ACTA ENDOCRINOLOGICA (BUC)

Objective. To observe the impact of quercetin and isoquercitrin on gluconeogenesis in hepatocytes. Methods. Mouse primary hepatocytes were cultured with lactic acid and pyruvic acid. After treatment with quercetin and isoquercitrin for 24 hours, the glucose concentration in the culture supernatant was determined. RT-PCR was used to detect the mRNAs of PEPCK, G6Pase, LKB1, and AMPKα. Protein levels of LKB1, AMPKα, and Thr172 phosphorylation were evaluated by Western blot. Results. The glucose concentration in the gluconeogenesis group (GN) was significantly higher than in the control group (C), but the glucose concentrations in the high level quercetin(group 80Q) and high level isoquercitrin (group 80I) were significantly lower than in the group GN, P<0.01. In the group 80Q, and group 80I, the mRNA levels of PEPCK and LKB1were significantly lower than in the group GN (P<0.01), and the G6Pase mRNA were significantly lower than in the group GN (P<0.05). The protein levels of LKB1 and the phosphorylation of AMPKα Thr172 in the group 80Q, group 40I, and group 80I were higher than in the group GN. The effects of quercetin and isoquercitrin on LKB1 and AMPKα were similar to those of metformin. Conclusions. Quercetin and isoquercitrin inhibit gluconeogenesis in hepatocytes, which may be related to the LKB1 upregulation and phosphorylation of AMPKα.

Introduction. Immune Dysregulation, Polyendocrinopathy, Enteropathy X-Linked (IPEX) Syndrome represents a rare X linked disorder, characterised by development of systemic autoimmunity from the first year of life. IPEX is due to mutations in the FOXP3 gene located on the X-chromosome. There are no specific laboratory tests to confirm the diagnosis. Molecular analysis of the FOXP3 gene(Xp11.2-q13.3) is required for the diagnosis. Chronic immunosuppression and hematopoietic stem cell transplantation represent the two main therapeutic interventions for this immune dysfunction, the last one being the only curative treatment. Case presentation. The authors present the case of a male patient aged 14 years old, who was admitted for ketoacidotic diabetes onset. He was diagnosed with polyendocrine autoimmune association (diabetes mellitus type 1, autoimmune thyroiditis and hypo-gonadotrophic hypogonadism). This patient associated celiac disease, sustained on clinical, immunological and histological changes:recurrent diarrhea, positive IgA anti-tissue transglutaminase antibodies and total villous atrophy on intestinal biopsy sample. He also presented recurrent eczematous dermatitis associated to elevated serum concentration of immunoglobulin E. The authors sustained the diagnosis of IPEX syndrome in this case based on family history of unexplained early deaths of the patient´s uncles from the motherside, along with clinical and laboratory aspects. Drug treatment included nutritional support, immunosuppressant therapy and hormone replacement. Conclusions. The most important aspect in this case was considering IPEX syndrome after integration of all clinical and paraclinical data but without molecular analysis of the FOX P3 gene. The presumption of IPEX syndrome reconsidered in this case the treatment and the prognosis. Life expectancy is reduced in this condition that usually occurs in the first months of life. The particularity of this case was the late onset of IPEX syndrome, presenting a severe phenotype with aggressive autoimmune associations that led finally to the patient’s death.

Prolonged target gland failure causes pituitary hyperplasia, but rarely, secondary hyperplasias develop into autonomous neoplasms. We report herein a rare example of gonadotroph adenoma arising in a patient with prolonged hypogonadism due to Klinefelter syndrome. A pituitary macroadenoma with suprasellar extension was discovered incidentally by magnetic resonance imaging (MRI), in search for the cause of chronic saliva retention. His pre-operative serum concentrations of both luteinizing hormone (LH) and mostly follicle-stimulating hormone (FSH) were distinctly higher than normal, as expected, but the levels decreased after complete removal of the tumor, suggesting partial secretion of gonadotropins by the tumor. The surgically removed tissue showed a typical pituitary adenoma with distinct immunoreactivity for FSH (intense, homogeneous) and LH (scattered). In the fragmented parts of adjacent gland tissue, no hormone producing cell hyperplasia or presence of gonadal deficiency cells were detectable. In conclusion, our case is the description of a rare example of gonadotropin producing pituitary adenoma (FSH and LH) with increased serum levels of both gonadotropins in a patient with untreated Klinefelter syndrome.

