bluuebunny/crossref_metadata_2025_split huggingface dataset:
UnveilingCryptosporidium parvumSporozoite-Derived Extracellular Vesicles: Profiling, Origin, and Protein Composition
Authors: Bertuccini Lucia , Boussadia Zaira , Salzano Anna Maria , Vanni Ilaria , Passerò Ilaria , Nocita Emanuela , Scaloni Andrea , Sanchez Massimo , Sargiacomo Massimo , Fiani Maria Luisa , Tosini Fabio 
Published: December 2023

AbstractCryptosporidium parvumis a common cause of a zoonotic disease and a main cause of diarrhea in newborns around the world. Effective drugs or vaccines are still lacking. Oocyst is the infective form of the parasite; after the ingestion the oocyst excysts and releases four sporozoites into the intestine that rapidly attack the enterocytes. The membrane protein CpRom1 is a large rhomboid protease that is expressed by sporozoites and recognized as antigen by the host immune system. In this study, we observed the release of CpRom1 with extracellular vesicles (EVs) not previously described. To investigate this phenomenon, we isolated and resolved EVs from the excystation medium by differential ultracentrifugation. By fluorescence flow cytometry and transmission electron microscopy (TEM), we identified two types of sporozoite-derived vesicles: large extracellular vesicles (LEVs) and small extracellular vesicles (SEVs), having different a mean size of 150 nm and 60 nm, respectively. Immunodetection experiments proved the occurrence of CpRom1 and the Golgi protein CpGRASP in LEVs, while immune-electron microscopy experiments demonstrated localization of CpRom1 on LEVs surface.TEM and scanning electron microscopy (SEM) showed the generation of LEVs by budding of the outer membrane of sporozoites; conversely, the origin of SEVs remained uncertain.Differences between LEVs and SEVs were observed for protein composition as proved by the corresponding electrophoretic profiles. Indeed, a dedicated proteomic analysis identified 5 proteins unique to LEVs composition and 16 proteins unique to SEVs. Overall, 60 proteins were identified in the proteome of both types of vesicles and most of these proteins (48 in number) were already identified in the molecular cargo of extracellular vesicles from other organisms. Noteworthy, we identified 12 proteins unique toCryptosporidiumspp. that had never been associated with EVs. This last group included the immunodominant parasite antigen glycoprotein GP60, which is one of the most abundant proteins in both LEVs and SEVs.
Structure and methyl-lysine binding selectivity of the HUSH complex subunit MPP8
Authors: Nikolopoulos Nikos , Oda Shun-ichiro , Prigozhin Daniil M. , Modis Yorgo 
Published: December 2023

AbstractThe Human Silencing Hub (HUSH) guards the genome from the pathogenic effects of retroelement expression. Composed of MPP8, TASOR, and Periphilin-1, HUSH recognizes actively transcribed retrotransposed sequences by the presence of long (>1.5-kb) nascent transcripts without introns. HUSH recruits effectors that alter chromatin structure, degrade transcripts, and deposit transcriptionally repressive epigenetic marks. Here, we report the crystal structure of the C-terminal domain (CTD) of MPP8 necessary for HUSH activity. The MPP8 CTD consists of five ankyrin repeats followed by a domain with structural homology to the PINIT domains of Siz/PIAS-family SUMO E3 ligases. AlphaFold-Multimer modeling predicts that the MPP8 CTD forms extended interaction interfaces with a SPOC domain and a domain with a novel fold in TASOR. The MPP8 chromodomain, known to bind the repressive mark H3K9me3, binds with similar or higher affinity to sequences in the H3K9 methyltransferase subunits SETDB1, ATF7IP, G9a, and GLP. Hence, MPP8 promotes heterochromatinization by recruiting H3K9 methyltransferases. Our work identifies novel structural elements in MPP8 required for HUSH complex assembly and silencing, thereby fulfilling vital functions in controlling retrotransposons.
Shared features of blastula and neural crest stem cells evolved at the base of vertebrates
Authors: York Joshua R. , Rao Anjali , Huber Paul B. , Schock Elizabeth N. , Montequin Andrew , Rigney Sara , LaBonne Carole 
Published: December 2023

