Genotype imputation using the Positional Burrows Wheeler Transform.

Genotype imputation is the process of predicting unobserved genotypes in a sample of individuals using a reference panel of haplotypes. In the last 10 years reference panels have increased in size by more than 100 fold. Increasing reference panel size improves accuracy of markers with low minor allele frequencies but poses ever increasing computational challenges for imputation methods. Here we present IMPUTE5, a genotype imputation method that can scale to reference panels with millions of samples. This method continues to refine the observation made in the IMPUTE2 method, that accuracy is optimized via use of a custom subset of haplotypes when imputing each individual. It achieves fast, accurate, and memory-efficient imputation by selecting haplotypes using the Positional Burrows Wheeler Transform (PBWT). By using the PBWT data structure at genotyped markers, IMPUTE5 identifies locally best matching haplotypes and long identical by state segments. The method then uses the selected haplotypes as conditioning states within the IMPUTE model. Using the HRC reference panel, which has ∼65,000 haplotypes, we show that IMPUTE5 is up to 30x faster than MINIMAC4 and up to 3x faster than BEAGLE5.1, and uses less memory than both these methods. Using simulated reference panels we show that IMPUTE5 scales sub-linearly with reference panel size. For example, keeping the number of imputed markers constant, increasing the reference panel size from 10,000 to 1 million haplotypes requires less than twice the computation time. As the reference panel increases in size IMPUTE5 is able to utilize a smaller number of reference haplotypes, thus reducing computational cost

Phasing for medical sequencing using rare variants and large haplotype reference panels.

Motivation: There is growing recognition that estimating haplotypes from high coverage sequencing of single samples in clinical settings is an important problem. At the same time very large datasets consisting of tens and hundreds of thousands of high-coverage sequenced samples will soon be available. We describe a method that takes advantage of these huge human genetic variation resources and rare variant sharing patterns to estimate haplotypes on single sequenced samples. Sharing rare variants between two individuals is more likely to arise from a recent common ancestor and, hence, also more likely to indicate similar shared haplotypes over a substantial flanking region of sequence.Results: Our method exploits this idea to select a small set of highly informative copying states within a Hidden Markov Model (HMM) phasing algorithm. Using rare variants in this way allows us to avoid iterative MCMC methods to infer haplotypes. Compared to other approaches that do not explicitly use rare variants we obtain significant gains in phasing accuracy, less variation over phasing runs and improvements in speed. For example, using a reference panel of 7420 haplotypes from the UK10K project, we are able to reduce switch error rates by up to 50% when phasing samples sequenced at high-coverage. In addition, a single step rephasing of the UK10K panel, using rare variant information, has a downstream impact on phasing performance. These results represent a proof of concept that rare variant sharing patterns can be utilized to phase large high-coverage sequencing studies such as the 100 000 Genomes Project dataset.</br

The non-indigenous Paranthura japonica Richardson, 1909 in the Mediterranean Sea: travelling with shellfish?

An anthurid isopod new to the Mediterranean Sea has recently been observed in samples from three localities of the Italian coast: the Lagoon of Venice (North Adriatic Sea), La Spezia (Ligurian Sea) and Olbia (Sardinia, Tyrrhenian Sea). The specimens collected showed strong affinity to a species originally described from the NW Pacific Ocean: Paranthura japonica Richardson, 1909. The comparison with specimens collected from the Bay of Arcachon (Atlantic coast of France), where P. japonica had been recently reported as non-indigenous, confirmed the identity of the species. This paper reports the most relevant morphological details of the Italian specimens, data on the current distribution of the species and a discussion on the pathways responsible for its introduction. The available data suggest that the presence of this Pacific isopod in several regions of coastal Europe might be due to a series of aquaculture-mediated introduction events that occurred during the last decades of the 1900s. Since then, established populations of P. japonica, probably misidentified, remained unnoticed for a long time

Mitogen-activated protein kinase kinase 5 regulates proliferation and biosynthetic processes in procyclic forms of Trypanosoma brucei

The pathogenic protozoan T. brucei alternates into distinct developmental stages in the mammalian and insect hosts. The mitogen-activated protein kinase (MAPK) signaling pathways transduce extracellular stimuli into a range of cellular responses, which ultimately lead to the adaptation to the external environment. Here, we combined a loss of function approach with stable isotope labeling with amino acids in cell culture (SILAC)-based mass spectrometry (MS) to investigate the role of the mitogen-activated protein kinase kinase 5 (MKK5) in T. brucei. The silencing of MKK5 significantly decreased the proliferation of procyclic forms of T. brucei. To shed light on the molecular alterations associated with this phenotype, we measured the total proteome and phosphoproteome of cells silenced for MKK5. In the total proteome, we observed a general decrease in proteins related to ribosome and translation as well as down-regulation of several components of the fatty acids biosynthesis pathway. In addition, we observed alterations in the protein levels and phosphorylation of key metabolic enzymes, which point toward a suppression of the oxidative metabolism. Taken together, our findings show that the silencing of MKK5 alters cell growth, energy metabolism, protein and fatty acids biosynthesis in procyclic T. brucei

