Human GnRH-secreting cultured neurons express activinbetaA subunit mRNA and secrete dimeric activin A.

OBJECTIVE: To evaluate the expression of activin betaA-subunit mRNA and the secretion of activin A in/from cultured GnRH-secreting neuronal cells cloned from human olfactory epithelium (FNC-B4), which showed biochemical and antigenic properties of GnRH-secreting neurons. DESIGN: FNC-B4 cells were cultured in basal and conditioned media. METHODS: Reverse transcription-polymerase chain reaction (RTR-PCR) evaluated the expression of activin betaA-subunit mRNA. By using a specific ELISA, dimeric activin A concentrations were measured in culture media, in the absence or presence of carvone or forskolin and with different doses of progesterone, GnRH, and estradiol. RESULTS: RT-PCR experiments performed on total RNA isolated from FNC-B4 cells, using specific primers for the activin betaA gene, showed a 787bp DNA band corresponding to the betaA gene. FNC-B4 cells secreted activin A, and the highest accumulation in conditioned medium was achieved after 3h culture: the addition of forskolin, but not of carvone, was able to stimulate the release of activin A from cultured neuronal cells (P<0.01). When progesterone or GnRH was added, a significant accumulation of activin A was observed (P<0.01), while estradiol administration did not significantly affect activin A secretion. CONCLUSION: To date, this is the only study, in an in vitro human model reporting, that GnRH-secreting neuronal cells expressed activin betaA-subunit mRNA, and released dimeric activin A in culture medium. The expression and secretion of activin suggests that in these cells activin A might exert its action by autocrine/paracrine mechanisms

CXCL10 release in cardiopulmonary bypass: An in vivo and in vitro study

Cardiopulmonary bypass (CPB) leads to systemic and cardiac inflammation. Although the intraoperative blood measurement of some inflammatory cytokines has been recognized as an useful tool in clinical setting, postoperative management still represents a major problem; hence, knowledge about additional possible mediator(s) in time of this process would potentially turn in a clinical benefit. CXCL10 has been shown to be involved in cardiac immune-inflammatory processes. Herein, we aimed to investigate whether and how this chemokine could be a possible early mediator and indicator of inflammation development during CPB. Since cardiac, endothelial and immune cells are all sources of CXCL10, our purpose was also to investigate in vitro CXCL10 secretion pattern in time by those cell types. CXCL10 level was higher in CPB patients before surgery as compared to healthy subjects; CXCL10 level raises earliest in serum of CPB patients and in isolated cardiomyocytes under inflammatory stimuli as compared to other cytokines. CXCL10 might represent a critical early factor in mediating systemic and local cardiac inflammatory response in subjects undergoing CPB, offering opportunity for future monitoring or therapeutic interventions