AANA Journal course: update for nurse anesthetists--anesthesia for the ruptured globe.

Anesthesia for the patient with a perforated globe can be complicated. Cognizance of the anatomy and physiology of the eye, including maintenance of intraocular pressure, is essential for the development of an anesthetic plan. Since the induction phase of anesthesia is the most critical period during which intraocular pressure is affected, understanding the pharmacology of the various anesthetic agents and their effects on the eye is important. To avoid increasing intraocular pressure, a smooth, atraumatic induction is desired. However, methods to achieve this end may place the patient at risk for aspiration. Various techniques that attempt to accomplish this goal are described, including the use of narcotics, lidocaine, nitroglycerin, alpha (alpha 2) agonism, beta (beta) adrenergic and calcium channel blockades, plus the laryngeal mask airway.

A chemotaxis cluster from Agrobacterium tumefaciens.

We report the DNA sequence of a 9.6-kb region of the Agrobacterium tumefaciens chromosome containing a putative 8-kb chemotaxis operon. The putative operon begins with orf1, whose predicted protein product shows strong sequence identity to methyl-accepting chemotaxis proteins (MCPs), followed by orf2, cheY1, cheA, cheR, cheB, cheY2, orf9, orf10. All of the identified homologues show a high degree of sequence conservation with their counterparts in the che operons from Sinorhizobium meliloti and Rhodobacter sphaeroides, and are arranged in a similar order. Mutations in orf1 and cheA result in impaired chemotaxis, whereas deletion of orf10, appears to have no effect on chemotaxis or motility. Although the putative operon does not contain a cheW homologue, heterologous probing and PCR using consensus primers indicates that cheW maps elsewhere in the Agrobacterium genome.

Music training causes long-term enhancement of preschool children's spatial-temporal reasoning.

Predictions from a structured cortical model led us to test the hypothesis that music training enhances young children's spatial-temporal reasoning. Seventy-eight preschool children participated in this study. Thirty-four children received private piano keyboard lessons, 20 children received private computer lessons, and 24 children provided other controls. Four standard, age-calibrated, spatial reasoning tests were given before and after training; one test assessed spatial-temporal reasoning and three tests assessed spatial recognition. Significant improvement on the spatial-temporal test was found for the keyboard group only. No group improved significantly on the spatial recognition tests. The magnitude of the spatial-temporal improvement from keyboard training was greater than one standard deviation of the standardized test and lasted at least one day, a duration traditionally classified as long term. This represents an increase in time by a factor of over 100 compared to a previous study in which listening to a Mozart piano sonata primed spatial-temporal reasoning in college students. This suggests that music training produces long-term modifications in underlying neural circuitry in regions not primarily concerned with music and might be investigated using EEG. We propose that an improvement of the magnitude reported may enhance the learning of standard curricula, such as mathematics and science, that draw heavily upon spatial-temporal reasoning.

Monoclonal infection involving Mycobacterium avium presenting with three distinct colony morphotypes.

Recent reports indicate that polyclonal infections may play an important role in multiple drug resistance in Mycobacterium avium infections. We report here on the isolation of a single M. avium strain that appeared to have smooth colony morphology upon initial isolation on a Lowenstein-Jensen slant. Primary subculture onto Middlebrook 7H10, however, revealed three distinct morphotypes representing smooth opaque (SmO), smooth transparent (SmT), and rough (Rg) colony morphologies. All three morphotypes were identified as M. avium by standard biochemical procedures, Genprobe analysis, and mycolic acid patterns. Subsequent restriction fragment length polymorphism analysis, using SalI- and PvuII-digested genomic DNA, revealed identical patterns for hybridization with the IS1245 probe. Thin-layer chromatographic analysis of lipids from the three morphotypes revealed that only the SmT morphotype possessed what appeared to be lipid components similar to, but unlike, previously described serovar-specific glycopeptidolipid antigens. Further analysis of internally radiolabeled deacylated lipids from the SmT morphotype, by high-performance liquid chromatography and thin-layer chromatography, disclosed that some of these components can be internally radiolabeled with [14C] phenylalanine and [14C]mannose. These results suggest that these components are structurally similar to previously described glycopeptidolipid antigens. This is apparently the first report of a monoclonal infection involving a single strain of M. avium presenting with all three colony morphotypes, SmO, SmT, and Rg.

