[Determination of amyloid P component in blood by an immunoenzyme method]. / Opredelenie amiloidnogo P-komponenta v plazme krovi s pomoshch'iu immunofermentnogo metoda.

An ELISA procedure was developed for measuring serum amyloid P-component (SAP). The assay is based on our finding of binding AP to polystyrene microtiter plates. The amount of SAP is determined by concurrent ELISA, wells being sequentially incubated with rabbit anti-AP antiserum and goat anti-rabbit antiserum, conjugated with peroxidase. The limit of sensitivity of the assay is 0.5 micrograms/ml. When applied to the screening of patients plasma, elevation of SAP concentration in patients with rheumatoid arthritis and amyloidosis were found.

[Partial purification of endogenous inhibitors of (+)-[3H] SKF 10047 binding with sigma-opioid receptors of the liver]. / Chastichnaia ochistka éndogennykh ingibitorov sviazyvaniia (+)-[3H] SKF 10047 s sigma-opioidnymi retseptorami pecheni.

Porcine liver extract inhibits (+)-[3H]SKF 10047 binding to rat liver membranes. The inhibitors of (+)-[3H]SKF 10047 binding were detected in acetone supernatant and in acid acetonated powder of the liver extract. Partial purification of the acetone supernatant revealed some characteristics of these inhibitors: low molecular weight (less than 1000), thermostability, resistance to pronase digestion, solubility in water and organic solvents.

[Limited proteolysis of prolactin in secretory granules from the pituitary gland of estrogenized rats]. / Ogranichennyi proteoliz prolaktina v sekretornykh granulakh iz gipofiza éstrogenizirovannykh krys.

The LTH-converting proteolytic activity in LTH granules isolated from estrogenized rat hypophysis was studied. Suspensions of granules were incubated at different values of pH for 4 hours at 37 degrees C. The reaction was controlled by SDS electrophoresis. Intensive proteolysis of LTH was observed at pH 6.0 and 3.9, which was accompanied by the formation of fragments with Mr 10, 12 and 17 kD and probably of smaller peptides. An inhibitory analysis revealed that the formation of the 17 kD fragment at pH 3.9 was partly and selectively inhibited by chloroquine, phenanthroline and phenylmethylsulfonyl fluoride. Pepstatin A fully inhibited the proteolysis, whereas leupeptin had no inhibiting influence. The data obtained testify to the presence in the granular fraction of the endopeptidase LTH-converting activity which is sensitive to pepstatin A, an aspartyl proteinase inhibitor as well as to chelators and a serine proteinase inhibitor.

[Study of the protein-peptide composition of secretory granules of bovine lactogenic hormone]. / Issledovanie belkovo-peptidnogo sostava sekretornykh granul laktogennogo gormona byka.

A combination of gel chromatography, fluorimetric analysis and polyacrylamide gel electrophoresis with immunochemical identification, the protein-peptide composition of secretory granules of the lactogenic hormone (LTH) isolated from the anterior lobe of bovine hypophysis was investigated. It was found that the peptide content of the granules is less than 3% of that of immunoreactive LTH. Using gel chromatography, the secretory granules were found to contain a hormone monomer and two immunoreactive forms with Mr 42 and 64 kD. With respect to immunoreactivity, the hormone form content was 90, 3 and 7%, respectively. Using polyacrylamide gel electrophoresis and subsequent immunochemical identification, the presence of four immunoreactive forms of LTH were identified in the secretory granules.

[Simple, highly specific procedures for radioimmunochemical estimation of hypophyseal protein hormones]. / Prostye vysokospetsifechnye metody radioimmunokhimicheskogo opredeleniia belkovykh gormonov gipofiza.

