1,721,071 research outputs found
Advanced correlative light/electron microscopy: current methods and new developments using Tokuyasu cryosections
No full textMicroscopy is an essential tool for analysis of cellular structures and function. With the advent of new fluorescent probes and super-resolution light microscopy techniques, the study of dynamic processes in living cells has been greatly facilitated. Fluorescence light microscopy provides analytical, quantitative, and three-dimensional (3D) data with emphasis on analysis of live cells using fluorescent markers. Sample preparation is easy and relatively inexpensive, and the use of appropriate tags provides the ability to track specific proteins of interest. Of course, only electron microscopy (EM) achieves the highest definition in terms of ultrastructure and protein labeling. To fill the gap between light microscopy and EM, correlative light and electron microscopy (CLEM) strategies have been developed. In particular, hybrid techniques based upon immuno-EM provide sensitive protein detection combined with high-resolution information on cell structures and protein localization. By adding the third dimension to EM with electron tomography (ET) combined with rapid freezing, CLEM techniques now provide additional tools for quantitative 3D analysis. Here, we overview the major methods applied and highlight the latest advances in the field of CLEM. We then focus on two selected techniques that use cryosections as substrate for combined biomolecular imaging. Finally, we provide a perspective of future developments in the field
Cartilage in ageing and osteoarthritis: altered expression of fibronectin and related integrin receptors
No full textCartilage in ageing and osteoarthritis: altered expression of fibronectin and related integrin receptors.
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Cartilage in ageing and osteoarthritis: altered expression of fibronectin and related integrin receptors
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Moving Toward Implementation of Responsible Artificial Intelligence in Health Care: The European TRAIN Initiative
No full textThe S-RACE Platform: A cloud-based solution for real-world health data, driving clinical translation and responsible governance of artificial intelligence
No full textThe S-RACE platform is a cloud-based AI solution for using real-world health data. It addresses data quality and governance challenges with an end-to-end pipeline, including on-premise anonymisation and tools for clinicians and data scientists. Aligned with responsible AI principles, it aims to accelerate the translation of AI research into clinical practice, improving patient care
Use of human chondrocyte cell cultures to identify and characterize reactive antibodies in rheumatoid arthritis sera
No full textOBJECTIVE:
To study the reactivity of rheumatoid arthritis (RA) sera with human chondrocyte populations isolated from normal cartilage and expanded in vitro.
METHODS:
Human articular chondrocytes were cultured as adherent (non-differentiated) cells on plastic dishes or in suspension (differentiated) on dishes previously coated with a thin layer of 1% agarose. Sera from 28 RA patients and 5 paired synovial fluids were tested on lysates from chondrocytes and fibroblasts as control by immunoblot. Antigen expression on the cell membrane was evaluated by flow cytometry in a few sera.
RESULTS:
In 9/28 RA sera IgG antibodies specific for chondrocyte antigens (97 kDa, 74 kDa, 67 kDa, 60 kDa, 54 kDa, 48 kDa and 37 kDa) were detected. Twelve sera reacted with proteins expressed both on chondrocytes and fibroblasts and 7 with fibroblasts only; two sera had no reactivity. When lysates from adherent or suspension chondrocytes were compared, RA sera reacted with higher intensity and detected more antigens on chondrocytes cultured in suspension. Flow cytometry assay demonstrated that RA sera are able to recognize antigens expressed on the cell membrane of the human chondrocytes.
CONCLUSION:
Our data indicate that: a) 32% of the RA sera contain antibodies reactive with antigens expressed exclusively by chondrocytes, but this value rises to 75% if antigens expressed both by chondrocytes and fibroblasts are considered; b) the reactivity of fully differentiated chondrocytes in suspension culture is higher than the reactivity of chondrocytes cultured in monolayer; and c) some of the chondrocyte-specific antigens identified are associated with the chondrocyte membrane. Thus, in vitro cultured chondrocytes may be used to study both the specificity and the biological activity of autoantibodies in RA
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