RESUMEN
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.
Asunto(s)
Colitis/tratamiento farmacológico , Calostro/química , Proteínas de la Leche/farmacología , Leche/química , Animales , Australia , Enfermedad Crónica , Colitis/inducido químicamente , Citocinas/metabolismo , Productos Lácteos , Sulfato de Dextran/efectos adversos , Modelos Animales de Enfermedad , Glucolípidos/farmacología , Glicoproteínas/farmacología , Lactoferrina/farmacología , Ácidos Linoleicos Conjugados/farmacología , Gotas Lipídicas , Masculino , Ratones , Ratones Endogámicos BALB C , Óxido Nitroso/metabolismo , Osteopontina/farmacología , Ribonucleasa Pancreática/farmacología , Proteína de Suero de Leche/farmacologíaRESUMEN
PURPOSE: Chemotherapy-induced diarrhoea (CID) has a significant impact. A medicinal food product (ReCharge) containing iron-saturated lactoferrin and anhydrous milk fat reduces the detrimental effects of chemotherapy on the gut in animals. We report results of a randomised blinded placebo-controlled phase IIb trial investigating the efficacy and safety of ReCharge in preventing CID. METHODS: Eligible patients were adults due to start the first cycle of a 2- or 3-week-cycle chemotherapy regimen, had an Eastern Cooperative Oncology Group (ECOG) status of 3 or less, had adequate haematological, liver and renal function and provided written informed consent. Patients (197) were randomised to ReCharge or placebo. They consumed 100-g study product for 2 weeks before and 6 weeks after starting chemotherapy, completed daily diaries for 8 weeks and attended clinic visits until 12 weeks (2-week cycles) or 14 weeks (3-week cycles). The primary outcome was days with CID. RESULTS: The mean number of days with diary-recorded CID was marginally but not statistically significantly lower on ReCharge than placebo (-2.0, 95 % CI (-4.7 to 0.7), p = 0.2). The proportion reporting diarrhoea in the previous cycle at the clinic visit was 30 % lower (p = 0.012) on ReCharge. Missing diary data may have contributed to the discrepancy. No significant differences were found in quality of life or other adverse events. CONCLUSIONS: We found no clear evidence that ReCharge reduced CID as measured by patient self-report diary. The converse finding of benefit as recorded at clinic visits and incomplete adherence to diary completion indicates that further research is required into methods for measuring CID.
Asunto(s)
Antidiarreicos/uso terapéutico , Antineoplásicos/efectos adversos , Diarrea/inducido químicamente , Diarrea/prevención & control , Helados , Adulto , Anciano , Antineoplásicos/uso terapéutico , Diarrea/tratamiento farmacológico , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neoplasias/tratamiento farmacológico , Placebos , Calidad de Vida , AutoinformeRESUMEN
The maintenance of the mechanical integrity of the skeleton depends on bone remodeling, the well-coordinated balance between bone formation by osteoblasts and bone resorption by osteoclasts. The coupled action of osteoblasts and osteoclasts is regulated by the action of many local and circulating hormones and factors as well as central regulation by a neurological mechanism. We have previously shown that lactoferrin can promote bone growth. At physiological concentrations, lactoferrin potently stimulates the proliferation and differentiation of primary osteoblasts and acts as a survival factor. Lactoferrin also affects osteoclasts, potently inhibiting their formation. In vivo, local injection of lactoferrin results in substantial increases in bone formation and bone area. In a critical bone-defect model in vivo, lactoferrin was also seen to promote bone growth. The mitogenic effect of lactoferrin in osteoblast-like cells is mediated mainly through low-density lipoprotein-receptor protein-1 (LRP1), a member of the low-density lipoprotein-receptor-related proteins that are primarily known as endocytic receptors; however, LRP1 is not necessary for the anti-apoptotic actions of lactoferrin. Lactoferrin also induces the activation of p42/44 mitogen-activated protein kinase (MAPK) signalling and the PI3-kinase-dependent phosphorylation of Akt in osteoblasts. In this study, we examined other properties of lactoferrin and the way they affect osteogenic activity. The degree of glycosylation, iron-binding, and the structure-activity relationships indicate that lactoferrin maintains osteogenic activity in deglycosylated, holo, and apo forms, and in with various small fragments of the molecule. These data suggest that lactoferrin signals through more than 1 membrane-bound receptor to produce its anabolic skeletal effects, and that it signals through diverse pathways. We conclude that lactoferrin might have a physiological role in bone growth and healing and a potential therapeutic role as an anabolic factor in osteoporosis.
Asunto(s)
Huesos/metabolismo , Lactoferrina/química , Lactoferrina/farmacología , Secuencia de Aminoácidos , Animales , Bovinos , Relación Dosis-Respuesta a Droga , Glicosilación , Humanos , Hierro/metabolismo , Lactoferrina/metabolismo , Ratones , Datos de Secuencia Molecular , Osteoblastos/efectos de los fármacos , Ratas , Proteínas Recombinantes/farmacología , Relación Estructura-ActividadRESUMEN
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.
Asunto(s)
Caseínas/análisis , Caseínas/aislamiento & purificación , Cromatografía Líquida de Alta Presión/métodos , Proteínas de la Leche/análisis , Proteínas de la Leche/aislamiento & purificación , Fragmentos de Péptidos/análisis , Fragmentos de Péptidos/aislamiento & purificación , Peptonas/análisis , Peptonas/aislamiento & purificación , Poliestirenos/química , Aminoácidos/análisis , Animales , Bovinos , Cromatografía de Afinidad , Cromatografía por Intercambio Iónico , Reproducibilidad de los Resultados , Proteína de Suero de LecheRESUMEN
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.
Asunto(s)
Diabetes Mellitus Experimental/enzimología , Inositol/metabolismo , Animales , Encéfalo/enzimología , Masculino , Mio-Inositol-1-Fosfato Sintasa/metabolismo , Oxigenasas/metabolismo , Fosfatidilinositoles/metabolismo , Fosfotransferasas/metabolismo , Ratas , Nervio Ciático/enzimología , Distribución TisularRESUMEN
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.
Asunto(s)
Diabetes Mellitus Experimental/dietoterapia , Inositol/uso terapéutico , Conducción Nerviosa , Nervios Periféricos/análisis , Animales , Cromatografía de Gases , Diabetes Mellitus Experimental/fisiopatología , Inositol/administración & dosificación , Inositol/análisis , Lípidos/análisis , Masculino , Neuronas Motoras , Nervios Periféricos/fisiopatología , RatasRESUMEN
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.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Inositol/metabolismo , Metabolismo de los Lípidos , Enfermedad Aguda , Animales , Metabolismo de los Hidratos de Carbono , Inositol/sangre , Masculino , Ratas , Distribución TisularRESUMEN
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.