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Purpose: A recent study reported that 5-fluorouracil (5-FU)-based chemotherapy is less effective in treating patients with advanced colorectal cancer demonstrating hypermethylation of the TFAP2E gene. The aim of our study was to confirm and validate these findings in large, uniformly treated, well-characterized patient cohorts.Experimental Design: Two cohorts of 783 patients with colorectal cancer: 532 from a population-based, multicenter cohort (EPICOLON I) and 251 patients from a clinic-based trial were used to study the effectiveness of TFAP2E methylation and expression as a predictor of response of colorectal cancer patients to 5-FU–based chemotherapy. DNA methylation status of the TFAP2E gene in patients with colorectal cancer was assessed by quantitative bisulfite pyrosequencing analysis. IHC analysis of the TFAP2E protein expression was also performed.Results: Correlation between TFAP2E methylation status and IHC staining was performed in 607 colorectal cancer samples. Among 357 hypermethylated tumors, only 141 (39.6%) exhibited loss of protein expression. Survival was not affected by TFAP2E hypermethylation in stage IV patients [HR, 1.21; 95% confidence interval (CI), 0.79–1.87; log-rank P = 0.6]. In stage II–III cases, disease-free survival was not influenced by TFAP2E hypermethylation status in 5-FU–treated (HR, 0.91; 95% CI, 0.52–1.59; log-rank P = 0.9) as well as in nontreated patients (HR, 0.88; 95% CI, 0.5–1.54; log-rank P = 0.7).Conclusions: TFAP2E hypermethylation does not correlate with loss of its protein expression. Our large, systematic, and comprehensive study indicates that TFAP2E methylation and expression may not play a major role in predicting response to 5-FU–based chemotherapy in patients with colorectal cancer. Clin Cancer Res; 24(12); 2820–7. ©2018 AACR.

Supplementary Figure 4. Epicolon I. Overall Survival Stage IV Patients: TFAP2E IHC Expression. Disease Free Survival Stage II and III Patients: TFAP2E IHC Expression. (A) Overall survival of patients with stage IV disease, according to TFAP2E expression status comparing staining scores 0 & 1 (Low expression) vs staining scores 2 & 3 (High expression;). (B) Overall survival of patients that received or did not receive (C) 5-FU based chemotherapy according to TFAP2E expression status. (D) Disease Free Survival (DFS) of patients with stage II and III CRC, according to TFAP2E expression status comparing staining scores 0 & 1 (Low expression) vs staining scores 2 & 3 (High expression) (E) DFS of stages II+III CRC patients that received or did not receive (F)) adjuvant chemotherapy according to TFAP2E expression status.

Supplementary Figure 3. Correlation between TFAP2E intron 3 methylation and IHC expression. When methylation was analysed as a continuous variable using linear regression analysis, we observed a trend towards negative correlation with IHC protein expression; however, this correlation was significant only when combining lack of and low TFAP2E expression (IHC scores 0 & 1) versus strong TFAP2E staining (IHC scores 2 & 3) (p= 0.015). Linear regression analysis comparing TFAP2E methylation and expression in CRC tissues (N=314) using IHC results by binary scoring system: positive/negative (A) staining scores 0 & 1 vs 2 & 3(B) and the staining intensity scale 0 to 3 (C). Correlation and performance measures between TFAP2E methylation and expression in CRC tissues using IHC and methylation qualitative results (staining scores 0 & 1 vs 2 & 3; (D).

Purpose: The present study aimed to determine the prevalence of MUTYH mutations in patients with multiple colonic polyps and to explore the best strategy for diagnosing MUTYH-associated polyposis (MAP) in these patients.Experimental Design: This study included 405 patients with at least 10 colonic polyps each. All cases were genetically tested for the three most frequent MUTYH mutations. Whole-gene analysis was performed in heterozygous patients and in 216 patients lacking the three most frequent mutations. Polyps from 56 patients were analyzed for the KRAS-Gly12Cys and BRAF V600E somatic mutations.Results: Twenty-seven (6.7%) patients were diagnosed with MAP, of which 40.8% showed serrated polyps. The sensitivity of studying only the three common variants was 74.1%. Of 216 patients without any monoallelic mutation in common variants, whole-gene analysis revealed biallelic pathogenic mutation in only one. G396D mutation was associated with serrated lesions and older age at diagnosis. There was a strong association between germinal MUTYH mutation and KRAS Gly12Cys somatic mutation in polyps. BRAF V600E mutation was found in 74% of serrated polyps in MUTYH-negative patients and in none of the polyps of MAP patients.Conclusions: We observed a low frequency of MUTYH mutations among patients with multiple adenomatous and serrated polyps. The MAP phenotype frequently included patients with serrated polyps, especially when G396D mutation was involved. Our results show that somatic molecular markers of polyps can be useful in identifying MAP cases and support the need for the complete MUTYH gene analysis only in patients heterozygous for recurrent variants. Clin Cancer Res; 20(5); 1158–68. ©2014 AACR.

CCR Translation for This Article from Prevalence and Characteristics of MUTYH-Associated Polyposis in Patients with Multiple Adenomatous and Serrated Polyps