Immunosorbents, prepared by means of polymerization of antiserum proteins using chlorcarbonic acid isobutyl ester, were used to increased reproducibility and simplicity of the radioimmunochemical procedures for estimation of the estimation of the hypophyseal hormones. In the types of the radioimmunoassay (RIA) studied the immunosorbents were prepared by means of polymerization of the first and second antibodies. The universal immunosorbent, suitable to estimation of various antigens, was prepared from the second antibodies. The procedure enabled to use as the first antibodies the blood sera with comparatively low antibody titre. High specificity of the RIA procedures described was demonstrated in estimation of bovine hypophyseal hormones - lactogenous (LTH), somatotropic (STH) and beta-lipotropine. The calibration curves were linear in the concentration range from 2 to 20 ng/ml for LTH, and from 1 to 10 ng/ml for STH and beta-lipotropine. Estimation of LTH by means of the RIA procedure required 3 hrs, and of STH and beta-lipotropine - 16 hrs.

[Bovine epiphyseal lactotropin, biologically and immunochemically related to pituitary prolactin]. / Laktotropin iz épifiza byka, biologicheski i immunokhimicheski rodstvennyki prolaktinu gipofiza.

A substance biologically and immunologically related to the hypophysis lactogenic hormone (HLH) was detected in a purified preparation obtained by extracting the bovine pineal glands (epiphyses) with acidified acetonic saline fractionation, precipitation in isoelectric point and gel filtration. Bioactivity of the pineal gland lactogen was evaluated according to its ability for stimulating the pigeon thyroid gland proliferation. Equal doses of the pineal gland hormone and HLH produced the same biological effect. The lactogen immunochemical properties were studied by gel precipitation in agar gel, immunoelectrophoresis and radioimmunoconcurrent technique. Absolute identity of the hormone immunochemical properties was revealed. Molecular weights of both lactogens, determined by chromatography on Sefadex C-75, were equal. The average HLH content in the organ, determined by radioimmunochemistry, was 4.7 X 10(5) micrograms for the hypophysis and 9.6 X 10(2) micrograms for the pineal gland. HLH content, calculated per 1 mg protein, was 3 X 10(3) micrograms for the hypophysis and 24 micrograms for the pineal gland. The hormone content in the epiphysis was 1000 times higher than HLH concentration in the blood serum.

[[125I]-antigammaglobulin antibodies as universal reagents in radioimmunochemical methods of investigation]. / [125I]antigamma-globulinovye antitela kak universal'nyi reagent v radioimmunokhimicheskikh metodakh analiza.

A new indirect radioimmunochemical method based on the use of [125I]-anti-IgG-antibodies as universal detecting reagents is proposed. Its first step consists in the antibody binding to the antigen to be analyzed; the second step -- in immunoadsorption of the non-bound antibodies by water-insoluble sorbents prepared by chlorocarbonic acid isobutyl ester copolymerization of antigens with serum albumin or by immobilization of the antigen on Sepharose. The third step is the determination of the amount of sorbent-bound antibodies by means of [125I]-anti-IgG-antibodies. The method proposed was used for quantitative estimation of prolactin, somatotropin, lutropin, BB-isoenzyme of human creatine phosphokinase, testosterone and 5 alpha-dihydroxytestosterone. The method does not employ labelled antigens and is highly sensitive and highly specific.

[Gonadotropin from the bovine pineal body. Evidence of its similarity with the luteinizing hormone of the pituitary gland]. / Gonadotropin iz pineal'nykh zhelez byka. Dokazatel'stva identichnosti s luteiniziruiushchim gormonom gipofiza.

Gonadotropin affiliated by biological and immunochemical properties to the luteinizing hormone of the hypophysis was revealed in the purified extracts from the bovine pineal glands. Biological properties of gonadotropin from the pineal glands were assessed by the reactions specific of the luteinizing hormone. Immunochemical properties of gonadotropin were studied by means of gel-precipitation in agar gel, immunoelectrophoresis and radioimmunochemical analysis. It was found that the specific content of gonadotropin in the epiphysis was 80 times lower than the specific content of the luteinizing hormone in the hypophysis, but exceeded the LH concentration in the blood serum 555 times and in the cerebrospinal fluid 6.8 times. Immunoreactive LH was absent in the extracts of the brain cortex, the liver and the ovaries. A theoretical interpretation of a possible physiological role of the LH-like gonadotropin in the endocrine regulation of the reproductive function of the organism is given in this work.