Context and objective. In this study, we aimed to investigate how moderate physical activity improves the bone ultrastructural parameters in rats with glucocorticoidinduced secondary osteoporosis. Animals and Methods. Research has been carried out on Wistar female rats. Secondary osteoporosis was induced through daily i.m.1.5 mg/kgbw methylprednisolone, over a period of 30 days. A group of rats with induced secondary osteoporosis were subjected to physical activity (swimming) for one hour/day for 30 days. Rats were sacrificed 24 hours after the last administration and femoral bones were used for electron microscopy analysis. Results. The ultrastructural findings obtained from the rats with osteoporosis showed varying degrees of alteration in all cellular components. A moderate physical effort led to the overall maintenance of the normal ultrastructure of the cells and connective components, protecting the lamellar structure of the compact bone from the deleterious effects of glucocorticoid. The shape and components of osteocytes were also preserved and the accumulation of lipids in the bone marrow diminished. Conclusions. Physical exercise has been shown to have a protective role by lowering the development of structural alterations specific to osteoporosis. Therefore, moderate physical exercises are recommended for improving the structure of the bone mass affected by glucocorticoid treatment.

Context. We propose that the underlying etiology of renal calcium leak is complex and involves defects in renal handling and parathyroid sensing of ambient calcium concentration in the tubular fluid and blood. Therefore, treatment of such a patient requires both decreasing the parathyroid mass and inhibiting calcium sensing receptors that are present in the parathyroid and kidney. However, a combined treatment strategy of three-gland parathyroidectomy and calcimimetic therapy has not been formally studied to date. Objective. To present a patient with renal calcium leak causing secondary hyperparathyroidism presenting as primary hyperparathyroidism. There are a two year followup period. Results. A patient with mild hypercalcemia, hypercalciuria, musculoskeletal pain, and recurrent kidney stones underwent a three gland parathyroidectomy and had persistent hypercalciuria post-operatively. She was subsequently treated with thiazide diuretic that caused dramatic decrease in hypercalciuria, but overt hypercalcemia. She was then treated with Cinacalcet with normalization of intact PTH, serum calcium and serum phosphate. Conclusion. Patients with hypercalciuria and mild hypercalcemia may have secondary hyperparathyroidism. Renal calcium leak drives hyperparathyroidism and is unresponsive to parathyroidectomy or thiazide diuretic alone. In our patient, three gland parathyroidectomy plus calcium –sensing mimetic agent, Cinacalcet, normalized serum calcium, PTH, and phosphorus. Defects in calcium sensing in the parathyroid gland and kidney might be responsible for this form of secondary hyperparathyroidism.

Objective. The aim of the present study was to evaluate the effect of electromagnetic field (EMF) exposure during developmental period on parameters of oxidative stress and histopathology of testis and testosterone level in adult rat F1 generation. Methods and study design. In treatment group pregnant rats were exposed to 3mT EMF, 50Hz for 21 days. The sham group contained pregnant rats under same condition, but out off the EM field. Pregnant rats in room were used as control group. After delivery, the blood samples of mothers for biochemical analyses of total antioxydant capacity (TAC) and malondialdehide (MDA) were provided. The male pups were kept until maturity, then their TAC, MDA and testosterone levels were analyzed, also their tests were removed for investigation of histopathology changes with light microscopy. Results. Biochemical analysis showed that TAC and MDA was significantly increased in pregnant rats in the treatment group when compared with the control group (p<0.05). In adult male of F1 generation MDA was significantly increased in treatment group, but TAC and the testosterone level was significantly decreased in the treatment group as compared with the control group (p<0.05). Microscopic results revealed that in experimental group seminiferous epithelium contained many small irregular empty spaces as the sign of cellular sloughing, spermatogenic cells appeared to be disrupted. The nuclei of spermatogonia cells were heterochromatic, also dense of germinal epithelium and the number of spermatozoa was decreased. Conclusion. The results of this study suggest that pregnant rats exposure in EMF led to oxidative stress in adult male of F1 generation and showed adverse effect on testosterone and spermatogenesis in adulthood which may produce subfertility.