bluuebunny/crossref_metadata_2025 huggingface dataset:
Gamma Frequency Sensory Stimulation in Probable Mild Alzheimer’s Dementia Patients: Results of a Preliminary Clinical Trial
Authors: Chan Diane , Suk Ho-Jun , Jackson Brennan , Milman Noah Pollak , Stark Danielle , Klerman Elizabeth B. , Kitchener Erin , Avalos Vanesa S. Fernandez , Banerjee Arit , Beach Sara D. , Blanchard Joel , Stearns Colton , Boes Aaron , Uitermarkt Brandt , Gander Phillip , Howard Matthew , Sternberg Eliezer J. , Nieto-Castanon Alfonso , Anteraper Sheeba , Whitfield-Gabrieli Susan , Brown Emery N. , Boyden Edward S. , Dickerson Bradford , Tsai Li-Huei
Published: March 2021

Non-invasive Gamma ENtrainment Using Sensory stimuli (GENUS) at 40Hz reduced Alzheimer’s disease (AD) pathology, prevented cerebral atrophy and improved performance during behavioral testing in mouse models of AD. We report data from a safety study (NCT04042922) and a randomized, placebo-controlled trial in participants with probable mild AD dementia after 3 months of one-hour daily 40Hz light and sound GENUS (NCT04055376) to assess safety, compliance, entrainment and possible effects on brain structure, function, sleep and cognitive function. GENUS was well-tolerated and compliance was high in both groups. Electroencephalography recordings show that our GENUS device safely and effectively induced 40Hz entrainment in cognitively normal subjects and participants with mild AD. After 3 months of daily stimulation, participants with mild AD in the 40Hz GENUS group showed less ventricular enlargement and stabilization of the hippocampal size compared to the control group. Functional connectivity increased in both the default mode network and the medial visual network after 3 months of stimulation. Circadian rhythmicity also improved with GENUS. Compared to controls, the active group performed better on the face-name association delayed recall test. These results suggest that 40Hz GENUS can be used safely at home daily and shows favorable outcomes on cognitive function, daily rhythms, and structural and functional MRI biomarkers of AD-related degeneration. These results support further evaluation of GENUS in larger and longer clinical trials to evaluate its potential as a disease modifying therapeutic for Alzheimer’s disease.
Metatranscriptomic comparison of endophytic and pathogenicFusarium–Arabidopsis interactions reveals plant transcriptional plasticity
Authors: Guo Li , Yu Houlin , Wang Bo , Vescio Kathryn , DeIulio Gregory A. , Yang He , Berg Andrew , Zhang Lili , Edel-Hermann Véronique , Steinberg Christian , Kistler H. Corby , Ma Li-Jun
Published: March 2021

Plants are continuously exposed to beneficial and pathogenic microbes, but how plants recognize and respond to friends versus foes remains poorly understood. Here, we compared the molecular response ofArabidopsis thalianaindependently challenged with aFusarium oxysporumendophyte Fo47 versus a pathogen Fo5176. These twoFusarium oxysporumstrains share a core genome of about 46 Mb, in addition to unique 1,229 and 5,415 accessory genes. Metatranscriptomic data reveal a shared pattern of expression for most plant genes (∼80%) in responding to both fungal inoculums at all time points from 12 to 96 h post inoculation (HPI). However, the distinct responding genes depict transcriptional plasticity, as the pathogenic interaction activates plant stress responses and suppresses plant growth/development related functions, while the endophytic interaction attenuates host immunity but activates plant nitrogen assimilation. The differences in reprogramming of the plant transcriptome are most obvious in 12 HPI, the earliest time point sampled and are linked to accessory genes in both fungal genomes. Collectively, our results indicate that theA. thalianaandF. oxysporuminteraction displays both transcriptome conservation and plasticity in the early stages of infection, providing insights into the fine-tuning of gene regulation underlying plant differential responses to fungal endophytes and pathogens.

One-sentence summary

Multiomics analysis reveals the regulatory plasticity of plants in response to beneficial and antagonistic microbes, resulting in distinct phenotypes and rewired transcriptional networks.
COMPARISON OF sPLA2-IIA PERFORMANCE WITH HIGH-SENSITIVE CRP, NEUTROPHIL PERCENTAGE, PCT AND LACTATE TO IDENTIFY BACTERIAL INFECTION: A PROSPECTIVE STUDY
Authors: Tan Toh Leong , Kang Christabel Wan-li , Ooi Kai Shen , Tan Swee Thian , Ahmad Nurul Saadah , Nasuruddin Dian Nasriana , Ithnin Azlin , Arifin Khaizurin Tajul , Lee Yook Heng , Hassan Nurul Izzaty , Gan Kok Beng , Neoh Hui-min
Published: March 2021