AbstractThe neural crest is vertebrate-specific stem cell population that helped drive the origin and evolution of the vertebrate clade. A distinguishing feature of these stem cells is their multi-germ layer potential, which has drawn developmental and evolutionary parallels to another stem cell population—pluripotent embryonic stem cells (animal pole cells or ES cells) of the vertebrate blastula. Here, we investigate the evolutionary origins of neural crest potential by comparing neural crest and pluripotency gene regulatory networks (GRNs) in both jawed (Xenopus) and jawless (lamprey) vertebrates. Through comparative gene expression analysis and transcriptomics, we reveal an ancient evolutionary origin of shared regulatory factors between neural crest and pluripotency GRNs that dates back to the last common ancestor of extant vertebrates. Focusing on the key pluripotency factorpou5(formerly oct4), we show that the lamprey genome encodes apou5ortholog that is expressed in animal pole cells, as in jawed vertebrates, but is absent from the neural crest. However, gain-of-function experiments show that both lamprey andXenopus pou5enhance neural crest formation, suggesting thatpou5was lost from the neural crest of jawless vertebrates. Finally, we show thatpou5is required for neural crest specification in jawed vertebrates and that it acquired novel neural crest-enhancing activity after evolving from an ancestralpou3-like clade that lacks this functionality. We propose that a pluripotency-neural crest GRN was assembled in stem vertebrates and that the multi-germ layer potential of the neural crest evolved by deploying this regulatory program.
WITHDRAWN: (CA)noligonucleotides correct RNA mis-splicing in TDP-43 pathology
Authors: Wang Congwei , Terrigno Marco , Joenson Lars , Li Meiling , Schröter Manuel , Nordbo Bettina , Chyzynska Katarzyna , Weile Christian , Bastien Jessica , Mogensen Marianne B. , Seidel Antonia S. , Hagedorn Peter H. , Jagasia Ravi , Vikesaa Jonas 
Published: December 2023

Withdrawal StatementThe authors have withdrawn their manuscript owing to not all authors having approved the manuscript. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author.
Deep super-resolution imaging of thick tissue using structured illumination with adaptive optics
Authors: Wang Jingyu , Stoychev Danail , Phillips Mick , Pinto David Miguel Susano , Parton Richard M. , Hall Nick , Titlow Josh , Faria Ana Rita , Wincott Matthew , Gala Dalia , Gerondopoulos Andreas , Irani Niloufer , Dobbie Ian , Schermelleh Lothar , Booth Martin , Davis Ilan 
Published: December 2023

Abstract3D structured illumination microscopy (3D-SIM) doubles the resolution of fluorescence imaging in lateral and axial directions and increases contrast in both fixed and live specimens. However, 3D-SIM has so far not been widely applied to imaging deep in thick tissues due to its sensitivity to specimen-induced aberrations, making the method difficult to apply beyond 10 µm in depth. Furthermore, 3D-SIM has not been available in an upright configuration, limiting its use for live imaging while manipulating the specimen, for example with electrophysiology. Here, we have overcome these barriers by developing a novel upright 3D-SIM system (termed Deep3DSIM) that incorporates adaptive optics (AO) for aberration correction and remote focusing, reducing artefacts, and removing the need to move the specimen or objective. Both these advantages are equally applicable to inverted 3D-SIM microscopes. We demonstrate high-quality 3D-SIM imaging up to 130 µm into complex tissue and live sample manipulation, including human cells andDrosophilalarval brains and embryos.
Porcine sapovirus protease controls the innate immune response and targets TBK1
Authors: Georgana Iliana , Hosmillo Myra , Jahun Aminu S. , Emmott Edward , Sorgeloos Frederic , Cho Kyoung-Oh , Goodfellow Ian G. 
Published: December 2023

