[COVID-19 vaccination at the University of Alicante (Spain): detection of neutralizing antibodies against SARS-CoV-2 by rapid serological test, a cross-sectional study]. / Vacunación contra la COVID-19 en la Universidad de Alicante (España): detección de anticuerpos neutralizantes frente al sars-CoV-2 mediante test serológico rápido, un estudio transversal.

OBJECTIVE: Neutralizing antibodies against SARS-CoV-2 have shown to be an effective tool for the analysis of the immunity generated against COVID-19. Numerous seroprevalence studies carried out in different groups have made it possible to draw a global map of vaccination coverage through the use of rapid lateral flow immunochromatography serological tests for clinical and epidemiological purposes. The objective of our study was to determine the degree of immunity against SARS-CoV-2 associated with the presence of neutralizing antibodies in administrative staff, teachers and students at the University of Alicante by means of a rapid serological test and to learn about their experience with vaccination against COVID-19. METHODS: A cross-sectional epidemiological study was designed, based on the prevalence of antibodies against the S protein (spike) of SARS-CoV-2. A total of 888 people participated. The study was carried out with a single test (July 6 to July 22, 2021). Using logistic regression, adjusted Odds Ratios were calculated according to sex, age, type of vaccine, number of vaccine doses received, complete vaccination schedule, and having had COVID-19. RESULTS: The vaccines received mostly were Vaxzevria® and Comirnaty®, with 73.3% between both, although 67.2% presented a complete regimen. The results of the OJABIO rapid neutralizing antibody test gave a positive result in 61.4% of the sample. There was a high association between the variables COVID-19 infection, two doses of vaccine, Spikevax® or Comirnaty® vaccine, and eighteen/twenty-nine years old group with a positive result on the OJABIO test. A total of 712 subjects answered the parallel survey (80%) on adverse effects and preferences between the different vaccines against COVID-19. CONCLUSIONS: The vaccination status against COVID-19 in the university community after six months of the start of national immunization strategies reflects low coverage despite the excellent willingness to get vaccinated. Neutralizing antibodies (NAb) rapid tests can be useful to guide immunization strategies and decide when to administer new booster doses.

OBJECTIVE: Los anticuerpos neutralizantes frente al SARS-CoV-2 han resultado una herramienta eficaz para el análisis de la inmunidad generada frente a la COVID-19. Numerosos estudios de seroprevalencia realizados en diferentes colectivos han permitido trazar un mapa global sobre la cobertura vacunal mediante el uso de pruebas serológicas rápidas de inmunocromatografía de flujo lateral con fines clínicos y epidemiológicos. El objetivo de nuestro estudio fue determinar el grado de inmunidad frente al SARS-CoV-2 asociado a la presencia de anticuerpos neutralizantes en personal administrativo, docentes y estudiantes de la Universidad de Alicante, mediante un test serológico rápido, así como conocer su experiencia sobre la vacunación frente a la COVID-19. METHODS: Se diseñó un estudio epidemiológico, transversal, basado en la prevalencia de anticuerpos frente a la proteína S (espícula o Spike) del SARS-CoV-2. Participaron un total de 888 personas. El estudio se llevó a cabo con un único test (6 de julio a 22 de julio de 2021). Mediante regresión logística se calcularon Odds Ratios ajustadas según sexo, edad, tipo de vacuna, número de dosis de vacuna recibidas, pauta completa de vacunación y haber padecido la COVID-19. RESULTS: Las vacunas recibidas mayoritariamente fueron Vaxzevria® y Comirnaty®, con un 73,3% entre ambas; el 67,2% presentó pauta completa. Los resultados del test rápido de anticuerpos neutralizantes OJABIO dieron un resultado positivo en el 61,4% de la muestra. La posibilidad de un resultado positivo en el test OJABIO estuvo fuertemente asociada a haber padecido la COVID-19, haber recibido dos dosis, estar vacunado con Spikevax® o Comirnaty® o pertenecer al grupo de dieciocho a veintinueve años. Un total de 712 sujetos respondieron a un cuestionario (80%) paralelo sobre los efectos adversos y las preferencias entre las distintas vacunas contra la COVID-19. CONCLUSIONS: El estado de vacunación frente a la COVID-19 en la comunidad universitaria a los seis meses de la puesta en marcha de las estrategias nacionales de inmunización refleja una baja cobertura asociada, a pesar de la excelente predisposición a vacunarse. Los test rápidos de anticuerpos neutralizantes (AcN) pueden ser de utilidad para orientar las estrategias de inmunización y para decidir el momento de administrar nuevas dosis de refuerzo.

