Corin Badiu

Localization and diurnal expression of mRNA encoding the β1-adrenoceptor in the rat pineal gland: an in situ hybridization study

Abstract.The rat pinealocyte is stimulated by norepinephrine, which is released from sympathetic nerve fibers innervating the gland. Norepinephrine binds to β1-adrenoceptors situated on the pinealocyte cell membrane. Ligand binding to these receptors exhibits a diurnal rhythm, with the largest number occurring in the late part of the light phase when the release of norepinephrine is minimal. By using in situ hybridization with a cDNA antisense oligonucleotide probe recognizing mRNA encoding the rat β1-adrenoceptor, we have demonstrated a stronger hybridization signal in the rat pineal gland than in other brain regions. Cells containing β1-mRNA are located in the superficial pineal gland, the deep pineal gland, and the pineal stalk. However, the number of receptors varies considerably between the individual pinealocytes. The β1-mRNA in situ hybridization signal for mRNA encoding the β1-adrenoceptor of the rat pineal has been quantified over a 24-h period; the strongest signal is found at mid-dark and the weakest signal at mid-light, indicating that the transcriptional regulation of β1-mRNA synthesis in the rat pineal is diurnal. In addition, maximal receptor protein expression occurs late in the light phase in the rat pineal and is thus considerably delayed compared with its peak mRNA synthesis. This lag time indicates that the β1-receptor is regulated at the translational or post-translational level. Removal of the sympathetic input to the pineal gland by superior cervical ganglionectomy decreases the β1-mRNA signal in the gland.

Presence of oxytocinergic neuronal‐like cells in the bovine pineal gland: an immunocytochemical and in situ hybridization study

In the last decade, there is more and more evidence showing the role of the central innervation of the pineal gland, but there are controversies around the intra or extrapineal origin of oxytocin found within the pineal tissue. In order to check the amount and the site of synthesis of oxytocin in the bovine pineal gland, we performed a morphological and chromatographic study. The anatomical distribution of the pineal oxytocin was explored by immunohistochemistry and in situ hybridization for the corresponding mRNA. The results confirm the presence of oxytocinergic fibres in the bovine pineal, some of them endowed with big varicosities. Immunohistochemistry also displayed neuronal‐like cells in the pineal body. The in situ hybridization for the mRNA encoding pre‐pro/oxytocin‐NFZ I used a mixture of three oligonucleotide probes labelled with 35S. This allowed identification of positive cells in the bovine pineal. The content in oxytocin was evaluated by radioimmunoassay during 5 months, from July to November, and the peptidic extract revealed an increase of pineal oxytocin immunoreactivity in September as compared with July or November. The significance of intrinsic oxytocin innervation of the bovine pineal gland, as well as the threefold increase of the oxytocin content in the pineal in September, remains to be elucidated.

Pharmacological management of non-alcoholic fatty liver disease: Atorvastatin versus pentoxifylline

In this study, we aimed to evaluate the efficacy of pentoxifylline and atorvastatin in the treatment of non-alcoholic fatty liver disease (NAFLD). The study included 98 patients with histologically confirmed NAFLD divided into 2 groups as follows: group I (57 dyslipidemic patients, receiving atorvastatin 20 mg/day and group II (41 non-dyslipidemic patients, treated with pentoxifylline, 800 mg/day). The present study was conducted for a mean of 32.8±3.4 weeks. For all patients, we determined the body mass index, a liver biopsy was performed, and we measured the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT), alkaline phosphatase (ALP), total cholesterol (TC) and triglycerides (TG) at the beginning and at the end of the study period. The NAFLD activity score (NAS) was used to evaluate the liver biopsies for steatosis, fibrosis and necroinflammation. The patients in group I exhibited a considerable reduction in ALT, AST, GGT, TC, AP and TG levels (P<0.0001). Histologically, there were no changes in fibrosis and necroinflammation, although the extent steatosis was reduced. The improvement in the ALT, AST and GGT values (P<0.05) in group II were similar to those in group I; however, no statistically significant decrease was noted in the levels of ALP, TC and TG in this group. Our results thus demonstrated that atorvastatin attenuated steatosis and improved liver function parameters in patients with NAFLD associated with dyslipidemia. Similar results were obtained in the non-dyslipidemic patients administered pentoxifylline.

