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.

[Characteristization of the human microencapsulated adrenal cortex tissue in long-term culture].

Microencapsulated human adrenal cortex tissue preserves the ability to secrete 11-oxycorticosteroids for 51 days of cultivation, and to react adequately in responses to stimulation with adrenocorticotrophin and inhibition with chloditane, suggesting good prospects of the use of this tissue for compensation of the hypofunctional state of adrenocorticotropin system in experimental animals.

[Effect of N-stearoylethanolamine on the DNA fragmentation intensity in tumour and extratumoral tissues of the human adrenal cortex].

The effect of different concentrations of N-stearoylethanolamine (NSE 18:0) on fragmentation of DNA in the tumoural and extratumour tissues of the adrenal glands in vitro was studied. In this work the following types of tissue were investigated: extratumoural tissue from patients with hormonally active tumours, benign tumour tissue (hormonally active and hormonally inactive), tissue of malignant tumours and hyperplasic tissue of the adrenal glands (Itsenko-Cushing disease). It has been established that the NSE increases the intensity of DNA fragmentation only in the tissue of hormonally inactive tumours. Benign hormonally active tumours, malignant tumours and hyperplastic tissue of the adrenal glands were resistant to the NSE. The possible mechanisms of resistance to the drug are discussed.

[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.

[Analysis of the expression of RET/PTC oncogenes in post-chernobyl papillary thyroid carcinomas of patients from different age groups].

A comparative analysis of the expression of both, RET/PTC1 and RET/PTC3 oncogenes in papillary thyroid carcinomas (PTC) of patients from different age groups was carried out. Those were the following groups: children (mean age - 13 years, mean latency period - 13 years), young adults (mean age - 24 years, mean latency period - 14 years), adults (mean age - 38 years, mean latency period - 22 years). The presence of RET/PTC oncogenes was detected using polymerase chain reaction. In all cases the samples of both tumor and normal thyroid tissue were studied. It was established that induction of both, RET/PTC1 and RET/PTC3 rearrangements was present only in carcinoma samples. In PTCs the percentage of RET/PTC-positive tumors with increasing the age of patients has been decreasing. It should be noted that the part of carcinomas with induction of RET/PTC1 did not change with increasing the age of patients. At the same time the frequency of RET/PTC3 rearrangements with the increasing both the latency period and age of patients, significantly decreased. In conclusion, our data can evidence for the presence of correlation between the age of patients, latency period and induction of RET/PTC3 oncogenes.

[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.

[Effect of 17beta-estradiol on mRNA expression of Bax proapoptotic protein and DNA fragmentation level in the human adrenal cortex].

The effect of 17beta-estradiol on the change of proapoptotic protein Bax mRNA level and DNA laddering in human adrenal cortex tissue was studied. The adding of 17beta-estradiol to incubation medium caused a decrease of the proapoptotic protein Bax mRNA level in adrenocorticocytes by 22% in comparison with control. Hormone influence decreases the DNA fragmentation by 50%. The obtained data suggest that 17beta-estradiol caused a significant antiapoptotic effect in human adrenal cortex.

[Histological and hormonal characteristic of microencapsulated human parathyroid tissue in long-term culture].

Microencapsulated human parathyroid tissue preserves the viability and high ability to secrete parathormone in long-term culture that indicates on perspective to further study the efficiency of using this tissue in the presence of compensated hypofunctional state of parathyroid system in experimental animals.

[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.

[The influence of protein kinases ERK, JNK and nuclear transcriptional factor c-JUN on corticotropin signal transduction in adrenocortical cells].

The estradiol effect on the expression level of ERK1/2, JNK, p38 mitogen-activated protein kinases, c-Jun and c-Fos transcription factors in the adrenal cortex of male rats was studied. It was shown that a single corticotropin injection caused significant changes of the ERK1/2 protein kinases levels in the adrenal cortex. This index was 1.6 times higher 1 h after the injection and was decreased 6 h after the injection. The corticotropin influence on JNK level is very significant. The protein kinase JNK level was 2.4-fold increased I h after and was decreased 6 h after the injection. However, the level of p38 kinase was not changed in these conditions. The level of c-Jun transcriptional factor increased 1.7 times 1 h after corticotropin treatment and continued decreasing with the increase of time after the injection. The level of c-Fos factor increased 1.7 times only 6 h after the injection. These changes may be an important factor of activation of adrenocortical function provoked by corticotropin.

[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.

[Estimation of frequency of the main clinical features in patients with malignant tumor of thyroid gland after radical operative intervention].

The rate of main clinical features was studied in 25 patients with the thyroid gland carcinoma after performance of total thyroidectomy, it was compared also with such a rate in patients with stable hypoparathyroidism as a complication of radical operative intervention made on thyroid gland or parathyroid glands. There was established, that nearly all physical features and majority of somatic symptoms are caused by the stable postoperative hypoparathyroidism occurrence.

[Gender-related risk of non-fatal stroke, myocardial infarction and blindness in the type 2 diabetic patients depend on the type of treatment].

