High-throughput mutation detection method to scan BRCA1 and BRCA2 based on heteroduplex analysis by capillary array electrophoresis

8 páginas, 3 figuras, 2 tablas.[Background]: Scanning for mutations in BRCA1 and BRCA2 in a large number of samples is hampered by the large sizes of these genes and the scattering of mutations throughout their coding sequences. Automated capillary electrophoresis has been shown to be a powerful system to detect mutations by either single-strand conformation polymorphism or heteroduplex analysis (HA). [Methods]: We investigated the adaptation of gel-based HA of BRCA1 and BRCA2 to a fluorescent multicapillary platform to increase the throughput of this technique. We combined multiplex PCR, three different fluorescent labels, and HA in a 16-capillary DNA sequencer and tested 57 DNA sequence variants (11 insertions/deletions and 46 single-nucleotide changes) of BRCA1 and BRCA2. [Results]: We detected all 57 DNA changes in a blinded assay, and 2 additional single-nucleotide substitutions (1186 A>G of BRCA1 and 3624 A>G of BRCA2), previously unresolved by conformation-sensitive gel electrophoresis. Furthermore, different DNA changes in the same PCR fragment could be distinguished by their peak patterns. [Conclusions]: Capillary-based HA is a fast, efficient, and sensitive method that considerably reduces the amount of “hands-on” time for each sample. By this approach, the entire coding regions of BRCA1 and BRCA2 from two breast cancer patients can be scanned in a single run of 90 min.This work was supported by the Consejería de Sanidad (Junta y Castilla y León) through the regional Breast Cancer Prevention Program.Peer reviewe

Propuesta de guion de cortometraje de ficción basado en el perfil arrojado por los estudiantes que sufren de insomnio no orgánico del quinto al octavo ciclo de la carrera de comunicación de la Universidad Católica Santo Toribio de Mogrovejo del semestre 2018 – II

La presente tesis titulada “Propuesta de guion de cortometraje de ficción basado en el perfil arrojado por los estudiantes que sufren de insomnio no orgánico del quinto al octavo ciclo de la carrera de Comunicación de la Universidad Católica Santo Toribio de Mogrovejo del semestre 2018 - II”, pretende profundizar en el estudio del guion a partir de un perfil y sus requisitos necesarios pues este es la base para generar un producto audiovisual que nos permita contar una historia, específicamente, la del insomnio no orgánico en estudiantes universitarios. Esta investigación tuvo como objetivos específicos identificar los casos de insomnio no orgánico en los estudiantes del quinto al octavo ciclo de la carrera de Comunicación de la Universidad Católica Santo Toribio de Mogrovejo. Luego, se procedió a entrevistar a expertos en psicología y psiquiatría con el fin de saber las características en común de los casos y así, posteriormente, elaboraron un perfil con la información recopilada a los estudiantes. Con toda esa información se escribió un guion basado en la estructura dada por los aspectos teóricos y el perfil. Por último, se consultó a guionistas del medio para las posibles correcciones en la propuesta de guion. Los instrumentos aplicados fueron: entrevistas a expertos de insomnio no orgánico, además de un test insomnio otorgado por ellos mismos para realizarlo a los estudiantes y cuestionarios de opinión

Modification of stool's water content in constipated infants: management with an adapted infant formula

<p>Abstract</p> <p>Background</p> <p>Constipation is a common occurrence in formula-fed infants. The aim of this preliminary study was to evaluate the impact of a formula with high levels of lactose and magnesium, in compliance with the official regulations, on stool water content, as well as a parental assessment of constipation.</p> <p>Materials and methods</p> <p>Thirty healthy term-born, formula-fed infants, aged 4-10 weeks, with functional constipation were included. All infants were full-term and fed standard formula. Exclusion criteria were preterm and/or low birth weight, organic constipation, being breast fed or fed a formula specially designed to treat constipation. Stool composition was measured by near-infrared reflectance analysis (NIRA) and parents answered questions about crying associated with defecation and stool consistency at baseline and after two weeks of the adapted formula.</p> <p>Results</p> <p>After 2 weeks of the adapted formula, stool water content increased from 71 +/- 8.1% to 84 +/- 5.9%, (p < 0.02). There was no significant change in the stool's fat, protein or carbohydrate content. Parental impressions of constipation were improved with the decrease in stool hardness (100% with hard stools at baseline, 10% after 2 weeks), pain with defecation (90% at baseline, 10% after 2 weeks), and the requirement for rectal stimulation to achieve defecation (70% at baseline, 30% after 2 weeks, p < 0.001 for all three indicators).</p> <p>Conclusions</p> <p>This preliminary study suggests that an adapted formula with high levels of lactose and magnesium increases stool water content and improves symptoms of constipation in term-born, formula-fed infants. A larger randomized placebo-controlled trial is indicated.</p