Supplementary data Supplementary Methods Supplementary Table 1. Characteristics of Epicolon I patients (n=532) Supplementary Table 2. Characteristics of Clinical cohort patients (n=251) Supplementary Table 3. Primer sequences Supplementary Figure 1. Schematic representation of TFAP2E gene. The gene encoding transcription factor AP-2 epsilon (TFAP2E) is located on chromosome 1p34. In the schematic overview, exons are represented by blue boxes and introns with blue lines. The TFAP2E gene has two cytosine-phosphate-guanine (CpG) islands underscoring the potential of gene expression by means of CpG hypermethylation (green boxes). We used two quantitative methods, QMSP and pyrosequencing, covering both CpG islands in order to correlate methylation levels at these two locations. Supplementary Figure 2. Methylation analysis of TFAP2E CpG islands in CRC tissues. Calculation of cutoff value of TFAP2E Intron 3 methylation. Results comparing methylation of the TFAP2E CpG1 (Promoter/Exon 1) and CpG2 (Intron 3) in CRC tissues (N=50) by (A) Bisulfite pyrosequencing (% Methylation) (B-C) Bisulfite pyrosequencing results of TFAP2E Intron 3 methylation in normal colon mucosa from healthy subjects N-N (N=21); and CRC patients-matched tumor-and non-tumor samples N-C (N=41);(N=41). (D) Whitin-subject Receiver Operating Characteristics (ROC) analysis performed on measured TFAP2E % (percentage of methylation) values of patient-matched tumor- (N-N; N=21) and non-tumor (N-C; N=41) samples, respectively. The area under the ROC-curve was calculated as 0.73 (95% CI:0.62-0.82), indicating that the probability of observing a higher TFAP2E value in a tumor-sample of a patient compared to its related non-tumor sample is about 73%. Supplementary Figure 3. Correlation between TFAP2E intron 3 methylation and IHC expression. When methylation was analysed as a continuous variable using linear regression analysis, we observed a trend towards negative correlation with IHC protein expression; however, this correlation was significant only when combining lack of and low TFAP2E expression (IHC scores 0 & 1) versus strong TFAP2E staining (IHC scores 2 & 3) (p= 0.015). Linear regression analysis comparing TFAP2E methylation and expression in CRC tissues (N=314) using IHC results by binary scoring system: positive/negative (A) staining scores 0 & 1 vs 2 & 3(B) and the staining intensity scale 0 to 3 (C). Correlation and performance measures between TFAP2E methylation and expression in CRC tissues using IHC and methylation qualitative results (staining scores 0 & 1 vs 2 & 3; (D). Supplementary Figure 1. Schematic representation of TFAP2E gene. The gene encoding transcription factor AP-2 epsilon (TFAP2E) is located on chromosome 1p34. In the schematic overview, exons are represented by blue boxes and introns with blue lines. The TFAP2E gene has two cytosine-phosphate-guanine (CpG) islands underscoring the potential of gene expression by means of CpG hypermethylation (green boxes). We used two quantitative methods, QMSP and pyrosequencing, covering both CpG islands in order to correlate methylation levels at these two locations. Supplementary Figure 2. Methylation analysis of TFAP2E CpG islands in CRC tissues. Calculation of cutoff value of TFAP2E Intron 3 methylation. Results comparing methylation of the TFAP2E CpG1 (Promoter/Exon 1) and CpG2 (Intron 3) in CRC tissues (N=50) by (A) Bisulfite pyrosequencing (% Methylation) (B-C) Bisulfite pyrosequencing results of TFAP2E Intron 3 methylation in normal colon mucosa from healthy subjects N-N (N=21); and CRC patients-matched tumor-and non-tumor samples N-C (N=41);(N=41). (D) Whitin-subject Receiver Operating Characteristics (ROC) analysis performed on measured TFAP2E % (percentage of methylation) values of patient-matched tumor- (N-N; N=21) and non-tumor (N-C; N=41) samples, respectively. The area under the ROC-curve was calculated as 0.73 (95% CI:0.62-0.82), indicating that the probability of observing a higher TFAP2E value in a tumor-sample of a patient compared to its related non-tumor sample is about 73%. Supplementary Figure 3. Correlation between TFAP2E intron 3 methylation and IHC expression. When methylation was analysed as a continuous variable using linear regression analysis, we observed a trend towards negative correlation with IHC protein expression; however, this correlation was significant only when combining lack of and low TFAP2E expression (IHC scores 0 & 1) versus strong TFAP2E staining (IHC scores 2 & 3) (p= 0.015). Linear regression analysis comparing TFAP2E methylation and expression in CRC tissues (N=314) using IHC results by binary scoring system: positive/negative (A) staining scores 0 & 1 vs 2 & 3(B) and the staining intensity scale 0 to 3 (C). Correlation and performance measures between TFAP2E methylation and expression in CRC tissues using IHC and methylation qualitative results (staining scores 0 & 1 vs 2 & 3; (D). Supplementary Figure 4. Epicolon I. Overall Survival Stage IV Patients: TFAP2E IHC Expression. Disease Free Survival Stage II and III Patients: TFAP2E IHC Expression. (A) Overall survival of patients with stage IV disease, according to TFAP2E expression status comparing staining scores 0 & 1 (Low expression) vs staining scores 2 & 3 (High expression;). (B) Overall survival of patients that received or did not receive (C) 5-FU based chemotherapy according to TFAP2E expression status. (D) Disease Free Survival (DFS) of patients with stage II and III CRC, according to TFAP2E expression status comparing staining scores 0 & 1 (Low expression) vs staining scores 2 & 3 (High expression) (E) DFS of stages II+III CRC patients that received or did not receive (F)) adjuvant chemotherapy according to TFAP2E expression status.

Supplementary Figure 1. Schematic representation of TFAP2E gene. The gene encoding transcription factor AP-2 epsilon (TFAP2E) is located on chromosome 1p34. In the schematic overview, exons are represented by blue boxes and introns with blue lines. The TFAP2E gene has two cytosine-phosphate-guanine (CpG) islands underscoring the potential of gene expression by means of CpG hypermethylation (green boxes). We used two quantitative methods, QMSP and pyrosequencing, covering both CpG islands in order to correlate methylation levels at these two locations.

Supplementary Figure 2. Methylation analysis of TFAP2E CpG islands in CRC tissues. Calculation of cutoff value of TFAP2E Intron 3 methylation. Results comparing methylation of the TFAP2E CpG1 (Promoter/Exon 1) and CpG2 (Intron 3) in CRC tissues (N=50) by (A) Bisulfite pyrosequencing (% Methylation) (B-C) Bisulfite pyrosequencing results of TFAP2E Intron 3 methylation in normal colon mucosa from healthy subjects N-N (N=21); and CRC patients-matched tumor-and non-tumor samples N-C (N=41);(N=41). (D) Whitin-subject Receiver Operating Characteristics (ROC) analysis performed on measured TFAP2E % (percentage of methylation) values of patient-matched tumor- (N-N; N=21) and non-tumor (N-C; N=41) samples, respectively. The area under the ROC-curve was calculated as 0.73 (95% CI:0.62-0.82), indicating that the probability of observing a higher TFAP2E value in a tumor-sample of a patient compared to its related non-tumor sample is about 73%.

The implementation of strategies to mitigate possible cases of COVID-19 were addressed at the University of Alicante for the safe reopening of the 2020/2021 academic year. To discover the prevalence of immunity against SARS-CoV-2, a study was designed using a rapid immunoassay test (carried out between 6 and 22 July 2020), and in addition a cross-sectional survey was conducted on risk factors, symptoms, predisposition for becoming vaccinated, and sources of information about COVID-19. A random sample, stratified by students, faculty, and administrative staff, was selected. The seroprevalence found was 2.64% (39/1479, 95% CI 1.8–3.4), and the adjusted seroprevalence was 2.89% (95% CI 2.1–3.7). The average age of the students was 23.2 years old, and 47.6 years old for staff. In relation to COVID-19, the following was found: 17.7% pauci-symptomatic, 1.3% symptomatic, 5.5% contact with cases, 4.9% confined, and 0.3% PCR positive. More than 90% complied with preventive measures. The proportion willing to receive the COVID-19 vaccine was 91%. Their sources of information were the Internet (74%) and television (70.1%). They requested that the university offer information (45.1%), training (27%), and provide Personal Protective Equipment (PPE) (26.3%). Lastly, 87.9% would repeat the test. A plan was established that included the follow-up of cases and contacts, random sample testing, training courses, bimodal teaching, a specific website, and the distribution of PPE.

