The isolation of the gamma subunit of fetal hemoglocin (HbF) and its use in a radioimmunoassay for HbF.

A method is described for the purification, from fetal hemoglobin (HbF), of the fetal specific globin chain (gamma chain) in its native state. In the absence of alpha chain (the globin chain common to all adult human hemoglobins) gamma chain, when used as an immunogen, is able to express its unique antigenicity. Here, a specific, high titer antiserum raised against gamma chain has been used to establish a sensitive radioimmunoassay for HbF. This approach may be applicable to the measurement of other normal and abnormal hemoglobins.

Double-blind trial comparing flunisolide and placebo for the treatment of perennial rhinitis.

The effectiveness of a new topical steroid, flunisolide, was evaluated for the treatment of patients with perennial rhinitis. The sixty-nine patients in this 8 week double-blind parallel trial received either 300 microgram/day of flunisolide or placebo. The flunisolide group showed statistically greater improvement than the placebo group in such symptoms as the duration of sneezing, stuffy nose, runny nose and nose blowing. 63% of the flunisolide group compared with 39% of the placebo group felt the test spray had provided substantial or total control of their nasal symptoms (P = 0.0026). When those patients who were skin test-positive (allergic) were considered separately from those who were skin test-negative (non-allergic), differences in responsiveness were noted. While those patients who had negative skin tests received greater relief of their symptoms with flunisolide than with placebo, patients who had positive skin tests showed a greater responsiveness to flunisolide and reported a higher incidence of significant or total control of their symptoms. Careful monitoring of plasma cortisol levels revealed no evidence of adrenal suppression in any of the patients in this trial. This lack of suppression may be due to two factors: (a) flunisolide is rapidly metabolized into a metabolite with a low degree of activity and (b) a very low dose of corticosteroid is administered.

Physicochemical approach to the purification of human alpha1-fetoprotein from the ascites fluid of a hepatoma-bearing patient.

A method for the purification of human alpha1-fetoprotein from the ascites fluid of a hepatoma-bearing patient is described that is capable of yielding large quantities of pure alpha1-fetoprotein within a relatively short period of time. The technique is based entirely on the physicochemical properties of the alpha1-fetoprotein molecule and uses sequential purification steps: ion-exchange chromatography on DEAE-Sephadex A-50, molecular-sieve chromatography on Sephadex G-200, negative-affinity chromatography on Sepharose-Blue Dextran, positivepaffinity chromatography on concanavalin A-Sepharose and, finally, molecular-sieve chromatography on Sephadex G-100. The efficiency of the entire procedure in its present form is 15% of the alpha1-fetoprotein activity of the starting preparation from ascites fluid. The purity of the final product was shown by polyacrylamide gel electrophoresis, radioimmunoelectrophoresis, and determinations of the NH2-terminal and COOH-terminal amino acid residues of the alphs1-fetoprotein isolated. Amino acid analysis of the final product revealed a composition very similar to those reported for alpha-fetoprotein preparations that have been previously isolated by the use of immunochemical technology.

Oncofetal antigens. Increasing the specificity of the CEA radioimmunoassay.

The biologic and clinical significance of the oncofetal antigens carcinoembryonic antigen (CEA) and alpha1-fetoprotein (AFP) are discussed. Although the current assays for these molecules are not tumor-specific, measurement of these molecules in the circulation of cancer patients is useful either for tumor diagnosis or for management of the cancer patient in the postoperative or post-chemotherapy state. An approach to increasing the specificity of the CEA radioimmunoassay is described.

Vascular involvement in relapsing polychondritis.

Review of four cases of relapsing polychondritis (RP) seen at one hospital in the 12-year period 1963 to 1974 revealed that one patient had aortic insufficiency with large artery involvement, two others had involvement of medium and large arteries and the fourth may have had mucocutaneous vasculitis. Valvular disease has occurred in 9% of all cases of RP reported in the literature and, if vasculitis beyong the aortic root is included, 25% of cases of RP manifested inflammatory vascular disease. The frequency of pseudotumour of the orbit and cochlear-labyrinthine dysfunction is also high and may be a manifestation of vasculitis.

The isolation and characterization of tumor-specific antigens of rodent and human tumors.

Putative tumor-specific transplantation antigens (TSTA) from both a carcinogen-induced rodent tumor (MC-1) and 2 human tumors were purified. The antigens were solubilized from the tumor cell membranes by limited papain digestion in a manner similar to that described for the isolation of normal histocompatibility antigens. The antitumor immune response of the tumor-bearing host was used to monitor the purification of the putative TSTA in both the rodent and human tumor systems. In the case of the rodent tumor, a major step in the purification of the TSTA involved affinity chromatography on Sepharose beads coupled to autologous antitumor antiserum. A comparable procedure was utilized in the purification of the TSTA from human tumors by using affinity chromatography on anti-human beta2-microglobulin antiserum coupled to a solid phase. The data obtained indicate that the TSTA of human tumors contains a beta2-microglobulin chain that is immunochemically identical with, and very similar in size to, that found in normal human histocompatibility antigens. A subunit of similar size was also identified in the carcinogen-induced rodent tumor. These results suggest that the TSTA in both humans and rodents may well be altered histocompatibility antigens.

New perspectives in allergic asthma.

Major advances have recently been made in both the diagnosis and the treatment of allergic asthma. The radioallergosorbent test (RAST), which measures allergen specificity of lgE antibodies in vitro, offers as good results as direct skin tests for allergy, without the inconvenience, discomfort and risk for the patient of the latter. However, the RAST must be done in a radioisotope laboratory, and standardized extracts of test allergens are lacking. Several new drugs, administered by inhalation and relatively free from serious side effects, are highly effective in treating asthma. Disodium cromoglycate, believed to stabilize tissue mast cells, protects particularly against exercise- or antigen-induced bronchospasm, and often permits reduction or withdrawal of corticosteroid therapy. Some acetonides or esters of glucocorticoids, because of their greatly increased topical anti-inflammatory activity, are effective in doses too small to cause serious systemic side effects. With the most topically effective, beclomethasone dipropionate, systemic corticosteroid therapy can usually be reduced or withdrawn, but recovery of hypothalamic-pituitary-adrenal function may be delayed; the drug regimens should therefore overlap.

Studies of the linkage relationship of beta-2-microglobulin in man-mouse somatic cell hybrids.

Beta-2-microglobulin (beta2mu) production has been studied in 33 primary man-mouse hybrid clones and in 26 secondary man-mouse hybrid clones. These clones have also been examined for the presence of 15 human enzyme phenotypes. Karyotypic analyses have been carried out on clones. From these studies the following conclusions can be drawn: (1) Gene(s) determining human beta2mu production in humans are apparently syntenic with the MPI gene on chromosome 15. (2) Long-term fibroblast lines may be of limited use in mapping studies as chromosomal rearrangements frequently occur in these lines. (3) The gene(s) determining beta2mu production in humans segregate independently of chromosome 6. If the assignment of the genes determining HL-A alloantigens to chromosome 6 is correct, our results imply that beta2mu and HL-A alloantigens are determined by genes carried on different chromosomes, despite the fact that beta2mu forms an integral part of the HL-A molecule.