Study of Genetic Susceptibility to Age-related Macular Degeneration

Age-related macular degeneration is the leading cause of severe and irreversible central vision loss in people over 50 in developed countries. This disease can be classified into two groups, wet and dry, the latter being the object of study in this paper, which is characterized by the development of choroidal neovascularization. Due to the importance of SNPs in the development of this disease, three polymorphisms belonging to the gene that encodes endogline (rs1998923, rs1330683 and rs1330684) and a polymorphism belonging to the gene that encodes preseniline-1 (rs165932) will be studied. Therefore, the objective of this work is to determine if the development of this disease is related to these polymorphisms. For this purpose, a total of 326 DNA samples were selected from patients at the University Hospital of Salamanca, of which 175 were diagnosed with wet age-related macular degeneration (cases) and 151 with cataracts (controls). The different genotypes were analysed using allelic discrimination with Taqman® probes. After performing the chi-squared test to check whether there was a statistically significant difference (p < 0.05), it is concluded that the development of wet age-related macular degeneration will not be related to these polymorphisms.La degeneración macular asociada a la edad es la principal causa de pérdida de visión central severa e irreversible en personas mayores de 50 años en los países desarrollados./nEsta enfermedad se puede clasificar en dos grupos, seca y húmeda, siendo esta última objeto de estudio en este trabajo, la cual se caracteriza por el desarrollo de neovascularización coroidea. Debido a la importancia de los SNPs en el desarrollo de esta enfermedad, se estudiarán tres polimorfismos pertenecientes al gen que codifica la endoglina (rs1998923, rs1330683 y rs1330684) y un polimorfismo perteneciente al gen que codifica la presenilina-1 (rs165932). Por tanto, el objetivo de este trabajo es determinar si el desarrollo de esta enfermedad está relacionado con estos polimorfismos. Para ello, se seleccionaron un total de 326 muestras de ADN de pacientes del Hospital Universitario de Salamanca, de los cuales 175 estaban diagnosticados de DMAE húmeda (casos) y 151 de cataratas (controles) y se analizaron los distintos genotipos mediante discriminación alélica con sondas Taqman®. Tras realizar el test chi-cuadrado para comprobar si existía una diferencia estadísticamente significativa (p < 0.05), se concluye que el desarrollo de degeneración macular asociada a la edad húmeda no va a estar relacionado con estos polimorfismos

Evaluation of chloroquine effect in autophagy of breast cancer cells and its possible use in chemotherapy

Autophagy is a celular mechanism which is in charge of destroying damaged organelles and obtaining nutrients in fasting periods. This process has a very important role in cancer regulation. In early stages of cancer, autophagy prevents the growth of tumoral cells. However, in final stages, it promotes metastasis and resistance to chemotherapy. Chloroquine is an antipaludic drug which, according to recent research, might have antineoplasic properties due to inhibition of autophagy. The target of this research is to prove, in one hand, the citotoxic effect of chloroquine in a breast cancer cell line (BT-549) by performing a MTT assay; and, in the other hand, to study the effect of chloroquine in the expression of three autophagic proteins (LC3, p62 and Beclin-1) using a Western Blot. The results indicate that chloroquine inhibites the growth of tumoral cells at 50 µM and that chloroquine increases the expression of the proteins, which could indicate that chloroquine is inhibiting autophagy. In conclusion, chloroquine might be, in the future,  a useful drug in breast cancer chemotherapy. La autofagia es un proceso celular que se encarga de la degradación de orgánulos dañados y de la obtención de nutrientes en condiciones de ayuno. Este proceso tiene un papel muy importante en la regulación del cáncer. En fases iniciales impide el desarrollo de células tumorales; mientras que, en fases finales del cáncer, promueve las metástasis y la resistencia a la quimioterapia. Cloroquina es un fármaco antipalúdico al que, en estudios recientes, se le han atribuido propiedades antineoplásicas debido a un mecanismo de inhibición de la autofagia. El objetivo de este trabajo es comprobar, por un lado, el efecto citotóxico de la cloroquina sobre la línea celular BT-549 de cáncer de mama mediante un ensayo MTT; y por otro, estudiar el efecto de la cloroquina sobre tres proteínas autofágicas: LC3, p62 y Beclin-1 por medio de un Western Blot. Los resultados obtenidos indican que la cloroquina inhibe el crecimiento celular a 50 µM y que la cloroquina aumenta la expresión de las proteínas estudiadas, lo que se podría considerar un mecanismo compensador de la célula ante la inhibición de la autofagia producida por cloroquina. En conclusión, la cloroquina podría ser un fármaco con utilidad en el tratamiento del cáncer de mama

COL4A4 gene study of a European population: description of new mutations causing autosomal dominant Alport syndrome

