ACTA ENDOCRINOLOGICA (BUC)

Background. We aimed to evaluate whether a high carbohydrate or a high fat diet differs in alteration of the inflammatory and metabolic risk factors in cardio-renal metabolic syndrome in rats. Methods. Twelve male Wister rats were randomly divided into two groups: one received diet 1 standard pellet rat diet (D1) containing 10% fat, 50% carbohydrate, 25% protein and another group received diet 2 (D2) containing 59% fat, 30% carbohydrate and 11% protein for 16 weeks. Weight was recorded weekly. FSG and insulin levels were measured using an enzymatic spectrophotometric and a standard ELISA kit respectively. Inflammatory parameters including TGF-β, MCP-1, TNF-α, IL-1β, IL-6 in the renal and cardiac tissues of rats were evaluated by ELISA technique. Result. Food intake in D1 and D2 groups increased in the study period, however food intake in D2 group was significantly higher compared with D1 group. FSG, HOMA and TG concentrations in D2 group were significantly higher compared to D1 group. Moreover, TGF-β and MCP- 1 concentrations in the renal tissues of D2 group and TNF-α in the cardiac tissues of D1 group were significantly higher compared with D1 group (P<0.05). Positive associations between IL-1β and TG and between HOMA, FSG with TGF-β and MCP-1 in the renal tissue of animals were also identified.

Background. Saturated free fatty acids, such as palmitate, can cause pancreatic β-cell apoptosis. Although the toxicity of palmitate could be mediated partly through endoplasmic reticulum (ER) stress, the mechanism by which fatty acid over-accumulation led to lipoapoptosis in β-cells has not been fully understood. Recently, the fat mass and obesity associated (FTO) gene is proved to be related to obesity and type 2 diabetes, but its function in β-cells remains largely unknown. Whether or not FTO could counteract palmitate induced β-cell apoptosis remains to be investigated. Methods. INS-1 cells were infected with FTO expression adenovirus and incubated with palmitate. Then, viability and induction of apoptosis were measured by MTT assay and Hoechst-staining, respectively. Western blot analyses were performed for unfolded protein response specific proteins and mRNA expression of target molecules was determined by real time-PCR. Results. Palmitate incubation led to β-cell apoptosis, whereas adenovirus-mediated FTO overexpression significantly ameliorated the effect of palmitate. Increased activation of X-box binding protein 1 (Xbp1) mRNA and phosphorylation of eIF2α were also observed after palmitate treatment, whereas FTO overexpression significantly ameliorated the effect of palmitate. The proapoptotic transcription factor CHOP was significantly enhanced by palmitate incubation. In contrast, in accordance with sustained cell survival, FTO overexpression resulted in notably decreased CHOP levels. Interestingly, mRNA expression of the chaperones Pdi, Calnexin and Grp94 was not altered by palmitate treatment, while FTO overexpression notably increased the expression of Bip. Conclusion. Our data showed that FTO overexpression could protect β-cells from palmitate-induced apoptosis partly through suppression of ER stress.

Context. Some adrenal tumors, such as pheochromocytoma, can be life-threatening. Therefore it is crucial to distinguish them from other lesions, especially prior to surgery. Chromogranin A (CgA) seems to potentially be a good marker for tumors of chromaffin origin. Objective. To assess the differentiating value of CgA in the diagnostic work-up of pheochromocytoma. Design. Retrospective study of operated patients with adrenal incidentaloma with lesions > 10 Hounsfield’s units (HU) on CT. Subjects and Methods. Thirty patients (11 males, 19 females; aged 61.5±21 years) were enrolled in the study. Patients using medications interfering with the assessment of CgA and metanephrines were excluded. Two groups were formed: those with pheochromocytoma (Ph, n=16) and those with non-pheochromocytoma (N-Ph, n=14) lesions. Data included radiological features of masses, serum CgA and 24-hour urine metanephrines (24 - HUM) concentrations. Results. No difference in 24-HUM level nor tumor size or density was found between groups Ph and N-Ph. Median serum CgA concentration was higher in Ph group compared to the N-Ph: 99.35 (68.12-172.73) vs. 52.92 (34.37-101.26) ng/mL, respectively (P=0.04). In Ph group, the size of the lesion correlated negatively with density (r= -0.53, P=0.042). No significant correlation in CgA, 24-HUM, density or size of the lesion was found. Performed curve receiver operating characteristic (ROC) showed AUC=0.7232 for CgA. Taking into account CgA serum value of ≤ 50 ng/mL (sensitivity: 93.75%, specificity: 50.00%, P=0.012), we proposed an algorithm for management of lesions > 10 HU on CT. Conclusion. CgA level ≤ 50 ng/mL might be useful in initial screening evidence for the exclusion of pheochromocytoma. It is crucial to eliminate factors interfering with the measurements.

