SNP in the genome-wide association study hotspot on chromosome 9p21 confers susceptibility to diabetic nephropathy in type 1 diabetes

AIMS/HYPOTHESIS: Parental type 2 diabetes mellitus increases the risk of diabetic nephropathy in offspring with type 1 diabetes mellitus. Several single nucleotide polymorphisms (SNPs) that predispose to type 2 diabetes mellitus have recently been identified. It is, however, not known whether such SNPs also confer susceptibility to diabetic nephropathy in patients with type 1 diabetes mellitus. METHODS: We genotyped nine SNPs associated with type 2 diabetes mellitus in genome-wide association studies in the Finnish population, and tested for their association with diabetic nephropathy as well as with severe retinopathy and cardiovascular disease in 2,963 patients with type 1 diabetes mellitus. Replication of significant SNPs was sought in 2,980 patients from three other cohorts. RESULTS: In the discovery cohort, rs10811661 near gene CDKN2A/B was associated with diabetic nephropathy. The association remained after robust Bonferroni correction for the total number of tests performed in this study (OR 1.33 [95% CI 1.14, 1.56], p?=?0.00045, p (36tests)?=?0.016). In the meta-analysis, the combined result for diabetic nephropathy was significant, with a fixed effects p value of 0.011 (OR 1.15 [95% CI 1.02, 1.29]). The association was particularly strong when patients with end-stage renal disease were compared with controls (OR 1.35 [95% CI 1.13, 1.60], p?=?0.00038). The same SNP was also associated with severe retinopathy (OR 1.37 [95% CI 1.10, 1.69] p?=?0.0040), but the association did not remain after Bonferroni correction (p (36tests)?=?0.14). None of the other selected SNPs was associated with nephropathy, severe retinopathy or cardiovascular disease. CONCLUSIONS/INTERPRETATION: A SNP predisposing to type 2 diabetes mellitus, rs10811661 near CDKN2A/B, is associated with diabetic nephropathy in patients with type 1 diabetes mellitus.

A comparison of two informative SNP-based strategies for typing Pseudomonas aeruginosa isolates from patients with cystic fibrosis

BACKGROUND: Molecular typing is integral for identifying Pseudomonas aeruginosa strains that may be shared between patients with cystic fibrosis (CF). We conducted a side-by-side comparison of two P. aeruginosa genotyping methods utilising informative-single nucleotide polymorphism (SNP) methods; one targeting 10 P. aeruginosa SNPs and using real-time polymerase chain reaction technology (HRM10SNP) and the other targeting 20 SNPs and based on the Sequenom MassARRAY platform (iPLEX20SNP).

METHODS: An in-silico analysis of the 20 SNPs used for the iPLEX20SNP method was initially conducted using sequence type (ST) data on the P. aeruginosa PubMLST website. A total of 506 clinical isolates collected from patients attending 11 CF centres throughout Australia were then tested by both the HRM10SNP and iPLEX20SNP assays. Type-ability and discriminatory power of the methods, as well as their ability to identify commonly shared P. aeruginosa strains, were compared.

RESULTS: The in-silico analyses showed that the 1401 STs available on the PubMLST website could be divided into 927 different 20-SNP profiles (D-value = 0.999), and that most STs of national or international importance in CF could be distinguished either individually or as belonging to closely related single- or double-locus variant groups. When applied to the 506 clinical isolates, the iPLEX20SNP provided better discrimination over the HRM10SNP method with 147 different 20-SNP and 92 different 10-SNP profiles observed, respectively. For detecting the three most commonly shared Australian P. aeruginosa strains AUST-01, AUST-02 and AUST-06, the two methods were in agreement for 80/81 (98.8%), 48/49 (97.8%) and 11/12 (91.7%) isolates, respectively.

CONCLUSIONS: The iPLEX20SNP is a superior new method for broader SNP-based MLST-style investigations of P. aeruginosa. However, because of convenience and availability, the HRM10SNP method remains better suited for clinical microbiology laboratories that only utilise real-time PCR technology and where the main interest is detection of the most highly-prevalent P. aeruginosa CF strains within Australian clinics.

Outlier SNP markers reveal fine-scale genetic structuring across European hake populations (Merluccius merluccius)

Shallow population structure is generally reported for most marine fish and explained as a consequence of high dispersal, connectivity and large population size. Targeted gene analyses and more recently genome-wide studies have challenged such view, suggesting that adaptive divergence might occur even when neutral markers provide genetic homogeneity across populations. Here, 381 SNPs located in transcribed regions were used to assess large- and fine-scale population structure in the European hake (Merluccius merluccius), a widely distributed demersal species of high priority for the European fishery. Analysis of 850 individuals from 19 locations across the entire distribution range showed evidence for several outlier loci, with significantly higher resolving power. While 299 putatively neutral SNPs confirmed the genetic break between basins (F(CT) = 0.016) and weak differentiation within basins, outlier loci revealed a dramatic divergence between Atlantic and Mediterranean populations (F(CT) range 0.275-0.705) and fine-scale significant population structure. Outlier loci separated North Sea and Northern Portugal populations from all other Atlantic samples and revealed a strong differentiation among Western, Central and Eastern Mediterranean geographical samples. Significant correlation of allele frequencies at outlier loci with seawater surface temperature and salinity supported the hypothesis that populations might be adapted to local conditions. Such evidence highlights the importance of integrating information from neutral and adaptive evolutionary patterns towards a better assessment of genetic diversity. Accordingly, the generated outlier SNP data could be used for tackling illegal practices in hake fishing and commercialization as well as to develop explicit spatial models for defining management units and stock boundaries.