Early bacterial infection (BI) identification in resource-limiting Emergency Departments (ED) is challenging especially in low- and middle-income counties (LMIC). Misdiagnosis predisposes to antibiotic overuse and propagates antimicrobial resistance. This study evaluates new emerging biomarkers, secretory phospholipase A2 group IIA (sPLA2-IIA), and compares with other biomarkers on their performance characteristic of BI detection in Malaysia, an LMIC. A prospective cohort study was conducted involving 151 consecutive patients admitted to the ED. A single measurement was taken upon patient arrival in ED and was analysed for serum levels of sPLA2-IIA, high-sensitive C-reactive protein (CRP), procalcitonin (PCT), neutrophil percentage (N%), and lactate. All biomarkers’ performance was compared for the outcomes using area under the receiver operating characteristic curve (AUROC), sensitivity, and specificity. The performance of sPLA2-IIA (AUROC 0.93 [95% CI: 0.89-0.97]; Sn 80% [95% CI: 72-87] Sp 94% [95% CI: 81-89]) was the highest among all. It was comparable with high-sensitive CRP (AUROC 0.93 [95% CI: 0.88-0.97]; Sn 75% [95% CI: 66-83]; Sp 91 [95% CI: 77-98]) but had a higher Sn and Sp. The sPLA2-IIA was also found superior to N%, PCT, and lactate. This finding suggested sPLA2-IIA was recommended biomarkers for BI detection in LMIC.
Partial agonism improves the anti-hyperglycaemic efficacy of an oxyntomodulin-derived GLP-1R/GCGR co-agonist
Authors: Pickford Phil , Lucey Maria , Rujan Roxana-Maria , McGlone Emma Rose , Bitsi Stavroula , Ashford Fiona , Corrêa Ivan R , Hodson David J , Tomas Alejandra , Deganutti Giuseppe , Reynolds Christopher A , Owen Bryn M , Tan Tricia M , Minnion James , Jones Ben , Bloom Stephen R
Published: March 2021

Objective

Glucagon-like peptide-1 and glucagon receptor (GLP-1R/GCGR) co-agonism can maximise weight loss and improve glycaemic control in type 2 diabetes and obesity. In this study we investigated the cellular and metabolic effects of modulating the balance between G protein activation and β-arrestin-2 recruitment at GLP-1R and GCGR using oxyntomodulin (OXM)-derived co-agonists. This strategy has been previously shown to improve the duration of action of GLP-1R mono-agonists by reducing target desensitisation and downregulation.

Methods

Dipeptidyl dipeptidase-4 (DPP-4)-resistant OXM analogues were generated and assessed for a variety of cellular readouts. Molecular dynamic simulations were used to gain insights into the molecular interactions involved. In vivo studies were performed in mice to identify effects on glucose homeostasis and weight loss.

Results

Ligand-specific reductions in β-arrestin-2 recruitment led to reduced GLP-1R internalisation and prolonged glucose-lowering actionin vivo. The putative benefits of GCGR agonism were retained, with equivalent weight loss compared to the GLP-1R mono-agonist liraglutide in spite of a lesser degree of food intake suppression. The compounds tested showed only a minor degree of biased agonism between G protein and β-arrestin-2 recruitment at both receptors and were best classified as partial agonists for the two pathways measured.

Conclusions

Diminishing β-arrestin-2 recruitment may be an effective way to increase the therapeutic efficacy of GLP-1R/GCGR co-agonists. These benefits can be achieved by partial rather than biased agonism.
Metabolic dysfunction induced by high-fat diet modulates hematopoietic stem and myeloid progenitor cells in brown adipose tissue of mice
Authors: Mincham Kyle T , Panchal Kunjal , Hart Prue H , Lucas Robyn M , Feelisch Martin , Weller Richard B , Matthews Vance B , Strickland Deborah H , Gorman Shelley
Published: March 2021

Brown adipose tissue (BAT) may be an important metabolic regulator of whole-body glucose. While important roles have been ascribed to macrophages in regulating metabolic functions in BAT, little known is known of the roles of other immune cells subsets, particularly dendritic cells (DCs). Eating a high fat diet may compromise the development of hematopoietic stem and progenitor cells (HSPC) – which give rise to DCs – in bone marrow, with less known of its effects in BAT. We have previously demonstrated that ongoing exposure to low-dose ultraviolet radiation (UVR) significantly reduced the ‘whitening’ effect of eating a high-fat diet upon interscapular (i)BAT of mice. Here, we examined whether this observation may be linked to changes in the phenotype of HSPC and myeloid-derived immune cells in iBAT and bone marrow of mice using 12-colour flow cytometry. Many HSPC subsets declined in both iBAT and bone marrow with increasing metabolic dysfunction. Conversely, with rising adiposity and metabolic dysfunction, conventional (c)DCs increased in both of these tissues. When compared to low-fat diet, consumption of high-fat diet significantly reduced proportions of myeloid, common myeloid and megakaryocyte-erythrocyte progenitors in iBAT, and short-term hematopoietic stem cells in bone marrow. In mice fed a high-fat diet, exposure to low-dose UVR significantly reduced proportions of cDCs in iBAT, independently of nitric oxide release from irradiated skin (blocked using the scavenger, cPTIO), but did not significantly modify HSPC subsets in either tissue. Further studies are needed to determine whether changes in these cell populations contribute towards metabolic dysfunction.
Drug-driven reclassification of multiple tumour subtypes reveals intrinsic molecular concordance of therapy across histologically disparate cancers
Authors: Xu Yue , Zheng Jie , Cai Zhaoqing , Li Wang , Köhler Jens , Dai Yao , Cheng Xiaojie , Wu Tao , Zhang Fan , Wang Haiyun
Published: March 2021

Cancers that are histologically defined as the same type of cancer often need a distinct therapy based on underlying heterogeneity; likewise, histologically disparate cancers can require similar treatment approaches due to intrinsic similarities. A comprehensive analysis integrated with drug response data and genomic alterations, particularly to reveal therapeutic concordance mechanisms across histologically disparate tumour subtypes, has not yet been fully exploited.

In this study, we used pharmacogenomic profiling data provided from the Cancer Genome Project (CGP) in a systematic in silico investigation of the pharmacological subtypes of cancers and the intrinsic concordance of molecular mechanisms leading to similar therapeutic responses across histologically disparate tumour subtypes. We further developed a novel approach to redefine cell-to-cell similarity and drug-to-drug similarity from the therapeutic concordance, providing a new point of view to study cancer heterogeneity.