[Background]: Autosomal forms of Alport syndrome represent 20% of all patients (15% recessive and 5% dominant). They are caused by mutations in the COL4A3 and COL4A4 genes, which encode a-3 and a-4 collagen IV chains of the glomerular basement membrane, cochlea and eye. Thin basement membrane nephropathy may affect up to 1% of the population. The pattern of inheritance in the 40% of cases is the same as autosomal dominant Alport syndrome: heterozygous mutations in these genes. The aim of this study is to detect new pathogenic mutations in the COL4A4 gene in the patients previously diagnosed with autosomal Alport syndrome and thin basement membrane nephropathy in our hospital. [Methods]: We conducted a clinical and genetic study in eleven patients belonging to six unrelated families with aforementioned clinical symptoms and a negative study of COL4A3 gene. The molecular study was made by conformation of sensitive gel electrophoresis (CSGE) and direct sequencing of the fragments that show an altered electrophoretic migration pattern. [Results]: We found two pathogenic mutations, not yet described: IVS3 + 1G > C is a replacement of Guanine to Cytosine in position +1 of intron 3, in the splicing region, which leads to a pathogenic mutation. c.4267C > T; p.P1423S is a missense mutation, also considered pathogenic. We also found seven new polymorphisms. [Conclusions]: We describe two new pathogenic mutations, responsible for autosomal dominant Alport syndrome. The other families of the study were undiagnosed owing to problems in the method employed and the possibility of mutations in other genes, giving rise to other diseases with similar symptoms.Peer Reviewe

Síndrome de Gorlin

[ES]El síndrome de Gorlin es una enfermedad infrecuente de herencia autosómica dominante producida por mutaciones en genes de la vía de señalización Sonic Hedgehog, entre los que destaca PTCH1. Se caracteriza por el desarrollo de múltiples carcinomas basocelulares en edades tempranas, que pueden ir asociados a otras manifestaciones cutáneas como pits palmoplantares, o a manifestaciones extracutáneas, entre las que destacan los queratoquistes odontogénicos y el meduloblastoma. El papel del dermatólogo es importante en la sospecha de este síndrome, pero suele ser necesario un equipo multidisciplinar en el diagnóstico, seguimiento y en el tratamiento de estos pacientes. El tratamiento dermatológico puede ser complicado debido al alto número de carcinomas basocelulares y a su extensión. En los últimos años se han desarrollado nuevos fármacos que inhiben la vía Sonic Hedgehog y parecen prometedores para estos pacientes, aunque su eficacia está limitada por los efectos secundarios y la creación de resistencias.[EN]Gorlin syndrome is a rare autosomal dominant disease caused by mutations in the sonic hedgehog signaling pathway. Of particular importance is the PTCH1 gene. The disease is characterized by the development of multiple basal cell carcinomas at young ages. These tumors may present with other skin manifestations such as palmoplantar pits and with extracutaneous manifestations such as odontogenic keratocysts and medulloblastoma. Although the dermatologist may be key for recognizing clinical suspicion of the syndrome, a multidisciplinary team is usually necessary for diagnosis, treatment, and follow-up. Skin treatment may be complicated due to the large number of basal cell carcinomas and the extent of involvement

The T309G MDM2 gene polymorphism is a novel risk factor for proliferative vitreoretinopathy

This is an open-access article distributed under the terms of the Creative Commons Attribution License.-- et al.Proliferative vitreoretinopathy (PVR) is still the major cause of failure in retinal detachment (RD) surgery. It is believed that down-regulation in the p53 pathway could be an important key in PVR pathogenesis. The purpose was to evaluate the impact of T309G MDM2 polymorphism (rs2279744) in PVR. Distribution of T309G MDM2 genotypes among European subjects undergoing RD surgery was evaluated. Proportions of genotypes between subsamples from different countries were analyzed. Also, a genetic interaction between rs2279744 in MDM2 and rs1042522 in p53 gene was analyzed. Significant differences were observed comparing MDM2 genotype frequencies at position 309 of intron 1 between cases (GG: 21.6%, TG: 54.5%, TT: 23.8%) and controls (GG: 7.3%, TG: 43.9%, TT: 48.7%). The proportions of genotypes between sub-samples from different countries showed a significant difference. Distribution of GG genotype revealed differences in Spain (35.1-53.0)/(22.6-32.9), Portugal (39.0-74.4)/(21.4-38.9), Netherlands (40.6-66.3)/(25.3-38.8) and UK (37.5-62.4)/(23.3-34.2). The OR of G carriers in the global sample was 5.9 (95% CI: 3.2 to 11.2). The OR of G carriers from Spain and Portugal was 5.4 (95% CI: 2.2-12.7), whereas in the UK and the Netherlands was 7.3 (95% CI: 2.8-19.1). Results indicate that the G allele of rs2279744 is associated with a higher risk of developing PVR in patients undergoing a RD surgery. Further studies are necessary to understand the role of this SNP in the development of PVR.This work was supported by SAF 2007-66394, FIS PI10/00219 and Group of Excellence Grant (GR15) from Junta de Castilla y León.Peer Reviewe

