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Biomedical subjects

W W Schultz

Publications and source records attributed to W W Schultz.

7 recordsLinked to original sources

Demonstration of specific antibodies to a coxsackie-like virus in patients of a hepatitis outbreak.

Lynchburg fecal virus (LFV), originally isolated from the stool of an infectious hepatitis patient, was passaged 15 times in WI-38 cells and partially characterized. Its properties are as follows: RNA virus; 27 nm in diameter, picornavirus-like morphology; inactivated at 56 C for 60 minutes; resistant to treatment with hydrochloric acid (pH 3.0), chloroform (33%), and diethyl ether (20%). Neutralization studies indicated that LFV is antigenically related to coxsackievirus A-24 but not to prototype hepatitis A virus (HAV). The simultaneous occurrence of infections with LFV and with HAV was indicated by the results of enzyme immunoassay (EIA) of the sera of patients of the Lynchburg outbreak. Absence of anti-LFV in patient sera was accompanied by an absence of anti-HAV and, conversely, an increased titer in anti-LFV was accompanied by an increased titer in anti-HAV. Each antibody type was shown to be elicited independently indicating simultaneous infections with LFV and HAV. The contribution, if any, of LFV to disease in the outbreak remains unknown.

Antibodies, Viral

Growth of Rickettsia typhi in irradiated L cells enhanced by lysosomal stabilization.

The growth of some obligate intracellular parasites is contingent upon avoidance of lysosomal activation during growth in their host cells. This is accomplished by the various parasites by different mechanisms and with different degrees of efficiency. The possibility was tested that the lysosomal stabilizer cortisone acetate might protect and thus enhance the growth of Rickettsia typhi in mouse L cells irradiated 6 days earlier. Beginning 2 days before infection of the L cells with a multiplicity of 10 rickettsiae, 20 microgram of cortisone per ml was added in medium 199 containing 5% fetal calf serum. This concentration of cortisone was below the cytotoxic level, as determined by viability staining, but was sufficient to significantly alter the ratios of cellular and released acid phosphatase and beta-glucuronidase in uninfected and infected cells, as shown by spectrophotometric analysis. Rickettsial replication, measured by hemolytic activity at 96 h and confirmed by microscopic observations at earlier stages of infection, was increased by the cortisone. Cortisone concentrations of 10 or 40 microgram/ml were less effective, and cortisone was ineffective when the rickettsial multiplicity per L cell was 2 or lower. These results indicate that amounts of cortisone that increase lysosomal stabilization in L cells favor rickettsial multiplication when the multiplicity of infection is relatively high.

Alkaline Phosphatase

Enzyme-linked immunosorbent assay for Pseudomonas aeruginosa exotoxin A.

An enzyme-linked immunosorbent assay (ELISA) is described for Pseudomonas aeruginosa exotoxin A. A double antibody sandwich method was used, employing polyvinyl microtiter plates as the solid phase, a primary coat of monospecific rabbit antitoxin serum, an outer layer composed of a horseradish peroxidase-sheep antitoxin immunoglobulin G conjugate, and an ortho-phenylene-diamine substrate. Absorbance (optical density) of hydrolyzed end product was read spectrophotometrically at 492 nm. ELISA detected as little as 30 pg (0.3 ng/ml) of purified toxin, and absorbance was linear over a 20-fold or greater concentration range. Toxin was demonstrated in culture filtrates from 42 of 48 (88%) consecutive clinical P. aeruginosa isolates compared with 37 of 48 (77%) positive by hemagglutination inhibition. Results of the two assays correlated closely (r = 0.82, P less than 0.001). Specificity was confirmed by neutralizability of ELISA activity with monospecific antitoxin. ELISA was thus a sensitive, specific, and quantifiable technique for the assay of P. aeruginosa exotoxin A in both purified and crude culture materials.

Enzyme-Linked Immunosorbent Assay

Virus-induced lysosomal enzyme dissolution of nasal turbinate cartilage.

The mechanism of laryngotracheitis virus-induced dissolution of chick nasal turbinate cartilage was studied by lysosomal enzyme histochemistry. Five-day-old chicks were infected by intranasal instillation, and changes in lysosomal enzyme distribution were followed at daily intervals through the tissue regeneration stage, Day 28. In the mucosa the lysosomes were activated beginning on Day 1, and glycerol acid phosphatase and a diffuse form of beta-glucuronidase were released concomitant with tissue cell destruction. In the chondrocytes (where glycerol acid phosphatase was absent), beginning on Day 2, particulate (lysosomal) beta-glucuronidase decreased as diffuse beta-glucuronidase increased and extended out into the matrix. The cartilage lost its metachromatic staining properties and became soft and pliable. Regeneration of the mucosa started on Day 6 and gradual reappearance of metachromatic staining of the cartilage began on Day 8 with considerable recovery of original turbinate structure by Day 12. A lysosomal membrane labilizer, vitamin A, exacerbated the cartilage pathology, whereas a stabilizer, cortisone, retarded it.

Acid Phosphatase

Effect of infection with gonococci on myeloperoxidase activity of leucocytes.

Lyosomal myeloperoxidase activity in human phagocytic leucocytes was stimulated by incubation with virulent (T1) and avirulent (T4) forms of Neisseria gonorrhoeae. The amount of activity, assayed by bacterial iodination (125-iodine) after 30 min. exposure to the pagocytes in the absence of serum, was about fifty times greater in cells infected with T4 strains. In the presence of heated human serum, or its IgG component, myeloperoxidase activity increased, but T1-Stimulated activity was significantly less than that of T4 and was not proportional to multiplicity of infection. From these results and from those of a previous study we conclude that T1 can stimulate leucocyte myeloperoxidase activity from an extracellular location, that for this activity a serum fraction is required, and that this may be a mechanism responsible for some of the killing of the membrane associated T1.

Humans

Hepatitis and operating-room personnel: an approach to diagnosis and management.

Hepatitis represents a common problem in operating room (OR) personnel. The differential diagnosis is usually narrowed to viral hepatitis versus halothane-associated hepatitis. While specific immunologic technics are available to diagnose viral hepatitis, halothane hepatitis cannot presently be unequivocally diagnosed with available clinical, biochemical, immunologic, or pathologic technics. Suggestions for management of OR personnel with hepatitis can only be based on insufficient evidence at present. The authors have initiated a prospective study to help clarify this situation.

Acute Disease