Differentiated thyroid carcinomas associated with chronic thyroiditis: biological and clinical properties.

AIM: To determine biological and clinical features of papillary and follicular thyroid carcinomas associated or not associated with chronic thyroiditis. MATERIALS AND METHODS: The study was conducted by retrospective analysis of medical histories of 2,459 patients with thyroid cancer. Tumor size, its category according to the TNM system, multi-focal properties of tumor growth, carcinoma invasiveness, as well as disease stage, rates of relapses and metastasis, and also cumulative survival rates were analyzed. RESULTS: The tumor size in patients with papillary or follicular carcinoma associated with thyroiditis was smaller compared to the patients without thyroiditis. In the first case, the invasion frequency into extrathyroid structure and into the capsule was also lower. Multi-focal growth of both carcinoma types was registered more frequently in the presence of thyroiditis. The frequency of papillary carcinoma metastasis to lateral cervical lymph nodes was lower in the presence of thyroiditis, the frequency being equal for metastasis into lymph nodes of the VI lymph outflow zone in both groups of patients (with and without thyroiditis). In the presence of thyroiditis, the frequency of distant papillary carcinoma metastasis was decreased, no metastases were detected in patients with follicular carcinoma. In the group of patients with papillary carcinoma there was found no relation between the presence of thyroiditis and disease stage, relapse rates, and mortality levels; however, the risk of follicular carcinoma relapse was significantly lower in patients with thyroiditis. CONCLUSIONS: The presence of chronic thyroiditis in papillary carcinoma patients showed a certain positive impact on the course of the disease, in particular, primary tumor growth, invasion, and metastasis. Such effect is even more expressed in the patients with follicular thyroid carcinoma.

NITRIC OXIDE SYNTHASE ACTIVITY AND ITS CONCENTRATION IN THE TISSUES OF HUMAN THYROID CARCINOMAS.

The study of the activity of the constitutive form of nitric oxide synthase (cNOS) revealed that in the papillary thyroid carcinomas it corresponded to that detected in unchanged extratumoral tissue, while the enzyme activity in follicular carcinoma was half lesser. At the same time, the activity of inducible nitric oxide synthase (іNOS) was higher in the papillary and follicular carcinomas. Such changes in the enzyme activity were associated with an increase in its level in papillary carcinomas, and with minor changes in follicular carcinomas. In medullary carcinomas the parameters under study corresponded to those in unchanged tissue, and in the papillary carcinoma metastases without changes in enzyme activity of nitric oxide formation, the level of the latter was much higher. Elevated levels of nitric oxide and іNOS activity in papillary thyroid carcinomas did not depend significantly on the aggression characteristics of the latter, being however absent in tumors of T4 category on a background of reduced cNOS activity and less expressed in tumors surrounded by the tissue in the presence of a chronic thyroiditis. Furthermore, in the papillary carcinomas of papillary or follicular structure nitric oxide level did not differ from the normal range, being slightly higher in tumors of solid or heterogeneous structure with presence of solid areas, whereas in carcinomas of papillary-follicular structure it was twice, and in tissue of solidinsular structure three times higher. іNOS hyperactivity was observed in the carcinomas of different structure, except for tumors of solid structure, in the tumor of which enzyme activity was within the normal range, and in tumor of solid-insular structure where it was significantly higher (as well as cNOS activity) compared with tumors of other structure. Nitric oxide generating system is involved in the transformation of thyroid cells and progression of tumor growth, including through apoptosis regulation, as shown by the results of an analysis of data obtained both in the present study and previously. The nature of such involvement in papillary thyroid carcinomas with different histological structure is different. Key words: nitric oxide; constitutive and inducible nitric oxide synthase; thyroid carcinoma; apoptosis.

[Changes in active cysteine cathepsins in lysosomes from tissues thyroid papillary carcinomas with various biological characteristics].