A unique phenylalanine-containing lipopeptide isolated from a rough-colony variant of Mycobacterium avium.

Previous investigations have suggested that the biosynthesis of the Mycobacterium avium serovar-specific glycopeptidolipid antigens involves initial steps that include the participation of lipopeptides. The prevailing assumption is that subsequent glycosylation of those lipopeptides results in the fully glycosylated form of the glycopeptidolipid components. In an effort to identify potential precursors in the biosynthetic pathway of glycopeptidolipid components, we have identified a unique lipopeptide from an M. avium rough variant (MAC702) that was isolated from a patient suffering from a chronic M. avium lung infection. Upon examination it was revealed that although the total lipid extract from MAC702 lacked serovar-specific glycopeptidolipid antigens, it did contain a unique lipopeptide, possessing some amino acids identical to those found in the serovar-specific glycopeptidolipid antigens. Initial examination of acid-hydrolyzed samples of the lipopeptide (lipopeptide-I) revealed the presence of phenylalanine, alanine, and isoleucine, but no carbohydrate. Subsequent mass spectrometric and 1H-NMR and 1H-13C-NMR correlation spectroscopy analysis confirmed the initial results and also revealed the presence of N-methylisoleucine. The following structure for lipopeptide-I was proposed: fatty acyl (C19 or C17)-Phe-N-methyl-Ile-Ile-Phe-Ala-Ile-Ala-Phe. Lipopeptide-I is unlike any heretofore identified compound, however, it does have similar features to lipopeptides previously reported in mycobacteria and fungi. Although its structure does not verify that it is a direct precursor in glycopeptidolipid biosynthesis, the presence of certain components in lipopeptide-I indicate that it may share at least some pathways associated with the biosynthesis of the M. avium serovar-specific glycopeptidolipids.

Use of Mono Mac 6 human monocytic cell line and J774 murine macrophage cell line in parallel antimycobacterial drug studies.

The Mono Mac 6 (MM6) human monocytic cell line was evaluated with the established J774 murine macrophage cell line to ascertain its effectiveness in determining the intracellular activities of antimycobacterial drugs. Cells were infected with Mycobacterium tuberculosis H37Ra and treated with drug concentrations corresponding to the MICs, as well as to threefold higher than and threefold less than the MICs. Changes in CFU were compared after 7 days to determine significant differences between treated and nontreated groups. The results suggest that MM6 will make a useful model for testing the intracellular activities of antituberculosis drugs.

Immunomodulatory spectrum of lipids associated with Mycobacterium avium serovar 8.

Lipid fractions obtained from Mycobacterium avium serovar 8 were assessed for the ability to affect various immune functions of human peripheral blood mononuclear cells (PBM). Lipids included a total lipid fraction and fractions eluted from silicic acid column separation of that total lipid fraction, using chloroform and chloroform-methanol combinations. Lipid fractions were assayed for total carbohydrate and total 6-deoxyhexose content and were assessed for the ability to influence human macrophage function and the capacity to induce secretion of prostaglandin E2 (PGE2) and tumor necrosis factor alpha in PBM. The total lipid and serovar-specific glycopeptidolipid (GPL) fractions both induced significant levels of tumor necrosis factor alpha, as well as PGE2, in PBM exposed to a sublethal concentration of 100 micrograms lipid per 2 x 10(6) cells. In addition, the same concentrations of the 5 to 7% and GPL fractions induced significant levels of leukotriene B4 in PBM. Comparison of carbohydrate and 6-deoxyhexose contents of each fraction suggested a relationship to carbohydrate content and ability of fractions to induce immune modulator secretion. Analysis of GPL fractions from M. avium serovars 4 and 20 revealed that those GPL lacked the ability to induce PGE2. These results are explained by considering the difference in the carbohydrate residues of the oligosaccharide moieties.

