| 1. |
|
|
| 2. |
|
|
| 3. |
|
|
| 4. |
- van Rheenen, Wouter, et al.
(author)
-
Genome-wide association analyses identify new risk variants and the genetic architecture of amyotrophic lateral sclerosis
- 2016
-
In: Nature Genetics. - : Springer Science and Business Media LLC. - 1061-4036 .- 1546-1718. ; 48:9, s. 1043-1048
-
Journal article (peer-reviewed)abstract
- To elucidate the genetic architecture of amyotrophic lateral sclerosis (ALS) and find associated loci, we assembled a custom imputation reference panel from whole-genome-sequenced patients with ALS and matched controls (n = 1,861). Through imputation and mixed-model association analysis in 12,577 cases and 23,475 controls, combined with 2,579 cases and 2,767 controls in an independent replication cohort, we fine-mapped a new risk locus on chromosome 21 and identified C21orf2 as a gene associated with ALS risk. In addition, we identified MOBP and SCFD1 as new associated risk loci. We established evidence of ALS being a complex genetic trait with a polygenic architecture. Furthermore, we estimated the SNP-based heritability at 8.5%, with a distinct and important role for low-frequency variants (frequency 1-10%). This study motivates the interrogation of larger samples with full genome coverage to identify rare causal variants that underpin ALS risk.
|
|
| 5. |
- Blauw, Hylke M, et al.
(author)
-
A large genome scan for rare CNVs in amyotrophic lateral sclerosis
- 2010
-
In: Human Molecular Genetics. - : Oxford Journals. - 0964-6906 .- 1460-2083. ; 19:20, s. 4091-4099
-
Journal article (peer-reviewed)abstract
- Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease selectively affecting motor neurons in the brain and spinal cord. Recent genome-wide association studies (GWASs) have identified several common variants which increase disease susceptibility. In contrast, rare copy-number variants (CNVs), which have been associated with several neuropsychiatric traits, have not been studied for ALS in well-powered study populations. To examine the role of rare CNVs in ALS susceptibility, we conducted a CNV association study including over 19,000 individuals. In a genome-wide screen of 1875 cases and 8731 controls, we did not find evidence for a difference in global CNV burden between cases and controls. In our association analyses, we identified two loci that met our criteria for follow-up: the DPP6 locus (OR = 3.59, P = 6.6 × 10(-3)), which has already been implicated in ALS pathogenesis, and the 15q11.2 locus, containing NIPA1 (OR = 12.46, P = 9.3 × 10(-5)), the gene causing hereditary spastic paraparesis type 6 (HSP 6). We tested these loci in a replication cohort of 2559 cases and 5887 controls. Again, results were suggestive of association, but did not meet our criteria for independent replication: DPP6 locus: OR = 1.92, P = 0.097, pooled results: OR = 2.64, P = 1.4 × 10(-3); NIPA1: OR = 3.23, P = 0.041, pooled results: OR = 6.20, P = 2.2 × 10(-5)). Our results highlight DPP6 and NIPA1 as candidates for more in-depth studies. Unlike other complex neurological and psychiatric traits, rare CNVs with high effect size do not play a major role in ALS pathogenesis.
|
|
| 6. |
- Lindblad-Toh, Kerstin, et al.
(author)
-
Genome sequence, comparative analysis and haplotype structure of the domestic dog.
- 2005
-
In: Nature. - : Springer Science and Business Media LLC. - 1476-4687 .- 0028-0836. ; 438:7069, s. 803-19
-
Journal article (peer-reviewed)abstract
- Here we report a high-quality draft genome sequence of the domestic dog (Canis familiaris), together with a dense map of single nucleotide polymorphisms (SNPs) across breeds. The dog is of particular interest because it provides important evolutionary information and because existing breeds show great phenotypic diversity for morphological, physiological and behavioural traits. We use sequence comparison with the primate and rodent lineages to shed light on the structure and evolution of genomes and genes. Notably, the majority of the most highly conserved non-coding sequences in mammalian genomes are clustered near a small subset of genes with important roles in development. Analysis of SNPs reveals long-range haplotypes across the entire dog genome, and defines the nature of genetic diversity within and across breeds. The current SNP map now makes it possible for genome-wide association studies to identify genes responsible for diseases and traits, with important consequences for human and companion animal health.
|
|
| 7. |
- Carneiro, Ana P. B., et al.
