[The combination of classical tests and methods of chromatography for laboratory diagnostics of cushing's syndrome of different genesis.] / Сочетание ÐºÐ»Ð°ÑÑÐ¸Ñ‡ÐµÑÐºÐ¸Ñ Ñ‚ÐµÑÑ‚Ð¾Ð² и методов Ñ Ñ€Ð¾Ð¼Ð°Ñ‚Ð¾Ð³Ñ€Ð°Ñ„Ð¸Ð¸ в лабораторной диагностике синдрома Кушинга различного генеза.

Immunochemical methods of analysis are traditionally used for diagnosis of various forms of Cushing's syndrome (CS). In the presence of boundary values of hormonal parameters, doubtful situations, a combination of changes both in pituitary and in adrenal glands, it is useful to determine additional differential diagnostic criteria for the diagnosis of various forms of CS. Urinary steroid profiles (USP) were analyzed by gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) for 38 females with adrenal Cushing's syndrome (CSA), 42 females with pituitary CS (CSP) and 25 healthy females (control group). An increase of free cortisol/free cortisone ratio in the urine (UFF/UFE) for CSP patients in comparison of CSA was obtained by HPLC method. Decreased urinary excretion of UFF and UFE by more than 60% after the 8 mg dexamethasone suppression test had 100% sensitivity and specificity of more than 90% for the diagnosis of CSP. GC-MS method in patients with CSA and CSG revealed the peculiarities of androgens, progestins and glucocorticoids metabolism which leaded to obtain specific USP for CS of different genesis. Increased urinary excretions of dehydroepiandrosterone and its metabolites, metabolites of androstenedione, the ratio of sum of cortisol and cortisone tetrahydrometabolites to tetrahydro-11-deoxycortisol (more then 36) in CSP patients compared with CSA are additional signs for differential diagnosis of these diseases. The combination of classical tests and USP obtained by HPLC and GC-MS methods increased the sensitivity and specificity of differential diagnosis of CSA and CSP.

Gas Chromatography-Mass Spectrometry Analysis of Urinary Steroid Metabolomics for Detection of Early Signs of Adrenal Neoplasm Malignancy in Patients with Cushing's Syndrome.

The metabolomics of urinary steroids was studied by gas chromatography-mass spectrometry in 25 patients with Cushing's syndrome without malignant potential and in 12 patients with malignant potential of adrenal neoplasms (Weiss score 1-3). Patients with adrenocortical adenoma (N=24) constituted the control group. In patients with Cushing's syndrome and malignant potential, increased urinary excretion of 16-oxo-androstendiol, tetrahydro-11-deoxycortisol, and 16-hydroxypregnendiol, which had 100% specificity and sensitivity >90% for the diagnosis of malignant potential. Additionally, non-classical 5-ene-pregnenes (16-OHpregnenolone, 21-OH-pregnenolone, 3ß,16,20-pregnentriol, and 3ß,17,20-pregnentriol) were identified. The revealed changes in the metabolomics of steroids can be early signs of malignant potential in patients with Cushing's syndrome. In patients with malignant potential, three signs of reduced activity of 11ß-hydroxysteroid dehydrogenase type 2 were detected and in patients without malignant potential, one sign was found. In patients with and without malignant potential, three signs increased activity of 5ß-reductase were found.

Different Types of Urinary Steroid Profiling Obtained by High-Performance Liquid Chromatography and Gas Chromatography-Mass Spectrometry in Patients with Adrenocortical Carcinoma.

