[Show abstract][Hide abstract] ABSTRACT: A simple and reliable new HPLC method with UV detection has been developed and validated for simultaneous determination of mitotane and its two metabolites DDA and DDE. Method development was carried out utilizing systematical approach of the Design of Experiments (DoE) methodology. For estimation of factors influence on selected chromatographic responses and definition of the optimal chromatographic conditions, Box-Behnken experimental design was applied. The defined optimal separation conditions were: column Restek Ultra Aqua C18 with pre-column Restek Ultra Aqua C18 operating at temperature 35 °C; mixture of acetonitrile and 0.5% formic acid as mobile phase with 1.2 mL min−1 flow rate and detection at 230 nm. As sample preparation method, liquid-liquid extraction was chosen. Method was fully validated and LOQ and LOD were experimentally determined. Finally, method was successfully applied for determination of mitotane and its metabolites in plasma samples of patients with adrenocortical carcinoma.
Journal of Liquid Chromatography & Related Technologies 06/2015; 38(14):150611134831005. DOI:10.1080/10826076.2015.1057645 · 0.61 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Objective: Certain Bcll glucocorticoid receptor gene polymorphism is found to be linked with tissue specific glucocorticoid sensitivity and associated with metabolic syndrome. The aim of this study was to evaluate association of Bcll polymorphism with the occurrence of side effects of the treatment with intravenous methylprednisolone (iv MP) in patients with Graves' orbitopathy (GO). Glucocorticoids (GC) are used in the treatment of a broad (wide) spectrum of diseases in the dosage and duration (of) which often leads to moderate to severe complications. Although there are frequent side effects, weight gain, hypertension, dyslipidemia, impaired glycemic control parameters are not predictably present. Design and method: The study group included 22 subjects (17 female and 5 male) treated with two doses of 500 mg iv MP during three days, followed by oral prednisone 40 mg/d tapered to 10 mg/d during four weeks. Six courses of therapy were performed. Results: Nine patients had no glucocorticoid gene receptor polymorphism, 11 were heterozygous for Bcll and 2 patients were homozygous. All patients had a good response to the treatment of GO. Homozygous patients were slightly overweight with normal metabolic profile, but during the treatment cholesterol and blood pressure increased. They also tended to develop hypokalemia. Three of nine patients (1 obese, 2 overweight, 6 with normal weight) with no polymorphism had glucose intolerance, mild arterial hypertension and increased cholesterol before the treatment and they were stable during the treatment. Moderate hypertension in two subjects were also stable with no additional therapy. Four of eleven heterozygous patients (2 obese, 2 overweight, 7 with normal weight) had glucose intolerance and increased cholesterol, but during the treatment they normalized metabolic profiles by balanced nutrition. Two of three patients with well controlled moderate hypertension observed worsening of blood pressure. Conclusions: In conclusion, there was no significant difference in distribution of arterial hypertension before the treatment by glucocorticoids between groups with and without Bcll polymorphism. In this small group of patients Bcll polymorphism was not useful in prediction of blood pressure or other metabolic parameters response during the treatment. Copyright
Journal of Hypertension 06/2015; 33:e511-e512. DOI:10.1097/01.hjh.0000469003.77341.f1 · 4.72 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Background
The hypothalamo-pituitary-adrenocortical (HPA) axis self-regulation is achieved via cortisol binding to mineralocorticoid (MR) and glucocorticoid receptors (GR). It is often disturbed in mental disorders, particularly in those where traumatic stress has been implicated, such as posttraumatic stress disorder and depression. Although dexamethasone suppression test (DST) is often used as diagnostic aid, the findings still vary. In search of the factors influencing the DST outcome, we examined the glucocorticoid receptor (GR) gene BclI polymorphism.
A total of 229 male subjects were classified into three BclI groups: two groups with homozygous carriers (of the G allele, N=108, and of the C allele, N=26), and one with heterozygous carriers (N=95). Multiple hierarchical linear regression analysis was done, where the dependent variable was the dexamethasone-induced cortisol suppression, and predictors included receptor variables. The interactions of the count of ‘G׳s with the predictors were introduced to single out the effects of the G allele.
The means of all studied variables, including suppression, are statistically the same in the three groups. However, the mechanism of suppression involves MRs only in the G allele carriers.
The subjects were selected by criteria suited for the aim of the large project whose part is this study, hence the relatively small number of CC carriers. Also, we did not assess MR functional properties that would probably sharpen the results.
Our finding that MRs participate in cortisol suppression in the G allele carriers suggests that research aimed at refining HPA axis-based therapy might require its adjustment for such patients.