(author)
-
A framework for mapping the distribution of seabirds by integrating tracking, demography and phenology
- 2020
-
In: Journal of Applied Ecology. - : Wiley. - 0021-8901 .- 1365-2664. ; 57:3, s. 514-525
-
Journal article (peer-reviewed)abstract
- The identification of geographic areas where the densities of animals are highest across their annual cycles is a crucial step in conservation planning. In marine environments, however, it can be particularly difficult to map the distribution of species, and the methods used are usually biased towards adults, neglecting the distribution of other life-history stages even though they can represent a substantial proportion of the total population. Here we develop a methodological framework for estimating population-level density distributions of seabirds, integrating tracking data across the main life-history stages (adult breeders and non-breeders, juveniles and immatures). We incorporate demographic information (adult and juvenile/immature survival, breeding frequency and success, age at first breeding) and phenological data (average timing of breeding and migration) to weight distribution maps according to the proportion of the population represented by each life-history stage. We demonstrate the utility of this framework by applying it to 22 species of albatrosses and petrels that are of conservation concern due to interactions with fisheries. Because juveniles, immatures and non-breeding adults account for 47%-81% of all individuals of the populations analysed, ignoring the distributions of birds in these stages leads to biased estimates of overlap with threats, and may misdirect management and conservation efforts. Population-level distribution maps using only adult distributions underestimated exposure to longline fishing effort by 18%-42%, compared with overlap scores based on data from all life-history stages. Synthesis and applications. Our framework synthesizes and improves on previous approaches to estimate seabird densities at sea, is applicable for data-poor situations, and provides a standard and repeatable method that can be easily updated as new tracking and demographic data become available. We provide scripts in the R language and a Shiny app to facilitate future applications of our approach. We recommend that where sufficient tracking data are available, this framework be used to assess overlap of seabirds with at-sea threats such as overharvesting, fisheries bycatch, shipping, offshore industry and pollutants. Based on such an analysis, conservation interventions could be directed towards areas where they have the greatest impact on populations.
|
|
| 8. |
- Evander, Mikael, et al.
(author)
-
Acoustic Differential Extraction - ultrasonic DNA-extraction from sexual assault evidence
- 2007
-
In: International Congress on Ultrasonics. ; 1, s. 151-151
-
Conference paper (peer-reviewed)abstract
- Isolation of the male and female DNA is one of the most important steps in obtaining the DNA profile of the perpetrator in sexual assault cases. The sample is obtained by taking a vaginal swab containing both epithelial cells from the female and sperm cells from the male from the victim. The purity of the extracted male fraction decides whether or not a single-source DNA profile of the suspect will be obtained or not. The existing techniques are have poor separation efficiency, time-consuming, labour-intensive and are neither easily automated nor integrated with further analysis steps. A novel method of DNA extraction based on ultrasonic trapping, Acoustic Differential Extraction, has been developed. A microfluidic device using laminar flow valving and miniature PZT transducers retains the sperm cells while mobilizing the female fraction into a separate outlet. The device was evaluated using a mock sexual assault sample and the separated fractions were analyzed using quantitative PCR and STR-profiling. A 16-fold enrichment of the male fraction, making an originally hard-to-detect-male DNA profile readily profiled, has been demonstrated. The STR-profiling of the male and female fractions showed a male fraction purity of up to 99 % making it possible to obtain a single-source DNA-profile of the suspect. The microfluidic format of the device makes it possible to downscale the sample time from 4-8 hours to 10 minutes. It is also possible to integrate this method with further downstream analysis steps necessarey for the full forensic DNA analysis.
|
|
| 9. |
- Evander, Mikael, et al.