Age-dependent endocrine disorders involved in the pathogenesis of refractory acne in women

Acne is a disorder of the pilosebaceous unit, common among adolescents, which may be extended to adulthood. The aim of this study was to assess the prevalence of hormonal disorders in women with acne resistance to conventional therapy. We included 72 women aged between 15 and 36 years (divided in two age groups) who presented to our clinic between May and October 2014, suffering from moderate and severe forms of papulopustular and nodulocystic acne. The subjects were non-responsive to classic dermatological treatment or had clinical manifestation of hyperandrogenism. Based on age, we divided the women into two groups, group I with 40 patients aged 15–22years and group II with 32 patients aged 23–36 years. Using ELISA, a hormonal profile was performed for each patient in days 1–3 of the menstrual cycle including, total testosterone, dehydroepiandrosterone sulfate (DHEA-S), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol, prolactin, and plasma cortisol. For statistical analysis we used Stata 13 software. We compared the hormonal profile of the two groups and identified significant differences for: testosterone levels (mean value, 0.64±0.35 vs. 0.97±0.50 ng/ml; p<0.0001), DHEA-S levels (mean value, 0.85±0.27 vs. 1.05±0.33 mg/24 h; p=0.001), prolactin levels (mean value, 281.85±91.113 vs. 353.969±102.841 mIU/ml; p=0.002) and LH levels (14.8±6.7 vs. 20.1±8.2 mIU/ml; p=0.002) were higher in group II. No statistically significant differences were found for estradiol (p=0.588) and cortisol (p=0.182) levels. In conclusion, refractory acne can be the first sign of systemic illness including polycystic ovary syndrome. Thus, for a correct therapeutic approach it is necessary to interpret the clinical and biochemical elements in correlation with the medical history.

Mitochondrial DNA haplogroup K as a contributor to protection against thyroid cancer in a population from southeast Europe

We aimed to analyze the contribution of mitochondrial DNA (mtDNA) haplogroups of the mtDNA control region to thyroid cancer risk in a population from southeastern Europe consisting of 235 thyroid tumor patients, including 114 patients with thyroid follicular adenoma, 121 patients with papillary thyroid carcinoma, and 419 healthy controls. Binary logistic regression with adjustment for age and gender revealed that mtDNA haplogroup K was significantly associated with a protective role for thyroid cancer in the combined tumor group versus controls. These results indicate a potential role for mtDNA haplogroups as important candidate susceptibility markers for the patients with thyroid nodules.

Branched-Chain Amino Acid Database Integrated in MEDIPAD Software as a Tool for Nutritional Investigation of Mediterranean Populations

Branched-chained amino acids (BCAA) are essential dietary components for humans and can act as potential biomarkers for diabetes development. To efficiently estimate dietary intake, we developed a BCAA database for 1331 food items found in the French Centre d’Information sur la Qualité des Aliments (CIQUAL) food table by compiling BCAA content from international tables, published measurements, or by food similarity as well as by calculating 267 items from Greek, Turkish, Romanian, and Moroccan mixed dishes. The database embedded in MEDIPAD software capable of registering 24 h of dietary recalls (24HDR) with clinical and genetic data was evaluated based on archived 24HDR of the Saint Pierre Institute (France) from 2957 subjects, which indicated a BCAA content up to 4.2 g/100 g of food and differences among normal weight and obese subjects across BCAA quartiles. We also evaluated the database of 119 interviews of Romanians, Turkish and Albanians in Greece (27–65 years) during the MEDIGENE program, which indicated mean BCAA intake of 13.84 and 12.91 g/day in males and females, respectively, comparable to other studies. The MEDIPAD is user-friendly, multilingual, and secure software and with the BCAA database is suitable for conducting nutritional assessment in the Mediterranean area with particular facilities for food administration.

Common Features of Giant Prolactinoma and Paranasal Neuroendocrine Carcinoma-Case Report

Giant prolactinoma is a very rare tumor that due to its massive extension into surrounding structures can present more often with neurological complications such as visual defects, cranial nerve paresis or even hydrocephalus, unlike the classic prolactinoma presentation with amenorrhea, infertility and galactorrhea. On CT/MRI exams it can present as aggressive skull base tumor and its immunohistochemistry (IHC) may have common features with neuroendocrine neoplasms.

Heterogeneous presentation of giant prolactinoma

Background: Giant prolactinomas are relatively rare pituitary tumors, defined as adenomas greater than 4 cm, with extrasellar extension, presenting with PRL levels > 1000 ng/ml. Dopamine agonists (DA) are the first-line treatment. Most (90 %) of giant prolactinomas occur in men, presenting with a symmetric distribution with a peak during the forth decade of life. It appears that men are younger at diagnosis and with larger tumor size, not only due to a longer delay before diagnosis, but rather to an overall greater growth potential of prolactinoma in males.