[Show abstract][Hide abstract] ABSTRACT: The usage of alcohol is widespread, but the effects of acute alcohol ingestion on exercise performance and the stress hormone axis are not fully elucidated.We studied 10 healthy white men, nonhabitual drinkers, by Doppler echocardiography at rest, spirometry, and maximal cardiopulmonary exercise test (CPET) in two visits (2-4 days in between), one after administration of 1.5 g/kg ethanol (whisky) diluted at 15% in water, and the other after administration of an equivalent volume of water. Plasma levels of NT-pro-BNP, cortisol, and adrenocorticotropic hormone (ACTH) were also measured 10 min before the test, at maximal effort and at the third minute of recovery. Ethanol concentration was measured from resting blood samples by gas chromatography and it increased from 0.00 ± 0.00 to 1.25 ± 0.54‰ (P < 0.001). Basal echocardiographic and spirometric parameters were normal and remained so after acute alcohol intake, whereas ACTH, cortisol, and NT-pro-BNP nonsignificantly increased in all phases of the test. CPET data suggested a trend toward a slight reduction of exercise performance (peak VO2 = 3008 ± 638 vs. 2900 ± 543 ml/min, ns; peak workload = 269 ± 53 vs. 249 ± 40 W, ns; test duration 13.7 ± 2.2 vs. 13.3 ± 1.7 min, ns; VE/VCO2 22.1 ± 1.4 vs. 23.3 ± 2.9, ns). Ventilatory equivalent for carbon dioxide at rest was higher after alcohol intake (28 ± 2.5 vs. 30.4 ± 3.2, P = 0.039) and maximal respiratory exchange ratio was lower after alcohol intake (1.17 ± 0.02 vs. 1.14 ± 0.04, P = 0.04). In conclusion, we showed that acute alcohol intake in healthy white men is associated with a nonsignificant exercise performance reduction and stress hormone stimulation, with an unchanged exercise metabolism.
Journal of Cardiovascular Medicine 07/2014; Publish Ahead of Print. DOI:10.2459/JCM.0000000000000151 · 1.51 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to evaluate the predictive value of adrenocorticotropic hormone (ACTH), cortisol and ACTH receptor polymorphism (ACTHRP) for left ventricular (LV) remodeling. Thirty-six elite male athletes, as chronic stress adaptation models, and twenty sedentary age and sex-mached subjects emabarked on standard and tissue Doppler echocardiography to assess cardiac parameters at rest. They performed maximal cardiopulmonary test, which was used as an acute stress model. ACTH and cortisol were measured at rest (10 min before test), at beginning, at maximal effort, at 3rd min of recovery, using radioimmunometric and radioimmunoassey techniques, respectively. Promoter region of ACTHR gene (18p11.2) was analysed from blood samples using reverse polymerisation reaction with the analysis of restriction fragment lenght polimorphisam by SacI restriction enzyme. Normal genotype was CTC/CTC, heterozygot for ACTHRP CTC/CCC and homozygot CCC/CCC. In all participants, ACTH and cortisol increased during acute stress, whereas in recovery ACTH increased and cortisol remained unchanged. 49/56 examiners manifested CTC/CTC, 7/56 CTC/CCC and 0/56 CCC/CCC. There was no difference in ACTHRP frequency between groups (χ2(1) = 0,178, p = 0.67). LV mass (LVM) and LV end-diastolic volume (LVVd) were higher in athletes than in controls (p < 0.01) and lower in CTC/CTC than in CTC/CCC genotype (219.43 ± 46.59(SD)g vs. 276.34 ± 48.86(SD)g, p = 0.004; 141.24 ± 24.46(SD)ml vs. 175.29 ± 37.07(SD)ml, p = 0.002; respectively). In all participants, predictors of LVM and LVVd were ACTH at rest (B = -1.00,-0.44; β=-0.30;-0.31; p = 0.026,0.012, respectively) and ACTHRP (B = 56.63,34; β=0.37,0.40; p = 0.003,0.001, respectively). These results demonstrate that ACTH and ACTHRP strongly predict cardiac morphology suggesting possible regulatory role of stress system activity and sensitivity in cardiac remodeling.
[Show abstract][Hide abstract] ABSTRACT: The multiple endocrine neoplasia type 2A (MEN 2A) syndrome, comprising medullary thyroid carcinoma (MTC), pheochromocytoma and primary hyperparathyroidism (PHPT) is most frequently caused by codon 634 activating mutations of the RET (rearranged during transfection) proto-oncogene on chromosome 10. For this codon-mutation carriers, earlier thyroidectomy (before the age of 5 years) would be advantageous in limiting the potential for the development of MTC as well as parathyroid adenomas.
This is a case report of 3-year-old boy from the MEN 2A family (the boy's father and grandmother and paternal aunt) in which cysteine substitutes for phenylalanine at codon 634 in exon 11 of the RET proto-oncogene, who underwent thyroidectomy solely on the basis of genetic information. A boy had no thyromegaly, thyroidal irregularities or lymphadenopathy and no abnormality on the neck ultrasound examination. The pathology finding of thyroid gland was negative for MTC. Two years after total thyroidectomy, 5-year-old boy is healthy with permanent thyroxine replacement. His serum calcitonin level is < 2 pg/ml (normal < 13 pg/ml), has normal serum calcium and parathyroid hormone levels and negative urinary catecholamines. Long-term follow-up of this patient is required to determine whether very early thyroidectomy improves the long-term outcome of PHPT.
Children with familial antecedents of MEN 2A should be genetically studied for the purpose of determining the risk of MTC and assessing the possibilities of making prophylactic thyroidectomy before the age of 5 years.
Srpski arhiv za celokupno lekarstvo 03/2014; 142(1-2):72-74. DOI:10.2298/SARH1402072J · 0.23 Impact Factor