MBWA: A Massively Parallel Sequence Reads Aligner

This paper focuses on theoretical analysis, computational test and optimization of High Performance Computing Cluster (HPCC). Also known as Data Analytics Supercomputer (DAS), HPCC is built on Intel Many Integrated Core (MIC) for High Performance Linpack (HPL) test. Initially, a platform is configured by 5 nodes with an Intel® Xeon Phi <sup xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">™</sup> Coprocessor 31S1P per node to analyze power consumption as well as parallel level of Intel MIC accelerator. Compiling with Message Passing Interface (MPI) library and Math Kernel Library (MKL), the "Make" file is modified and debugged by adjusting parameters in hpccinf.txt. According to multiple Infiniband nodes evaluation, the libhpl library of Intel is employed and value of NB is set to be 960 for a single node with one MIC while debugging. Moreover, this paper optimized the algorithm on Double Precision General Matrix Multiplication (DGEMM) test and PTRANS to acquire efficient, precise and truncated working time duration. From HPL and Gridding Program experimental results, it's clear that the theoretical analysis and experiments were performed successfully.

Heterogeneous architectures, where a multicore processor, which is optimized for fast single-thread performance, is accompanied with a large number of simpler, but more power-efficient cores optimized for parallel workloads, such as NVIDIA's GPUs or Intel's Many Integrated Core (MIC), have been receiving a lot attention recently. Although NVIDIA's GPUs include built-in support for parallelism control, the MIC uses classical software thread creation and scheduling done by the operating system (OS). While efficient thread creation is desired in such many-core environments, current OS APIs provide the facility of creating only one thread at a time. In this paper, we propose a new system call for parallel thread creation on many-core coprocessors and show that it can perform up to 6.9 times better than the sequential version when executed on Intel's MIC software development platform.

The partitioned predictive vector quantization (PPVQ) algorithm is known for its high compression ratio for lossless compression of the ultraspectral sounder data with high spatial and spectral resolutions. With the advent of the multicore technologies, parallelization of several parts of the algorithm has been explored in previous work using a compute unified device architecture (CUDA) aided environment on the Graphics Processing Unit (GPU). Recently the Intel Many Integrated Core (MIC) architecture on a coprocessor is introduced which shows promise in handling more divergent workloads as needed in PPVQ. Therefore we will explore the parallel performance of the MIC-aided implementation. With parallelization of the two most time-consuming modules of linear prediction and vector quantization in PPVQ, the total processing time of an AIRS granule can be compressed in less than 7.5 seconds which is equivalent to a speedup of ~8.8x. The use of MIC for PPVQ compression is thus promising as a low-cost and effective compression solution for ultraspectral sounder data for ground rebroadcast use.

Sequence alignment is a fundamental task in bioinformatics, because many downstream applications rely on it. The recent emergence of the third-generation sequencing technology requires new sequence alignment algorithms that handle longer read lengths as well as more sequencing errors. Furthermore, the rapidly increasing volume of sequence data calls for efficient analysis solutions. To address this need, we propose to utilize commodity parallel processors to perform the long read alignment. Specifically, we propose manymap, an acceleration of the leading CPU-based long read aligner minimap2 on the CPU, the GPU, and the Intel Xeon Phi processor. We eliminate intra-loop data dependency in the base-level alignment step of the original minimap2 through redesigning memory layouts of dynamic programming (DP) matrices. This change facilitates the effective vectorization of the most time-consuming procedure in alignment. Additionally, we apply architecture-aware optimizations, such as utilizing high bandwidth memory on Xeon Phi and concurrent kernel execution on GPU. We evaluate our manymap in comparison with the extended minimap2 on a Xeon Gold 5115 CPU, a Tesla V100 GPU, and a Xeon Phi 7210 processor. Our results show that manymap outperforms minimap2 by up to 2.3 times on the overall execution time and 4.5 times on the base-level alignment step. © 2019 ACM.