Potential drug targets for Mycobacterium avium defined by radiometric drug-inhibitor combination techniques.

Previously established radiometric techniques were used to assess the effectiveness of combined antimicrobial drug-inhibitory drug (drug-inhibitor) treatment on two clinical isolates of the Mycobacterium avium complex representing three colony variants: smooth opaque (dome) (SmO), smooth transparent (SmT), and rough (Rg). All variants were identified as members of the M. avium complex; however, only the SmT colony type of strain 373 possessed characteristic serovar-specific glycopeptidolipid (GPL) antigens. MICs, determined radiometrically, of drugs with the potential to inhibit the biosynthesis of GPL antigens or other cell envelope constituents were similar for all strains. These drugs included cerulenin, N-carbamyl-DL-phenylalanine, N-carbamyl-L-isoleucine, trans-cinnamic acid, ethambutol, 1-fluoro-1-deoxy-beta-D-glucose, 2-deoxy-D-glucose, and m-fluoro-phenylalanine. The MICs of the antimicrobial drugs amikacin, sparfloxacin, and clarithromycin varied, but overall the MICs for the SmO variant were the lowest. Radiometric assessment of drug-inhibitor combinations by using established x/y determinations revealed enhanced activity when either ethambutol or cerulenin were used in combination with all antimicrobial agents for all variants except the Rg variant of strain 424, for which ethambutol was not effective. Enhanced activity with amino acid analogs was observed with the Rg colony variants of strains 373 and 424. Two potential sites for drug targeting were identified: fatty acid synthesis, for all strains assayed, and peptide biosynthesis, particularly for Rg colony variants that possess previously identified phenylalanine-containing lipopeptides as potential targets for future drug development.

Immunomodulation of human peripheral blood mononuclear cell functions by defined lipid fractions of Mycobacterium avium.

Mycobacterial fractions, some of which are associated with the cell envelope of Mycobacterium avium serovar 4, were assessed for their ability to affect various immunological functions of human peripheral blood mononuclear cells (PBM). Treatment of PBM with a total lipid fraction derived from M. avium serovar 4 resulted in a significant suppression of lymphoproliferative responsiveness to phytohemagglutinin stimulation at concentrations not affecting cell viability. Although a similar suppression was not observed when PBM were treated with purified serovar 4-specific glycopeptidolipids (GPL), treatment with the beta-lipid fragment derived from the GPL did result in a significant suppression of phytohemagglutinin responsiveness. Further studies revealed that the total lipid fraction and the beta-lipid fragment were effective at significantly reducing the ability of human macrophages to restrict the intracellular growth of mycobacteria and at stimulating PBM to secrete prostaglandin E2. These same effects were not observed when purified GPL or the reduced oligosaccharide fragment of the GPL was used. Other studies revealed that the total lipid and purified GPL fractions were effective at stimulating tumor necrosis factor alpha release from human PBM, whereas the beta-lipid fragment was not. These results indicate that mycobacterial lipids have various immunomodulatory capabilities, depending upon their chemical nature and ability to interact with certain host cells.

Durations of extended mental rehearsals are remarkably reproducible in higher level human performances.

It has been extremely difficult to quantify temporal aspects of higher level human brain function. We have found that mental rehearsals of musical performance of several minutes duration provide such a measure in that they can be highly reproducible, varying to less than 1%. These remarkable results pose fundamental neurophysiological problems. It is necessary to understand the underlying neuronal bases for this accuracy in the spatial-temporal activity of billions of neurons over minutes without sensory input. Further, they present a powerful constraint on neuronal models of brain function. Such highly reproducible (in duration) mental rehearsals might be used in conjunction with multielectrode EEG recordings to look for reproducible spatial-temporal patterns. Further, we suggest that our results may provide an extremely useful behavioural correlate for high level performance.

