[The hypothalamo-hypophyseal system in patients with multinodular colloid euthyroid goiter]. / Sostoianie gipotalamo-gipofizarnoi sistemy u bol'nykh s mnogouzlovym kolloidnym éutireoidnym zobom.

Eighty patients with multinodular colloidal euthyroid goiter were examined. Thyrotropin-releasing hormone (TRH) test was carried out in 22 patients and 7 healthy women. The results of the test indicate a clear-cut tendency to reduction of hypophyseal TTH reserve in patients with multinodular euthyroid goiter with enlarged thyroid. In other words, clinical diagnosis of an euthyroid condition in the examinees appears to be groundless, particularly in patients with stage IV multinodular euthyroid goiter who may be referred to latent hyperthyrosis group on the basis of TRH test results. Three types of STH reaction were revealed by TRH test in these patients. The authors put forward a hypothesis on STH contribution as a growth factor to the pathogenesis of multinodular colloid euthyroid goiter.

[Humoral autoimmunity markers in autoimmune endocrine diseases]. / Markery gumoralnogo autoimmuniteta pri autoimmunnykh éndokrinnykh zabolevaniiakh.

To clarify the value of autoantibodies as risk factors of complications in various endocrine abnormalities, the incidence of autoantibodies to thyroid microsomal antigen (ATMA), thyroglobulin, and the surface antigens of the rat islet, adrenal cortex, adenohypophyseal cells and human skin fibroblasts was studied in patients with insulin-dependent mellitus (IDDM), at the onset of the disease and during one-year insulin therapy, non-insulin-dependent diabetes mellitus (NIDDM), Hashimoto thyroiditis, Graves' disease, diabetes associated with thyroidal dysfunction, euthyroid polynodular goiter, Schmidt and polyglandular syndromes and in the population. The antibodies were determined by ELISA. Polyclonal activation of the immune system was found in all abnormalities, except in polyglandular in children. The proportion of patients with more than one type of antibodies was minimal (26.4%) in IDDM and maximal (62.0%) in Graves' disease. Among IDDM patients, polyclonal activation of the immune system was observed more often in women than in men (48.5 vs 8.5%). The persistence of antibodies to fibroblasts in IDDM patients was associated with the development of vascular complications. The latter were observed in 4 of 7 patients who had these antibodies during a year and in none of negative patients. Thus, fibroblast antibodies may have a predicative significance for the development of late diabetic complications. The highest prevalence of these antibodies was discovered in Graves' disease (37.9%) wherein the antibodies may be involved in the development of exophthalmus and pretibial mixedema. Thyroidal dysfunction developed in all IDDM patients with ATMA preserved during a year and in none ATMA-negative patients.(ABSTRACT TRUNCATED AT 250 WORDS)

[Determination of autoantibodies to pancreatic beta-cells, non-islet endocrine cells and fibroblasts in patients with newly detected diabetes mellitus]. / Opredelenie autoantitel k beta-kletkam podzheludochnoi zhelezy, neostrovkovym éndokrinnym kletkam i fibroblastam u bol'nykh s vpervye vyiavlennym sakharnym diabetom.

The clinical onset of insulin-dependent diabetes (IDD) is characterized by the onset of circulation of autoantibodies to beta-cells. Thirty-three newly detected IDD patients and 14 newly detected patients with noninsulin-dependent diabetes mellitus were examined for autoantibodies to antigen P 64-69, to surface antigen of islet cells, to thyrocyte microsomal fraction, thyroglobulin, hypophysis, fibroblasts; the levels of circulating immune complexes were measured as well. IDD debut was found associated with the appearance of antibodies to pancreatic islet cells, thyroid, thyroglobulin, hypophysis, fibroblasts, this indicating a polyclonal activation of the immunity system. A relationship was revealed between antifibroblast antibody and anti-islet-cell antibody. Antihypophyseal antibodies were detected in 43% of patients with noninsulin dependent diabetes. Nine per cent of IDD patients and 24% of patients with noninsulin dependent condition were negative in the tests.

[An immunoenzyme method for determining autoantibodies to the microsomal antigens of human thyrocytes]. / Immunofermentnyi metod opredeleniia autoantitel k mikrosomal'nym antigenam tireotsitov cheloveka.

In order to investigate the role of autoantibodies in the pathogenesis of autoimmune thyroid diseases and for clinicoimmunological control of treatment, an enzyme-linked immunosorbent assay (ELISA) for the determination of autoantibodies against human thyroid microsomal antigens (TMA) was developed. ELISA was evaluated using sera from 156 patients with different thyroid pathology and diabetes mellitus and from 54 healthy donors. In parallel experiments the same sera were analyzed by an indirect immunofluorescence method for the presence of autoantibodies against TMA. Comparing all these data, both techniques appeared to have the same specificity, however ELISA was more sensitive than the indirect immunofluorescence method. Another advantage of ELISA is a possibility of long-term storage of microplates with immobilized antigen material without loss of antigenic activity. ELISA can be used for the determination of autoantibodies against TMA in human sera as a routine technique in clinical practice and experiments.

