bluuebunny/crossref_metadata_2025_split huggingface dataset:
A luciferase prosubstrate and a red bioluminescent calcium indicator to image neuronal activity
Authors: Tian Xiaodong , Zhang Yiyu , Li Xinyu , Xiong Ying , Wu Tianchen , Ai Hui-wang 
Published: December 2021

AbstractGenetically encoded fluorescent indicators have been broadly used to monitor neuronal activity in live animals, but invasive surgical procedures are required. This study presents a functional bioluminescence imaging (fBLI) method for recording the activity of neuronal ensembles in the brain in awake mice. We developed a luciferase prosubstrate activatable in vivo by nonspecific esterase to enhance the brain delivery of the luciferin. We further engineered a bright, bioluminescent indicator with robust responsiveness to calcium ions (Ca2+) and appreciable emission above 600 nm. Integration of these advantageous components enabled the imaging of Ca2+ dynamics in awake mice minimally invasively with excellent signal- to-background and subsecond temporal resolution. This study thus establishes a new paradigm for studying brain functions in health and disease.
Endurance exercise ameliorates phenotypes in Drosophila models of Spinocerebellar Ataxias
Authors: Sujkowski Alyson , Richardson Kristin , Prifti Matthew V. , Wessells R. J. , Todi Sokol V. 
Published: December 2021

AbstractEndurance exercise is a potent intervention with widespread benefits proven to reduce disease incidence and impact across species. While endurance exercise supports neural plasticity, enhanced memory, and reduced neurodegeneration, less is known about the effect of chronic exercise on the progression of movement disorders such as ataxias. Here, we focused on three different types of ataxias, Spinocerebellar Ataxias Type (SCAs) 2, 3, and 6, belonging to the polyglutamine (polyQ) family of neurodegenerative disorders. In Drosophila models of these SCAs, flies progressively lose motor function. Here, we observe marked protection of speed and endurance in exercised SCA2 flies and modest protection in exercised SCA6 models, while no benefit is observed in SCA3 flies. Causative protein levels are reduced in SCA2 flies after chronic exercise, but not in SCA3 models, linking protein levels to exercise-based benefits. Additional investigations indicate that the exercise-inducible protein, Sestrin (Sesn) suppresses mobility decline and improves early death in SCA2 flies, even without exercise, coincident with disease protein level reduction and increased autophagic flux. These improvements depend on previously established functions of Sesn that reduce oxidative damage and modulate mTOR activity. Our study suggests differential responses of polyQ SCAs to exercise, highlighting the potential for more extensive application of exercise-based therapies in the prevention of polyQ neurodegeneration. Defining the mechanisms by which endurance exercise suppresses polyQ SCAs will open the door for more effective treatment for these diseases.
Oscillatory movement of a dynein-microtubule complex crosslinked with DNA origami
Authors: Abdellatef Shimaa A. , Tadakuma Hisashi , Yan Kangmin , Fujiwara Takashi , Fukumoto Kodai , Kondo Yuichi , Takazaki Hiroko , Boudria Rofia , Yasunaga Takuo , Higuchi Hideo , Hirose Keiko 
Published: December 2021

AbstractDuring repetitive bending of cilia and flagella, axonemal dynein molecules move in an oscillatory manner along a microtubule (MT), but how the minus-end-directed motor dynein can oscillate back and forth is unknown. There are various factors that may regulate the dynein activities, e.g., the nexin-dynein regulatory complex, radial spokes, and central apparatus. In order to understand the basic mechanism of the oscillatory movement, we constructed a simple model system composed of MTs, outer-arm dyneins, and DNA origami that crosslinks the MTs. Electron microscopy (EM) showed patches of dynein molecules crossbridging two MTs in two opposite orientations; the oppositely oriented dyneins are expected to produce opposing forces. The optical trapping experiments showed that the dynein-MT-DNA-origami complex actually oscillate back and forth after photolysis of caged ATP. Intriguingly, the complex, when held at one end, showed repetitive bending motions. The results show that a simple system composed of ensembles of oppositely oriented dyneins, MTs, and inter-MT crosslinkers, without the additional regulatory structures, has an intrinsic ability to cause oscillation and repetitive bending motions.
Visualizing cell death in live retina: Using calpain activity detection as a biomarker for retinal degeneration
Authors: Belhadj Soumaya , Hermann Nina , Christensen Gustav , Strasser Torsten , Paquet-Durand François 
Published: December 2021

