Cellular localizations of AMPA glutamate receptors within the basal forebrain magnocellular complex of rat and monkey

The cellular distributions of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors within the rodent and nonhuman primate basal forebrain magnocellular complex (BFMC) were demonstrated immunocytochemically using anti-peptide antibodies that recognize glutamate receptor (GluR) subunit proteins (i.e., GluR1, GluR4, and a conserved region of GluR2, GluR3, and GluR4c). In both species, many large GluR1-positive neuronal perikarya and aspiny dendrites are present within the medial septal nucleus, the nucleus of the diagonal band of Broca, and the nucleus basalis of Meynert. In this population of neurons in rat and monkey, GluR2/3/4c and GluR4 immunoreactivities are less abundant than GluR1 immunoreactivity. In rat, GluR1 does not colocalize with ChAT, but, within many neurons, GluR1 does colocalize with GABA, glutamic acid decarboxylase (GAD), and parvalbumin immunoreactivities. GluR1- and GABA/GAD-positive neurons intermingle extensively with ChAT-positive neurons. In monkey, however, most GluR1-immunoreactive neurons express ChAT and calbindin-D28 immunoreactivities. The results reveal that noncholinergic GABAergic neurons, within the BFMC of rat, express AMPA receptors, whereas cholinergic neurons in the BFMC of monkey express AMPA receptors. Thus, the cellular localizations of the AMPA subtype of GluR are different within the BFMC of rat and monkey, suggesting that excitatory synaptic regulation of distinct subsets of BFMC neurons may differ among species. We conclude that, in the rodent, BFMC GABAergic neurons receive glutamatergic inputs, whereas cholinergic neurons either do not receive glutamatergic synapses or utilize GluR subtypes other than AMPA receptors. In contrast, in primate, basal forebrain cholinergic neurons are innervated directly by glutamatergic afferents and utilize AMPA receptors.     

Use of a tapered fluidized bed as a continuous bioreactor

Reactor systems based on tapered fluidized beds are being developed for aqueous bioprocesses in which adhering microorganisms or immobilized active biological fractions are used. The use of a fluidized bed prevents biomass buildup, accommodates particulates in the feed stream, is compatible with gas sparging, and allows easy removal or addition of the active materials. The tapered reactor tends to stabilize the fluidized bed, thus allowing a much wider range of operating conditions. Preliminary experimental results and an empirical mathematical model of the tapered bed indicate that bed stability is associated with a decreasing velocity and void-fraction profile up the bed and the pressure drop across the bed decreases with increasing flow rates. The tapered fluidized bed bioreactor is being evaluated for use in the enzymatic production of hydrogen, microbiological denitrification, and microbiological degradation of coal conversion aqueous waste streams. The enzyme catalyzed conversion of lactose to glucose and galactose was used in the evaluation of the reactor concept.     

Regional hip blockade. A simplified technique for the relief of intractable osteoarthritic pain

A regional anaesthetic technique is described using a blind anatomical approach to the obturator nerve and nerve to quadratus femoris, as a means of alleviating the disabling pain of chronic osteoarthritis of the hip, when arthroplasty is not available.     

Causes of initial visual loss following blunt ocular injuries: an experimental study

We have investigated in anaesthetised pigs the electrical responses of the retina to light after blunt injuries to the eye. Retinograms, cortical visual evoked responses and intraocular pressures were recorded before and after non-perforating mechanical trauma of sufficient severity to produce traumatic retinal opacification. After injury, applanation pressures rose up to 34 mm. Hg above normal, reaching peak values 4 to 12 minutes after trauma and returning to pre-trauma levels after about 45 minutes. These pressure changes were not responsible for the immediate abolition of the B wave of the retinogram or of the cortical visual evoked responses which occurred after trauma, but increases of intraocular pressure modified the recovery pattern of the visual evoked response.     

Oncostatin M, but not interleukin-6 or leukemia inhibitory factor, stimulates expression of alpha1-proteinase inhibitor in A549 human alveolar epithelial cells

Alpha-1 proteinase inhibitor (A1-Pi) is the main serine proteinase inhibitor found in human plasma and is a potent elastase inhibitor in various tissues, including lung. A1-Pi is expressed and induced in liver during inflammatory responses but can also be produced by epithelial cells. Since hepatocyte A1-Pi production is stimulated by interleukin-6 (IL-6) and other gp130-cytokines, such as leukemia inhibitory factor (LIF) and oncostatin M (OM), we investigated the role of these cytokines in regulating A1-Pi in lung epithelial cells. We show that OM, a monocyte and T cell product, can specifically and potently induce A1-Pi production in lung-derived A549 alveolar (epithelial) cells, as well as in liver-derived HepG2 cells. Both A1-Pi protein (as detected by ELISA and Western blots) and mRNA levels were enhanced 20-fold to 30-fold in A549 cells. OM was also able to stimulate the expression of tissue inhibitor of metalloproteinase-1 in these cells. Interestingly, other members of the IL-6 family (IL-6 and LIF) had little or no effect on A549 cells, and proinflammatory cytokines, such as IL-1 beta and tumor necrosis factor-alpha (TNF-alpha) also had no stimulatory effect on A1-Pi synthesis in A549 cells. Costimulation with IL-1 beta resulted in a decrease in A1-Pi production from OM-stimulated A549 cells. However, IL-6 production was synergistically enhanced. OM was also able to stimulate A1-Pi production from a bronchial epithelial primary cell line, whereas an intestinal epithelial cell line HT29 responded to IL-6 but not OM. These results suggest that lung levels A1-Pi could be derived not only from liver and inflammatory cells but also from epithelial cells, which can be upregulated on stimulation by OM. This may have implications for regulation of local activity of human neutrophil elastase (HNE) in such diseases as emphysema and cystic fibrosis.     