Current status and perspectives in the treatment of Prader-Willi syndrome

Introduction: Prader-Willi syndrome (PWS) is a neurometabolic genetic disorder with chromosomal 15q11-q13 deletions or uniparental disomy with absence of paternally expressed genes affecting 1/12,000 – 1/15,000 newborns. The syndrome is characterized by hypotonia, intellectual disability, behavioral problems, psychiatric phenotypes, short stature, central hypocorticism, pubertal insufficiency and hypogonadism leading to osteoporosis. The most life-threatening characteristic is excessive appetite, which leads to morbid obesity and diabetes and its related complications. Areas covered: In this paper, the authors review evidence-based papers that present the management of patients with PWS, involving aspects of reduced GH secretion, hypogonadotropic hypogonadism, abnormal appetite control and high pain threshold, all suggesting hypothalamic-pituitary dysfunction. The treatment of these homeostatic alterations is presented, and the consideration of a multidisciplinary approach and future perspectives are discussed. Finally, the authors briefly review the current status of treatment and perspectives specific for PWS. Expert opinion: Despite the detailed knowledge about obesity mechanisms regulated at the hypothalamic level, the pharmacological intervention is limited currently to substitution of proven endocrine deficiencies (growth hormone, thyroxine, sex steroids, glucocorticoids), prevention and treatment of metabolic complications (the most severe being diabetes) and osteoporosis. Treatment of mental disturbances and skin-picking, severe obesity and altered body composition often requires a team including a pediatrician, an endocrinologist, a psychiatrist, a bariatric surgeon and strong support from the family and caregivers.

Quinagolide Effects in Normoprolactinemic Galactorrhea

To the Editor: Galactorrhea, the normal breast secretion, is encountered mostly in women with hyperprolactinemia. Physiological galactorrhea occurs in postpartum circumstances; besides, very high prolactin levels are encountered in secreting pituitary adenomas (prolactinomas and acromegaly) or in so-called functional hyperprolactinemia with a myriad of pharmaceutical or noniatrogenic causes. However, in rare cases, galacthorrea occurs in euprolactinemic women. Prolactin (PRL) is a single-chain polypeptide of 198 amino acids with a molecular weight of 23 kilodaltons. Big prolactin is the result of glycosilation (25–27 kDa) that can dimerize or trimerize with sizes of 50–60 kDa, while big-big prolactin/bb-PRL (macroprolactin) can be in part a complex of 23 kDa PRL with an IgG (1). According to current concept, macroprolactin (less than 20% of PRL immunoreactivity in normal circumstances) is biologically inactive and, therefore, its accumulation in serum has little, if any, pathological significance (2). Apart from its classical functions, stimulation of lactogenesis and galactopoesis, PRL affects other aspects of body function, including osmoregulation, metabolism, and regulation of the immune and the central nervous system (3). Prolactin is secreted continuously at low basal rates throughout life, regardless of sex and its concentration in blood increases during nocturnal sleep, or in an episodic manner in response to eating and stress. Basal values are somewhat higher in women than in men, presumably reflecting the effects of estrogens. Considering the significant heterogeneity of PRL’s molecule, laboratory assays can give different results due to glycosylation, degradation or other actions; the nonglycosylated form of prolactin is usually dominant. There is gross variability in the detection of prolactin in sera-containing big-big prolactin by commercial immunoassays (4). Macroprolactinemia is characterized, by most authors, as a benign condition with no clinical implications (5), although a number of investigators challenged this view, suggesting that in some cases, with mild symptomatology, a therapeutic intervention is possibly required (6). Therefore, in hyperprolactinemic patients (idiopathic or from a prolactinsecreting pituitary microadenoma), pharmaceutical treatment is often required. This consists of a variety of dopamine agonists, including cabergoline, bromocriptine, and nonergotic quinagolide. Nowadays, quinagolide’s therapeutic indications include idiopathic hyperprolactinemia or hyperprolactinemia from a prolactin-secreting pituitary microadenoma or macroadenoma. In a multicentric study, we aimed to assess the efficiency of quinagolide in patients with galactorrhea having normal (or borderline) prolactin levels. The patients were informed about the study and gave their informed consent. The institutional ethics committees approved the study protocol. Thirty-four patients in four clinics in Romania and Greece with bilateral galactorrhea were included initially in the study. Twenty-four patients (29–51 years, mean age 38.1) met the inclusion criteria (prolactin levels <30 ng/mL, bilateral galactorrhea, nonpregnant, no lactation, no hypothyroidism: T3, T4, TSH normal levels, no medicine taken). Prolactin was measured using the commercially available assay from NovaTec Immundiagnostica GmbH (Dietzenbach, Germany) and ranged from 10.3 to 29.4 ng/mL (mean 19.9 ng/mL). Galactorrhea was graded clinically as + (only drops from the nipple after pression [five patients]), ++ (spontaneous galactorrhea [10 patients]) and +++ (continuous galactorrhea with spots in the bra [nine patients]). Quinagolide was given for 3 months at gradually increased doses within 2–3 weeks, achieving a final dose of 75 lg/day. Breast ultrasound was performed before and after Quinagolide with measurement of diameter of largest ducts. Final Address correspondence and reprint requests to: Corin Badiu, Institute of Endocrinology, Blvd. Aviatorilor 34-36, Bucharest 011863, Romania, or e-mail: badicrin@yahoo.co.uk