AbstractCalpains are a family of calcium-activated proteases involved in numerous disorders. Notably, previous studies have shown that calpain activity was substantially increased in various models for inherited retinal degeneration (RD). In the present study, we tested the capacity of the t-BOC-Leu-Met-CMAC calpain-specific substrate to detect calpain activity in living retina, in organotypic retinal explant cultures derived from wild-type mice, as well as from rd1 and RhoP23H/+ RD-mutant mice. Test conditions were refined until the calpain substrate readily detected large numbers of cells in the photoreceptor layer of RD retina but not in wild-type retina. At the same time, the calpain substrate was not obviously toxic to photoreceptor cells. Comparison of calpain activity with an immunostaining for activated calpain-2 furthermore suggested that individual calpain isoforms may be active in distinct temporal stages of photoreceptor cell death. Notably, calpain-2 activity may be a relatively short-lived event, occurring only towards the end of the cell death process. Finally, our results support the development of calpain activity detection as a novel in vivo biomarker for RD, suitable for combination with non-invasive imaging techniques.
Azotobacter vinelandii scaffold protein NifU transfers iron to NifQ as part of the iron-molybdenum cofactor biosynthesis pathway for nitrogenase
Authors: Barahona Emma , Jiang Xi , Jiménez-Vicente Emilio , Rubio Luis M. , González-Guerrero Manuel 
Published: December 2021

ABSTRACTAzotobacter vinelandii molybdenum-dependent nitrogenase obtains molybdenum from NifQ, a monomeric iron-sulfur molybdoprotein. This protein requires of a preexisting [Fe-S] cluster to form a [MoFe3S4] group to serve as specific donor during nitrogenase cofactor biosynthesis. Here, we show biochemical evidence for NifU being the donor of the [Fe-S] cluster. Protein-protein interaction studies using apo-NifQ and as-isolated NifU demonstrated the interaction between both proteins which is only effective when NifQ is unoccupied by its [Fe-S] cluster. The apo-NifQ iron content increased after the incubation with as-isolated NifU, reaching similar levels to holo-NifQ after the interaction between apo-NifQ and NifU with reconstituted transient [Fe4-S4] groups. These results also indicate the necessity of co-expressing NifU together with NifQ in the pathway to provide molybdenum for the biosynthesis of nitrogenase in engineered nitrogen-fixing plants.
Dispersal syndrome and landscape fragmentation in the salt-marsh specialist spiderErigone longipalpis
Authors: Dahirel Maxime , Wullschleger Marie , Berry Tristan , Croci Solène , Pétillon Julien 
Published: December 2021

AbstractDispersal and its evolution play a key role for population persistence in fragmented landscapes where habitat loss and fragmentation increase the cost of between-habitat movements. In such contexts, it is important to know how variation in dispersal and other traits is structured, and whether responses to landscape fragmentation are aligned with underlying dispersal-trait correlations, or dispersal syndromes. We therefore studied trait variation inErigone longipalpis, a European spider species specialist of (often patchy) salt marshes. We collected spiders in two salt-marsh landscapes differing in habitat availability. We then reared lab-born spiders for two generations in controlled conditions, and measured dispersal and its association with various key traits.E. longipalpispopulation densities were lower in the more fragmented landscape. Despite this, we found no evidence of differences in dispersal, or any other trait we studied, between the two landscapes. While a dispersal syndrome was present at the among-individual level (dispersers were more fecund and faster growing, among others), there was no indication it was genetically driven: among-family differences in dispersal were not correlated with differences in other traits. Instead, we showed that the observed phenotypic covariations were mostly due to within-family correlations. We hypothesize that the dispersal syndrome is the result of asymmetric food access among siblings, leading to variation in development rates and carrying over to adult traits. Our results show we need to better understand the sources of dispersal variation and syndromes, especially when dispersal may evolve rapidly in response to environmental change.
Insertions in the SARS-CoV-2 Spike N-Terminal Domain May Aid COVID-19 Transmission
Authors: Lam Su Datt , Waman Vaishali P , Orengo Christine , Lees Jonathan 
Published: December 2021

AbstractCoronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 is an ongoing pandemic that causes significant health/socioeconomic burden. Variants of concern (VOCs) have emerged which may affect transmissibility, disease severity and re-infection risk. Most studies focus on the receptor-binding domain (RBD) of the Spike protein. However, some studies suggest that the Spike N-terminal domain (NTD) may have a role in facilitating virus entry via sialic-acid receptor binding. Furthermore, most VOCs include novel NTD variants. Recent analyses demonstrated that NTD insertions in VOCs tend to lie close to loop regions likely to be involved in binding sialic acids. We extended the structural characterisation of these putative sugar binding pockets and explored whether variants could enhance the binding to sialic acids and therefore to the host membrane, thereby contributing to increased transmissibility. We found that recent NTD insertions in VOCs (i.e., Gamma, Delta and Omicron variants) and emerging variants of interest (VOIs) (i.e., Iota, Lambda, Theta variants) frequently lie close to known and putative sugar-binding pockets. For some variants, including the recent Omicron VOC, we find increases in predicted sialic acid binding energy, compared to the original SARS-CoV-2, which may contribute to increased transmission. We examined the similarity of NTD across a range of related Betacoronaviruses to determine whether the putative sugar-binding pockets are sufficiently similar to be exploited in drug design. Despite global sequence and structure similarity, most sialic-acid binding pockets of NTD vary across related coronaviruses. Typically, SARS-CoV-2 possesses additional loops in these pockets that increase contact with polysaccharides. Our work suggests ongoing evolutionary tuning of the sugar-binding pockets in the virus. Whilst three of the pockets are too structurally variable to be amenable to pan Betacoronavirus drug design, we detected a fourth pocket that is highly structurally conserved and could therefore be investigated in pursuit of a generic drug. Our structure-based analyses help rationalise the effects of VOCs and provide hypotheses for experiments. For example, the Omicron variant, which has increased binding to sialic acids in pocket 3, has a rather unique insertion near pocket 3. Our work suggests a strong need for experimental monitoring of VOC changes in NTD.
Learning Active Multimodal Subspaces in the Brain
Authors: Batta Ishaan , Abrol Anees , Fu Zening , Calhoun Vince D. 
Published: December 2021

ABSTRACTHere we introduce a multimodal framework to identify subspaces in the human brain that are defined by collective changes in structural and functional measures and are actively linked to demographic, biological and cognitive indicators in a population. We determine the multimodal subspaces using principles of active subspace learning (ASL) and demonstrate its application on a sample learning task (biological ageing) on a Schizophrenia dataset. The proposed multimodal ASL method successfully identifies latent brain representations as subsets of brain regions and connections forming co-varying subspaces in association with biological age. We show that Schizophrenia is characterized by different subspace patterns compared to those in a cognitively normal brain. The multimodal features generated by projecting structural and functional MRI components onto these active subspaces perform better than several PCA-based transformations and equally well when compared to non-transformed features on the studied learning task. In essence, the proposed method successfully learns active brain subspaces associated with a specific brain condition but inferred from the brain imaging data along with the biological/cognitive traits of interest.
Floral evolution and pollinator diversification inHedychiumJ.Koenig (Zingiberaceae): one of Mr. Darwin’s tropical fantasies
Authors: Ashokan Ajith , Suksathan Piyakaset , Leong-Škorničková Jana , Newman Mark , Kress W. John , Gowda Vinita 
Published: December 2021

