bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ultrasound-mediated drug diffusion, uptake, and cytotoxicity in glioblastoma 3D tumour sphere model
Authors: Wanigasekara Janith , Mondala Julie Rose Mae , Cullen Patrick J. , Tiwari Brijesh , Curtin James F. 
Published: October 2023

AbstractA myriad of biological effects can be stimulated by the ultrasound (US) for the treatment of cancer. The objective of our research was to investigate the effect of ultrasound alone and in combination with the chemotherapeutic drugs such as doxorubicin (DOX) and temozolomide (TMZ) on human glioblastoma (GBM) and the human epidermoid carcinoma cancer 2D and 3D cell cultures. Results indicated that the US 96-probe device could induce tumour sphere cytotoxicity in a dosage- and time-dependent manner, with multiple treatments augmenting this cytotoxic effect. With enhanced cytotoxicity, US decreased tumour sphere growth metabolic activity, disrupted spheroid integrity, and heightened the occurrence of DNA double strand breaks, resulting in damage to tumour spheres and an inability to rebuild tumour spheres after multiple US treatments. The combination of US and TMZ / DOX enhanced the efficiency of treatment for GBM and epidermoid carcinoma by enhancing induced cytotoxicity in 3D tumour spheres compared to 2D monolayer cells, and also by increasing the incubation time, which is the most crucial way to differentiate between the effectiveness of drug treatment with and without US. In conclusion, our data demonstrate that US enhances drug diffusion, uptake, and cytotoxicity using 3D spheroid models when compared with 2D cultures. It also demonstrates the significance of 3D cell culture models in drug delivery and discovery research.Graphical Abstract
Identification of visible and near-infrared signature peaks for arboviruses andPlasmodium
Authors: Goh Brendon , Soares Magalhães Ricardo J. , Ciocchetta Silvia , Liu Wenjun , Sikulu-Lord Maggy T. 
Published: October 2023

AbstractArbovirus and malaria infections affect more than half of the world’s population causing major financial and physical burden. Current diagnostic tools such as microscopy, molecular and serological techniques are technically demanding, costly, or time consuming. Near-infrared spectroscopy has recently been demonstrated as a potential diagnostic tool for malaria and arbovirus and as a screening tool for disease vectors. However, pathogen specific infrared peaks that allow detection of these infections are yet to be described. In this study, we identified unique NIRS peaks from existing laboratory strains of four major arboviruses including Barmah Forest virus (BFV), Dengue virus (DENV), Ross River virus (RRV), Sindbis virus (SINV) andPlasmodium falciparum. Secondly, to determine the diagnostic ability of these peaks, we developed machine learning algorithms using Artificial Neural network (ANN) to differentiate arboviruses from media in which they are grown. Signature peaks for BFV were identified within the visible region at 410, 430, 562 and 588nm and the NIR region at, 946, 958, 1130, 1154 and 1780 nm. DENV related peaks were seen at 410nm within the visible region and 1130 nm within the NIR region. Signature peaks for RRV were observed within the visible region at 410 and 430 nm and within the NIR region at 1130 and 1780 nm, while SINV had a prominent peak at 410 nm within the visible region. Peaks at 514, 528, 547, 561, 582, and 595nm and peaks at 1388, 1432, 1681, 1700, 1721, 1882, 1905, 2245, 2278, 2300 nm were unique forP. falciparum. NIRS predictive sensitivity defined as the ability to predict an arbovirus as an infection was 90% (n = 20) for BFV, 100% (n =10) for RRV and 97.5% (n= 40) for DENV, while infection specificity defined as the ability to predict media as not-infected was 100% (n= 10). Our findings indicate that spectral signatures obtained by NIRS are potential biomarkers for diagnosis arboviruses and malaria.Author summaryMore than half of the world is at risk of contracting either malaria or arboviral infections. In resource limited settings, timely detection of these infections and the ability to screen thousands of people in a day using affordable tools is key to preventing their spread and unprecedented outbreaks. This emphasizes the need to develop portable, rapid, and easy to use diagnostic and surveillance tools. The near-infrared spectroscopy technique has recently been shown to be a potential tool for the diagnosis and surveillance of both malaria and arboviral infections. However, signature spectral biomarkers for these infections remain to be described. This study has identified several spectral biomarkers for DEN, RRV, BVF, SIN arboviruses andP. falciparum parasites. These biomarkers will assist the future assessment of this technique as a diagnostic and or surveillance tool for these infections in the field.
Mechanistic models for West Nile Virus transmission: A systematic review of features, aims, and parameterisation
Authors: de Wit Mariken , Dimas Martins Afonso , Delecroix Clara , Heesterbeek Hans , ten Bosch Quirine 
Published: October 2023