Vacunación contra la COVID-19 en la Universidad de Alicante (España): detección de anticuerpos neutralizantes frente al SARS-CoV-2 mediante test serológico rápido, un estudio transversal / COVID-19 vaccination at the University of Alicante (Spain): detection of neutralizing antibodies against SARS-CoV-2 by rapid serological test, a cross-sectional study

Fundamentos: Los anticuerpos neutralizantes frente al SARS-CoV-2 han resultado una herramienta eficaz para el análisis de la inmunidad generada frente a la COVID-19. Numerosos estudios de seroprevalencia realizados en diferentes colectivos han permitido trazar un mapa global sobre la cobertura vacunal mediante el uso de pruebas serológicas rápidas de inmunocromatografía de flujo lateral con fines clínicos y epidemiológicos. El objetivo de nuestro estudio fue determinar el grado de inmunidad frente al SARS-CoV-2 asociado a la presencia de anticuerpos neutralizantes en personal administrativo, docentes y estudiantes de la Universidad de Alicante, mediante un test serológico rápido, así como conocer su experiencia sobre la vacunación frente a la COVID-19. Métodos: Se diseñó un estudio epidemiológico, transversal, basado en la prevalencia de anticuerpos frente a la proteína S (espícula o Spike) del SARS-CoV-2. Participaron un total de 888 personas. El estudio se llevó a cabo con un único test (6 de julio a 22 de julio de 2021). Mediante regresión logística se calcularon Odds Ratios ajustadas según sexo, edad, tipo de vacuna, número de dosis de vacuna recibidas, pauta completa de vacunación y haber padecido la COVID-19. Resultados: Las vacunas recibidas mayoritariamente fueron Vaxzevria® y Comirnaty ® , con un 73,3% entre ambas; el 67,2% presentó pauta completa. Los resultados del test rápido de anticuerpos neutralizantes OJABIO dieron un resultado positivo en el 61,4% de la muestra. La posibilidad de un resultado positivo en el test OJABIO estuvo fuertemente asociada a haber padecido la COVID-19, haber recibido dos dosis, estar vacunado con Spikevax® o Comirnaty® o pertenecer al grupo de dieciocho a veintinueve años. Un total de 712 sujetos respondieron a un cuestionario (80%) paralelo sobre los efectos adversos y las preferencias entre las distintas vacunas contra la COVID-19...(AU)

Background: Neutralizing antibodies against SARS-CoV-2 have shown to be an effective tool for the analysis of the immunity generated against COVID-19. Numerous seroprevalence studies carried out in different groups have made it possible to draw a global map of vaccination coverage through the use of rapid lateral flow immunochromatography serological tests for clinical and epidemiological purposes. The objective of our study was to determine the degree of immunity against SARS-CoV-2 associated with the presence of neutralizing antibodies in administrative staff, teachers and students at the University of Alicante by means of a rapid serological test and to learn about their experience with vaccination against COVID-19. Methods: A cross-sectional epidemiological study was designed, based on the prevalence of antibodies against the S protein (spike) of SARS-CoV-2. A totalof 888 people participated. The study was carried out with a single test (July 6 to July 22, 2021). Using logistic regression, adjusted Odds Ratios were calculated according to sex, age, type of vaccine, number of vaccine doses received, complete vaccination schedule, and having had COVID-19. Results: The vaccines received mostly were Vaxzevria® and Comirnaty® , with 73.3% between both, although 67.2% presented a complete regimen. The results of the OJABIO rapid neutralizing antibody test gave a positive result in 61.4% of the sample. There was a high association between the variables COVID-19 infection, two doses of vaccine, Spikevax® or Comirnaty® vaccine, and eighteen/twenty-nine years old group with a positive result on the OJABIO test. A total of 712 subjects answered the parallel survey (80%) on adverse effects and preferences between the different vaccines against COVID-19...(AU)

Pharmacogenetics May Prevent Psychotropic Adverse Events in Autism Spectrum Disorder: An Observational Pilot Study.