(author)
-
Using Acoustic Differential Extraction to enhance analysis of sexual assualt evidence on a valveless glass microdevice
- 2006
-
In: Proceedings of µTAS 2006 Conference. ; 2, s. 1055-1057
-
Conference paper (peer-reviewed)abstract
- The isolation of male and female DNA is an important step in the analysis of sexual assault evidence. A vaginal swab with female epithelial cells and male sperm cells is obtained from the female, and it is vital to separate the male and female fractions in order to obtain a single-source DNA profile of the suspect. In the case of a low abundance of sperm cells, it is very important that no cells are lost in the isolation step. The conventional isolation method used in the forensic DNA laboratories, differential extraction, is a time-consuming step, requiring up to 24 hours. It is neither highly amenable to automation, nor can it be easily integrated with other steps of the analysis. Therefore, a novel method of performing the isolation of male and female fractions of biological material from sexual assault evidence has been developed, termed acoustic differential extraction (ADE). After selectively lysing the female epithelial cells while keeping the sperm cells intact, the sample, now containing sperm cells and female cell lysate, is infused in a 900 μm wide and 70 μm deep microfabricated glass channel with miniature piezoelectric transducers mounted at the bottom of the channel, as shown in Figure 1. Upon activation of the ultrasound, the sperm cells will be trapped in a standing wave1 while free DNA will not be retained. The sperm cells, levitated in the 3D fluidic space above the transducer, can be washed with buffer and the unretained biological material directed to an output reservoir. Using laminar flow valving2, the sperm cells can be released and directed into a separate output reservoir in anticipation of DNA analysis, see Figure 2. With the purpose of evaluating the ADE microdevice for the collection of the two output fractions (male and female), preliminary work used a mock sexual assault sample created with polystyrene microparticles as sperm cells and Evan's Blue dye as female cell lysate. The particles were trapped over the transducer and the dye was directed to the female outlet reservoir as shown in Figure 3. After washing the particles, the ultrasound was deactivated and the flow redirected in order to collect the particles in the male outlet reservoir. A biological sample consisting of sperm cells and lysed female epithelial cells was subsequently tested by infusion into the device for five minutes while trapping the sperm cells over the transducer (Figure 4). The trapped sperm cells were washed with PBS for five minutes, then released and collected for analysis off-chip. DNA from the isolated cells was extracted with a commercial DNA extraction kit and analyzed with a duplex quantitative PCR assay3 to show the sample purity. An example of the qPCR data obtained is provided in Table 1. The results show that a highly-enriched sperm cell fraction can be obtained with the ADE technique. It is reasonable to expect that this technique can be integrated with on-chip downstream sample processing, e.g. DNA extraction and amplification. This would greatly diminish the analysis time from 24 hours to approximately 60 minutes. The time savings, in combination with the possibility to create a fully automated system, gives the ADE technique the potential to significantly alter the means by which sexual assault evidence is processed in crime laboratories today.
|
|
| 10. |
- Kenna, Kevin P., et al.
(author)
-
NEK1 variants confer susceptibility to amyotrophic lateral sclerosis
- 2016
-
In: Nature Genetics. - : Springer Science and Business Media LLC. - 1061-4036 .- 1546-1718. ; 48:9, s. 1037-1042
-
Journal article (peer-reviewed)abstract
- To identify genetic factors contributing to amyotrophic lateral sclerosis (ALS), we conducted whole-exome analyses of 1,022 index familial ALS (FALS) cases and 7,315 controls. In a new screening strategy, we performed gene-burden analyses trained with established ALS genes and identified a significant association between loss-of-function (LOF) NEK1 variants and FALS risk. Independently, autozygosity mapping for an isolated community in the Netherlands identified a NEK1 p.Arg261 His variant as a candidate risk factor. Replication analyses of sporadic ALS (SALS) cases and independent control cohorts confirmed significant disease association for both p.Arg261 His (10,589 samples analyzed) and NEK1 LOF variants (3,362 samples analyzed). In total, we observed NEK1 risk variants in nearly 3% of ALS cases. NEK1 has been linked to several cellular functions, including cilia formation, DNA-damage response, microtubule stability, neuronal morphology and axonal polarity. Our results provide new and important insights into ALS etiopathogenesis and genetic etiology.
