Beta Elemene induces cytotoxic effects in FLT3 ITD-mutated acute myeloid leukemia by modulating apoptosis.

OBJECTIVE: ß-Elemene, a sesquiterpene with a broad anti-cancer spectrum, is particularly effective against drug-resistant and complex tumors. It can also be efficient against FLT3-expressed acute myeloid leukemia. This research aims to determine whether ß-Elemene has cytotoxic effects on FLT3 ITD-mutated AML cells. MATERIALS AND METHODS: Cytotoxicity, cell morphology, mRNA analysis with apoptotic markers, and analysis of 43 distinct protein markers related to cell death, survival, and resistance were all performed to elucidate its mechanism. Additionally, in order to understand how ß-Elemene and FLT3 interact, molecular docking, molecular dynamics simulations, and computational ADME investigations were performed. RESULTS: ß-Elemene exhibited cytotoxic activity against FLT3-mutated MV4-11 and FLT3 wild-type THP-1 cells, with an IC50 of around 25 µg/ml. The molecular studies revealed that ß-Elemene inhibited cell proliferation by inducing p53, and the involvement of p21, p27, HTRA, and HSPs were also demonstrated. The interactive inhibition in proliferation was confirmed via molecular docking and dynamics analyses. ß-Elemene occupied the FLT3 enzymatic pocket with good stability at the FLT3 active site. CONCLUSIONS: We concluded from our observations that ß-Elemene causes cell death in ITD mutant AML cells, together with the effects of stress factors and inhibiting cell division.

Personalized risk prediction of postoperative cognitive impairment - rationale for the EU-funded BioCog project.

Postoperative cognitive impairment is among the most common medical complications associated with surgical interventions - particularly in elderly patients. In our aging society, it is an urgent medical need to determine preoperative individual risk prediction to allow more accurate cost-benefit decisions prior to elective surgeries. So far, risk prediction is mainly based on clinical parameters. However, these parameters only give a rough estimate of the individual risk. At present, there are no molecular or neuroimaging biomarkers available to improve risk prediction and little is known about the etiology and pathophysiology of this clinical condition. In this short review, we summarize the current state of knowledge and briefly present the recently started BioCog project (Biomarker Development for Postoperative Cognitive Impairment in the Elderly), which is funded by the European Union. It is the goal of this research and development (R&D) project, which involves academic and industry partners throughout Europe, to deliver a multivariate algorithm based on clinical assessments as well as molecular and neuroimaging biomarkers to overcome the currently unsatisfying situation.

Clinical Significance of Intraoperative Frozen Section Analysis of Pancreatic Cancer Surgical Margin at the Time of Pancreaticoduodenectomy.

BACKGROUND: Today, in the management of pancreas cancers, achieving an R0 resection is one of the most powerful independent predictors of long-term survival. The aim of this study is to assess the clinical significance of intraoperative frozen section analysis of the pancreatic surgical margin for pancreatic cancer during pancreaticoduodemectomy. MATERIAL AND METHODS: We conducted a retrospective analysis of prospectively collected data of 37 patients who were operated for pancreatic head cancer and who were evaluated for surgical margin by frozen section analysis intraoperatively, between September 2013 and August 2014 in our center. The intraoperative biopsy reports were compared with final pathological reports. RESULTS: The mean age of the patients was 64.55(19-82) years (range), the mean tumor size was 3.96(1.16-6.25) cm (range) and the mean harvested lymph node number was 18.52(9-45) (range). In the intraoperative frozen section, one patient was positive for surgical margin (%2.7) who underwent total or complementary pancreatectomy. CONCLUSION: To secure a tumor-free margin by frozen section, intraoperatively, may increase R0 resection rate in pancreas cancers. The preoperative estimation of tumor margin by endoscopic ultrasonography, computerized tomography or magnetic resonance imaging mostly correlate with intraoperative findings, however in suspected cases intraoperative frozen section for margin determination should be performed.

Use of ductoscopy as an additional diagnostic method and its applications in nipple discharge.

AIM: About 1/10 of the patients apply to breast clinics with the complaint of nipple discharge (ND). Surgery is the most frequently preferred treatment method in case of suspicious ND. The contribution of ductoscopy to identify the patients who are candidates for surgery was evaluated and its role to limit the surgery was assessed. METHODS: From November 2005 to December 2010 430 patients with ND were assessed by 456 ductoscopic investigations and the results were analyzed. Complete ductoscopic evaluation was achieved in 84% of cases and 28 patients were offered surgery but did not accept (N.=355). Patients with bloody or serous discharges from a single duct were investigated by ductoscopy under local anesthesia as an office procedure. The patients were grouped according to discharge characteristics and the ductoscopic diagnoses. RESULTS: A total of 223 patients had all three criteria of pathologic ND (PND: single duct, spontaneous and bloody/serous discharge). Twenty-two potential neoplastic or malignant lesion (PNML) and 79 papillomatous lesion (solitary or multiple papilloma) were identified. In 132 patients with just two of the PND criteria, 5 PNML and 18 papillomatous lesions were identified. Twenty-three patients with solitary papilloma that were removed by ductoscopic papillomectomy (DP) are followed up without surgery. CONCLUSION: Ductoscopy helped to identify the patients who required surgical treatment and decreased the number of operations. DP was successfully performed in select group of patients who otherwise would have required surgical resection. Patients with normal ductoscopy findings and patients who were treated with DP successfully can be followed up without the need of surgery.

