Antibodies are known to have an important role in the development of rheumatoid arthritis (RA), one of the most prevalent chronic inflammatory diseases which primarily involves the joints. Most RA patients develop autoantibodies against immunoglobulin G (IgG) and changes in IgG glycosylation have been associated with RA. We undertook this study to determine whether altered IgG glycosylation precedes the disease diagnosis. We studied IgG glycosylation in RA in two prospective cohorts (N = 14,749) by measuring 28 IgG glycan traits in 179 subjects who developed RA within 10-years follow-up and 358 matched controls. Ultra-performance liquid chromatography method based on hydrophilic interactions (HILIC-UPLC) was used to analyse IgG glycans. Future RA diagnosis associated with traits related to lower galactosylation and sialylation of IgG when comparing the cases to the matched controls. In RA cases, these traits did not correlate with the time between being recruited to the study and being diagnosed with RA (median time 4.31 years). The difference in IgG glycosylation was relatively stable and present years before diagnosis. This indicates that long-acting factors affecting IgG glycome composition are among the underlying mechanisms of RA and that decreased galactosylation is a pre-existing risk factor involved in the disease development.
Polyphenols are secondary plant metabolites that possess potentially health-promoting properties and which occur in various edible plants and plant products. Especially the stilbenoid resveratrol has been extensively studied regarding its anticarcinogenic and chemopreventive activities. However, research has recently focused on the investigation of other natural or synthetic compounds in order to find substances that show a higher bioactivity and/or bioavailability than resveratrol. In this context, we exemplarily investigated the cytotoxic/growth-inhibiting properties of the resveratrol tetramer r-viniferin on the prostate cancer cell line LNCaP and compared them with those of resveratrol. By using the sulforhodamine B assay followed by cell cycle analysis via flow cytometry and commercially available apoptosis/necrosis assay kits, we show that both compounds were able to inhibit the growth of LNCaP cells and to induce a cell cycle arrest in the G1 phase. However, r-viniferin was significantly more potent in inhibiting cellular growth than resveratrol and the only compound that increased the apoptotic cellular fraction as well as the activity of apoptosis-associated enzymes. In conclusion, r-viniferin leads to cytotoxicity in LNCaP cells at fairly low concentrations, and it is therefore conceivable that it might be used as a chemopreventive agent or as an adjuvant in prostate cancer therapy.
Fibrinogen (FIB) is a secretory glycoprotein synthesized by hepatocytes that has a key role in blood clotting. Its glycosylation has not been studied in detail and little is known about the biological variability of FIB N-glycosylation, mainly due to the lack of fast, simple, and robust approaches to purify FIB from blood plasma samples. In recent years, customised chromatographic monoliths have been used for a variety of biological applications due to their unique characteristics. Here we describe development and optimisation of monolithic supports bearing monoclonal anti-human fibrinogen antibodies in a single column as well as in multi-well plate formats with high FIB specificity and binding capacity for fast immunoaffinity purification of FIB from human blood samples. The developed semi-high-throughput workflow has been successfully applied for FIB immunoaffinity isolation and subsequent ultra performance liquid chromatography N-glycosylation analysis in ten healthy human individuals, demonstrating the potential of monolithic supports in glycomics studies.
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