|
|
| 11. |
- Lloyd-Price, Jason, et al.
(author)
-
Multi-omics of the gut microbial ecosystem in inflammatory bowel diseases
- 2019
-
In: Nature. - : Nature Publishing Group. - 0028-0836 .- 1476-4687. ; 569:7758, s. 655-661
-
Journal article (peer-reviewed)abstract
- Inflammatory bowel diseases, which include Crohn's disease and ulcerative colitis, affect several million individuals worldwide. Crohn's disease and ulcerative colitis are complex diseases that are heterogeneous at the clinical, immunological, molecular, genetic, and microbial levels. Individual contributing factors have been the focus of extensive research. As part of the Integrative Human Microbiome Project (HMP2 or iHMP), we followed 132 subjects for one year each to generate integrated longitudinal molecular profiles of host and microbial activity during disease (up to 24 time points each; in total 2,965 stool, biopsy, and blood specimens). Here we present the results, which provide a comprehensive view of functional dysbiosis in the gut microbiome during inflammatory bowel disease activity. We demonstrate a characteristic increase in facultative anaerobes at the expense of obligate anaerobes, as well as molecular disruptions in microbial transcription (for example, among clostridia), metabolite pools (acylcarnitines, bile acids, and short-chain fatty acids), and levels of antibodies in host serum. Periods of disease activity were also marked by increases in temporal variability, with characteristic taxonomic, functional, and biochemical shifts. Finally, integrative analysis identified microbial, biochemical, and host factors central to this dysregulation. The study's infrastructure resources, results, and data, which are available through the Inflammatory Bowel Disease Multi'omics Database (http://ibdmdb.org), provide the most comprehensive description to date of host and microbial activities in inflammatory bowel diseases.
|
|
| 12. |
- Marriott, Heather, et al.
(author)
-
Mutations in the tail and rod domains of the neurofilament heavy-chain gene increase the risk of ALS
- 2024
-
In: Annals of Clinical and Translational Neurology. - : John Wiley & Sons. - 2328-9503.
-
Journal article (peer-reviewed)abstract
- Objective: Neurofilament heavy-chain gene (NEFH) variants are associated with multiple neurodegenerative diseases, however, their relationship with ALS has not been robustly explored. Still, NEFH is commonly included in genetic screening panels worldwide. We therefore aimed to determine if NEFH variants modify ALS risk.Methods: Genetic data of 11,130 people with ALS and 7,416 controls from the literature and Project MinE were analysed. We performed meta-analyses of published case–control studies reporting NEFH variants, and variant analysis of NEFH in Project MinE whole-genome sequencing data.Results: Fixed-effects meta-analysis found that rare (MAF <1%) missense variants in the tail domain of NEFH increase ALS risk (OR 4.55, 95% CI 2.13–9.71, p < 0.0001). In Project MinE, ultrarare NEFH variants increased ALS risk (OR 1.37 95% CI 1.14–1.63, p = 0.0007), with rod domain variants (mostly intronic) appearing to drive the association (OR 1.45 95% CI 1.18–1.77, pMadsen–Browning = 0.0007, pSKAT-O = 0.003). While in the tail domain, ultrarare (MAF <0.1%) pathogenic missense variants were also associated with higher risk of ALS (OR 1.94, 95% CI 0.86–4.37, pMadsen–Browning = 0.039), supporting the meta-analysis results. Finally, several tail in-frame deletions were also found to affect disease risk, however, both protective and pathogenic deletions were found in this domain, highlighting an intricated architecture that requires further investigation.Interpretation: We showed that NEFH tail missense and in-frame deletion variants, and intronic rod variants are risk factors for ALS. However, they are not variants of large effect, and their functional impact needs to be clarified in further studies. Therefore, their inclusion in routine genetic screening panels should be reconsidered.
|
|
| 13. |
- Nicolas, Aude, et al.