Este artículo se encuentra disponible en la siguiente URL: https://www.mdpi.com/1660-4601/18/7/3356 Este artículo pertenece al número especial "Nursing research". Health professionals are the most influential and main sources of information about vaccines for the general population, as they are regarded as role models by patients and society. The objective of the present study was to determine the knowledge and attitudes of a group of university Nursing students about vaccines, as well as their sources of information and their education needs. A cross-sectional study was performed through a questionnaire (55 items) provided to Nursing students at two Spanish universities. A total of 1122 students participated in the study. The mean score obtained for knowledge about vaccines was 44.6 4.3, and for attitudes towards vaccines, it was 37.2 3.9. Hepatitis B (94.7%) and the Flu (89%) are the two main vaccines they should receive as health workers. The main source of information was the family environment (65.6%). Most of them considered that post-graduate education about vaccines should be provided by academic entities (universities, 62.7%). Among the health professionals, Nurses (85.5%) must be better educated and trained on the subject of vaccines. It is therefore necessary to delve into and complete the nurses’ training on vaccines, to educate them about the risks at the individual level, and their decisive role as promoters of the vaccination strategy for the general population. Universities must become the leaders in vaccine education and training.

Vaccine safety surveillance is essential in vaccination programs. We accomplished a descriptive study of surveillance AEFI-reporting rate in human papillomavirus (HPV) vaccine administered in the Valencian Community, Spain. Data were obtained from Spanish Pharmacovigilance Adverse Reactions Data (FEDRA). Reporting rates were calculated using local net doses distributed as the denominator. Trends were assessed using joinpoint regression with annual percent change (APC) reported. The AEFI-reports decreased between 2008 and 2018 in two periods, a fast decreasing rate from 2009 to 2011 (from 192.2 to 24.93 per 100000 doses, APC, &minus, 54.9%, 95%CI [&minus, 75.2, &minus, 17.7]), followed by a stable trend (&minus, 13% APC, 95%CI [&minus, 26.1, 2.4]). For the age group analysis, only the group aged 14&ndash, 15 years old followed the same trend with -58.4% (95%CI [&minus, 73.9, 33.8]) APC during 2008&ndash, 2011, and &minus, 8.8% (95%CI [&minus, 27.7, 15]) APC during 2011-2018. The majority of the reports (73.82%) were nonserious, involving reactions at or near the vaccination site, headache, and dizziness events. No death was reported. AEFI-reporting rates for HPV immunization in the Valencian Community have decreased considerably with two trend periods observed for girls aged 14&ndash, 15 years old. Currently, the AEFI reporting rate shows a decreasing trend, perhaps following the Weber effect, and it could also be affected by media attention and coverage.

The study aims to identify the relevance of immunohistochemistry (IHC), copy number aberrations (CNA) and epigenetic disorders in BRCAness breast cancers (BCs). We studied 95 paraffin included BCs, of which 41 carried BRCA1/BRCA2 germline mutations and 54 were non hereditary (BRCAX/Sporadic). Samples were assessed for BRCA1ness and CNAs by Multiplex Ligation-dependent Probe Amplification (MLPA); promoter methylation (PM) was assessed by methylation-specific-MLPA and the expression of miR-4417, miR-423-3p, miR-590-5p and miR-187-3p by quantitative RT-PCR. IHC markers Ki67, ER, PR, HER2, CK5/6, EGFR and CK18 were detected with specific primary antibodies (DAKO, Denmark). BRCAness association with covariates was performed using multivariate binary logistic regression (stepwise backwards Wald option). BRCA1/2 mutational status (p = 0.027), large tumor size (p = 0.041) and advanced histological grade (p = 0.017) among clinic-pathological variables; ER (p < 0.001) among IHC markers; MYC (p < 0.001) among CNA; APC (p = 0.065), ATM (p = 0.014) and RASSF1 (p = 0.044) among PM; and miR-590-5p (p = 0.001), miR-4417 (p = 0.019) and miR-423 (p = 0.013) among microRNA expression, were the selected parameters significantly related with the BRCAness status. The logistic regression performed with all these parameters selected ER+ as linked with the lack of BRCAness (p = 0.001) and MYC CNA, APC PM and miR-590-5p expression with BRCAness (p = 0.014, 0.045 and 0.007, respectively). In conclusion, the parameters ER expression, APC PM, MYC CNA and miR-590-5p expression, allowed detection of most BRCAness BCs. The identification of BRCAness can help establish a personalized medicine addressed to predict the response to specific treatments.

The correct immunization of the inmate population minimizes the risk of transmission of vaccine-preventable diseases in prisons. The objective of this study was to evaluate the vaccine coverage of long-term prisoners in the Spanish penitentiary system through a retrospective longitudinal study. One-thousand and five prisoners were selected, who were imprisoned from 2008 and 2018 in three Spanish prisons. Their degree of immunization was evaluated as related to hepatitis A (HAV), hepatitis B (HBV), tetanus, diphtheria, pneumococcus and seasonal flu. The state of vaccination of the prisoners with a serological diagnosis of HBV, hepatitis C (HCV) and human immunodeficiency virus (HIV) was also evaluated. The vaccination coverage obtained for hepatitis B was 52.3%, and for tetanus&ndash, diphtheria, it was 71.9%. However, for hepatitis A and pneumococcus infection, it was insignificant (&lt, 2% of the prisoners). Vaccination against seasonal flu was lower than 16%. The HCV and HIV-positive inmates were not correctly vaccinated either. The insufficient level of immunization obtained reflects the lack of interest and marginalization of this population by the penitentiary system and the health authorities. The lack of reliable records is combined with the lack of planned strategies that promote stable and well-defined programs of active vaccination.

Purpose: A recent study reported that 5-fluorouracil (5-FU)-based chemotherapy is less effective in treating patients with advanced colorectal cancer demonstrating hypermethylation of the TFAP2E gene. The aim of our study was to confirm and validate these findings in large, uniformly treated, well-characterized patient cohorts. Experimental Design: Two cohorts of 783 patients with colorectal cancer: 532 from a population-based, multicenter cohort (EPICOLON I) and 251 patients from a clinic-based trial were used to study the effectiveness of TFAP2E methylation and expression as a predictor of response of colorectal cancer patients to 5-FU–based chemotherapy. DNA methylation status of the TFAP2E gene in patients with colorectal cancer was assessed by quantitative bisulfite pyrosequencing analysis. IHC analysis of the TFAP2E protein expression was also performed. Results: Correlation between TFAP2E methylation status and IHC staining was performed in 607 colorectal cancer samples. Among 357 hypermethylated tumors, only 141 (39.6%) exhibited loss of protein expression. Survival was not affected by TFAP2E hypermethylation in stage IV patients [HR, 1.21; 95% confidence interval (CI), 0.79–1.87; log-rank P = 0.6]. In stage II–III cases, disease-free survival was not influenced by TFAP2E hypermethylation status in 5-FU–treated (HR, 0.91; 95% CI, 0.52–1.59; log-rank P = 0.9) as well as in nontreated patients (HR, 0.88; 95% CI, 0.5–1.54; log-rank P = 0.7). Conclusions: TFAP2E hypermethylation does not correlate with loss of its protein expression. Our large, systematic, and comprehensive study indicates that TFAP2E methylation and expression may not play a major role in predicting response to 5-FU–based chemotherapy in patients with colorectal cancer. Clin Cancer Res; 24(12); 2820–7. ©2018 AACR.

