Genetic and epigenetic changes in primary metastatic and nonmetastatic colorectal cancer

Colorectal cancer (CRC) develops as multistep process, which involves genetic and epigenetic alterations. K-Ras, p53 and B-Raf mutations and RASSF1A, E-Cadherin and p16INK4A promoter methylation were investigated in 202 CRCs with and without lymph node and/or liver metastasis, to assess whether gene abnormalities are related to a metastogenic phenotype. K-Ras, B-Raf and p53 mutations were detected in 27, 3 and 32% of the cases, with K-Ras mutations significantly associated with metastatic tumour (P=0.019). RASSF1A, E-Cadherin and p16INK4A methylation was documented in 20, 44 and 33% of the cases with p16INK4A significantly associated with metastatic tumours (P=0.001). Overall, out of 202 tumours, 34 (17%) did not show any molecular change, 125 (62%) had one or two and 43 (21%) three or more. Primary but yet metastatic CRCs were prevalent in the latter group (P=0.023) where the most frequent combination was one genetic (K-Ras in particular) and two epigenetic alterations. In conclusion, this analysis provided to detect some molecular differences between primary metastatic and nonmetastatic CRCs, with K-Ras and p16INK4A statistically altered in metastatic tumours; particular gene combinations, such as coincidental K-Ras mutation with two methylated genes are associated to a metastogenic phenotype

P-162ROBOTIC APPROACH IN THE TREATMENT OF STAGE III LUNG CANCER: A RETROSPECTIVE MULTICENTRE ANALYSIS OF PERIOPERATIVE AND ONCOLOGICAL RESULTS

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NUTRITIONAL AND DIGESTIVE EFFECTS OF GASTRECTOMY FOR GASTRIC CANCER

Background: Gastrectomy often leads to malnutrition. Objective: The aim of this study was to analyze nutritional and digestive effects of gastrectomy for cancer. Patients and methods: Gastrectomized patients were studied by nutritional assessment including a weekly nutritional diary exploring digestive symptoms. Results: Thirty-two patients were analyzed after a mean follow-up of 41.8 months. The mean percentage of weight loss was 12.9% \ub1 13.5%. After total gastrectomy, mean weight loss was 22% \ub1 1.2%, against 7.4% \ub111.9% for subtotal gastrectomy (p = 0.002). Moreover, advancing age was related to weight loss (p = 0.02), with a peak around 70 years. The most frequent postprandial symptoms were abdominal swelling (62%) and early satiety (59%). Finally, findings of the present study imply that overm a long follow-up, there are no specific intolerances related to gastrectomy. Conclusions: Patients who have undergone a total gastrectomy and elderly gastrectomized patients are at risk of malnutrition and need postoperative nutritional support

Analysis of epidermal growth factor receptor expression as a predictive factor for response to gefitinib (‘Iressa’, ZD1839) in non-small-cell lung cancer

Gefitinib ('Iressa', ZD1839) is an orally active epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor that has demonstrated antitumour activity and favourable tolerability in Phase II studies. We investigated whether EGFR expression levels could predict for response to gefitinib in patients with advanced non-small-cell lung cancer (NSCLC), who received gefitinib (250 mg day(-1)) as part of a worldwide compassionate-use programme. Tissue samples were analysed by immunohistochemistry to assess membrane EGFR immunoreactivity. Of 147 patients enrolled in our institution, 50 patients were evaluable for assessment of both clinical response and EGFR expression. The objective tumour response rate was 10% and disease control was achieved in 50% of patients. Although high EGFR expression was more common in squamous-cell carcinomas than adenocarcinomas, all objective responses were observed in patients with adenocarcinoma. Response and disease control with gefitinib were not associated with high EGFR expression. Overall, median survival was 4 months, and the 1-year survival rate was 18%. Strong EGFR staining correlated with shorter survival time for all patients. Gefitinib demonstrated promising clinical activity in this group of patients with NSCLC. These results have also shown that EGFR expression is not a significant predictive factor for response to gefitinib

Time-dependent ROC curves in genetically determined subgroups

The ultimate goal of genetic association studies is to identify and map the gene(s) responsible for a given disease. This paper discusses a new and simple statistical method for detecting a genetic association, based on time-dependent Receiver Operating Characteristic (ROC) curves. This method resorts to the Heagerty approach based on Bayes theorem, and uses the Kaplan-Meier or the Akritas estimator. An application to the real problem of examining possible interaction between glycaemia and a risk \u201cgenotype\u201d on survival is presented using the Framingham database. Analysis assessed area of chromosome 1 (from 192 to 233 cM) and evaluated the role of fasting blood glucose on survival, at 4 and 8 years of follow-up, according to the presence/absence of allele 242 (marker 23). The allele 242 showed ability in predicting survival. Kaplan-Meier and Akritas estimators provided comparable results. At 4 years of follow-up, area (SD) under ROC curve, in absence of allele 242, was 0.85 (0.09) and 0.81 (0.07) using respectively the Kaplan-Meier or Akritas estimator. The ability showed by the time-dependent ROC curves in real data suggests that the proposed method may be valuable to detect difference in genetic subgroups. Further studies need to better clarify the usefulness of this method in other real applications

