A radioimmunoassay method for the rapid detection of Candida antibodies in experimental systemic candidiasis

Rabbits were employed as experimental models to evaluate a solid-phase radioimmunoassay (RIA) method for the diagnosis of systemic candidiasis. Ten rabbits were inoculated subcutaneously to mimic superficial candidiasis and were found to produce no antibodies to Candida as determined by both immunodiffusion and RIA procedures. However, 94 per cent of 18 rabbits systemically infected by intravenous injection of Candida cells were observed to produce antibody as assessed by the RIA technique. These data encourage further tests with human sera and the continued development of this RIA procedure as a useful tool in the early serodiagnosis of systemic candidiasis.     

Interferon induction by mismatched analogues of polyinosinic acid . polycytidylic acid [(Ix,U)n.(C)n].

Interruption of the (I)n strand of (I)n.(C)n by unpaired bases [(U)] yielded mismatched analogues, (Ix,U)n.(C)n which were still effective as inducers of interferon, provided the I:U ratio (x) was equal or greater than 10. In highly sensitive interferon-induction systems such as primary rabbit kidney cells and human skin fibroblasts superinduced with cycloheximide and actinomycin D, (I10,U)n.(C)n and (I50,U)n.(C)n proved nearly as active as (I)n.(C)n. By virtue of their increased susceptibility to degradation by nucleases, (Ix,U)n.(C)n complexes with 10 less than or equal to x less than or equal to 50 may be expected not to persist as long in biological fluids as (I)n.(C)n, hence to induce fewer side effects.     

Na+,K+-ATPase: on the nature of the "cross-linked" subunits obtained in the presence of o-phenanthroline and cupric ion.

In previous studies on the quaternary structure of Na⁺,K⁺-ATPase, cupric-phenanthroline complex (CP) has been used for the cross-linking of the enzyme subunits. Here we show that the same products obtained in the presence of CP (α,α-dimer, α,β-dimer, and products of higher molecular weight) are also obtained when the enzyme is exposed to Cu²⁺ without o-phenanthroline. The α,β-dimer (but not the α,α-dimer) is dissociated in the presence of EDTA; indicating that this product is not the result of the covalent cross-linking of the subunits through a disulfide bond. The nature of the α,α-dimer remains to be determined. The findings suggest that the results of “cross-linking” experiments with CP should be interpreted with caution until the products are more clearly identified.     

Case report. Cervical carcinoma with selective IgA deficiency

A patient with cervical carcinoma was found to have selective IgA deficiency. The intact cell-mediated immunity, normal levels of IgG and IgM, and the absence of serum and salivary IgA established the diagnosis. Contrary to those of normal persons, salivary IgM was elevated and salivary IgA was not detectable in this patient. The patient had no signs attributable to IgA deficiency, but she always had dryness of the mouth. The association between cervical carcinoma and selective IgA deficiency was discussed.     

Two polypeptide chain constituents of the major protein of the cornified layer of newborn rat epidermis.

The insoluble component of stratum corneum of rat epidermis yields two major bands after extraction with 8 M urea-mercaptoethanol-dithiothreitol. The ratio of these two bands is about 1:1 in terms of protein stain intensity and S-[14C]carboxymethyl label. Both polypeptides were purified to homogeneity by DE-52-cellulose, sodium dodecyl sulfate hydroxylapatite C column chromatography, and preparative DodSO4-polyacrylamide gel electrophoresis. The heavier polypeptide contains 30% alpha helix and the lighter contains 27% alpha helix as determined by circular dichroism studies. Both are sensitive to Pronase and resistant to trypsin, collagenase, and elastase. The lighter chain is stable to pepsin but the heavier can be partially degraded to a smaller polypeptide with a molecular weight similar to that of light chain. Amino acid analysis shows that the light chain contains 12 more tyrosine residues than does the heavy chain, suggesting that the light chain is not generated from the heavy chain. However, the two chains may have a common peptide region. Antiserum prepared against the heavier polypeptide can be completely absorbed by purified lighter polypeptide and vice versa indicating that both chains have some common antigenic determinants. Antibody against either chain can cross-react with the stratum corneum and keratohyalin granules in the epidermis of newborn rat as indicated by fluorescent microscopic observation. Similarly, this antibody also cross-reacts with the cell surface or the contents of spinous and granular cells, and very weakly with basal cells, indicating that the two proteins may be present in the lower strata as well as the stratum corneum.     

(Na+ + K+)-activated adenosinetriphosphatase: fluorimetric determination of the associated K+ -dependent 3-O-methylfluorescein phosphatase and its use for the assay of enzyme samples with low activities.

Data are presented which prove that 3-O-methylfluorescein phosphate is a substrate for the K⁺-dependent phosphatase that is associated with Na⁺,K⁺-ATPase. Conditions for the continuous fluorimetric assay of 3-O-methylfluorescein phosphatase are described. Enzyme preparations from three different tissues with widely different specific activities exhibit similar K_m values for 3-O-methylfluorescein phosphate. Correlation between Na⁺,K⁺-ATPase activity and K⁺-dependent 3-O-methylfluorescein phosphatase activity is demonstrated in several partially purified enzyme preparations and crude tissue fractions. When the K⁺-dependent 3-O-methylfluorescein phosphatase of a crude rat-brain homogenate is assayed, the activity is a linear function of the amount of homogenate added to the assay mixture. The equivalent of 10 μg of brain tissue may be assayed under the conditions used. The potential value of this highly sensitive fluorimetric method for the assay of enzyme in small samples of various tissues is suggested.