SNP discovery using Next Generation Transcriptomic Sequencing in Atlantic herring (Clupea harengus)

The introduction of Next Generation Sequencing (NGS) has revolutionised population genetics, providing studies of non-model species with unprecedented genomic coverage, allowing evolutionary biologists to address questions previously far beyond the reach of available resources. Furthermore, the simple mutation model of Single Nucleotide Polymorphisms (SNPs) permits cost-effective high-throughput genotyping in thousands of individuals simultaneously. Genomic resources are scarce for the Atlantic herring (Clupea harengus), a small pelagic species that sustains high revenue fisheries. This paper details the development of 578 SNPs using a combined NGS and high-throughput genotyping approach. Eight individuals covering the species distribution in the eastern Atlantic were bar-coded and multiplexed into a single cDNA library and sequenced using the 454 GS FLX platform. SNP discovery was performed by de novo sequence clustering and contig assembly, followed by the mapping of reads against consensus contig sequences. Selection of candidate SNPs for genotyping was conducted using an in silico approach. SNP validation and genotyping were performed simultaneously using an Illumina 1,536 GoldenGate assay. Although the conversion rate of candidate SNPs in the genotyping assay cannot be predicted in advance, this approach has the potential to maximise cost and time efficiencies by avoiding expensive and time-consuming laboratory stages of SNP validation. Additionally, the in silico approach leads to lower ascertainment bias in the resulting SNP panel as marker selection is based only on the ability to design primers and the predicted presence of intron-exon boundaries. Consequently SNPs with a wider spectrum of minor allele frequencies (MAFs) will be genotyped in the final panel. The genomic resources presented here represent a valuable multi-purpose resource for developing informative marker panels for population discrimination, microarray development and for population genomic studies in the wild.

Novel tools for conservation genomics: comparing two high-throughput approaches for SNP discovery in the transcriptome of the European hake

The growing accessibility to genomic resources using next-generation sequencing (NGS) technologies has revolutionized the application of molecular genetic tools to ecology and evolutionary studies in non-model organisms. Here we present the case study of the European hake (Merluccius merluccius), one of the most important demersal resources of European fisheries. Two sequencing platforms, the Roche 454 FLX (454) and the Illumina Genome Analyzer (GAII), were used for Single Nucleotide Polymorphisms (SNPs) discovery in the hake muscle transcriptome. De novo transcriptome assembly into unique contigs, annotation, and in silico SNP detection were carried out in parallel for 454 and GAII sequence data. High-throughput genotyping using the Illumina GoldenGate assay was performed for validating 1,536 putative SNPs. Validation results were analysed to compare the performances of 454 and GAII methods and to evaluate the role of several variables (e.g. sequencing depth, intron-exon structure, sequence quality and annotation). Despite well-known differences in sequence length and throughput, the two approaches showed similar assay conversion rates (approximately 43%) and percentages of polymorphic loci (67.5% and 63.3% for GAII and 454, respectively). Both NGS platforms therefore demonstrated to be suitable for large scale identification of SNPs in transcribed regions of non-model species, although the lack of a reference genome profoundly affects the genotyping success rate. The overall efficiency, however, can be improved using strict quality and filtering criteria for SNP selection (sequence quality, intron-exon structure, target region score).

Immunochip SNP array identifies novel genetic variants conferring susceptibility to candidaemia.

Candidaemia is the fourth most common cause of bloodstream infection, with a high mortality rate of up to 40%. Identification of host genetic factors that confer susceptibility to candidaemia may aid in designing adjunctive immunotherapeutic strategies. Here we hypothesize that variation in immune genes may predispose to candidaemia. We analyse 118,989 single-nucleotide polymorphisms (SNPs) across 186 loci known to be associated with immune-mediated diseases in the largest candidaemia cohort to date of 217 patients of European ancestry and a group of 11,920 controls. We validate the significant associations by comparison with a disease-matched control group. We observe significant association between candidaemia and SNPs in the CD58 (P = 1.97 × 10(-11); odds ratio (OR) = 4.68), LCE4A-C1orf68 (P = 1.98 × 10(-10); OR = 4.25) and TAGAP (P = 1.84 × 10(-8); OR = 2.96) loci. Individuals carrying two or more risk alleles have an increased risk for candidaemia of 19.4-fold compared with individuals carrying no risk allele. We identify three novel genetic risk factors for candidaemia, which we subsequently validate for their role in antifungal host defence.

