In vitro characterization of the effects of chronic insulin stimulation in mouse 3T3-L1 and human SGBS adipocytes.

Hyperinsulinemia is the hallmark of the development of insulin resistance and precedes the diagnosis of type 2 diabetes. Here we evaluated the effects of prolonged exposure (≥4 days) to high insulin doses (150 nM) in vitro in two adipose cell types, mouse 3T3-L1 and human SGBS. Chronic insulin treatment significantly decreased lipid droplet size, insulin signalling and insulin-stimulated glucose uptake. 3T3-L1 displayed an increased basal glucose internalization following chronic insulin treatment, which was associated with increased GLUT1 expression. In addition, both cells showed increased basal lipolysis. In conclusion, we report the effects of prolonged hyperinsulinemia in 3T3-L1 and SGBS, highlighting similarities and discrepancies between the cell types, to be considered when using these cells to model insulin-induced insulin resistance.

Corticosterone alters materno-fetal glucose partitioning and insulin signalling in pregnant mice.

Glucocorticoids affect glucose metabolism in adults and fetuses, although their effects on materno-fetal glucose partitioning remain unknown. The present study measured maternal hepatic glucose handling and placental glucose transport together with insulin signalling in these tissues in mice drinking corticosterone either from day (D) 11 to D16 or D14 to D19 of pregnancy (term = D21). On the final day of administration, corticosterone-treated mice were hyperinsulinaemic (P < 0.05) but normoglycaemic compared to untreated controls. In maternal liver, there was no change in glycogen content or glucose 6-phosphatase activity but increased Slc2a2 glucose transporter expression in corticosterone-treated mice, on D16 only (P < 0.05). On D19, but not D16, transplacental (3) H-methyl-d-glucose clearance was reduced by 33% in corticosterone-treated dams (P < 0.05). However, when corticosterone-treated animals were pair-fed to control intake, aiming to prevent the corticosterone-induced increase in food consumption, (3) H-methyl-d-glucose clearance was similar to the controls. Depending upon gestational age, corticosterone treatment increased phosphorylation of the insulin-signalling proteins, protein kinase B (Akt) and glycogen synthase-kinase 3ß, in maternal liver (P < 0.05) but not placenta (P > 0.05). Insulin receptor and insulin-like growth factor type I receptor abundance did not differ with treatment in either tissue. Corticosterone upregulated the stress-inducible mechanistic target of rapamycin (mTOR) suppressor, Redd1, in liver (D16 and D19) and placenta (D19), in ad libitum fed animals (P < 0.05). Concomitantly, hepatic protein content and placental weight were reduced on D19 (P < 0.05), in association with altered abundance and/or phosphorylation of signalling proteins downstream of mTOR. Taken together, the data indicate that maternal glucocorticoid excess reduces fetal growth partially by altering placental glucose transport and mTOR signalling.

Intra-aortic balloon counterpulsation in penetrating cardiac trauma.

A case is presented in which the intra-aortic balloon pump (IABP) was used to successfully manage cardiogenic shock in a patient with a cardiac stab wound, not involving a coronary artery.

Percutaneous transluminal coronary angioplasty of anomalous right coronary artery.

Coronary angioplasty of tortuous anomalous coronary arteries can be technically challenging. We describe a successful percutaneous transluminal coronary angioplasty (PTCA) of an anomalous right coronary artery after a failed previous attempt. The anatomic limitations of anomalous right coronary arteries and technical considerations for PTCA are discussed.

Iatrogenic internal mammary artery to coronary vein fistula.

Iatrogenic aortocoronary vein fistula following coronary artery bypass surgery is a rare complication. We describe the first reported case of inadvertent anastomosis of the left internal mammary artery to cardiac vein. The clinical characteristics and consequences as well as the angiographic characteristics of this fistula are described. Precautions that may be taken to prevent this complication are also addressed.

3 alpha-, 7 alpha-, and 12 alpha-OH group specific enzymic analysis of biliary bile acids: comparison with gas-liquid chromatograpy.

3 alpha-Hydroxysteroid dehydrogenase (3 alpha-HSDH) from P. testosteroni, 7 alpha-HSDH (Escherichia coli ATCC No. 29532) and 12 alpha-HSDH (Clostridium group P, strain C48-50, ATCC No. 29733) were used to directly measure 3 alpha-, 7 alpha-, and 12 alpha-OH groups in extracted human bile-rich duodenal aspirates. Twelve samples chosen from widely differing ratios of cholic/chemodeoxycholic/deoxycholic were computed by solving three simultaneous equations. Comparison of these ratios with those obtained by a) thin-layer chromatography and 3 alpha-, 7 alpha-HSDH assays and b) gas-liquid chromatographic analysis showed no significant difference. Addition of known amounts of pure cholic, chenodeoxycholic, deoxycholic, or lithocholic acid to individual bile extracts gave an appropriate yield of 3 alpha-, 7 alpha-, and 12 alpha-OH groups. The direct (non-chromatographic) enzymic method has the advantages of being rapid, convenient, and inexpensive, and thus suitable for clinical use.

Quantitative assay of conjugated and free bile acids as heptafluorobutyrate derivatives by gas-liquid chromatography.

Quantitative analyses of individual bile acids in biological samples are limited by the lengthy multistep preparations necessary. Using heptafluorobutyric acid anhydride in pyridine as derivatizing agent, we reduced several steps to one. Bile acids and their glycine and taurine conjugates form stable heptafluorobutyrate derivatives, climinating the need for deconjugation and preparation of methyl esters. The derivatives have been characterized by mass spectrometry, and optimum reaction yields have been determined. Operating conditions for analyzing the bile acid heptafluorobutyrates by gas-liquid chromatography on various column packings were investigated, and 0.5% QF-1 or 3% OV-255 was found suitable. The bile acid derivatives were identical whether starting with the bile acid or the glycine or taurine conjugates. The procedure was applied to a quantitative analysis of artificial mixtures of bile acids and bile conjugates, and also of human bile. The results compared favorably to those obtained with a 3 alpha- and 7 alpha-hydroxysteroid dehydrogenase fluorimetric method.