Purpose: Gastroenteropancreatic neuroendocrine tumors (GEP-NET) are classified by the WHO, yet its prognostic value needs to be confirmed. Therefore, we aimed to determine the prognostic role of cell cycle key regulatory genes p53, p27 kip1 (p27), and cyclin E in this tumor entity. Experimental Design: Tumor specimen from 89 patients with a complete follow-up were studied immunohistochemically for p27 and cyclin E expression and for p53 mutations. The functional relevance of p27 was evaluated in the neuroendocrine cell lines BON1 (human) and INS1 (rat) by the use of small interfering RNA. Results: Twenty-six of 29 benign, well-differentiated endocrine tumors (WHO class 1) showed a high expression (>50%) of p27, whereas all 10 poorly differentiated endocrine carcinomas (WHO class 3) displayed a low expression of p27. Metastatic well-differentiated endocrine carcinomas (WHO class 2) showed a low p27 expression in 20 of 50 (40%) patients, which conferred a poor prognosis (median survival, 57 versus 140 months; P = 0.037). This prognostic dichotomy was improved by the use of a combination of p27 and cyclin E (high cyclin E/low p27 versus low cyclin E/high p27: median survival 53 months versus not reached; P = 0.0044). p53 mutations were rare (1of 10 poorly differentiated endocrine carcinomas). Conclusions: Loss of p27 and overexpression of cyclin E play a critical role in the aggressiveness of gastroenteropancreatic neuroendocrine tumors. This coincides with increased cell cycle progression. We propose a discussion whether to incorporate the immunohistochemical expression of p27 into a revised classification to individualize therapeutic strategies in this tumor entity.

Background: Therapeutic approaches to gastroenteropancreatic neuroendocrine tumors (GEP-NETs) are still not satisfactory. A new direction in treatment options could be the novel aurora kinase inhibitor ZM447439, which was previously reported to interfere with the mitotic spindle integrity checkpoint and chromosome segregation, but does not interfere with other kinases when used up to 5 µM. Methods: We evaluated the antineoplastic effects of ZM447439 on growth and apoptosis of the GEP-NET cell lines BON, QGP-1 and MIP-101, representing the different malignant tumor types, using standard cell biological tests as crystal violet assays, caspase activation, DNA fragmentation and cell cycle analysis. Results: ZM447439 dose-dependently inhibited proliferation of all three cell lines with IC₅₀ values in the nanomolar to low micromolar range. Moreover, aurora kinase inhibition by ZM447439 potently induced apoptosis, which was accompanied by DNA fragmentation and caspase 3 and 7 activation. Furthermore, we observed cell cycle arrest at G₀/G₁ phase as well as a block in G₂/M transition. In addition, combined treatment with the chemotherapeutic agents streptozocin and cisplatin augmented significantly the antiproliferative effects of those agents. Conclusion: Aurora kinase inhibition by ZM447439 seems to be a promising new therapeutic approach in GEP-NETs, which should be evaluated in further clinical trials.

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Introduction: Abnormal blood levels of various trace elements have not been consistently studied in the end-stage renal disease. The uremic patients treated by chronic dialysis lose one important route of trace elements excretion and are exposed to an artificial environment of the dialysis fluid. The process may lead to depletion of biologically essential substances or its accumulation in the patient. Thus, dialysis patients are at risk for both deficiency and accumulation of trace elements, depending on dietary intake, residual kidney function and removal by dialysis. Aim of the study:The present study was pointed to investigate possible existence of trace elements disturbances in uremic patients undergoing regular hemodialysis (HD) treatment and its relationship to some inflammatory markers.Material and methods: Blood samples of 56 patients on hemodialysis and 30 healthy controls were analysed twice in 3 months follow up period for quantitation of serum zinc, selenium and copper, serum albumins -transporters of the TE in the human body, and C-reactive protein (CRP) and interleukine-6 (Il-6) -two important markers of inflammation. Results:The study revealed that serum zinc and selenium concentrations in HD patients are distinctly decreased compared to that of healthy controls, and serum copper was found slightly abnormally increased. Significant negative correlation was found between Zn/CRP and Zn/Il-6 and positive one -between Cu/CRP and Cu/Il-6. Conclusion:Disturbances of trace elements are primarily the result of renal failure, but they may be further exacerbated by the dialysis procedure. Negative correlations between Zn and some inflammatory markers and positive -between them and Cu, probably suggest some influence of these TE on the inflammatory process in dialysis patients.

Introduction:We present a case of a 40-year-old woman with IVF gemellar pregnancy in the 28th week of gestation, with primary hyperparathyroidism which complicated the course of pregnancy causing acute pancreatitis and AKI, who was treated with CRRT and succesfully overcame a hypercalcemic crisis. Methods:Case report: On admission to the Obstetrics and gynecology clinic patient was somnolent, hypertensive, tachicardic, oliguric, respiratory stable with pretibial oedema. Laboratory data showed raised inflammatory markers, anemia, elevated serum amylases, urea, creatinine and hypokalemia. Abdominal ultrasound revealed an enlarged, voluminous pancreas, whereas chest radiograph showed a large left sided pleural effusion. An inital diagnosis of severe preeclampsia was determined, with suspected acute pancreatitis. Emergent cesarean delivery was performed. In the post partum period she was treated with isotonic saline infusions, antibiotic therapy (cephalosporins, carbapenems), antihypertensive drugs, anticonvulsants, antiedematous therapy with preventive doses of heparin. After two days she was transfered to the Intensive Care Unit. She was dyspnoic with compensated respiratory acidosis. Additional laboratory findings indicated high levels of serum lipases and severe hypercalcemia (total calcium: 3,99mmol/l, ionized calcium 2,47mmol/l, hyperphosphatemia 0,45mmol/l, high levels of parathyroid hormone 834pg/ml) and hypokalemia. CT of the chest and abdomen, revealed acute pancreatitis, bilateral pleural effusions and signs of AKI. Endocranial MR showed signs of brain edema. US of the thyroid and parathyroid gland identified a cystic formation with clear borders and intranodular vascularisation in the parenchyma of the lower left lobe, size 9x13x26mm, resembling an enlarged parathyreoid gland. Other causes of hypercalcemia were excluded. Results: Previous therapy was continued with the addition of hydration (rate of 200ml/h), proton -pump inhibitors, corticosteroids, bolus doses of furosemide and byphosphonates (calcitonin was unavailable). Two combined pre-dilution procedures were performed using heparin anticoagulation and normal calcium levels of 1,5mmol/l in the dialysate. Initially CVVHDF was started (Multifiltrate Kit 8 CVVHF 1000, surface 1,8m 2 ; flow dialysate 200-300ml/h; blood flow 180-100ml/h; dialytic fluid/substitute ratio was 1:1) and further changed to CVVH with continuous potassium substitution. After the first CRRT procedure, a decrease in calcium levels was noted, with tendency for further reduction, resulting in desired (total calcium 2,23mmol/l, ionized calcium levels 1.26mmol/l) and gradual normalisation of other laboratory findings. The patients state of consciousness improved, diuresis was established and complete hemodynamic stability was reached, after which, on the 12th day of treatment, she was transfered to the Clinic of endocrionology for further treatment. Conclusions: Combining CRRT modality with heparin anticoagulation and careful monitoring of electrolyte levels can contribute to adequate...