area AgNW-modified textile with high-performance electromagnetic interference shielding
Authors: Liu Junchen , Lin Sen , Huang Kai , Jia Chao , Wang Qingmin , Li Ziwei , Song Jianan , Liu Zhenglian , Wang Haiyang , Lei Ming , Wu Hui
Published: June 2020

Manufacturing a flexible, light, large-area, and high-efficiency electromagnetic shielding materials in a straightforward and cost-effective manner presently remains a significant challenge. In this work, we propose a conductive network design and verify its electromagnetic interference (EMI) shielding effectiveness (SE) by simulation. Using the structure and parameters obtained by simulation, we prepare a flexible EMI shielding material using silver nanowires (AgNWs)/polyvinyl butyral (PVB) ethanol solution and textile substructure via a facile immersing method. In the frequency range of 5–18 GHz, the AgNWs/PVB textile with 1.4 mm thickness achieves an EMI SE of 59 dB, which exceeds the requirements for commercial applications. Due to the low density of 56 mg/cm3, specific shielding effectiveness (SSE) of this material reaches 1053 dB m3/g. It is found that the AgNWs/PVB textile is more resistant to washing with water and oxidation than AgNWs textile without a PVB protector. As a result, the conductivity of AgNWs/PVB textile exhibits no change after washing with water and varies slightly after being kept in hot air. We find that a signal monitor is unable to detect a signal emitted by a mobile phone from a jacket lined with AgNWs/PVB textile. AgNWs/PVB textile with these properties can be mass-produced as high-efficiency EMI shielding material for commercial applications.
The Help for Hay Fever community pharmacy-based pilot randomised controlled trial for intermittent allergic rhinitis
Authors: Smith Sarah , Porteous Terry , Bond Christine , Francis Jill , Lee Amanda J. , Lowrie Richard , Scotland Graham , Sheikh Aziz , Thomas Mike , Wyke Sally , Smith Lorraine
Published: June 2020

Management of intermittent allergic rhinitis (IAR) is suboptimal in the UK. An Australian community pharmacy-based intervention has been shown to help patients better self-manage their IAR. We conducted a pilot cluster RCT in 12 Scottish community pharmacies to assess transferability of the Australian intervention. Trained staff in intervention pharmacies delivered the intervention to eligible customers (n = 60). Non-intervention pharmacy participants (n = 65) received usual care. Outcome measures included effect size of change in the mini-Rhinoconjunctivitis Quality of Life Questionnaire (miniRQLQ) between baseline, 1-week and 6-week follow-up. Trial procedures were well received by pharmacy staff, and customer satisfaction with the intervention was high. The standardised effect size for miniRQLQ total score was −0.46 (95% CI, −1.05, 0.13) for all participants and −0.14 (95% CI,−0.86, 0.57) in the complete case analysis, suggesting a small overall treatment effect in the intervention group. A full-scale RCT is warranted to fully evaluate the effectiveness of this service.
The sub-band structure of atomically sharp dopant profiles in silicon
Authors: Mazzola Federico , Chen Chin-Yi , Rahman Rajib , Zhu Xie-Gang , Polley Craig M. , Balasubramanian Thiagarajan , King Phil D. C. , Hofmann Philip , Miwa Jill A. , Wells Justin W.
Published: June 2020

The downscaling of silicon-based structures and proto-devices has now reached the single-atom scale, representing an important milestone for the development of a silicon-based quantum computer. One especially notable platform for atomic-scale device fabrication is the so-called Si:P δ-layer, consisting of an ultra-dense and sharp layer of dopants within a semiconductor host. Whilst several alternatives exist, it is on the Si:P platform that many quantum proto-devices have been successfully demonstrated. Motivated by this, both calculations and experiments have been dedicated to understanding the electronic structure of the Si:P δ-layer platform. In this work, we use high-resolution angle-resolved photoemission spectroscopy to reveal the structure of the electronic states which exist because of the high dopant density of the Si:P δ-layer. In contrast to published theoretical work, we resolve three distinct bands, the most occupied of which shows a large anisotropy and significant deviation from simple parabolic behaviour. We investigate the possible origins of this fine structure, and conclude that it is primarily a consequence of the dielectric constant being large (ca. double that of bulk Si). Incorporating this factor into tight-binding calculations leads to a major revision of band structure; specifically, the existence of a third band, the separation of the bands, and the departure from purely parabolic behaviour. This new understanding of the band structure has important implications for quantum proto-devices which are built on the Si:P δ-layer platform.
Emotion dysregulation mediates the relationship between nightmares and psychotic experiences: results from a student population
Authors: Akram Umair , Gardani Maria , Irvine Kamila , Allen Sarah , Ypsilanti Antonia , Lazuras Lambros , Drabble Jennifer , Stevenson Jodie C. , Akram Asha
Published: June 2020