To clarify possible role of cysteine cathepsin H, B and L in the proteolytic processes that contribute to the progression of tumor growth in the thyroid, we studied their activity in lysosomes isolated from the tissue of papillary carcinomas. It was shown that for these enzymes there is a dependence of the changes in their activity on a number of biological characteristics of the tumors. Thus, the sharp increase in the activity ofcathepsin H observed in lysosomes of tissue carcinomas category T2 and T3, with intra-and ekstrathyroid and lymphatic invasion of tumor cells. An increase in the activity of cathepsin B is set in the lysosomes of tissue heterogeneous follicular structure, especially in the presence of solid areas, in comparison with typical papillary tumors and in the lysosomes of tissue carcinomas in intrathyroid and cathepsin L-at extrathyroid invasion. A common feature of the enzymes is to increase the activity of cathepsins in lysosomes of tissue nonencapsulated papillary carcinomas. These enzymes probably do not take part in the invasion of tumor cells into blood vessels and in the mechanisms of tumor metastasis to regional lymph nodes. The latter shows no changes in the activity of cathepsins in lysosomes of tissue carcinomas category N1. The results indicate the different role of cathepsin H, B and L in thyroid carcinogenesis, where each enzyme has its specific function.

Abnormalities of apoptosis of the thyroid gland cells from extratumoral microfollicular tissue.

AIM: The aim of this study was to evaluate the frequency of existence of thyroid extratumoral normo- and microfollicular tissue in patients with thyroid carcinoma and peculiarities of apoptosis in mentioned tissue. MATERIALS AND METHODS: Using samples of normo- and microfollicular thyroid tissue it was determined the content of fragmented DNA and intensity of stimulated internucleosomal DNA fragmentation; activities of caspase-3 and cysteine lysosomal cathepsins. RESULTS: It was found that normofollicular tissue is observed more often in patients with nodal euthyroid goiter but microfollicular tissue is more common for patients with carcinoma. Extratumoral microfollicular tissue was found in the thyroid of patients above 50 years old mostly, and more rarely in young ones. The fragmented DNA concentration in microfollicular tissue was lower by a factor of 3.5 and intensity of stimulated internucleosomal DNA fragmentation was also decreased. Activity both of cathepsin B in lisosomes and caspase-3 in lysates of such tissue was also decreased. CONCLUSIONS: The decrease of intensity of spontaneous apoptosis and the absence of its modulation/induction following proapoptotic factors in extratumoral microfollicular thyroid tissue may be considered as a respond of the thyroid gland tissue to an existence of carcinoma.

[B-, H- and L-cathepsin-like activity in blood plasma of patients with diseases of the thyroid, parathyroid and adrenal glands].

B-, H- and L-catepsine-like activity regarding Na-benzoyl-D,L-arginine-4-nitroanilide, L-leucine-4-nitroanilide and azocasein was studied in the blood plasma of patients with different diseases of thyroid, parathyroid and adrenal glands. It has been shown that the high H- catepsine-like activity confirming the activation of blood catepsin H secretion accompanied by the tissue growth under any pathology took place in the blood plasma of patients with all the above diseases. High B- catepsine-like activity in the blood plasma of patients with thyroid diseases was exceptionally fixed under the nodular forms of thyroid pathology, while the character of changes in L catepsine-like activity under these diseases was not so natural. In diaseses of the adrenal glands the changes in the B- and L-catepsine-like activity were only shown in the blood plasma of patients with cerebral layer tumors but not the gland cortex: B-catepsine-like activity increased in the blood plasma of patients with benign or malignant tumors, and L-catepsine-like activity decreased under benign tumor from chromaffin tissue. The established picture of changes in enzymatic activity in the blood plasma of patients does not allow to think that the determination of blood B- and L- catepsine-like activity can be recommended for obtaining additional information at diagnosis thyroid diseases.

[Cathepsin-like activity in blood plasma of patients with papillary thyroid carcinoma].

The peculiarities of the thyroid changes depending on clinical characteristics of disease and histological structure of tumor were fixed by the studies of cathepsin-like activity in blood plasma of patients with papillary thyroid carcinoma. H-cathepsin-like activity had a tendence to more significant increase especially in patients with concomitant benign thyroid disease with increasing the tumor category. In a case of metastatic tumor without other concomitant thyroid pathology, the degree of an increase in enzymatic activity was significantly higher comparing to the blood of patients without tumor metastases. B-cathepsin-like activity in blood plasma of patients only with tumor carcinoma was at the level of that of healthy subjects. B-cathepsin-like activity in blood plasma had especially significant increased under tumor T1 category that was connected with nodular goiter or adenoma in a case concominant carcinoma by other thyroid pathology. In patients with carcinoma that was diagnosed against a background of concomitant pathology, the insignificant increase in L-cathepsin-like activity was not dependent on clinical features of disease, while in a case of carcinoma without concomitant pathology a degree of its increase was certainly dependent on a category of tumor T, its histological structure and the presence of oxyphilic cell metaplasia. We failed to get an evidence linking the changes in cathepsin-like activity in blood with the processes of intra- or extrathyroid invasion and invasion into the lymphatic or blood vessels. The results of the studies do not provide a rationale to consider that the cathepsin-like activity in blood of patients with papillary thyroid carcinoma can be the valuable additional biochemical criterion for differential diagnosis, estimation of the severity, and other characteristics of the disease.

