Porcine dendritic cells generated in vitro: morphological, phenotypic and functional properties.

Despite the central role that dendritic cells (DC) play in immune regulation and antigen presentation, little is known about porcine DC. In this study, two sources of DC were employed. Bone marrow haematopoietic cell-derived DC (BM-DC) were generated using granulocyte-macrophage colony-stimulating factor (GM-CSF) in the presence or absence of tumour necrosis factor-alpha (TNF-alpha). Monocyte-derived DC (Momicron-DC) were generated with GM-CSF and interleukin-4 (IL-4). In both systems, non-adherent cells developed with dendritic morphology, expressing high levels of major histocompatibility complex (MHC) class II. The presence of TNF-alpha increased the BM-DC yield, and enhanced T-cell stimulatory capacity. Both BM-DC and Momicron-DC expressed the pan-myeloid marker SWC3, as well as CD1 and CD80/86, but were also CD14+ and CD16+. The CD16 molecule was functional, acting as a low-affinity Fc receptor. In contrast, the CD14 on DC appeared to differ functionally from monocyte CD14: attempts to block CD14, in terms of lipopolysaccharide (LPS)-induced procoagulant activity (PCA), failed. The use of TNF-alpha or LPS for DC maturation induced up-regulation of MHC class II and/or CD80/86, but also CD14. Allogeneic mixed leucocyte reactions and staphylococcal enterotoxin B antigen presentation assays demonstrated that these DC possessed potent T-cell stimulatory capacity. No T helper cell polarization was noted. Both the BM-DC and the Momicron-DC induced a strong interferon-gamma and IL-4 response. Taken together, porcine DC generated in vitro possess certain characteristics relating them to DC from other species including humans, but the continued presence of CD14 and CD16 on mature and immature porcine DC was a notable difference.

Synthesis of simplified herboxidiene aromatic hybrids.

The syntheses of simplified aromatic hybrids of the novel polyketide herboxidiene (1) are described. One of the hybrids prepared showed significant herbicidal activity against broad-leaved weeds in post-emergent application.

Management of the patient with short bowel syndrome.

Extensive resection of the small bowel results in impaired digestion of macronutrients and malabsorption of nutrients, fluid, electrolytes, and minerals. Gastric acid hypersecretion and alterations in gut hormonal response further contribute to the problem. Diarrhea, dehydration, electrolyte and acid/base abnormalities, and macronutrient and micronutrient deficiencies ensue, and is termed the short bowel syndrome (SBS). Rare disorders, such as essential fatty acid deficiency and D-lactic acidosis, are a greater concern for the SBS patient. These patients' lives are significantly impacted, and they require close monitoring by a medical team knowledgeable about the disease and its nutritional, metabolic, and psychosocial consequences. Immediate therapies are directed toward fluid resuscitation, wound healing, and initiation of early nutrition support. After medical stabilization, multiple nutritional and medicinal therapies are used to aid bowel adaptation and prevent medical crisis. Advanced practice nurses should be knowledgeable about SBS to educate patients and families about this disease, associated therapies and changes in lifestyle, and how to detect and manage acute changes in medical condition.

Intermediate stages in monocyte-macrophage differentiation modulate phenotype and susceptibility to virus infection.

The kinetics of monocyte-macrophage differentiation was analysed using two Swine Workshop Cluster (SWC) CD molecules: SWC1 and SWC9. Myeloid cells were selected by labelling for the common myeloid antigen, SWC3. Confirmation of macrophage identification used acid phosphatase and phagocytosis activities. During differentiation, SWC1 was gradually lost. SWC9 was absent on monocytes but up-regulated early. Consequently, monocytes were SWC1+ SWC9- and macrophages were SWC1- SWC9+. An additional, intermediate, cell population was identified as SWC1+ SWC9+. Size and granularity characteristics mirrored the monocyte, macrophage and intermediate-cell phenotypes. Overall, SWC9 up-regulation was central in macrophage differentiation and dependent on plasma factors. The concomitant loss of SWC1 was independent of these factors, but always associated with mature macrophages. Upon up-regulation of SWC9, the SWC1+ SWC9+ intermediate monocytic cells became susceptible to African swine fever virus infection. These results demonstrate the heterogeneity of monocytic cell differentiation and the importance of these characteristics for interaction with monocytotropic viruses.