(author)
-
Genome-wide Analyses Identify KIF5A as a Novel ALS Gene
- 2018
-
In: Neuron. - : Cell Press. - 0896-6273 .- 1097-4199. ; 97:6, s. 1268-1283.e6
-
Journal article (peer-reviewed)abstract
- To identify novel genes associated with ALS, we undertook two lines of investigation. We carried out a genome-wide association study comparing 20,806 ALS cases and 59,804 controls. Independently, we performed a rare variant burden analysis comparing 1,138 index familial ALS cases and 19,494 controls. Through both approaches, we identified kinesin family member 5A (KIF5A) as a novel gene associated with ALS. Interestingly, mutations predominantly in the N-terminal motor domain of KIF5A are causative for two neurodegenerative diseases: hereditary spastic paraplegia (SPG10) and Charcot-Marie-Tooth type 2 (CMT2). In contrast, ALS-associated mutations are primarily located at the C-terminal cargo-binding tail domain and patients harboring loss-of-function mutations displayed an extended survival relative to typical ALS cases. Taken together, these results broaden the phenotype spectrum resulting from mutations in KIF5A and strengthen the role of cytoskeletal defects in the pathogenesis of ALS.
|
|
| 14. |
- Norris, Jessica Voorhees, et al.
(author)
-
Acoustic differential extraction for forensic analysis of sexual assault evidence.
- 2009
-
In: Analytical Chemistry. - : American Chemical Society (ACS). - 1520-6882 .- 0003-2700. ; 81:15, s. 6089-6095
-
Journal article (peer-reviewed)abstract
- Forensic DNA analysis of samples obtained from sexual assault evidence relies on separation of male and female components of the recovered genetic material. The conventional separation method used by crime laboratories, differential extraction (DE), is one of the most time-consuming sample preparation steps, requires extensive sample handling, is difficult to automate, and often results in inefficient separation of female DNA from the male sample components. To circumvent conventional DE, acoustic differential extraction (ADE) analysis was developed on a microfluidic device. The ADE method relies on acoustic trapping of sperm cells in the presence of epithelial cell lysate (which is unretained), and laminar flow valving to direct the male and female fractions to separate outlets. Following the separation of sperm from epithelial cell lysate, DNA extraction, quantitation, amplification, and separation were performed using conventional laboratory methods. The results show that highly purified male and female fractions can be obtained with the ADE microdevice from mock sexual assault samples in 14 min. ADE analysis provides the potential to significantly alter the means by which sexual assault evidence is processed in crime laboratories.
|
|
| 15. |
|
|
| 16. |
|
|
| 17. |
- van Es, Michael A, et al.