Cracking spaces in Hashimoto Thyroiditis are lymphatic and prelymphatic vessels : a gift of immunohistochemistry for the centenary of Hashimoto's description

A century ago Hashimoto described the histologic hallmarks of struma lymphomatosa: (1) lymphoid follicles, (2) changes in the epithelial cells, (3) formation of connective tissue, and (4) diffuse round cell infiltration. He also showed some cracking spaces close to lymphoid follicles resembling vessels. The aim of this study was to investigate the possible lymphatic nature of these spaces and their prevalence in non-neoplastic thyroid tissue. Ten cases of Hashimoto thyroiditis (HT), 5 of Basedow-Graves disease (BG), and 5 of normal thyroid tissue (NT) were selected. Tissues were stained with hematoxylin and eosin, CD31 (Dako), and D2-40 (Dako) stains. Cracking spaces staining positive for CD31 and D2-40 stains were considered as lymphatic vessels. Site, distribution, intravascular valves and lymphocytes, perivascular lymphoid aggregates, and number and surface of lymphatic vessels were evaluated using a computer-assisted digital videocamera microscope (Nikon digital sigh, DS-2Mv). The number of lymphatic vessels increased from NT [3 (range, 2 to 13)] to BG [8 (range, 5 to 9)] to HT [12.5 (range, 10 to 16)]. A significant statistical difference was observed within the group (P=0.003): HT differed from NT (P=0.016) and BG (P=0.002). The area of lymphatic vessels increased from NT [0.01 mm (range, 0.01 to 0.12 mm2)] to BG [0.03 mm2 (range, 0.01 to 0.19 mm2)] to HT [0.03 mm (range, 0.01 to 0.6 mm2)]. A significant statistical difference was observed among the groups (P=0.001): NT differed from HT (P<0.001) and from BG (P<0.001). Lymphatic vessels showed valves, perivascular lymphoid aggregates, and intravascular lymphocytes. The cracking spaces shown by Hashimoto are mainly lymphatic vessels and represent a characteristic feature of autoimmune thyroid diseases

Five years (2004-2009) of a restrictive law-regulating ART in Italy significantly reduced delivery rate: analysis of 10 706 cycles

Study question: Law 40/2004 enacted by the Italian Parliament established a long list of restrictions for ART procedures. We investigated the differences in the delivery rate of all ART cycles performed before and after the introduction of the law. Summary answer: This large and extensive comparative analysis of ART outcomes prior and after the introduction of the Law 40 revealed a significant reduction in pregnancy and delivery rates per cycle, independent of age or other clinical variables, once the law went into effect. What is known and what this paper adds: Several studies have been published on the effect of Law 40/2004 on ART outcomes, some authors demonstrating a negative impact of the law also in relation to specific aetiologies of infertility, other authors showing opposite conclusions. Our study, the first based on the analysis of a so large number of cycles, clearly demonstrated a negative effect of Law 40 restrictions on ART procedures, as an independent factor responsible for the reduced delivery rate of IVF cycles. Design: Retrospective clinical study Partecipants and setting: 10,706 cycles were analysed, 3,808 performed before and 6,898 cycles after the application of the Law. An intention to treat statistical analysis was performed to detect pregnancy and delivery rates (pregnancies ≥ 24 weeks) per started cycle. A p value <0.05 was considered statistically significant. We analysed different outcomes: differences in fertilization, pregnancy, and delivery rate, multiple pregnancies and miscarriage rates between the two time periods. Main results and the role of chance: The delivery rate for started cycle was 20% before and 16.0% after the introduction of the Law representing a 25% reduction (p<0.001). The multivariate analysis, corrected by age, basal FSH level, number of retrieved oocytes and total progressive motile count, showed a 16% lower delivery rate (OR 0.84 CI 0.75-0.94) and confirmed the effect of the Law as an independent factor responsible for the reduced chances of a successful outcome

One step further: randomised single-centre trial comparing the direct and afterload techniques of embryo transfer

