Influence of organic chromium supplementation on the performance of beef calves undergoing weaning-related stress / Influência da suplementação com cromo orgânico no desempenho de bezerros de corte submetidos ao estresse da desmama

Dietary chromium supplementation before, during, and after weaning was conducted to evaluate the hypothesis that chromium supplementation could reduce weaning-induced cortisol release in beef calves. We examined the effects of chromium supplementation in 150 crossbred calves (male and female) between five and six months of age. The calves were randomly divided by sex and breed into two equal homogeneous groups (n=75). One group was used as the control, and the other experimental group received supplementation with 0.9mg of chromium carbon-amino-phospho-chelate per 100kg BW. The chromium supplement was mixed with mineral salt for the consumption of 0.1% of BW, and the supplement was administered via creep feeding 60 days before and 60 days after forced weaning. Calves were weighed, and their blood and urine samples were obtained at four time-points: T0 (60 days before weaning), T1 (at weaning), T2 (48 hours after weaning), and T3 (60 days after weaning). Blood samples were used to determine chromium, cortisol, total protein, and albumin concentrations, and urine samples were used to determine urinary creatinine and chromium levels. Cumulative weight gain was higher in calves supplemented with chromium before weaning and during the experiment (P<0.05). In addition, weaning-related stress caused an increase in chromium excretion in the urine, and chromium supplementation reduced stress, which resulted in lower cortisol and total protein levels during weaning.(AU)

O estudo foi realizado para avaliar a hipótese de que a suplementação dietética com cromo antes, durante e após a desmama possa diminuir a concentração de cortisol causado por este processo em bezerros de corte. Para tal, foram utilizados 150 bezerros mestiços, machos e fêmeas, entre cinco e seis meses de idade. Os animais foram divididos randomicamente por sexo e grupo genético em dois grupos homogêneos (n=75), um mantido como controle e outro suplementado com 0,9mg de carboaminofosfoquelato de cromo/100 kg PV misturado a um sal proteinado para ser consumido na base de 0,1% do PV via creep feeding, no decorrer de 60 dias antes e 60 dias após à desmama forçada. Os animais foram pesados e foram coletadas amostras sanguíneas e urinárias no M0 (60 dias antes da desmama), M1 (desmama), M2 (48 horas após a desmama) e M3 (60 dias após à desmama) para determinação de cromo, cortisol, proteína total e albumina no sangue e da concentração urinária de creatinina e cromo. O ganho acumulado de peso foi superior nos bezerros suplementados com cromo antes da desmama e no decorrer de todo o experimento (P<0,05). A suplementação com cromo reduziu os teores de cortisol e de proteína total durante a desmama. O estresse da desmama provocou aumento da excreção de cromo pela urina.(AU)

Evaluation of 6ß-hydroxycortisol, 6ß-hydroxycortisone, and a combination of the two as endogenous probes for inhibition of CYP3A4 in vivo.

An endogenous probe for CYP3A activity would be useful for early identification of in vivo cytochrome P450 (CYP) 3A4 inhibitors. The aim of this study was to determine whether formation clearance (CL(f)) of the sum of 6ß-hydroxycortisol and 6ß-hydroxycortisone is a useful probe of CYP3A4 inhibition in vivo. In human liver microsomes (HLMs), the formation of 6ß-hydroxycortisol and 6ß-hydroxycortisone was catalyzed by CYP3A4, and itraconazole inhibited these reactions with half maximal inhibitory concentration (IC(50))(,u) values of 3.1 nmol/l and 3.4 nmol/l, respectively. The in vivo IC(50,u) value of itraconazole for the combined CL(f) of 6ß-hydroxycortisone and 6ß-hydroxycortisol was 1.6 nmol/l. The greater inhibitory potency in vivo is probably due to circulating inhibitory itraconazole metabolites. The maximum in vivo inhibition was 59%, suggesting that f(m,CYP3A4) for cortisol and cortisone 6ß-hydroxylation is ~60%. Given the significant decrease in CL(f) of 6ß-hydroxycortisone and 6ß-hydroxycortisol after 200-mg and 400-mg single doses of itraconazole, this endogenous probe can be used to detect moderate and potent CYP3A4 inhibition in vivo.