(author)
-
Angiogenin variants in Parkinson disease and amyotrophic lateral sclerosis
- 2011
-
In: Annals of Neurology. - : Wiley-Blackwell. - 0364-5134 .- 1531-8249. ; 70:6, s. 964-973
-
Journal article (peer-reviewed)abstract
- OBJECTIVE: Several studies have suggested an increased frequency of variants in the gene encoding angiogenin (ANG) in patients with amyotrophic lateral sclerosis (ALS). Interestingly, a few ALS patients carrying ANG variants also showed signs of Parkinson disease (PD). Furthermore, relatives of ALS patients have an increased risk to develop PD, and the prevalence of concomitant motor neuron disease in PD is higher than expected based on chance occurrence. We therefore investigated whether ANG variants could predispose to both ALS and PD.METHODS: We reviewed all previous studies on ANG in ALS and performed sequence experiments on additional samples, which allowed us to analyze data from 6,471 ALS patients and 7,668 controls from 15 centers (13 from Europe and 2 from the USA). We sequenced DNA samples from 3,146 PD patients from 6 centers (5 from Europe and 1 from the USA). Statistical analysis was performed using the variable threshold test, and the Mantel-Haenszel procedure was used to estimate odds ratios.RESULTS: Analysis of sequence data from 17,258 individuals demonstrated a significantly higher frequency of ANG variants in both ALS and PD patients compared to control subjects (p = 9.3 × 10(-6) for ALS and p = 4.3 × 10(-5) for PD). The odds ratio for any ANG variant in patients versus controls was 9.2 for ALS and 6.7 for PD.INTERPRETATION: The data from this multicenter study demonstrate that there is a strong association between PD, ALS, and ANG variants. ANG is a genetic link between ALS and PD.
|
|
| 18. |
- van Es, Michael A, et al.
(author)
-
Genome-wide association study identifies 19p13.3 (UNC13A) and 9p21.2 as susceptibility loci for sporadic amyotrophic lateral sclerosis
- 2009
-
In: Nature genetics. - : Springer Science and Business Media LLC. - 1546-1718 .- 1061-4036. ; 41:10, s. 1083-1087
-
Journal article (peer-reviewed)abstract
- We conducted a genome-wide association study among 2,323 individuals with sporadic amyotrophic lateral sclerosis (ALS) and 9,013 control subjects and evaluated all SNPs with P < 1.0 x 10(-4) in a second, independent cohort of 2,532 affected individuals and 5,940 controls. Analysis of the genome-wide data revealed genome-wide significance for one SNP, rs12608932, with P = 1.30 x 10(-9). This SNP showed robust replication in the second cohort (P = 1.86 x 10(-6)), and a combined analysis over the two stages yielded P = 2.53 x 10(-14). The rs12608932 SNP is located at 19p13.3 and maps to a haplotype block within the boundaries of UNC13A, which regulates the release of neurotransmitters such as glutamate at neuromuscular synapses. Follow-up of additional SNPs showed genome-wide significance for two further SNPs (rs2814707, with P = 7.45 x 10(-9), and rs3849942, with P = 1.01 x 10(-8)) in the combined analysis of both stages. These SNPs are located at chromosome 9p21.2, in a linkage region for familial ALS with frontotemporal dementia found previously in several large pedigrees.
|
|
| 19. |
- Xu, Kerui, et al.
(author)
-
Isolation of a Low Number of Sperm Cells from Female DNA in a Glass-PDMS-Glass Microchip via Bead-Assisted Acoustic Differential Extraction
- 2019
-
In: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 91:3, s. 2186-2191
-
Journal article (peer-reviewed)abstract
- We report an improved separation method for the isolation of sperm cells from dilute, "large volume" samples containing female DNA using bead-assisted acoustic trapping. In an enclosed glass-PDMS-glass (GPG) resonator, we exploit a three-layer microfluidic architecture to generate "trapping nodes" in ultrasonic standing waves. We investigate the dependence of trapping efficiency on particle concentration for both sperm cells and polymeric beads. After determination of the critical concentration of polymeric beads required to seed the trapping event, sperm cells in dilute solution are trapped as a result of the enhanced secondary radiation force (SRF). Sperm-cell-containing samples with volumes up to 300 μL and cell concentrations as low as ∼10 cells/μL are amenable to effective trapping in the presence of an abundance of female DNA in solution. Complete processing of samples is accomplished with separation of the female and male fractions within 15 min. We demonstrate that the collected fractions are amenable to subsequent DNA extraction, short tandem repeat PCR, and the generation of STR profiles for the isolated sperm cells.
|
|