[Hypoinsulinemia, hyperglycemia and circulating antibodies to the islet cells during the development of streptozotocin diabetes in rats]. / Gipoinsulinemiia, giperglikemiia i tsirkuliruiushchie antitela k ostrovkovym kletkam pri razvitii streptozototsinovogo diabeta u krys.

The time course of metabolic parameters and islet cell surface antibodies (ICSA) in low-dose streptozotocin (STZ)-induced diabetes in rats was studied, a total STZ dose being 160 mg/kg body weight. Two-phase diabetes development was observed. Initial mild hypoinsulinemia and hyperglycemia turned to more severe diabetes after day 24 which was preceded by the first ICSA peak at day 13. The second ICSA peak occurred at day 35. The data obtained suggest that in this model of diabetes the toxic STZ effect induces both the diabetic syndrome and humoral autoimmunity to beta-cells, and the latter leads to further impairment of diabetes.

[The morphology of diabetic nephropathy in experimental diabetes induced by low doses of streptozotocin]. / Morfologiia diabeticheskoi nefropatii pri éksperimental'nom diabete, vyzvannom nizkimi dozami streptozototsina.

Kidneys of 16 Wistar rats were examined by light and electron microscopy, immunofluorescence and biochemically for the transamidinase activity at various periods of experimental diabetes induced by the fractionated intraperitoneal administration of low (40 mg/kg) doses of streptozotocin. 18 rats of the same age and sex served as control. This model of diabetes is characterized by a gradual decrease of the serum immunoreactive insulin, increase of hyperglycemia, the presence of "insulitis" 19 days after the beginning of the experiment and the development of nephropathy in the genesis of which immune mechanisms might participate. Transamidinase activity correlated with the alterations of renal tubuli. The conclusion is made on the possibility of using this model of experimental diabetes for studying the pathogenetic mechanisms of renal lesions in diabetes; transamidinase activity allows one to evaluate the nephron function in diabetic nephropathy.

[Molecular biological aspects of the pathogenesis of diabetes mellitus]. / Molekuliarno-biologicheskie aspekty patogeneza sakharnogo diabeta.

The discovery of the key role played by the immune system in the pathogenesis of insulin-dependent diabetes mellitus (IDDM) opens up new possibilities for its early diagnosis, at the stages preceding its clinical manifestation. Analysed are the markers of genetic susceptibility to IDDM associated with some major histocompatibility complex antigens (specifically with HLA-DR 3, HLA-DR4, and HLA-DQ), of the cellular and humoral anti-islet autoimmunity, as well as the origin of the islet-cell autoantigens. The markers' significance for the diagnosis and prognosis is discussed.

[Comparative study of physico-chemical and biological properties of human somatotropin produced by genetic engineering and isolated from the pituitary gland]. / Sravnitel'noe issledovanie fiziko-khimicheskikh i biologicheskikh svoistv somatotropina cheloveka, poluchennogo gennoinzhenernym metodom i vydelennogo iz gipofizov.

Human somatotropin hormono (STH), produced by means of gene engineering in the complex program "Human growth hormone", managed by the Academy of Sciences of the USSR, Ministry of Medical and Biological Industry of the USSR and Ministry of Public Health of the USSR, was shown to be similar in its physico-chemical properties to the main isoform of highly purified STH, isolated from human hypophysis. As distinct from the hypophyseal STH (STHhyp) containing minor isoforms of the hormone, the preparation of biosynthetic STH (des-Phe1-STH; STHbio) proved to be homogeneous. Studies of biological properties showed that STHbio exhibited high, similar to STHhyp, immunological, growth-stimulating and insulin-like activities as well as it possessed the lipotropic effect in vivo. The lipotropic effect of STHbio in vivo was less prolonged as compared with that of STHhyp, while in vitro it was only slightly expressed in isolated rabbit fat tissue. The effect did not depend on the hormone dose, apparently due to either absence of the hormone modified forms in the STHbio preparation or other hypophyseal contaminating substances responsible for the lipotropic activity. STHbio, similarly to STHhyp, did not stimulate DNA synthesis in blood serum-free culture of human fibroblasts. Studies of STHbio biological properties suggest that multifunctionality of native STHhyp appear to depend on intrinsic specificity of its molecule.