Local and systemic glucocorticoid metabolism in inflammatory arthritis.

BACKGROUND: Isolated, primary synovial fibroblasts generate active glucocorticoids through expression of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1). This enzyme produces cortisol from inactive cortisone (and prednisolone from prednisone). OBJECTIVE: To determine how intact synovial tissue metabolises glucocorticoids and to identify the local and systemic consequences of this activity by examination of glucocorticoid metabolism in patients with rheumatoid arthritis (RA). METHODS: Synovial tissue was taken from patients with RA during joint replacement surgery. Glucocorticoid metabolism in explants was assessed by thin-layer chromatography and specific enzyme inhibitors. RT-PCR and immunohistochemistry were used to determine expression and distribution of 11beta-HSD enzymes. Systemic glucocorticoid metabolism was examined in patients with RA using gas chromatography/mass spectrometry. RESULTS: Synovial tissue synthesised cortisol from cortisone, confirming functional 11beta-HSD1 expression. In patients with RA, enzyme activity correlated with donor erythrocyte sedimentation rate (ESR). Synovial tissues could also convert cortisol back to cortisone. Inhibitor studies and immunohistochemistry suggested this was owing to 11beta-HSD2 expression in synovial macrophages, whereas 11beta-HSD1 expression occurred primarily in fibroblasts. Synovial fluids exhibited lower cortisone levels than matched serum samples, indicating net local steroid activation. Urinary analyses indicated high 11beta-HSD1 activity in untreated patients with RA compared with controls and a significant correlation between total body 11beta-HSD1 activity and ESR. CONCLUSIONS: Synovial tissue metabolises glucocorticoids, the predominant effect being glucocorticoid activation, and this increases with inflammation. Endogenous glucocorticoid production in the joint is likely to have an impact on local inflammation and bone integrity.

Prereceptor regulation of glucocorticoid action by 11beta-hydroxysteroid dehydrogenase: a novel determinant of cell proliferation.

Isozymes of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) act at a prereceptor level to regulate the tissue-specific availability of active glucocorticoids. To examine the effect of this on cell proliferation and differentiation, we have developed transfectant variants of a rat osteosarcoma cell line that express cDNA for 11beta-HSD1 (ROS 17/2.8beta1) or 11beta-HSD2 (ROS 17/2.8beta2). ROS 17/2.8beta1 showed net conversion of cortisone to cortisol whereas ROS 17/2.8beta2 showed only inactivation of cortisol to cortisone. There was no significant difference in glucocorticoid receptor (GR) expression between the different clones. However, in proliferation and differentiation studies, ROS 17/2.8beta2 cells were completely resistant to cortisol. In contrast, ROS 17/2.8beta1 were sensitive to both cortisone and cortisol. Expression of 11beta-HSD1 decreased cell proliferation whereas 11beta-HSD2 increased proliferation. These responses appear to be due to metabolism of endogenous serum glucocorticoids; proliferation of ROS 17/2.8beta1 decreased further with exogenous cortisone or cortisol whereas ROS 17/2.8beta2 were resistant to both compounds. The pro-proliferative effects of 11beta-HSD2 were abrogated by 18beta-glycyrrhetinic acid, an 11beta-HSD inhibitor, and in cells transfected with cDNA encoding inactive 11beta-HSD2. Data indicate that differential regulation of 11beta-HSD1 and 2 (rather than GR expression) is a key determinant of cell proliferation. Dysregulated expression of 11beta-HSD2 may be a novel feature of tumorigenesis.

[The induction of the migration of bone marrow stem elements to the periphery by the cytokines of cortisone-resistant thymocytes]. / Induktsiia migratsii stvolovykh élementov kostnogo mozga na periferiiu tsitokinami kortizonrezistentnykh timotsitov.