Objective The aim of this study was to validate a molecular classification of colorectal cancer (CRC) based on microsatellite instability (MSI), CpG island methylator phenotype (CIMP) status, BRAF, and KRAS and investigate each subtype's response to chemotherapy. Design This retrospective observational study included a population-based cohort of 878 CRC patients. We classified tumours into five different subtypes based on BRAF and KRAS mutation, CIMP status, and MSI. Patients with advanced stage II (T4N0M0) and stage III tumours received 5-fluoruracil (5-FU)-based chemotherapy or no adjuvant treatment based on clinical criteria. The main outcome was disease-free survival (DFS). Results Patients with the combination of microsatellite stable (MSS) tumours, BRAF mutation and CIMP positive exhibited the worst prognosis in univariate (log rank P < 0.0001) and multivariate analyses (hazard ratio 1.75, 95% CI 1.05-2.93, P = 0.03) after adjusting for age, sex, chemotherapy, and TNM stage. Treatment with 5-FU-based regimens improved prognosis in patients with the combination of MSS tumours, KRAS mutation and CIMP negative (log rank P = 0.003) as well as in patients with MSS tumours plus BRAF and KRAS wild-type and CIMP negative (log-rank P < 0.001). After adjusting for age, sex, and TNM stage in the multivariate analysis, only patients with the latter molecular combination had independently improved prognosis after adjuvant chemotherapy (hazard ratio 2.06, 95% CI 1.24-3.44, P = 0.005). Conclusion We confirmed the prognostic value of stratifying CRC according to molecular subtypes using MSI, CIMP status, and somatic KRAS and BRAF mutation. Patients with traditional chromosomally unstable tumours obtained the best benefit from adjuvant 5-FU-based chemotherapy. This work was supported by the Instituto de Salud Carlos III (PI08/0726, INT-09/208, PI11/2630, INT-12-078, INT13-196, PI14/01386), FISABIO-ISABIAL foundation (UGP-13-221, UGP-14-265), and the Asociación Española contra el Cáncer (Fundación Científica GCB13131592CAST). Eva Hernández-Illán received a grant from Instituto de Salud Carlos III (FI12/00233). Mar Giner-Calabuig received a grant from VALi+d. EXP ACIF/2016/002. Miren Alustiza received a predoctoral grant from ISABIAL (UGP-16-138). Other financial support was obtained from Asociación Española de Gastroenterología grants (Gonzalo Miño 2014, Tamarite 2015, Grupo de endoscopia 2016).

555 Background: The role of genetic profile of polyps to predict advanced metachronous lesions (AML) remains unknown. The aim is to study the relation between genetic profile of polyps and both risk of AMLs and time to develop them in surveillance. Methods: 308 patients with colonic polyps were consecutively enrolled between 2007 and 2009 for this cohort study, and followed up to 2014 (median 26 months, range 63). Variables as age, sex, smoking, weight, number of colonoscopies and number and characteristics of polyps were collected. 995 polyps were analyzed for somatic mutations on BRAF and KRAS genes using allelic discrimination by real-time PCR and direct DNA sequenciation, respectively. High level of methylation on CpG islands (CIMP-H) was also tested using MS-MLPA. AML was defined by a size higher than 9mm, high grade dysplasia or villous component. Risk of developing AML for individual genetic markers was studied using Chi-square tests and logistic regression. Log-rank test with Kaplan Meier survival curves and Cox-regression model were also performed. Multivariate analysis were adjusted by sex, age, familial colorectal cancer, smoking and features of AML in first colonoscopy. Results: 21% of polyps in first colonoscopy were CIMP-H. KRAS and BRAF mutations accounted for 25% and 17% of polyps, respectively. In univariate analysis, KRAS-mutated polyps were related to higher risk of AML in surveillance (52% KRAS-mutated polyps vs 31% non-mutated; p = 0.01). Similar results were obtained regarding CIMP-H (77% CIMP-H polyps vs 38% non-CIMP; p = 0.005). Logistic regression showed CIMP-H as the unique genetic marker of risk for AML (OR 11.41, 95% CI 2.04-63.70; p = 0.006). Regarding time to develop AML, shorter intervals were found related to CIMP-H (median of 31 vs 48 months in non-CIMP-H; p = 0.002) and KRAS-mutations (median of 36 vs 49 months in non-mutated; p = 0.029) in univariate analysis. Multivariate analysis highlighted CIMP-H as the unique independent marker associated to shorter time to develop AML (HR 4.01, 95% CI 1.36-10.46; p = 0.01). Conclusions: Presence of CIMP-H in polyps associates higher risk of subsequent AML and shorter interval to their development. Genetic profile of polyps emerges as useful tool for colonoscopy surveillance.

Purpose: The present study aimed to determine the prevalence of MUTYH mutations in patients with multiple colonic polyps and to explore the best strategy for diagnosing MUTYH-associated polyposis (MAP) in these patients. Experimental Design: This study included 405 patients with at least 10 colonic polyps each. All cases were genetically tested for the three most frequent MUTYH mutations. Whole-gene analysis was performed in heterozygous patients and in 216 patients lacking the three most frequent mutations. Polyps from 56 patients were analyzed for the KRAS-Gly12Cys and BRAF V600E somatic mutations. Results: Twenty-seven (6.7%) patients were diagnosed with MAP, of which 40.8% showed serrated polyps. The sensitivity of studying only the three common variants was 74.1%. Of 216 patients without any monoallelic mutation in common variants, whole-gene analysis revealed biallelic pathogenic mutation in only one. G396D mutation was associated with serrated lesions and older age at diagnosis. There was a strong association between germinal MUTYH mutation and KRAS Gly12Cys somatic mutation in polyps. BRAF V600E mutation was found in 74% of serrated polyps in MUTYH-negative patients and in none of the polyps of MAP patients. Conclusions: We observed a low frequency of MUTYH mutations among patients with multiple adenomatous and serrated polyps. The MAP phenotype frequently included patients with serrated polyps, especially when G396D mutation was involved. Our results show that somatic molecular markers of polyps can be useful in identifying MAP cases and support the need for the complete MUTYH gene analysis only in patients heterozygous for recurrent variants. Clin Cancer Res; 20(5); 1158–68. ©2014 AACR.

