ACTA ENDOCRINOLOGICA (BUC)

Background. Methotrexate (MTX) is used in the treatment of neoplastic disorders. MTX action in non-neoplastic diseases still remains obscure. Female reproductive cells are fast proliferating like cancer cells. Hence, it is important to\r\nidentify markers affected by MTX in the reproductive tract.\r\nAim. Investigate MTX effect on energy metabolism marker glycogen and the protective role of leucovorin (LCN) and\r\nwithdrawal of MTX treatment in the ovary, oviduct, uterus, cervix and vagina of rats.\r\nAnimals and Methods. Rats with regular oestrous cycle were randomly divided into five groups (n=6) as follows: Control, MTX LD (low dose), MTX HD (high dose), MTXHD+LCN (leucovorin), and MTXHD+WD (withdrawal): 20 days withdrawal. Animals were treated once per day intramuscularly for 20 days. Rats were sacrificed on day 21. MTXHD treatment was withdrawn for additional 20 days and\r\nanimals sacrificed on day 41. Ovary, oviduct, uterus, cervix and vagina were used for glycogen analysis.\r\nResults. The present study explored the effect of MTX and LCN on glycogen content in the ovary, oviduct, uterus, cervix and vagina of rats. MTX significantly (P<0.001) inhibited glycogen content in ovary, oviduct, uterus, cervix and vagina of rats, which was dose dependent. LCN\r\nsupplementation and withdrawal of MTX treatment, partially recovered such an effect.

Background and aims. Severe Ovarian Hyperstimulation Syndrome (OHSS) forms with very aggressive clinical evolution are still common, despite prophylactic measures. Besides the Vascular Endothelial Growth Factor (VEGF), there are other angiogenic factors, like Renin-Angiotensin-Aldosterone System (RAS), that might be associated with this disorder. Our study aims to evaluate the role of VEGF and Angiotensin II (ANG II) in the development of early severe OHSS, in high risk patients under prophylactic Cabergoline therapy. Material and Methods. We recruited 192 patients undergoing in vitro fertilization (IVF) procedures with high risk for OHSS development. Out of these, 106 patients with OHSS were enrolled in the study, of which 28 subjects had a severe form of disease (group I), and 78 patients had a mild/ moderate form (group II). We collected blood and follicular fluid from our study participants and determined serum and follicular VEGF and ANG II levels using Enzyme-Linked Immunosorbent Assay (ELISA) technique. Results. Follicular VEGF, ANG II, and serum VEGF levels were significantly higher in group I versus group II. Serum VEGF titers were 645.97 versus 548.62 (p = 0.0008), follicular VEGF titers were 2919.52 versus 1093.68 (p < 0.0001), and follicular ANG II levels were 281.64 versus 65.76 (p < 0.0001). No significant differences have been shown between the two groups for serum ANG II levels. Conclusion. Our study results provide evidence of a OHSS phenotype that is more prone to undergo severe clinical forms of disease, despite treatments with VEGF receptor blockers, and show that ANG II appears to play a major role alongside VEGF, in the development of these severe forms of disease.

Background and Aim. Ischemia-reperfusion (I/R) injury frequently occurs in different situations. Female sex hormones have a protective function. The purpose of this study was to determine the function of female sexual hormones on the gastric damage induced by I/R in male rats. Methods. Forty (40) Wistar rats were randomized into five groups: intact, ischemia- reperfusion (IR), IR + estradiol (1mg/kg), IR + progesterone (16 mg / kg) and IR + combination of estradiol (1mg / kg) and progesterone (16 mg/ kg). Before the onset of ischemia and before reperfusion all treatments were done by intraperitoneal (IP) injection. After animal anesthesia and laparotomy, celiac artery was occluded for 30 minutes and then circulation was established for 24 hours. Results expressed as mean ± SEM and P <0.05 were considered statistically significant. Results. The Glutathione (GSH) concentration significantly decreased after induction of gastric IR (P<0.001). Estradiol (P<0.001) and combined estradiol and progesterone (P<0.001) significantly increased GSH levels. The myeloperoxidase (MPO) concentration significantly increased after induction of gastric IR (P<0.001). Different treatments significantly reduced MPO levels (P<0.001). The gastric acid concentration significantly increased after induction of gastric IR (P<0.001). Treatment with estradiol, progesterone (P<0.05) and combined estradiol and progesterone (P<0.01) significantly reduced gastric acid levels. Superoxide dismutase (SOD) concentration decreased after induction of gastric IR. The SOD levels were not significant. Conclusion. These data suggested that female sexual steroids have a therapeutic effect on gastrointestinal ischemic disorders by reduction of MPO and gastric acid, and increasing gastric GSH & SOD levels following gastric IR.

