Project description:Decreased mineral density is a risk factor for skeletal pathologies including bone metastasis, the leading cause of mortality in patients with advanced breast cancer, but the underlying mechanisms are poorly understood. While reduced mineral density can drive pathological bone remodeling via direct effects on select cell types, indirect effects due to broad changes of the microenvironment may be similarly important. However, how bone mineral content affects microenvironmental heterogeneity remains to be elucidated. Here, we leverage decellularized bone matrices with varied mineral content in combination with single-cell RNA-sequencing to study how reduced bone mineral content affects microenvironmental complexity and tumor growth. We performed single-cell RNA sequencing on implanted decellularized bovine bone scaffolds in which the mineral was either maintained at physiological levels or removed to simulate scenarios of impaired bone mineralization as, for example, present during aging. Using this approach, we explored the heterogeneous stromal response to varied bone mineral content in both an immunocompromised and immunocompetent, syngeneic mouse model in the presence and absence of cancer cells.
Project description:Gene expression profile analysis allowed to identify a panel of genes characteristic of silica materials effect on transformation process.
Project description:Despite considerable efforts to characterize the microbial ecology of the built environment, the metabolic mechanisms underpinning microbial colonization and successional dynamics remain unclear, particularly at high moisture conditions. Here, we applied bacterial/viral particle counting, qPCR, amplicon sequencing of the genes encoding 16S and ITS rRNA, and metabolomics to longitudinally characterize the ecological dynamics of four common building materials maintained at high humidity. We varied the natural inoculum provided to each material and wet half of the samples to simulate a potable water leak. Wetted materials had higher growth rates and lower alpha diversity compared to non-wetted materials, and wetting described the majority of the variance in bacterial, fungal, and metabolite structure. Inoculation location was weakly associated with bacterial and fungal beta diversity. Material type influenced bacterial and viral particle abundance and bacterial and metabolic (but not fungal) diversity. Metabolites indicative of microbial activity were identified, and they too differed by material.
Project description:Spiders are renowned for their efficient capture of flying insects using intricate aerial webs. How the spider nervous systems evolved to cope with this specialized hunting strategy and various environmental clues in an aerial space remains unknown. Here, we report a brain cell atlas of >30,000 single-cell transcriptomes from a web-building spider (Hylyphantes graminicola). Our analysis revealed the preservation of ancestral neuron types in spiders, including the potential coexistence of noradrenergic and octopaminergic neurons, and many peptidergic neuronal types that are lost in insects. By comparing the genome of two newly sequenced plesiomorphic burrowing spiders with three aerial web-building spiders, we found that the positively selected genes in the ancestral branch of web-building spiders were preferentially expressed (42%) in the brain, especially in the three mushroom body-like neuronal types. By gene enrichment analysis and RNAi experiments, these genes were suggested to be involved in the learning and memory pathway and may influence the spiders’ web-building and hunting behavior. Our results provide key sources for understanding the evolution of behavior in spiders and reveal how molecular evolution drives neuron innovation and the diversification of associated complex behaviors.
Project description:Concerning the roles of LCN2 in chronic inflammation and fibrosis, we investigated chronic liver inflammation and fibrosis using repeated carbon tetrachloride (CCl4) in mineral oil injection. We found that mice treated with the mineral oil vehicle alone also showed liver inflammation and more severe in wild-type mice compared to lipocalin 2 null mice.
Project description:Cell lysates of Fusarium sp. DS 682, a saprotrophic fungus, were prepared from fungal biomass grown on PDA agar without M9 and micronutrients. Fungus was exposed to minerals, 100 uL of 50% (w/v) natural kaolinite solution. Three fungal plate collections for each growth condition, with (+ mineral) and without mineral (- mineral), were grown for 14 days on PDA (incubated at 28 C) and harvested after 15 days. Extracted hyphae (500 mg) was prepared using the MPLEx protocol. A nanoACQUITY ultra performance liquid chromatography (LC) with a 2DLC system was used for separation of protein digests. Eluted peptides from the C18 column were analyzed using a Q-Exactive Plus Orbitrap MS for high resolution MS and high-energy collision-induced dissociation tandem MS by electrospray ionization for subsequent quantitative proteomic analysis. Data was searched with MaxQuant. Additional post-processed results and metadata can be accessed at https://doi.org/10.25584/KSOmicsFspDS682/1766303