Displaying publications 81 - 100 of 216 in total

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  1. Fulltext Advances in the study of nodavirus
    Yong CY, Yeap SK, Omar AR, Tan WS
    PeerJ, 2017;5:e3841.
    PMID: 28970971 DOI: 10.7717/peerj.3841
    Nodaviruses are small bipartite RNA viruses which belong to the family of Nodaviridae. They are categorized into alpha-nodavirus, which infects insects, and beta-nodavirus, which infects fishes. Another distinct group of nodavirus infects shrimps and prawns, which has been proposed to be categorized as gamma-nodavirus. Our current review focuses mainly on recent studies performed on nodaviruses. Nodavirus can be transmitted vertically and horizontally. Recent outbreaks have been reported in China, Indonesia, Singapore and India, affecting the aquaculture industry. It also decreased mullet stock in the Caspian Sea. Histopathology and transmission electron microscopy (TEM) are used to examine the presence of nodaviruses in infected fishes and prawns. For classification, virus isolation followed by nucleotide sequencing are required. In contrast to partial sequence identification, profiling the whole transcriptome using next generation sequencing (NGS) offers a more comprehensive comparison and characterization of the virus. For rapid diagnosis of nodavirus, assays targeting the viral RNA based on reverse-transcription PCR (RT-PCR) such as microfluidic chips, reverse-transcription loop-mediated isothermal amplification (RT-LAMP) and RT-LAMP coupled with lateral flow dipstick (RT-LAMP-LFD) have been developed. Besides viral RNA detections, diagnosis based on immunological assays such as enzyme-linked immunosorbent assay (ELISA), immunodot and Western blotting have also been reported. In addition, immune responses of fish and prawn are also discussed. Overall, in fish, innate immunity, cellular type I interferon immunity and humoral immunity cooperatively prevent nodavirus infections, whereas prawns and shrimps adopt different immune mechanisms against nodavirus infections, through upregulation of superoxide anion, prophenoloxidase, superoxide dismutase (SOD), crustin, peroxinectin, anti-lipopolysaccharides and heat shock proteins (HSP). Potential vaccines for fishes and prawns based on inactivated viruses, recombinant proteins or DNA, either delivered through injection, oral feeding or immersion, are also discussed in detail. Lastly, a comprehensive review on nodavirus virus-like particles (VLPs) is presented. In recent years, studies on prawn nodavirus are mainly focused on Macrobrachium rosenbergii nodavirus (MrNV). Recombinant MrNV VLPs have been produced in prokaryotic and eukaryotic expression systems. Their roles as a nucleic acid delivery vehicle, a platform for vaccine development, a molecular tool for mechanism study and in solving the structures of MrNV are intensively discussed.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  2. Disruption of methicillin-resistant Staphylococcus aureus protein synthesis by tannins
    Adnan SN, Ibrahim N, Yaacob WA
    Germs, 2017 Dec;7(4):186-192.
    PMID: 29264356 DOI: 10.18683/germs.2017.1125
    Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) is a worldwide public health threat, displaying multiple antibiotic resistance that causes morbidity and mortality. Management of multidrug-resistant (MDR) MRSA infections is extremely difficult due to their inherent resistance to currently used antibiotics. New antibiotics are needed to combat the emergence of antimicrobial resistance.

    Methods: The in vitro effect of tannins was studied against MRSA reference strain (ATCC 43300) and MRSA clinical strains utilizing antimicrobial assays in conjunction with both scanning and transmission electron microscopy. To reveal the influence of tannins in MRSA protein synthesis disruption, we utilized next-generation sequencing (NGS) to provide further insight into the novel protein synthesis transcriptional response of MRSA exposed to these compounds.

    Results: Tannins possessed both bacteriostatic and bactericidal activity with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 0.78 and 1.56 mg/mL, respectively, against all tested MRSA. Scanning and transmission electron microscopy of MRSA treated with tannins showed decrease in cellular volume, indicating disruption of protein synthesis.

    Conclusion: Analysis of a genome-wide transcriptional profile of the reference strain ATCC 43300 MRSA in response to tannins has led to the finding that tannins induced significant modulation in essential ribosome pathways, which caused a reduction in the translation processes that lead to inhibition of protein synthesis and obviation of bacterial growth. These findings highlight the potential of tannins as new promising anti-MRSA agents in clinical application such as body wash and topical cream or ointments.