The aim of present observational cross-sectional study was the estimation of the gender-related risk of non-fatal stroke, myocardial infarction and blindness in type 2 diabetics (T2D) depending on treatment in Ukraine. In Dec 2005 the data from 11 Ukrainian regions for all living T2D already included into the nationwide population-based diabetes register (start of creation Jan 2001) was extracted. Male/female relative risks (RRs) for non-fatal stroke, myocardial infarction and blindness were calculated in treatment groups: Diet only (DO) 7273/15901; oral antidiabetic drugs (OAD) 15109/ 33913; insulin (I) 5529/12462 male/female respectively. 95% confidence intervals were estimated using logarithmic transformation, chi-square test was used to compare the differences between groups. The higher prevalence of stroke among men was in those patients treated with OAD or DO:RRs = 1.38 (1.26-1.51) and 1.48 (1.25-1.76), respectively. The gender differences were absent among those treated with I:RR = 0.98 (0.87-1.10). An excess male risk of myocardial infarction was in persons treated with I:RR = 1.37 (1.21-1.55), though smaller than in those treated without I - male/female RRs: 2.18 (1.97-2.41) and 2.37 (1.99-2.86), respectively for OAD and DO (P<0.001 in both cases). The significantly increased female prevalence of blindness was in patients treated with I:RR = 0.80 (0.70-0.92). The latest is in accordance with our earlier results, which have demonstrated the raised prevalence of both low sight and proliferative retinopathy among the insulin-treated diabetic women. No sex-related differences in blindness prevalence were revealed in patients treated with OAD or DO. The gender risks of not-fatal stroke, myocardial infarction and blindness in T2D are significantly different depending on the fact of insulin treatment. Further research could clarify if it depends upon insulin-related hyperandrogenisation in T2D women.

[Effects of estradiol on synthesis of corticosteroids, protein kinase A and C activity in the adrenal cortex of intact and orchiectomized rats].

Estradiol treatment produced significant increase of total 11-hydroxycorticosteroids level in the blood of intact and castrated rats. Activity of protein kinase A increased in the cytosol and membrane fraction of adrenocorticocytes of intact and orchiectomized rats after estradiol influence. Activity of protein kinase C significantly raised in the cytosol and membrane fraction of adrenocortical cells in all investigated groups. Our results suggest that cAMP-dependent protein kinase A and protein kinase C mediate estradiol effects in adrenal cortex.

[The role of phosphoinositides, protein kinase C and protein kinase A in the K+ regulatory signal transduction in human adrenocortical cells].

The messenger mechanisms mediating K+ regulatory signals in human adrenocorticocytes were studied. It was shown that potassium ions initiated decay of polyphosphoinositides to inositolphosphates and obviously diacylglycerol. The latter compounds activate protein kinase C as affected by different agonists. Using western blotting method we showed translocation of PKCalpha from cytosol to membranes after adrenal tissue preincubation in the medium with increased K+ content (8.5 mM). Translocation means activation of the enzyme. Activity of PKC increased in the microsomal fraction and did not change in cytosol. Increased concentration of K+ in the incubation medium also activates protein kinase A, although to a lesser extent compared to PKC. Unlike PKC activity of PKA was changed in cytosol as well. The possibility of involvement of several messenger systems in K+ signal transduction in human adrenocortical cells as well as the hypothesis on cross-talk between messenger mechanisms for main physiological agonists controlling aldosterone biosynthesis in the adrenals are discussed.

[ACTH activates in vitro protein kinases A and C in human adrenal cortex].

The activation of protein kinases A and C by corticotropin in the human adrenal cortex was analyzed. ACTH influence in vitro significantly increase protein kinase A activity, total protein kinase C activity in the membrane fraction of adrenocorticocytes and 11-hydroxycorticosteroids production of adrenal cortex tissue. Either cAMP-dependent protein kinase A or protein kinase C are included to postreceptoral messenger mechanism, that mediates ACTH action in adrenocortical cells.

[The study of iodine sufficiency in children of Kiev]. / Vyvchennia iodnoï zabezpechenosti ditei m. Kyieva.

A total of 576 pupils in different age groups of a school in Kiev were examined. Determined in the above pupils by palpation and echography were sizes of the thyroid gland, urine excretion of iodine. Thyroid enlargement was identified in nearly 30 percent of the examined children, nodal pathology of the gland was in 0.52 percent. It was only in 34 percent of the examinees that the level of iodine excretion was more than 100 mcgrm/l, i.e. did not deviate from the norm; 5.3% of children had ioduria indices < 20 mcgrm/l, which observation suggested a severe iodine deficiency. In the major proportion of children the level of ioduria ranged from 50 up to 100 mcgrm/l, the median value for the group of children as a whole came up to 62.9 mcgrm/l. According to results of the studies made on goiter rates among children and on the level of urine excretion of iodine, the population are in want of bodily iodine, and Kiev falls in to a zone of mild iodine deficiency.

[Use of vilosen in the treatment of radiation damage of the immune system]. / Vilozen iak zasib korektsiï radiatsiinykh urazhen' imunnoï systemy.

The possibility of vilosen usage for the immune system damage liquidation was studied. Rats obtained discrete rentgen irradiation during 1 month in the total dose of 4 Gr. Mice obtained internal 131I irradiation in a dose of 9.25 kBk/g. It was established that thymus and spleen masses, quantity of their cells, blood leukocytes and antibody production decreased by as external and internal irradiation. Irradiated animals treated with vilosen restored their immune system functional state partly or completely. The preparation was assumed to be used for the correction of immune system radiation damage.

[The reactions of the immune system under the influence of a nonpeptide splenic factor]. / Reaktsiï imunnoï systemy pid vplyvom nepeptydnoho chynnyka selezinky.

The influence of low-molecular nonpeptide preparation splenosid obtained from bovine spleen on the cytotoxic activity of natural killers, reaction of complement binding and the immune responsibility was investigated. It was shown that even single treatment of rats with splenosid in a dose of 300 mkg/100 g in 48 hours stimulates the cytolytic activity of splenocytes on target cells (sheep erythrocytes). The same results were obtained in vitro. The treatment of guinea pigs with splenosid reduces the complement activity in serum. This preparation depresses the splenocyte immunoreactivity in sensibilized mice in a dose-dependent manner. The data obtained complete our knowledges about biological activity of splenosid.