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Previous studies have indicated that the pituitary gland may produce free alpha-subunit and small quantities of hCG in addition to other glycoprotein hormones. Since synthesis of holo-hCG requires the presence of both subunits, we have investigated the occurrence in human pituitary of free beta-subunit of hCG, in addition to intact holo-hCG. We processed a pituitary extract by fractionated ammonium sulfate precipitation followed by sequential chromatography on Sephadex G-100 and Ultrogel AcA 44. The fractions obtained were assessed for their reactivities with a panel of polyclonal and monoclonal antibodies specific for holo-hCG, beta-subunit of hCG, alpha-subunit, or hCG/LH. In addition to the expected LH and alpha-subunit, we detected materials which eluted from the column in positions very similar to those of cochromatographed 125I-hCG tracer and hCG-beta (NIH CR123-beta), and which showed immunoreactivity in specific immunoradiometric assays for holo-hCG and hCG-beta, respectively. Holo-hCG and hCG-beta material derived from the urine of a postmenopausal woman showed behaviors on the column similar to the pituitary forms. Both the pituitary holo-hCG- and free hCG-beta-subunit activity could be enriched (approximately 500 times) by affinity chromatography on an hCG antibody-coupled Sepharose column. When subjected to isoelectric focusing in granulated gel holo-hCG and hCG-beta-subunit of pituitary origin were focused in the pI-range of pregnancy hCG and pregnancy hCG-beta-subunit, respectively. Like pregnancy hCG, most (75%) of the pituitary hCG was bound to a column of Con A-Sepharose; however, the Con A-nonbinding hCG fraction (approximately 25%) was much higher than that found in pregnancy hCG. On the basis of immunoreactivities, the content of holo-hCG in our pituitary extract was estimated to be 60 micrograms/g, and that of free beta-subunit 45 micrograms/g; for comparison, LH was approximately 20 mg/g, and free alpha-subunit 1.6 mg/g. In addition, we could demonstrate the presence of both holo-hCG- and free hCG-beta-subunit-like immunoreactivity in NaCl-extracts from single pituitaries of two postmenopausal women. In these studies a second hCG-beta-immunoreactive material eluting far behind the hCG-beta-position was found. Chromatography of purified LH-beta-subunit, which crossreacts 1.56% in the hCG-beta IRMA, yielded an elution pattern clearly distinguishable from that of the hCG-beta-immunoreactive substances.(ABSTRACT TRUNCATED AT 400 WORDS)

BackgroundGLP-1 is an incretine hormone which gets secreted from intestinal L-cells in response to nutritional stimuli leading to pancreatic insulin secretion and suppression of glucagon release. GLP-1 further inhibits gastric motility and reduces appetite which in conjunction improves postprandial glucose metabolism. Additional vasoprotective effects have been described for GLP-1 in experimental models. Despite these vasoprotective actions, associations between endogenous levels of GLP-1 and cardiovascular disease have yet not been investigated in humans which was the aim of the present study.MethodsGLP-1 serum levels were assessed in a cohort of 303 patients receiving coronary CT-angiography due to typical or atypical chest pain.ResultsGLP-1 was found to be positively associated with total coronary plaque burden in a fully adjusted model containing age, sex, BMI, hypertension, diabetes mellitus, smoking, triglycerides, LDL-C (low density lipoprotein cholesterol), hsCRP (high-sensitive C-reactive protein), and eGFR (estimated glomerular filtration rate) (OR: 2.53 (95% CI: 1.12 – 6.08; p = 0.03).ConclusionCirculating GLP-1 was found to be positivity associated with coronary atherosclerosis in humans. The clinical relevance of this observation needs further investigations.

Abstract. The heat shock protein (HSP) 90 chaperone machine involved in numerous oncogenic signaling pathways is overexpressed in cancer cells and is currently being evaluated for anticancer therapy. Neuroendocrine tumors (NETs) of the gastroenteropancreatic (GEP) system comprise a heterogeneous group of tumors with increasing incidence and poor prognosis. Here, we report the antiproliferative effects of the HSP90 inhibitors AUY922 and HSP990 in neuroendocrine tumor cells. Treatment of human pancreatic BON1, bronchopulmonary NCI-H727 and midgut carcinoid GOT1 neuroendocrine tumor cells with increasing concentrations of AUY922 and HSP990 dose-dependently suppressed cell viability. Significant effects on neuroendocrine cell viability were observed with inhibitor concentrations as low as 5 nM. Inhibition of cell viability was associated with the induction of apoptosis as demonstrated by an increase in sub-G1 events and PARP cleavage. HSP90 inhibition was associated with decreased neuroendocrine ErbB and IGF-I receptor expression, decreased Erk and Akt phosphorylation and the induction of HSP70 expression. These findings provide evidence that targeted inhibition of upregulated HSP90 activity could be useful for the treatment of aggressive neuroendocrine tumors resistant to conventional therapy.

Novel neurotrophin-1/B cell-stimulating factor-3 (NNT-1/BSF-3) is a recently cloned gp130 cytokine, acting through the tripartite ciliary neurotrophic factor receptor (CNTFR) alpha/leukemia inhibitory factor receptor (LIFR)/gp130 receptor complex. The aim of the current study was to investigate the role of NNT-1/BSF-3 in corticotroph cell function and further characterize NNT-1/BSF-3 signaling pathways. Using RT-PCR, expression of ciliary neurotrophic factor receptor alpha, leukemia inhibitory factor receptor, and gp130 could be demonstrated in mRNA derived from murine corticotroph AtT-20 cells and murine pituitary tissue. Incubation of AtT-20 cells with 10 ng/ml recombinant human NNT-1/BSF-3 rapidly induced tyrosine-phosphorylation of signal transducer and activator of transcription (STAT)3 and STAT1 at 5 and 10 min. Proopiomelanocortin promoter activity and suppressor of cytokine signaling (SOCS)-3 promoter activity were significantly stimulated by NNT-1/BSF-3 4.0 +/- 0.3- and 5.9 +/- 0.2-fold, respectively. In comparison with untreated control, NNT-1/BSF-3 significantly stimulated ACTH secretion at 24 and 48 h 1.7 +/- 0.2-fold and 1.5 +/- 0.1-fold above baseline. In comparison with mock-transfected cells, stable overexpression of SOCS-3 in AtT-20 cells abolished NNT-1/BSF-3-induced STAT1 and STAT3 phosphorylation and almost completely inhibited STAT-dependent proopiomelanocortin promoter and SOCS-3 promoter activities. In addition, NNT-1/BSF-3-induced ACTH secretion at 48 h was significantly attenuated by SOCS-3 overexpression. In summary, we have shown that NNT-1/BSF-3 is a modulator of corticotroph cell function, which is negatively regulated by SOCS-3. Our data indicate that the activation of the Jak-STAT cascade is essential for corticotroph NNT-1/BSF-3 signaling. Further studies will have to investigate the possible in vivo role of NNT-1/BSF-3 as a neuroimmunoendocrine modulator of hypothalamus-pituitary-adrenal axis stress response.