Neutrophil-Mediated Experimental Metastasis Is Enhanced by VEGFR Inhibition in a Zebrafish Xenograft Model

Inhibition of VEGF signalling effectively suppresses localized tumour growth but accelerates tumour invasiveness and micrometastasis by unknown mechanisms. To study the dynamic and reciprocal interactions between tumour cells and their microenvironment during these processes, we established a xenograft model by injecting tumour cells into the blood circulation of transparent zebrafish embryos. This reproducibly results in rapid simultaneous formation of a localized tumour and experimental micrometastasis, allowing time-resolved imaging of both processes at single-cell resolution within 1 week. The tumour vasculature was initiated de novo by remodelling of primitive endothelial cells into a functional network. Roles of myeloid cells in critical tumourigenesis steps such as vascularization and invasion were revealed by genetic and pharmaceutical approaches. We discovered that the physiological migration of neutrophils controlled tumour invasion by conditioning the collagen matrix and forming the metastatic niche, as detected by two-photon confocal microscopy and second harmonic generation. Administration of VEGFR inhibitors blocked tumour vascularization and a localized tumour growth but enhanced migration of neutrophils, which in turn promoted tumour invasion and formation of micrometastasis. This demonstrates the in vivo cooperation between VEGF signalling and myeloid cells in metastasis and provides a new mechanism underlying the recent findings that VEGFR targeting can promote tumour invasiveness

ERK1 and ERK2 MAPK are key regulators of distinct gene sets in zebrafish embryogenesis

<p>Abstract</p> <p>Background</p> <p>The MAPK signaling proteins are involved in many eukaryotic cellular processes and signaling networks. However, specific functions of most of these proteins in vertebrate development remain elusive because of potential redundancies. For instance, the upstream activation pathways for ERK1 and ERK2 are highly similar, and also many of their known downstream targets are common. In contrast, mice and zebrafish studies indicate distinct roles for both ERKs in cellular proliferation, oncogenic transformation and development. A major bottleneck for further studies is that relatively little is known of i<it>n vivo </it>downstream signaling specific for these kinases.</p> <p>Results</p> <p>Microarray based gene expression profiling of ERK1 and ERK2 knockdown zebrafish embryos at various stages of early embryogenesis resulted in specific gene expression signature sets that showed pronounced differences in gene ontology analyses. In order to predict functions of these genes, zebrafish specific <it>in silico </it>signaling pathways involved in early embryogenesis were constructed using the GenMAPP program. The obtained transcriptome signatures were analyzed in the BMP, FGF, Nodal and Wnt pathways. Predicted downstream effects of ERK1 and ERK2 knockdown treatments on key pathways responsible for mesendoderm development were confirmed by whole mount in situ hybridization experiments.</p> <p>Conclusion</p> <p>The gene ontology analyses showed that ERK1 and ERK2 target common and distinct gene sets, confirming the difference in knockdown phenotypes and diverse roles for these kinases during embryogenesis. For ERK1 we identified specific genes involved in dorsal-ventral patterning and subsequent embryonic cell migration. For ERK2 we identified genes involved in cell-migration, mesendoderm differentiation and patterning.</p> <p>The specific function of ERK2 in the initiation, maintenance and patterning of mesoderm and endoderm formation was biologically confirmed.</p

A survey on fairness of large language models in e-commerce: progress, application, and challenge

This survey explores the fairness of large language models (LLMs) in e-commerce, examining their progress, applications, and the challenges they face. LLMs have become pivotal in the e-commerce domain, offering innovative solutions and enhancing customer experiences. This work presents a comprehensive survey on the applications and challenges of LLMs in e-commerce. The paper begins by introducing the key principles underlying the use of LLMs in e-commerce, detailing the processes of pretraining, fine-tuning, and prompting that tailor these models to specific needs. It then explores the varied applications of LLMs in e-commerce, including product reviews, where they synthesize and analyze customer feedback; product recommendations, where they leverage consumer data to suggest relevant items; product information translation, enhancing global accessibility; and product question and answer sections, where they automate customer support. The paper critically addresses the fairness challenges in e-commerce, highlighting how biases in training data and algorithms can lead to unfair outcomes, such as reinforcing stereotypes or discriminating against certain groups. These issues not only undermine consumer trust, but also raise ethical and legal concerns. Finally, the work outlines future research directions, emphasizing the need for more equitable and transparent LLMs in e-commerce. It advocates for ongoing efforts to mitigate biases and improve the fairness of these systems, ensuring they serve diverse global markets effectively and ethically. Through this comprehensive analysis, the survey provides a holistic view of the current landscape of LLMs in e-commerce, offering insights into their potential and limitations, and guiding future endeavors in creating fairer and more inclusive e-commerce environments.Comment: 21 pages, 9 figure

Identification of novel potential hypoxia-inducible factor-1α inhibitors through machine learning and computational simulations

IntroductionHypoxia-inducible factor-1α (HIF-1α) has become a significant therapeutic target for breast cancer and other cancers by regulating the expression of downstream genes such as erythropoietin, thereby improving cell survival in hypoxic conditions.MethodsWe jointly applied a multistage screening system encompassing machine learning, molecular docking, and molecular dynamics simulations to conduct virtual screening of the “Traditional Chinese Medicine Monomer Library” for potential HIF-1α inhibitors. The virtual screening was conducted in three sequential stages, applying the following selection criteria sequentially: an activity prediction score greater than or equal to 0.8, a stronger binding affinity, and an MM-PBSA binding free energy lower than the reference compound.Results and DiscussionWe retrieved 361 compounds with HIF-1α inhibitory activity data from the ChEMBL database for the construction and evaluation of machine learning models. Among the six constructed models, the random forest model based on RDKit molecular descriptor with the optimal comprehensive performance was employed for virtual screening. Ultimately, four compounds were selected for binding mode analyses and 100 ns molecular dynamics simulations. The results showed that the compounds Arnidiol and Epifriedelanol exhibit the most stable interactions with the HIF-1α protein, which can serve as potential HIF-1α inhibitors for future investigations