Sleep disruption is commonly associated with psychotic experiences. While sparse, the literature to date highlights nightmares and related distress as prominent risk factors for psychosis in students. We aimed to further explore the relationship between specific nightmare symptoms and psychotic experiences in university students while examining the mediating role of emotion dysregulation. A sample (N = 1273) of student respondents from UK universities completed measures of psychotic experiences, nightmare disorder symptomology and emotion dysregulation. Psychotic experiences were significantly more prevalent in students reporting nightmares (n = 757) relative to those who did not (n = 516). Hierarchical linear regression analysis showed that psychotic experiences were significantly associated (Adjusted R2 = 32.4%) with perceived nightmare intensity, consequences and resulting awakenings, and with emotion regulation difficulties. Furthermore, multiple mediation analysis showed that the association between psychotic experiences and nightmare factors was mediated by emotion regulation difficulties. Adaptive regulation of dream content during rapid eye-movement sleep has previously been demonstrated to attenuate surges in affective arousal by controlling the intensity and variability of emotional content. Difficulties in emotion regulation may partially explain the experience of more intense and disruptive nightmares among individuals with psychotic experiences. Emotion regulation may represent an important control mechanism that safeguards dream content and sleep quality.
Novel biomarkers of a peripheral blood interferon signature associated with drug-naïve early arthritis patients distinguish persistent from self-limiting disease course
Authors: Seyhan Attila A. , Gregory Bernard , Cribbs Adam P. , Bhalara Sundeept , Li Yizheng , Loreth Christine , Zhang Ying , Guo Yongjing , Lin Lih-Ling , Feldmann Marc , Williams Lynn M. , Brennan Fionula M. , Taylor Peter C.
Published: June 2020

We profiled gene expression signatures to distinguish rheumatoid arthritis (RA) from non-inflammatory arthralgia (NIA), self-limiting arthritis (SLA), and undifferentiated arthritis (UA) as compared to healthy controls as novel potential biomarkers for therapeutic responsiveness. Global gene expression profiles of PBMCs from 43 drug-naïve patients presenting with joint symptoms were evaluated and differentially expressed genes identified by comparative analysis with 24 healthy volunteers. Patients were assessed at presentation with follow up at 6 and 12 months. Gene ontology and network pathway analysis were performed using DAVID Bioinformatics Resources v6.7. Gene expression profiles were also determined after disease-modifying anti-rheumatic drug (DMARD) treatment in the inflammatory arthritis groups (i.e. RA and UA) and confirmed by qRT-PCR. Receiver operating characteristic (ROC) curves analysis and Area Under the Curve (AUC) estimation were performed to assess the diagnostic value of candidate gene expression signatures. A type I interferon (IFN) gene signature distinguished DMARD-naïve patients who will subsequently develop persistent inflammatory arthritis (i.e. RA and UA) from those with NIA. In patients with RA, the IFN signature is characterised by up-regulation of SIGLEC1 (p = 0.00597) and MS4A4A (p = 0.00000904). We also identified, EPHB2 (p = 0.000542) and PDZK1IP1 (p = 0.0206) with RA-specific gene expression profiles and elevated expression of the ST6GALNAC1 (p = 0.0023) gene in UA. ROC and AUC risk score analysis suggested that MSA4A (AUC: 0.894, 0.644, 0.720), PDZK1IP1 (AUC: 0.785, 0.806, 0.977), and EPHB2 (AUC: 0.794, 0.723, 0.620) at 0, 6, and 12 months follow-up can accurately discriminate patients with RA from healthy controls and may have practical value for RA diagnosis. In patients with early inflammatory arthritis, ST6GALNAC1 is a potential biomarker for UA as compared with healthy controls whereas EPHB2, MS4A4A, and particularly PDZK1IP1 may discriminate RA patients. SIGLEC1 may also be a useful marker of disease activity in UA.
Detecting cardiac pathologies via machine learning on heart-rate variability time series and related markers
Authors: Agliari Elena , Barra Adriano , Barra Orazio Antonio , Fachechi Alberto , Franceschi Vento Lorenzo , Moretti Luciano
Published: June 2020

