Proteomic Analysis in Seminal Plasma of Fertile Donors and Infertile Patients with Sperm DNA Fragmentation

Seminal plasma proteomics studies could represent a new approach for the determination of molecular elements driving male infertility, resulting in a better male infertility characterization. The aim of this study is to investigate proteomic differences in seminal plasma samples from fertile and infertile individuals. For that, semen samples were selected according to semen analysis, clinical pathology, and values of sperm DNA fragmentation (alkaline and neutral Comet assay and Sperm Chromatin Dispersion test). A total of 24 seminal plasma samples classified in four groups were processed: fertile donors (FD), recurrent miscarriage patients (RM), asthenoteratozoospermic patients (ATZ), and asthenoteratozoospermic patients with varicocele (ATZ-VAR). Results obtained by 2D-differential gel electrophoresis (2D-DIGE) revealed 26 spots significantly increased in fertile donors when compared to patient groups. Also, eight spots in the ATZ group and two in the ATZ-VAR group were decreased compared to the other groups. Twenty-eight proteins were identified by mass spectrometry (MS), most of them involved in metabolic and cellular processes and with a catalytic or binding function. Protein-protein interactions through Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) tool suggest that a large part of them were associated with each other. Furthermore, most of them were associated with ubiquitin C, indicating that it could play an important regulation role, resulting in a potential male infertility biomarker

Open access simulation toolbox for the grid connection of offshore wind farms using multi-terminal HVDC networks

Decarbonisation of the European electricity system can become dauntingly costly due to transmission and distribution network issues arising from the integration of intermittent renewable generation sources. It is expected that wind energy will be the principal renewable source by 2050 and, as such, a number of initiatives in the academia and in the industry are being carried out to propose solutions to best accommodate the wind resource. This paper presents work carried out by DEMO 1 partners within the EU FP7 project BEST PATHS. A MATLAB/Simulink toolbox consisting of the necessary building blocks for the simulation and integration of offshore wind farms using enabling technologies such as multiterminal high-voltage direct-current grids is presented. To illustrate the toolbox capabilities, a number of system topologies is studied. System performance is assessed and measured against a set of key performance indicators. To ensure knowledge dissemination, the toolbox has been made available as open access in the BEST PATHS project website

HIV/STI co-infection among men who have sex with men in Spain

In Spain, neither the HIV nor the STI national surveillance systems collect information on HIV/STI co-infection. However, there are two networks based on HIV/STI clinics which gather this data. We describe HIV prevalence in men who have sex with men (MSM) diagnosed with infectious syphilis and/or gonorrhoea in 15 STI clinics; and concurrent diagnoses of STI in MSM newly diagnosed with HIV in 19 HIV/STI clinics. In total, 572 MSM were diagnosed with infectious syphilis and 580 with gonorrhoea during 2005-2007. HIV prevalence among syphilis and gonorrhoea cases was 29.8% and 15.2% respectively. In the multivariate analysis, HIV/syphilis co-infection was associated with being Latin American; having a history of STI; reporting exclusively anal intercourse; and having sex with casual or several types of partners. HIV and gonorrhoea co-infection was associated with age older than 45 years; having no education or only primary education completed; and having a history of STI. In total, 1,462 HIV infections were newly diagnosed among MSM during 2003-2007. Of these, 31.0% were diagnosed with other STI at the same time. Factors associated with STI co-infection among new HIV cases in MSM were being Latin American; and having sex with casual partners or with both steady and casual partners. In Spain, a considerable proportion of MSM are co-infected with HIV and STI.This work was funded by two grants (36646/07; 36794/08) from the Foundation for Research and Prevention of AIDS in Spain (Fundación para la Investigación y la Prevención del SIDA en España–FIPSE).S

Increasing penetration of renewals in isolated power systems using energy storage systems

Abstract. Frequency stability is one of the most relevant issues in operation of isolated power systems. High penetration of renewals may affect significantly frequency stability of isolated power systems since wind and solar photovoltaic generation have neither inertia nor primary frequency regulation. An alternative is the use of storage systems in the most critical operating conditions in such a way that if a generator trips, storage systems are subsequently tripped and the power balance is restored. This paper presents an approach to determine the maximum renewable generation in an isolated power system to prevent non-admissible frequency excursions in case of generator tripping. Moreover, such approach allows sizing an energy storage system to increase the penetration of renewals