Our study identified that histologically different tumours, such as malignant melanoma and colorectal adenocarcinoma, could belong to the same pharmacological subtype regarding drug sensitivity to MEK inhibitors, which was determined by their genomic alterations, high occurrence of BRAF or KRAS mutations. Therapeutic concordance for chemotherapy drugs was identified across histologically disparate hematological tumors mainly due to the extraordinary activation of the cell cycle in blood cancers. A subcluster of SCLC had a more similar profile with hematological tumors, and was associated with the malignant phenotype, with a higher level of MYC expression. We developed a website to store and visualize the pharmacological subtypes of drugs, as well as their connected genomic and expression alterations.
Intrinsic cardiac adrenergic cells contribute to septic cardiomyopathy
Authors: Yang Duomeng , Dai Xiaomeng , Xing Yun , Tang Xiangxu , Yang Guang , Wang Penghua , Harrison Andrew G. , Li Hongmei , Lv Xiuxiu , Yu Xiaohui , Wang Huadong
Published: March 2021

Occurring independently of cardiac sympathetic nervous system, the intrinsic cardiac adrenergic (ICA) cells have been identified as an important regulator in both of developing and adult cardiac physiological and pathological processes. However, its role in septic cardiomyopathy remains unknown. Herein, we report that lipopolysaccharide (LPS) dose- and time-dependently increased norepinephrine (NE) release from ICA cells, which aggravates myocardial TNF-α production and dysfunction. Inhibition of NE synthesis in ICA cells alleviated LPS-elicited cardiac dysfunction as well as TNF-α production in Langendorff perfusing hearts. Mechanistically, ICA cell expressed Toll-like receptor 4 (TLR4), activated by LPS, to increase the expression of tyrosine hydroxylase, a key enzyme responsible for NE biosynthesis, via AP-1 binding to its promoter. Surprisingly, LPS-TLR4 signaling triggered no TNF-α production in ICA cells due to the elevatedNfkbiaandTnfaip6expression. In LPS-treated co-culture of ICA cells and cardiomyocytes, the raised NE from ICA cells activated cardiomyocyte β1-adrenergic receptor (β1-AR), driving Ca2+/calmodulin-dependent protein kinase II (CaMKII) to increase the activities of NF-κB and mitogen-activated protein kinase pathways, which were mimicked by dobutamine. Our findings reveal a cell type-specific TLR4 function triggering NE synthesis, but not TNF-α production in inflammatory pathogenesis, and identify ICA cell-derived NE as a paracrine signal in the cross talk among different cardiac cells to enhance myocardial injury during LPS challenge, suggesting that targeting ICA cell-derived NE may be a potential therapeutic strategy for septic cardiomyopathy.
A CTDNEP1-lipin 1-mTOR regulatory network restricts ER membrane biogenesis to enable chromosome motions necessary for mitotic fidelity
Authors: Merta Holly , Carrasquillo Rodríguez Jake W. , Anjur-Dietrich Maya I. , Granade Mitchell E. , Vitale Tevis , Harris Thurl E. , Needleman Daniel J. , Bahmanyar Shirin
Published: March 2021

The endoplasmic reticulum (ER) dramatically restructures in open mitosis to become excluded from the mitotic spindle; however, the significance of ER reorganization to mitotic progression is not known. Here, we demonstrate that limiting ER membrane biogenesis enables mitotic chromosome movements necessary for chromosome biorientation and prevention of micronuclei formation. Aberrantly expanded ER membranes increase the effective viscosity of the mitotic cytoplasm to physically restrict chromosome dynamics – slowed chromosome motions impede correction of mitotic errors induced by transient spindle disassembly, leading to severe micronucleation. We define the mechanistic link between regulation of ER membrane biogenesis and mitotic fidelity by demonstrating that a CTDNEP1-lipin 1-mTOR regulatory network limits ER lipid synthesis to prevent chromosome missegregation. Together, this work shows that ER membranes reorganize in mitosis to enable chromosome movements necessary for mitotic error correction and reveal dysregulated lipid metabolism as a potential source of aneuploidy in cancer cells.
Chromatin Network Retards Droplet Coalescence
Authors: Qi Yifeng , Zhang Bin
Published: March 2021

Nuclear bodies are membraneless condensates that may form via liquid-liquid phase separation. The viscoelastic chromatin network could impact their stability and may hold the key for understanding experimental observations that defy predictions of classical theories. However, quantitative studies on the role of the chromatin network in phase separation have remained challenging. Using a diploid human genome model parameterized with chromosome conformation capture (Hi-C) data, we studied the thermodynamics and kinetics of droplet formation inside the nucleus. Dynamical simulations predicted the formation of multiple droplets for protein particles that experience specific interactions with nucleolus-associated domains (NADs). Coarsening dynamics, surface tension, and coalescence kinetics of the simulated droplets are all in quantitative agreements with experimental measurements for nucleoli. Free energy calculations further supported that a two-droplet state, which is often observed for nucleoli seen in somatic cells, is metastable and separated from the single-droplet state with an entropic barrier. Our study suggests that protein-chromatin interactions facilitate the nucleation of droplets, but hinders their coarsening due to the correlated motion between droplets and the chromatin network: as droplets coalesce, the chromatin network becomes increasingly constrained. Therefore, protein-chromatin interactions arrest phase separation in multi-droplet states and may drive the variation of nuclear body numbers across cell types.
Multi-dimensional and spatiotemporal correlative imaging at the plasma membrane of live cells to determine the continuum nano-to-micro scale lipid adaptation and collective motion
Authors: Bernabé-Rubio Miguel , Bosch-Fortea Minerva , Alonso Miguel A. , de la Serna Jorge Bernardino
Published: March 2021