Biochemical characterization of the vanilloid receptor 1 expressed in a dorsal root ganglia derived cell line.

The vanilloid receptor VR1 is an ion channel predominantly expressed by primary sensory neurons involved in nociception. Here we describe its biochemical properties and assess the subcellular localization, the glycosylation state and the quaternary structure of VR1 expressed in HEK293 cells and in the DRG-derived cell line F-11 (N18TG2 mouse neuroblastoma x rat dorsal root ganglia, hybridoma). VR1 was found to be glycosylated in both cell types. Of the five potential N-glycosylation sites, the predicted transient receptor potential channel-like transmembrane folding proposes N604 is localized extracellularly. We used site-directed mutagenesis to mutate the Asn at position 604 to Thr. This mutated VR1 was not glycosylated, confirming the extracellular location of N604 and its role as the exclusive site of glycosylation of the VR1 protein. VR1 occured in high molecular mass complexes as assessed by blue native PAGE. In the presence of limited amounts of SDS dimers, trimers and tetramers of VR1 were observed, consistent with the predicted tetrameric quaternary structure of the receptor. Cross-linking with dimethyladipimidate yielded almost exclusively dimers. Whereas VR1 localized both to the plasma membrane and to intracellular membranes in HEK293 cells, it localized predominantly to the plasma membrane in F-11 cells. Using confocal laserscanning microscopy, we observed an enrichment of anti-VR1 immunoreactivity in neurite-like structures of F-11 cells. In the light of conflicting literature data on biochemical characteristics of VR1, our data suggest that dorsal root ganglion-derived F-11 cells provide a powerful experimental system for the study of VR1 biochemistry.

Intraoperative bowel irrigation improves anastomotic collagen metabolism in the left-sided colonic obstruction but not covering colostomy.

This study investigated the effects of intraoperative colonic irrigation and proximal diverting end colostomy after segmental bowel resection in experimental left-colonic obstruction on anastomotic healing. Simple obstruction of descending colon was performed in male Sprague-Dawley rats. After 24 h we performed segmental colonic resection and anastomosis in the control group (n = 15); resection, anastomosis, and covering colostomy in the colostomy group (n = 14); resection and anastomosis after antegrade colonic lavage through cecum by using isotonic saline solution in the irrigation group (n = 13). In rats that were killed 7 days later anastomotic dehiscence and bursting pressure and tissue hydroxyproline concentration at the anastomosis were measured. No significant differences were observed between groups in terms of anastomotic dehiscence, bursting site, or pressure. The hydroxyproline concentration was significantly higher in the irrigation group than the control group (P = 0.025) and the colostomy group (P = 0.029), but no difference was noted between the control group and the colostomy group. These findings suggest that intraoperative antegrade colonic irrigation in the acute left-sided colonic obstruction positively affects collagen metabolism at the anastomotic site; if the anastomosis is performed without bowel cleansing, covering colostomy does not improve collagen metabolism.

The Fanconi anaemia group G gene FANCG is identical with XRCC9.

Fanconi anemia (FA) is an autosomal recessive disease with diverse clinical symptoms including developmental anomalies, bone marrow failure and early occurrence of malignancies. In addition to spontaneous chromosome instability, FA cells exhibit cell cycle disturbances and hypersensitivity to cross-linking agents. Eight complementation groups (A-H) have been distinguished, each group possibly representing a distinct FA gene. The genes mutated in patients of complementation groups A (FANCA; refs 4,5) and C (FANCC; ref. 6) have been identified, and FANCD has been mapped to chromosome band 3p22-26 (ref. 7). An additional FA gene has recently been mapped to chromosome 9p (ref. 8). Here we report the identification of the gene mutated in group G, FANCG, on the basis of complementation of an FA-G cell line and the presence of pathogenic mutations in four FA-G patients. We identified the gene as human XRCC9, a gene which has been shown to complement the MMC-sensitive Chinese hamster mutant UV40, and is suspected to be involved in DNA post-replication repair or cell cycle checkpoint control. The gene is localized to chromosome band 9p13 (ref. 9), corresponding with a known localization of an FA gene.

Identification of a HeLa mRNA fraction which corrects the mitomycin C sensitivity of irs1 cells.

The hamster cell mutant irs1 is defective in its response to DNA lesions caused by a variety of mutagens, particularly cross-linking agents. These cells have been assigned to complementation group 2 of X-ray-sensitive mutants and the mutated gene is called XRCC2(X-ray repair cross complementing). We have identified, by microinjection, a human mRNA fraction which can transiently correct the sensitivity of these cells to cross-linking agents. This fraction contains mRNAs of 3.5 kb (+/- 0.25) including, therefore, the transcript of the XRCC2 gene.