INTRODUCTION: Up to 73% of individuals with autism spectrum disorder (ASD) and intellectual disability (ID) currently have prescriptions for psychotropic drugs. This is explained by a higher prevalence of medical and psychiatric chronic comorbidities, which favors polypharmacy, increasing the probability of the appearance of adverse events (AEs). These could be a preventable cause of harm to patients with ASD and an unnecessary waste of healthcare resources. OBJECTIVE: To study the impact of pharmacogenetic markers on the prevention of AE appearance in a population with ASD and ID. METHODS: This is a cross-sectional, observational study (n = 118, 72 participants completed all information) in the ASD population. Sociodemographic and pharmacological data were gathered. The Udvalg for Kliniske Undersøgelser Scale (UKU Scale) was used to identify AEs related to the use of psychotropic medication. Polymorphisms of DOP2, ABCB1, and COMT were genotyped and correlated with the AE to find candidate genes. Furthermore, a review of all medications assessed in a clinical trial for adults with autism was performed to enrich the search for potential pharmacogenetic markers, keeping in mind the usual medications. RESULTS: The majority of the study population were men (75%) with multiple comorbidities and polypharmacy, the most frequently prescribed drugs were antipsychotics (69%); 21% of the participants had four or more AEs related to psychotropic drugs. The most common were "Neurological" and" Psychiatric" (both 41%). Statistical analysis results suggested a significant correlation between the neurological symptoms and the DOP2 genotype, given that they are not equally distributed among its allelic variants. The final review considered 19 manuscripts of medications for adults with ASD, and the confirmed genetic markers for those medications were consulted in databases. CONCLUSION: A possible correlation between neurologic AEs and polymorphisms of DOP2 was observed; therefore, studying this gene could contribute to the safety of this population's prescriptions. The following studies are underway to maximize statistical power and have a better representation of the population.

Detection of Neutralizing Antibodies against SARS-CoV-2 Post-Vaccination in Health Care Workers of a Large Tertiary Hospital in Spain by Using a Rapid Test LFIC and sVNT-ELISA.

The presence of neutralizing antibodies (NAbs) against SARS-CoV-2 represent a surrogate marker of immunologic protection in populations at high risk of infection such as healthcare workers caring for hospitalized patients with COVID-19. As recommended by CDC and the European CDC, the use of rapid diagnostic tests during population-based evaluations offers an opportunity to identify individuals with serologic evidence of natural infection or who have undergone vaccination. We carried out a cross-sectional study to assess the presence of neutralizing antibodies against SARS-CoV-2 among medical providers at an intensive care unit of a large referral hospital in Alicante, Spain. In addition, we tested for the presence of neutralizing antibodies compared to serum of uninfected individuals from a Biobank. We were also interested in evaluating the use of a rapid lateral flow immunochromatography (LFIC) test against a surrogate ELISA viral neutralization test (sVNT). This rapid test demonstrated a specificity of 1.000 95% CI (0.91-1.00) and the sensitivity of 0.987 95% CI (0.93-1.00). The negative predictive value was 95%. After six months, this rapid test demonstrated that those immunized with two doses of BioNTech/Pfizer vaccine, maintained optimal levels of neutralizing antibodies. We concluded that all Health Care Workers develop NAbs and the use of this rapid immunochromatographic test represents a potential tool to be used in population-based studies to detect serological antibody responses to vaccination. Vaccination policies could benefit from this tool to assess additional doses of vaccine or boosters among high-risk populations.

Knowledge, Attitudes, and Sources of Information on Vaccines in Spanish Nursing Students: A Cross-Sectional Study.

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.

Seroprevalence Study and Cross-Sectional Survey on COVID-19 for a Plan to Reopen the University of Alicante (Spain).

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.

Immunization Coverage of Inmates in Spanish Prisons.

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-diphtheria, it was 71.9%. However, for hepatitis A and pneumococcus infection, it was insignificant (<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.

Trends of Adverse Events Following Immunization (AEFI) Reports of Human Papillomavirus Vaccine in the Valencian Community-Spain (2008-2018).

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, -54.9%; 95%CI [-75.2; -17.7]), followed by a stable trend (-13% APC, 95%CI [-26.1; 2.4]). For the age group analysis, only the group aged 14-15 years old followed the same trend with -58.4% (95%CI [-73.9; -33.8]) APC during 2008-2011, and -8.8% (95%CI [-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 14-15 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.

Colorectal cancer molecular classification using BRAF, KRAS, microsatellite instability and CIMP status: Prognostic implications and response to chemotherapy.

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.

TFAP2E Methylation and Expression Status Does Not Predict Response to 5-FU-based Chemotherapy in Colorectal Cancer.

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.

KRAS and BRAF somatic mutations in colonic polyps and the risk of metachronous neoplasia.

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.

Increased Risk of Colorectal Cancer in Patients With Multiple Serrated Polyps and Their First-Degree Relatives.

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.

Serrated colorectal cancer: Molecular classification, prognosis, and response to chemotherapy.