Urinary steroid profiling (USP) was studied using high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) methods in 108 patients with adrenocortical adenoma (ACA) and in 31 patients with adrenocortical carcinoma (ACC). Thirteen ACC and Cushing's syndrome (ACC-CS) patients had two types of USP as well as 18 ACC patients without hypercortisolism. These four types differed by androgen and glucocorticoid secretion of the adrenal cortex. Fifteen main ACC features were observed by GC-MS. Urinary excretion of dehydroepiandrosterone (DHEA) was increased in 67.7 % of ACC patients and tetrahydro-11-deoxycortisol (THS) in 74.2 %. By combination of the following parameters: THS >900 µg/24 h and/or DHEA >1500 µg/24 h with ratios of 3α,16,20-pregnentriol/3ß,16,20-pregnentriol (3α,16,20dP3/3ß,16,20dP3) less than 6.0 and 3α,17,20dP3/3ß,17,20dP3 less than 9.0 and the detection of "non-classical" 5-en-pregnens, not found in ACA and healthy persons, 100 % sensitivity and specificity of ACC and ACA differential diagnosis were achieved. Features of 21-hydroxylase and 11ß-hydroxylase deficiency were observed by GC-MS in 32.2 and 61.3 % of the ACC patients, respectively. Additional features for ACC-CS diagnostic were increased urinary excretion of 6ß-hydroxycortisol, 18-hydroxycorticosterone, the sum (UFF + UFE) obtained by HPLC, tetrahydrocorticosterone, and the sum (THF + THE + allo-THF) obtained by GC-MS.

[Specific features of diagnosis of hormonal insufficiency in patients with tumors of the adrenal cortex].

An examination of 145 patients with incidentalomas and 195 women with virale syndrome has shown that 20% of patients with incidentalomas and 23.1% of patients with steroidogenesis have disorders of the adrenal steroidogenesis characteristic of the obliterated form of congenital hyperplasia of the adrenal cortex (CHAC) with a defect of 11beta-hydroxylase on the basis of the following biochemical criteria: the elevation in blood of the basal levels of 11-desoxycortisole and 11beta-desoxycorticosterone, decreased excretion of free cortisole with urine, lower indices of hydrocortisole/cortisone in blood and free cortisole/free cortisone in urine, in the test with corticotropin - elevation in blood of the level of 11-desoxycortisole and 11-desoxycorticosterone, decreased relationships cortisole/11-desoxycortisole and cortisole/cortisone and lower growth of the levels of corticosterone and cortisole. The data obtained suggest that long-standing obliterated form of CHAC with a defect of 11beta-hydroxylase might be the cause of the formation of certain incidentalomas.

[Diagnostic value of high performance liquid chromatography of corticosteroids in diseases of the pituitary system].

A hundred and ninety-one patients with different adrenal cortical tumors were studied to assess the diagnostic value of HPLC of corticosteroids in the biological fluids. HPLC of corticosteroids in the biological fluids revealed the signs of the hormonal activity of incidentalomas, organic endogenous hypercorticism, and mass malignancy. Some patients with adrenal cortical incidentalomas (15.5%) were found to have impaired steroid formation that is typical of 11ß-OH- or 17-OH-hydroxylase deficiencies. The findings allow one to diagnose adrenal disorders at the early stages of disease and to define indications for surgical treatment.

[The specific features of adrenal steroidogenesis in patients with congenital adrenal cortical dysfunction due to 21-hydroxylase deficiency].

Forty-two patients with classical forms of congenital adrenal cortical dysfunction (CACD) (22 patients with salt-loosing form and 20 patients with simple virile form) caused by 21-hydroxylase deficiency and 29 female with an obscure form of the disease were examined. The patients with classical CACD received therapy with gluco- and mineralocorticoids. Radioimmunoassay and enzyme immunoassay were employed to estimate the levels of 17-OH-progesterone and the activities of plasma renin and serum corticotropin, aldosterone, dehydroepiandrosterone, dehydroepiandrosterone sulfate, and androstenedione. High performance liquid chromatography (HPLC) was used to measure the blood levels ofcortisol, cortisone, corticosterone (B), 11-deoxycorticosterone (DOC), and 11-deoxycortisol (S). There were increases in the levels of В and DOC in patients with the simple form at the decompensation stage and in those of DOC in patients with the simple form at the compensation stage, which is indicative of activation of the renin-angiotensin system. There were also rises in В and DOC in 35.8 and 33.0% of the patients with obscure CACD, respectively, as well as a significant elevation of the content of S. A study of the profiles of corticosteroids by HPLC in patients with 21-hydroxylase deficiency-related CACD is an additional method for assessing the compensation of the disease in patients with classical CACD and for diagnosing the obscure form of the disease.