Analysis of PALB2 gene in BRCA1/BRCA2 negative Spanish hereditary breast/ovarian cancer families with pancreatic cancer cases

This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al.[Background]: The PALB2 gene, also known as FANCN, forms a bond and co-localizes with BRCA2 in DNA repair. Germline mutations in PALB2 have been identified in approximately 1% of familial breast cancer and 3–4% of familial pancreatic cancer. The goal of this study was to determine the prevalence of PALB2 mutations in a population of BRCA1/BRCA2 negative breast cancer patients selected from either a personal or family history of pancreatic cancer. [Methods]: 132 non-BRCA1/BRCA2 breast/ovarian cancer families with at least one pancreatic cancer case were included in the study. PALB2 mutational analysis was performed by direct sequencing of all coding exons and intron/exon boundaries, as well as multiplex ligation-dependent probe amplification. [Results]: Two PALB2 truncating mutations, the c.1653T>A (p.Tyr551Stop) previously reported, and c.3362del (p.Gly1121ValfsX3) which is a novel frameshift mutation, were identified. Moreover, several PALB2 variants were detected; some of them were predicted as pathological by bioinformatic analysis. Considering truncating mutations, the prevalence rate of our population of BRCA1/2-negative breast cancer patients with pancreatic cancer is 1.5%. [Conclusions]: The prevalence rate of PALB2 mutations in non-BRCA1/BRCA2 breast/ovarian cancer families, selected from either a personal or family pancreatic cancer history, is similar to that previously described for unselected breast/ovarian cancer families. Future research directed towards identifying other gene(s) involved in the development of breast/pancreatic cancer families is required.This research was supported by grants from the Xunta de Galicia (10PXIB 9101297PR) and FMM Foundation given to AV. MH was supported from the Instituto de Salud Carlos III, Fondo de Investigación Sanitaria (FIS) Research Grant 09/00859, and Fundación Mutua Madrileña (FMM) Research Grant FMM-08. SGE is supported by a Miguel Servet contract of the Instituto de Salud Carlos III. EAV was supported in part by grants BIO39/VA27/10 (Consejería de Sanidad) and CSI004A10-2 (Consejería de Educación) from the Junta de Castilla y León.Peer Reviewe

Multibranched acquired digital fibrokeratoma. Clinical case

El fibroqueratoma es una lesión tumoral benigna del aparato ungueal. Se trata de una lesión adquirida que se desarrolla a partir de un collarete epidérmico queratósico que rodea su base. Son tumores cutáneos, de naturaleza benigna, asintomáticos, generalmente de pequeño tamaño, consistencia firme, encapsulados y con una capa superficial gris perlada. Tienen una etiología desconocida. Morfológicamente, aparecen como una neoformación de forma semiesférica recubierta completamente por piel normal sonrosada y, generalmente, son asintomáticos. Inusualmente, los podemos encontrar como lesiones multilobuladas o multirramificadas. El diagnóstico es fundamentalmente clínico, apoyado en el informe de anatomía patológica de la lesión tras su escisión quirúrgica. El presente artículo presenta un caso clínico de escisión quirúrgica de un fibroqueratoma mostrándolo como una buena alternativa de tratamiento que ofrece una solución definitiva, sencilla y sin complicaciones para el paciente.Fibrokeratoma is a benign tumor of the nail. It is an acquired injury that develops from a keratotic epidermal collar that surrounds its base. They are benign, asymptomatic skin tumors, generally small in size, firm in consistency, encapsulated and with a pearly gray surface layer. They have an unknown etiology. Morphologically, they appear as a hemispherical neoformation completely covered by normal skin and are generally asymptomatic. Unusually, we can find them as multi-lobed or multi-branched lesions. Its diagnosis is mainly clinical, supported by the pathological report of the lesion after its surgical excision. The present paper presents a clinical case of a surgical excision of a fibrokeratoma showing it as a good treatment alternative that offers a definitive, simple and uncomplicated solution for the patient