    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  3. Fulltext Finding Nemo: hybrid assembly with Oxford Nanopore and Illumina reads greatly improves the clownfish (Amphiprion ocellaris) genome assembly
    Tan MH, Austin CM, Hammer MP, Lee YP, Croft LJ, Gan HM
    Gigascience, 2018 03 01;7(3):1-6.
    PMID: 29342277 DOI: 10.1093/gigascience/gix137
    Background: Some of the most widely recognized coral reef fishes are clownfish or anemonefish, members of the family Pomacentridae (subfamily: Amphiprioninae). They are popular aquarium species due to their bright colours, adaptability to captivity, and fascinating behavior. Their breeding biology (sequential hermaphrodites) and symbiotic mutualism with sea anemones have attracted much scientific interest. Moreover, there are some curious geographic-based phenotypes that warrant investigation. Leveraging on the advancement in Nanopore long read technology, we report the first hybrid assembly of the clown anemonefish (Amphiprion ocellaris) genome utilizing Illumina and Nanopore reads, further demonstrating the substantial impact of modest long read sequencing data sets on improving genome assembly statistics.

    Results: We generated 43 Gb of short Illumina reads and 9 Gb of long Nanopore reads, representing approximate genome coverage of 54× and 11×, respectively, based on the range of estimated k-mer-predicted genome sizes of between 791 and 967 Mbp. The final assembled genome is contained in 6404 scaffolds with an accumulated length of 880 Mb (96.3% BUSCO-calculated genome completeness). Compared with the Illumina-only assembly, the hybrid approach generated 94% fewer scaffolds with an 18-fold increase in N50 length (401 kb) and increased the genome completeness by an additional 16%. A total of 27 240 high-quality protein-coding genes were predicted from the clown anemonefish, 26 211 (96%) of which were annotated functionally with information from either sequence homology or protein signature searches.

    Conclusions: We present the first genome of any anemonefish and demonstrate the value of low coverage (∼11×) long Nanopore read sequencing in improving both genome assembly contiguity and completeness. The near-complete assembly of the A. ocellaris genome will be an invaluable molecular resource for supporting a range of genetic, genomic, and phylogenetic studies specifically for clownfish and more generally for other related fish species of the family Pomacentridae.