[Minidiab test to assess the functional state of pancreatic beta-cells in the children of diabetics]. / Proba s minidiabom v otsenke funktsional'nogo sostoianiia beta-kletok podzheludochnoi zhelezy u potomkov bol'nykh sakharnym diabetom.

The levels of glucose, insulin, and C-peptide in the blood serum were measured in 38 subjects with normal and impaired glucose tolerance whose parents suffered from insulin-dependent and noninsulin-dependent diabetes mellitus (IDDM and NIDDM, respectively) and in 12 normal subjects without hereditary aggravation for diabetes mellitus in order to specify the peculiarities of development of diabetes mellitus of various types. Reliably increased levels of glucose, insulin, and C-peptide on an empty stomach and absence of adequate secretion of insulin and C-peptide in response to stimulation with 5 mg of minidiab, expressed by a later and less manifest release of insulin and C-peptide, were observed in the test group, in contrast to healthy controls. The detected changes augment with the progress of carbohydrate metabolism disorders, being more marked in the subjects whose parents suffered from IDDM. The findings permit a conclusion that function of the insular system is changed during early disorders of carbohydrate metabolism in subjects whose parents suffered from both forms of diabetes mellitus. Minidiab test is recommended to specify the function of the pancreatic insular system.

[Immunologic indices in Itsenko-Cushing disease and syndrome]. / Immunologicheskie pokazateli pri bolezni i sindrome Itsenko-Kushinga.

Examination of 13 untreated patients with Itsenko--Cushing disease and syndrome showed an increase in total Ig as well as IgA, IgM and IgG, a decrease in the number of O-cells, and the absence of significant change in in the blood content of T-lymphocytes. A decrease in the amount of B-lymphocytes was noted in this group of patients with GTT of diabetic type or mild diabetes mellitus. Repeated examination of 7 of these patients 1-1.5 mos. after the resection of one adrenal gland showed that the content of O-cells had returned to normal, tension of the system of humoral immunity was on a decrease and combined with a decrease in the excretion of 17-OCS fractions though they were still above normal. Hyperlymphocytosis with a high level of T- and B-lymphocytes and the normal content of O-cells was noted in 11 patients after bilateral adrenalectomy for Itsenko--Cushing disease receiving maintenance doses of cortisone.

[Role of insulin and its antagonists in the pathogenesis of diabetes mellitus and its complications]. / Rol' antagonistov i insulina v patogeneze sakharnogo diabeta i ego oslozhnenii.

The content of insulin antagonists (growth hormone, cortisol, catecholamine, DOPA, free fatty acids) was studied in 40 women suffering from diabetes mellitus, including 12 insulin-sensitive and 15 insulin-resistant, and 13 women with freshly diagnosed untreated diabetes. The greatest content of all the insulin antagonists was revealed in the insulin-sensitive patients during the decompensation period. It is assumed that in this period, diabetogenic actions of insulin antagonists create the most favourable conditions for the formation of insulin antagonists create the most favourable conditions for the formation of insulin resistance; this resistance is then maintained by the increased antibody titre due to the elevation of therapeutic insulin doses and lengthy persistence of the high growth hormone level in the insulin-resistant patients. Besides, the high growth hormone content promotes angioretinopathy development. A conclusion is drawn that the content of growth hormone, cortisol, catecholamines, DOPA, or free fatty acids does not serve as the primary factor in the pathogenesis of diabetes mellitus, since at any of the phases of the disease the level of each of them failed to differ from control.