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.

Endoscopic surveillance in patients with multiple (10-100) colorectal polyps.

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.

Immunohistochemical, genetic and epigenetic profiles of hereditary and triple negative breast cancers. Relevance in personalized medicine.

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.

Prevalence of MLH1 constitutional epimutations as a cause of Lynch syndrome in unselected versus selected consecutive series of patients with colorectal cancer.

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<0.0001). No constitutional epimutations in MLH1 were detected in the unselected population (0/62); five cases from the selected series were positive for MLH1 epimutations (15.6%, 5/32; p=0.004). CONCLUSIONS: Our results suggest a negligible prevalence of MLH1 constitutional epimutations in unselected cases of CRC. Therefore, MLH1 constitutional epimutation analysis should be conducted only for patients who fulfil the rBG and who lack MLH1 expression with methylated MLH1.

Methylation of tumor suppressor genes is related with copy number aberrations in breast cancer.

This study investigates the relationship of promoter methylation in tumor suppressor genes with copy-number aberrations (CNA) and with tumor markers in breast cancer (BCs). The study includes 98 formalin fixed paraffin-embedded BCs in which promoter methylation of 24 tumour suppressor genes were assessed by Methylation-Specific Multiplex Ligation-dependent Probe Amplification (MS-MLPA), CNA of 20 BC related genes by MLPA and ER, PR, HER2, CK5/6, CK18, EGFR, Cadherin-E, P53, Ki-67 and PARP expression by immunohistochemistry (IHC). Cluster analysis classed BCs in two groups according to promoter methylation percentage: the highly-methylated group (16 BCs), containing mostly hyper-methylated genes, and the sparsely-methylated group (82 BCs) with hypo-methylated genes. ATM, CDKN2A, VHL, CHFR and CDKN2B showed the greatest differences in the mean methylation percentage between these groups. We found no relationship of the IHC parameters or pathological features with methylation status, except for Catherin-E (p = 0.008). However the highly methylated BCs showed higher CNA proportion than the sparsely methylated BCs (p < 0.001, OR = 1.62; IC 95% [1.26, 2.07]). CDC6, MAPT, MED1, PRMD14 and AURKA showed the major differences in the CNA percentage between the two groups, exceeding the 22%. Methylation in RASSF1, CASP8, DAPK1 and GSTP1 conferred the highest probability of harboring CNA. Our results show a new link between promoter methylation and CNA giving support to the importance of methylation events to establish new BCs subtypes. Our findings may be also of relevance in personalized therapy assessment, which could benefit the hyper methylated BC patients group.

IGFBP3 methylation is a novel diagnostic and predictive biomarker in colorectal cancer.

BACKGROUND AND AIM: Aberrant hypermethylation of cancer-related genes has emerged as a promising strategy for the development of diagnostic, prognostic and predictive biomarkers in human cancer, including colorectal cancer (CRC). The aim of this study was to perform a systematic and comprehensive analysis of a panel of CRC-specific genes as potential diagnostic, prognostic and predictive biomarkers in a large, population-based CRC cohort. PATIENTS AND METHODS: Methylation status of the SEPT9, TWIST1, IGFBP3, GAS7, ALX4 and miR137 genes was studied by quantitative bisulfite pyrosequencing in a population-based cohort of 425 CRC patients. RESULTS: Methylation levels of all genes analyzed were significantly higher in tumor tissues compared to normal mucosa (p<0.0001); however, cancer-associated hypermethylation was most frequently observed for miR137 (86.7%) and IGFBP3 (83%) in CRC patients. Methylation analysis using the combination of these two genes demonstrated greatest accuracy for the identification of colonic tumors (sensitivity 95.5%; specificity 90.5%). Low levels of IGFBP3 promoter methylation emerged as an independent risk factor for predicting poor disease free survival in stage II and III CRC patients (HR = 0.49, 95% CI: 0.28-0.85, p = 0.01). Our results also suggest that stage II & III CRC patients with high levels of IGFBP3 methylation do not benefit from adjuvant 5FU-based chemotherapy. CONCLUSION: By analyzing a large, population-based CRC cohort, we demonstrate the potential clinical significance of miR137 and IGFBP3 hypermethylation as promising diagnostic biomarkers in CRC. Our data also revealed that IGFBP3 hypermethylation may serve as an independent prognostic and predictive biomarker in stage II and III CRC patients.

New insights into POLE and POLD1 germline mutations in familial colorectal cancer and polyposis.

Germline mutations in DNA polymerase ɛ (POLE) and δ (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.