Scientific results from the Cosmic Background Explorer (COBE).

The National Aeronautics and Space Administration (NASA) has flown the COBE satellite to observe the Big Bang and the subsequent formation of galaxies and large-scale structure. Data from the Far-Infrared Absolute Spectrophotometer (FIRAS) show that the spectrum of the cosmic microwave background is that of a black body of temperature T = 2.73 +/- 0.06 K, with no deviation from a black-body spectrum greater than 0.25% of the peak brightness. The data from the Differential Microwave Radiometers (DMR) show statistically significant cosmic microwave background anisotropy, consistent with a scale-invariant primordial density fluctuation spectrum. Measurements from the Diffuse Infrared Background Experiment (DIRBE) provide new conservative upper limits to the cosmic infrared background. Extensive modeling of solar system and galactic infrared foregrounds is required for further improvement in the cosmic infrared background limits.

Activities of fluoroquinolone, macrolide, and aminoglycoside drugs combined with inhibitors of glycosylation and fatty acid and peptide biosynthesis against Mycobacterium avium.

Smooth- and rough-colony variants of Mycobacterium avium serovar 4 were treated with three classes of drugs. The drugs were chosen for their potential inhibitory effects on the biosynthesis of the cell envelope-associated serovar-specific glycopeptidolipid antigens. Growth was monitored radiometrically with a BACTEC 460-TB instrument, and MICs were determined for each drug. Both variants were then treated with inhibitory drugs in combination with antimicrobial agents that have demonstrated effectiveness against M. avium. No growth inhibition was observed with 6-fluoro-6-deoxy-D-glucose or avidin. Inhibitors of glycosylation, i.e., 2-deoxy-D-glucose, bacitracin, and ethambutol, were inhibitory to smooth- and rough-colony variants, whereas drugs that inhibit peptide synthesis, i.e., N-carbamyl-L-isoleucine and m-fluoro-phenylalanine, were more inhibitory for the rough-colony variant. Cerulenin, which affects fatty acid synthesis, was inhibitory for both variants, but it appeared to be more effective at inhibiting the growth of the smooth-colony variant at equivalent concentrations. Generally, when inhibitors of glycosylation were used with sparfloxacin and amikacin, a synergistic effect was observed for only the smooth variant. When drugs that affect peptide synthesis were used in combination with amikacin, a synergistic effect was observed for the rough variant, and when cerulenin was used in combination with sparfloxacin or amikacin, a synergistic effect was observed for both variants. Lipid analysis revealed that although the rough variant lacks the serovar-specific glycopeptidolipid antigens, it does possess a group of phenylalanine-isoleucine-containing lipopeptides that may explain its different susceptibility patterns to m-fluoro-phenylalanine and N-carbamyl-L-isoleucine. The significance of these results is discussed with reference to various components in the cell envelope and their importance in cell wall permeability.

Mycobacterium avium rough-to-smooth colony conversion resulting from growth in Tween 80 without presence of type-specific glycopeptidolipid antigens.

Growth of a Mycobacterium avium complex serotype 20 rough-colony variant in 1.0% Tween 80 resulted in a smooth-colony morphological conversion that was reversible upon removal of Tween and was not associated with the presence of serotype-specific glycopeptidolipid antigens. Electron microscopic examination suggested the role of an outer layer in the Tween-related morphological modification.

In vivo binding and clearance of circulating antigen by bispecific heteropolymer-mediated binding to primate erythrocyte complement receptor.

We have used the avidin/biotin system to construct soluble, cross-linked bispecific heteropolymers containing mAb to both the primate E C receptor and the DNP group. These heteropolymers facilitate in vitro binding of DNP-bovine gamma-globulin (DNP-BGG) to both human and squirrel monkey E. Intravenous injection in squirrel monkeys of DNP-BGG followed by heteropolymer leads to E binding and clearance from the circulation of a significant fraction of both heteropolymer and DNP-BGG, without lysis or clearance of the E. This methodology may potentially be used to treat a variety of infectious diseases and other syndromes associated with blood-borne pathogens.