[The role of proteolytic processes in the stimulation of lipolysis in the adipose tissue by somatotropin, adrenocorticotropin and beta-lipotropin]. / Rol' proteoliticheskikh protsessov v stimuliatsii lipoliza v zhirovoi tkani somatotropinom, adrenokortikotropinom i beta-lipotropinom.

The influence of proteinase inhibitors on the lipotropic effect of somatotropic (STH), adrenocorticotropic (ACTH) and beta-lipotropic (LPH) hormones in adipose tissue was studied in vitro. The effect of STH was found to be completely dependent on the activity of tissue serine proteinases of trypsin and chymotrypsin types. The effect of LPH partly depended on serine proteinases of chymotrypsin type, whereas that of ACTH--on chymotrypsin and carboxylic proteinases. The effects of all the three hormones were also manifested during lysosomal proteolysis. The protease-dependent inhibition was specific for polypeptide hormones and was unobserved in the lipotropic effect of adrenaline. The inhibiting effect of serine proteinase inhibitors on hormones pretreated with blood plasma or proteinases was much weaker than on untreated hormones. In adipose tissue the early insulin-like effect of STH, unlike the late lipotropic effect, was independent of proteolysis. It was assumed that primary proteolysis plays a role in the activation of polypeptide hormones which is necessary for the manifestation of the lipotropic action.

[Effect of the synthetic tetradecapeptide corresponding to the amino acid sequence of human somatotropin 31-44 on rat blood and liver lipids]. / Vliianie sinteticheskogo tetradekapeptida, sootvetstvuiushchego aminokislotnoi posledovatel'nosti 31-44 somatotropina cheloveka, na lipidy krovi i pecheni krys.

Although synthetic tetradecapeptide, involving the 31-44 amino acid sequence of human growth hormone, exhibited a distinct lipolytic effect, content of triglycerides was not increased in rat blood and liver tissue during chronic experiments. At the same time, the level of atherogenic lipoproteins as well as activities of postheparin lipase and liver triglyceride lipase were also unaltered. The data obtained suggest that the tetradecapeptide studied stimulated simultaneously both lipolysis and oxidation of free fatty acids.

[Suppression of lipotropic action of human somatotropin by protease inhibitors]. / Podavlenie lipotropnogo deistviia somatotropina cheloveka ingibitorami proteinaz.

The effect of protease inhibitors on the lipotrophic action of the human growth hormone was studied in rabbits in vivo and in vitro. The human growth hormone at a concentration of 25-100 micrograms/ml stimulated lipolysis in isolated rabbit perirenal fat tissue 3-5 fold. An addition of 250 or 500 E Trasylol or 5 mM methylamine to the incubation mixture inhibited the growth hormone lipotrophic effect by 50%. Contrary to these protease inhibitors, the aminopeptidase inhibitor bacitracin did not affect the growth hormone action. Trasylol and methylamine did not diminish either basal or epinephrine-stimulated lipolysis in rabbit fat tissue. In vivo Contrical at the total dose of 5000 E infused 30 min before and 30 min after the growth hormone injection completely abolished the increase of plasma-free fatty acid level induced by the hormone. These data suggest that: 1) the inhibitor suppression of growth hormone lipotrophic action is not due to the inhibitor influence on lipolysis or on hormone action at the cell membrane level but is the result of direct protease inhibiton; 2) the late lipotrophic action of the growth hormone may require preliminary cleavage of the hormone molecule in which serine proteases may be involved.

[Chronic action of the fat-mobilizing fragment of human somatotropin on the fat content of fat depots and the plasma glucose level in rabbits]. / Khronicheskoe deistvie zhiromobilizuiushchego fragmenta somatotropina cheloveka na soderzhanie zhira v zhirovykh depo i vroven' gliukozy v plazme krovi u krolikov.

The effect was studied of synthetic tetradecapeptide-the fat-mobilizing human somatotropin fragment 31-34--on the fat content in fat depot and glucode concentration in the rabbit blood plasma and urine. A marked decrease in subcutaneous (the nape region), perirenal, mesenteric and epididimal fat content, accompanied by partial replacement of the fatty tissue by the connective one, was seen 3 to 4 weeks after tetradecapeptide injection to rabbits. Peptide effect manifested itself only under condition of the inadequate diet and was especially pronounced in young animals. Clucosuria and hyperglycemia were not observed after chronic tetradecapeptide injection. Tetradecapeptide did not exert an evident hyperglycemic or insulinogenous effect and did not produce a marked glucose tolerance fall after short administration to rabbits and during the glucose tolerance test in rats.

[Lipolytic effects of a synthetic tetradecapeptide, corresponding to 31-44 amino acid sequence of human somatotropin]. / Kharakteristika zhiromobilizuiushchego deistviia sinteticheskogo tetradekapeptida, sootvetstvuiushchego aminokislotnoi posledovatel'nosti 31-44 somatotropina cheloveka.

