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R F Power

Publications and source records attributed to R F Power.

At least 37 records · Page 2Linked to original sources

Technical note: detection and quantification of supplemental fungal beta-glucanase activity in animal feed.

Selected hydrolytic enzymes are added to animal feeds in order to degrade specific antinutritional factors and(or) to increase availability of certain components of feedstuffs to the animal. A method is described that allows detection and quantification of beta-glucanase activity in complex feedstuffs. The method is based on radial diffusion of an enzyme-containing feed extract through an agar gel in which lichenan substrate (a relatively inexpensive glucan of mixed beta 1-->4 and beta 1-->3 linkages) has been dissolved. A linear relationship between the diameter of the zone of substrate hydrolyzed and the log of enzyme activity present was observed. The assay described is technically straightforward and requires no specialized equipment. At typical commercial inclusion levels (1 kg/t), the activity of a supplemental beta-glucanase, added to feed in a commercial mill was determined by averaging several measurements, with a precision of +/- 4%, variation between individual readings of +/- 11.3% (SD), and recovery of 109%. By using high-concentration feed extracts, the method was sensitive enough to detect background and(or) supplemental beta-glucanase activities as low as .05 kg/t supplement equivalent. This method allows consumers, producers, and regulatory authorities to measure the activity of beta-glucanase in feed at commercial inclusion levels and, hence, study the effects of processes such as pelleting and extrusion on such supplements.

Animal Feed↗

Analysis of cytokine mRNA expression in syngeneic islet grafts of NOD mice: interleukin 2 and interferon gamma mRNA expression correlate with graft rejection and interleukin 10 with graft survival.

The injection of complete Freund's adjuvant into diabetic nonobese diabetic (NOD) mice at the time of syngeneic islet transplantation prevents monocytic/lymphocytic cell infiltration into the islet graft, Beta-cell destruction, and autoimmune diabetes recurrence. We have used semiquantitative reverse transcriptase-polymerase chain reaction analysis to examine and compare cytokine mRNA expression profiles in islet grafts from complete Freund's adjuvant-injected and control NOD mice. Interleukin 10 mRNA expression was significantly increased whereas interleukin 2 and interferon gamma mRNA levels were significantly decreased in islet grafts from complete Freund's adjuvant-injected mice compared to control mice. Levels of mRNA for interleukin 1 beta, interleukin 4, and tumour necrosis factor alpha were not significantly different in islet grafts from complete Freund's adjuvant-injected and control mice. These findings suggest that a Th1 subset of lymphocytes and their cytokine products, interleukin 2 and interferon gamma, may be involved in the rejection of syngeneic islet grafts and diabetes recurrence in NOD mice, and that the protective effect of complete Freund's adjuvant may result from the induction of interleukin 10 production and consequent down-regulation of Th1 cells and cytokines in the islet graft.

Animals↗

Dysplastic tubular epithelium in "normal" kidney associated with renal cell carcinoma.

Intratubular epithelial dysplasia (IED) of the renal tubules has not been fully described in human renal cell carcinoma (RCC). This lesion has been found in male Syrian hamsters exposed to estrogens. One article reports IED in human kidney showing nephrosclerosis and RCC. We examined "normal" kidney tissue adjacent to 110 cases of RCC in an attempt to identify possible precursor lesions. There were 73 male and 37 female patients (M/F = 2:1). The ages ranged from 27 to 86 years (median 64 years). IED was identified in 30 cases. The lesions consisted of foci of crowded tubular epithelium with large, vesicular nuclei two to three times the size of nuclei of benign tubular cells with eosinophilic macronucleoli. The tubules were occasionally filled with dysplastic cells mimicking carcinoma in situ. The lesions were predominantly cortical and periglomerular. They either were subtle and focal or, less commonly, involved tubules diffusely. Eighteen of the 73 male patients (24%) had these lesions compared with 12 of 37 female patients (32%). They were more usually seen in the clear cell (21 of 66) and sarcomatoid (three of four) variants of RCC than in the oncocytic/granular cell (four of 25) or tubulopapillary (two of 14) variants. One case of collecting duct RCC showed no evidence of IED. Immunohistochemical assessment of 20 dysplastic and 20 nondysplastic lesions with their adjacent RCC for cytokeratin, vimentin, cathepsin-D, and epidermal growth factor receptors was inconclusive. Our findings suggest that IED associated with RCC might represent previously unrecognized precursor lesions along the spectrum ranging from dysplasia to frank carcinoma. The biological significance of these lesions, their preponderance in women, and the phenotypic and genotypic characteristics require further investigation.

Adult↗

An ultrafiltration method for the removal of interfering agents and its application to the determination of free ammonia in solutions of oxystarch by the Berthelot reaction method.

