Effect of yeast culture on in vitro fermentation of a high-concentrate or high-fiber diet using equine fecal inoculum in a Daisy II incubator.

Two experiments were conducted to evaluate the use of a closed system, fermentation apparatus (Daisy II incubator) and determine the effects of a yeast culture (YC) preparation (Saccharomyces cerevisiae) on in vitro microbial populations, diet digestion, and fermentation patterns in horses. In Exp. 1, 4 mature horses were fed a pelleted concentrate and alfalfa cubes in a 50:50 (%, as-fed) ratio. Fecal samples were taken from each horse to form the inoculum and placed in 4 separate incubation vessels. Twenty nylon bags (10 with 0.25 g and 10 with 0.50 g of the total mixed diet) were placed in each vessel, and in vitro fermentation was carried out for 48 h to determine DM, NDF, and ADF digestibility. In Exp. 2, fecal samples were taken from 4 mature horses consuming either a high-concentrate (HC) or high-fiber (HF) diet. Filter bags containing the HC or HF diet were added to the 4 incubation vessels along with their respective inoculums. Yeast culture was added to 2 of the vessels containing the HC or HF diet, whereas the other 2 vessels served as controls. Vessels were incubated as in Exp. 1 with samples taken at 24 and 48 h. Filter bags were used to determine DM, NDF, ADF, and OM digestibility, whereas vessel fluid was analyzed for lactate, ammonia, VFA, and microbial concentrations. Results of Exp. 1 indicated that DM, NDF, and ADF digestibility were greater (P < 0.05), whereas the corresponding CV was lower (P < 0.05) for the 0.25- vs. the 0.50-g sample size. In Exp. 2, YC tended (P = 0.10) to decrease ammonia concentrations in the HF diet and increased (P < 0.05) acetate production in the HC diet when compared with the control. There were no effects of YC on pH, lactate, or the measured microbial populations, as well as DM, NDF, or ADF digestibility. The results did, however, show that in vitro and in vivo DM digestibility estimates were similar within a diet. Data from Exp. 1 indicated that the 0.25-g sample size provides a more accurate estimate of DM digestibility with less variation. Although YC had little, if any, effect in Exp. 2, the results indicated that the Daisy II incubator does provide valid estimates of total tract DM digestibility in the horse. These data provide further evidence that this process would be an effective and practical means of approximating the digestibility of diets with varying concentrate to forage ratios.

A facet approach to extending the normative component of the theory of reasoned action.

Using facet theory, this study addresses the weak explanatory power of normative influence in theories of reasoned action or planned behaviour. A broad normative construct is hypothesized as being characterized by two facets--social unit and behavioural modality--each of which is examined in relation to recreational drug use. A questionnaire was developed from the facets and administered to undergraduate students. Data (N = 181) were analysed using Smallest Space Analysis (SSA). The results suggest that the facets provide an adequate description of the normative construct and that personal and social normative beliefs, behavioural norms and behavioural intentions can be distinguished empirically. The results also lend partial support to Ajzen's (1988; Ajzen & Fishbein, 1977) principle of compatibility. Implications for how social influence is operationalized and conceptualized are also discussed.

Antisense oligonucleotides as research tools.

The use of antisense oligonucleotides as both research tools and therapeutic molecules has emerged as a powerful alternative to small molecule inhibitors. Antisense oligonucleotides are short pieces of chemically modified DNA designed to hybridize to specific mRNA sequences present in the target gene. The oligonucleotide interaction with the targeted mRNA can lead to inhibition in the translation of the protein encoded by the targeted transcript through a variety of reasonably well-characterized mechanisms (1-3).

Intercellular adhesion molecule-1 suppression in skin by topical delivery of anti-sense oligonucleotides.

We topically applied 20 nucleotide phosphorothioate intercellular adhesion molecule-1 anti-sense oligodeoxynucleotide in a cream formulation. It effectively inhibited tumor necrosis factor-alpha-induced expression of intercellular adhesion molecule-1 in human skin transplanted on severe compromised immunodeficient mice. The effects were concentration dependent, sequence specific, and resulted from reduction of intercellular adhesion molecule-1 mRNA levels in the skin. Intravenous administration of the drug did not show pharmacologic effects, probably due to insufficient drug concentrations in skin. Topical delivery, however, produced a rapid and a significantly higher accumulation of oligodeoxynucleotide in the epidermis and dermis. The results strongly suggest that topically applied anti-sense oligonucleotides can be delivered to target sites in the skin and may be of considerable value in the treatment of psoriasis and other inflammatory skin disorders.

Inhibition of antigen-induced eosinophilia and late phase airway hyperresponsiveness by an IL-5 antisense oligonucleotide in mouse models of asthma.

Chronic airway eosinophilia is associated with allergic asthma and is mediated in part by secretion of IL-5 from allergen-specific Th2 lymphocytes. IL-5 is a known maturation and antiapoptotic factor for eosinophils and stimulates release of nascent eosinophils from bone marrow into the peripheral circulation. An antisense oligonucleotide found to specifically inhibit IL-5 expression in vitro was observed to significantly reduce experimentally induced eosinophilia in vivo, in both the murine OVA lung challenge and allergic peritonitis models. Intravenous administration resulted in sequence-dependent inhibition of eosinophilia coincident with reduction of IL-5 protein levels, supporting an antisense mechanism of action. Potent suppression of lung eosinophilia was observed up to 17 days after cessation of oligonucleotide dosing, indicating achievement of prolonged protection with this strategy. Furthermore, sequence-specific, antisense oligonucleotide-mediated inhibition of Ag-mediated late phase airway hyperresponsiveness was also observed. These data underscore the potential utility of an antisense approach targeting IL-5 for the treatment of asthma and eosinophilic diseases.