AbstractHuman sapoviruses (HuSaVs) and noroviruses are considered the leading cause of acute gastroenteritis worldwide. While extensive research has focused on noroviruses, our understanding of sapoviruses (SaVs) and their interactions with the host’s immune response remains limited. HuSaVs have been challenging to propagate in vitro, making the porcine sapovirus (PSaV) Cowden strain a valuable model for studying SaV pathogenesis. In this study we show, for the first time, that PSaV Cowden strain has mechanisms to evade the host’s innate immune response. The virus 3C-like protease (NS6) inhibits type I IFN production by targeting TBK1. Catalytically active NS6, both during ectopic expression and during PSaV infection, targets TBK1 which is then led for rapid degradation by the proteasome. Moreover, deletion of TBK1 from porcine cells led to a significant increase in PSaV titres, emphasizing its role in regulating PSaV infection. Additionally, we successfully established PSaV infection in IPEC-J2 cells, an enterocytic cell line originating from the jejunum of a neonatal piglet. Overall, this study provides novel insights into PSaV evasion strategies, opening the way for future investigations for SaV-host interactions, and enabling the use of a new cell line model for PSaV research.
DHCR24-mediated sterol homeostasis during spermatogenesis is required for sperm mitochondrial sheath formation and impacts male fertility over time
Authors: Relovska Sona , Wang Huafeng , Zhang Xinbo , Fernández-Tussy Pablo , Jeong Kyung Jo , Choi Jungmin , Suárez Yajaira , McDonald Jeffrey G. , Fernández-Hernando Carlos , Chung Jean-Ju 
Published: December 2023

AbstractDesmosterol and cholesterol are essential lipid components of the sperm plasma membrane. Cholesterol efflux is required for capacitation, a process through which sperm acquire fertilizing ability. In this study, using a transgenic mouse model overexpressing 24-dehydrocholesterol reductase (DHCR24), an enzyme in the sterol biosynthesis pathway responsible for the conversion of desmosterol to cholesterol, we show that disruption of sterol homeostasis during spermatogenesis led to defective sperm morphology characterized by incomplete mitochondrial packing in the midpiece, reduced sperm count and motility, and a decline in male fertility with increasing paternal age, without changes in body fat composition. Sperm depleted of desmosterol exhibit inefficiency in the acrosome reaction, metabolic dysfunction, and an inability to fertilize the egg. These findings provide molecular insights into sterol homeostasis for sperm capacitation and its impact on male fertility.
mDia2 is an important mediator of MRTF-A-dependent regulation of breast cancer cell migration
Authors: Eder Ian , Yu Virginia , Antonello Jacob , Chen Fangyuan , Gau David , Chawla Pooja , Joy Marion , Lucas Peter , Boone David , Lee Adrian V. , Roy Partha 
Published: December 2023

ABSTRACTDysregulated actin cytoskeleton gives rise to aberrant cell motility and metastatic spread of tumor cells. This study evaluates the effect of overexpression of wild-type vs functional mutants of MRTF-A on migration and invasion of breast cancer (BC) cells. Our studies indicate that SRF’s interaction is critical for MRTF-A-induced promotion of both 2D and 3D cell migration, while the SAP-domain function is important selectively for 3D cell migration. Increased MRTF-A activity is associated with more effective membrane protrusion, a phenotype that is attributed predominantly to SRF’s interaction of MRTF. We demonstrate formin-family protein mDia2 as an important mediator of MRTF-stimulated actin polymerization at the leading edge and cell migration. Multiplexed quantitative immunohistochemistry and transcriptome analyses of clinical BC specimens further demonstrate a positive correlation between nuclear localization of MRTF with malignant traits of cancer cells and enrichment of MRTF-SRF gene signature in pair-matched distant metastases vs primary tumors. In conclusion, this study establishes a novel mechanism of MRTF-dependent regulation of cell migration and provides evidence for the association between MRTF activity and increased malignancy in human breast cancer, justifying future development of a specific small molecule inhibitor of the MRTF-SRF transcriptional complex as a potential therapeutic agent in breast cancer.SIGNIFICANCEActin cytoskeletal dysregulation gives rise to metastatic dissemination of cancer cells. This study mechanistically investigates the impact of specific functional disruption of MRTF (a transcriptional co-factor of SRF) on breast cancer cell migration.This study establishes a novel mechanism linking mDia2 to MRTF-dependent regulation of cell migration and provides clinical evidence for the association between MRTF activity and increased malignancy in human breast cancer.Findings from these studies justify future exploration of specific small molecule inhibitor of the MRTF-SRF transcriptional complex as a potential therapeutic agent in breast cancer.
Intracellular pH modulates vimentin remodeling in response to oxidants
Authors: Martínez Alma E. , González-Jiménez Patricia , Vidal-Verdú Cristina , Pajares María A. , Pérez-Sala Dolores 
Published: December 2023