Methoxy and bromo scans on N-(5-methoxyphenyl) methoxybenzenesulphonamides reveal potent cytotoxic compounds, especially against the human breast adenocarcinoma MCF7 cell line

Thirty seven N-(5-methoxyphenyl)-4-methoxybenzenesulphonamide with methoxy or/and bromo substitutions (series 1-4) and with different substituents on the sulphonamide nitrogen have been synthesised. 21 showed sub-micromolar cytotoxicity against HeLa and HT-29 human tumour cell lines, and were particularly effective against MCF7. The most potent series has 2,5-dimethoxyanilines, especially the 4-brominated compounds 23–25. The active compounds inhibit microtubular protein polymerisation at micromolar concentrations, thus pointing at tubulin as the target. Co-treatment with the MDR inhibitor verapamil suggests that they are not MDR substrates. Compound 25 showed nanomolar antiproliferative potency. It severely disrupts the microtubule network in cells and arrests cells at the G2/M cell-cycle phase, thus confirming tubulin targeting. 25 triggered apoptotic cell death, and induced autophagy. Docking studies suggest binding in a distinct way to the colchicine site. These compounds are promising new antitumor agents acting on tubulin.We thank the people at Frigoríficos Salamanca S.A. slaughterhouse for providing us with the calf brains, “Servicio General de NMR” and “Servicio General de Espectrometría de Masas” of the University of Salamanca for equipment. M.G. acknowledges a predoctoral fellowship from the Junta de Castilla y León (ORDEN EDU/529/2017 de 26 de junio). M.O.-S. acknowledges a predoctoral fellowship from the IBSAL (IBpredoc17/00010). A.V.-B. acknowledges a predoctoral fellowship from the Spanish Ministerio de Educación, Cultura y Deporte (FPU15/02457). This research was funded by the Consejería de Educación de la Junta de Castilla y León (ORDEN EDU/529/2017 de 26 de junio, SA030U16, SA262P18 and SA116P20), co-funded by the EU’s European Regional Development Fund-FEDER, the Spanish Ministry of Science, Innovation and Universities (RTI2018-099474-BI00) and the health research program of the Instituto de Salud Carlos III (Spanish Ministry of Economy and Competitiveness) [PI16/01920 and PI20/01569] co-funded with FEDER funds. Ministerio de Educación, Cultura y Deporte [FPU15/02457], IBSAL [IBpredoc17/00010]

Cognitive outcome and gamma noise power unrelated to neuregulin 1 and 3 variation in schizophrenia

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.[Background]: Neuregulins are a family of signalling proteins that orchestrate a broad range of cellular responses. Four genes encoding Neuregulins 1-4 have been identified so far in vertebrates. Among them, Neuregulin 1 and Neuregulin 3 have been reported to contribute to an increased risk for developing schizophrenia. We hypothesized that three specific variants of these genes (rs6994992 and rs3924999 for Neuregulin 1 and rs10748842 for Neuregulin 3) that have been related to this illness may modify information processing capacity in the cortex, which would be reflected in electrophysiological parameters (P3b amplitude or gamma noise power) and/or cognitive performance. [Methods]: We obtained DNA from 31 patients with schizophrenia and 23 healthy controls and analyzed NRG1 rs6994992, NRG1 rs3924999 and NRG3 rs10748842 promoter polymorphisms by allelic discrimination with real-time polymerase chain reaction (PCR). We compared cognitive outcome, P300 amplitude parameters and an electroencephalographic measure of noise power in the gamma band between the groups dichotomized according to genotype. [Results]: Contrary to our hypothesis, we could not detect any significant influence of variation in Neuregulin 1/Neuregulin 3 polymorphisms on cognitive performance or electrophysiological parameters of patients with schizophrenia. [Conclusions]: Despite our findings, we cannot discard that other genetic variants and, more likely, interactions between those variants and with genetic variation related to different pathways may still influence cerebral processing in schizophrenia.Funding for this study was provided by the Instituto de Salud Carlos III Grants 080017 and 1102203 to VM, Gerencia Regional de Salud de Castilla y León GRS 249/A/08 and 613/A/11, a postdoctoral Marie Curie Intra European Fellowship within the 7th European Commission Framework Programme for Research and Innovation (330156-CODIP) to ÁD, a predoctoral research grant from the Consejería de Educación - Junta de Castilla y León and the European Social Fund to ÁD (EDU/1486/2008) and CC (EDU/1064/2009), a predoctoral scholarship from the University of Salamanca and Santander Bank to VS, and the FIS Grant PI 1000219 to RG.Peer Reviewe

