Development of New Liquid Chromatographic Method for Mitotane and Its Metabolites Determination in Human Plasma Employing Design of Experiments Methodology

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 C-18 with pre-column Restek Ultra Aqua C-18 operating at temperature 35 degrees C; mixture of acetonitrile and 0.5% formic acid as mobile phase with 1.2mL min(-1) flow rate and detection at 230nm. 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.This is peer-reviewed version of the following article: Jancic-Stojanovic, B.; Vemić, S.; Elezović, V.; Petrović, A.; Sinadinović, Z.; Ivanović, D.; Damjanović, S.; Miljković, B. Development of New Liquid Chromatographic Method for Mitotane and Its Metabolites Determination in Human Plasma Employing Design of Experiments Methodology. J. Liq. Chromatogr. Relat. Technol. 2015, 38 (14), 1371–1378. [https://doi.org/10.1080/10826076.2015.1057645

The utility of Tc-99m-EDDA/HYNIC-TOC scintigraphy for assessment of lung lesions in patients with neuroendocrine tumors

Our aim was to assess clinical utility of Tc-99m-EDDA/HYNIC-TOC scintigraphy for evaluation of lung lesions in patients with neuroendocrine tumors (NETs). Single photon emission computed tomography (SPECT) of the thorax and whole body scintigraphy were performed in 34 patients using Tc-99m-EDDA/HYNIC-TOC. Visual assessment was complemented by semiquantitative evaluation based on tumor to non-tumor (TINT) ratio. Clinical, laboratory, and histological findings served as the standard for comparison. Enhanced tracer uptake was observed on both SPECT and whole body scintigraphy in 29 of 34 patients (88% sensitivity). TINT ratios were significantly higher on SPECT than whole body images (2.96 +/- 1.07 vs. 1.70 +/- 0.43, p LT 0.01) and did not correlate with NET proliferation index Ki-67 (r= - 0.36, p=0.27). Conclusion: Tc-99m-EDDA/HYNIC-TOC scintigraphy is useful for evaluation of NET tissue in the lungs. SPECT provides better visualization of lung lesions than whole body scintigraphy. The intensity of tracer uptake, however, does not relate to the proliferation rate of NETs. Tc-99m-EDDA/HYNIC-TOC scintigraphy may be helpful for selecting and monitoring treatment options, particularly when radiolabeled somatostatin analogue therapy becomes available

The role of personality and traumatic events in cortisol levels - Where does PTSD fit in?

Background: Studies of cortisol in post-traumatic stress disorder (PTSD) have yielded mixed results. We hypothesize that personality traits and traumatic experiences could be the confounders of cortisol measures and disease symptoms. Method: This study was a part of a broader project in which simultaneous psychological and biological investigations were carried out in hospital conditions on 400 male participants categorized by four groups: (A) 133 with current PTSD, (B) 66 with lifetime PTSD, (C) 102 trauma controls, and (D) 99 healthy controls (matched by age and education). Cortisol and ACTH were measured in blood samples taken hourly from 22:00 h to 09:00 h, with an additional sample at 07:30 h (resting state and morning rise). The next night, dexamethasone (0.5 mg) suppression test was performed. Results: No significant differences in basal cortisol and ACTH were found between study groups. The trait Conscientiousness, negatively modulated by Extraversion (assessed by NEO Personality Inventory-Revised) was found to correlate with cortisol (but not with ACTH). Group differences are found on suppression. Structural equation modeling shows excellent fit only when the paths (influences) from Conscientiousness to basal cortisol and from traumatic events to suppression are present. The paths connecting suppression and PTSD symptoms do not contribute. Conclusions: Two sources of differences of hypothalamo-pituitary-adrenocortical axis functioning are implied, both only indirectly connected to PTSD. It seems that basal cortisol secretion is associated more tightly with personality (introvertively modulated Conscientiousness), while the regulation by glucocorticoid receptor system is sensitized by repeated traumatic situations. (c) 2011 Elsevier Ltd. All rights reserved

Is there a biological difference between trauma-related depression and PTSD? DST says NO

