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Biomedical subjects

Steven J Mentzer

Publications and source records attributed to Steven J Mentzer.

25 records · Page 2Linked to original sources

Biochemical changes in the efferent lymph plasma after oxazolone stimulation.

The lymph node is a specialized microenvironment for the regulation of immune responses. The reticular network of the lymph node provides a structure that facilitates not only intercellular interactions, but the intranodal flow of lymph fluid. To assess biochemical changes in the nodal lymph plasma after antigen stimulation, prescapular lymph nodes in sheep were stimulated with the epicutaneous antigen oxazolone. The efferent lymph from both antigen-stimulated and contralateral control prescapular lymph nodes was continuously monitored for more than 120 h. The oxazolone-stimulated lymph plasma was associated with a selective increase in cholesterol content during the 'recruitment' phase of lymph node enlargement. The peak in cholesterol was followed closely by a significant increase in lymph plasma LDH concentration. In contrast, there was no significant difference between oxazolone-stimulated and control lymph plasma in the concentration of triglycerides, albumin, alkaline phosphatase, and alanine transferase. These selective biochemical changes in the efferent lymph appeared to reflect the dynamics of lymphocyte activation within the lymph node as well as provide a practical measure of the lymph node response to antigen stimulation.

Animals↗

Stimulation of regional lymphatic and blood flow by epicutaneous oxazolone.

The application of the epicutaneous antigen oxazolone results in persistent induration and erythema; however, the relative changes in lymph and blood flow in the inflammatory skin are largely unknown. To define the contribution of lymph and blood flow to the clinical appearance of cutaneous inflammation, we studied the sheep ear after the application of oxazolone. As a model for the study of these changes, the sheep ear had several experimental advantages: 1) a simplified superficial vascular network, 2) defined lymphatic drainage, and 3) an avascular and alymphatic cartilaginous barrier. Lymph flow was continuously monitored by cannulation of the prescapular efferent lymph duct. Blood flow, as reflected by cutaneous erythema, was noninvasively measured by use of a visible-spectrum spectrophotometer. The application of the epicutaneous oxazolone resulted in increased ear thickness for >7 days. The lymph flow from the oxazolone-stimulated ear peaked between 24 and 48 h after oxazolone stimulation. Spectrophotometric evaluation indicated that the cutaneous erythema peaked 72-96 h after application of oxazolone. Corrosion casting and scanning electron microscopy of the microcirculation at 96 h after antigen stimulation demonstrated significant dilatation of the superficial vascular network. These results suggest a biphasic response to oxazolone stimulation: 1) an early increase in vascular permeability associated with increased lymph flow and 2) a subsequent increase in relative blood flow associated with a dilated inflammatory microcirculation.

Adjuvants, Immunologic↗

Using a neural network with flow cytometry histograms to recognize cell surface protein binding patterns.

Flow cytometric systems are being used increasingly in all branches of biological science including medicine. To develop analytic tools for identifying unknown molecules such as the antibodies that recognize different structure in the identical antigens, we explored use of a neural network in flow cytometry data comparison. Peak locations were extracted from flow cytometry histograms and we used the Marquardt backpropagation neural networks to recognize identical or similar binding patterns between antibodies and antigens based on the peak locations. The neural network showed 93.8% to 99.6% correct classification rates for identical or similar molecules. This suggests that the neural network technique can be useful in flow cytometry histogram data analysis.

Antibodies, Monoclonal↗

Matching of flow-cytometry histograms using information theory in feature space.

Flow cytometry is a widely available technique for analyzing cell-surface protein expression. Data obtained from flow cytometry is frequently used to produce fluorescence intensity histograms. Comparison of histograms can be useful in the identification of unknown molecules and in the analysis of protein expression. In this study, we examined the combination of a new smoothing technique called SiZer with information theory to measure the difference between cytometry histograms. SiZer provides cross-bandwidth smoothing and allowed analysis in feature space. The new methods were tested on a panel of monoclonal antibodies raised against proteins expressed on peripheral blood lymphocytes and compared with previous methods. The findings suggest that comparing information content of histograms in feature space is effective and efficient for identifying antibodies with similar cell-surface binding patterns.

Animals↗

Melittin-induced membrane permeability: a nonosmotic mechanism of cell death.

Derived from honeybees, melittin is a 26-amino acid, alpha-helical, membrane-attack protein that efficiently kills mammalian cells. To investigate the contribution of colloid-osmotic effects to the mechanism of cell death, we studied the effect of melittin on lymphocyte membrane permeability and cell volumes. Melittin concentrations of 0.5 to 2.0 microM induced release of membrane permeability markers without total disruption of the cell membrane. At these melittin concentrations, electrical-impedance cytometry demonstrated melittin-induced changes in red blood cell volumes (P<0.01), but no change in lymphocyte cell volumes (P>0.05). Streaming video microscopy, obtaining images of melittin-treated lymphocytes at 80-ms intervals, demonstrated a loss of optical density (P<0.001) suggesting a flattening of the cell but no significant increase in cell perimeter (P>0.05). Real-time multiparameter flow cytometry of melittin-treated lymphocytes confirmed simultaneous loss of the cytoplasmic marker, calcein, and uptake of the DNA dye, ethidium homodimer, but demonstrated no increase in forward light scatter. Transmission-electron microscopy of melittin-treated lymphocytes showed normal cell volumes but discontinuities in the cell membrane suggesting direct membrane toxicity. We conclude that melittin causes lymphocyte death by a "leaky patch" mechanism that is independent of colloid-osmotic effects.

Animals↗

Respiratory impedance following bronchoscopic or surgical lung volume reduction for emphysema.

BACKGROUND: Bronchoscopic methods for achieving lung volume reduction (BLVR) are presently undergoing clinical trials, and will soon be clinically available. Understanding the differential effects of surgical volume reduction therapy (LVRS) and BLVR on lung and chest wall physiology will assist physicians in selecting an optimal approach for patients. OBJECTIVES: Determine whether LVRS adversely affects lung or chest wall physiology at 3-month follow-up relative to BLVR in an experimental model of sheep emphysema. METHODS: Twelve mixed-breed sheep were treated with papain to produce experimental emphysema, and were divided into control, LVRS, and BLVR treatment groups. Lung and chest wall impedance was measured at 0, 5, and 10 cm H2O positive end-expiratory pressure at baseline and 3-month follow-up. RESULTS: Emphysema was associated with increased airway resistance, decreased lung tissue resistance and elastance, and increased chest wall tissue resistance. Following treatment, equivalent increases in lung elastance occurred in the LVRS and BLVR groups compared to controls. LVRS did not adversely affect chest wall impedance despite causing extensive pleural scarring. CONCLUSIONS: (1) Experimental emphysema following prolonged papain exposure progresses after cessation of treatment. (2) BLVR and LVRS produced equivalent lung and chest wall impedance responses at 3-month follow-up. (3) LVRS did not adversely affect chest wall impedance despite being associated with extensive pleural scarring.

Animals↗