Single-molecule force spectroscopy (SMFS) uses the cantilever tip of an AFM to apply a force able to unfold a single protein. The obtained force-distance curve encodes the unfolding pathway, and from its analysis it is possible to characterize the folded domains. SMFS has been mostly used to study the unfolding of purified proteins, in solution or reconstituted in a lipid bilayer. Here, we describe a pipeline for analyzing membrane proteins based on SMFS, that involves the isolation of the plasma membrane of single cells and the harvesting of force-distance curves directly from it. We characterized and identified the embedded membrane proteins combining, within a Bayesian framework, the information of the shape of the obtained curves, with the information from Mass Spectrometry and proteomic databases. The pipeline was tested with purified/reconstituted proteins and applied to five cell types where we classified the unfolding of their most abundant membrane proteins. We validated our pipeline by overexpressing 4 constructs, and this allowed us to gather structural insights of the identified proteins, revealing variable elements in the loop regions. Our results set the basis for the investigation of the unfolding of membrane proteins in situ, and for performing proteomics from a membrane fragment.

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Although prototypical network (ProtoNet) has proved to be an effective method for few-shot sound event detection, two problems still exist. Firstly, the small-scaled support set is insufficient so that the class prototypes may not represent the class center accurately. Secondly, the feature extractor is taskagnostic (or class-agnostic): the feature extractor is trained with base-class data and directly applied to unseen-class data.To address these issues, we present a novel mutual learning framework with transductive learning, which aims at iteratively updating the class prototypes and feature extractor. More specifically, we propose to update class prototypes with transductive inference to make the class prototypes as close to the true class center as possible. To make the feature extractor to be task-specific, we propose to use the updated class prototypes to fine-tune the feature extractor. After that, a finetuned feature extractor further helps produce better class prototypes. Our method achieves the F-score of 38.4% on the DCASE 2021 Task 5 evaluation set, which won the first place in the few-shot bioacoustic event detection task of Detection and Classification of Acoustic Scenes and Events (DCASE) 2021 Challenge.