ABSTRACTMathematical models within the Ross-Macdonald framework increasingly play a role in our understanding of vector-borne disease dynamics and as tools for assessing scenarios to respond to emerging threats. These threats are typically characterised by a high degree of heterogeneity, introducing a range of possible complexities in models and challenges to maintain the link with empirical evidence.We systematically identified and analysed a total of 67 published papers presenting compartmental West Nile Virus (WNV) models that use parameter values derived from empirical studies. Using a set of fifteen criteria, we measured the dissimilarity compared to the Ross–Macdonald framework. We also retrieved the purpose and type of models and traced the empirical sources of their parameters.Our review highlights the increasing refinements in WNV models. Models for prediction included the highest number of refinements. We found uneven distributions of refinements and of evidence for parameter values. We identified several challenges in parameterising such increasingly complex models. For parameters common to most models, we also synthesise the empirical evidence for their values and ranges. The study highlights the potential to improve the quality of WNV models and their applicability for policy by establishing closer collaboration between mathematical modelling and empirical work.
Immune Epitopes of SARS-CoV-2 Spike Protein and Considerations for Universal Vaccine Development
Authors: Magazine Nicholas , Zhang Tianyi , Bungwon Anang D. , McGee Michael C. , Wu Yingying , Veggiani Gianluca , Huang Weishan 
Published: October 2023

AbstractDespite the success of global vaccination programs in slowing the spread of COVID-19, these efforts have been hindered by the emergence of new SARS-CoV-2 strains capable of evading prior immunity. The mutation and evolution of SARS-CoV-2 have created a demand for persistent efforts in vaccine development. SARS-CoV-2 Spike protein has been the primary target for COVID-19 vaccine development, but it is also the hotspot of mutations directly involved in host susceptibility and immune evasion. Our ability to predict emerging mutants and select conserved epitopes is critical for the development of a broadly neutralizing therapy or a universal vaccine. In this article, we review the general paradigm of immune responses to COVID-19 vaccines, highlighting the immunological epitopes of Spike protein that are likely associated with eliciting protective immunity resulting from vaccination. Specifically, we analyze the structural and evolutionary characteristics of the SARS-CoV-2 Spike protein related to immune activation and function via the toll-like receptors (TLRs), B cells, and T cells. We aim to provide a comprehensive analysis of immune epitopes of Spike protein, thereby contributing to the development of new strategies for broad neutralization or universal vaccination.
Designer Fat Cells: Adipogenic Differentiation of CRISPR-Cas9 Genome-Engineered Induced Pluripotent Stem Cells
Authors: Ely E. V. , Kapinski A. T. , Paradi S. G. , Tang R. , Guilak F. , Collins K. H. 
Published: October 2023

AbstractAdipose tissue is an active endocrine organ that can signal bidirectionally to many tissues and organ systems in the body. With obesity, adipose tissue is a source of low-level inflammation that contributes to various co-morbidities and damage to downstream effector tissues. The ability to synthesize genetically engineered adipose tissue could have critical applications in studying adipokine signaling and the use of adipose tissue for novel therapeutic strategies. This study aimed to develop a method for non-viral adipogenic differentiation of genome-edited murine induced pluripotent stem cells (iPSCs) and to test the ability of such cells to engraft in micein vivo. Designer adipocytes were created from iPSCs, which can be readily genetically engineered using CRISPR-Cas9 to knock out or insert individual genes of interest. As a model system for adipocyte-based drug delivery, an existing iPSC cell line that transcribes interleukin 1 receptor antagonist under the endogenous macrophage chemoattractant protein-1 promoter was tested for adipogenic capabilities under these same differentiation conditions. To understand the role of various adipocyte subtypes and their impact on health and disease, an efficient method was devised for inducing browning and whitening of IPSC-derived adipocytes in culture. Finally, to study the downstream effects of designer adipocytesin vivo, we transplanted the designer adipocytes into fat-free lipodystrophic mice as a model system for studying adipose signaling in different models of disease or repair. This novel translational tissue engineering and regenerative medicine platform provides an innovative approach to studying the role of adipose interorgan communication in various conditions.
Viral genomic features predictorthopoxvirusreservoir hosts
Authors: Tseng Katie K. , Koehler Heather , Becker Daniel J. , Gibb Rory , Carlson Colin J. , del Pilar Fernandez Maria , Seifert Stephanie N. 
Published: October 2023