In this paper we develop statistical algorithms to infer possible cardiac pathologies, based on data collected from 24 h Holter recording over a sample of 2829 labelled patients; labels highlight whether a patient is suffering from cardiac pathologies. In the first part of the work we analyze statistically the heart-beat series associated to each patient and we work them out to get a coarse-grained description of heart variability in terms of 49 markers well established in the reference community. These markers are then used as inputs for a multi-layer feed-forward neural network that we train in order to make it able to classify patients. However, before training the network, preliminary operations are in order to check the effective number of markers (via principal component analysis) and to achieve data augmentation (because of the broadness of the input data). With such groundwork, we finally train the network and show that it can classify with high accuracy (at most ~85% successful identifications) patients that are healthy from those displaying atrial fibrillation or congestive heart failure. In the second part of the work, we still start from raw data and we get a classification of pathologies in terms of their related networks: patients are associated to nodes and links are drawn according to a similarity measure between the related heart-beat series. We study the emergent properties of these networks looking for features (e.g., degree, clustering, clique proliferation) able to robustly discriminate between networks built over healthy patients or over patients suffering from cardiac pathologies. We find overall very good agreement among the two paved routes.
Destabilization of light NREM sleep by thalamic PLCβ4 deletion impairs sleep-dependent memory consolidation
Authors: Hong Joohyeon , Ha Go Eun , Kwak Hankyul , Lee Yelin , Jeong Hyeonyeong , Suh Pann-Ghill , Cheong Eunji
Published: June 2020

Sleep abnormality often accompanies the impairment of cognitive function. Both rapid eye movement (REM) and non-REM (NREM) sleep have associated with improved memory performance. However, the role of composition in NREM sleep, consisting of light and deep NREM, for memory formation is not fully understood. We investigated how the dynamics of NREM sleep states influence memory consolidation. Thalamocortical (TC) neuron-specific phospholipase C β4 (PLCβ4) knockout (KO) increased the total duration of NREM sleep, consisting of destabilized light NREM and stabilized deep NREM. Surprisingly, the longer NREM sleep did not improve memory consolidation but rather impaired it in TC-specific PLCβ4 KO mice. Memory function was positively correlated with the stability of light NREM and spindle activity occurring in maintained light NREM period. Our study suggests that a single molecule, PLCβ4, in TC neurons is critical for tuning the NREM sleep states and thus affects sleep-dependent memory formation.
Characterisation of a subpopulation of CD133+ cancer stem cells from Chinese patients with oral squamous cell carcinoma
Authors: Ma Zhen , Zhang Chong , Liu Xiaotong , Fang Fang , Liu Shiqi , Liao Xianxiang , Tao Shicheng , Mai Huaming
Published: June 2020

Cancer stem cells (CSCs) play a critical role in cancer development and growth. The aim of this study was to identify and isolate CSCs from populations of primary oral squamous cell carcinoma (OSCC) cells, which were obtained from OSCC specimens and identified by cell morphology and immunohistochemical staining for keratin. CD133+ cells detected by flow cytometry comprised 0.41 ± 0.06% of primary OSCC cells and were isolated from primary OSCC cell populations using magnetic-activated cell sorting, revealing that 93.39% of high-purity CD133+ cells were in the G0/G1 phase of the cell cycle. Additionally, the growth rate of CD133+ cells was higher than that of CD133− cells, and in vivo tumourigenesis experiments showed that the tumourigenic ability of CD133+ cells was markedly stronger than that of CD133− cells. Moreover, CD133+ cells showed increased chemotherapeutic resistance to cisplatin and higher self-renewal ability according to sphere-formation assay, as well as higher mRNA levels of stemness-associated genes, including NANOG, SOX2, ALDH1A1, and OCT4. These results indicated that OSCC cells, which share certain characteristics of CSCs, harbour CD133+ cells potentially responsible for OSCC aggressiveness, suggesting CD133 as a potential prognostic marker and therapeutic target.
Proanthocyanidin-rich grape seed extract improves bone loss, bone healing, and implant osseointegration in ovariectomized animals
Authors: Tenkumo Taichi , Aobulikasimu Alkebaier , Asou Yoshinori , Shirato Midori , Shishido Shunichi , Kanno Taro , Niwano Yoshimi , Sasaki Keiichi , Nakamura Keisuke
Published: June 2020

The purpose of the present study was to confirm if proanthocyanidin-rich grape seed extract (GSE) had the ability to improve bone health such as bone loss, bone healing, and implant osseointegration (defined as the direct connection between bone tissue and an implant) in ovariectomized (OVX) animals. We demonstrated that daily oral administration of GSE prevented bone loss in the lumbar vertebrae and femur in OVX mice. In addition, osteoclastogenesis in the lumbar spine bone of OVX mice, as assessed by histological and histomorphometric analyses, was accelerated but GSE prevented this dynamization, suggesting that GSE could counteract OVX-induced accelerated osteoclastogenic activity. In rats, OVX clearly impaired the healing of defects created on the calvaria, and GSE overcame this OVX-impaired healing. In the same way, osseointegration of a tibial implant in rats was retarded by OVX, and GSE counteracted the OVX-induced poor osseointegration, likely promoting bone healing by preventing imbalanced bone turnover. These results suggest that orally administered GSE improved implant osseointegration by mitigating the impaired bone health induced by OVX as a model of estrogen deficiency.