    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  4. Fulltext Deep sequencing and in silico analysis of small RNA library reveals novel miRNA from leaf Persicaria minor transcriptome
    Samad AFA, Nazaruddin N, Murad AMA, Jani J, Zainal Z, Ismail I
    3 Biotech, 2018 Mar;8(3):136.
    PMID: 29479512 DOI: 10.1007/s13205-018-1164-8
    In current era, majority of microRNA (miRNA) are being discovered through computational approaches which are more confined towards model plants. Here, for the first time, we have described the identification and characterization of novel miRNA in a non-model plant, Persicaria minor (P. minor) using computational approach. Unannotated sequences from deep sequencing were analyzed based on previous well-established parameters. Around 24 putative novel miRNAs were identified from 6,417,780 reads of the unannotated sequence which represented 11 unique putative miRNA sequences. PsRobot target prediction tool was deployed to identify the target transcripts of putative novel miRNAs. Most of the predicted target transcripts (mRNAs) were known to be involved in plant development and stress responses. Gene ontology showed that majority of the putative novel miRNA targets involved in cellular component (69.07%), followed by molecular function (30.08%) and biological process (0.85%). Out of 11 unique putative miRNAs, 7 miRNAs were validated through semi-quantitative PCR. These novel miRNAs discoveries in P. minor may develop and update the current public miRNA database.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  5. Fulltext The clinical impact of using complex molecular profiling strategies in routine oncology practice
    Laes JF, Aftimos P, Barthelemy P, Bellmunt J, Berchem G, Camps C, et al.
    Oncotarget, 2018 Apr 17;9(29):20282-20293.
    PMID: 29755651 DOI: 10.18632/oncotarget.24757
    Molecular profiling and functional assessment of signalling pathways of advanced solid tumours are becoming increasingly available. However, their clinical utility in guiding patients' treatment remains unknown. Here, we assessed whether molecular profiling helps physicians in therapeutic decision making by analysing the molecular profiles of 1057 advanced cancer patient samples after failing at least one standard of care treatment using a combination of next-generation sequencing (NGS), immunohistochemistry (IHC) and other specific tests. The resulting information was interpreted and personalized treatments for each patient were suggested. Our data showed that NGS alone provided the oncologist with useful information in 10-50% of cases (depending on cancer type), whereas the addition of IHC/other tests increased extensively the usefulness of the information provided. Using internet surveys, we investigated how therapy recommendations influenced treatment choice of the oncologist. For patients who were still alive after the provision of the molecular information (76.8%), 60.4% of their oncologists followed report recommendations. Most treatment decisions (93.4%) were made based on the combination of NGS and IHC/other tests, and an approved drug- rather than clinical trial enrolment- was the main treatment choice. Most common reasons given by physicians to explain the non-adherence to recommendations were drug availability and cost, which remain barriers to personalised precision medicine. Finally, we observed that 27% of patients treated with the suggested therapies had an overall survival > 12 months. Our study demonstrates that the combination of NGS and IHC/other tests provides the most useful information in aiding treatment decisions by oncologists in routine clinical practice.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  6. Improved nucleic acid extraction protocols for Ganoderma boninense, G. miniatocinctum and G. tornatum
    Nagappan J, Chin CF, Angel LPL, Cooper RM, May ST, Low EL
    Biotechnol Lett, 2018 Dec;40(11-12):1541-1550.
    PMID: 30203158 DOI: 10.1007/s10529-018-2603-7
    The first and most crucial step of all molecular techniques is to isolate high quality and intact nucleic acids. However, DNA and RNA isolation from fungal samples are usually difficult due to the cell walls that are relatively unsusceptible to lysis and often resistant to traditional extraction procedures. Although there are many extraction protocols for Ganoderma species, different extraction protocols have been applied to different species to obtain high yields of good quality nucleic acids, especially for genome and transcriptome sequencing. Ganoderma species, mainly G. boninense causes the basal stem rot disease, a devastating disease that plagues the oil palm industry. Here, we describe modified DNA extraction protocols for G. boninense, G. miniatocinctum and G. tornatum, and an RNA extraction protocol for G. boninense. The modified salting out DNA extraction protocol is suitable for G. boninense and G. miniatocinctum while the modified high salt and low pH protocol is suitable for G. tornatum. The modified DNA and RNA extraction protocols were able to produce high quality genomic DNA and total RNA of ~ 140 to 160 µg/g and ~ 80 µg/g of mycelia respectively, for Single Molecule Real Time (PacBio Sequel® System) and Illumina sequencing. These protocols will benefit those studying the oil palm pathogens at nucleotide level.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  7. Bacterial regulatory RNAs: complexity, function, and putative drug targeting
    Cheah HL, Raabe CA, Lee LP, Rozhdestvensky TS, Citartan M, Ahmed SA, et al.
    Crit Rev Biochem Mol Biol, 2018 08;53(4):335-355.
    PMID: 29793351 DOI: 10.1080/10409238.2018.1473330