The primary cilium is a specialized plasma membrane protrusion with important receptors for signalling pathways. In polarized epithelial cells, the primary cilium assembles after the midbody remnant (MBR) encounters the centrosome at the apical surface. The membrane surrounding the MBR, namely remnant associated membrane patch (RAMP) once situated next to the centrosome, releases some of its lipid components to form a centrosome-associated membrane patch (CAMP) from which the ciliary membrane stems. The RAMP undergoes a spatiotemporal membrane refinement during the formation of the CAMP, which becomes highly enriched in condensed membranes with low lateral mobility. To better understand this process, we have developed a correlative imaging approach that yields quantitative information about the lipid lateral packing, its mobility and collective assembly at the plasma membrane at different spatial scales over time. Our work paves the way towards a quantitative understanding of lipid collective assembly at the plasma membrane spatiotemporally as a functional determinant in cell biology and its direct correlation with the membrane physicochemical state. These findings allowed us to gain a deeper insight into the mechanisms behind the biogenesis of the ciliary membrane of polarized epithelial cells.
A single-cell atlas of breast cancer cell lines to study tumour heterogeneity and drug response
Authors: Gambardella G , Viscido G , Tumaini B , Isacchi A , Bosotti R , di Bernardo D
Published: March 2021

Breast cancer patient stratification is mainly driven by tumour receptor status and histological grading and subtyping, with about twenty percent of patients for which absence of any actionable biomarkers results in no clear therapeutic intervention. Cancer cells within the same tumour have heterogeneous phenotypes and exhibit dynamic plasticity. However, how to evaluate such heterogeneity and its impact on outcome and drug response is still unclear. Here, we transcriptionally profiled 35,276 individual cells from 32 breast cancer cell lines covering all main breast cancer subtypes to yield a breast cancer cell line atlas. We found high degree of heterogeneity in the expression of clinically relevant biomarkers across individual cells within the same cell line; such heterogeneity is non-genetic and dynamic. We computationally mapped single cell transcriptional profiles of patients’ tumour biopsies to the atlas to determine their composition in terms of cell lines. Each tumour was found to be heterogenous and composed of multiple cell lines mostly, but not exclusively, of the same subtype. We then trained an algorithm on the atlas to determine cell line composition from bulk gene expression profiles of tumour biopsies, thus providing a novel approach to patient stratification. Finally, we linked results from large-scale in vitro drug screening1,2to the single cell data to computationally predict responses to more than 450 anticancer agents starting from single-cell transcriptional profiles. We thus found that transcriptional heterogeneity enables cells with differential drug sensitivity to co-exist in the same population. Our work provides a unique resource and a novel framework to determine tumour heterogeneity and drug response in breast cancer patients.
The glycocalyx affects the mechanotransductive perception of the topographical microenvironment
Authors: Chighizola Matteo , Dini Tania , Marcotti Stefania , D’Urso Mirko , Piazzoni Claudio , Borghi Francesca , Previdi Anita , Ceriani Laura , Folliero Claudia , Stramer Brian , Lenardi Cristina , Milani Paolo , Podestà Alessandro , Schulte Carsten
Published: March 2021

The cell/microenvironment interface is the starting point of integrin-mediated mechanotransduction, but many details of mechanotransductive signal integration remain elusive due to the complexity of the involved (extra)cellular structures, such as the glycocalyx.

We used nano-bio-interfaces reproducing the complex nanotopographical features of the extracellular matrix to analyse the glycocalyx impact on PC12 cell mechanosensing at the nanoscale (e.g., by force spectroscopy with functionalised probes). Our data demonstrates that the glycocalyx configuration affects spatio-temporal nanotopography-sensitive mechanotransductive events at the cell/microenvironment interface. Opposing effects of glycocalyx removal were observed, when comparing flat and specific nanotopographical conditions. The excessive retrograde actin flow speed and force loading are strongly reduced on certain nanotopographies upon removal of the native glycocalyx, while on the flat substrate we observe the opposite trend.

Our results highlight the importance of the glycocalyx configuration in a molecular clutch force loading-dependent cellular mechanism for mechanosensing of microenvironmental nanotopographical features.
Neutrophils require SKAP2 for reactive oxygen species production following C-type lectin andCandidastimulation
Authors: Nguyen Giang T. , Xu Shuying , Bunnell Stephen C. , Mansour Michael K. , Sykes David B. , Mecsas Joan
Published: March 2021

Signaling cascades that convert the recognition of pathogens to efficient inflammatory responses by immune cells, specifically neutrophils, are critical for host survival. SKAP2, an adaptor protein, is required for reactive oxygen species (ROS) generation following stimulation by integrins, formyl peptide receptors and gram-negative bacteriaKlebsiella pneumoniaeandYersinia pseudotuberculosis in vitro(Nguyen et al., 2020, Shaban et al., 2020, Boras et al., 2017). SKAP2 is also required for the host defense againstK. pneumoniaeandΔyopH Y. pseudotuberculosisinfectionin vivoin mouse models (Shaban et al., 2020, Nguyen et al., 2020). Another class of pattern recognition receptors (PRR) is the C-type lectin receptors (CLR), such as Dectin-1, Dectin-2 and Mincle, that are critical to trigger innate immune responses. Using neutrophils from murine HoxB8-immortalized progenitors, we show that SKAP2 is crucial for maximal ROS response to purified CLR agonists and to the fungal pathogensCandida glabrataandC. albicans, as well as for robust killing ofC. glabrata.Skap2-/-murine neutrophils failed to generate ROS and exhibited reduced cellular adhesion in response to trehalose-6,6’-dibehenate (TDB), furfurman, and curdlan, Mincle, Dectin-2, and Dectin-1 agonists, respectively. TDB, furfurman, and curdlan stimulation also led to SKAP2-independent integrin conformational changes, showing that inside-out signaling by these CLRs to integrin occurs in the absence of SKAP2. Pyk2 phosphorylation was significantly reduced after infection withC. glabratainSkap2-/-neutrophils, while Syk phosphorylation was unaffected by the loss of SKAP2. These data strengthen the importance of SKAP2 in the activation of neutrophil ROS production by PRRs to include CLRs and extend the role of SKAP2 in host defense beyond antibacterial immunity to includeCandidaspecies.
A Pharmacophore Model for SARS-CoV-2 3CLpro Small Molecule Inhibitors and in Vitro Experimental Validation of Computationally Screened Inhibitors
Authors: Glaab Enrico , Manoharan Ganesh Babu , Abankwa Daniel
Published: March 2021