The use of the low-dose dexamethasone suppression test (DST) as a potentially discriminative marker between post-traumatic stress disorder (PTSD) and depression is still under discussion. In order to compare the influence of these psychopathologies on the DST results, we examined suppression in war-traumatized subjects with one or both of these disorders, as well as in healthy controls. Based on our previous findings, we hypothesized that subjects with any disorder would exhibit higher dexamethasone suppression than healthy controls due to traumatic experiences. This study was a part of a broader project in which simultaneous psychological and biological investigations were carried out in hospital conditions on 399 mate participants: 57 with PTSD, 28 with depression, 76 with PTSD + depression, and 238 healthy controls. Cortisol was measured in blood samples taken at 0900 h before and after administering 0.5 mg of dexamethasone (at 2300 h). Group means standard deviation of cortisol suppression were: 79.4 +/- 18.5 in the PTSD group, 80.8 +/- 11.6 in the depression group, 77.5 +/- 24.6 in the group with PTSD+depression, and 66.8 +/- 34.6 in healthy controls. The first three groups suppressed significantly more than the fourth. When the number of traumas was introduced as a covariate, the differences disappeared. The hypothesis was confirmed: in respect to DST, the examined trauma-related psychopathologies showed the same pattern: hypersuppression, due to multiple traumatic experiences. (C) 2012 Elsevier Ltd. All rights reserved

Is there a biological difference between trauma-related depression and PTSD? DST says 'NO'

The use of the low-dose dexamethasone suppression test (DST) as a potentially discriminative marker between post-traumatic stress disorder (PTSD) and depression is still under discussion. In order to compare the influence of these psychopathologies on the DST results, we examined suppression in war-traumatized subjects with one or both of these disorders, as well as in healthy controls. Based on our previous findings, we hypothesized that subjects with any disorder would exhibit higher dexamethasone suppression than healthy controls due to traumatic experiences. This study was a part of a broader project in which simultaneous psychological and biological investigations were carried out in hospital conditions on 399 mate participants: 57 with PTSD, 28 with depression, 76 with PTSD + depression, and 238 healthy controls. Cortisol was measured in blood samples taken at 0900 h before and after administering 0.5 mg of dexamethasone (at 2300 h). Group means standard deviation of cortisol suppression were: 79.4 +/- 18.5 in the PTSD group, 80.8 +/- 11.6 in the depression group, 77.5 +/- 24.6 in the group with PTSD+depression, and 66.8 +/- 34.6 in healthy controls. The first three groups suppressed significantly more than the fourth. When the number of traumas was introduced as a covariate, the differences disappeared. The hypothesis was confirmed: in respect to DST, the examined trauma-related psychopathologies showed the same pattern: hypersuppression, due to multiple traumatic experiences. (C) 2012 Elsevier Ltd. All rights reserved.European Commission [INCO-CT-2004-509213]; Ministry of Science, Serbia [179018, 41009

The role of personality and traumatic events in cortisol levels - Where does PTSD fit in?

Background: Studies of cortisol in post-traumatic stress disorder (PTSD) have yielded mixed results. We hypothesize that personality traits and traumatic experiences could be the confounders of cortisol measures and disease symptoms. Method: This study was a part of a broader project in which simultaneous psychological and biological investigations were carried out in hospital conditions on 400 male participants categorized by four groups: (A) 133 with current PTSD, (B) 66 with lifetime PTSD, (C) 102 trauma controls, and (D) 99 healthy controls (matched by age and education). Cortisol and ACTH were measured in blood samples taken hourly from 22:00 h to 09:00 h, with an additional sample at 07:30 h (resting state and morning rise). The next night, dexamethasone (0.5 mg) suppression test was performed. Results: No significant differences in basal cortisol and ACTH were found between study groups. The trait Conscientiousness, negatively modulated by Extraversion (assessed by NEO Personality Inventory-Revised) was found to correlate with cortisol (but not with ACTH). Group differences are found on suppression. Structural equation modeling shows excellent fit only when the paths (influences) from Conscientiousness to basal cortisol and from traumatic events to suppression are present. The paths connecting suppression and PTSD symptoms do not contribute. Conclusions: Two sources of differences of hypothalamo-pituitary-adrenocortical axis functioning are implied, both only indirectly connected to PTSD. It seems that basal cortisol secretion is associated more tightly with personality (introvertively modulated Conscientiousness), while the regulation by glucocorticoid receptor system is sensitized by repeated traumatic situations. (c) 2011 Elsevier Ltd. All rights reserved

Is there a biological difference between trauma-related depression and PTSD? DST says 'NO'