In some humans, the big and big-big prolactin variants represent the majority of circulating prolactin, considered to be without biological activity. Aims: to establish the clinical expression of macroprolactinemia and the interference with immunodiagnostic tests\r\nin a randomized group of 84 consecutive patients with hyperprolactinemia. IRMA and electrochemiluminescence (Elecsys) were used for PRL assay; gel filtration chromatography (GFC) and protein A precipitation test were used to reveal macroprolactinemia. Results: Macroprolactinemia was found in 16 out of 84 patients (group A), 62 patients had hyperprolactinemia of other causes (group B) and 6 had normal PRL levels and normal GFC (group C). Of 16 patients with macroprolactinemia, 6 showed normal PRL with IRMA and hyperprolactinemia with Elecsys. The difference between the two methods used (&#8710; = PRL determined by Elecsys, -PRL determined by IRMA) correlated with big big PRL level determined by GFC with Elecsys in all patients. The strongest correlation was found in patients with macroprolactinemia (group A, r=0.82, p<0.01) as compared with group B, without macroprolactinemia (r=0.39, p<0.01). Menstrual disorders were expressed, but less frequent in group A versus B (3/15 vs. 28/56, p=0.04), and the appearance of galactorrhea and infertility were not statistically different. Conclusions: In these patients, macroprolactinemia had clinical expression, but weaker than in true hyperprolactinemic patients. It determines high apparent variability of serum PRL level in current commercial assays.

Objective. To characterize serum chemerin levels in obese patients with gestational diabetes mellitus (GDM). Design. Case–control study. Subjects and Methods. Forty seven obese women with newly diagnosed GDM at 24-28 weeks of pregnancy and 32 age, body mass index- and gestational age-matched, normal pregnant women were included. Metabolic patterns and serum chemerin concentrations were measured. Results. Serum chemerin levels were significantly higher in subjects with GDM as compared to healthy pregnant controls (p < 0.05). Fasting insulin was similar between the two groups. HOMA-IR tended to be higher in GDM group but did not reach statistical significance. Women with GDM had significantly higher triglyceride (p < 0.01) and lower highdensity lipoprotein cholesterol (p < 0.001) than controls. In multiple linear regression analyses, chemerin was significantly associated with BMI (beta-coefficient = 0.274, p = 0.01), HbA1c (beta-coefficient = 0.327, p < 0.01), HDL-cholesterol (beta-coefficient = -0.307, p < 0.01), triglyceride (betacoefficient = 0.236, p < 0.05), insulin levels (beta-coefficient = 0.236, p < 0.05) and HOMA index (beta-coefficient = 0.283, p = 0.01). Conclusions. Maternal chemerin levels were significantly increased in GDM at 24-28 weeks of pregnancy. The physiological significance of elevated serum chemerin in GDM remains unclear.

Objective. This study aims to determine the frequency and prognostic significance of lactic acidosis in children with diabetic ketoacidosis (DKA) admitted to the pediatric intensive care unit. Methods. The study was carried out retrospectively by examining the patients admitted to the pediatric intensive care unit for the treatment of DKA. The ages of the patients ranged from 2 to 18 years. The patients with the following parameters were enrolled in the study: serum blood glucose>200 mg/dL, ketonuria presence, venous blood gas pH ≤7.1, bicarbonate <15. Results. A total of 56 patients were included in the study with a mean age of 111.07 ± 51.13 months. The recovery time from DKA was 16.05 ± 6.25 h in the group with low lactate level and it was 13.57 ± 8.34 h in the group with high lactate level with no statistically significant difference. There was a negative correlation between lactate levels and the recovery time from DKA. Conclusion. Lactic acidosis is common in DKA, and unlike other conditions, such as sepsis, it is not always a finding of poor prognosis that predicts the severity of the disease or mortality. We think that high lactate may even protect against possible brain edema-cerebral damage in DKA.

Context. In patients with suspected primary aldosteronism (PA), the aldosteroneto- renin ratio (ARR) is the most frequently recommended screening test. Further evaluation is based on hormonal changes during volume expansion. Both analyses are critically dependent on an accurate estimation\r\nof renin concentration. Direct active renin (DAR) is a novel laboratory technique used for plasma renin assessment.\r\nObjective. The objective of this study was to evaluate DAR for use in PA diagnostic work-ups.\r\nSubjects and Methods. The study enrolled 69 healthy volunteers. Blood sampling was conducted before and after an\r\nincrease in oral salt intake. Furthermore, a subset of 32 individuals underwent a saline infusion suppression test. DAR and serum aldosterone were measured in all blood samples. To calculate the ARR, serum aldosterone and DAR were expressed in ng/L.\r\nResults. ARR values [median (range); 97.5 percentile] associated with normal and elevated oral salt intake were 8.4 (0.6-37.7); 26.3, and 6.8 (1.1-37.7); 19.6, respectively. DAR and serum aldosterone concentrations\r\n[median (range); 97.5 percentile] after saline infusion suppression were 2.9 (2.7-10.7); 7.2 ng/L and 30 (30-72); 54 pmol/L, respectively.\r\nConclusions. The observed values may be useful in excluding a diagnosis of PA.

