Comparative activities of milk components in reversing chronic colitis.

Inflammatory bowel disease (IBD) is a poorly understood chronic immune disorder for which there is no medical cure. Milk and colostrum are rich sources of bioactives with immunomodulatory properties. Here we compared the therapeutic effects of oral delivery of bovine milk-derived iron-saturated lactoferrin (Fe-bLF), angiogenin, osteopontin (OPN), colostrum whey protein, Modulen IBD (Nestle Healthsciences, Rhodes, Australia), and cis-9,trans-11 conjugated linoleic acid (CLA)-enriched milk fat in a mouse model of dextran sulfate-induced colitis. The CLA-enriched milk fat significantly increased mouse body weights after 24d of treatment, reduced epithelium damage, and downregulated the expression of proinflammatory cytokines and nitrous oxide. Modulen IBD most effectively decreased the clinical score at d 12, and Modulen IBD and OPN most effectively lowered the inflammatory score. Myeloperoxidase activity that denotes neutrophil infiltration was significantly lower in mice fed Modulen IBD, OPN, angiogenin, and Fe-bLF. A significant decrease in the numbers of T cells, natural killer cells, dendritic cells, and a significant decrease in cytokine expression were observed in mice fed the treatment diets compared with dextran sulfate administered mice. The Fe-bLF, CLA-enriched milk fat, and Modulen IBD inhibited intestinal angiogenesis. In summary, each of the milk components attenuated IBD in mice, but with differing effectiveness against specific disease parameters.

Simultaneous separation and quantitation of the major bovine whey proteins including proteose peptone and caseinomacropeptide by reversed-phase high-performance liquid chromatography on polystyrene-divinylbenzene.

A precise, sensitive and reliable RP-HPLC method was developed to enable not only unequivocal determination of alpha-lactalbumin and beta-lactoglobulin in bovine whey samples, but also simultaneous measurement of proteose peptone, caseinomacropeptide, bovine serum albumin and immunoglobulin G. The optimised method on the Resource RPC column allowed separation of the proteins in 30 min and could be applied to the analysis of soluble proteins in a variety of commercial and laboratory whey products. Furthermore, some qualitative information on protein heterogeneity and quality could be derived from the RP-HPLC analyses with additional data available from on-line electrospray mass spectrometry. Within- and between-day repeatability over a wide range of concentrations was excellent (RSD< or =5%) for all proteins except immunoglobulin G and bovine serum albumin where RSD was 7-10%. Analysis of grouped data from whey protein concentrate and whey protein isolate samples gave a limit of detection of < or =0.3% powder mass and a limit of quantitation of < or =1.0% powder mass for all proteins except immunoglobulin G. Limits of detection and quantitation were 0.6% and 2.0%, respectively, for this protein. Quantitative data obtained by the RP-HPLC method compared very favourably with data obtained by alternative methods of whey protein analysis.

Enzymes of myo-inositol and inositol lipid metabolism in rats with streptozotocin-induced diabetes.

Diabetes, with only mild ketosis, was induced in male rats by a single injection of streptozotocin. After 12 weeks the specific activities of enzymes concerned with the metabolism of inositol and of inositol lipids were measured in various tissues. Inositol 1-phosphate synthase (EC 5.5.1.4) was most active in testis and the activity was significantly less in diabetic rats than in controls on a similar diet. Inositol oxygenase (EC 1.13.99.1), which converts myo-inositol into glucuronic acid, was also less active in kidney from diabetic animals. CDP-diacylglycerol-inositol phosphatidyltransferase (EC 2.7.8.11) and phosphatidylinositol 4-phosphate kinase (EC 2.7.1.68) showed decreased specific activities in brain and sciatic nerve of diabetic rats. By contrast the diabetic state did not affect the specific activities of phosphatidylinositol kinase (EC 2.7.1.67) or phosphatidylinositol 4,5-bisphosphate phosphatase (EC 3.1.3.36) in these tissues. The results are discussed in relation to diabetic neuropathy.

Influence of dietary myoinositol on nerve conduction and inositol phospholipids in normal and diabetic rats.

Observations have been made on motor conduction velocity in the tibial nerve of rats given 35% myoinositol in the diet. Comparison between the values before and with up to nine weeks of dosing revealed no alteration in conduction velocity. In such animals, the free myoinositol content in the sciatic nerve was increased; there was no detectable alteration in the lipid inositol concentration. In a second series of experiments, tibial motor nerve conduction velocity in rats with streptozotocin-induced diabetes was compared with conduction velocity in diabetic animals given 1% supplementary dietary myoinositol, and with a control group of nondiabetic rats. Conduction velocity was reduced in the diabetic animals, but no influence from the added dietary myoinositol was detected. No statistically significantly difference in sciatic nerve myoinositol was demonstrated, but the sorbitol and fructose concentrations were increased. Those animals fed the myoinositol supplement had a significantly lower lipid inositol content. The significance of these findings is discussed.

Free and lipid myo-inositol in tissues from rats with acute and less severe streptozotocin-induced diabetes.

Acute diabetes with ketosis was induced in rats by intraperitoneal streptozotocin and also a milder form of diabetes without ketosis by injecting less of the drug. The acutely diabetic rats were killed 72h after injection and the others after either 2 or 13 weeks. Free and lipid myo-inositol was then measured in various tissues and body fluids by g.l.c. of the trimethylsilyl ether. Serum inositol was increased in the acutely diabetic group, whereas liver inositol was decreased. Brain and kidney inositol concentrations were increased in the mildly diabetic animals at 13 weeks and there was a progressive decrease in sciatic-nerve inositol. Lipid inositol of sciatic nerve was decreased in the acutely diabetic group only. Brain lipid inositol concentration was decreased in mild diabetes at 13 weeks. Possible implications of these findings in relation to diabetic neuropathy was discussed.

Studies in the renal handling of insulin in juvenile diabetics.

A technique is described for the accurate radioimmunoassay of insulin in serum and urine. This method was applied to study of renal clearance and excretion of endogenous and exogenous insulin in untreated juvenile diabetics and healthy young adults. There was good agreement between our results for normal adults and previously reported values. In six non-obese juvenile diabetics, urinary insulin clearance values, both basal (fasting) and following glucose loading (entire range 0.03 ml/min to 1.23 ml/min) were similar to those obtained for the adults (entire range 0.17 ml/min to 2.35 ml/min). The basal urinary excretion in these diabetics was generally of the same order of magnitude as that in the normals. The clearance of exogenous insulin, administered for the first time, was also of the same order as that for endogenous insulin. Markedly elevated urinary clearance and excretion of insulin during fasting and non-fasting states was demonstrated in four non-obese juvenile diabetics with no clinical evidence of abnormal proteinuria, though they demonstrated slight to mild clinical dehydration and acidosis compared with the other diabetics studied. Clearance and excretion of exogenous insulin was similarly elevated. This finding could reflect renal tubular dysfunction in these diabetics, and this dysfunction could relate to even the mild degree of dehydration and acidosis found in this study. Endogenous and exogenous insulin clearance in an obese diabetic child was similar to that for the control group.