STUDY QUESTION: What are the differences in ease of use between two different embryo transfer (ET) techniques: the preload direct approach and the afterload approach. SUMMARY ANSWER: The afterload technique seems to reduce the rate of difficult ETs. WHAT IS KNOWN ALREADY: Numerous published trials now document that the ET procedure has an impact on pregnancy and delivery rates after IVF. Difficult transfers should be avoided, as they reduce implantation and pregnancy rates. Preload direct ETs with soft catheters under ultrasound guidance is currently considered the best procedure. However, when using soft catheters, it is not known which technique is preferable or which one should be implemented to reduce the operator factor. STUDY DESIGN, SIZE, DURATION: This prospective randomised unblinded controlled clinical trial, included 352 ultrasound-guided ETs assigned to either direct ET or after-load ET, between September 2017 and March 2019. The sample size was calculated based on the historical rate of difficult ETs encountered between 2014 and 2015 with a direct ET procedure. PARTICIPANTS/MATERIALS, SETTING, METHODS: The inclusion criteria were women 18-38 years old, with BMI between 18 and 28, receiving a single-thawed blastocyst transfer. The exclusion criteria were use of testicular sperm and preimplantation genetic testing (PGT) cycles. The primary outcome was the rate of difficult or suboptimal transfers defined as: advancement of the outer sheath (specific for the direct transfer), multiple attempts, use of force, required manipulation, use of a stylet or tenaculum, dilatation, or use of a different catheter. The secondary outcome was clinical pregnancy rate. MAIN RESULTS AND THE ROLE OF CHANCE: A total of 352 frozen ETs were randomised, with 176 patients in each group. The two arms were homogeneous for female and male age, female BMI, duration of infertility, secondary infertility, previous deliveries or miscarriages, myomas, previous surgery to the uterine cavity, cycle day at ovulation trigger, freeze all cycles, first transfers, indication for treatment, endometrial preparation protocol and duration, endometrial thickness, and blastocyst grade at vitrification. Across the entire population, 85 (24.1%) ETs were defined as difficult. The rate of difficult transfers was significantly higher in the direct ET group than in the afterload group: 68 (38.6%) versus 17 (9.7%), respectively (OR 0.17, 95% CI 0.09-0.30, P < 0.001). The mean percentage in the rate of difficult transfers per operator was 22.5% (SD +/- 14.5%), of which 36.1% (SD +/- 23.4%) were in the direct group compared with 8.6% (+/- 8.2%) in the afterload group (P < 0.001). The difficult transfer rate among operators varied from 0 to 43.8% (0-77.8% in the direct group and 0 to 25.0% in the afterload group). The clinical pregnancy rates (42.0% vs 48.3%, P = 0.239 in the direct and afterload groups, respectively) were not significantly different between the groups. LIMITATIONS, REASONS FOR CAUTION: There were 18 experienced operators who participated in the trial. Conclusions about the pregnancy rate should not be generalised, since the sample analysis was not performed on this outcome and, although clinically relevant, the difference was not significantly different. WIDER IMPLICATIONS OF THE FINDINGS: The rate of difficult transfers was significantly higher in the direct ET group compared with the afterload ET group, although a wide variation was observed among operators. Further studies regarding the association between transfer technique and ART outcomes are required. STUDY FUNDING/COMPETING INTEREST(S): No specific funding was sought and there are no competing interests

Dose-related effects of metformin on acid&#8211;base balance and renal function in patients with diabetes who develop acute renal failure : a cross-sectional study

Aims: The role of metformin in the development of lactic acidosis (LA) in the setting of acute renal failure (ARF) is debated. Moreover, recent experiments suggested that metformin can also be nephrotoxic, but little clinical data exist about this topic. We sought to investigate these possible associations in a large cohort of patients with diabetes who developed ARF. Methods: We analyzed data from patients with diabetes admitted to our emergency department between 2007 and 2011 with ARF and a previously normal renal function (n&nbsp;=&nbsp;126). We compared acid\u2013base balance and renal function of patients taking metformin (n&nbsp;=&nbsp;74) with patients not taking it (n&nbsp;=&nbsp;52). Results: Patients taking metformin had decreased pH (7.31&nbsp;\ub1&nbsp;0.16 vs 7.39&nbsp;\ub1&nbsp;0.11, p&nbsp;=&nbsp;0.008) and higher lactates (4.54&nbsp;\ub1&nbsp;4.30 vs 1.71&nbsp;\ub1&nbsp;1.14&nbsp;mmol/L, p&nbsp;&lt;&nbsp;0.001). Both acidosis (pH&nbsp;&lt;&nbsp;7.35) and LA (lactates &gt;5&nbsp;mmol/L and pH&nbsp;&lt;&nbsp;7.35) were more frequently observed in this group (p&nbsp;=&nbsp;0.0491 and p&nbsp;&lt;&nbsp;0.001, respectively). Multivariate analysis ruled out the role of some possible confounders, especially decreased renal function. The influence of metformin on pH and lactates grew significantly with higher doses of the drug (p&nbsp;=&nbsp;0.259 and p&nbsp;=&nbsp;0.092 for &lt;1&nbsp;g/day, p&nbsp;=&nbsp;0.289 and p&nbsp;&lt;&nbsp;0.001 for 1\u20132&nbsp;g/day, p&nbsp;=&nbsp;0.009 and p&nbsp;&lt;&nbsp;0.001 for 2\u20133&nbsp;g/day, for pH and lactates, respectively). Metformin influenced creatinine levels in a dose-related manner as well (p&nbsp;=&nbsp;0.925 for &lt;1&nbsp;g/day, p&nbsp;=&nbsp;0.033 for 1\u20132&nbsp;g/day, p&nbsp;&lt;&nbsp;0.001 for 2\u20133&nbsp;g/day). Conclusions: In patients with diabetes who were admitted to our emergency department with ARF, the use of metformin was associated in a dose-related fashion with both LA and worse renal function