Among the biomedical efforts in response to the current coronavirus (COVID-19) pandemic, pharmacological strategies to reduce viral load in patients with severe forms of the disease are being studied intensively. One of the main drug target proteins proposed so far is the SARS-CoV-2 viral protease 3CLpro (also called Mpro), an essential component for viral replication. Ongoing ligand- and receptor-based computational screening efforts would be facilitated by an improved understanding of the electrostatic, hydrophobic and steric features that characterize small molecule inhibitors binding stably to 3CLpro, as well as by an extended collection of known binders.

Here, we present combined virtual screening, molecular dynamics simulation, machine learning andin vitroexperimental validation analyses which have led to the identification of small molecule inhibitors of 3CLpro with micromolar activity, and to a pharmacophore model that describes functional chemical groups associated with the molecular recognition of ligands by the 3CLpro binding pocket. Experimentally validated inhibitors using a ligand activity assay include natural compounds with available prior knowledge on safety and bioavailability properties, such as the natural compound rottlerin (IC50= 37 µM), and synthetic compounds previously not characterized (e.g. compound CID 46897844, IC50= 31 µM). In combination with the developed pharmacophore model, these and other confirmed 3CLpro inhibitors may provide a basis for further similarity-based screening in independent compound databases and structural design optimization efforts, to identify 3CLpro ligands with improved potency and selectivity.

Overall, this study suggests that the integration of virtual screening, molecular dynamics simulations and machine learning can facilitate 3CLpro-targeted small molecule screening investigations. Different receptor-, ligand- and machine learning-based screening strategies provided complementary information, helping to increase the number and diversity of identified active compounds. Finally, the resulting pharmacophore model and experimentally validated small molecule inhibitors for 3CLpro provide resources to support follow-up computational screening efforts for this drug target.
A heterohexameric protein consisting of six linked single-domain antibodies is highly protective for BoNT/A, BoNT/B and BoNT/E exposures
Authors: Mukherjee Jean , Tremblay Jacqueline M. , Debatis Michelle , Foss Alexa , Awata Junya , Shoemaker Charles B.
Published: March 2021

Botulinum neurotoxin (BoNT) serotypes A, B and E cause the vast majority of human botulism cases and pose the greatest bioterrorism threats. We previously identified multiple camelid single-domain antibodies (VHHs) that each neutralize BoNT/A, BoNT/B or BoNT/E. We also demonstrated that heterodimers of linked toxin-neutralizing VHHs are much more potent than VHH monomer pools in preventing BoNT intoxication. In this study, we expressed two different heterohexamer proteins (VNA1-ABE and VNA2-ABE) of ~100 kDa secreted from mammalian host cells, each containing the same six linked anti-BoNT VHH components ordered in two different combinations. Each heterohexamer contained two VHHs that neutralize BoNT/A, BoNT/B or BoNT/E. Both heterohexameric antitoxins displayed similar strong binding properties for the three targeted BoNT serotypes by ELISA. One ug of each heterohexameric antitoxin fully protected groups of mice co-administered with 100 LD50of BoNT/A, BoNT/B or BoNT/E, or a pool containing 100 LD50of each of the three toxins. The results demonstrate that long chains of at least six different linked VHHs can be expressed such that all component VHHs in the multimer retain their target binding activities. These findings make more feasible the development of a BoNT antitoxin product consisting of a small pool of proteins that, in combination, neutralize all known BoNT serotypes and subtypes.

Key Contribution

Heteromultimeric proteins consisting of six linked, VHH antibodies, and including VHHs that neutralize BoNT/A, BoNT/B and BoNT/E, retain high potency to protect mice challenged with high doses of all three of these BoNT serotypes.
Desmosomal dualism: the core is stable while plakophilin is dynamic
Authors: Fülle Judith B , Huppert Henri , Liebl David , Liu Jaron , Almeida Rogerio Alves de , Yanes Bian , Wright Graham D , Lane E Birgitte , Garrod David R , Ballestrem Christoph
Published: March 2021

Desmosomes, strong cell-cell junctions of epithelia and cardiac muscle, link intermediate filaments to cell membranes and mechanically integrate cells across tissues, dissipating mechanical stress. They comprise 5 major protein classes - desmocollins and desmogleins (the desmosomal cadherins), plakoglobin, plakophilins and desmoplakin - whose individual contribution to the structure and turnover of desmosomes is poorly understood. Using live-cell imaging together with FRAP and FLAP we show that desmosomes consist of two contrasting protein fractions or modules: a very stable desmosomal core of desmosomal cadherins and plakoglobin, and a highly mobile plakophilin. As desmosomes mature from calcium-dependence to calciumindependent hyper-adhesion, core stability increases, but Pkp2a remains highly mobile. Desmoplakin is initially mobile but stabilises with hyper-adhesion. We show that desmosome down-regulation during growth factor-induced cell scattering proceeds by internalisation of whole desmosomes, which still retain a stable core and highly mobile Pkp2a. This molecular mobility of Pkp2a suggests a transient and probably regulatory role for Pkp2a in the desmosome.
Enabling single-molecule localization microscopy in turbid food emulsions
Authors: Jabermoradi Abbas , Yang Suyeon , Gobes Martijn , van Duynhoven John P.M. , Hohlbein Johannes
Published: March 2021