GALNT12 is Not a Major Contributor of Familial Colorectal Cancer Type X

Previous evidence indicates that mutations in the GALNT12 gene might cause a fraction of the unexplained familial colorectal cancer (CRC) cases: GALNT12 is located in 9q22-33, in close proximity to a CRC linkage peak; and germline missense variants that reduce the enzymatic activity of the protein have been identified in CRC patients, some of them with familial CRC history. We hypothesized that mutations in GALNT12 might explain part of the high-risk families grouped as familial CRC type X (fCRC-X), that is, Amsterdam-positive families with mismatch repair proficient tumors. We sequenced the coding regions of the gene in 103 probands of fCRC-X families, finding no functionally relevant mutations. Our results rule out GALNT12 as a major high CRC susceptibility gene. Additional studies are required to provide further evidence about its role as a moderate/low susceptibility gene in familial aggregation of cancer

Germline Genetic Findings Which May Impact Therapeutic Decisions in Families with a Presumed Predisposition for Hereditary Breast and Ovarian Cancer

In this study, we aim to gain insight in the germline mutation spectrum of ATM, BARD1, BRIP1, ERCC4, PALB2, RAD51C and RAD51D in breast and ovarian cancer families from Spain. We have selected 180 index cases in whom a germline mutation in BRCA1 and BRCA2 was previously ruled out. The importance of disease-causing variants in these genes lies in the fact that they may have possible therapeutic implications according to clinical guidelines. All variants were assessed by combined annotation dependent depletion (CADD) for scoring their deleteriousness. In addition, we used the cancer genome interpreter to explore the implications of some variants in drug response. Finally, we compiled and evaluated the family history to assess whether carrying a pathogenic mutation was associated with age at diagnosis, tumour diversity of the pedigree and total number of cancer cases in the family. Eight unequivocal pathogenic mutations were found and another fourteen were prioritized as possible causal variants. Some of these molecular results could contribute to cancer diagnosis, treatment selection and prevention. We found a statistically significant association between tumour diversity in the family and carrying a variant with a high score predicting pathogenicity (p = 0.0003)

Germline genetic findings which may impact therapeutic decisions in families with a presumed predisposition for hereditary breast and ovarian cancer

In this study, we aim to gain insight in the germline mutation spectrum of ATM, BARD1, BRIP1, ERCC4, PALB2, RAD51C and RAD51D in breast and ovarian cancer families from Spain. We have selected 180 index cases in whom a germline mutation in BRCA1 and BRCA2 was previously ruled out. The importance of disease-causing variants in these genes lies in the fact that they may have possible therapeutic implications according to clinical guidelines. All variants were assessed by combined annotation dependent depletion (CADD) for scoring their deleteriousness. In addition, we used the cancer genome interpreter to explore the implications of some variants in drug response. Finally, we compiled and evaluated the family history to assess whether carrying a pathogenic mutation was associated with age at diagnosis, tumour diversity of the pedigree and total number of cancer cases in the family. Eight unequivocal pathogenic mutations were found and another fourteen were prioritized as possible causal variants. Some of these molecular results could contribute to cancer diagnosis, treatment selection and prevention. We found a statistically significant association between tumour diversity in the family and carrying a variant with a high score predicting pathogenicity (p = 0.0003)

The highly prevalent BRCA2 mutation c.2808_2811del (3036delACAA) is located in a mutational hotspot and has multiple origins

BRCA2-c.2808_2811del (3036delACAA) is one of the most reported germ line mutations in non-Ashkenazi breast cancer patients. We investigated its genetic origin in 51 Spanish carrier families that were genotyped with 11 13q polymorphic markers. Three independent associated haplotypes were clearly distinguished accounting for 23 [west Castilla y León (WCL)], 20 [east Castilla y León (ECL)] and 6 (South of Spain) families. Mutation age was estimated with the Disequilibrium Mapping using Likelihood Estimation software in a range of 45–68 and 45–71 generations for WCL and ECL haplotypes, respectively. The most prevalent variants, c.2808_2811del and c.2803G > A, were located in a double-hairpin loop structure (c.2794–c.2825) predicted by Quikfold that was proposed as a mutational hotspot. To check this hypothesis, random mutagenesis was performed over a 923 bp fragment of BRCA2, and 86 DNA variants were characterized. Interestingly, three mutations reported in the mutation databases (c.2680G > A, c.2944del and c.2957dup) were replicated and 20 affected the same position with different nucleotide changes. Moreover, five variants were placed in the same hairpin loop of c.2808_2811del, and one affected the same position (c.2808A > G). In conclusion, our results support that at least three different mutational events occurred to generate c.2808_2811del. Other highly prevalent DNA variants, such as BRCA1-c.68_69delAG, BRCA2- c.5946delT and c.8537delAG, are concentrated in hairpin loops, suggesting that these structures may represent mutational hotspots