AbstractOrthopoxviruses (OPVs), including the causative agents of smallpox and mpox have led to devastating outbreaks in human populations worldwide. However, the discontinuation of smallpox vaccination, which also provides cross-protection against related OPVs, has diminished global immunity to OPVs more broadly. We apply machine learning models incorporating both host ecological and viral genomic features to predict likely reservoirs of OPVs. We demonstrate that incorporating viral genomic features in addition to host ecological traits enhanced the accuracy of potential OPV host predictions, highlighting the importance of host-virus molecular interactions in predicting potential host species. We identify hotspots for geographic regions rich with potential OPV hosts in parts of southeast Asia, equatorial Africa, and the Amazon, revealing high overlap between regions predicted to have a high number of potential OPV host species and those with the lowest smallpox vaccination coverage, indicating a heightened risk for the emergence or establishment of zoonotic OPVs. Our findings can be used to target wildlife surveillance, particularly related to concerns about mpox establishment beyond its historical range.
Fructose-2,6-bisphosphate restores DNA repair activity of PNKP and ameliorates neurodegenerative symptoms in Huntington’s disease
Authors: Chakraborty Anirban , Sreenivasmurthy Sravan Gopalkrishnashetty , Miller Wyatt , Huai Weihan , Biswas Tapan , Mandal Santi Mohan , Boscá Lisardo , Krishnan Balaji , Ghosh Gourisankar , Hazra Tapas 
Published: October 2023

AbstractHuntington’s disease (HD) and spinocerebellar ataxia type 3 (SCA3) are the two most prevalent polyglutamine (polyQ) neurodegenerative diseases, caused by CAG (encoding glutamine) repeat expansion in the coding region of the huntingtin (HTT) and ataxin-3 (ATXN3) proteins, respectively. We have earlier reported that the activity, but not the protein level, of an essential DNA repair enzyme, polynucleotide kinase 3’-phosphatase (PNKP), is severely abrogated in both HD and SCA3 resulting in accumulation of double-strand breaks in patients’ brain genome. While investigating the mechanistic basis for the loss of PNKP activity and accumulation of DNA double-strand breaks leading to neuronal death, we observed that PNKP interacts with the nuclear isoform of 6-phosphofructo-2-kinase fructose-2,6-bisphosphatase 3 (PFKFB3). Depletion of PFKFB3 markedly abrogates PNKP activity without changing its protein level. Notably, the levels of both PFKFB3 and its product fructose-2,6 bisphosphate (F2,6BP), an allosteric modulator of glycolysis, are significantly lower in the nuclear extracts of post-mortem brain tissues of HD and SCA3 patients. Supplementation of F2,6BP restored PNKP activity in the nuclear extracts of patients’ brain. Moreover, intracellular delivery of F2,6BP restored both the activity of PNKP and the integrity of transcribed genome in neuronal cells derived from striatum of HD mouse. Importantly, supplementing F2,6BP rescued the HD phenotype in Drosophila, suggesting F2,6BP to serve in vivo as a cofactor for the proper functionality of PNKP and thereby, of brain health. Our results thus provide a compelling rationale for exploring the therapeutic use of F2,6BP and structurally related compounds for treating polyQ diseases.SignificanceTo unravel the biological basis for the loss of PNKP activity in HD and SCA3, the two most prevalent polyglutamine neurodegenerative disorders, we analyzed PNKP interactome and found that the nuclear isoform of a glycolytic enzyme PFKFB3 associated with PNKP and other repair proteins forming a multiprotein complex. Surprisingly, we found that PFKFB3 and its biosynthetic product, F2,6BP are significantly low in the affected region of patients’ brain. Exogenous addition of F2,6BP restored PNKP activity in patients’ brain nuclear extract. Moreover, supplementing F2,6BP in HD cells and fruit flies restored genome integrity and rescued the disease symptoms. While there is no curative therapy for HD/SCA3, except symptom management, our discovery suggests that F2,6BP supplementation would be a promising therapeutic option.
A fully sequenced collection of homozygous EMS mutants for forward and reverse genetic screens inArabidopsis thaliana
Authors: Carrère Sébastien , Routaboul Jean-Marc , Savourat Pauline , Bellenot Caroline , López Hernán , Monnens Thomas Quiroz , Ricou Anthony , Camilleri Christine , Laufs Patrick , Mercier Raphael , Noël Laurent D. 
Published: October 2023