Turbidity poses a major challenge for the microscopic characterization of many food systems. In these systems, local mismatches in refractive indices can cause reflection, absorption and scattering of incoming as well as outgoing light leading to significant image deterioration along sample depth. To mitigate the issue of turbidity and to increase the achievable optical resolution, we combined adaptive optics (AO) with single-molecule localization microscopy (SMLM). Building on our previously published open hardware microscopy framework, the miCube, we first added a deformable mirror to the detection path. This element enables both the compensation of aberrations directly from single-molecule data and, by further modulating the emission wavefront, the introduction of various point spread functions (PSFs) to enable SMLM in three dimensions. We further added a top hat beam shaper to the excitation path to obtain an even illumination profile across the field of view (FOV). As a model system for a non-transparent food colloid in which imaging in depth is challenging, we designed an oil-in-water emulsion in which phosvitin, a ferric ion binding protein present in from egg yolk, resides at the oil water interface. We targeted phosvitin with fluorescently labelled primary antibodies and used PSF engineering to obtain 2D and 3D images of phosvitin covered oil droplets with sub 100 nm resolution. Droplets with radii as low as 200 nm can be discerned, which is beyond the range of conventional confocal light microscopy. Our data indicated that in the model emulsion phosvitin is homogeneously distributed at the oil-water interface. With the possibility to obtain super-resolved images in depth of nontransparent colloids, our work paves the way for localizing biomacromolecules at colloidal interfaces in heterogeneous food emulsions.
Crystal structures of anthranilate phosphoribosyltransferase from Saccharomyces cerevisiae
Authors: Wu Xiaofei , Zhang Mengying , Kuang Zhiling , Yue Jian , Xue Lu , Zhu Min , Zhu Zhongliang , Khan Muhammad Hidayatullah , Niu Liwen
Published: March 2021

Anthranilate phosphoribosyltransferase (AnPRT) catalyzes the transfer of the phosphoribosyl group of 5′-phosphoribosyl-1′-pyrophosphate (PRPP) to anthranilate to form phosphoribosyl-anthranilate. Crystal structures of AnPRTs from bacteria and archaea have previously been determined; however, the structure of Saccharomyces cerevisiae AnPRT (ScAnPRT) still remains unsolved. Here, crystal structures of ScAnPRT in the apo form as well as in complex with its substrate PRPP and the substrate analogue 4-fluoroanthranilate (4FA) are presented. These structures demonstrate that ScAnPRT exhibits the conserved structural fold of type III phosphoribosyltransferase enzymes and shares the similar mode of substrate binding found across the AnPRT protein family. In addition, crystal structures of ScAnPRT mutants (ScAnPRTSer121Ala and ScAnPRTGly141Asn) were also determined. These structures suggested that the conserved residue Ser121 is critical for binding PRPP, while Gly141 is dispensable for binding 4FA. In summary, these structures improved the preliminary understanding of the substrate-binding mode of ScAnPRT and laid foundations for future research.
Crystal structure of human CRM1, covalently modified by 2-mercaptoethanol on Cys528, in complex with RanGTP
Authors: Shaikhqasem Alaa , Schmitt Kerstin , Valerius Oliver , Ficner Ralf
Published: March 2021

CRM1 is a nuclear export receptor that has been intensively targeted over the last decade for the development of antitumor and antiviral drugs. Structural analysis of several inhibitor compounds bound to CRM1 revealed that their mechanism of action relies on the covalent modification of a critical cysteine residue (Cys528 in the human receptor) located in the nuclear export signal-binding cleft. This study presents the crystal structure of human CRM1, covalently modified by 2-mercaptoethanol on Cys528, in complex with RanGTP at 2.58 Å resolution. The results demonstrate that buffer components can interfere with the characterization of cysteine-dependent inhibitor compounds.
Re-examining the impact of financial intermediation on economic growth: evidence from Turkey
Authors: Yakubu Ibrahim Nandom , Abokor Aziza Hashi , Gedik Balay Iklim
Published: March 2021

Purpose

This study seeks to investigate the impact of financial intermediation on economic growth in Turkey using annual data spanning 1970–2017.

Design/methodology/approach

Based on the results of the augmented Dickey–Fuller and Phillips–Perron unit root tests for stationarity, the authors employ the Autoregressive Distributed Lag (ARDL) bounds testing to cointegration to establish the long-run impact of financial intermediation alongside other control factors on economic growth. The study also examines the short-run relationship between financial intermediation and economic growth by estimating the Error Correction Model (ECM).

Findings

The authors’ findings indicate that financial intermediation significantly influences economic growth in both short and long run. However, the effect is positive only in the short run, lending support to the supply-leading hypothesis. Regarding the control variables, the authors observe that while financial openness shows a positive significant impact on economic growth in the long run, gross fixed capital formation matters only in the short run. The results further infer that regardless of the time period, inflation impedes economic growth.