Nodular thyroid disease (NTD) is represented by palpable thyroid nodules (solitary, multiple) and thyroid incidentalomas (identified by means of thyroid ultrasonography). The discussed entities carry the same risk of malignancy (about 5 %). The main objective in evaluating NTD is represented by the exclusion of malignancy by means of corroborated investigations, focused on the value of a panel of IHC markers.\r\nMaterial and methods. We included in the study 27 cases of NTD, evaluated by means of: clinical investigation, ultrasonography of the thyroid, cytological examination, morphological analysis and IHC. The used panel of IHC markers comprised: Ki-67, PCNA, CK 19 and c-erbB2 (DAKO LSAB method)\r\nResults. From the total of cases, 8 presented positivity with Ki-67 and 17 with PCNA. Regarding CK 19, the majority of PTC cases stained ++ and diffusely, but not papillary hyperplasia (focal positivity).c-erbB2 diffuse and intense positivity (+++) was noticed in PTC.The case with a follicular tumor of uncertain malignant potential stained weakly only with c-erbB2.\r\nConclusions. From the used panel of IHC markers, CK 19 presented the best value, being able to differentiate FVPTC from FTC and PTC from papillary hyperplasia.

Various clinical and experimental studies indicate that gonadal hormones exert\r\nmodulatory effects on nociception and analgesia.\r\nThe aim of the present study was to investigate the role of gonadal hormones in the\r\nresponse by male and female rats to thermal nociceptive stimulation.\r\nMaterials and Methods. 7 week old albino (Wistar) rats were used in our study.\r\nAnimals were randomly divided into control, sham and ovariectomised or orchidectomised\r\ngroups. Thermal pain threshold was measured through tail immersion test before and 10, 20\r\nor 40 days after gonadectomy. The pain threshold was measured as the time required to elicit\r\na flick of the tail called analgesia time. Serum testosterone, estradiol, progesterone or\r\nprolactin levels were measured simultaneously.\r\nThe results showed that analgesia time was higher in female (5.11 min) than in male\r\n(4.93 min) intact animals (p<0.05) indicating sex difference in response to thermal\r\nnociception. Serum testosterone, estradiol or progesterone level as well as analgesia time\r\nwere not significantly reduced 10 days after gondectomy. In male animals, analgesia time\r\nwas significantly decreased (p<0.01) 20 or 40 days after orchidectomy (2.25 or 2.14 min,\r\nrespectively) compared with control rats (4.93 min). Serum testosterone concentration was\r\nsignificantly reduced (p<0.05) 20 or 40 days after orchidectomy (0.08 or 0.09 ng/mL,\r\nrespectively) compared with control serum testosterone level (2.14 ng/mL). In female rats,\r\nanalgesia time was significantly decreased (p<0.001) 20 or 40 days after ovariectomy (2.87\r\nor 2.66 min, respectively) compared with control rats (5.11 min). Serum estradiol\r\nconcentration was significantly reduced (p<0.001) 20 or 40 days after ovariectomy (3.17 or\r\n0.87 ng/mL, respectively) compared with control serum estradiol level (19.95 ng/ml). Serum\r\nprogesterone level was also decreased (p<0.001) 20 or 40 days after orchidectomy (5.27 or\r\n0.55 ng/mL, respectively) compared with control serum estradiol level (17.66 ng/mL).\r\nSerum prolactin level was not significantly enhanced during experiment indicating that there\r\nwas not heat stress influencing the procedure.\r\nConclusively, our findings clearly indicate that depletion of gonadal hormones 20 or 40 days\r\nafter gonadectomy modulates the pain-induced behavioral responses related to thermal nociception.