This study aims to identify signatures of miR associated with hereditary, BRCA1 or BRCA2 mutation positive breast cancer (BC), and non-hereditary BC, either sporadic (SBC) or non-informative (BRCAX). Moreover, we search for signatures associated with tumor stage, immunohistochemistry and tumor molecular profile. Twenty formalin fixed paraffin embedded (FFPE) BCs, BRCA1, BRCA2, BRCAX and SBC, five per group were studied. Affymetrix platform miRNA v.3.0 was used to perform miR expression analysis. ER, PR, HER2 and Ki67 protein expression was analyzed by immunohistochemistry. BRCA1, BRCA2 and RASSF1 methylation analysis, AURKA copy number variations, and BRCA1 and BRCA2 deletions, were studied by MLPA. We validated eight of the miR selected by the arrays in 77 BCs by qRT-PCR. The miR profiles associated with tumor features were studied applying the Sparse Partial Least Squares Discriminant Analysis. MiR discrimination capability to distinguish hereditary and non-hereditary BC was analyzed by the discriminant function. With 15 out of 1,733 hsa-miRs, it was possible to differentiate the four groups. BRCA1, BRCA2 and SBC were associated with clusters of hyper-expressed miRs, and BRCAX with hypo-expressed miRs. Hsa-miR-4417 and hsa-miR-423-3p expressions (included among the eight validated miRs) differentiated 70.1 % of hereditary and non-hereditary BCs. We found miR profiles associated with tumor features like node involvement, histological grade, ER, PR and HER2 expression. Regarding molecular parameters, we only found a weak association of miRs in BC harboring losses in AURKA. We conclude that array miR expression profiles can differentiate the four study groups using FFPE BC. However, miRs expression estimated by qRT-PCR differentiates only hereditary and non-inherited BCs. The miR expression array is a simple and rapid approach that could be useful to facilitate the identification of those SBC carrying genetic or epigenetic changes in BRCA genes responsible of BRCA-like phenotype. These patients could benefit from the treatment with PARP inhibitors.

Background & Aims We investigated whether patients with multiple serrated polyps, but not meeting the World Health Organization criteria for serrated polyposis syndrome, and their relatives have similar risks for colorectal cancer (CRC) as those diagnosed with serrated polyposis. Methods We collected data from patients with more than 10 colonic polyps, recruited in 2008–2009 from 24 hospitals in Spain for a study of causes of multiple colonic polyps. We analyzed data from 53 patients who met the criteria for serrated polyposis and 145 patients who did not meet these criteria, but who had more than 10 polyps throughout the colon, of which more than 50% were serrated. We calculated age- and sex-adjusted standardized incidence ratios (SIRs) for CRC in both groups, as well as in their first-degree relatives. Results The prevalence of CRC was similar between patients with confirmed serrated polyposis and multiple serrated polyps (odds ratio, 1.35; 95% confidence interval [CI], 0.64–2.82; P = .40). The SIR for CRC in patients with serrated polyposis (0.51; 95% CI, 0.01–2.82) did not differ significantly from the SIR for CRC in patients with multiple serrated polyps (0.74; 95% CI, 0.20–1.90; P = .70). The SIR for CRC also did not differ significantly between first-degree relatives of these groups (serrated polyposis: 3.28, 95% CI, 2.16–4.77; multiple serrated polyps: 2.79, 95% CI, 2.10–3.63; P = .50). Kaplan–Meier analysis showed no differences in the incidence of CRC between groups during the follow-up period (log-rank, 0.6). Conclusions The risk of CRC in patients with multiple serrated polyps who do not meet the criteria for serrated polyposis, and in their first-degree relatives, is similar to that of patients diagnosed with serrated polyposis.

Background and study aims: Endoscopic surveillance in patients with multiple colorectal polyps aims to reduce colorectal cancer (CRC) incidence and mortality, as well as the need for colorectal surgery. The aim of this study was to determine the risk of developing CRC or the need for surgery during endoscopic surveillance in a cohort of patients with multiple (10-100) colorectal polyps. Patients and methods: This was a multicentrer, longitudinal, observational study in 15 CRC high risk clinics in Spain, carried out between January 2009 and December 2010. Patients who were included in the EPIPOLIP trial and had at least 1 year of follow-up were included in the study. The primary outcome of interest was the incidence of CRC at least 1 year following the initial colonoscopy. The secondary outcome was the need for colorectal surgery. Results: A total of 265 patients were followed for a median of 3.8 years. Patients underwent a median of 5 colonoscopies, and 17 patients (6.4 %) were diagnosed with CRC. A total of 32 patients (12.1 %) underwent surgery, including 15 (5.7 %) for prophylaxis without a diagnosis of CRC. The corresponding incidence density rates for CRC and colorectal surgery were 1.4 (95% confidence interval [CI] 0.7 to 2.1) and 2.7 (95 % CI 1.7 to 3.6) per 100 patient-years, respectively. Only the presence of symptoms at first colonoscopy was independently associated with CRC diagnosis (hazard ratio [HR] 7.7, 95% CI 1.1 to 59.3) and colorectal surgery (HR 4.6, 95% CI 1.02 to 20.6). Conclusions: Patients with more than 10 neoplastic polyps required frequent colonoscopies within a short follow-up period. More than 10% of patients required colorectal surgery within 4 years, more than half for incident CRC.

Molecular advances support the existence of an alternative pathway of colorectal carcinogenesis that is based on the hypermethylation of specific DNA regions that silences tumor suppressor genes. This alternative pathway has been called the serrated pathway due to the serrated appearance of tumors in histological analysis. New classifications for colorectal cancer (CRC) were proposed recently based on genetic profiles that show four types of molecular alterations: BRAF gene mutations, KRAS gene mutations, microsatellite instability, and hypermethylation of CpG islands. This review summarizes what is known about the serrated pathway of CRC, including CRC molecular and clinical features, prognosis, and response to chemotherapy.