Modified lymphocyte response to mitogens induced by the lipopeptide fragment derived from Mycobacterium avium serovar-specific glycopeptidolipids.

The beta-elimination procedure was used to obtain two major fragments of Mycobacterium avium glycopeptidolipid antigens. The lipopeptide fragment, not the oligosaccharide, diminished the mitogen-induced blastogenic response of spleen cells at concentrations lower than those which affected viability. Electron microscopy revealed an internalization of lipopeptide and disruption of intracellular organization.

Cell-free system responsible for internal radiolabeling of glycopeptidolipids of the Mycobacterium avium complex.

Internal radiolabeling of serotype-specific glycopeptidolipids with [14C]mannose was accomplished with a cell-free system derived from serotype 20 of the Mycobacterium avium complex. Similar radiolabeling was not apparent with a cell-free system derived from the rough colony variant, previously shown to be devoid of glycopeptidolipids. Although a comparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein analysis of the parent and rough variant strains revealed a close similarity, there were some proteins unique to the parent strain.

Complement-opsonized IgG antibody/dsDNA immune complexes bind to CR1 clusters on isolated human erythrocytes.

We used fluorescence microscopy, quantitative FACS analyses, and radioimmunoassays to examine the distribution of complement (C3b)-opsonized antibody/dsDNA immune complexes (IC) bound to normal human erythrocytes (RBCs) via immune adherence (IA). IC were detected with fluorescent anti-human IgG, and the RBC IA-receptor (CR1) was revealed with monoclonal antibodies (Mabs) to CR1 and fluorescent anti-mouse IgG. Under saturating conditions RBCs exhibit a large heterogeneity in binding; a significant fraction binds no IC. The positions of bound IC coincide with CR1 clusters on RBCs, as predicted by several investigators. FACS experiments indicate an excellent correlation between CR1 number and IC binding within an RBC population. The number of CR1 clusters able to bind IC is proportional to the number of CR1 per RBC. However, IC (fluorescent spots) detected per RBC (on average less than 10) are less than the average number of IC bound (20-30). This suggests that each fluorescent spot represents a small number of aggregated IC bound to a CR1 cluster. These "patches" of aggregated complexes may facilitate transfer of RBC-bound IC to cells of the mononuclear phagocytic system. Finally, not all the CR1 clusters bind IC, suggesting that proper geometric alignment of multiple C3b per IC with several CR1 in the cluster is required for IA.

Inhibition of glycopeptidolipid synthesis resulting from treatment of Mycobacterium avium with 2-deoxy-D-glucose.

Exponentially growing cultures of Mycobacterium avium complex serovar 4 were treated with 2-deoxy-D-glucose (2-DDG) and incubated with radiolabelled components which incorporate into the serovar-specific glycopeptidolipids (GPL) associated with the L1 layer. Following treatment with the drug, radiolabelled lipids were extracted from the mycobacteria and examined by thin-layer chromatography, high performance liquid chromatography and autoradiography to determine the percent distribution of radioactivity in the GPL and other related lipids. Treatment of serovar 4 with 2-DDG resulted in a dose-dependent inhibition of GPL biosynthesis, as judged by a reduction in the incorporation of radiolabelled phenylalanine, mannose and methionine into the GPL. In addition, a concomitant accumulation of at least two phenylalanine-containing lipopeptides was observed in cells treated with 2-DDG. Cultivation of serovar 4 in the presence of 2-deoxy-D-1,2-(3)H-glucose did not result in internal radiolabelling of the GPL, indicating that 2-DDG was not being incorporated into the GPL as an analogue of mannose, but rather was acting as a metabolic inhibitor of GPL biosynthesis.