ABSTRACTPREMISEHedychiumJ.Koenig (ginger lilies: Zingiberaceae) is endemic to the Indo-Malayan Realm (IMR) and is known for its fragrant flowers. Two different pollination syndromes characterize the genus: diurnal or bird pollination and nocturnal or moth pollination systems. To date, no attempt has been undertaken to understand the evolution of floral traits in this genus.METHODSWe estimated ancestral character-states, phylogenetic signals, and character correlations for thirteen discrete and eight continuous floral traits representing 75% species diversity ofHedychium. Diversification rate estimation analyses were also employed to understand trait-dependent diversification in the genus.RESULTSInflorescence structure, cincinnus capacity, and curvature of floral tubes revealed strong phylogenetic dependence, whereas number of open flowers per inflorescence per day, color of the labellum, and exertion of the stigma characterized higher ecological effects. Diversification rate estimations suggested that the labellum width, floral tube length, and labellum color played a major role in the evolutionary diversification ofHedychium.CONCLUSIONSWe identified bract type and cincinnus capacity as synapomorphies forHedychium, while the island-specific clade III was characterized by slender cylindrical inflorescence, coiling of floral tubes, and longer bract to calyx ratio. The circum-Himalayan clade IV is the most speciose, derived, and with most variable floral traits. Although floral color and size lacked any association with pollinator-specific traits (moth and bird pollination), pale colored flowers were most common in the early diverging clades (clade I, II-el., and II-de.), indicating their ancestral nature, when compared to brightly colored flowers.
Decoupling SARS-CoV-2 ORF6 localization and interferon antagonism
Authors: Wong Hoi Tong , Cheung Victoria , Salamango Daniel J. 
Published: December 2021

ABSTRACTLike many pathogenic viruses, SARS-CoV-2 must overcome interferon (IFN)-mediated host defenses for infection establishment. To achieve this, SARS-CoV-2 deploys overlapping mechanisms to antagonize IFN production and signaling. The strongest IFN antagonist is the accessory protein ORF6, which localizes to multiple membranous compartments, including the nuclear envelope, where it directly binds the nuclear pore components Nup98-Rae1 to inhibit nuclear translocation of activated STAT1/IRF3 transcription factors. However, a direct cause-and-effect relationship between ORF6 localization and IFN antagonism has yet to be explored experimentally. Here, we use extensive mutagenesis studies to define the structural determinants required for steady-state localization and demonstrate that mis-localized ORF6 variants can still potently inhibit nuclear trafficking and IFN signaling. Additionally, expression of a peptide that mimics the ORF6/Nup98 interaction domain robustly inhibited nuclear trafficking. Furthermore, pharmacologic and mutational approaches combined to suggest that ORF6 is likely a peripheral-membrane protein, opposed to being a transmembrane protein as previously speculated. Thus, ORF6 localization and IFN antagonism are independent activities, which raises the possibility that ORF6 may have additional functions within membrane networks to enhance virus replication.
Escherichia coli antimicrobial resistance; Phenotypic and genotypic characterization
Authors: Elsharawy Nagwa Thabet , Al-Zahrani Hind A. A. , El-Waseif Amr A. 
Published: December 2021

AbstractImproper use of the antimicrobials as E. coli giving the microorganism multi-resistance against many antimicrobials by gene mutation on integrons, transposons and plasmids. Therefore, our aim in this study is to 1)examine antibiotics resistance phenotype and genotype in Escherichia coli, 2) identifying the structure of bacterial resistance genes on whole-genome sequencing against multi-drug resistant of Escherichia coli in marketed poultry meat. Samples collected, prepared and Bacteriological examination, Antimicrobial sensitivity test performed, Serological identification of Escherichia coli isolates. Results declared that; the prevalence of E. coli from tested chicken meat samples of 100 chicken meat samples surveyed against E. coli the result declared that about 40%. Antimicrobial susceptibility was; antibiotics of choice against E. coli Sulfonamides, Cephalosporins,Tetracyclines, Quinolones. Serologically, STEC (O157:H7) 30%, ETEC (O142) 10%, EHEC (O26:H11). The subunit B of shiga-like toxin (SLT) gene appeared as a homogenous band. Heat-labile toxin (LT) gene was screened in both genomic DNA and plasmid preps in tested strains. Control STEC as it represents a danger to the poultry consumers. We recommended to increase the hygienic measures during slaughtering, processing and/or handling of chicken carcasses and avoidance unnecessary usage of any antimicrobials to avoid appearance of new antimicrobials resistant.
BDNF-dependent modulation of axonal transport is selectively impaired in ALS
Authors: Tosolini Andrew P. , Sleigh James N. , Surana Sunaina , Rhymes Elena R. , Cahalan Stephen D. , Schiavo Giampietro 
Published: December 2021