Effects of gastric mucosal blood flow (GMBF) on the role of adaptive cytoprotection of rat gastric mucosa

It has been reported that ingestion of an ammonium-containing diet produces hyperammonemia without encephalopathy, thus permitting the study of the specific effects of ammonia toxicity. The present study investigated the rat cerebral somatostatinergic system using this experimental model of hyperammonemia. Wistar rats were fed a high ammonia diet prepared by mixing a standard diet with ammonium acetate (20% w/w); in addition, 5 mM of ammonium acetate was added to their water supply. Control rats were fed with a standard diet. The animals were sacrificed at 3, 7 and 15 days of ammonia ingestion. Ammonia levels in blood had increased approximately 3-fold at 7 days of ammonia ingestion. These changes were associated with a significant decrease in the specific binding of somatostatin (SS) to putative receptors sites in the frontoparietal cortex and hippocampus at 7 and 15 days after starting the high ammonia diet. Scatchard analysis shows that the decrease in SS binding resulted from a decrease in the number of available SS receptors rather than a change in receptor affinity. No changes in the somatostatin-like immunoreactivity content (SSLI) were detected in either brain area at the three study times. These results suggest that hyperammonemia alone can affect the rat brain somatostatinergic system. However, the animal model of hyperammonemia used here is insufficient to produce encephalopathy despite the significant increase in serum ammonia.     

Use of the recombinant 38-kDa antigen of Mycobacterium tuberculosis as an immunogen for specific antisera production

The Mycobacterium tuberculosis 38-kDa protein antigen is one of the secreted immunodominant antigens showing high immunogenicity at B-cell and T-cell levels. Although monoclonal antibodies to this antigen have been produced, specific polyclonal antisera is required for standardization of specific immunodiagnostic assays. This protein has been overexpressed and purified from recombinant Escherichia coli using an inducible vector system. During each stage of expression and purification, the recombinant protein was used to immunize mice and rabbits by several methods: 1) as overexpressed protein present as inclusion bodies in recombinant E. coli; 2) embedded in a polyacrylamide gel; 3) fixed to a solid-phase nitrocellulose membrane and 4) emulsified with an adjuvant. All strategies yielded specific antisera as determined by enzyme-linked immunosorbent assay (ELISA) and immunoblot analyses. The results obtained, both quantitative (ELISA) and qualitative (immunoblot) demonstrate that the purified recombinant antigen retains its antigenicity and immunogenicity throughout the various steps in the process of expression and purification and serves as a potent antigen for production of specific antisera to be used in immunoassays.     

Development of antibiotic-resistant strains for the enumeration of foodborne pathogenic bacteria in stored foods

Strains of Aeromonas spp., Salmonella enteritidis phage type 4, Salmonella typhimurium, verotoxigenic Escherichia coli O157:H7 (VTEC) and Yersinia enterocolitica resistant to streptomycin, nalidixic acid and a combination of both antibiotics were selected. When compared with the parent strains, most of the antibiotic-resistant strains had slightly slower growth rates at their optimum incubation temperature but the difference was reduced progressively when the temperature was lowered. Some antibiotic-resistant strains had considerably slower growth rates in the presence of the relevant antibiotic and these were not used further. Several agar and impedance media with added streptomycin and nalidixic acid were assessed for the enumeration of the antibiotic-resistant strains in artificially contaminated stored foods. Differential/selective media were required to enumerate low numbers of antibiotic-resistant strains in certain foods. The following agar and impedance media were selected: Aeromonas Agar (Ryan) for Aeromonas spp., Xylose Lysine Agar and Lysine Iron Cysteine Neutral Red Medium for Salmonella, Eosin Methylene Blue Agar and Coliform Medium for VTEC, and Yersinia Selective Agar without selective agents for Yersinia enterocolitica. The agar and impedance media have been used successfully to enumerate antibiotic-resistant strains inoculated into foods and stored at different temperatures.     

Global increases in seizure susceptibility in mice lacking 5-HT2C receptors: a behavioral analysis

OBJECTIVE: Vasodilation by beta-adrenergic receptors of smooth muscle cells appears to be impaired early after the onset of hypercholesteremia. The aim of this study was to analyze the modulation of beta-adrenergic receptor density and adenylyl cyclase activity in the presence of moderately elevated concentrations of LDL. The effects of beta 1- and beta 2-adrenergic receptor antagonists on LDL-induced receptor changes were studied. METHODS AND RESULTS: Media explants of porcine coronary arteries were incubated with moderately elevated LDL concentrations (0.7-3.9 mmol/l). The density of beta-adrenergic receptors was determined in plasma membranes using the radioligand [125I]iodocyanopinodolol. LDL (3.9 mmol/l) resulted in a decrease of beta-adrenergic receptor density (control 137 +/- 5 vs. 89 +/- 7 fmol/mg protein, P < 0.01). After removal of LDL and cultivation for an additional 3 days beta-adrenergic receptors increased to 129 +/- 5 fmol/mg. In the presence of the beta 1- or beta 2-adrenergic receptor antagonists the LDL-mediated decrease was inhibited. Addition of metoprolol after 3 days of LDL incubation caused a restoration of receptor density. The basal, isoproterenol- and forskolin-stimulated adenylyl cyclase activities were increased after LDL incubation by 180, 110 or 80%, respectively. CONCLUSION: Moderately elevated LDL levels decreased beta-adrenergic receptor density while adenylyl cyclase activity was simultaneously increased. beta 1- or beta 2-adrenergic receptor antagonists prevented this receptor decrease and might preserve the beta-adrenergic receptor density in the presence of moderately elevated LDL levels.