Originality/value

In the empirical analysis of the relationship between financial intermediation and economic growth, financial intermediation is always measured using a single variable. The authors argue that such studies could produce bias and misleading results given that a single proxy does not adequately reflect financial intermediation activities. Likewise, such findings may delude policy implementation. To provide a more vivid and robust analysis, the authors employ the Principal Component Analysis (PCA) to construct a composite index for financial intermediation based on three broad measures. The researchers’ are unaware of any study on the financial intermediation–economic growth nexus using a composite index of financial intermediation. Thus, this paper fills this lacuna in the literature.
Physico-chemical and mechanical properties of novel electrospun polyurethane composite with enhanced blood compatibility
Authors: Mani Mohan Prasath , Jaganathan Saravana Kumar
Published: March 2021

Purpose

This study aims to fabricate an electrospun scaffold by combining radish (Ra) and cerium oxide (CeO2) into a polyurethane (PU) matrix through electrospinning and investigate its feasibility for cardiac applications.

Design/methodology/approach

Physicochemical properties were analysed through various characterization techniques such as scanning electron microscopy (SEM), Fourier transforms infrared transforms analysis (FTIR), contact angle measurements, thermal analysis, atomic force microscopy (AFM) and mechanical testing. Further, blood compatibility assessments were carried out through activated partial thromboplastin time (APTT) and prothrombin time (PT) and hemolysis assay to evaluate the anticoagulant nature.

Findings

PU/Ra and PU/Ra/CeO2 exhibited a smaller fibre diameter than PU. Ra and CeO2 were intercalated in the polyurethane matrix which was evidenced in the infrared analysis by hydrogen bond formation. PU/Ra composite exhibited hydrophilic nature whereas PU/Ra/CeO2 composite turned hydrophobic. Surface measurements depicted the lowered surface roughness for the PU/Ra and PU/Ra/CeO2 compared to the pristine PU. PU/Ra and PU/Ra/CeO2 displayed enhanced degradation rates and improved mechanical strength than the pristine PU. The blood compatibility assay showed that the PU/Ra and PU/Ra/CeO2 had delayed blood coagulation times and rendered less toxicity against red blood cells (RBC’s) than PU.

Originality/value

This is the first report on the use of radish/cerium oxide in cardiac applications. The developed composite (PU/Ra and PU/Ra/CeO2) with enhanced mechanical and anticoagulant nature will serve as an indisputable candidate for cardiac tissue regeneration.
Winsome Joy: from educator to entrepreneur
Authors: MacNeil Heatherjean , Wiehe Lopes Amanda , Ozkazanc-Pan Banu , Douglass Anne
Published: March 2021

Research methodology

The information presented in this case was gathered through interviews and observations carried out during the time Ms Joy attended the Initiative for A Competitive Inner City business support program in 2017. In addition, focus groups that took place after the program provided important information and insights into her decision-making process and business goals. Additional interviews were conducted in 2018 and 2019 after the business program ended to gain in-depth knowledge of Ms Joy’s entrepreneurial journey.

Case overview/synopsis

This case details the experiences of Winsome Joy in recognizing market opportunities in the child care industry and then expanding into the educational materials industry. The case focuses on challenges related to founding and sustaining her business and the ways in which Ms Joy engaged in “opportunity recognition” and “effectuation” to become a successful entrepreneur. The case points out the challenges of the child care and early education field in terms of professional training, hiring practices and retaining qualified staff. It provides an aspirational role model who has overcome these challenges by finding and recognizing new market opportunities.

Complexity academic level

This case is relevant for undergraduate and graduate courses in entrepreneurship.
Body size and environment influence both intraspecific and interspecific variation in daily torpor use across hummingbirds
Authors: Spence Austin R. , Tingley Morgan W.
Published: March 2021

Torpor, or a regulated drop in body temperature and metabolic rate, allows animals to inhabit energetically costly environments, but among torpor‐using species, we have a poor understanding of how plasticity in torpor use relates to the experienced environment.

To better understand the ecology of daily torpor, we completed the largest study to date on the intraspecific variation of daily torpor use in hummingbirds by exposing 149 individuals of two hummingbird species to ambient or experimentally cooled temperatures in a field setting.

The smaller species, a latitudinal migrant, used daily torpor frequently under ambient conditions. The larger species, an elevational migrant, also used daily torpor regularly, but further increased the frequency of daily torpor use when experiencing colder temperatures and prior to migration—indicating a facultative adaptation.

To place our results within a broader phylogenetic context, we combined our experimental results with a meta‐analysis, including 31 species and all major hummingbird clades, and found a broad taxonomic pattern in which smaller hummingbirds are more likely to use daily torpor than their larger counterparts. Smaller hummingbirds may be physiologically constrained, requiring nearly obligate daily torpor use, while larger hummingbirds are physiologically more flexible and can facultatively respond to changing environmental conditions.

Our results reveal how physiological traits, such as the frequency and depth of daily torpor, can provide a mechanism to understand how hummingbird species have established and persisted across broad environmental gradients.

A free Plain Language Summary can be found within the Supporting Information of this article.
Non‐small‐cell lung cancer with ERBB2 mutation in non‐tyrosine kinase domain benefits from pyrotinib: A case report
Authors: Ni Jun , Si Xiao‐yan , Zhang Li
Published: March 2021