[Mechanism of apoptosis in the thyroid cells in thyroid pathology].

The literature data and the results of own examinations concerning the changes in programmed cell death processes of goiter alterated thyroid tissue in patients with euthyroid or toxic goiter and also the extranodular tissue of the gland in the presence of pathological changes in it (the pronounced hyperplastic process, sclerotic and/or dystrophic changes, lymphoid infiltration, chronic or autoimmune thyroiditis) were analyzed. It has been shown that the significant changes in the processes ofmitochondria and postmitochondria regulation and apoptosis realization are occurred under thyroid pathology. The character and the degree of disorders in the apoptosis mitochondria mechanisms and the activity of destructive processes in the patient thyroid cells are dependent on many factors such as the type of pathology, the severity of its occurrence, the peculiarities of tissue follicular structure and the patient sex. Obtained resistance of the apoptosis mitochondria mechanisms can play an important role in the pathogenesis of thyroid nodular pathology or in the development of inflammatory and autoimmune processes.

[Effect of antioxidants on internucleosomal DNA fragmentation in thyroid tissue of patients with various thyroid pathology].

The in vitro effects of a-tocopherol and melatonin on the intensity of DNA internucleosomal fragmentation in tissue and neoplastic tissue of patients with thyroid pathology was studied. alpha-Tocopherol at a concentration of 10(-7) M and melatonin at a concentration of 10(-3) M were shown to inhibit internucleosomal DNA fragmentation in unchanged thyroid tissue, while in pathologically changed tissue and neoplastic tissue the effect of antioxidants was either absent, or those effects of preparations were prevailing, which were aimed at an increase in the intensity of internucleosomal DNA fragmentation. Some peculiarities of changes in the content of certain oligonucleosome fractions depending on the preparation, its dose, and thyroid disorder have been identified. The possible mechanisms of the effect of antioxidants on internucleosomal DNA fragmentation in normal thyroid tissue and in the presence of thyroid pathology are discussed.

[Glutamate decarboxylase activity and GABA binding in the brain synaptic membranes in rats with compensatory activation of hypothalamo-pituitary-adrenal system in stress]. / Hlutamatdekarboksylazna aktyvnist' i zv'iazuvannia HAMK synaptychnymy membranamy mozku shchuriv za diï stresu na tli kompensatornoï aktyvatsiï hipotalamo-hipofizarno-adrenokortykal'noï systemy.

The reaction of the rat adrenal cortex and medulla to stress was absent under conditions of compensatory activations of function of hypothalamic-pituitary-adrenal system. At the same time, glutamate decarboxylase activity was increased in the hypothalamus, decreased in the hyppocampus, and remained unchanged in medulla oblongata and cortex of cerebral hemispheres of hemiadrenalectomized rats after stress as compared to rats without stress. On the contrary, the intensity of specific GABA binding by synaptic membranes of medulla oblongata of hemiadrenalectomized rats was increased in response to stress effect it was decreased in the cortex of cerebral hemispheres, and remained unchanged in the hyppocampus.

[Gamma-aminobutyric acid in some non-neural tissues]. / Hamma-aminomasliana kyslota u deiakykh neneironal'nykh tkanynakh.

In this review modern data are presented on localization of GABA, functional characteristics and structure of the enzymes of GABA metabolism, transporters, and different types of GABA receptors in kidney, bladder, heart, blood vessels, spleen, skin and muscles.

[Glutamate decarboxylase activity and GABA binding by synaptic membranes of brain structure under stress in intact rats and rats with hormonal blockade of the hypothalamo-hypophyseal-adrenal cortex system]. / Glutamatdekarboksilazna aktyvnist' ta zv'iazuvannia GAMK synaptichnymy membranamy struktur mozku za umov stresu v intaktnykh shchuriv ta shchuriv iz gormonal'noiu blokadoiu sistemy gipotalamus-gipofiz-kora nadnyrkovykh zaloz.

