Rapid microanalysis of cholesterol in bile and serum by gas chromatography.

Bile and serum cholesterol concentrations were analyzed by the same saponification, extraction and gas chromatographic procedure, utilizing stigmasterol as an internal standard and trifluoroacetate derivatization. The method was rapid and highly sensitive, requiring only 20 to 50 mul of sample. Comparable levels of precision were achieved by parallel analysis of a series of bile and serum samples by the present method and the Abell colorimetric method; a high degree of correlation was found between these two methods.

Characterization of a monoclonal antibody against concanavalin A binding antigen of Aspergillus fumigatus.

A monoclonal antibody was produced against a Concanavalin A (Con A) binding major epitope of Aspergillus fumigatus using a novel method of immunization. The antigen was purified using monoclonal antibody affinity chromatography reacted with specific antibodies present in human sera. Both allergic bronchopulmonary aspergillosis and aspergilloma showed high levels of antibody, against this purified antigen, when compared to normal controls. Similar results were obtained when the monoclonal antibody was used in a capture antigen assay. The antibody reacted with several A. fumigatus extracts in rocket electrophoresis demonstrating a single precipitin arc, which disappeared when Con A intermediate gel was used. This monoclonal antibody demonstrated reactivity only with cytoplasmic components of hyphae and spores of A. fumigatus, when a colloidal gold was used as a probe in immunoelectron microscopy.

Heart rate and oxygen consumption during walking on land and in deep water.

Oxygen consumption and heart rate were measured during treadmill walking and during walking in deep water in a group of women and in a group of men. In both groups predicted maximum oxygen uptakes were similar on land as in the water. However, there were obvious individual differences in the responses to land versus water exercise. These differences suggest that if deep water walking is used for training, the intensity of work should be assessed from preliminary fitness tests in water and not on land.

Monoclonal antibodies against farmer's lung antigens having specific binding to IgG antibodies.

Hypersensitivity pneumonitis resulting from environmental exposure to Saccharopolyspora rectivirgula (Micropolyspora faeni) among farmers has been well recognized. The diagnosis of the disease depends on demonstration of circulating antibodies against S. rectivirgula. However, dependable pure antigens are not available for serodiagnosis. In the present study we have employed hybridoma technology to obtain monoclonal antibodies against S. rectivirgula antigens. These monoclonal antibodies were employed to purify antigens through affinity chromatography. When tested in ELISA, high levels of antibodies were demonstrated against these antigens in farmer's lung patient sera compared to exposed but asymptomatic individuals from the same household. In Western blots patient sera reacted with components of crude antigens with molecular masses of 28, 35, 60, 65 and 68 kD and 4 components above 100 kD, while the monoclonal antibodies reacted only with the 60-kD protein. These purified antigens can be used as reliable reagents in the specific diagnosis of farmer's lung disease.

Cross-reactivity of food allergens in latex allergy.

Allergy resulting from exposure to latex proteins has been reported with increasing frequency in certain patient and occupational groups. Patients with latex allergy demonstrate cross-reactivity with some food allergens. Although amino acid homology of a few polypeptides from food and latex have been reported, no information is available comparing food and latex allergens. In the present study, we have obtained antibody from latex-sensitive patients by affinity absorption with various food and latex allergen extracts. The antibodies were then evaluated for reactivity with various antigens by ELISA. The results indicated that IgE cross-reactivity existed between different latex and food antigens. Hence, care should be exercised during evaluation of patients, as the clinical response may not be directed to the primary sensitizing antigen and may represent cross-reactivity of antigens.

Comparative studies of latex extracts used in skin testing.

BACKGROUND: There has been considerable variability reported in latex extracts used for skin testing and the results of those skin tests in the evaluation of latex allergy. Some test extracts have been associated with the induction of anaphylactic reactions in sensitive patients, while others have been reported as without adverse reactions. OBJECTIVE: The present study was designed to evaluate late extracts by comparing antigens from latex gloves. METHODS: Latex gloves were extracted for from 15 minutes to 24 hours for their protein contents and in vitro reactivities with IgE antibodies using sera of latex allergic patients and controls by ELISA. RESULTS: A higher yield of proteins was obtained after prolonged extraction, although reactivity of the sera by ELISA showed no major differences among the extracts. Extracts of 15 minutes elicited stronger reaction with health care workers and patients with spina bifida than with 24-hour extracts. The 15-minute extract demonstrated comparatively more reactive antigenic components in the fractions representing 3 to 10 and 10 to 30-kDa compared with fractions having less than 3 and over 30-kDa antigens. CONCLUSION: The study provided no evidence regarding the safety of one latex extract over the other related to time of extraction. The results indicated that a 15-minute extract can be used in ELISA for in vitro and, as in the literature, safe in vivo demonstration of latex specific IgE antibody.