Inestabilidad cromosómica y desequilibrios genómicos en cáncer de vejiga

Los carcinomas uroteliales de vejiga, al igual que la mayoría de tumores sólidos, se caracterizan por la acumulación de múltiples desequilibrios genéticos. Con el fin de contribuir al conocimiento de las bases genéticas de la tumorogénesis urotelial, se estudió una serie de 180 tumores de vejiga mediante CGH convencional, observando que estos tumores mantienen un perfil característico de desequilibrios genómicos con ganancias en 1q, 8q, 11q, 16p, 17q, 18p, 19, 20q, Xq y pérdidas en 4q, 8p, 9p, 9q, 11p, 11q, 13q, 18q e Y. Las ganancias en 5p, 6p, 10p y las pérdidas en 5q, 6q aumentan su frecuencia de forma significativa a lo largo de la progresión tumoral. Estos resultados se confirmaron mediante la técnica de CGH array. Demostrando además, que los tumores músculo infiltrantes con desequilibrios en el cromosoma 5 muestran frecuentes amplificaciones en 1q, 5p, 6p y 19q, afectando entre otros a los genes CKS1, SKP2, E2F3 y CCNE1, que participan en la misma ruta de control del ciclo celular. La amplificación y sobreexpresión de CKS1 y SKP2 es una característica común de los tumores músculo infiltrantes, junto con la infraexpresión de p27. La amplificación de E2F3 junto con la inactivación de pRB se observó frecuentemente en nuestro grupo de estudio. La inestabilidad cromosómica es la fuerza motriz que origina los desequilibrios genómicos y que puede estar en el origen del proceso tumoral. Por ello, nos propusimos analizar muestras tumorales incluidas en parafina mediante FISH e inmunofluorescencia con el fin de estudiar las alteraciones del centrosoma y la amplificación del oncogén CCND1, respectivamente. Puesto que estas alteraciones están asociadas a la inestabilidad cromosómica numérica y estructural. Este estudio ha permitido describir el comportamiento in vivo de la amplificación de CCND1 dentro de los tumores de vejiga T1 con inestabilidad cromosómica. Se ha descrito por primera vez en la literatura la amplificación de CCND1 en forma de diminutos dobles y su posible origen en la fragmentación de las regiones de tinción homogénea. También, hemos demostrado que el agrupamiento de centrosomas supernumerarios asociado a la formación de husos mitóticos bipolares es una característica frecuente en estos tumores. El 80% de los tumores con inestabilidad cromosómica presentaron centrosomas supernumerarios. El comportamiento de los tumores uroteliales no músculo infiltrantes se caracteriza por una elevada tasa de recurrencia, lo que condiciona su seguimiento clínico. La identificación de las alteraciones moleculares críticas para la recurrencia y progresión tumoral podrían ayudar a predecir el comportamiento clínico del tumor. Por ello nos propusimos analizar la asociación entre alteraciones de la dosis génica de EGFR, CMYC, CCND1 y 9p21 con los parámetros clínicos en pacientes con cáncer de vejiga no músculo infiltrante. Este estudio se llevó a cabo en un array de 152 tumores. Nuestros resultados indican que la ganancia de EGFR asociada a polisomía del cromosoma 7 es marcador independiente de recurrencia en tumores Ta. La ganancia de 9p21 asociada a alta polisomía del cromosoma 9 y la alta ganancia de CMYC son marcadores independientes de progresión en tumores T1. El comportamiento clínico de los tumores de vejiga no músculo infiltrante comporta un control de los pacientes durante periodos de tiempo prolongados, que en algunos casos conduce a la aparición de un segundo tumor primario. Nos propusimos estudiar la incidencia de un segundo tumor primario en 231 pacientes con cáncer de vejiga no músculo infiltrante, en este grupo de pacientes se observó una incidencia de cáncer de pulmón 10,27 veces superior a la población general. Por lo que consideramos que se debería incluir una revisión anual para la detección y prevención del cáncer de pulmón, en los pacientes aquejados de cáncer de vejiga. Esta propuesta se ve reforzada por el hecho de que en el grupo de pacientes con ambas neoplasias, la muerte se produjo siempre a causa del tumor de pulmón.Bladder cancer, along with most solid tumours, is characterized by multiple numerical and structural chromosome aberrations which generate genomic imbalances. The first objective of the present study was to analyse a series of 180 urothelial bladder tumours by Comparative Genomic Hybridization in metaphase chromosomes. Our results showed a characteristic profile of genomic imbalances with gains on 1q, 8q, 11q, 16p, 17q, 18p, 19, 20q, Xq and losses on 4q, 8p, 9p, 9q, 11p, 11q, 13q 18q and Y. Gains on 5p, 6p, 10p and losses on 5q, 6q increase their frequency significantly during tumour progression. These results were subsequently confirmed by CGH array also demonstrating that muscle invasive tumours with 5p chromosome imbalances also showed amplifications at 1q, 5p, 6p and 19q affecting CKS1, SKP2, E2F3 and CCNE1 genes all of them participating in the same way of cell cycle control. We also found that amplification and over expression of CKS1 and SKP2 are characteristic of muscle invasive tumours. Amplification of E2F3 associated to pRB inactivation was frequently observed in our series. Tacking into account that chromosome instability (CIN) originates the genomic imbalances observed in bladder tumours, our second objective was to study numerical and structural chromosome instability by applying FISH, and inmunofluorescent techniques to analyse CIN index, CCND1 amplification and centrosome abnormalities in a series of 21 paraffin-embedded bladder tumours. Our results allowed describing the in vivo behaviour of CCND1 amplification in chromosomal unstable T1 bladder tumours. Our study is the first report regarding the simultaneous CCND1 amplification in DM and HSR and the possible origin of DM from HSR fragmentation. We also demonstrated that centrosome coalescence is a common event in bladder tumours being present in 80% of the most unstable tumours analyzed. The clinical behaviour of superficial bladder tumours is characterized by recurrence, which directs their clinical management. Identification of the molecular alterations critical for tumour recurrence and progression would outcome prediction and treatment selection. Our third objective was to asses the association between alterations in gene dosage and deregulation of protein expression with clinical parameters in patients with non-muscle invasive bladder carcinoma (NMIBC). To achieve this goal, the CCND1, CMYC and EGFR genes and the 9p21 locus were analyzed by fluorescent in situ hybridization and Cyclin D1, p53, p21 and p16 proteins by immunochemistry in a bladder cancer tissue microarray composed of 152 Ta and T1 tumours. Our results showed that polysomy 7 with high gain of EGFR constitute an independent prognostic factor for recurrence in NMIBC. Gain of 9p21 is an independent prognostic factor of progression in T1G3 bladder tumours. The clinical behaviour of low-risk NMIBC enables patients to be controlled for a long period, which in some cases leads to the development of a second primary tumour. We studied the second tumour incidence in a series of 231 patients with non muscle invasive bladder cancer. Our results showed that the risk of lung cancer was10.27-fold higher in patients with NMIBC as compared with the general population of Catalonia. According to these results, we consider that an annual examination for the detection and prevention of lung cancer must be included in clinical guides for patients with NMIBC. This proposal is reinforced by the finding that death in our group of patients with both tumours was always derived from lung cancer and not from bladder cancer

