Endothelial dysfunction characterized by decreased nitric oxide (NO) bioavailability is the first stage of coronary artery disease. It is known that one of the factors associated with an increased risk of coronary artery disease is a high plasma level of uric acid. However, causative associations between hyperuricaemia and cardiovascular risk have not been definitely proved. In this work, we tested the effect of uric acid on endothelial NO bioavailability. Electrochemical measurement of NO production in acetylcholine-stimulated human umbilical endothelial cells (HUVECs) revealed that uric acid markedly decreases NO release. This finding was confirmed by organ bath experiments on mouse aortic segments. Uric acid dose-dependently reduced endothelium-dependent vasorelaxation. To reveal the mechanism of decreasing NO bioavailability we tested the effect of uric acid on reactive oxygen species production by HUVECs, on arginase activity, and on acetylcholine-induced endothelial NO synthase phosphorylation. It was found that uric acid increases arginase activity and reduces endothelial NO synthase phosphorylation. Interestingly, uric acid significantly increased intracellular superoxide formation. In conclusion, uric acid decreases NO bioavailability by means of multiple mechanisms. This finding supports the idea of a causal association between hyperuricaemia and cardiovascular risk.

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It is known that cells and organisms can indirectly "sense" changes in L-arginine availability via changes in the activity of various metabolic pathways. However, the mechanism(s) by which genes can be directly regulated by L-arginine in mammalian cells have not yet been elucidated. We investigated the effect of L-arginine in the in vivo model of peritoneal inflammation in mice and in vitro in RAW 264.7 macrophages. A detailed analysis of basic physiological functions and selected intracellular signaling cascades revealed that L-arginine is crucial for the acceleration of macrophage activation by bacterial lipopolysaccharide. L-arginine increased the production of reactive oxygen species, nitric oxide, release of Ca(2+), as well as inducible nitric oxide synthase expression. Interestingly, the effect of L-arginine on macrophage activation was dependent on the phosphorylation of mitogen-activated protein kinases and activity of phospholipase C. In RAW 264.7 cells, L-arginine was shown to modulate the response of macrophages toward lipopolysaccharide via the activation of G-protein-coupled receptors. According to our data, we concluded that L-arginine availability plays a key role in the initiation of intracellular signaling pathways that trigger the lipopolysaccharide-induced inflammatory responses in murine macrophages. Although macrophages are partially stimulated in the absence of extracellular L-arginine, the presence of this amino acid significantly accelerates the sensitivity of macrophages to bacterial endotoxin.

Significant mortalities associated with emerging viral diseases are challenging the economy of common carp aquaculture. As such, there is an increased need to disentangle how infected fish cope with progressive disease pathology and lose the ability for homeostatic maintenance of key physiological parameters. A natural carp edema virus (CEV) infection outbreak at a carp fish farm provided an opportunity to examine diseased and healthy carp in the same storage pond, thereby contributing to our better understanding of CEV disease pathophysiology. The disease status of fish was determined using PCR-based virus identification combined with analysis of gill pathology. Compared with healthy control carp, the blood chemistry profile of CEV-infected fish revealed major disruptions in electrolyte and acid-base balance (i.e., hyponatraemia, hypochloraemia, hyperphosphatemia, elevated pH, base excess, and anion gap and decreased partial dissolved carbon dioxide). In addition, we recorded hyperproteinaemia, hyperalbuminaemia, hypotonic dehydration, endogenous hyperammonaemia, and decreased lactate along with increased creatinine, alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase. Red blood cell associated hematology variables were also elevated. The multivariate pattern of responses for blood chemistry variables (driven by sodium, pH, partial dissolved carbon dioxide, ammonia, and albumin in the principal component analysis) clearly discriminated between CEV-infected and control carp. To conclude, we show that CEV infection in carp exerts complex adverse effects and results in severe metabolic disturbance due to the impaired gill respiratory and excretory functioning.