Molecular diagnostic tools for the detection of nodal micrometastases in prostate cancer patients undergoing radical prostatectomy with extended pelvic lymph node dissection: a prospective study

Background: Routine pathological examination can miss micro-metastatic tumor foci in the lymph nodes (LN) of patients with prostate cancer (PCa) that undergo radical prostatectomy and pelvic lymph node dissection (PLND). The aim of the present prospective study was to evaluate the impact of micrometastases assessed by serial section (SS), immunohistochemistry (IHC), and Real-time Polymerase Chain Reaction (RT-PCR) in patients undergoing radical prostatectomy with extended PLND. Materials and methods: 32 consecutive patients who underwent radical prostatectomy with extended PLND (obturator, internal/external and distal 2cm common iliac lymph-nodes (LN)) for intermediate (clinical T1c-T2 and PSA:10-20 ng/mL and clinical Gleason Score = 7) or high (clinical stage T3 or PSA>20 or clinical Gleason Score = 8-10) PCa were enrolled. The nodes were processed by the one uropathologist, both according to the routine pathological examination (analysis of the central section for 4 mm nodes or every 2 mm for LN>4 mm), which served as comparative method, both according to SS, IHC with antibodies against PSA and broad-spectrum Cytokeratins (BSCK), and quantitative RT-PCR targeting PSA, PSMA (PS Membrane Antigen), and Glucuronidase-S-Beta (GUSB) mRNA, that are over-expressed in prostatic cancer cells. Results: A total of 628 LN were analyzed, with a mean number of LN removed of 19.6 (SD = 7.2). Applying the routine pathological examination, 10 (31.2%) patients and 23 (3.9%) LN resulted positive for nodal involvement, with mean positive LN of 2.2 (SD = 1.4). After applying the SS and the molecular method of analysis (IHC and RT-PCR), micrometastases were found in 7 LN (SS showed micrometastases in 3 of them, IHC in 6 of them and RT-PCR in 7 of them); a total of 3 (9.3%) node-negative patients showed micrometastases at routine pathological examination (in 2 patients with RT-PCR and in 1 with IHC). Conclusions: The significance of micrometastases in PCa and the potential therapeutic role of PLND is not yet clarified, but the molecular analysis of the LN can detect a significant percentage of patients who harbor micro-metastatic PCa missed at routine pathological examination, and can enhance the accuracy of lymphadenectomy as a staging method.     

Telomerase activity,telomere length and hTERT DNA methylation in peripheral blood mononuclear cells from monozygotic twins with discordant smoking habits

Increased telomerase expression has been implicated in the pathogenesis of lung cancer and, since the primary cause of lung cancer is smoking, an association between telomerase reactivation and tobacco smoke has been proposed. In this work an investigation has been performed to assess the relationship between tobacco smoke exposure and telomerase activity (TA) in peripheral blood mononuclear cells of healthy smokers. The methylation status of the catalytic subunit of telomerase hTERT was concurrently investigated to assess the possible association between epigenetic modifications of hTERT and TA. Besides, the association between smoke and telomere length (TL) has been evaluated. Healthy monozygotic twins with discordant smoking habits were selected as study population to minimize inter‐individual differences because of demographic characteristics and genetic heterogeneity. Statistically significant higher values of TA and TL were observed in smokers compared to nonsmoker co‐twins. The multivariate analysis of data showed, besides smoking habits (P = 0.02), an influence of gender (P = 0.006) and BMI (P = 0.001) on TA and a borderline effect of gender (P = 0.05) on TL. DNA methylation analysis, focused on 100 CpG sites mapping in hTERT, highlighted nine CpG sites differentially methylated in smokers. When co‐twins were contrasted, selecting as variables the intra‐twin difference in TA and hTERT DNA methylation, a statistically significant inverse correlation (P = 0.003) was observed between TA and DNA methylation at the cg05521538 site. In conclusion, these results indicate an association of tobacco smoke with TA and TL and suggest a possible association between smoke‐induced epigenetic effects and TA in healthy smokers. Environ. Mol. Mutagen. 58:551–559, 2017. © 2017 Wiley Periodicals, Inc.     

Role of robotic-assisted pancreatic surgery:lessons learned from our initial experience

BACKGROUND:Minimally invasive surgery has achieved worldwide acceptance in various fields, however, pancreatic surgery remains one of the most challenging abdominal pro-cedures. In fact, the indication for robotic surgery in pancre-atic disease has been controversial. The present study aimed to assess the safety and feasibility of robotic pancreatic resec-tion. METHODS: We retrospectively reviewed our experience of robotic pancreatic resection done in Sanchinarro University Hospital. Clinicopathologic characteristics, and perioperative and postoperative outcomes were recorded and analyzed. RESULTS: From October 2010 to April 2016, 50 patients underwent robotic-assisted surgery for different pancreatic pathologies. All procedures were performed using the da Vinci robotic system. Of the 50 patients, 26 were male and 24 female. The average age of all patients was 62 years. Operative time was 370minutes. Among the procedures performed were 16 pancreaticoduodenectomies (PD), 23 distal pan-createctomies (DP), 11tumor enucleations (TE). The mean hospital stay was 17.6 days in PD group, 9.0 days in DP group and 8.4 days in TE group. Pancreatic fistula occurred in 10 cases (20%), 2 after PD, 3 after DP, and 5 after TE. Four pa-tients had postoperativetransfusion in PD group and one in DP group. Conversion to open laparotomy occurred in four patients (8%). No serious intraoperative complications were observed. CONCLUSIONS:From our early experience, robotic pancre-atic surgery is a safe and feasible procedure. Further experi-ence and follow-up are required to confirm the role of robotic approach in pancreatic surgery.     

Risk factors in transient osteoporosis: a retrospective study on 23 cases

The aim of this study is to verify the prevalence of risk factors for transient osteoporosis (TO) in a cohort of patients selected by strict diagnostic criteria. Retrospective observational cohort study on outpatients’ data. Inclusion criteria were: (1) acute onset of pain at a lower limb joint exacerbated by weight bearing; (2) no history of trauma, tumors, rheumatic diseases, or infection; (3) presence bone marrow edema on MRI in a weight bearing joint without signs of intraarticular lesions; (4) no hyperesthesia and/or allodynia and/or sweeting changes. The following risk factors were search for in all patients: (1) previous episode of TO; (2) disorders of bone metabolism; (3) cigarette smoke; (4) sudden lower limb overuse; (5) presence of osteoporosis/osteopenia. Twenty-three patients (8 females, 15 males, mean age 48.4 years) fulfilled the inclusion criteria. An average of 1.96 risk factors for TO was present in the cohort. The most frequent risk factor was overuse (in 15 patients, 65.2 %) and the second risk factor was bone metabolism disorders (in 10 patients, 43.5 %). Seven patients (30.4 %) were heavy smokers (more than 20 cigarettes per day) and seven patients showed a previous episode of TO. Six patients (26.1 % of the overall cohort but 60 % of those investigated with DEXA) resulted osteoporotic or osteopenic. Our results suggest there are risk factors that must be investigated in these patients. The presence of these risk factors might support the thesis that their disorder is tied to a decoupling between microdamage accumulation and self-reparative ability of bone tissue. The identification of risk factors with a precise diagnostic pathway can accelerate the diagnostic process and reduce recurrences.