Situación de los stocks de peces demersales en el otoño de 1990 : Crucero BIC SNP-l 9005-06 (19 de mayo - 08 de junio 1990) (Huarmey - Puerto Pizarro)

Analiza los resultados del crucero de evaluación de peces demersales con especial referencia a la merluza, para la plataforma continental comprendida entre la frontera con el Ecuador y 10º S, a profundidades entre 20 y 200 bz, totalizando 13542,27 millas náuticas cuadradas.

Evaluación de los recursos pelágicos: anchoveta, sardina, jurel y caballa en primavera 1989 : Crucero de evaluacion acustica BIC/SNP-1 8911-12 (Chicama - Punta Doña Maria)

Describe los principales resultados obtenidos en el Crucero evaluación de recursos pelágicos, efectuado a bordo del BIC/SNP-1 del 14 de noviembre de 1989, entre Chicama y Punta Doña María, constituyendo la segunda prospección planificada dentro del marco del proyecto Evaluación de los recursos: anchoveta, sardina, jurel y caballa, auspiciado por el Instituto del mar del Perú y la Comunidad económica europea.

Biomasa desovante de la anchoveta y condiciones oceanográficas. Crucero 9408- 10 BIC SNP-1 y bolicheras

Describe las actividades realizadas en el crucero de evaluación de biomasa desovante de anchoveta y sardina, realizadas desde el 11 de agosto al 4 de octubre de 1994, con el objetivo de evaluar a las poblaciones desovantes de la anchoveta y sardina del litoral, para lo cual exploró el área comprendida entre los 10°00 'S y 17°30 'S, llegando hasta las 102 mm de la costa. Los registros acústicos mostraron que la distribución de la anchoveta fue amplia, desde las 02 hasta las 102 millas de la costa, formando grandes áreas de concentración, principalmente frente a Chimbote, Salaverry, Chicama y Pimentel y en menor densidad entre Huarmey y Pisco. La sardina en cambio se presentó en pequeñas áreas dispersas entre Pimentel y Paita, desde las 15mn hasta las 102 millas de distancia de la costa. Las concentraciones y distribución de la especie no permitieron realizar el estimado de biomasa desovante. La Biomasa desovante estimada para ala anchoveta mediante el Método de Producción de huevos es de 6,9 millones de toneladas, con una producción de 15,12 E + 13 huevos por día. La temperatura superficial del mar fluctuó entre 15.0 y 20,4° C. En general, en todo el litoral se observó una tendencia homogénea, con algunos incrementos de carácter temporal al inicio de la primavera.

Aspectos biológicos - pesqueros de los recursos pelágicos. Crucero de evaluación de recursos pelágicos 9502-04 BIC SNP-1 (13 febrero - 05 abril, 1995)

Describe los niveles de abundancia , distribución, concentración y características biológicas de las poblaciones de la anchoveta y sardina a principios de 1995, así como conocer las características del ambiente y su influencia sobre el reclutamiento. La información proviene de las capturas y muestreos biológicos de 129 lances de comprobación ejecutados durante el crucero. La composición por especies estuvo conformada principalmente por anchoveta (Engraulis ringens), sardina (Sardinops sagax sagax), jurel (Trachurus picturatus murphyi) y caballa (Scomber japonicus peruanus). Otras especies fueron el bagre con faja (Galeichthys peruvianus), falso volador (Prionotus stephanophrys), múnida (Pleuroncodes monodon), etc.

Condiciones oceanográficas del mar peruano durante el crucero de evaluación de recursos pelágicos 9502-04 BIC/SNP-1 (13 febrero - 05 abril, 1995)

Presenta los resultados de las condiciones físicas y químicas (contenido de oxígeno disuelto y clorofila "a") observadas en superficie y capas subsuperficiales del mar y se dan perspectivas del comportamiento de ambiente para los próximos meses. Las características del ambiente indicaron un calentamiento de magnitud moderada al sur del Callao y condiciones cuasi-normales al norte del Callao que confirmaron la continuación del proceso de normalización del mar peruano iniciado en febrero de 1995.

Fitoplancton durante el crucero de evaluación de recursos pelágicos (13 febrero - 05 abril, 1995) BIC SNP-1

Describe las muestras de fitoplancton en 127 estaciones de Tacna a Tumbes. Paralelamente en los perfiles hidrográficos se colectaron muestras de agua con botellas Niskin. Se determinó un total de 143 especies de fitoplancton: 86 diatomeas, 44 dinoflagelados, 7 cocolitofóridos, 6 fitoflagelados y 2 silicoflagelados. Las concentraciones máximas de fitoplancton, mayores de 10.000 cel/50 mL, a 10 m de profundidad se encontraron en la franja costera, predominando diatomeas de alta tasa de reproducción, las cuales corresponden a una comunidad en la primera fase de la sucesión fitoplanctónica.