Effects of region of birth, educational level and age on late presentation among men who have sex with men newly diagnosed with HIV in a network of STI/HIV counselling and testing clinics in Spain

This paper analyses late presentation (LP) of HIV infection, and its determinants, among men who have sex with men (MSM) in Spain, newly diagnosed with HIV (2003-2011) in 15 sexually transmitted infection/HIV counselling and testing clinics. LP was defined as  12 months before diagnosis (12-24 months (aOR:1.4; 95% CI:1.0-2.0); > 24 months (aOR:2.2; 95% CI:1.7-3.0)). LP was less likely in MSM reporting a known HIV-infected partner as infection source or symptoms compatible with acute retroviral syndrome. 'Region of birth' interacted with 'educational level' and 'steady partner as infection source': only African and Latin-American MSM with low educational level were more likely to present late; Latin-American men attributing their infection to steady partner, but no other MSM, had LP more frequently. In Spain, HIV testing among MSM should be promoted, especially those > 34 years old and migrants with low educational level. The current recommendation that MSM be tested at least once a year is appropriate.This work has been supported with grants No. 36303/02, 36537/05 and 36794/08 from FIPSE (Fundación para la Investigación y la Prevención del Sida en España). The authors wish to thank Kathy Fitch for the English review.S

Novel Double Factor PGT strategy analyzing blastocyst stage embryos in a single NGS procedure