AbstractThe intermediate filament protein vimentin plays key roles in the integration of cytoskeletal functions with impact on essential cellular processes including migration, mitosis and autophagy. Moreover, vimentin is involved in pathological processes such as cancer, fibrosis and interaction with pathogens. The vimentin network is finely tuned by posttranslational modifications, among which, those affecting its single cysteine residue (C328) play a critical role not only in vimentin function, but also in its interplay with actin. Interestingly, C328 exhibits a low pKa, which favors the presence of the thiol group in the thiolate form, and therefore its reactivity, at physiological pH. Therefore, C328 reactivity and modifications could be modulated by pH fluctuations in the physiological range. Here we show that indeed, vimentin cysteine oxidation and alkylation, and the subsequent vimentin remodeling, can be modulated as a function of pH, in vitro and in cells. Lowering intracellular pH by several means renders vimentin unresponsive to disruption by oxidants, whereas provoking an intracellular alkalinization exerts a sensitizing effect. The protective effect of low pH appears selective for vimentin since it does not preclude oxidant-elicited disruption of actin or tubulin structures. Importantly, a C328A vimentin mutant is resistant to disruption by oxidants under all pH conditions, highlighting the role of the thiol group at this position in the pH-dependent modulation of vimentin susceptibility to oxidants. Taken together, these results put forward intracellular pH as a key factor modulating redox-dependent vimentin remodeling.
Lamin B Receptor Upregulation in Metastatic Melanoma Causes Cholesterol-Mediated Nuclear Envelope Fragility
Authors: Baird Michelle A. , Fischer Robert S. , Jewett Cayla E. , Malide Daniela , Cartagena-Rivera Alexander X. , Waterman Clare M. 
Published: December 2023

AbstractMetastatic cancer cells migrate through regions of tissue confinement, causing nuclear envelope (NE) rupture and heritable DNA damage. We discovered that cells from multiple cancers have increased NE fragility in confinement and transcriptional upregulation of nuclear genes compared to benign counterparts. A bioinformatic-driven siRNA screen revealed that lamin B receptor (LBR) upregulation correlates with melanoma progression and NE fragility. Increased LBR cholesterol synthase activity causes accumulation of cholesterol in the NE, which is necessary and sufficient for nuclear deformability and NE rupture in cells confinedin vitroand is associated with NE rupture in invasive cells migrating out of tumor organoids and tumorsin vivo.Thus, LBR upregulation causes excess NE cholesterol, driving nuclear fragility in confined migrating melanoma cells, establishing a direct role for nuclear membrane lipid composition in metastatic cancers.
The Reelin Receptor ApoER2 is a Cargo for the Adaptor Protein Complex AP-4: Implications for Hereditary Spastic Paraplegia
Authors: Caracci Mario O. , Pizarro Héctor , Alarcón-Godoy Carlos , Fuentealba Luz M. , Farfán Pamela , Pace Raffaella De , Santibañez Natacha , Cavieres Viviana A. , Pástor Tammy P. , Bonifacino Juan S. , Mardones Gonzalo A. , Marzolo María-Paz 
Published: December 2023

ABSTRACTAdaptor protein complex 4 (AP-4) is a heterotetrameric complex that promotes protein export from thetrans-Golgi network. Mutations in each of the AP-4 subunits cause a complicated form of Hereditary Spastic Paraplegia (HSP). Herein, we report that ApoER2, a receptor in the Reelin signaling pathway, is a cargo of the AP-4 complex. We identify the motif ISSF/Y within the ApoER2 cytosolic domain as necessary for interaction with the canonical signal-binding pocket of the µ4 (AP4M1) subunit of AP-4.AP4E1-knock-out (KO) HeLa cells and hippocampal neurons fromAp4e1-KO mice display increased Golgi localization of ApoER2. Furthermore, hippocampal neurons fromAp4e1-KO mice andAP4M1-KO human iPSC-derived cortical i3Neurons exhibit reduced ApoER2 protein expression. Analyses of biosynthetic transport of ApoER2 reveal differential post-Golgi trafficking of the receptor, with lower axonal distribution in KO compared to wild-type neurons, indicating a role of AP-4 and the ISSF/Y motif in the axonal localization of ApoER2. Finally, analyses of Reelin signaling in mouse hippocampal and human cortical KO neurons show that AP4 deficiency causes no changes in Reelin-dependent activation of the AKT pathway and only mild changes in Reelin-induced dendritic arborization, but reduces Reelin-induced ERK phosphorylation, CREB activation, and Golgi deployment. Altogether, this work establishes ApoER2 as a novel cargo of the AP-4 complex, suggesting that defects in the trafficking of this receptor and in the Reelin signaling pathway could contribute to the pathogenesis of HSP caused by mutations in AP-4 subunits.
Nerve Growth Factor Receptor Limits Inflammation to Promote Remodeling and Repair of Osteoarthritic Joints
Authors: Zhao Lan , Lai Yumei , Jiao Hongli , Huang Jian 
Published: December 2023