Background: The selection of patients for genetic testing to rule out Lynch syndrome is currently based on fulfilment of at least one of the revised Bethesda criteria followed by mismatch repair (MMR) status analysis. A study was undertaken to compare the present approach with universal MMR study-based strategies to detect Lynch syndrome in a large series of patients with colorectal cancer (CRC). Methods: 2093 patients with CRC from the EPICOLON I and II cohorts were included. Immunohistochemistry for MMR proteins and/or microsatellite instability (MSI) analysis was performed in tumour tissue. Germline MLH1 and MSH2 mutation analysis was performed in patients whose tumours showed loss of MLH1 or MSH2 staining, respectively. MSH6 genetic testing was done in patients whose tumours showed lack of MSH6 expression or a combined lack of MSH2 and MSH6 expression but did not have MSH2 mutations. PMS2 genetic testing was performed in patients showing isolated loss of PMS2 expression. In patients with MSI tumours and normal or not available MMR protein expression, all four MMR genes were studied. Results: A total of 180 patients (8.6%) showed loss of expression of some of the MMR proteins and/or MSI. Four hundred and eighty-six patients (23.2%) met some of the revised Bethesda criteria. Of the 14 (0.7%) patients who had a MMR gene mutation, 12 fulfilled at least one of the revised Bethesda criteria and two (14.3%) did not. Conclusions: Routine molecular screening of patients with CRC for Lynch syndrome using immunohistochemistry or MSI has better sensitivity for detecting mutation carriers than the Bethesda guidelines. Instituto de Salud Carlos III (INT09/208 and PI08/0726). Fundación de la CV para la Investigación en el Hospital General Universitario de Alicante. Grant SAF 07-64873 from the Ministerio de Educación y Ciencia. Funds from the AGAUR (2009 SGR 849) Instituto de Salud Carlos III (FI07/00303) CIBERehd is funded by the Instituto de Salud Carlos III. Grant from Conselleria d’Educació de la Generalitat Valenciana (VALi+d).

Resumen de la comunicacion presentada en 4th Biennial Meeting : International Society for Gastrointestinal Hereditary Tumours San Antonio, Texas, March 31-April 2, 2011.

This study aims to identify the profile of immunohistochemical (IHC) parameters, copy number aberrations (CNAs) and epigenetic alterations [promoter methylation (PM) and miR expression] related to hereditary (H) and triple negative (TN) breast cancer (BC). This profile could be of relevance for guiding tumor response to treatment with targeting therapy. The study comprises 278 formalin fixed paraffin-embedded BCs divided into two groups: H group, including 88 hereditary BC (HBC) and 190 non hereditary (NHBC), and TN group, containing 79 TNBC and 187 non TNBC (NTNBC). We assessed IHC parameters (Ki67, ER, PR, HER2, CK5/6, CK18 and Cadherin-E), CNA of 20 BC related genes, and PM of 24 tumor suppressor genes employing MLPA/MS-MLPA (MRC Holland, Amsterdam). MiR-4417, miR-423-3p, miR-590-5p and miR-187-3p expression was assessed by quantitative RT-PCR (Applied Biosystems). Binary logistic regression was applied to select the parameters that better differentiate the HBC or TN groups. For HBC we found that, ER expression, ERBB2 CNA and PM in RASSF1 and TIMP3 were associated with NHBC whereas; MYC and AURKA CNA were linked to HBC. For TNBC, we found that CDC6 CNA, GSTP1 and RASSF1 PM and miR-423-3p hyperexpression were characteristic of NTNBC, while MYC aberrations, BRCA1 hypermethylation and miR-590-5p and miR-4417 hyperexpression were more indicative of TNBC. The selected markers allow establishing BC subtypes, which are characterized by showing similar etiopathogenetic mechanisms, some of them being molecular targets for known drugs or possible molecular targets. These results could be the basis to implement a personalized therapy.

Background The prevalence of MLH1 constitutional epimutations in the general population is unknown. We sought to analyse the prevalence of MLH1 constitutional epimutations in unselected and selected series of patients with colorectal cancer (CRC). Methods Patients with diagnoses of CRC (n=2123) were included in the unselected group. For comparison, a group of 847 selected patients with CRC who fulfilled the revised Bethesda guidelines (rBG) were also included. Somatic and constitutional MLH1 methylation was assayed via methylation-specific multiplex ligation-dependent probe amplification of cases lacking MLH1 expression. Germline alterations in mismatch-repair (MMR) genes were assessed via Sanger sequencing and methylation-specific multiplex ligation-dependent probe amplification. Results Loss of MLH1 expression occurred in 5.5% of the unselected series and 12.5% of the selected series (p

Background & aims High-risk features of colonic polyps are based on size, number, and pathologic characteristics. Surveillance colonoscopy is often recommended according to these findings. This study aimed to determine whether the molecular characteristics of polyps might provide information about the risk of metachronous advanced neoplasia. Methodology We retrospectively included 308 patients with colonic polyps. A total of 995 polyps were collected and tested for somatic BRAF and KRAS mutations. Patients were classified into 3 subgroups, based on the polyp mutational profile at baseline, as follows: non-mutated polyps (Wild-type), at least one BRAF-mutated polyp, or at least one KRAS-mutated polyp. At surveillance, advanced adenomas were defined as adenomas >= 10 mm and/or with high grade dysplasia or a villous component. In contrast, advanced serrated polyps were defined as serrated polyps >= 10 mm in any location, located proximal to the splenic flexure with any size or with dysplasia. Results At baseline, 289 patients could be classified as wild-type (62.3%), BRAF mutated (14.9%), or KRAS mutated (22.8%). In the univariate analysis, KRAS mutations were associated with the development of metachronous advanced polyps (OR: 2.36, 95% CI: 1.22-4.58; P= 0.011), and specifically, advanced adenomas (OR: 2.42, 95% CI: 1.13-5.21; P= 0.023). The multivariate analysis, adjusted for age and sex, also showed associations with the development of metachronous advanced polyps (OR: 2.27, 95% CI: 1.15-4.46) and advanced adenomas (OR: 2.23, 95% CI: 1.02-4.85). Conclusions Our results suggested that somatic KRAS mutations in polyps represent a potential molecular marker for the risk of developing advanced neoplasia.

Germline mutations in DNA polymerase E > (POLE) and delta (POLD1) have been recently identified in families with multiple colorectal adenomas and colorectal cancer (CRC). All reported cases carried POLE c.1270C > G (p.Leu424Val) or POLD1 c.1433G > A (p.Ser478Asn) mutations. Due to the scarcity of cases reported so far, an accurate clinical phenotype has not been defined. We aimed to assess the prevalence of these recurrent mutations in unexplained familial and early-onset CRC and polyposis, and to add additional information to define the clinical characteristics of mutated cases. A total of 858 familial/early onset CRC and polyposis patients were studied: 581 familial and early-onset CRC cases without mismatch repair (MMR) deficiency, 86 cases with MMR deficiency and 191 polyposis cases. Mutation screening was performed by KASPar genotyping assays and/or Sanger sequencing of the involved exons. POLE p.L424V was identified in a 28-year-old polyposis and CRC patient, as a de novo mutation. None of the 858 cases studied carried POLD1 p.S478N. A new mutation, POLD1 c.1421T > C (p.Leu474Pro), was identified in a mismatch repair proficient Amsterdam II family. Its pathogenicity was supported by cosegregation in the family, in silico predictions, and previously published yeast assays. POLE and POLD1 mutations explain a fraction of familial CRC and polyposis. Sequencing the proofreading domains of POLE and POLD1 should be considered in routine genetic diagnostics. Until additional evidence is gathered, POLE and POLD1 genetic testing should not be restricted to polyposis cases, and the presence of de novo mutations, considered.