In families at risk from monogenic diseases affected offspring, it is fundamental the development of a suitable Double Factor Preimplantation Genetic Testing (DF-PGT) method for both single-gene analysis and chromosome complement screening. Aneuploidy is not only a major issue in advanced-maternal-age patients and balanced translocation carriers, but also the aneuploidy rate is extremely high in patients undergoing in vitro fertilization (IVF), even in young donors. To adequate NGS technology to the DF-PGT strategy four different whole genome amplification systems (Sureplex, MALBAC, and two multiple displacement amplification systems-MDA) were tested using TruSight One panel on cell lines and blastocyst trophectoderm biopsies-TE. Embryo cytogenetic status was analyzed by Nexus software. Sureplex and MALBAC DNA products were considered not suitable for PGT diagnosis due to inconsistent and poor results on Trusight one (TSO) panel. Results obtained with both MDA based methods (GEH-MDA and RG-MDA) were appropriate for direct mutation detection by TSO NGS platform. Nevertheless, RG-MDA amplification products showed better coverage and lower ADO rates than GEH-MDA. The present work also demonstrates that the same TSO sequencing data is suitable not only for the direct mutation detection, but also for the indirect mutation detection by linkage analysis of informative SNPs. The present work also demonstrates that Nexus software is competent for the detection of CNV by using with TSO sequencing data from RG-MDA products, allowing for the whole cytogenetic characterization of the embryos. In conclusion, successfully development of an innovative and promising DF-PGT strategy using TSO-NGS technology in TE biopsies, performed in-house in a single laboratory experience, has been done in the present work. Additional studies should be performed before it could be used as a diagnostic alternative in order to validate this approach for the detection of chromosomal aneuploidies

Trends in HIV testing, prevalence among first-time testers, and incidence in most-at-risk populations in Spain: the EPI-VIH Study, 2000 to 2009.

During 2000 to 2009, data on people undergoing HIV testing and on those newly diagnosed with HIV were collected in a network of 20 Spanish clinics specialising in sexually transmitted infections and/or HIV testing and counselling. The number of tests performed, overall and disaggregated by different variables, was obtained. HIV prevalence among first-time testers and HIV incidence among repeat testers were calculated. To evaluate trends, joinpoint regression models were fitted. In total, 236,939 HIV tests were performed for 165,745 individuals. Overall HIV prevalence among persons seeking HIV testing was 2.5% (95% CI: 2.4 to 2.6). Prevalence was highest in male sex workers who had sex with other men (19.0% (95% CI: 16.7 to 21.4)) and was lowest in female sex workers (0.8% (95% CI: 0.7 to 0.9)). Significant trends in prevalence were observed in men who have sex with men (MSM) (increasing) and heterosexual individuals (decreasing). The incidence analysis included 30,679 persons, 64,104 person-years (py) of follow-up and 642 seroconversions. The overall incidence rate (IR) was 1.0/100 py (95% CI: 0.9/100 to 1.1/100). Incidence was significantly higher in men and transgender females than in women (1.8/100 py (95% CI: 1.6 to 1.9), 1.2/100 py (95% CI: 0.5 to 2.8) and 0.1/100 py (95% CI: 0.09 to 0.2) respectively) and increased with age until 35–39 years. IRs in MSM and people who inject drugs were significantly greater than in heterosexual individuals (2.5/100 py (95% CI: 2.3 to 2.7), 1.6/100 py (95% CI: 1.1 to 2.2) and 0.1/100 py (95% CI: 0.09 to 0.2) respectively), and an upward trend was observed in MSM. Our results call for HIV prevention to be reinforced in MSM and transgender women in Spain.This work has been supported with grants No. 36303/02, 36537/05 and 36794/08 from FIPSE (Fundación para la Investigación y la Prevención del Sida en España). The au-thors wish to thank Kathy Fitch for the English review.S

Proteomic Analysis in Seminal Plasma of Fertile Donors and Infertile Patients with Sperm DNA Fragmentation

Seminal plasma proteomics studies could represent a new approach for the determination of molecular elements driving male infertility, resulting in a better male infertility characterization. The aim of this study is to investigate proteomic differences in seminal plasma samples from fertile and infertile individuals. For that, semen samples were selected according to semen analysis, clinical pathology, and values of sperm DNA fragmentation (alkaline and neutral Comet assay and Sperm Chromatin Dispersion test). A total of 24 seminal plasma samples classified in four groups were processed: fertile donors (FD), recurrent miscarriage patients (RM), asthenoteratozoospermic patients (ATZ), and asthenoteratozoospermic patients with varicocele (ATZ-VAR). Results obtained by 2D-differential gel electrophoresis (2D-DIGE) revealed 26 spots significantly increased in fertile donors when compared to patient groups. Also, eight spots in the ATZ group and two in the ATZ-VAR group were decreased compared to the other groups. Twenty-eight proteins were identified by mass spectrometry (MS), most of them involved in metabolic and cellular processes and with a catalytic or binding function. Protein-protein interactions through Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) tool suggest that a large part of them were associated with each other. Furthermore, most of them were associated with ubiquitin C, indicating that it could play an important regulation role, resulting in a potential male infertility biomarker