Oxystarch was chosen as a model compound for studying biological ammonia-sequestering systems. Ammonia was determined by use of an ion-selective electrode, by L-glutamate dehydrogenase (L-GDH), and by two different Berthelot procedures, in the presence and absence of oxystarch. In the presence of total oxystarch the Berthelot method, particularly when low concentration reagents were used, detected significantly less (P < 0.10) free ammonia than either L-GDH or ion-selective electrode methods. A 0.5-kDa molecular weight cutoff sample ultrafiltration step was added prior to analysis by L-GDH and Berthelot procedures. To facilitate complete removal of oxystarch by the ultrafiltration step, oxystarch was dialyzed before use, yielding a high-molecular-weight fraction (> 1 kDa). Removal of high-molecular-weight oxystarch species and bound ammonia by ultrafiltration of samples prior to assay completely negated discrepancies between ammonia levels measured by L-GDH and both Berthelot methods. The correlation of the levels of measured ammonia, as determined by L-GDH and Berthelot methods, in mixtures with high-molecular-weight oxystarch was significantly improved by the addition of the sample ultrafiltration step. Improved correlation of results from such fundamentally different methods demonstrates the removal of interfering agents as well as the nonperturbatory nature of the improved procedure. The addition of such an ultrafiltration step may be applied to the determination of ammonia by the otherwise interference-prone Berthelot assay in mixtures with any interfering macromolecules, without the inconvenience or potential variabilities associated with distillation or diffusion procedures.

Ammonia↗

Prevention of diabetes and induction of non-specific suppressor cell activity in the BB rat by an immunomodulatory azaspirane, SK&F 106610.

Immunomodulatory azaspirane compounds have immunosuppressive activity in animal models of autoimmune disease such as adjuvant-induced arthritis and experimental autoimmune encephalomyelitis. The mechanism of action of azaspiranes appears to be the induction of antigen non-specific (natural) suppressor cell activity. In this study, we tested the azaspirane, SK&F 106610 in an animal model of autoimmune (type 1) diabetes, the BB rat. Oral administration of SK&F 106610 (15 mg/kg/day) to diabetes-prone BB rats, from age 30 days, significantly decreased diabetes incidence at 100 days from 80% (24 of 30 control rats) to 32% (10 of 31 drug-treated rats, P < 0.001). Protection from diabetes by SK&F 106610 was accompanied by decreased lymphocytic infiltration of the pancreatic islets (insulitis). No changes occurred in splenic T cell, B cell or macrophage subsets, or in proliferative responses to the mitogens lipopolysaccharide and concanavalin A (Con-A). Cell mixing experiments in vitro, however, revealed increased antigen non-specific suppressor activity (suppression of splenic lymphoproliferative response to Con-A) in spleens of SK&F 106610-treated rats. The suppressor cell activity was enriched in a low density fraction of splenic cells relatively depleted of T cells, B cells, macrophages and natural killer cells. These results indicate that the azaspirane compound, SK&F 106610 can prevent insulitis and autoimmune diabetes in BB rats and that these actions may be related to the activation of non-specific (natural) suppressor cells.

Administration, Oral↗

Enzymes in the animal-feed industry.

The increasing economic pressures currently being placed upon animal producers demand more-efficient utilization of low-grade feedstuffs. In addition, consumer awareness and new legislation require that any increase in animal production cannot be achieved via growth-promoting drugs or other chemical substances. One increasingly popular approach to this problem is to supplement animal diets with hydrolytic enzymes in an attempt to aid the digestion and absorption of poorly available nutrients, or to remove antinutritional factors from the diet. Concerns raised by this practice include the ability of such enzymes to survive processing temperatures and even the animals' digestive tract.

Animal Feed↗

Lazaroid antioxidant reduces incidence of diabetes and insulitis in nonobese diabetic mice.

Oxygen free radicals have been implicated as mediators of pancreatic islet beta cell damage in autoimmune, insulin-dependent diabetes mellitus (IDDM). We examined the effects of a potent antioxidant compound, U78518F, on the incidence of diabetes and pancreatic islet infiltration with lymphocytes and monocytes (insulitis) in the non-obese diabetic (NOD) mouse, an animal model for human IDDM. U78518F, administered to NOD mice from age 8 to 30 weeks, produced a dose-dependent decrease in diabetes incidence, from 89% (16/18 control NOD mice) to 61% (11/18 NOD mice treated with 2 mg/kg/day U78518F, p < 0.05) and 44% (8/18 NOD mice treated with 16 mg/kg/day U78518F, p < 0.005). Protection against diabetes was associated with significant decreases in pancreatic insulitis and prevention of beta cell loss in the U78518F-treated NOD mice. These results suggest that antioxidants may have therapeutic application in attempts to prevent immune-mediated islet beta cell damage and IDDM.