ncRNA Study in Males with Breast Cancer

Male breast cancer (BC) is a rare disease accounting for less than 1% of all BC detected. For this reason, it would be important to find ways to prevent this kind of cancer, to know how it develops in men and if there is any genetic predisposition to have BC. We know that noncoding RNA (ncRNA) are involved in female breast cancer but there is no information about their role in males. In this paper we try to find if some ncRNA have implications in male BC. Specifically, we have studied some single nucleotide polymorphisms of the genes encoding for mir-146A, mir-196A2, mir-499A and long noncoding RNA, HotAIR. The technique used was genotyping with taqman probe, that allows to discriminate between different polymorphisms. The genetic counsil of cancer research center provides us with the study population. The preliminary results showed that the allele C of the polymorphism with rs2910164 of the mir-146A gene and the genotype CA of the polymorphism with rs1899663 of the HotAIR gene could protect males from developing BC.[ES] El cáncer de mama (CM) en varones es una enfermedad rara, ya que se trata de menos del 1% de los cánceres de mama detectados. Por ello es importante encontrar formas de prevenir este tipo de enfermedades y saber por qué se producen y si existe algún factor genético que predispone a padecer este tipo de cáncer. En este sentido, se sabe que algunos RNA no codificantes (ncRNA) están involucrados en el CM en mujeres, pero no hay nada descrito referente a su implicación en varones. En este trabajo se pretende averiguar si determinados ncRNA predisponen o protegen a varones de padecer CM. En concreto se han estudiado algunos polimorfismos de nucleótido único de los genes que codifican para los mir-146A, mir-196A2, mir-499A y el lnRNA HotAIR mediante la técnica de genotipado con sondas Taqman, que permite discriminar entre los polimorfismos de cada ncRNA. La población de estudio ha sido facilitada por el Consejo genético del Centro de Investigación del Cáncer. Los resultados obtenidos indican que el alelo C del polimorfismo rs2910164 del gen que codifica el mir-146A y el genotipo CA del polimorfismo rs1899663 del gen que codifica HOTAIR podrían proteger frente a la aparición de CM en varone

Interleukin-4 (IL4) and Interleukin-4 receptor (IL4RA) polymorphisms in asthma: a case control study

BACKGROUND: IL4/IL4RA pathway plays an important role in atopy and asthma. Different polymorphisms in IL4 and IL4RA genes have been described. Particularly, -33C>TIL4 and 576Q>RIL4RA SNPs have been independently associated to atopy and asthma. The purpose of this study was to analyse these polymorphisms in a population of patients with a well-characterized asthma phenotype. METHODS: A total of 212 unrelated Caucasian individuals, 133 patients with asthma and 79 healthy subjects without symptoms or history of asthma or atopy and with negative skin prick tests were recruited. Lung function was measured by spirometry and asthma was specialist physician-diagnosed according to the ATS (American Thoracic Society) criteria and classified following the GINA (Global Initiative for Asthma) guidelines. Skin prick tests were performed according to EAACI recommendations. -33C>TIL4 was studied with TaqMan assay and 576Q>RIL4RA by PCR-RFLP technique. Hardy-Weinberg equilibrium was analysed in all groups. Dichotomous variables were analysed using χ(2), Fisher exact test, Monte Carlo simulation test and odds ratio test. To model the effects of multiple covariates logistic regression was used. RESULTS: No statistically significant differences between the group of patients with asthma and the controls were found when the allele and genotype distribution of -33C>TIL4 and 576Q>RIL4RA polymorphisms were compared. However, the T allele of the -33C>TIL4 SNP was more frequent in patients with persistent asthma. Multivariate analysis adjusted for age and sex confirmed that carriers of allele T had an increased risk of persistent asthma (OR:2.77, 95%CI:1.18–6.49; p = 0.019). Analysis of combination of polymorphisms showed that patients carrying both the T allele of -33C>TIL4 and the A allele of 576Q>RIL4RA had an increased risk of asthma. This association was particularly observed in persistent asthma [Fisher's p value = 0.0021, Monte Carlo p value (after 10(4 )simulations) = 0.0016, OR:3.39; 95% CI:1.50–7.66]. CONCLUSION: Our results show a trend of association between the genetic combination of the T allele of -33C>TIL4 and the A allele of 576Q>RIL4RA with asthma. This genetic variant was more frequently observed in patients with persistent asthma. As long as this study was performed in a small population, further studies in other populations are needed to confirm these results