A synthetic tetradecapeptide 31--44 developed a fast lipolytic effect in rats and rabbits in vivo and in vitro. The concentration of plasma fatty acid was maximal after 30 min following the injection and came down to the initial level after 60--120 min. The effect was prolonged when the tetradecapeptide was injected in the form of Zn-suspension. Both in rats and rabbits the in vivo lipolytic effect of native somatotropin was pronounced only after a 2-hr latency. Actinomycin D and cycloheximide did not prevent the lipolytic effect of the tetradecapeptide in isolated adipose tissue; however, this effect was not exerted in the absence of Ca2+. The hexapeptide fragment of tetradecapeptide was inactive in rats in vivo, but had a slight effect an isolated rat adipose tissue when taken at high doses. Thus, the lipolytic effect of the tetradecapeptide requires Ca2+ and is more pronounced in rabbits than in rats. Contrary to somatotropin, the effect of tetradecapeptide is fast and independent of RNA and protein synthesis. These data and previously shown tetradecapeptide lipolytic effect on isolated human adipose tissue suggest that the tetradecapeptide possesses structural features required for the lipolytic effect of the whole hormone.

[Primary structure of cyanogen bromide fragments of sei whale (Balaenoptera borealis) pituitary somatotropin]. / Strukturaia kharakteristika bromtsianovykh fragmentov gipofizarnogo comatotropina kita vida seival (Balenoptera borealis)

5 fragments are isolated after the degradation of somatotropin from sei whale pituitary glands with cyanogen bromide: N-terminal 4-segmented; C-terminal 12-segmented with the internal disulfide bond; middle 25- and 30-segmented and a high molecular weight fragment following N-terminal tetrapeptide and bound with disulfide bond to 30-segmented fragment. Complete amino acid sequence of three shortest cyanogen bromide fragments is deciphered and N- and C-terminal sequence is investigated in two large fragments after their uncoupling under performic acid oxidation. Amino acid sequence is deciphered of a peptide obtained after trypsine hydrolysis of 30-segmented cyanogen bromide fragment. Comparison of amino acid sequence of whale somatotropin fragments with that of sheep, beef and human somatotropin has revealed that 57 out of 61 identified amino acid residues of whale somatotropin repeat amino acid residues in similar regions of beef somatotropin, 56--of sheep and only 42--of human somatotropins. Besdies, 4 of 5 revealed amino acid substitutions in whale hormone, as compared with sheep somatotropin, are amino acids which are present at the same positions in human hormone.

[Effect of synthetic tetradecapeptide corresponding to 31-44 amino acid sequence of growth hormone on lipolysis in human adipose tissue]. / Vliianie sinteticheskogo tetradekapeptida, sootvetstvuiushchego aminokislotnoi posledovatel'nosti 31--44 gormona rosta cheloveka, na lipoliz v zhirovoi tkani cheloveka

Fat mobilyzing activity of synthetic tetradecapeptide, which corresponds in 31--44 amino acid sequence of human growth hormone, is studied in vitro in human fat tissue. The peptide at concentrations of 3--33 microng/ml considerably stimulated lipolysis in subdermal fat tissue, omentum and shoulder ateroma. Minimal efficient peptide concentration was 3 microng/ml in most experiments, sometimes it was 0.3 microng/ml. Direct dependency between dose logarithm and lipolysis rate was observed at dose interval of 0.3--18 microng/ml. Native growth hormone produced no activity in human fat tissue even in concentrations of 50--100 microng/ml.

[Lipid-mobilizing activity of a synthetic peptide identical to 33-44 fragment of human growth hormone]. / Zhiromobilizuiushchaia aktivnost' sinteticheskogo peptida, identichnogo fragmentu 31-44 gormona rosta cheloveka

The lipid-mobilizing activity of a synthetic peptide, NH2-Phe-Glu-Glu-Ala-Tyr-Ile-Pro-Lys-Glu-Gln-Lys-Tyr-Ser-Phe-COOH, corresponding to the 31-44 amino-acid sequence of human growth hormone, was studied. The peptide stimulated lypolysis upon administration to fasted rats and during incubation with isolated epidiymal adiposed tissue of rat and perirenal adiposed tissue of rabbit. The lipid-mobilizing effect of the peptide,unlike the corresponding effect of the native growth hormone, developed fast and was markedly pronounced 15-30 min after the incubation was started. Direct dependence between the peptide dose logarithm and the effect studied was observed at concentrations of 0.01-10microng/ml during incubation with rat adipose tissue and at 0.001-0.1 microng/ml during incubation with rabbit tissue.