Animals↗

Combination therapy with an antioxidant and a corticosteroid prevents autoimmune diabetes in NOD mice.

Oxygen free radicals have been implicated as mediators of pancreatic islet beta cell damage in autoimmune, insulin-dependent diabetes mellitus (IDDM). In this study, we show that the antioxidant, probucol, produced only a small decrease in diabetes incidence in nonobese diabetic (NOD) mice, an animal model for human IDDM. However, combination of probucol with the antiinflammatory corticosteroid, deflazacort, produced an early synergistic effect, delaying diabetes onset by 3 weeks, and a later additive effect, decreasing diabetes incidence from 68% (17 of 25 mice) to 23% (6 of 26 mice, p < 0.005). Protection against diabetes by the combination of probucol and deflazacort was associated with a significant decrease in pancreatic islet infiltration by macrophages/lymphocytes (insulitis) and prevention of islet beta cell loss.

Animals↗

New insights into activation of the steroid hormone receptor superfamily.

For many years the prevailing view of how steroid hormone receptors exert their effects on gene transcription has been that these intracellular receptor proteins, upon association with their specific cognate ligands, undergo a transformation to a state where they are capable of interacting with chromatin and regulating the transcription of specific genes. It has become implicit dogma in the field of biochemical endocrinology that receptor activation is absolutely dependent upon this ligand-binding event. However, recent studies described here by Bert O'Malley and colleagues have shown that certain members of the steroid hormone receptor superfamily can be activated in a totally ligand-independent manner by a cell membrane receptor agonist, the neurotransmitter dopamine.

Animals↗

Mechanisms of complete Freund's adjuvant protection against diabetes in BB rats: induction of non-specific suppressor cells.

We have previously reported that a single injection of complete Freund's adjuvant (CFA) can prevent diabetes appearance in diabetes-prone (DP) BB rats. In this study, we investigated further the mechanism of CFA-induced protection from diabetes. We found that adoptive transfer of splenic cells from CFA-treated DP rats into young DP rats protected the latter from diabetes development. This suggested that CFA-induced protection from diabetes resulted from activation of regulatory (suppressor) cells. Cell mixing experiments in vitro indicated that CFA activated splenic cells with antigen-nonspecific suppressor activity (suppression of lymphoproliferative responses to lipopolysaccharide and to allogeneic splenic cells). Fractionation of splenic cells on Percoll revealed that the suppressor activity resided in low density cells relatively depleted of T-cells, B-cells, macrophages and NK cells. These results suggest that non-specific (natural) suppressor cells in CFA-treated BB rats may be responsible for suppressing autoimmune responses and preventing insulitis and diabetes development.

Animals↗

Differential modes of activation define orphan subclasses within the steroid/thyroid receptor superfamily.

We report that three orphan receptors, hERR1, hERR2, and hTR2, members of the steroid/thyroid receptor (SR/TR) superfamily, can be activated by different ligand-independent pathways. hERR1 and hERR2 exhibit constitutive activity in the absence of exogenously added ligands. Furthermore, this constitutive activity is localized in the carboxy terminal domain of both receptors and can be transferred to other members of this superfamily using domain switch strategies. In addition, we show that hERR1 can remain constitutively active in the less evolved eukaryotic cell Saccharomyces cerevisiae. In contrast, hTR2 is not constitutively active. However, a chimera of hTR2 can be activated in a ligand-independent manner through a signal transduction pathway initiated at the cell membrane by the neurotransmitter dopamine. Like hERR1 and hERR2, hTR2 is ligand-independently activated through its carboxy terminal domain. Together, these results suggest the existence of emerging subgroups within the SR/TR superfamily that can regulate gene expression through different modes of activation.

Base Sequence↗

Dopaminergic and ligand-independent activation of steroid hormone receptors.

The current view of how steroid hormone receptors affect gene transcription is that these receptors, on binding ligand, change to a state in which they can interact with chromatin and regulate transcription of target genes. Receptor activation is believed to be dependent only on this ligand-binding event. Selected steroid hormone receptors can be activated in a ligand-independent manner by a membrane receptor agonist, the neurotransmitter dopamine. In vitro, dopamine faithfully mimicked the effect of progesterone by causing a translocation of chicken progesterone receptor (cPR) from cytoplasm to nucleus. Dual activation by progesterone and dopamine was dissociable, and a serine residue in the cPR was identified that is not necessary for progesterone-dependent activation of cPR, but is essential for dopamine activation of this receptor.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗

Dopamine activation of an orphan of the steroid receptor superfamily.