Background: Lynch syndrome (LS) is a hereditary condition that increases the risk for endometrial and other cancers. The identification of endometrial cancer (EC) patients with LS has the potential to influence life-saving interventions. We aimed to study the prevalence of LS among EC patients in our population. Methods: Universal screening for LS was applied for a consecutive series EC. Tumor testing using microsatellite instability (MSI), immunohistochemistry (IHC) for mismatch-repair (MMR) protein expression and MLH1-methylation analysis, when required, was used to select LS-suspicious cases. Sequencing of corresponding MMR genes was performed. Results: One hundred and seventy-three EC (average age, 63 years) were screened. Sixty-one patients (35%) had abnormal IHC or MSI results. After MLH1 methylation analysis, 27 cases were considered suspicious of LS. From these, 22 were contacted and referred for genetic counseling. Nineteen pursued genetic testing and eight were diagnosed of LS. Mutations were more frequent in younger patients (

Background & Aims We investigated clinical and molecular differences between the different phenotypes of serrated polyposis syndrome (SPS) and the frequency of mutations in BRAF or KRAS in polyps from patients with SPS. Methods We collected data on clinical and demographic characteristics of 50 patients who fulfilled the criteria for SPS. Polymerase chain reaction and sequence analysis were used to identify BRAF and KRAS mutations in 432 polyps collected from 37 patients; we analyzed CpG island methylator phenotypes in 272 of these polyps. Results Fifteen patients (30%) had type 1 SPS and 35 had type 2 SPS. There were no significant differences in age at diagnosis, sex, smoking frequency, body mass index, or colorectal cancer predisposition between groups of patients, or in the pathologic or molecular characteristics of their polyps. A familial history of colorectal cancer or colonic polyps was reported more frequently by patients with type 2 SPS. BRAF mutations were found in 63% of polyps and KRAS mutations were found in 9.9%; 43.4% of polyps had the CpG island methylator phenotype–high phenotype. A per-patient analysis revealed that all patients had a BRAF or KRAS mutation in more than 25% of their polyps; 84.8% of patients had a mutation in BRAF or KRAS in more than 50% of their polyps. Conclusions Except for a greater likelihood of familial history of colorectal cancer or colonic polyps in patients with type 2 SPS, we found no significant demographic, pathologic, or molecular differences between types 1 and 2 SPS. All patients had a BRAF or KRAS mutation in at least 25% of their polyps.

In colorectal cancer (CRC), an inherited susceptibility risk affects about 35% of patients, whereas high-penetrance germline mutations account for

Background Lynch syndrome (LS) is an autosomal dominant inherited cancer syndrome characterized by early onset cancers of the colorectum, endometrium and other tumours. A significant proportion of DNA variants in LS patients are unclassified. Reports on the pathogenicity of the c.1852_1853AA>GC (p.Lys618Ala) variant of the MLH1 gene are conflicting. In this study, we provide new evidence indicating that this variant has no significant implications for LS. Methods The following approach was used to assess the clinical significance of the p.Lys618Ala variant: frequency in a control population, case-control comparison, co-occurrence of the p.Lys618Ala variant with a pathogenic mutation, co-segregation with the disease and microsatellite instability in tumours from carriers of the variant. We genotyped p.Lys618Ala in 1034 individuals (373 sporadic colorectal cancer [CRC] patients, 250 index subjects from families suspected of having LS [revised Bethesda guidelines] and 411 controls). Three well-characterized LS families that fulfilled the Amsterdam II Criteria and consisted of members with the p.Lys618Ala variant were included to assess co-occurrence and co-segregation. A subset of colorectal tumour DNA samples from 17 patients carrying the p.Lys618Ala variant was screened for microsatellite instability using five mononucleotide markers. Results Twenty-seven individuals were heterozygous for the p.Lys618Ala variant; nine had sporadic CRC (2.41%), seven were suspected of having hereditary CRC (2.8%) and 11 were controls (2.68%). There were no significant associations in the case-control and case-case studies. The p.Lys618Ala variant was co-existent with pathogenic mutations in two unrelated LS families. In one family, the allele distribution of the pathogenic and unclassified variant was in trans, in the other family the pathogenic variant was detected in the MSH6 gene and only the deleterious variant co-segregated with the disease in both families. Only two positive cases of microsatellite instability (2/17, 11.8%) were detected in tumours from p.Lys618Ala carriers, indicating that this variant does not play a role in functional inactivation of MLH1 in CRC patients. Conclusions The p.Lys618Ala variant should be considered a neutral variant for LS. These findings have implications for the clinical management of CRC probands and their relatives. This action has been supported in part by grants from the Generalitat Valenciana in Spain (AP140/08) and the Biomedical Research Foundation from the Hospital of Elche, Spain (FIBElx0902). CG, AMC, CEl and LPC are recipients of fellowships from the Conselleria de Educació (Generalitat Valenciana); Fundacion Juan Peran-Pikolinos; Fundacion Carolina-BBVA and Fondo Investigación Sanitaria (FI07/00303), respectively. RJ is receptor of a grant from Instituto de Salud Carlos III (INT09/208) SI

Summary Background The incidence of colorectal cancer (CRC) in Peru has been increasing, and no data have been published on the molecular features. We explored the most relevant genetic events involved in colorectal carcinogenesis, with clinical implications. Methods Using immunohistochemistry for mismatch-repair (MMR) proteins (MLH1, MSH2, MSH6, and PMS2) and microsatellite instability analysis, we evaluated the status of 90 non-selected CRC Peruvian patients followed in a nationwide reference hospital for cancer (INEN, Lima). Tumours with loss of hMLH1 were evaluated further for hMLH1 promoter hypermethylation and all cases were evaluated for the presence of KRAS and BRAF-V600E mutations. Results MMR deficiency was found in 35 (38.8%) patients. We identified an unexpected association between MMR deficiency and older age. Among the 14 cases with loss of MLH1, 10 samples exhibited hypermethylation. Of the 90 cases evaluated, 15 (16.7%) carried KRAS mutations; we found one previously unreported mutation (G13R). Conclusions Peruvian CRC tumours exhibited the highest prevalence of MMR deficiency reported to date. The expected hereditary component was also high. The age of onset of these MMR deficient tumours was greater than that observed for non-MMR deficient cases, suggesting the ineffectiveness of the Bethesda criteria for Lynch syndrome screening in Peru. Prospective studies are warranted to define the molecular characteristics of CRC in this population.