Tyrosine kinase domain (TKD) mutation and particularly exon 20 insertion mutations of erb‐b2 receptor tyrosine kinase 2 (ERBB2/HER2) have been extensively reported in non‐small cell lung cancer (NSCLC). Nevertheless, the clinical significance of non‐TKD mutations remains unknown. To date, no clinical trials have revealed that tyrosine kinase inhibitors are effective in NSCLC patients with non‐TKD ERBB2 mutations. Here we report a patient with advanced lung adenocarcinoma harboring non‐TKD mutation of ERBB2, S335C, without other actionable alterations benefited from pyrotinib. After first‐line treatment of pyrotinib monotherapy, a pan‐HER inhibitor, the patient achieved a durable partial response with good tolerance. This case powerfully illustrates that pyrotinib might be a promising first‐line treatment strategy for NSCLC patients with non‐TKD mutation of ERBB2.
COVID‐19 pandemic and allergen immunotherapy—an EAACI survey
Authors: Pfaar Oliver , Agache Ioana , Bonini Matteo , Brough Helen Annaruth , Chivato Tomás , Del Giacco Stefano R. , Gawlik Radoslaw , Gelincik Aslı , Hoffmann‐Sommergruber Karin , Jutel Marek , Klimek Ludger , Knol Edward F. , Lauerma Antti , Ollert Markus , O’Mahony Liam , Mortz Charlotte G. , Palomares Oscar , Riggioni Carmen , Schwarze Jürgen , Skypala Isabel , Torres María José , Untersmayr Eva , Walusiak‐Skorupa Jolanta , Chaker Adam , Giovannini Mattia , Heffler Enrico , Jensen‐Jarolim Erika , Quecchia Cristina , Sandoval‐Ruballos Mónica , Sahiner Umit , Tomić Spirić Vesna , Alvaro‐Lozano Montserrat
Published: March 2021

Background

As in many fields of medical care, the coronavirus disease 2019 (COVID‐19) resulted in an increased uncertainty regarding the safety of allergen immunotherapy (AIT). Therefore, the European Academy of Allergy and Clinical Immunology (EAACI) aimed to analyze the situation in different countries and to systematically collect all information available regarding tolerability and possible amendments in daily practice of sublingual AIT (SLIT), subcutaneous AIT (SCIT) for inhalant allergies and venom AIT.

Methods

Under the framework of the EAACI, a panel of experts in the field of AIT coordinated by the Immunotherapy Interest Group set‐up a web‐based retrospective survey (SurveyMonkey®) including 27 standardized questions on practical and safety aspects on AIT in worldwide clinical routine.

Results

417 respondents providing AIT to their patients in daily routine answered the survey. For patients (without any current symptoms to suspect COVID‐19), 60% of the respondents informed of not having initiated SCIT (40% venom AIT, 35% SLIT) whereas for the maintenance phase of AIT, SCIT was performed by 75% of the respondents (74% venom AIT, 89% SLIT). No tolerability concern arises from this preliminary analysis. 16 physicians reported having performed AIT despite (early) symptoms of COVID‐19 and/or a positive test result for severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2).

Conclusions

This first international retrospective survey in atopic diseases investigated practical aspects and tolerability of AIT during the COVID‐19 pandemic and gave no concerns regarding reduced tolerability under real‐life circumstances. However, the data indicate an undertreatment of AIT, which may be temporary, but could have a long‐lasting negative impact on the clinical care of allergic patients.
Carbohydrate epitopes currently recognized as targets for IgE antibodies
Authors: Platts‐Mills Thomas A. , Hilger Christiane , Jappe Uta , van Hage Marianne , Gadermaier Gabriele , Spillner Edzard , Lidholm Jonas , Keshavarz Behnam , Aalberse Rob C. , van Ree Ronald , Goodman Richard E. , Pomés Anna
Published: March 2021

Until recently, glycan epitopes have not been documented by the WHO/IUIS Allergen Nomenclature Sub‐Committee. This was in part due to scarce or incomplete information on these oligosaccharides, but also due to the widely held opinion that IgE to these epitopes had little or no relevance to allergic symptoms. Most IgE‐binding glycans recognized up to 2008 were considered to be “classical” cross‐reactive carbohydrate determinants (CCD) that occur in insects, some helminths and throughout the plant kingdom. Since 2008, the prevailing opinion on lack of clinical relevance of IgE‐binding glycans has been subject to a reevaluation. This was because IgE specific for the mammalian disaccharide galactose‐alpha‐1,3‐galactose (alpha‐gal) was identified as a cause of delayed anaphylaxis to mammalian meat in the United States, an observation that has been confirmed by allergists in many parts of the world. Several experimental studies have shown that oligosaccharides with one or more terminal alpha‐gal epitopes can be attached as a hapten to many different mammalian proteins or lipids. The classical CCDs also behave like haptens since they can be expressed on proteins from multiple species. This is the explanation for extensive in vitro cross‐reactivity related to CCDs. Because of these developments, the Allergen Nomenclature Sub‐Committee recently decided to include glycans as potentially allergenic epitopes in an adjunct section of its website (www.allergen.org). In this article, the features of the main glycan groups known to be involved in IgE recognition are revisited, and their characteristic structural, functional, and clinical features are discussed.
Bridging Materiality and Subjectivity: Expanding the Commons in Cooperation Birmingham
Authors: Ruiz Cayuela Sergio
Published: March 2021

Expansion is a matter of survival for emancipatory commons, permanently threatened by enclosure and cooptation. In this paper, I draw from my experience within Cooperation Birmingham to propose a theory (and practice) for expanding the commons that bridges two seemingly conflicting approaches. On the one hand, the concepts of “boundary commoning” and “commons ecologies” proposed by Massimo De Angelis, concerned with social reproduction and material autonomy. And on the other, “expanding commoning” as developed by Stavros Stavrides, which focuses on radical openness and the production of commoning subjectivities. I demonstrate how in organising a mutual aid response to the Covid‐19 pandemic, Cooperation Birmingham is using an expansion strategy that productively articulates both approaches. Boundary commoning and expanding commoning reinforce each other in the construction of a commons ecology that aims at posing a material alternative to capitalist social organisation while remaining always in‐the‐making and open to new commoners.