Pharmacology of antisense oligonucleotide inhibitors of protein expression.

The dramatic increase in recent years of both the amount and rate of accumulation of novel genomic sequence information has generated enormous opportunities for the development of new classes of drugs. For these opportunities to be fully capitalized upon, investigators must choose molecular targets for drug development that are likely to yield attractive therapeutic profiles. This will require rapid and effective determination of gene functions in multiple cellular settings. The development of antisense oligonucleotides as specific inhibitors of gene expression should allow such determination of gene function. In addition, the antisense oligonucleotides themselves will likely prove useful as drugs. In this review, we discuss some of the issues surrounding the use of antisense oligonucleotides as research tools to help elucidate gene function, and highlight some of the approaches that can be taken to generate and use effective antisense reagents.

In vivo distribution and metabolism of a phosphorothioate oligonucleotide within rat liver after intravenous administration.

In the rat, the liver represents a major site of phosphorothioate oligodeoxynucleotide deposition after i.v. administration. For this reason, we examined the intracellular fate of ISIS 1082, a 21-base heterosequence phosphorothioate oligodeoxynucleotide, isolated from parenchymal and nonparenchymal cell types after systemic dosing using established perfusion and separation techniques followed by CGE. Isolated cells were further fractionated into nuclear, cytosolic and membrane constituents to assess the intracellular localization, distribution and metabolic profiles as a function of time and dose. After a 10-mg/kg i.v. bolus, intracellular drug levels where maximal after 8 hr and diminished significantly thereafter, suggesting an active efflux mechanism or metabolism. Nonparenchymal (i.e., Kupffer and endothelial) cells contained approximately 80% of the total organ cellular dose, and this was equivalently distributed between the two cell types, while the remaining 20% was associated with hepatocytes. Nonparenchymal cells contained abundant nuclear, cytosolic and membrane drug levels over a wide dose range. In contrast, at doses of less than 25 mg/kg, hepatocytes contained significantly less drug with no detectable nuclear-association. Doses at or above 25 mg/kg appeared to saturate nonparenchymal cell types, whereas hepatocytes continued to accumulate drug in all cellular compartments, including the nucleus. Our results suggest that although pharmacokinetic parameters vary as a function of hepatic cell type, significant intracellular delivery can be readily achieved in the liver after systemic administration.

Role and function within managed care organizations: opportunities for nurse managers.

Health care systems are facing rapid changes, moving from a fee-for-service environment to managed care. These changes are impacting nursing, eliminating current jobs and creating new opportunities in the field of managed care. Nurses must take a critical look at their current positions and determine what their contributions will be to improving the system of care. It is imperative that nurses move to leadership positions in managed care to represent the needs of patients and providers at the boardroom table. The purpose of this article is to give an insider's view of traditional and nontraditional roles for nurses within managed care organizations, and to challenge nurses to seize the opportunity to participate in designing the health care delivery system of the future.

New skills sets needed to navigate managed care.

Nurse managers in the late 1990s and the 21st century may find themselves working for a managed care organization, and certainly will find themselves employed in a managed health care environment. The roles and opportunities are many for managers who have the right combination of skills and knowledge. Understanding managed care language and strategies in synergy, with a creative leadership approach, enables nurse managers to move successfully from acute care to the new world now unfolding.

Surface-relief diffraction gratings recorded by multiple-beam coherent phase exposure.

Surface-relief hexagonal-array diffraction gratings have been produced by three-beam coherent exposure. Collimated light was used in an attempt to produce a uniform relief profile over the total area of a 7.6-cm plate coated with a positive photoresist. The resulting gratings were reproduced in nickel by an electroforming process and analyzed by atomic force microscopy. The topography of the gratings was found to be that predicted by theory. The results obtained show that the gratings were of uniform profile over their total area.

Long-term history of chesapeake bay anoxia.

Stratigraphic records from four sediment cores collected along a transect across the Chesapeake Bay near the mouth of the Choptank River were used to reconstruct a 2000-year history of anoxia and eutrophication in the Chesapeake Bay. Variations in pollen, diatoms, concentration of organic carbon, nitrogen, sulfur, acid-soluble iron, and an estimate of the degree of pyritization of iron indicate that sedimentation rates, anoxic conditions and eutrophication have increased in the Chesapeake Bay since the time of European settlement.

Siderophore electrochemistry: relation to intracellular iron release mechanism.

Previous studies have shown that there is a major difference between the iron release mechanism of enterobactin, a catechol-based siderophore, and that of the hydroxamate-based siderophores such as ferrichrome. For ferric enterobactin there is an esterase that hydrolyzes the ligand during iron release. In contrast, iron is released by the hydroxamate-based siderophores and the ligands are reused in subsequent iron transport. It has been suggested that release of iron by hydroxamates occurs by reduction to the ferrous complex, a process that does not occur for ferric enterobactin. Cyclic voltammograms of ferrichrome A and ferrioxamine B exhibit reversible one-electron waves with pH-independent formal potentials (Ef-vs. the normal hydrogen electrode) -446 and -454 mV, respectively, within the range of physiological reductants. Ferric enterobactin also shows a reversible one-electron wave (at pH greater than 10) with Ef = -986 mV vs. the normal hydrogen electrode. From the pH dependence of this potential we estimate a reduction potential of -750 mV at pH 7. In sharp contrast to the value for the ferric hydroxamates, this value is well below the range of physiological reducing agents. The results demonstrate that the observed hydrolysis of enterobactin is a necessary prerequisite to in vivo release of iron from the siderophore via ferric ion reduction.