Next generation sequencing (NGS) technologies have enabled cheap, large-scale, and high-throughput production of short DNA sequence reads and thereby have promoted the explosive growth of data volume. Unfortunately, the produced reads are short and prone to contain errors that are incurred during sequencing cycles. Both large data volume and sequencing errors have complicated the mapping of NGS reads onto the reference genome and have motivated the development of various aligners for very short reads, typically less than 100 base pairs (bps) in length. As read length continues to increase, propelled by advances in NGS technologies, these longer reads tend to have higher sequencing error rates and more true mutations (including substitutions, insertions, or deletions) to the genome. Such new characteristics make inefficient the aligners, which are optimized for very short reads and support only ungapped alignments or gapped alignments with very limited number of gaps (typically one gap), and thereby call for new aligners with fully gapped alignment supported. In this chapter, we present the CUSHAW software suite for NGS read alignment, which is open-source and consists of three individual aligners: CUSHAW, CUSHAW2, and CUSHAW3. This suite offers parallel and efficient NGS read alignments to large genomes, such as the human genome, by harnessing multi-core CPUs or compute unified device architecture (CUDA)-enabled graphics processing units (GPUs). Moreover, it has the capability to align both base-space and color-space reads and is consistently shown to be one of the best alignment tools through our performance evaluations.

A discord is a refinement of the concept of an anomalous subsequence of a time series. The task of discovering discords is applied in a wide range of subject areas involving time series: medicine, economics, climate modeling, and others. In this paper, we propose a novel parallel algorithm for discord discovery using Intel MIC (Many Integrated Core) accelerators in the case when time series fit in the main memory. We achieve parallelization through thread-level parallelism and OpenMP technology. The algorithm employs a set of matrix data structures to store and index the subsequences of a time series and to provide an efficient vectorization of computations on the Intel MIC platform. Moreover, the algorithm exploits the ability to independently computing Euclidean distances between subsequences of a time series. The algorithm iterates subsequences in two nested loops; it parallelizes the outer and the inner loops separately and differently, depending on both the number of running threads and the cardinality of the sets of subsequences scanned in the loop. The experimental evaluation shows the high scalability of the proposed algorithm.

The continuous development of sequencing technologies has enabled researchers to obtain large amounts of biological sequence data, and this has resulted in increasing demands for software that can perform sequence alignment fast and accurately. A number of algorithms and tools for sequence alignment have been designed to meet the various needs of biologists. Here, the ideas that prevail in the research of sequence alignment and some quality estimation methods for multiple sequence alignment tools are summarized.

In medical and genomic research, the Next Generation Sequencing (NGS) has a major role. Presently, NGS data are produced at the rate of 10 TB a day and challenge the storage and data processing capacities. These huge datasets are being used by a wide sort of applications such as customized cancer healing and precision medicine. NGS technologies offer prospects for understanding unidentified species and complex syndrome. To utilize genomic data for such applications, the genomic data in the form of short reads produced by NGS initially has to be assembled into whole genome sequence. And then, the sequences have to be compared for similarity and variation discovery which will be useful for analyzing and arriving at health-related solutions. In this paper, the fundamental methods for short read alignment such as assembly-based and alignment-based methods are discussed. Followed by which, the different ways to compare the sequences to check the alignment for similarity/dissimilarity discovery are discussed. This comparative analysis report can be utilized for health-related medical decisions.

The most widely used parallel architectures in today's High Performance Computing systems utilize multi-core CPUs, many-core GPUs or Intel's MIC (Many Integrated Core). The effort of new algorithm and implementation development greatly influences the performance on these architectures, and the differences between their underlying ILP parallelism - namely SIMT (Single Instruction Multiple Thread) and SIMD (Single Instruction Multiple Data) - require different approaches. The aim of the work to be presented is to show how high performance can be achieved in solving multiple scalar- and block-tridiagonal system of equations. The Thomas algorithm is implemented on all three hardware platforms, and for the GPU we also implement a hybrid algorithm based on Parallel Cyclic Reduction and Thomas algorithm for solving scalar problem and a thread level, work-sharing based algorithm for block-tridiagonal problems. Performance comparisons and a discussion on efficiency are also included.

