Hypothyroidism decreases proinsulin gene expression and the attachment of its mRNA and eEF1A protein to the actin cytoskeleton of INS-1E cells

The actions of thyroid hormone (TH) on pancreatic beta cells have not been thoroughly explored, with current knowledge being limited to the modulation of insulin secretion in response to glucose, and beta cell viability by regulation of pro-mitotic and pro-apoptotic factors. Therefore, the effects of TH on proinsulin gene expression are not known. This led us to measure: a) proinsulin mRNA expression, b) proinsulin transcripts and eEF1A protein binding to the actin cytoskeleton, c) actin cytoskeleton arrangement, and d) proinsulin mRNA poly(A) tail length modulation in INS-1E cells cultured in different media containing: i) normal fetal bovine serum - FBS (control); ii) normal FBS plus 1 µM or 10 nM T3, for 12 h, and iii) FBS depleted of TH for 24 h (Tx). A decrease in proinsulin mRNA content and attachment to the cytoskeleton were observed in hypothyroid (Tx) beta cells. The amount of eEF1A protein anchored to the cytoskeleton was also reduced in hypothyroidism, and it is worth mentioning that eEF1A is essential to attach transcripts to the cytoskeleton, which might modulate their stability and rate of translation. Proinsulin poly(A) tail length and cytoskeleton arrangement remained unchanged in hypothyroidism. T3 treatment of control cells for 12 h did not induce any changes in the parameters studied. The data indicate that TH is important for proinsulin mRNA expression and translation, since its total amount and attachment to the cytoskeleton are decreased in hypothyroid beta cells, providing evidence that effects of TH on carbohydrate metabolism also include the control of proinsulin gene expression.

A specific and highly sensitive time-resolved fluoroimmunoassay for human proinsulin

We describe a time-resolved fluoroimmunoassay specific for human proinsulin using a combination of two high-affinity monoclonal antibodies, one against insulin and the other specific for intact proinsulin and for split 65-66 and des 64-65 proinsulin forms. The assay employs only 200 micro liters of serum, with a detection limit of 0.1 pmol/l. The intra-assay variation coefficient was less than 3 percent between 3 and 1000 pmol/l. There was 0 percent cross-reaction with insulin, C-peptide, split 32-33 and des 31-32 proinsulin. Serum concentration of proinsulin was analyzed in 50 subjects during an oral glucose tolerance test (10 non-obese controls, 10 obese controls, 10 subjects with impaired glucose tolerance, 10 patients with type II diabetes meIlitus (DM) and fasting blood glucose (FBG) <140 mg/dl, and 10 patients with type II DM and FBG >150 mg/dl). Mean fasting serum proinsulin levels measured by this assay in non-obese controls (0.84 +/-0.90 pmol/l; 0.1-2.4 pmol/l) were lower than the results reported by her investigators. There was an increase of proinsulin related to obesity and increased glucose levels, suggesting that proinsulin levels increase with insulin resistance.

Effects of prednisolone and dexamethasone on (pro)insulin biosynthesis and insulin release by rat islets of Langerhans.

Administration of either prednisolone or dexamethasone (10 mg/kg body wt/day/oral) to rats for 21 days resulted in inhibition of (pro)insulin biosynthesis and immunoreactive insulin release by isolated islets. A gradual reversal of prednisolone's effect was obtained after exposing islets to increasing glucose concentrations but glucose challenges failed to influence dexamethasone's effect.

Changes in pancreatic beta cell function during growth and reproductive maturity of rats.

An attempt has been made to study the changes, if any, in the efficiency of pancreatic beta cell function at different ages using one experimental model. For this, pancreatic islets were isolated from albino rats of 3, 8, 12 and 36 weeks of age and challenged with various concentrations of glucose (11.0, 16.7 and 22.2 mM). Significantly higher rate of glucose stimulated (pro)insulin biosynthesis was seen in the islets from 3 week old rats as compared to the islets obtained from 8, 12 and 36 week rats. On the other hand, immunoreactive insulin release was observed to be highest from islets of 8 week old rats followed by that of 12, 36 and 3 week old ones.

Estudio sobre el mecanismo de accion del zincum metalico / Study about the action mode of the metallic zincum

El objeto de este estudio es presentar la revision documental de un medicamento homeopatico que se ha usado durante doscientos anos. Se trata de um oligoelemento, el zinc, que en los ultimos dieciocho anos ha sido profundamente estudiado en cuanto al aspecto bioquimico, farmacologico y principalmente la accion toxica en el organismo