The chicken ovalbumin upstream promoter transcription factor (COUP-TF) is a member of the steroid receptor superfamily and participates in the regulation of several genes. While a number of functions have been ascribed to COUP-TF, no ligand or activator molecule has been identified, and thus it is classified as one of a group of orphan receptors. Activation of COUP-TF by physiological concentrations of the neurotransmitter dopamine was observed in transient transfection assays. Treatment of transfected cells with the dopamine receptor agonist alpha-ergocryptine also activated COUP-dependent expression of a reporter gene. COUP-TF that contained a deletion in the COOH-terminal domain was not activated by these compounds. These observations suggest that dopamine may be a physiological activator of COUP-TF.

8-Bromo Cyclic Adenosine Monophosphate↗

Heterogeneous binding sites for alpha-atrial natriuretic peptide in human umbilical cord and placenta.

The distributions of atrial natriuretic peptide (ANP) clearance receptors in human umbilical cord and placenta were investigated by using des[Gln18,Ser19,Gly20,Leu21,Gly22]ANP-(4-2 3) (C-ANP) as a specific ligand of this receptor to displace bound alpha-125I-labeled ANP. alpha-125I-ANP bound reversibly to umbilical venous and arterial intima and to fetal placental and maternal decidual tissues, with dissociation constants of 1.24 +/- 0.51, 0.58 +/- 0.19, 1.86 +/- 0.51, and 1.07 +/- 0.25 mM, respectively. Binding was reversed by 1 microM unlabeled alpha-ANP but not by unrelated peptides such as gastrin. The 1 microM C-ANP displaced bound alpha-125I-ANP from the intima of umbilical artery but not that of the vein. The specific reversible binding of alpha-125I-ANP to placental decidua but not to fetal placenta was also displaced by 1 microM C-ANP. Therefore high-affinity binding sites for alpha-ANP on the intima of the umbilical artery and on placental decidua differ from those on the umbilical vein and on fetal placental tissue; the binding sites of the intima of the umbilical artery and of the decidua in humans are consistent with clearance receptors for alpha-ANP.

Adult↗

High level expression of a truncated chicken progesterone receptor in Escherichia coli.

Using a novel Escherichia coli system we have successfully overexpressed a region of the chicken progesterone receptor which encodes both the DNA- and hormone-binding domains. The expression system produces the truncated receptor fragment as an in-frame fusion with ubiquitin. This strategy greatly enhances both the solubility and stability of fusion proteins expressed in E. coli. Synthesis has been further improved by induction of the lambda PL promoter with nalidixic acid at low growth temperatures (less than or equal to 30 degrees C) rather than use of conventional heat induction protocols. We can produce 10 mg of receptor fragment/liter of cells using this system, and we estimate that at least 0.3 mg of this receptor material is biologically active, as assessed by DNA-binding and hormone-binding assays. Receptor produced in this manner is almost indistinguishable from authentic oviduct progesterone receptor using the criteria of hormone-binding specificity and affinity and binding to a progesterone response element. This expression system offers a cheap convenient method for the production of mg amounts of biologically active derivatives of progesterone receptor for biochemical studies.

Animals↗

Persistence of intrinsic neurones and possible phenotypic changes after extrinsic denervation of human respiratory tract by heart-lung transplantation.

Respiratory tract nerves have cell bodies outside (sensory, sympathetic) and inside (parasympathetic) the organ and contain bioactive peptides. These include calcitonin gene-related peptide and tachykinins (sensory nerves), vasoactive intestinal polypeptide (parasympathetic nerves), and neuropeptide with tyrosine (sympathetic nerves). Because transplantation interrupts the extrinsic nerve supply to the tissues, we have examined transplanted human respiratory tracts (n = 11) removed at retransplantation 2 to 42 months after the primary transplant in order to determine whether any nerves and peptide synthesis persist. As controls to establish nerve distribution in human respiratory tract, tissues were obtained from 10 lung resections and five autopsies. Cryostat sections were immunostained to demonstrate the general neural marker PGP 9.5, neuropeptides, and the catecholamine-synthesizing enzyme tyrosine hydroxylase. Nerves immunoreactive for PGP 9.5 were detected in all transplanted tissues. They were fewer in number overall than in control tissue, significantly so in epithelium of trachea and bronchus where they were present sparsely in only three cases. Nerves immunoreactive for tyrosine hydroxylase were significantly fewer in the transplants. Peptide-immunoreactive nerves were also reduced in number in the transplants, except for vasoactive intestinal polypeptide, which was only significantly changed in blood vessels in the lung. Ganglion cells immunoreactive for tyrosine hydroxylase and neuropeptide with tyrosine were seen in the transplanted tissues in five cases, but never in the control tissues. We conclude that whereas some nerves and neuropeptide synthesis persist after extrinsic pulmonary denervation, potentially significant changes also occur, including the appearance in intrinsic parasympathetic neurones of immunoreactivity for a catecholamine-synthesizing enzyme and a peptide normally found in sympathetic nerves.

Adult↗