Motivation and Objectives The output of a high throughput next generation sequencing (NGS) machine is a collection of short reads, which have to be properly assembled in order to reconstruct the original DNA sequence of the analyzed organism (Metzker, 2010; Earl, 2011). The DNA sequence assembly process is based on aligning and merging these reads for effectively reconstructing the real primary structure of the DNA sample sequence or reference genome. The use of NGS machines results in much larger sets of reads to be assembled, posing new problems for computer scientists and bioinformaticians. In particular, a relevant issue is related with the trade-off between precision of the assembly process and its computational time, stating the need for faster methods that can keep pace with the speed and volume of reads that are generated with NGS. An important step in DNA assembly is the identification of a subset of read pairs that have a high probability of being aligned sequentially in the reconstruction. Such step is often referred to as filtering, and amounts in selecting a significantly smaller subset of the initial set of read pairs (whose dimension is quadratic in the number of initial reads) that can be then processed by an alignment algorithm, usually quite time consuming. The desired effect of filtering is then to quickly filter out from the candidate set of read pairs those that would not provide a good alignment in the following phase. The computation cost of filtering should then be balanced by the speed-up obtained when a smaller set of read pairs is considered for alignment. In this work we propose and test the use of alignment free distances to evaluate the similarity between two short reads as a technique for filtering good read pairs to be assembled. The method operates in constant time in the string length and is tested in its ability to emulate, with a proper level of precision, much more time consuming methods to evaluate the similarity between short DNA sequences, such as the established Needleman-Wunsch edit distance (Needleman, Wunsch, and Christian, 1970), often used in the final step of the assembly procedure. These preliminary experiments show the efficacy of this approach for filtering the promising read pairs - eligible candidates to successfully assemble the entire genome of a given organism. Therefore, the alignment free reads filtering may significantly accelerate the assembly process without a substantial loss in accuracy of the DNA sample sequence reconstruction. Methods ODNA sequence assembly The DNA sequence assembly process is based on the alignment and merging of reads (stretch of sequences) in order to reconstruct the original primary structure of the DNA sample sequences. Given a set of sequences S = {s1, s2, …, sn}, where s ∈ S is a fragment of the primary structure of DNA (read) (e.g. s = {ATTCGA...CTGACT}), assembly is in charge of building the longest sequence from the set S where each pair of consequent reads obey certain similarity conditions. DNA read pairs filtering and Alignment Free Distance This step identifies the promising read pairs in order to reduce the amount of input data given to the real assembly algorithm. We adopted a very quick measure of the similarity between two reads, Alignment Free (AF) based distance (Vinga and Almeida, 2003). AF computes the similarity of two strings based only on the dictionary of their substrings, irrespective of their relative position. As a dictionary we considered the set of 4-mer (sequences composed of 4 different nucleotides) and then built a profile for each read composed by the relative frequencies of each 4-mer in the read. The Euclidean distance between the profiles of two reads was taken as an inverse measure of the similarity of the two reads and thus as an indication that the two reads formed a promising pair to be considered in the assembly phase. AF filtering was then used defining a proper threshold on the AF distance and discarding all the pairs that exhibited a AF distance above the threshold. Computational complexity of AF distance is a constant linearly bounded by the number of k-mers adopted and the length of the strings to be compared. Comparing with other distances: Needleman-Wunsch and “Bowtie” distance Along with AF we considered the well-established Needleman-Wunsch edit distance (NW) and compared them in their ability to identify significant pairs. This comparison was based on the computation of a sort of perfect distance computed after an alignment over an already known sequence has been performed. Such distance, referred to as Bowtie distance (BT), was obtained as follows: a. a large number of reads coming from a known sequence were considered; b. hese reads were aligned over the known sequence using the standard Bowtie algorithm (Langmead et al, 2009); c. any two reads received a maximum BT distance if their alignment did not intersect over the reference sequence, else they received a distance inversely proportional to their intersection over the sequence (e.g., they would have BT distance equal to 0 if they were aligned one on top (or inside) of the other by the Bowtie algorithm). By construction we assumed BT distance to be the reference distance, e.g., the distance that expressed the best possible alignments - being based on the knowledge of the reference sequence - and tested the correlation of AF and NW with BT; moreover, we verified the ability of AF and NW to predict that a given read pair had BT distance above or below a given threshold. Results and Discussion For our test we considered the E.Coli genome and a set of reads from this genome reads obtained by Roche 454 sequencing machine. Reads have average length of ~235 nucleotides and standard deviation of approx. 10 (the large majority of them having length in the interval 225-245). Reads were aligned with the reference sequence with Bowtie and then 100,000 were sampled at random according to their alignment along the sequence. Reads were considered both forward and reversed, giving rise to a total of 200,0002 read pairs. All 620,798 read pairs with BT distance &lt; 1 were considered for the experiments; then, out of the remaining pairs, 233,099 were sampled at random. A total of 853,897 read pairs composed the working data set. For all these reads, NW distance and AF distance over the 4-mer were computed. AF, NW and BT distances were all normalized between 0 (maximal similarity) and 1 (maximal dissimilarity). The first interesting results was that the correlation between distances showed that AF approximates BT somehow better than NW: we obtained a correlation coefficient of 0.761 for AF and BT, compared with a smaller 0.706 when NW and BT were considered (coherently, correlation between AF and NW is 0,721). The second interesting results was obtained when we compared the ability of AF and NW to predict whether BT was above or below a given threshold. We defined a threshold predictor for a given function F2 based on function F1 and on a given pair a1, a2 as follows: if (F1 &lt; a1) then predict (F2 &lt; a2), else predict (F2 ≥ a2). To a given pair (a1, a2), we associated the measure of True Positive rate (TP) (percentage of cases where (F1 &lt; a1) and (F2 &lt; a2) and of True Negative rate (TN) (percentage of cases where (F1 ≥ a1) and (F2 ≥ a2); analogously we defined False Positive rate (FP) and False Negative rate (FN). For each (a1, a2) with both values ranging from 0 to 1, we then computed, with step 0.05, the positive and negative error rates taking AF as a predictor of BT and NW as a predictor of BT. Part of the results are summarized in the charts of Figure 1, that show for 3 different levels of a2 (0.1, 0.2, and 0.3) the precision of the predictors (y-axis) when the value of a1 is changed (x-axis), both when AF is used as a predictor of BT (charts in the first row) and when NW is used as a predictor of BT (charts in second row). Similar results are obtained also for other levels of a2, here omitted for brevity. The curves bring to light very clearly how AF is a very good threshold predictor for BT for the considered data; despite its light computational complexity, it appears to perform significantly better than the more complex NW edit distance when its ability to support a threshold predictor is considered. Acknowledgements The authors are partially supported by the FLAGSHIP ”InterOmics” project (PB.P05) funded by the Italian MIUR and CNR institutions, and by the cooperative programme 2010–2012 between the National Research Council of Italy (CNR) and the Polish Academy of Sciences (PAN). References Earl D et al (2011); Assemblathon 1: A competitive assessment of de novo short read assembly methods; Genome Research, 21 Langmead B, Trapnell C, et al (2009); Ultrafast and memory-efficient alignment of short DNA sequences to the human genome; Genome Biology, 10:R25 Metzker ML (2010); Sequencing technologies — the next generation; Nat Rev Genet., 11(1) Needleman SB, Wunsch CD. (1970). A general method applicable to the search for similarities in the amino acid sequence of two proteins. Journal of Molecular Biology 48(3): 443–53 Vinga S, Almeida J (2003); Alignment-free sequence comparison—a review, Bioinformatics 19 (4), 513-523 Note: Figures and tables are available in PDF version only.