We present evidence about the functional activity of factors produced by cortisol-resistant thymocytes. Experiments in vivo have shown that the administration of the supernatant prepared from cortisol-resistant thymocytes leads to a strong stimulation of endogenous colony formation, significantly prolongs survival of sublethally irradiated mice, increases the rate of restoration of the number of thymocytes in the thymus after sublethal irradiation, and contributes to the recovery of the humoral immune response of nude mice to T-dependent antigens. The results obtained suggest that along with other cytokines, cortisol-resistant thymocytes spontaneously produce a chemotactic factor inducing migration of stem cells from the bone marrow to the periphery.

Sulphasalazine therapy in rheumatoid arthritis. A two-year study and follow-up of clinical results.

Sulphasalazine is among the many drugs in the secondary line of antirheumatic efficacy that have both immunosuppressive and auto-inflammatory properties. Twenty-six patients with definite rheumatoid arthritis (RA) were treated for two years with sulphasalazine in a minimum dose of 1 g per day. Four patients presented untoward effects such as: marked physical asthenia, erythema and changes of the dysproteinemia tests which all disappeared when the drug was withdrawn. The first favourable results in the joint inflammation appeared after 4 weeks. Significant clinical and biologic changes were observed at the end of the study thus suggesting the therapeutic value of sulphasalazine in RA.

Further evidence for a role of arachidonic acid in glucocorticoid teratogenic action in the palate.

Arachidonic acid produces a significant reversal of the production of cleft palate by cortisone in the offspring of sensitive strains of mice in vivo. Arachidonic acid in nanogram per milliliter concentrations also produces a significant reversal of the cortisol inhibition of the programmed cell death of the medial edge epithelium of palatal shelves in vitro. This corrective action of arachidonic acid in vitro is significantly blocked by indomethacin at a nanogram per milliliter concentration which selectively inhibits the conversion of arachidonic acid to prostaglandins and/or thromboxanes at the level of cyclooxygenase. These results support the hypothesis that the inhibition of arachidonic acid release and subsequent prostaglandin and/or thromboxane production by glucocorticoids is involved in the teratogenic action of glucocorticoids and demonstrate that one site of this action is the inhibition of epithelial loss.

Inactivation of corticosteroids in intestinal mucosa by 11 beta-hydroxysteroid: NADP oxidoreductase (EC 1.1.1.146).

Activity of the enzyme 11 beta-hydroxysteroid:NADP oxidoreductase (EC 1.1.1.146) in human intestinal mucosa was determined by incubating scraped mucosa with 3H-cortisone and 14C-cortisol; these steroids were then extracted, separated chromatographically, and the radioactivity assayed to determine simultaneously both reductase and dehydrogenase activities. This was the only significant metabolic alteration which the substrate underwent. Only two cases had slight (5 and 13%) reductase activity. In 35 patients, 16 male and 19 female, including seven cases of Crohn's disease, three ulcerative colitis, five diverticulitis, two undergoing surgery for repair of injuries and 18 for carcinoma of colon or rectum, cortisol was converted to cortisone in 15 min with a wide range of values distributed uniformly up to 85% dehydrogenation, with a mean of 42%. When tissue homogenates were fortified with coenzymes, excess NADPH lowered dehydrogenase activity 81%; excess NADP increased dehydrogenase activity 2-fold in three cases. It is possible that a value is characteristic of an individual but perhaps more likely enzyme activity varies with metabolic events involving changes in the coenzyme levels in mucosa, and a random sampling might be expected to yield such a distribution of values. In any event, where activity is high most of the cortisol is inactivated within minutes. It is suggested that synthetic corticoids which escape such metabolic alteration might, except during pregnancy, prove superior in the treatment of conditions such as inflammatory bowel disease.

Vitamin B6 reduces cortisone-induced cleft palate in the mouse.

Administration of Vitamin B6 during gestation to mice on a Vitamin B6-containing diet resulted in a substantial reduction in cortisone-induced cleft palate. Mice maintained on a Vitamin B6-deficient diet demonstrated an increase in the frequency of cortisone-induced cleft palate; this effect was prevented by administration of Vitamin B6. Vitamin B6 inhibited the specific binding of a labeled glucocorticoid to cytosolic receptors from cultured palatal mesenchyme cells. These results indicate that Vitamin B6 reduces the incidence of cortisone-induced cleft palate by altering the binding of glucocorticoids to their cytoplasmic receptors and subsequently nuclear acceptors.

