Expression and functional role of adenosine receptors in regulating inflammatory responses in human synoviocytes.

BACKGROUND AND PURPOSE: Adenosine is an endogenous modulator, interacting with four G-protein coupled receptors (A(1), A(2A), A(2B) and A(3)) and acts as a potent inhibitor of inflammatory processes in several tissues. So far, the functional effects modulated by adenosine receptors on human synoviocytes have not been investigated in detail. We evaluated mRNA, the protein levels, the functional role of adenosine receptors and their pharmacological modulation in human synoviocytes. EXPERIMENTAL APPROACH: mRNA, Western blotting, saturation and competition binding experiments, cyclic AMP, p38 mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-kappaB activation, tumour necrosis factor alpha (TNF-alpha) and interleukin-8 (IL-8) release were assessed in human synoviocytes isolated from patients with osteoarthritis. KEY RESULTS: mRNA and protein for A(1), A(2A), A(2B) and A(3) adenosine receptors are expressed in human synoviocytes. Standard adenosine agonists and antagonists showed affinity values in the nanomolar range and were coupled to stimulation or inhibition of adenylyl cyclase. Activation of A(2A) and A(3) adenosine receptors inhibited p38 MAPK and NF-kappaB pathways, an effect abolished by selective adenosine antagonists. A(2A) and A(3) receptor agonists decreased TNF-alpha and IL-8 production. The phosphoinositide 3-kinase or G(s) pathways were involved in the functional responses of A(3) or A(2A) adenosine receptors. Synoviocyte A(1) and A(2B) adenosine receptors were not implicated in the inflammatory process whereas stimulation of A(2A) and A(3) adenosine receptors was closely associated with a down-regulation of the inflammatory status. CONCLUSIONS AND IMPLICATIONS: These results indicate that A(2A) and A(3) adenosine receptors may represent a potential target in therapeutic modulation of joint inflammation.

Enzyme-linked immunosorbent assay (ELISA) based on superparamagnetic nanoparticles for aflatoxin M1 detection.

Five different clones of antibodies developed against the aflatoxin M(1) were investigated by using the classical indirect and direct competitive Enzyme-Linked Immunosorbent Assay (ELISA) formats, and also the direct competitive ELISA based on the use of the superparamagnetic nanoparticles. The purpose of this study was to assess if not so friendly time classical ELISA procedures can be further improved, by reducing the coating, blocking and competition time. Here we showed that a complete dc-ELISA (coating, blocking and competition step) based on the use of superparamagnetic nanoparticles can be performed in basically 40 min, if coating step (20 min) should be taken into account. Moreover, the standard analytical characteristics of the proposed method fulfil the requirements for detecting AFM(1) in milk, in a wide linear working range (4-250 ng/L). The IC(50) value is 15 ng/L. The matrix effect and the recovery rate were assessed, using the European Reference Material (BD282, zero level of AFM(1)), showing an excellent percentage of recovery, close to 100%.

[To live death in the hospital. The Lendinara School of Nursing]. / Vivere la morte in ospedale. Scuola Infermieri di Lendinara.

Issues related to death and dying are important subjects in the educational curriculum of nursing students; in the nursing school of Lendinara this subjects were discussed analysing practical and emotional experience of nursing students. An anonymous questionnaire was distributed to the 74 second and third year students (67 returned) to explore emotional experience and 52 terminal patients were observed during a 3 month period, to obtain data on the care of these patients in the hospital. Data were analysed by nursing students as part of their practical training. Some of the results are presented; results were discussed in the classroom and presented and commented in an open meeting with caring personnel (nurses, doctors, volunteers) involved in the care of dying patients.