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Universal and Ultrasensitive Immunochromatographic Assay by Using an Antigen as a Bifunctional Element and Antialbumin Antibody on a Test Line.

A universal and ultrasensitive immunochromatographic assay (ICA) was established using antigen as a bifunctional element and antialbumin antibody in a test line. Preincubation was introduced for competitive recognition. After optimization, the linear detection of aflatoxin M (AFM) with quantum dot bead (QB)-based ICA (QB-ICA) sensor ranged from 10 to 52 pg mL, with a 50% inhibitory concentration (IC) of 23 pg mL, which was nearly 49.6-fold lower than those of ICA on a traditional structure with traditional pretreatment (IC = 1.10 ng mL) and 10-fold lower than those of ICA on a traditional structure with acid aid pretreatment (IC = 0.25 ng mL). The limit of detection (LOD) for AFM was 16 pg mL in milk, which was approximately 16.3-fold times higher than those of ICA on a traditional structure with traditional pretreatment and 6.3-fold higher than those of ICA on a traditional structure with acid aid pretreatment. The LOD improved by 20-fold by using the proposed structure compared to that of conventional enzyme-linked immunosorbent assay (ELISA) for AFM-spiked milk samples (IC = 0.12 ng mL). The performance and practicability of the established QB-ICA sensor were validated with a commercial ELISA kit. To evaluate universality, we successfully detected chloramphenicol, with IC of 0.42 ng mL. Given its high sensitivity and universality, the proposed QB-ICA can be used as an alternative for rapid, sensitive, and universal quantitative detection of all small-molecule analytes.

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Immune Dysfunction and Albumin-Related Immunity in Liver Cirrhosis.

Liver cirrhosis yearly causes 1.2 million deaths worldwide, ranking as the 10th leading cause of death in the most developed countries. High susceptibility to infections along with a significant risk for infection-related mortality justifies the description of liver cirrhosis as the world's most common immunodeficiency syndrome. Liver cirrhosis is an end-stage organic disease hallmarked by a multifaceted immune dysfunction due to deterioration of antimicrobial recognition and elimination mechanisms in macrophages along with an impaired antigen presentation ability in circulating monocytes. Bacterial translocation supports-and is supported by-uncontrolled activation of immune cell responses and/or loss of toll-like receptor (TLR) tolerance, which can turn exaggerated inflammatory responses to systemic inflammation. Lipopolysaccharide (LPS) or endotoxin boosts systemic inflammatory activity through activation of TLR-2- and TLR-4-dependent pathways and facilitate a massive production of cytokines. This, in turn, results into elevated secretion of reactive oxygen species (ROS), which further enhances intestinal hyperpermeability and thus sustains a vicious circle of events widely known as "leaky gut." Albumin can be of particular benefit in cirrhotic patients with spontaneous bacterial peritonitis and/or hepatorenal syndrome type of acute kidney injury (HRS-AKI) due to anti-inflammatory and antioxidative stress as well as volume-expanding properties and endothelial-stabilizing attributes. However, presence of autoantibodies against albumin in patients with liver cirrhosis has been described. Although previous research suggested that these antibodies should be regarded as naturally occurring antibodies (NOA), the origin of the antialbumin immune response is obscure. High occurrence of NAO/albumin complexes in patients with liver disease might reflect a limited clearance capacity due to bypassing portal circulation. Moreover, high burden of oxidized albumin is associated with less favorable outcome in patients with liver cirrhosis. To date, there is no data available as to whether oxidized forms of albumin result in neoepitopes recognized by the immune system. Nevertheless, it is reasonable to hypothesize that these alterations may have the potential to induce antialbumin immune responses and thus favor systemic inflammation.

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