Context. Insulin resistance has been detected in a majority of patients with polycystic ovary syndrome (PCOS). Elevated neprilysin levels are associated with insulin resistance. Objective. The present study aims to investigate plasma neprilysin and its relationship with endocrine and metabolic characteristics in patients with PCOS. Subjects and Methods. Thirty-five premenopausal PCOS patients and 35 healthy volunteers of similar age were included in the study. Demographic characteristics, biochemical and hormonal findings and also plasma neprilysin levels were determined in these patients and healthy controls. Results. In our study, HOMA-IR values were significantly higher in PCOS patients (3.3 ± 1.8) compared with the controls [(1.6 ± 1), p<0.01]. Plasma neprilysin levels were significantly higher in the PCOS group compared to the control group (1502.1 ± 1641.2 vs. 764.6 ± 562.6 pg/ mL). There was no difference in plasma neprilysin levels when PCOS patients were classified as overweight-obesity (BMI≥25kg/m2) or non-obesity (BMI<25kg/m2). Conclusion. Our findings revealed significantly higher levels for plasma neprilysin and HOMA-IR values in PCOS patients when compared to controls. No significant differences were noted between obese PCOS patients and non-obese PCOS patients in terms of plasma neprilysin levels.

Context. Lipoprotein-associated phospholipase A2 (Lp-PLA2) is a novel biomarker for cardiovascular diseases (CVD) risk estimation with high specificity for vascular inflammation. Few studies have investigated Lp-PLA2 levels in patients with metabolic syndrome (MetS) and obstructive sleep apnea syndrome (OSAS). Objective. This study aimed to evaluate the role of Lp-PLA2 levels as a marker of vascular inflammation that contributes to cardiometabolic dysfunction in patients with MetS and OSAS. Design. This is a prospective case-control study. Subjects and Methods. 83 men were enrolled. Following anthropometric measurements, laboratory analysis and overnight sleep study, patients were divided into three groups: MetS, OSAS with/without MetS. Serum Lp-PLA2 levels were determined by ELISA method. Results. Serum Lp-PLA2 levels were statistically significant among the three groups and were higher in OSAS with MetS group than those without MetS. A significant positive relationship between increased Lp-PLA2 level and CRP (C-reactive protein) and apnea–hypopnea index (AHI) was found. Average oxygen saturation (AvO2) and the lowest oxygen saturation were negatively correlated with Lp-PLA2. The number of desaturation events, oxygen desaturation index, AvO2, AHI and CRP were significant predictors of Lp-PLA2. Conclusions. Lp-PLA2 levels are associated with OSAS severity and might play an important role in predicting CVD in OSAS with/without MetS

Context. Papillary thyroid carcinoma (PTC) is the most common type of thyroid cancer due to its high fibrotic content; it can affect the blood flow resistance. Objective. To introduce duplex Doppler ultrasonography (DDUS) parameters of papillary thyroid carcinoma (PTC) and their correlation with size. Design. The study was designed as a prospective study. Subjects and Methods. Thyroid nodules of the patients who were already scheduled for thyroid surgery either for malignant thyroid nodules or multinodular goiter were evaluated for DDUS parameters. Size, systolic to diastolic flow velocity (S/D) ratio, pulsatility index (PI), and resistive index (RI) of each nodule were recorded. Nodules were diagnosed as PTC or benign nodules based on histopathology. DDUS parameters were compared between PTCs versus benign nodules and micro PTCs (≤ 10 mm) versus large PTCs (> 10 mm). A correlation analysis was performed between the size and DDUS parameters. Results. 140 thyroid nodules (30 PTCs, 110 benign nodules) were obtained. The mean S/D ratio, PI, and RI values were significantly higher in PTC than in benign nodules (p values were 0.0001, 0.0003, and 0.0001 respectively). The optimal cut-off values of S/D (0.732), PI (0.732), and RI (0.738) had accuracy rates of 71%, 69%, and 69%, respectively. There was no statistically significant difference between micro PTC and large PTC with regards to DDUS parameters. The size and DDUS parameters of PTC showed no significant correlation. Conclusions. PTC has a high resistive flow pattern regardless of its size; however the clinical utility of DDUS to differentiate a PTC from benign nodule is limited.