Effects of vitamin A and cortisone on healing of corneal penetrating wounds.

The effect of the administration of vitamin A drops on neutralizing the inhibitory role of cortisone in penetrating corneal wound healing was studied on 24 albino rabbits (48 eyes). The rate of healing of the wound was determined by the fluorescein strain technique and the tensile strength of the affected cornea was determined on the 11th day after the wound was made. It was observed that in the group of eyes in which only cortisone in any form was administered, not only was the rate of healing delayed (with statistical significance) but there was also a statistically significant lowering of the tensile strength of the wound. However, in the eyes in which vitamin A was also administered simultaneously with cortisone, the rate of healing of the wound was enhanced and there was also a statistically significant increase in the tensile strength of wound. Therefore in this study it was confirmed that by simultaneous use of vitamin A drops, the inhibitory role of cortisone is neutralized even in penetrating corneal wounds.

Influence of cobalt (dietary), cobalamins, and inorganic cobalt salts on phenytoin- and cortisone-induced teratogenesis in mice.

Various cobalt-containing agents (cyanocobalamin, sodium cobaltinitrite, and cobaltous chloride), which formerly had been shown to prevent the onset of cleft palate in CF-1 mice injected with cortisone, were studied to determine whether they would afford similar protection against phenytoin. Phenytoin, however, failed to cause cleft palate in the mouse fetus when given to pregnant animals alone; and cortisone, on the contrary, induced this anomaly in the presence of the so-called cobalt antagonists as well as when administered in their absence. It is suggested from these results that high dietary intake of cobalt prevents cleft palate caused by phenytoin challenge and also negates the protective effects associated with the acute administration of cobalt compounds. Therefore, it is concluded that these well-known teratogens inhibit palatal closure in mice by different mechanisms.

The effect of thorotrast and cortisone on renal cortical fibrinolytic activity in the rabbit.

Previous studies have shown that a single injection of endotoxin inhibits renal cortical fibrinolytic activity in the rabbit. This suggests that the initial injection of endotoxin may prepare for the generalized Shwartzman reaction by depletion of cortical fibrinolytic activity. A fibrin slide technic was used to determine whether Thorotrast(R) and cortisone prepare for the generalized Schwartzman reaction by a similar mechanism. Renal cortical fibrinolytic activity was inhibited following Thorotrast injection, but no inhibition could be detected following cortisone injection. This suggests that Thorotrast, like endotoxin, prepares for the generalized Shwartzman reaction by depletion of cortical fibrinolytic activity. Failure to demonstrate inhibition of lytic activity following cortisone injection may indicate that cortisone prepares for the generalized Shwartzman reaction by another mechanism, or that the fibrin slide technic was unable to detect quantitative changes in lytic activity.

Induction of malate dehydrogenase isoenzymes in livers of young and old rats.

The activity of cytoplasmic malate dehydrogenase (EC 1.1.1.27) of livers of young (9-10 weeks) and old (60-70 weeks) rats decreases after adrenalectomy and increases after administration of cortisone to adrenalectomized rats. These changes are significantly lower in old rats. The induction by cortisone is inhibited by actinomycin D. Adrenalectomy decreases and cortisone increases the activity of mitochondrial malate dehydrogenase of young rats, but not of old rats. Cortisone, however, induces both mitochondrial and cytoplasmic malate dehydrogenase of the regenerating liver at both the ages. Thus, impairment of the inducibility of mitochondrial malate dehydrogenase that occurs in old age is repaired in dividing cells. Immunological studies on purified malate dehydrogenase from young and old rats show that the molecular species of the enzyme is apparently the same at both ages. The changes in activities of the enzymes seen in old age may be due to changes in template activity of the corresponding genes, which apparently do not change with age.