Sensitive detection of internal tandem duplication (ITD) mutation of FLT3 is very important in acute myeloid leukemia. To increase detection sensitivity of FLT3-ITD, we developed new detection algorithm using next generation sequencing (NGS) data. We validated results using nested polymerase chain reaction (PCR) methods. We compared results of NGS data, nested PCR and conventional PCR methods. First, using whole exome sequencing data of 83 AML patients, we applied calling algorithm for FLT3-ITD. Briefly, to detect ITDs with NGS data, the reads are aligned to a reference sequence (UCSC hg19), with BWA which is a read aligner allowing soft-clipping. Some reads can be an indication of the occurrence of ITD and BWA aligns those reads as soft-clipped. Second, we deigned two types of primer for Nested PCR. The first primer was targeted wildly for between exon14 and exon15 of FLT3 gene. Nested PCR primer was deigned to target previously reported regions which are frequently occurred ITD mutation. PCR reactions of two steps were performed using the PCR primers sequentially. In these 83 patients, FLT3-ITD was positive only in 7 patients when tested by conventional PCR methods. When NGS detection method was applied, this resulted in positive FLT3-ITD in 11 patients (11/83, 13%). When validation was performed using nested PCR, FLT3-ITD was confirmed in all of 11 patients. Nested PCR detected additional 4 patient positive for FLT3-ITD in this population. For 68 patients, FLT3-ITD was negative by both NGS and nested PCR method. Overall, NGS method improved sensitivity of FLT3-ITD detection by 57% in this population. And the concordance rate of NGS method and nested PCR was 95.2% (79/83). Then we investigated clinical significance of sensitive FLT3-ITD detection. For this, we performed nested PCR and conventional PCR at the same time in 238 AML patients to detect FLT3-ITD. Positive rate for FLT3-ITD was 20% (48/238) and 10% (24/238) by nested PCR and conventional PCR respectively. When survival analysis was performed, among patients with negative FLT3-ITD result by conventional PCR, patients who showed positive for FLT3-ITD by nested PCR had shorter overall survival compared to those who showed negative for FLT3-ITD by nested PCR. (p = 0.03). This implies that sensitive FLT3-ITD detection using nested PCR is clinically meaningful. Diagnosis of FLT3-ITD is very important genetic factor, leading a therapeutic direction for AML patient. Here we report that we have developed alternative more sensitive detection methods for FLT3-ITD based on nested PCR and NGS. Sensitive detection of FLT3-ITD was clinically meaningful, suggesting that these methods should be incorporated in a future clinical practice. Also, we want to note that, NGS method is capable of quantifying FLT3-ITD size and amount in AML patients. Citation Format: Daeyoon Kim, Yoojin Hong, Youngil Koh, Sung-Soo Yoon, Choong-Hyun Sun, Kwang-Sung Ahn, Seungmook Lee, Hongseok Yun, Suyeon Lee. Improved sensitive detection method of FLT3 (FMS-like tyrosine kinase) internal tandem duplication (ITD) mutation using next-generation sequencing technology and nested PCR. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3186.

