Molecular study of the BRCA1 and BRCA2 genes in 153 breast cancer families from Castilla y León (Spain): new nine unclassified variants identified

[ES]: Fundamento: El cáncer de mama hereditario representa un 5-10% de todos los cánceres de mama. En la actualidad dos genes están asociados a la enfermedad, el BRCA1 y el BRCA2. Se sabe que mutaciones en estos genes aumentan el riesgo de padecer cáncer de mama hasta en un 80% en las portadoras. El objetivo de este estudio es la detección y caracterización de mutaciones en estos genes, en pacientes con cáncer de mama seleccionadas según criterios de moderado-alto riesgo pertenecientes a la Comunidad Autónoma de Castilla y León. Pacientes y método: Se analizaron 207 muestras seleccionadas pertenecientes a 153 familias. Se realizó extracción de ADN de sangre periférica y para la detección de mutaciones se emplearon técnicas de PCR múltiplex-heterodúplex-CSGE y secuenciación. Resultados: Se detectaron 45 cambios nucleotídicos distintos (23 en BRCA1 y 22 en BRCA2) en 74 familias (48,4% del total), que corresponden a 13 polimorfismos (29 familias), 19 variantes de efecto desconocido (26 familias), de las que 9 son descritas por primera vez en este trabajo, y 13 patológicas (19 familias; 12,42% de las familias). De las mutaciones patológicas, 8 (42,1%) afectan a BRCA1 y 11 (57,9%) a BRCA2. La mutación más frecuente es la 3036delACAA de BRCA2, presente en 4 familias no relacionadas. Conclusiones: El alto porcentaje de mutaciones, polimorfismos y variantes de efecto desconocido detectado revela la alta resolución del método de análisis mutacional utilizado, así como la validez de los criterios de selección aplicados. Existe un gran número de variantes de significado desconocido cuyo papel en la enfermedad debe ser clarificado mediante diferentes tipos de estudios, entre los que se incluye su tipificación en poblaciones control.[EN]: Background: It is estimated that 5-10% of breast cancers have an hereditary origin, germline mutations of BRCA1 and BRCA2 genes causing a predisposition. In the present study we analyzed BRCA1 and BRCA2 mutations in moderate to high risk breast cancer patients in order to find out the types and frequency of these mutations in the Spanish regional community of Castilla y León. Patients and method: We studied 207 moderate to high risk patients from 153 selected families. Genomic DNA was extracted from peripheral blood and analyzed by multiplex polymerase chain reaction-heteroduplexes-conformation sensitive gel electrophoresis (multiplex PCR-HA-CSGE). All variants detected were sequenced to further verify the mutation. Results: 45 alterations (23 in BRCA1 and 22 in BRCA2) were identified in 74 families (48.4%), corresponding to 13 polymorphisms (29 families), 19 unclassified variants (26 families) ­of which 9 have not been previously described­ and 13 cancer-prone mutations (19 families; 12.42% of all families). Eight out of the 19 deleterious mutations (42.1%) were detected in the BRCA1 gene and 11 (57.9%) in the BRCA2 gene. The most prevalent mutation was 3036delACAA, which was detected in four unrelated families. Conclusions: The high proportion of mutations, polymorphisms and unclassified variants we have detected may be the result of the sensitive procedure and the risk selection criteria used in this study. There is a high proportion of unclassified variants. Their role in the disease must be clarified through more studies, including their typing in control samples.Peer reviewe

Rapid mutation detection in complex genes by heteroduplex analysis with capillary array electrophoresis

14 páginas, 4 figuras, 3 tablas.Mutational analysis of large multiexon genes without prevalent mutations is a laborious undertaking that requires the use of a high-throughput scanning technique. The Human Genome Project has enabled the development of powerful techniques for mutation detection in large multiexon genes. We have transferred heteroduplex analysis (HA) by conformation-sensitive gel electrophoresis of the two major breast cancer (BC) predisposing genes, BRCA1 and BRCA2, to a multicapillary DNA sequencer in order to increase the throughput of this technique. This new method that we have called heteroduplex analysis by capillary array electrophoresis (HA-CAE) is based on the use of multiplex-polymerase chain reaction (PCR), different fluorescent labels and HA in a 16-capillary DNA sequencer. To date, a total of 114 different DNA sequence variants (19 insertions/deletions and 95 single-nucleotide substitutions – SNS) of BRCA1 and BRCA2 from 431 unrelated BC families have been successfully detected by HA-CAE. In addition, we have optimized the multiplex-PCR conditions for the colorectal cancer genes MLH1 and MSH2 in order to analyze them by HA-CAE. Both genes have been amplified in 13 multiplex groups, which contain the 35 exons, and their corresponding flanking intronic sequences. MLH1 and MSH2 have been analyzed in nine hereditary nonpolyposis colorectal cancer patients, and we have found six different DNA changes: one complex deletion/insertion mutation in MLH1 exon 19 and another five SNS. Only the complex mutation and one SNS may be classified as cancer-prone mutations. Our experience has revealed that HA-CAE is a simple, fast, reproducible and sensitive method to scan the sequences of complex genes.This work was supported by the Consejería de Sanidad (Junta y Castilla y León) through the regional Cancer Prevention Program, and by the ‘Fundación Burgos por la Investigación de la Salud’, Hospital General Yagüe, Burgos, Spain.Peer reviewe

Impacto de la corrosividad atmosférica sobre la infraestructura del sector y sobre los costos AOM

IP 1115-06-17398Objetivo general: Evaluar los impactos de la corrosividad del ambiente sobre la vida útil del SEC y sobre el factor AOM en la regulación, generando información y metodologías aplicables a las diferentes regiones del país donde se encuentra instalada dicha infraestructura y en regiones de futuro desarrollo y adicionalmente fortaleciendo la capacidad investigativas y el desarrollo tecnológico tanto de las entidades beneficiarias como de las ejecutora

Impact of INICC Multidimensional Hand Hygiene Approach in ICUs in Four Cities in Argentina

We evaluated the impact of the International Nosocomial Infection Control Consortium multidimensional approach to hand hygiene in 11 intensive care units in 4 cities in Argentina and analyzed predictors of poor hand hygiene compliance. We had a baseline period and a follow-up period. We observed 21 100 hand hygiene opportunities. Hand hygiene compliance increased from 28.3% to 64.8% (P = .0001). Males versus females (56.8% vs 66.4%; P < .001) and physicians versus nurses (46.6% vs 67.8%; P < .001) were significantly associated with poor hand hygiene compliance

Time-dependent analysis of extra length of stay and mortality due to ventilator-associated pneumonia in intensive-care units of ten limited-resources countries: findings of the International Nosocomial Infection Control Consortium (INICC)

Ventilator-associated pneumonias (VAPs) are a worldwide problem that significantly increases patient morbidity, mortality, and length of stay (LoS), and their effects should be estimated to account for the timing of infection. The purpose of the study was to estimate extra LoS and mortality in an intensive-care unit (ICU) due to a VAP in a cohort of 69 248 admissions followed for 283 069 days in ICUs from 10 countries. Data were arranged according to the multi-state format. Extra LoS and increased risk of death were estimated independently in each country, and their results were combined using a random-effects meta-analysis. VAP prolonged LoS by an average of 2.03 days (95% CI 1.52-2.54 days), and increased the risk of death by 14% (95% CI 2-27). The increased risk of death due to VAP was explained by confounding with patient morbidity