Impact of Lactobacillus casei BL23 on the Host Transcriptome, Growth and Disease Resistance in Larval Zebrafish

In this study, zebrafish were treated with Lactobacillus strains as probiotics from hatching to puberty, and the effect of treatment with L. casei BL23 on the development and immunity response of the host was investigated. Genes that were differentially expressed (DEGs) in the overall body and intestine were detected at 14 days post fertilization (dpf) and 35 dpf, respectively, using whole transcriptome sequencing (mRNAseq). We showed that zebrafish raised by continuous immersion with L. casei BL23 showed a higher final body weight at 14 dpf (P &lt; 0.05), and 35 dpf (P &lt; 0.01). DEGs between L. casei BL23 treatment and control group at 14 dpf were involved in myogenesis, cell adhesion, transcription regulation and DNA-binding and activator. At 35 dpf, 369 genes were DEGs in the intestine after treatment with L. casei BL23, which were involved in such categories as signaling, secretion, motor proteins, oxidoreductase and iron, tight junctions, lipid metabolism, growth regulation, proteases, and humoral and cellular effectors. KEGG analysis showed DEGs to be involved in such pathways as those associated with tight junctions and the PPAR signal pathway. RT-qPCR analysis showed that expression of insulin-like growth factors-I (igf1), peroxisome proliferator activated receptors-α (ppar-α) and -β (ppar-β), Vitamin D receptor-α (vdr-α), and retinoic acid receptor-γ (rar-γ) was up-regulated in fish treated with L. casei BL23 at 35 dpf. After 35 days of treatment, the mortality rate in L. casei BL23 treated group was lower than the control after challenge with A. hydrophila (P &lt; 0.05), and the pro-inflammatory cytokine il-1β, anti-inflammatory cytokine il-10 and complement component 3a (c3a) showed more expression in L. casei BL23 group at 8h after challenge, 24 h after challenge, or both.. Together, these data suggest that specific Lactobacillus probiotic strains can accelerate the development profile and enhance immunity in zebrafish, which supports the rationale of early administration of probiotics in aquaculture

Automated Whole Animal Bio-Imaging Assay for Human Cancer Dissemination

A quantitative bio-imaging platform is developed for analysis of human cancer dissemination in a short-term vertebrate xenotransplantation assay. Six days after implantation of cancer cells in zebrafish embryos, automated imaging in 96 well plates coupled to image analysis algorithms quantifies spreading throughout the host. Findings in this model correlate with behavior in long-term rodent xenograft models for panels of poorly- versus highly malignant cell lines derived from breast, colorectal, and prostate cancer. In addition, cancer cells with scattered mesenchymal characteristics show higher dissemination capacity than cell types with epithelial appearance. Moreover, RNA interference establishes the metastasis-suppressor role for E-cadherin in this model. This automated quantitative whole animal bio-imaging assay can serve as a first-line in vivo screening step in the anti-cancer drug target discovery pipeline

Essential tremor-challenged maxillary rehabilitation using a digitally guided all-on-six implant restoration: a case report

This case report describes a 73-year-old female with essential tremor who experienced significant dissatisfaction with her maxillary complete denture. Her tremor-related dexterity impairment limited her ability to manage removable prostheses, while the excessive denture volume severely disrupted speech. Moreover, her neuromuscular condition made it challenging to adapt to a new occlusal scheme. To address these issues, a digitally guided all-on-six implant-supported prosthesis was delivered under local anesthesia, preserving her pre-existing, neuromuscularly adapted occlusion with minor esthetic adjustments to the anterior teeth. At the 1-year follow-up, the patient reported high satisfaction with both esthetics and function, along with improved speech and masticatory efficiency. This report underscores the value of digital workflows in maintaining functional adaptation and providing predictable prosthodontic solutions for patients with neuromuscular impairments

Genome-wide identification and expression analysis of sucrose phosphate synthase and sucrose-6-phosphate phosphatase family genes in Camellia sinensis

Sucrose phosphate synthetase (SPS, EC 2.4.1.14) and sucrose phosphate phosphatase (SPP, EC 3.1.3.24) are two key enzymes for sucrose biosynthesis, which play essential roles in growth, development and stress responses of plants. However, the roles of SPS and SPP have not been illustrated well in tea plants until now. In this study, a unique CsSPP and five CsSPSs (CsSPS1-5) were identified from the tea plant genome. Bioinformatic analysis results found that CsSPP and CsSPSs were clustered together with the known SPPs and SPSs of other species, respectively, and their amino acid sequences contain the conserved domains required for sucrose biosynthesis. Tissue-specific analysis results showed that CsSPP and CsSPSs were widely involved in vegetative and reproductive growth of tea plant, among which the transcription levels of CsSPP was highest in immature stem, while CsSPSs were highest in flower. Spatio-temporal expression analysis results showed that all of these genes are involved in abiotic stress responses of tea plants. Meanwhile, SPS activity and the contents of sucrose, glucose, fructose, and total soluble sugar in 'Shuchazao' cultivar increased more than that of 'Baiye1' under low temperature conditions. Correlation analysis results showed that the expression profiles of CsSPS2/4/5 were significantly and positively correlated with sucrose content in 'Shuchazao' cultivars under low temperature conditions, suggesting the significant roles of these genes in sucrose accumulation. In conclusion, this study will provide a theoretical basis for further functional research of SPS and SPP in plants