AbstractAxonal transport ensures long-range delivery of essential cargoes between proximal and distal compartments of neurons, and is needed for neuronal development, function, and survival. Deficits in axonal transport have been detected at pre-symptomatic stages in mouse models of amyotrophic lateral sclerosis (ALS), suggesting that impairments are fundamental for disease pathogenesis. However, the precise mechanisms responsible for the transport deficits and whether they preferentially affect α-motor neuron (MN) subtypes remain unresolved. Here, we report that stimulation of wild-type neurons with brain-derived neurotrophic factor (BDNF) enhances trafficking of signalling endosomes specifically in fast MNs (FMNs). In early symptomatic SOD1G93Amice, FMNs display selective impairment of axonal transport and develop an insensitivity to BDNF stimulation, with pathology upregulating classical non-pro-survival receptors in muscles and sciatic nerves. Altogether, these data indicate that cell- and non-cell autonomous BDNF signalling is impaired in vulnerable SOD1G93AMNs, thus identifying a new key deficit in ALS amenable for future therapeutic interventions.
Slit-Robo Signalling Establishes a Sphingosine-1-Phosphate Gradient to Polarise Fin Mesenchyme and Establish Fin Morphology
Authors: Mahabaleshwar Harsha , Asharani P.V. , Yi Jun Tricia Loo , Koh Shze Yung , Pitman Melissa R. , Kwok Samuel , Ma Jiajia , Hu Bo , Lin Fang , Li Lok Xue , Pitson Stuart M. , Saunders Timothy E. , Carney Tom J. 
Published: December 2021

SUMMARYImmigration of mesenchymal cells into the growing fin and limb buds drives distal outgrowth, with subsequent tensile forces between these cells essential for fin and limb morphogenesis. Morphogens derived from the apical domain of the fin, orientate limb mesenchyme cell polarity, migration, division and adhesion. The zebrafish mutantstompdisplays defects in fin morphogenesis including blister formation and associated loss of orientation and adhesion of immigrating fin mesenchyme cells. Positional cloning ofstompidentified a mutation in the gene encoding the axon guidance ligand, Slit3. We provide evidence that Slit ligands derived from immigrating mesenchyme act via Robo receptors at the Apical Ectodermal Ridge (AER) to promote release of sphingosine-1-phosphate (S1P). S1P subsequently diffuses back to the mesenchyme to promote their polarisation, orientation, positioning and adhesion to the interstitial matrix of the fin fold. We thus demonstrate coordination of the Slit-Robo and S1P signalling pathways in fin fold morphogenesis. Our work introduces a mechanism regulating the orientation, positioning and adhesion of its constituent cells.
Seroprevalence of brucellosis and molecular characterization of Brucella spp. from slaughtered cattle in Rwanda
Authors: Ntivuguruzwa Jean Bosco , Kolo Francis Babaman , Mwikarago Emil Ivan , vanHeerden Henriette 
Published: December 2021

AbstractBovine brucellosis is endemic in Rwanda, although, there is paucity of documented evidence about the disease in slaughtered cattle. A cross-sectional study was conducted in slaughtered cattle (n=300) to determine the seroprevalence of anti-Brucella antibodies using the Rose Bengal Test (RBT), and indirect enzyme-linked immunosorbent assay (i-ELISA). Corresponding tissues were cultured onto a modified Centro de Investigación y Tecnología Agroalimentaria (CITA) selective medium and analysed for Brucella spp. using the 16S-23S ribosomal interspacer region (ITS), AMOS, and Bruce-ladder PCR assays. The RBT seroprevalence was 20.7% (62/300), and 2.9% (8/300) with i-ELISA and 2.9% (8/300) using both tests in parallel. Brucella specific 16S-23S ribosomal DNA interspace region (ITS) PCR detected Brucella DNA in 5.6% (17/300; Brucella culture prevalence). AMOS-PCR assay identified mixed B. abortus and B. melitensis (n=3), B. abortus (n=3) and B. melitensis (n=5) while Bruce-ladder PCR also identified B. abortus (n=5) and B. melitensis (n=6). The gold standard culture method combined with PCR confirmation identified 5.6% Brucella cultures which is higher than the more sensitive seroprevalence of 2.9%. This emphasizes the need to validate the serological tests in Rwanda. The mixed infection caused by B. abortus and B. melitensis in slaughtered cattle indicates cross-infection and poses a risk of exposure potential to abattoir workers. It is essential to urgently strengthen the national bovine brucellosis control program through vaccination as well as test- and-slaughter.