Separation of proteins by reversed-phase high-performance liquid chromatography. I. Optimizing the column.

In the process of developing a new analytical technology (the chromatophoresis process) which couples reversed-phase high-performance liquid chromatography (HPLC) to sodium dodecyl sulfate polyacrylamide gel electrophoresis in a real-time automated system, it was apparent that improvements in resolving power for the first-dimension (HPLC) separation were necessary. The present paper describes the optimization of the column for our initial work on reversed-phase HPLC separations. Polymeric (polystyrene) packings having particle diameters of 5 micron and pore diameters of 300 A were generally superior in terms of resolution, sample recovery and minimization of "ghosting".

Abel inversion using fast Fourier transforms.

A fast Fourier transform based Abel inversion technique is proposed. The method is faster than previously used techniques, potentially very accurate (even for a relatively small number of points), and capable of handling large data sets. The technique is discussed in the context of its use with 2-D digital interferogram analysis algorithms.

Uniformity of bronchoalveolar lavage in patients with pulmonary sarcoidosis.

Sarcoidosis is a granulomatous disease of unknown cause characterized by a lymphocytic alveolitis. Previous studies have shown that the inflammatory cell population of the distal lung units of patients with this disorder can be accurately assessed using bronchoalveolar lavage (BAL). The present study evaluated the uniformity of BAL between different sites of the lung in patients with sarcoidosis.

Vocal cord paresis and glottic stenosis: a late complication of poliomyelitis.

A patient with spinobulbar poliomyelitis had residual dysfunction of the ninth and tenth cranial nerves, which produced bilateral vocal cord paresis and recurrent aspiration. Critical glottic stenosis developed 28 years after the initial episode of poliomyelitis; this course appeared to be explained by fibrosis of the intrinsic laryngeal muscles and ankylosis of the right cricoarytenoid joint. Thus it appears that significant upper airway obstruction may develop as a late complication in patients with stable neurologic deficits and chronic immobility of the vocal cords.

Interaction between Candida agglutinins and antifungal agents.

Antifungal agents alter the function and morphology of Candida cell membranes and cell walls. We observed that brief (30 minute) exposure to either amphotericin B or clotrimazole inhibited the agglutination of Candida blastoconidia by murine bronchoalveolar lavage fluid. This inhibition required continuous drug presence.

Alveolar macrophage gold retention in rheumatoid arthritis.

Alveolar macrophages obtained by bronchoalveolar lavage (BAL) were evaluated by electron dispersive microanalysis (EDX) for the presence of elemental gold. EDX revealed gold in 90% (9/10) of patients with RA who were currently receiving chrysotherapy or who had discontinued chrysotherapy less than 24 months before BAL. All patients who had discontinued chrysotherapy more than 24 months before BAL (range: 3-14 years) were EDX negative (4/4), as were patients with RA who had never received gold therapy (5/5).

Phase-amplitude imaging: its application to fully automated analysis of magnetic field measurements in laser-produced plasmas.

An interferometric technique which enables simultaneous phase and amplitude imaging of optically transparent objects is discussed with respect to its application for the measurement of spontaneous toroidal magnetic fields generated in laser-produced plasmas. It is shown that this technique can replace the normal independent pair of optical systems (interferometry and shadowgraphy) by one system and use computer image processing to recover both the plasma density and magnetic field information with high accuracy. A fully automatic algorithm for the numerical analysis of the data has been developed and its performance demonstrated for the case of simulated as well as experimental data.

Candidacidal factors in murine bronchoalveolar lavage fluid.

Respiratory secretions provide an efficient method for protecting the large surface area of the lower respiratory tract. To determine whether lung secretions contribute to antifungal defenses, we tested bronchoalveolar lavage fluid for fungicidal activity. Candida albicans (blastoconidia) was incubated in unconcentrated cell-free lavage fluid from Swiss Webster mice and then cultured quantitatively to measure residual viability.

Coded aperture imaging: a Fourier space analysis.

