(1) Human trypsin was purified from human pancreatic tissue and used as an immunogen in the production of monoclonal antibodies. It was also used as a standard preparation of human trypsin in the screening of monoclonal antibodies and in the development of an immunoradiometric assay (IRMA). (2) Eight monoclonal antibodies were raised to human trypsin, two of which were subsequently employed in the development of an IRMA to measure immunoreactive trypsin (IRT) in blood and blood spots. (3) An antiserum to human trypsin was raised in sheep and employed in a sandwich ELISA. (4) Monoclonal antibodies were tested by a number of different screening systems including ELISA, immunoblotting and immunoadsorption assays with 125I-labelled human trypsin. (5) Six selected monoclonal antibodies were tested for use (a) as the solid phase antibody and (b) as the radioactive tracer in an IRMA, and against each other for compatability in an IRMA. (6) One combination of antibodies was chosen and experiments performed to determine (a) lack of competition of binding to antigen between the two and (b) positive binding of the combination to serum IRT. (7) An IRMA to measure IRT in blood and blood spots was developed using two monoclonal antibodies, 56/C5/33 (IgG2a) and 125I-labelled 55/A4/31 (IgM). (8) The detection limit of the assay with purified cationic human trypsin as the analyte was determined to be 0.8 ng trypsin. (9) The IRMA was performed with reconstituted blood containing standards of human trypsin (either blood or dried blood spots on filter paper). (10) There is potential for this assay to be further developed and employed as a routine screening assay to detect elevated concentrations of IRT in dried blood spots from newborn infants with cystic fibrosis.

The S100 zinc-binding protein, calprotectin, is a noncovalently associated anionic complex which incorporates two immunologically distinct polypeptides in a molecule of Mr 36.5 kDa. Calprotectin's abundance in neutrophils, where it constitutes 60% of cytosolic protein, suggests an innate host defence function. In vitroit is microbiostatic. The trace metals, iron and zinc, are essential to the growth of most microorganisms. During the acute phase response to infection in man, the marked decrease in plasma iron is associated with a reduction in microbial growth. The similar, though smaller decrease in plasma zinc during this response to infection, may also act as a protective measure. As most microorganisms appear not to have zinc retrieval agents analogous to siderophores for iron retrieval, pathogenic microorganisms may be more susceptible to zinc deprivation than to iron deprivation. Thus, we hypothesized that calprotectin's in vitro candidastatic activity may be mediated by zinc chelation and that this phenomenon may also occur, in vivo, in plasma. In Sabouraud's culture medium, we demonstrated that calprotectin, at concentrations as low as 10ml, has potent, pH dependent candidastatic activity associated with zinc chelation. In plasma from children with cystic fibrosis (CF) & infected children, we demonstrated increased plasma calprotectin concentrations (median: 1832 & 4753 g/l respectively) compared to controls (median: 685 g/l). Plasma calprotectin concentration correlated positively with serum C- reactive protein in infected children, reflecting an association with the acute phase response. In CF children, there were positive correlations between plasma calprotectin and plasma copper and between plasma calprotectin and the Northern score, reflecting associations with the acute phase response and the extent of lung involvement respectively.

Short-chain fatty acids (SCFAs) are anaerobic bacterial metabolites. Cystic fibrosis (CF) lung disease is a condition caused by mutations in the cystic fibrosis transmembrane conductange regulator (CFTR) gene and is characterized by persistent lung inflammation and bacterial colonization. We measured the concentrations of SCFAs in sputum of patients with CF and tested the effect of these compounds on bacterial growth. Furthermore we found that SCFAs can influence the inflammatory protein expression and cytokine release in airway epithelial cells. SCFAs differentially alter cytokine release in CF bronchial epithelial cells (CFBE) compared to CFBE expressing wild-type CFTR. We also studied the effect of SCFAs in an acute lung injury model in BALB/cJ mice and found that intratracheally administered SCFAs can affect the inflammatory environment of the airways in vivo. We conclude that SCFAs may be important in the airways and that further investigation is warranted to understand their effects on inflammation and infection.

Short-chain fatty acids (SCFAs) are anaerobic bacterial metabolites. Cystic fibrosis (CF) lung disease is a condition caused by mutations in the cystic fibrosis transmembrane conductange regulator (CFTR) gene and is characterized by persistent lung inflammation and bacterial colonization. We measured the concentrations of SCFAs in sputum of patients with CF and tested the effect of these compounds on bacterial growth. Furthermore we found that SCFAs can influence the inflammatory protein expression and cytokine release in airway epithelial cells. SCFAs differentially alter cytokine release in CF bronchial epithelial cells (CFBE) compared to CFBE expressing wild-type CFTR. We also studied the effect of SCFAs in an acute lung injury model in BALB/cJ mice and found that intratracheally administered SCFAs can affect the inflammatory environment of the airways in vivo. We conclude that SCFAs may be important in the airways and that further investigation is warranted to understand their effects on inflammation and infection.

No description available.

No description available.

This study was designed to characterize the cardiac output (Q$ sb{ rm c}$) response to progressive submaximal upright (U) exercise in CF patients. Secondly, the Q$ sb{ rm c}$ adjustments were compared to those of similar supine (S) exercise, in an attempt to assess myocardial accommodation to the enhanced ventricular preload in the S posture. Q$ sb{ rm c}$ generally increased with exercise intensity in both U and S positions, although gr.IV plateaued at 50% VO$ sb2$max (S). Maximal stroke volume index (SI) was achieved at 50% VO$ sb2$max (U) in all groups, except gr.IV and at 30% VO$ sb2$max (S) in all groups. The change from U to S posture resulted in a significant (p $ le$.05) increase in SI at rest and for every submaximal exercise in gr.I, but not in CF patients, independent of disease severity eg(Rest:gr.I:27 $ pm$ 7(U) vs 39 $ pm$ 8(S); gr.II:24 $ pm$ 5vs28 $ pm$ 10; gr.III:18 $ pm$ 4 vs 22 $ pm$ 5; gr.IV:17 $ pm$ 4 vs 20 $ pm$ 6 ml/bt/m$ sp2$). These observations suggest a limitation in ventricular volume accommodation in CF patients which becomes apparent under the S exercise conditions.

Thesis (M.D.)--University of Adelaide, 1987. / Includes bibliographical references (leaves x-xxii).

Thesis (M.S.) in Microbiology--University of Maine, 2008. / Includes vita. Includes bibliographical references (leaves 52-58).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1967. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.