The use of the low-dose dexamethasone suppression test (DST) as a potentially discriminative marker between post-traumatic stress disorder (PTSD) and depression is still under discussion. In order to compare the influence of these psychopathologies on the DST results, we examined suppression in war-traumatized subjects with one or both of these disorders, as well as in healthy controls. Based on our previous findings, we hypothesized that subjects with any disorder would exhibit higher dexamethasone suppression than healthy controls due to traumatic experiences. This study was a part of a broader project in which simultaneous psychological and biological investigations were carried out in hospital conditions on 399 mate participants: 57 with PTSD, 28 with depression, 76 with PTSD + depression, and 238 healthy controls. Cortisol was measured in blood samples taken at 0900 h before and after administering 0.5 mg of dexamethasone (at 2300 h). Group means standard deviation of cortisol suppression were: 79.4 +/- 18.5 in the PTSD group, 80.8 +/- 11.6 in the depression group, 77.5 +/- 24.6 in the group with PTSD+depression, and 66.8 +/- 34.6 in healthy controls. The first three groups suppressed significantly more than the fourth. When the number of traumas was introduced as a covariate, the differences disappeared. The hypothesis was confirmed: in respect to DST, the examined trauma-related psychopathologies showed the same pattern: hypersuppression, due to multiple traumatic experiences. (C) 2012 Elsevier Ltd. All rights reserved.European Commission [INCO-CT-2004-509213]; Ministry of Science, Serbia [179018, 41009

Pheochromocytoma in von Hippel-Lindau disease

A 70-year old female was admitted to the hospital because of hypertension increased sweating and weight loss. The hypertension was sustained. Five months before admission CT scan of the abdomen had revealed a well-defined right adrenal mass together with left kidney tumor. A magnetic resonance imaging of the abdomen confirmed the presence of the right adrenal and left kidney masses, but also showed another tumor in the pancreas between the body and the tail. Urinary 24-hour noradrenaline was grossly elevated and confirmed the diagnosis of pheochromocytoma. 131I-metaiodobenzylgvanidine (MIBG) scintiscan showed increased MIBG uptake in the right adrenal gland. After pre-treatment with phenoxybenzamine 30 mg daily, the patient was operated, and the right adrenalectomy was done. Histopathological examination revealed encapsulated adrenal pheochromocytoma without infiltrative characteristics and lymph node metastasis. After the operation hypertension was controlled easily with amlodipine. The patient was discharged for recovery. Ulteriorly, SSCP (single strand conformational polymorphism) method detected a point mutation in the third exon of the VHL (von Hippel-Linday) gene. It was decided to follow up the patient with the von Hippel-Lindau disease, while waiting for the results of the sequence analysis to confirm that the found mutation is not associated with renal cancer

Hypothalamic-Pituitary-Adrenocortical Axis Hypersensitivity and Glucocorticoid Receptor Expression and Function in Women with Polycystic Ovary Syndrome

Introduction: Molecular mechanisms underlying pathophysiology of polycystic ovary syndrome (PCOS), especially those related to cortisol signaling, are poorly understood. We hypothesized that modulation of glucocorticoid receptor (GR) expression and function, may underlie possible PCOS-related impairment of feedback inhibition of hypothalamic-pituitary-adrenocortical (HPA) axis activity and thus contribute to increased adrenal androgen production in women with PCOS. Materials and Methods: 24 normal-weight and 31 obese women with PCOS were compared to 25 normal-weight controls. Fasting blood samples were collected for measurements of serum concentrations of dehydroepiandrosterone sulfate, testosterone, sex hormone-binding globulin, insulin, basal cortisol and cortisol after oral administration of 0.5 mg dexamethasone. Concentrations of GR mRNA, GR protein, mineralocorticoid receptor (MR) protein and heat shock proteins (Hsps), as well as the number of GR per cell (B(max)) and its equilibrium dissociation constant (K(D)) were measured in isolated peripheral blood mononuclear cells. Results: An increase in HPA axis sensitivity to dexamethasone, an elevation of the GR protein concentration, and unaltered receptor functional status were found in both normal-weight and obese women with PCOS vs. healthy controls. Lymphocyte MR, Hsp90 and Hsp70 concentrations, and MR/GR ratio were similar in all groups. Correlation between B(max) and K(D) was weaker in the group of obese women with PCOS than in the other 2 groups. Conclusions: The results did not confirm the initial hypothesis, but imply that PCOS is associated with increased GR protein concentration and HPA axis sensitivity to dexamethasone.Ministry of Education and Science, Serbia [41009

Exercise capacity is not impaired after acute alcohol ingestion: a pilot study

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 1.5g/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 10min 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 parts per thousand (P lt 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 +/- 543ml/min, ns; peak workload=269 +/- 53 vs. 249 +/- 40W, ns; test duration 13.7 +/- 2.2 vs. 13.3 +/- 1.7min, 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