Under the inhibition of hypothalamo-pituitary-adrenal system function, as a result of prolonged hydrocortisone administration, the normal neurochemical reaction to stress, i.e. stimulating the formation of GABA (mediator that is basic among responsible ones for the realization of stress-limiting brain function) was absent. In the medulla under such conditions an increase in the degree of GABA reception processes activation of comparing with the intact stressed rats was revealed and in the hippocamp of their changes was directivity reverse.

[Specific binding of GABA in the adrenal glands and catecholamine blood and adrenal gland levels in rats under stress with inhibited functioning of the hypothalamus-hypophyseal-adrenocortical systems]. / Spetsyfichne zv'iazuvannia GABA u nadnyrkovykh zalozakh i vmist katekholaminiv u krovi ta nadnyrkovykh zalozakh pry stresuvanni shchuriv za umov hal'muvannia hipotalamo-hipofizarno-adrenokortykal'noï systemy.

Forced swimming of intact rats was accompanied by an activation of the hormonal link in sympatho-adrenal system and by a decrease in the level of specific binding of 14C-GABA by adrenal plasma membranes; the former was prevented with the administration of GABAergic preparations (baclofen, GABA-taurine) before stress. Under inhibition of the function of hypothalamo-pituitary-adrenocortical axis following multiple hydrocortisone administration and, in presence of developing metabolic disorders and adrenal secretion of catecholamines, the latter reacted to stress by a decrease in adrenalin, noradrenaline and dopamine secretion without changing the intensity of specific binding of GABA. Administration of GABAergic preparations before stress prevented the development of stress disorders of noradrenaline and dopamine secretion partially or completely, and sodium thiosulphate administration increased acutely specific binding of GABA with adrenal plasma membranes simultaneously preventing the accumulation of adrenal noradrenaline and the decrease in blood DOPA level.

[Changes in the level of gamma-aminobutyric acid in the blood of children with thyroid diseases]. / Zminy vmistu hama-aminomaslianoï kysloty v krovi ditei z zakhvoriuvanniamy shchytovydnoï zalozy.

Blood GABA level was studied in children with thyroid disorders. Increase of GABA level was revealed in thyroid hyperplasia (11 degrees) and euthyroid goiters; in blood of children with diffuse toxic goiter there changes are much more significant. In children with thyroid cancer dramatic increase of GABA content was observed; in the nearest time following thyroidectomy blood GABA level decreased to low values, several mouths later it became normal, in a year and more it became elevated again. In blood from three children with congenital hypothyrosis the level of the mediator was decreased; no changes in GABA level were observed in children with autoimmune thyroiditis. Two hours after falling asleep blood GABA content lowered both in normal children and those with euthyroid goiter and thyroid cancer. The mediator level elevated sharply after finishing physical exercises in normal children and those with euthyroid goiter, while in children with thyroid cancer response of GABAergic system to physical exercises was opposite and in operated children it was absent.

[The GABA-ergic system parameters of the adrenal cortex in animals with normal and stimulated steroidogenesis]. / Deiaki parametry HAMK-erhichnoï systemy kory nadnyrkovykh zaloz tvaryn u normi ta za umov stymuliatsiï steroïdohenezu.

Activity of glutamate decarboxylase, a GABA synthesis enzyme, and intensity of its reception in the adrenal cortex and hypothalamus of guinea pigs and rats with normal and stimulated steroidogenesis was investigated. It has been shown that in the adrenal cortex there is a metabolic system which provides GABA synthesis from glutamate and mechanisms of GABA reception by plasmatic membranes. Mediator synthesis in the adrenal cortex is subjected to seasonal changes, GABA synthesis and reception selectively vary with administration of ACTH, prolactin and maintenance of animals on a diet with an excess of potassium ions.

[Change in the content of free amino acids in brain tissue and blood plasma of rabbits under the effect of hydrocortisone]. / Zmina vmistu vil'nykh aminokyslot u tkanyni mozku ta plazmi krovi kroliv pid vplyvom hidrokortyzonu

The influence of a single and long term intramuscular hydrocortisone injections on the content of free amino acids was studied in the rabbit brain tissue and blood plasma. It is shown that exogenous hydrocortisone changes the content of certain amino acids in the brain. The changes in the free amino acids spectrum in blood plasma are more significant than in the brain under the hydrocortisone effect. The direction of the changes in blood plasma at the single and long term hormone administration is the same, their degree of expression depends on the time of administration.