Biology subjects

Cavallo, F.

Publications and source records attributed to Cavallo, F..

4 recordsLinked to original sources

bioRxiv2025

A flexible end-to-end automated sample preparation workflow enables reproducible large-scale bottom-up proteomics

Bottom-up proteomics holds significant promise for clinical applications due to its high sensitivity and precision, but is limited by labor-intensive, low-throughput sample preparation methods. Advanced automation is essential to enhance throughput, reproducibility, and accuracy and to allow standardization to make bottom-up proteomics amenable for large-scale studies. We developed a fully integrated, automated sample preparation platform that covers the entire process from biological sample input to mass spectrometry-ready peptide output and can be applied on a multitude of biological samples. With this end-to-end solution, we achieved high intra- and inter-plate reproducibility, as well as longitudinal consistency, resulting in precise and reproducible workflows. We showed that our automated workflow surpasses established manual and semi-automated workflows, while improving time efficiency. Finally, we demonstrated the suitability of our automated sample preparation platform for drug development by performing a high-content compound characterization for targeted protein degradation, where high throughput and quantitative accuracy are indispensable. For this, we coupled application-specific workflows to perform proteome profiling and confirm target degradation by precise protein quantification. Overall, our results highlight the selective degradation of specific proteins of interest for ten selected compounds across two cell lines. Thus, the automated sample preparation platform facilitates rapid adaptation to emerging developments in proteomics sample preparation, combining standardization, flexibility, and high-throughput capabilities to drive significant advancements in clinical assays and proteomics research.

biochemistry↗

bioRxiv2024

Lead in the head: persistent lead poisoning of waterfowl in the Camargue (southern France) ten years after the ban on the use of lead shot in wetlands

Lead pollution remains a worldwide environmental and health issue with persistent detrimental effects on human and wildlife. Despite having been identified as an issue for wildlife a long time ago, the use of lead shot for hunting in wetlands was banned only in 2006 in France. Here, we took advantage of a long-term monitoring of waterfowl lead shot contamination in the Camargue (southern France) to (1) assess the local effectiveness of the French regulation in reducing waterfowl contamination and to (2) assess local compliance regarding the use of nontoxic shots in wetlands from 2007 onwards. We collected waterfowl gizzard from 38 hunters in the Camargue during 20 hunting seasons (1998 to 2017) for a total of 2306 gizzards from 28 different species. From 2008 to 2019, we also systematically collected the cartridge casings at three sites of communal wetland hunting. Ratio of lead vs nontoxic cartridges found in the field allowed monitoring hunter compliance with the new regulation. Despite a ten-year long ban on lead shot our results do not show any significant reduction in waterfowl exposure to lead through shot ingestion in the Camargue over the 20-year monitoring period. Indeed, gizzards of harvested waterfowl were as likely to contain at least one lead shot before the ban as after with a mean prevalence of 12% over the 13 species considered across the study period. Cartridge casings showed the persistence of use of lead shots by hunters in the Camargue wetlands despite a slow increase in the use of nontoxic cartridges. This unequivocally indicates that the law is far from being strictly applied, pointing to an insufficient policy enforcement. Our results support the need for a complete ban of lead ammunition for both wetland and terrestrial wildfowl which will facilitate policy enforcement and compliance.

ecology↗

bioRxiv2021

The Amot/Integrin protein complex transmits mechanical forces required for vascular expansion

Vascular development is a complex multistep process involving the coordination of cellular functions such as migration, proliferation and differentiation. Understanding the underlying mechanisms of these processes is of importance due to involvement of vessel expansion in various pathologies. How mechanical forces generated by cells and transmission of these physical forces control vascular development is poorly understood. Using an endothelial-specific genetic model in mice, we show that deletion of the scaffold protein, Angiomotin (Amot), inhibits migration and expansion of physical and pathological vascular network. We further show that Amot is required for tip cell migration and the extension of cellular filopodia. Exploiting in vivo and in vitro molecular approaches, we show that Amot binds talin and is essential for relaying forces between fibronectin and the cytoskeleton. Finally, we provide evidence that Amot is a novel component of the endothelial integrin adhesome and propose that Amot integrates spatial cues from the extra-cellular matrix in order to form a functional vascular network.

cell biology↗

bioRxiv2017

From Cohorts to Molecules: Adverse Impacts of Endocrine Disrupting Mixtures

Convergent evidence associates endocrine disrupting chemicals (EDCs) with major, increasingly-prevalent human disorders. Regulation requires elucidation of EDC-triggered molecular events causally linked to adverse health outcomes, but two factors limit their identification. First, experiments frequently use individual chemicals, whereas real life entails simultaneous exposure to multiple EDCs. Second, population-based and experimental studies are seldom integrated. This drawback was exacerbated until recently by lack of physiopathologically meaningful human experimental systems that link epidemiological data with results from model organisms.\n\nWe developed a novel approach, integrating epidemiological with experimental evidence. Starting from 1,874 mother-child pairs we identified mixtures of chemicals, measured during early pregnancy, associated with language delay or low-birth weight in offspring. These mixtures were then tested on multiple complementary in vitro and in vivo models. We demonstrate that each EDC mixture, at levels found in pregnant women, disrupts hormone-regulated and disease-relevant gene regulatory networks at both the cellular and organismal scale.

molecular biology↗