AbstractOsteoarthritis (OA) is a painful, incurable disease affecting over 500 million people. The need for relieving OA pain is paramount but inadequately addressed, partly due to limited understandings of how pain signaling regulates non-neural tissues. Here we report that nerve growth factor receptor (NGFR) is upregulated in skeletal cells during OA and plays an essential role in the remodeling and repair of osteoarthritic joints. Specifically, NGFR is expressed in osteochondral cells but not in skeletal progenitor cells and induced by TNFα to attenuate NF-κB activation, maintaining proper BMP-SMAD1 signaling and suppressing RANKL expression. NGFR deficiency hyper-activates NF-κB in murine osteoarthritic joints, which impairs bone formation and enhances bone resorption as exemplified by a reduction in subchondral bone and osteophytes. In human OA cartilage, NGFR is also negatively associated with NF-κB activation. Together, this study uncovers a role of NGFR in limiting inflammation for repair of diseased skeletal tissues.
Toward digitalization strategic perspective in the European food industry: non-linear nexuses analysis
Authors: Alnoor Alhamzah , Atiyah Abbas Gatea , Abbas Sammar 
Published: December 2023

Purpose Organizations deal with digital technologies to achieve their strategic goals. The shift toward digitization is a major challenge because it requires companies to create a digital outlook that influences organizational design. As a result, investigation of institutional theory and entrepreneurial orientation theory in the European food industry has become the focus of research in recent times.Design/methodology/approach To this end, data were collected from 83 companies related to the food industry in the European context. By applying a hybrid phase of the partial least squares structural equation modeling (PLS-SEM) and artificial neural network (ANN) methods, this study captured the causal–non-linear relationships among the study constructs.Findings The findings revealed that the variables of institutional theory and entrepreneurial orientation theory affect the adoption of the digital strategy. There is also a dual interaction role for e-business capabilities and digital transformation. The results of non-linear relationships confirmed that digital strategy adoption is highly influenced by digital transformation, followed by risk-taking, digital leadership, e-business capabilities, organizational agility, proactiveness and innovativeness.Research limitations/implications The authors provided significant implications for practitioners and academics about the most influential determinants of digital strategy – businesses must move swiftly toward digitization across its various units to achieve their objectives. An organization’s leadership must realize that equipping the employees with necessary skills is the first step toward digitalization.Originality/value The current study underscores the digital strategy, which is usually an overlooked area of investigation, in the food industry. The study identifies some important predictors of digital strategy adoption with the interaction’s role of digital transformation and e-business capabilities. Such relationships have been rarely discussed. In addition, the adoption of a hybrid SEM-AAN approach makes the study an original one.
Challenges of the MSE sector in the digital economy in Poland and Ukraine: comparative and statistical analysis
Authors: Remlein Marzena , Chugaievska Svitlana , Dehnel Grażyna , Romanchuk Kateryna 
Published: December 2023

PurposeThe authors aimed to examine how the level of digitalization in Poland and Ukraine affects the contribution of small and medium-sized enterprises (SMEs) to the countries’ gross domestic product (GDP).Design/methodology/approachThe study involved a comparative analysis and statistical modeling of the impact of key economic factors on the contribution of SMEs to Poland’s and Ukraine’s GDP in the 2010–2020 period. The authors used principles of the theory of economic growth and calculated the coefficient of digital competitiveness as a composite indicator consisting of a number of global indices.FindingsThe study revealed significant differences between both countries, which can be attributed to a higher level of digitalization in Polish SMEs. The authors used the Polish experience to recommend how to reform Ukraine’s digital economy in postwar recovery.Originality/valueThe contribution of SMEs to Poland’s GDP is higher than that of Ukraine’s because of the higher entrepreneurship rate in the Polish micro and small enterprises (MSEs) sector. The authors found that a unit change in the integrated coefficient of digital competitiveness is related to the greatest change in the contribution of SMEs to the country’s GDP when the other factors in the model equation remain fixed.