Demonstrating APN value in a capitated market.

To be acknowledged as powerful resources in health care reform, APNs must demonstrate their capability to discern and implement cost-effective care. APNs are typically associated with services that do not generate revenue. Therefore, they need to document their value in terms of improved outcomes, reduced costs, improved patient satisfaction, enhanced institutional image, and better health care team efficiency. The savings generated by APN consultation and followup for patients referred for home TPN was examined in Rochester, NY, using a simplified cost analysis and forms. Collection of data does not need to be elaborate. Costs can be bundled and examined within the period, simple data collection methods can be constructed for details of consultations, and community surveys can provide market prices. It is suggested that the analysis follow basic steps: list items, examine costs, collect or estimate costs, list all assumptions, then calculate total month and daily costs.

Phenotype of porcine monocytic cells: modulation of surface molecule expression upon monocyte differentiation into macrophages.

When porcine blood monocytes differentiate in vitro into macrophages, their morphology, as well as side scatter and forward scatter measured by flow cytometry, changed in a manner similar to that with human cells. During this differentiation, the initial high expression of CD molecules on porcine monocytes was down-regulated, with one exception--SWC9. Freshly isolated blood monocytes were SWC9-, but after culture the cells had become SWC9+. Thus, porcine monocytes were characterised as SWC3+SWC9-CD14high; macrophages were SWC3+SWC9+CD14low, the latter also displaying a down-regulation of CD11a/18, and, to a lesser degree, CD44. Both SWC9-CD14high monocytes and SWC9+CD14low macrophages were identifiable in freshly prepared monocytic cells from the spleen. Alveolar macrophages, on the other hand, were dominated by SWC9+CD14low cells, similar in phenotype to in vitro monocyte-derived macrophages. The consequences which these results have for studies on virus infections of porcine monocytic cells are discussed.

The relative density of CD44-positive porcine monocytic cell populations varies between isolations and upon culture and influences susceptibility to infection by African swine fever virus.

African swine fever (ASF) virus has been reported to infect cells of the monocyte family, probably macrophage-like cells, but there is variation in the apparent susceptibility of these cells. We have demonstrated that the phenotype and activity of porcine monocytic cells varies between different isolations and also upon culture. The variation during culture is dependent upon the phenotype of the cells at the time of isolation. As for the susceptibility of porcine monocytes/macrophages to infection by ASF virus, it was seen that this could be related to the variation in cell phenotype and activity. The susceptibility was determined by the relative density of particular subpopulations of cells present. Whilst inflammatory macrophages did not have an apparent role to play, phagocytic activity was influential. Furthermore, the expression of CD44 and the DH59 myeloid cell marker was important, whereas the relevance of MHC Class II expression was variable. Overall, it was concluded that susceptibility to infection required that a culture be dominated by CD44-positive cells which were non-inflammatory, of low phagocytic activity, and characterizable as being of the myeloid (DH59-positive) lineage.

Pharmacology of moclobemide.

Moclobemide is a reversible inhibitor of monoamine oxidase (MAO) with clear preference for the A type (so-called RIMA). The enzyme inhibition shows complex kinetics, and the molecular mechanism of interaction with the enzyme is not yet clear. Moclobemide increases the extracellular concentration of the monoamines in rat brain and decreases the level of their metabolites. Neither a loss nor a cumulation of activity has been observed after chronic treatment. Reversibility of MAO-A inhibition was demonstrated in vitro as well as in vivo. In various animal behavioral models, in particular in a novel model of stress-induced anhedonia, moclobemide was as effective as standard antidepressants. Moclobemide improves cognitive functions that are impaired in experimental situations. A neuroprotective action is seen in rats subjected to transient global ischemia/-hypoxia. Moclobemide lacks anticholinergic and other effects and only slightly increases the pressor effect of orally administered tyramine. Possible links between MAO-A inhibition and the various effects of moclobemide on brain function are discussed.