Background. If not diagnosed at birth, congenital hypothyroidism (CH) can cause deleterious, irreversible neurodevelopmental sequels. The importance of thyroid newborn screening (NBS) is therefore well established. Objective. To evaluate the efficacy of NBS for CH in North-East Romania. Methods. Retrospective, descriptive study involving 271662 newborns screened between 2010 and 2019 for CH and phenylketonuria in maternities from six Romanian NorthEastern counties by measuring neonatal TSH (neoTSH) in the whole blood extracted from the heel between days 3 and 5 after birth. Values found higher than a cut-off level of 10 mIU/L were followed by serum evaluation of TSH and fT4 for the confirmation of CH. Thyroid ultrasound was further performed at children found with CH. Results. NeoTSH was found elevated in 417 newborns, but CH was subsequently confirmed in only 57 cases (1/4766 newborns). Mean age at the time when diagnosis was communicated was of 37.2 ± 15 days (between 9 and 157 days). Mean age when therapy was started was of 44.2 ± 17.9 days (between 13 and 160 days) with a mean delay of one week from diagnosis (between 0 and 62 days). Thyroid ultrasound revealed athyreosis in only 3 cases, atrophic thyroid gland in other 10 cases, whereas the thyroid was described as present in the remnant 44 cases. The number of first year follow-up visits greatly varied from 0 to 5, with an average of 2. Conclusions. NBS allowed rapid diagnosis of CH in North East Romania. The communication of diagnosis to families and therapy onset were however often delayed. Diagnosis and therapy onset before the age of two weeks, as well as a tighter follow-up should be assured by the healthcare system. Etiological diagnosis should be more accurate, for a better prognosis of disease severity, as well as the possibility of genetic advice in selected cases.

Context. It is unclear whether treatment is necessary for transient moderate hypocalcaemia occurring after total thyroidectomy; if it is present, it is unclear which treatment modality should be preferred. Objective. To investigate both the necessity and effectiveness of different treatment approaches of oral and/ or intravenous calcium treatment in patients with transient, postoperative, moderate hypocalcaemia. Design. This is a case control study made between June 2014 and June 2015. Subjects and Methods. Forty-five patients who had serum calcium levels 6 hours after total thyroidectomy between 7.5-8 mg/dL were divided into three equal groups: an oral calcium administration group, an intravenous calcium administration group and a no-treatment group. Serum calcium and parathyroid hormone levels were measured preoperatively and on postoperative days 1, 2, 5 and 10. Results. For post-thyroidectomy patients with serum calcium 7.5-8 mg/dL in the early postoperative period, no significant difference in serum calcium or parathyroid hormone was detected between groups. Conclusions. Follow-up without treatment seems to be the most effective approach for moderate hypocalcaemia occurring in the early period following total thyroidectomy; this suggests that intravenous treatment should be avoided.

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Objective. In postmenopausal period, changes in bone turnover markers (BTM), vitamin D3, cytokines and parathyroid hormone (PTH) are frequently observed. The study was to assess bone mineral density (BMD) and bone metabolism index (IBM) in the perimenopausal women. Design years: 2013-2014. Subjects and Methods. One hundred and thirteen women were divided into four groups: group I (35 not menstruating 50 - 60 years old with osteoporosis), II (23 not menstruating 50 - 60 years old without osteoporosis), III (30 menstruating 40 - 49 years old with osteoporosis), IV (25 menstruating 40 - 49 years old without osteoporosis). The following parameters were measured: IL-1β, IL-6, TNF-α, hormone oestradiol (E2), PTH, FSH, TSH, calcium (Ca2+), phosphates (P), alkaline phosphatase (bALP), C-terminal telopeptide of type I collagen alpha 1 chain (α1CTX), osteocalcin (OC), BMD, IBM. Results. IBM and BMD were significantly lower in premenopausal than in postmenopausal women. The concentration of OC, CTX, 25OH D3 and PTH levels differed significantly between group I vs. II, group I vs. III and group II vs. IV. Conclusions. The levels of BTM, D3, PTH differed significantly between groups. This study demonstrated that bone metabolism depended mainly on processes related with menopause state and changes in D3, PTH and cytokines levels.