During 2012, next generation sequencing (NGS) has attracted great attention in the biomedical research community, especially for personalized medicine. Also, third generation sequencing has become available. Therefore, state-of-art sequencing technology and analysis are reviewed in this Bioinformatics spotlight on 2012. Next-generation sequencing (NGS) is high-throughput nucleic acid sequencing technology with wide dynamic range and single base resolution. The full promise of NGS depends on the optimization of NGS platforms, sequence alignment and assembly algorithms, data analytics, novel algorithms for integrating NGS data with existing genomic, proteomic, or metabolomic data, and quantitative assessment of NGS technology in comparing to more established technologies such as microarrays. NGS technology has been predicated to become a cornerstone of personalized medicine. It is argued that NGS is a promising field for motivated young researchers who are looking for opportunities in bioinformatics.

Next Generation Sequencing (NGS) platforms typically produce short reads of size 50-150 base pairs (bp). The number of such short reads can be up to 6 billion per run. To align these short reads to a large genome is a computationally challenging problem. In this paper, we address this problem by considering the design and optimization of parallel sequence alignment on GPU based hybrid architectures. Even though the sequence alignment algorithm is inherently data-parallel, issues such as (a) space-time trade-offs in the Indexing schema, (b) need for fast candidate location search (CAL) on GPU, (c) maintaining low divergence along with low space for the dynamic programming based local alignment, make this a very challenging problem. We present the design of our novel parallel algorithm Graphics processor Accelerated BFAST (GrABFAST) for large scale read alignment that overcomes these challenges and demonstrates superior performance compared to Intel multi-core architectures. Using 5 large genomes including those of Humans, Maize, Horse, Dog and Bacteria, we demonstrate a speedup of around 6x using Fermi Tesla C2070 GPUs vs the BFAST algorithm on 16 core Intel Xeon 5570 architecture.

In the next generation sequencing (NGS) read alignment problem, millions of deoxyribonucleic acid (DNA) fragments, called reads, are mapped to a reference genome. Read alignment is typically carried out using traditional computing platforms, which have become a limiting factor in the speed of the process. The massive scale of the problem makes it an attractive target for acceleration. In this paper, we design a read alignment algorithm designed to run on a heterogeneous system composed of a graphics processing unity (GPU) and a multicore central processing unit (CPU). We introduce novel techniques for the alignment process and construct a computational pipeline of overlapped CPU and GPU stages. We compare our tool with the BWA-mem alignment tool, and the results show substantial speedups.

CRISPR/Cas9-based genome editing has yet to be reported in species of the Platyhelminthes. We tested this approach by targeting omega-1 (ω1) of Schistosoma mansoni as proof of principle. This secreted ribonuclease is crucial for Th2 polarization and granuloma formation. Schistosome eggs were exposed to Cas9 complexed with guide RNA complementary to ω1 by electroporation or by transduction with lentiviral particles. Some eggs were also transfected with a single stranded donor template. Sequences of amplicons from gene-edited parasites exhibited Cas9-catalyzed mutations including homology directed repaired alleles, and other analyses revealed depletion of ω1 transcripts and the ribonuclease. Gene-edited eggs failed to polarize Th2 cytokine responses in macrophage/T-cell co-cultures, while the volume of pulmonary granulomas surrounding ω1-mutated eggs following tail-vein injection into mice was vastly reduced. Knock-out of ω1 and the diminished levels of these cytokines following exposure showcase the novel application of programmed gene editing for functional genomics in schistosomes.