Relationship between the anti-FMD virus antibody reaction as measured by different assays, and protection in vivo against challenge infection.

The antibody response of cattle after vaccination against foot-and-mouth disease (FMD) virus was monitored using the serum neutralization test (SNT), the sandwich ELISA, liquid-phase ELISA, sandwich competition ELISA, liquid-phase competition ELISA, and the liquid-phase sandwich competition (blocking) ELISA. The competition ELISAs (in particular the "blocking" ELISA) were the most effective at detecting reactivity in these cattle sera. However, 95% of negative sera also competed in the most sensitive ELISA (the "blocking" ELISA) to titres of 1:32 (4% of the sera competed to a titre of 1:128). Comparisons between the different ELISAs, and between these ELISAs and the SNT, demonstrated that the tests were not measuring exactly the same reaction of antibody with FMD virus. With respect to the capacity of animals to resist FMD virus challenge, neither the SNT nor the competition ELISAs were consistently able to identify such animals. The anti-FMD virus antibody titres obtained could be classified into three zones; the "white zone" wherein antibody titres were high and donor animals likely to be protected; the "black zone" wherein antibody titres were low and donor animals likely to be susceptible to infection; the "grey zone" wherein the antibody titres were intermediary and no interpretation could be made with respect to protection. Assays such as ELISA and SNT cannot and do not measure immunological protection; they are a measure of antibody responses and nothing more, and should be interpreted in terms of the "three zone" phenomenon.

Increasing the impact of the clinical nurse specialist through activity in a shared governance organization.

For the CNS frustrated by traditional placement in an organization, participation in a shared governance organization increases opportunities to improve nursing practice and staff satisfaction. CNSs provide nursing expertise and leadership skills for specific committees as they assist staff nurses in meeting committee objectives. The authors also propose the use of a shared governance organization committee structure as a mechanism for CNSs to increase meaningful interaction with staff nurses and influence their professional development. Their model displays four pathways for CNS-staff nurse interaction through shared governance organization activities: examination of the goals of the profession, evaluation of personal needs and goals, assessment of the characteristics of one's work, and transmission of information. The support provided by the CNS in dealing with these issues increases the staff nurses' sense of a "professional self" and, it is postulated, enhances job satisfaction, decision making, and commitment to the organization's goals.

Pharmacological profile of moclobemide, a short-acting and reversible inhibitor of monoamine oxidase type A.

The novel antidepressant moclobemide is a reversible inhibitor of monoamine oxidase (MAO), preferentially of type A. Moclomide was active in three animal models considered predictive for antidepressant activity: 1) it prevented dose-dependently akinesia and blepharospasm induced in mice and rats by Ro 4-1284, a short-acting amine releasing agent. Prevention of akinesia by moclobemide also depended upon the dose of Ro 4-1284. For comparison also, effects of cimoxatone, harmaline, tranylcypromine and clorgyline are presented: 2) in cats, it selectively and dose-dependently suppressed rapid eye movement sleep without disturbing the sleep-wakefulness cycle; and 3) in the behavioral despair test in mice, it decreased the immobility score to a similar degree as amitriptyline or imipramine. In addition, moclobemide potentiated 5-hydroxytryptophan-induced stereotypies in rats with a potency similar to cimoxatone and with a duration of action of less than 24 hr. Moclobemide had almost no effect on the spontaneous behavior in mice, rats, cats and monkeys. Only in higher doses, marginal sedation and slight impairment in motor performance were seen. Moclobemide did not prevent pilcarpine-induced salivation in mice, demonstrating the absence of anticholinergic activity. Blood pressure and heart rate of freely moving, spontaneously hypertensive rats were only slightly decreased for less than 3 hr. Moclobemide moderately potentiated the pressor effect of p.o. tyramine in rats. In conclusion, the reversible MAO inhibitor moclobemide is active in animal models sensitive to all major drugs used in the treatment of depression. In contrast to imipramine-like antidepressants, it lacks anticholinergic activity and it differs from classic MAO inhibitors by potentiating only weakly the pressor effect of p.o. tyramine.