AbstractGenetic screens are powerful tools for biological research and are one of the reasons for the success of the thale cressArabidopsis thalianaas a model species. Here, we describe the whole-genome sequencing of 871 Arabidopsis lines from the Homozygous EMS Mutant (HEM) collection as a novel resource for forward and reverse genetics. With an average 576 high-confidence mutations per HEM line, over three independent mutations altering protein sequence are found on average per gene in the collection. Pilot reverse genetics experiments on reproductive, developmental and physiological traits confirmed the efficacy of the tool for identifying both null and knockdown alleles. The possibility of conducting subtle repeated phenotyping of HEM lines and the immediate availability of the mutations will empower forward genetic approaches. The sequence resource is searchable with the ATHEM web interface (https://lipm-browsers.toulouse.inra.fr/pub/ATHEM/), and the biological material is distributed by the Versailles Arabidopsis Stock Center.
The human developing cerebral cortex is characterized by an increased de novo expression of lncRNAs in excitatory neurons
Authors: Morales-Vicente David A. , Tahira Ana C. , Woellner-Santos Daisy , Amaral Murilo S. , Berzoti-Coelho Maria G. , Verjovski-Almeida Sergio 
Published: October 2023

AbstractBackgroundOutstanding human cognitive abilities are computed in the cerebral cortex, a mammalian-specific brain region and the place of massive biological innovation. Long noncoding RNAs (lncRNAs) have emerged as gene regulatory elements with higher evolutionary turnover than mRNAs. The many lncRNAs identified in neural tissues make them candidates for molecular sources of cerebral cortex evolution and disease. Here, we characterized the genomic and cellular shifts that occurred during the evolution of the lncRNA repertoire expressed in the developing cerebral cortex of humans and explored their role in the evolution of this brain region.ResultsUsing systems biology approaches and comparative transcriptomics, we comprehensively annotated the cortical transcriptomes of humans, macaques, mice, and chickens and classified human cortical lncRNAs into evolutionary groups as a function of their predicted minimal ages. LncRNA evolutionary groups showed differences in expression levels, splicing efficiencies, transposable element contents, genomic distributions, and transcription factor binding to their promoters. Furthermore, older lncRNAs showed preferential expression in germinative zones, outer radial glial cells, and cortical inhibitory neurons. In comparison, younger lncRNAs showed preferential expression in cortical excitatory neurons, belonged to human-specific gene coexpression modules, and were dysregulated in autism spectrum disorder.ConclusionsThese results suggest a shift in the roles of cortical lncRNAs over evolution, highlighting the antique lncRNAs as a source of molecular evolution of conserved developmental programs; conversely, thede novoexpression of primate and human-specific lncRNAs are sources of molecular evolution and dysfunction of cortical excitatory neurons.
Sequence-based machine learning reveals 3D genome differences between bonobos and chimpanzees
Authors: Brand Colin M. , Kuang Shuzhen , Gilbertson Erin N. , McArthur Evonne , Pollard Katherine S. , Webster Timothy H. , Capra John A. 
Published: October 2023