The standard genetic test for Lynch syndrome (LS) frequently reveals an absence of pathogenic mutations in DNA mismatch repair genes known to be associated with LS. It was recently shown that germ line deletions in the last exons of EPCAM are involved in the etiology of LS. The aim of this study was to evaluate the prevalence of EPCAM deletions in a Spanish population and the clinical implications of deletion. Probands from 501 families suspected of having LS were enrolled in the study. Twenty-five cases with MSH2 loss were identified: 10 had mutations of MSH2, five had mutations of MSH6, and 10 did not show MSH2/MSH6 mutations. These 25 cases were analyzed for EPCAM deletions using multiplex ligation-dependent probe amplification, and deletions were mapped using long-range PCR analysis. One subject with no MSH2/MSH6 mutations had a large deletion in the EPCAM locus that extended for 8.7 kb and included exons 8 and 9. The tumor exhibited MSH2 promoter hypermethylation. EPCAM deletion analysis followed by MSH2 methylation testing of the tumor is a fast low-cost procedure that can be used to identify mutations that cause LS. We propose that this procedure be incorporated into clinical genetic analysis strategies and present a decision-support flow diagram for the diagnosis of LS. Transversal Cancer Action (ISCIII) and the Biomedical Research Foundation from the Elche University Hospital. Carolina-BBVA Foundation and Juan Peran-Pikolinos Foundation.

in 32 (15.4%) AJ and 22 (0.5%) NAJ (p=0.0001); among AJ this variant was significantly associated with a diagnosis of CRC (OR: 2.6 [95% CI: 1.1-5.8]). Conclusion: Overall, the frequency of pathogenic APC or MUTYH mutations increases with adenoma count and is lower in AJ compared to NAJ. The low yield of comprehensive MUTYH testing in AJ suggests this gene plays a small role in AJ with polyposis and that other genes may be responsible for the polyposis phenotype in Ashkenazim. The higher prevalence of the APC*R332X mutation in AJ compared to NAJ suggests R332X may be a founder mutation. This should be further explored through haplotype analysis. Finally, we propose that increased colorectal screening be considered in Ashkenazim who have the I1307K variant and report a personal history of colorectal adenomas.

BACKGROUND. Early and non-invasive detection of gastric cancer (GC) is remaining a challenge in many high-risk areas. OBJECTIVES. To investigate the feasibility of gastric cancer and peptic ulcer disease detection by measuring volatile organic compounds (VOCs) in the exhaled breath by a nanomaterial-based sensor technology. METHODS. Alveolar exhaled breath samples were collected from 99 GC patients, 53 peptic ulcer disease (PUD) patients and 342 controls in Latvia (Caucasian population) having been investigated by upper endoscopy. Nanomaterial-based sensors Gold nano particles (GNP) and single-wall carbon nanotube (SWCNT) were used to discriminate the GC and the PUD groups from the healthy controls by loading discriminant factor analysis (DFA) pattern recognition. Classification success was calculated by (i) building an algorithm for 70% of the samples as a training set and (ii) randomly blinding 30% of the samples as a validation set. RESULTS. The blind DFA models showed: (i) An excellent discrimination between the GC patients and controls (91% accuracy); (ii) An excellent discrimination between the GC patients and PUD patients (86% accuracy); (iii) Good discrimination between the PUD patients and controls (80% accuracy). CONCLUSIONS: The obtained results are demonstrating the good potential of VOC detection in exhaled breath by nanomaterial-based sensor technology in diagnosing GC and PUD. Performance characteristics of nanomaterial-based sensor technology to detect gastric cancer and peptic ulcer disease

at least one new peak compared to the product from the matching normal tissue. Statistical Analysis: The Kaplan-Meier method was used for estimating recurrence-free survival. Cox proportional hazard analysis was used to evaluate the association between MSI status and other clinicopathological factors for predicting recurrent distant metastasis. The P value less than 0.05 was considered to be statistically significant. Results: Stage II and III patients with MSI-L and/or EMAST had a shorter recurrence-free survival than patients with high levels of MSI (MSI-H) (P=0.0084) or with highly stable microsatellites (H-MSS) (P=0.0415) by Kaplan-Meier analysis. MSI-L and/or EMAST are independent predictors of recurrent distant metastasis in primary stage II and III CRCs by Cox proportional hazard analysis (Hazard Ratio: 1.83, 95%CI: 1.06-3.15, P=0.0301). Compatible with other studies, the present study also showed that MSI-H is associated with proximal location and exhibited lowest risk for recurrent distant metastasis. Conclusions: Our results showed that MSI-L/EMAST is a predictive factor of stage II/III primary CRC for recurrent distant metastasis. Because MSI-L/EMAST CRC is different from MSI-H and H-MSS CRC, we proposed to define MSI-L/EMAST as one group and named this group of CRC moderate MSI (MSI-M).

e14043 Background: CRC is the fifth and the fourth cause of cancer in Peruvian males and females, respectively. DNA mismatch repair (MMR) deficiency in CRC has been reported in a minority of cases ...

Diffuse large B-cell lymphoma (DLCL) exhibits heterogeneous clinical features and varies in response to treatment and prognosis. Establishment of parameters that can predict outcome could help to identify patients who may benefit from risk-adjusted therapies. Because apoptosis-related proteins may play an important role in predicting the prognosis of DLCL, the current study investigated the significance of bcl-2 expression in relation to clinical characteristics in this patients. We studied 125 patients (74/51 men/women; median age 64.17 years) consecutively diagnosed with de novo DLBCL in a single institution from Peru (HNGAI) during 3-year period (2000–2002).Morphology, and clinical characteristics were analyzed according to the primary site of the lymphoma and edge. Paraffin-embedded specimens from 88 were analyzed immunohistochemically for bcl-2 protein expression. Cases with a positive immunohistological stain in more than 20% of the tumor cells were considered positive expression. Results: Sites of the disease were: lymph node, 78 cases (62.4%); Waldeyer’s ring, 18 (14.4%); and extranodal sites, 45 (36%), including GI tract in 33 cases. 5-year overall survival (OS) was 19.7%. Log-rank analyses of survival showed significant differences between ≥60 years old and younger patients (P=0.003).bcl-2 expression was identified in 34 patients (38.6%), There was no significant difference in the OS (P=0.453) between the bcl-2+ (n=34) and bcl-2− (n=54) groups. In conclusion, bcl-2 expression appeared to be non predictive of a good OS in patients with DLCL, however, there was still a clear trend that the bcl-2+ patients died sooner than the patients with tumors that contained bcl-2− lymphoma cells. The current study show all patients as were staged or confined in a group with high or high intermediate IPI scores and could be candidates for alternative therapeutic approaches.