Coded aperture imaging is analyzed in Fourier space and the conditions for obtaining artifact-free 3-D images are obtained. It is deduced that an infinite square array of coding apertures will obey these conditions. A finite array is considered and it is shown that, after a certain coordinate transformation has been performed, the finite aperture acts to bandlimit the spatial frequencies in the image.

Effect of Candida morphology on amphotericin B susceptibility.

We showed that brief exposures to amphotericin B (AmB) inhibited the induction of new Candida germ tubes and the lengthening of partially induced germ tubes. Blastoconidia with germ tubes were more susceptible to AmB killing, and this varied directly with the induction period and the AmB exposure period. AmB did not preferentially affect germ tube adherence to fibrin matrices.

Comparison of automated pre-column and post-column analysis of amino acid oligomers.

It has been shown that various amino acids will polymerize under plausible prebiotic conditions on mineral surfaces, such as clays and soluble salts, to form varying amounts of oligomers (n = 2-6). The investigations of these surface reactions required a quantitative method for the separation and detection of these amino acid oligomers at the picomole level in the presence of nanomole levels of the parent amino acid. In initial high-performance liquid chromatography (HPLC) studies using a classical postcolumn o-phthalaldehyde (OPA) derivatization ion-exchange HPLC procedure with fluorescence detection, problems encountered included lengthy analysis time, inadequate separation and large relative differences in sensitivity for the separated species, expressed as a variable fluorescent yield, which contributed to poor quantitation.

Effect of alkylating agents on the clearance of Staphylococcus aureus from murine lungs.

Alkylating agents have several effects on cellular host defense responses which could increase either the frequency or the severity of pulmonary infections. In addition, some of these agents directly injure lung parenchyma and could have effects on intrapulmonary killing processes independent of any effect on phagocyte number and function. We have used a murine model for staphylococcal clearance to evaluate the effect of cyclophosphamide and mechlorethamine on intrinsic lung defenses.

Effects of sublethal concentrations of amphotericin B on Candida albicans.

Because sublethal concentrations of antibodies can have important effects on bacteria and may aid host defenses, even in the absence of direct microbial killing, the effect of brief sublethal exposures to amphotericin B on Candida albicans blastoconidia was evaluated. Amphotericin B (0.01-1.

Interferon and growth factor activity for human lung fibroblasts. Release from bronchoalveolar cells from patients with active sarcoidosis.

Pulmonary sarcoidosis is a granulomatous disorder of unknown etiology characterized by both an active cellular immune process and interstitial fibrosis. It has recently been demonstrated that gamma-interferon (the type of interferon associated with an active cellular immune process) is also a potent growth factor for human lung fibroblasts. To evaluate the hypothesis that there is an association between the release of immune interferon and the release of growth factor activity for fibroblasts, bronchoalveolar cells from patients with sarcoidosis were studied for the spontaneous release of both immune interferon and growth factor activity for fibroblasts.

Site of airflow obstruction during early and late phase asthmatic responses to allergen bronchoprovocation.

Chronic asthma and late asthmatic responses (LAR) are associated with local inflammation which might be expected to produce airflow obstruction in small airways and to increase nonspecific airway reactivity. In contrast, early asthmatic responses (EAR) are primarily bronchospastic and probably involve more central airways. We challenged 17 nonsmoking, mildly asthmatic atopic subjects with allergen bronchoprovocation and measured changes in spirometry (FEV1) over the next 24 hours.

Inhibitory effects of chlorpromazine on Candida species.

Chlorpromazine was tested for antifungal activity by using Candida albicans and standard assays. The MIC of chlorpromazine was 35 micrograms/ml; the minimal fungicidal concentration was also 35 micrograms/ml. The minimal effective concentration was 2.

Stimulated human alveolar macrophages secrete interferon.

Alveolar macrophages were isolated by bronchoalveolar lavage from normal subjects to determine whether these cells can be activated to produce interferon. Macrophages were incubated for 24 h, and the supernatants were assayed for interferon using a plaque reduction assay (vesicular stomatitis virus and human amnion cells). The macrophages did not spontaneously release detectable amounts of interferon, but macrophages stimulated with either mitogens or classic inducers did release antiviral activity (titer range, 32 to 962 units/ml).