Unit cell of graphene on Ru(0001): a 25 x 25 supercell with 1250 carbon atoms

The structure of a single layer of graphene on Ru(0001) has been studied using surface x-ray diffraction. A surprising superstructure has been determined, whereby 25 x 25 graphene unit cells lie on 23 x 23 unit cells of Ru. Each supercell contains 2 x 2 crystallographically inequivalent subcells caused by corrugation. Strong intensity oscillations in the superstructure rods demonstrate that the Ru substrate is also significantly corrugated down to several monolayers, and that the bonding between graphene and Ru is strong and cannot be caused by van der Waals bonds. Charge transfer from the Ru substrate to the graphene expands and weakens the C-C bonds, which helps accommodate the in-plane tensile stress. The elucidation of this superstructure provides important information in the potential application of graphene as a template for nanocluster arrays.Comment: 9 pages, 3 figures, paper submitted to peer reviewed journa

A Flexible and Accurate Genotype Imputation Method for the Next Generation of Genome-Wide Association Studies

Genotype imputation methods are now being widely used in the analysis of genome-wide association studies. Most imputation analyses to date have used the HapMap as a reference dataset, but new reference panels (such as controls genotyped on multiple SNP chips and densely typed samples from the 1,000 Genomes Project) will soon allow a broader range of SNPs to be imputed with higher accuracy, thereby increasing power. We describe a genotype imputation method (IMPUTE version 2) that is designed to address the challenges presented by these new datasets. The main innovation of our approach is a flexible modelling framework that increases accuracy and combines information across multiple reference panels while remaining computationally feasible. We find that IMPUTE v2 attains higher accuracy than other methods when the HapMap provides the sole reference panel, but that the size of the panel constrains the improvements that can be made. We also find that imputation accuracy can be greatly enhanced by expanding the reference panel to contain thousands of chromosomes and that IMPUTE v2 outperforms other methods in this setting at both rare and common SNPs, with overall error rates that are 15%–20% lower than those of the closest competing method. One particularly challenging aspect of next-generation association studies is to integrate information across multiple reference panels genotyped on different sets of SNPs; we show that our approach to this problem has practical advantages over other suggested solutions

Characterization of nanometer-sized, mechanically exfoliated graphene on the H-passivated Si(100) surface using scanning tunnelling microscopy

We have developed a method for depositing graphene monolayers and bilayers with minimum lateral dimensions of 2-10 nm by the mechanical exfoliation of graphite onto the Si(100)-2x1:H surface. Room temperature, ultra-high vacuum (UHV) tunnelling spectroscopy measurements of nanometer-sized single-layer graphene reveal a size dependent energy gap ranging from 0.1-1 eV. Furthermore, the number of graphene layers can be directly determined from scanning tunnelling microscopy (STM) topographic contours. This atomistic study provides an experimental basis for probing the electronic structure of nanometer-sized graphene which can assist the development of graphene-based nanoelectronics.Comment: Accepted for publication in Nanotechnolog

Iron-Fortified Drinking Water Studies for the Prevention of Children's Anemia in Developing Countries

Anemia and iron deficiency should receive special attention considering their high prevalence and serious consequences. For prevention, globally it is recommended to increase dietary iron intake, iron fortification of industrialized foods, and medical iron supplementation. Food fortification for the prevention of iron deficiency in developing countries should consider carriers locally available and consumed daily, requiring limited infrastructure and technology. Drinking water is the iron carrier we have been working for years for the prevention of iron deficiency and anemia in small children in Brazil. It was shown that studies with iron-fortified drinking water were proved to be effective on children's anemia prevention. Water is found everywhere, consumed daily by everyone may be easily fortified with simple technology, is low priced and was effective on the prevention of children's anemia. Fortification of drinking water with iron was locally implemented with the direct participation of the government and community. Government authorities, health personnel and population were part of the project and responsible for its community implementation. The mayor/municipality permitted and supported the proposal to supply it to children at their day-care centers. To keep the children drinking water iron fortified supply an officially authorized legislation was also approved

Copper deficiency anemia after bariatric surgery

Copper is a trace mineral essential to hematopoiesis and to the structure and function of the nervous system. Copper acts as a ligand to ferroxidase II, which oxidizes iron, allowing it to be mobilized and transported from the hepatic stores to the bone marrow to be used in erythropoiesis..Copper deficiency is a rare cause of anemia, leukopenia, and myeloneuropathy. In this report we describe a case of a 49 year-old overweight Caucasian woman who was referred June 2008 to the Obesity Outpatient Clinic at UNIFESP for evaluation of muscular pain and weakness in both legs, fatigue and chronic anemia. The past medical history included Scopinaro Surgery for severe obesity in November 2004 when her weight was approximately 180 Kg (BMI 68.6 kg/m2). She lost considerable weight after surgery, with her weight being stable around 80 kg (BMI 30). Because of clinical manifestations of sub nutrition, protein and vitamin deficiencies and severe anemia, which lead to several hospital admissions, in February 2008 she was submitted to another surgery from conversion of the gastrointestinal surgical Scopinaro procedure to that performed by Capella. She also had a history of hypothyroidism, hyperuricemia and panic syndrome, requiring specific medications, beyond parenteral iron and B12 vitamin therapies for anemia. Physical examination revealed a bilateral edema and no other abnormality. Laboratory tests revealed hypoalbuminemia and hypochromic megaloblastic anemia with normal serum levels of B12 vitamin, iron and ferritin. Due to the persistence of the neurologic symptoms and anemia in spite of adequate iron and B12 vitamin therapies, the hypothesis of copper deficiency was considered and a low serum level of 40μg/dl was found. An oral supplementation with high doses of copper resulted in evident clinical and laboratorial improvements after eight weeks of therapy. The diagnosis of copper deficiency has to be part of the differential diagnosis in patients with unexplained neurologic symptoms, anemia, and leukopenia, especially in those who underwent to previous gastrointestinal surgery