Inhibition of decarboxylase and levels of dopa and 3-O-methyldopa: a comparative study of benserazide versus carbidopa in rodents and of Madopar standard versus Madopar HBS in volunteers.

The combinations of benserazide and levodopa (1:4, Madopar) and of carbidopa and levodopa (1:10 and 1:4, Sinemet) are currently the most effective treatment of Parkinson's disease. In the present comparative study some effects of the peripheral aromatic L-amino acid decarboxylase (AADC) inhibitors benserazide and carbidopa administered alone or in combination with levodopa by the oral route were investigated in two animal species (rat and mouse) and in healthy volunteers. Benserazide is about 10 times more potent than carbidopa as inhibitor of peripheral AADC both in animals and man. Even at relatively high doses (up to 60 mumol/kg p.o.) benserazide is shown in animals to inhibit the decarboxylation of levodopa only in the extracerebral tissues, thus permitting the formation of dopamine in the striatum and in the hypothalamus. As benserazide is the most potent peripheral AADC inhibitor presently available, is well tolerated and relatively nontoxic even when used chronically, it appears to be the peripheral AADC inhibitor of choice for the development of controlled-release formulations in which Dopa is combined with a peripheral AADC inhibitor. When administered to healthy subjects the pharmacokinetics of the new drug delivery system named Madopar HBS (hydrodynamically balanced system) was characterized by lower and delayed plasma peak concentrations but a longer-lasting concentration of Dopa than after Madopar standard. Therefore, this new controlled-release system may reduce the clinical fluctuations occurring in patients with 'wearing-off' and 'on-off' phenomena.

A three-state model of the benzodiazepine receptor explains the interactions between the benzodiazepine antagonist Ro 15-1788, benzodiazepine tranquilizers, beta-carbolines, and phenobarbitone.

The potent benzodiazepine receptor ligands beta-carboline-3-carboxylic acid ethyl ester (beta-CCE) and the corresponding methylester (beta-CCM) administered i.v. depressed segmental dorsal root potentials in spinal cats, reversed the prolongation of dorsal root potentials by phenobarbitone, and abolished the depression of a motor performance task induced by phenobarbitone in mice; beta-CCE enhanced the low-frequency facilitation of pyramidal population spikes in the hippocampus of anaesthetized rats. These effects of beta-carbolines reflect a depression of GABAergic synaptic transmission and, thus, are diametrically opposed to the enhancing action of benzodiazepine tranquilizers. The specific benzodiazepine antagonist, Ro 15-1788, while not affecting dorsal root potentials, hippocampal population spikes or phenobarbitone-induced motor performance depression, abolished the effects of beta-CCE on the three parameters and similar effects of beta-CCM on the spinal cord and motor performance. A three-state model of the benzodiazepine receptor is proposed in which benzodiazepine tranquilizers act as agonists enhancing the function of the benzodiazepine receptor as a coupling unit between GABA receptor and chloride channel, beta-carbolines act as "inverse agonists" reducing this coupling function, and Ro 15-1788 represents a competitive antagonist blocking both the enhancing effect of agonists and the depressant effect of "inverse agonists" on GABAergic synaptic transmission.

Benzodiazepine antagonist Ro 15-1788: neurological and behavioral effects.

In neurological and behavioral studies in mice, rats, dogs and squirrel monkeys, the imidazobenzodiazepinone Ro 15-1788 acted as a potent benzodiazepine antagonist. The antagonistic activity was both preventive and curative and seen at doses at which no intrinsic effects were detected. It was highly selective in that it acted against CNS effects induced by benzodiazepines but not against those produced by other depressants, such as phenobarbitone, meprobamate, ethanol, and valproate. The onset of action was rapid even after oral administration. Depending on the animal species studied, the antagonistic effects lasted from a few hours to 1 day. The acute and subacute toxicity of Ro 15-1788 was found to be very low. Benzodiazepine-like effects were not seen.