AbstractPhenotypic divergence between closely related species, including bonobos and chimpanzees (genusPan), is largely driven by variation in gene regulation. The 3D structure of the genome mediates gene expression; however, genome folding differences inPanare not well understood. Here, we apply machine learning to predict genome-wide 3D genome contact maps from DNA sequence for 56 bonobos and chimpanzees, encompassing all five extant lineages. We use a pairwise approach to estimate 3D divergence between individuals from the resulting contact maps in 4,420 1 Mb genomic windows. While most pairs were similar,∼17% were predicted to be substantially divergent in genome folding. The most dissimilar maps were largely driven by single individuals with rare variants that pro-duce unique 3D genome folding in a region. We also identified 89 genomic windows where bonobo and chimpanzee contact maps substantially diverged, including several windows harboring genes associated with traits implicated inPanphenotypic divergence. We usedin silicomutagenesis to identify 51 3D-modifying variants in these bonobo-chimpanzee diver-gent windows, finding that 34 or 66.67% induce genome folding changes via CTCF binding motif disruption. Our results reveal 3D genome variation at the population-level and identify genomic regions where changes in 3D folding may contribute to phenotypic differences in our closest living relatives.
How osteons form: A quantitative hypothesis-testing analysis of cortical pore filling and wall asymmetry
Authors: Hegarty-Cremer Solene G. D. , Borggaard Xenia G. , Andreasen Christina M. , van der Eerden Bram C. J. , Simpson Matthew J. , Andersen Thomas L. , Buenzli Pascal R. 
Published: October 2023

AbstractOsteon morphology provides valuable information about the interplay between different processes involved in bone remodelling. The correct quantitative interpretation of these morphological features is challenging due to the complexity of interactions between osteoblast behaviour, and the evolving geometry of cortical pores during pore closing. We present a combined experimental and mathematical modelling study to provide insights into bone formation mechanisms during cortical bone remodelling based on histological cross-sections of quiescent human osteons and hypothesis-testing analyses. We introduce wall thickness asymmetry as a measure of the local asymmetry of bone formation within an osteon and examine the frequency distribution of wall thickness asymmetry in cortical osteons from human iliac crest bone samples from women 16–78 years old. Our measurements show that most osteons possess some degree of asymmetry, and that the average degree of osteon asymmetry in cortical bone evolves with age. We then propose a comprehensive mathematical model of cortical pore filling that includes osteoblast secretory activity, osteoblast elimination, osteoblast embedment as osteocytes, and osteoblast crowding and redistribution along the bone surface. The mathematical model is first calibrated to symmetric osteon data, and then used to test three mechanisms of asymmetric wall formation against osteon data: (i) delays in the onset of infilling around the cement line; (ii) heterogeneous osteoblastogenesis around the bone perimeter; and (iii) heterogeneous osteoblast secretory rate around the bone perimeter. Our results suggest that wall thickness asymmetry due to off-centred Haversian pores within osteons, and that nonuniform lamellar thicknesses within osteons are important morphological features that can indicate the prevalence of specific asymmetry-generating mechanisms. This has significant implications for the study of disruptions of bone formation as it could indicate what biological bone formation processes may become disrupted with age or disease.
Overcoming antibody-resistant SARS-CoV-2 variants with bispecific antibodies constructed using non-neutralizing antibodies
Authors: Inoue Tetsuya , Yamamoto Yuichiro , Sato Kaoru , Nakamura Yuko , Shimizu Yoshimi , Ogawa Motohiko , Onodera Taishi , Takahashi Yoshimasa , Wakita Takaji , Kaneko Mika K. , Fukasawa Masayoshi , Kato Yukinari , Noguchi Kohji 
Published: October 2023

SUMMARYA current challenge is the emergence of SARS-CoV-2 variants, such as BQ.1.1 and XBB.1.5, that can evade immune defenses, thereby limiting antibody drug effectiveness. Emergency-use antibody drugs, including the widely effective bebtelovimab, are losing their benefits. One potential approach to address this issue are bispecific antibodies which combine the targeting abilities of two antibodies with distinct epitopes. We engineered neutralizing bispecific antibodies in the IgG-scFv format from two initially non-neutralizing antibodies, CvMab-6 (which binds to the receptor-binding domain [RBD]) and CvMab-62 (targeting a spike protein S2 subunit epitope adjacent to the known anti-S2 antibody epitope). Furthermore, we created a bispecific antibody by incorporating the scFv of bebtelovimab with our anti-S2 antibody, demonstrating significant restoration of effectiveness against bebtelovimab-resistant BQ.1.1 variants. This study highlights the potential of neutralizing bispecific antibodies, which combine existing less effective anti-RBD antibodies with anti-S2 antibodies, to revive the effectiveness of antibody therapeutics compromised by immune-evading variants.