    Over the past decade, RNA-deep sequencing has uncovered copious non-protein coding RNAs (npcRNAs) in bacteria. Many of them are key players in the regulation of gene expression, taking part in various regulatory circuits, such as metabolic responses to different environmental stresses, virulence, antibiotic resistance, and host-pathogen interactions. This has contributed to the high adaptability of bacteria to changing or even hostile environments. Their mechanisms include the regulation of transcriptional termination, modulation of translation, and alteration of messenger RNA (mRNA) stability, as well as protein sequestration. Here, the mechanisms of gene expression by regulatory bacterial npcRNAs are comprehensively reviewed and supplemented with well-characterized examples. This class of molecules and their mechanisms of action might be useful targets for the development of novel antibiotics.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  8. Fulltext Transcriptome data of the carrageenophyte Eucheuma denticulatum
    Othman R, Abd Rasib AA, Ilias MA, Murthy S, Ismail N, Mohd Hanafi N
    Data Brief, 2019 Jun;24:103824.
    PMID: 30984808 DOI: 10.1016/j.dib.2019.103824
    Eucheuma denticulatum or commonly known as "Spinosum", is an economically important red alga that naturally grows on coral reefs with moderately strong currents in tropical and sub-tropical areas. This species is the primary source of iota-carrageenan which has high demands in the food, pharmaceutical and manufacturing industries, and as such it has been widely cultivated. The increasing global demand for carrageenan has led to extensive commercial cultivation of carrageenophytes mainly in the tropics. The carrageenophyte seaweeds including E. denticulatum are indigenous to Sabah, Malaysia. To enrich the information on the genes involved in carrageenan biosynthesis, RNA sequencing has been performed and transcriptomic dataset has been generated using Illumina HiSeq™ 2000 sequencer. The raw data and transcriptomic data have been deposited in NCBI database with the accession number PRJNA477734. These data will provide valuable resources for functional genomics annotation and investigation of mechanisms underlying the regulations of genes in this algal species.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  9. Assessing diet of the rufous-winged philentoma (philentoma pyrhoptera) in lowland tropical forest using next-generation sequencing
    Mohammad Saiful Mansor, Shukor Md. Nor, Rosli Ramli
    Sains Malaysiana, 2018;47:1045-1050.
    Dietary study provides understanding in predator-prey relationships, yet diet of tropical forest birds is poorly understood.
    In this study, a non-invasive method, next-generation sequencing (Illumina MiSeq platform) was used to identify prey in
    the faecal samples of the Rufous-winged Philentoma (Philentoma pyrhoptera). Dietary samples were collected in lowland
    tropical forest of central Peninsular Malaysia. A general invertebrate primer pair was used for the first time to assess
    diet of tropical birds. The USEARCH was used to cluster the COI mtDNA sequences into Operational Taxonomic Unit (OTU).
    OTU sequences were aligned and queried through the GenBank or Biodiversity of Life Database (BOLD). We identified
    26 distinct arthropod taxa from 31 OTUs. Of all OTUs, there was three that could be identified up to species level, 20 to
    genus level, three to family level and five could not assigned to any taxa (the BLAST hits were poor). All sequences were
    identified to class Insecta belonging to 18 families from four orders, where Lepidoptera representing major insect order
    consumed by study bird species. This non-invasive molecular approach provides a practical and rapid technique to
    understand of how energy flows across ecosystems. This technique could be very useful to screen for possible particular
    pest insects consumed by insectivores (e.g. birds and bats) in crop plantation. A comprehensive arthropod studies and
    local reference sequences need to be added to the database to improve the proportion of sequences that can be identified.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  10. Mitogenome of gymnothorax minor and phylogenetic relationship with its congeners and related genera (anguilliformes: muraenidae)
    Sze-Looi Song, Kar-Hoe Loh, Phaik-Eem Lim, Amy Yee-Hui Then, Hoi-Sen Yong, Praphathip Eamsobhana
    Sains Malaysiana, 2018;47:2519-2531.
    Gymnothorax minor is a moray eel of the family Muraenidae found in the Western Pacific Ocean. We report here
    its complete mitogenome as determined by Illumina next-generation sequencing and the phylogenetic relationship
    with its congeners and other taxa of the family Muraenidae. The whole mitogenome of G. minor had a total length
    of 16,574 bp, comprising 37 genes - 13 protein-coding genes (PCGs), two ribosomal ribonucleic acid (rRNA) and 22
    transfer ribonucleic acid (tRNA) genes - and a control region. Excepting cox1 with GTG, the other 12 PCGs had ATG
    start codon. Seven of its PCGs had incomplete stop codon - five (nad2; cox1; cox2; nad3 and nad4) with T and two
    (atp6 and cox3) with TA. Molecular phylogeny based on 13 PCGs was concordant with 15 mitochondrial genes (13 PCGs
    and 2 rRNA genes). The subfamily Muraeninae as well as the subfamily Uropterygiinae were monophyletic. However,
    the genus Gymnothorax was paraphyletic, with G. minor forming a sister group with Rhinomuraena quaesita in the
    lineage containing also G. kidako and G. formosus forming a sister group with Enchelynassa canina. The phylogenetic
    relationship of the genus Gymnothorax and related taxa of the family Muraenidae, based on the mitochondrial cob
    gene, was in general similar to that based on 15 mt-genes. The mitogenome is useful for future studies on phylogenetics
    and systematics of eels of the family Muraenidae and other taxa of the order Anguilliformes.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  11. A phylogenomic analysis of Nepenthes (Nepenthaceae)
    Murphy B, Forest F, Barraclough T, Rosindell J, Bellot S, Cowan R, et al.
    Mol Phylogenet Evol, 2020 03;144:106668.
    PMID: 31682924 DOI: 10.1016/j.ympev.2019.106668
    Nepenthaceae is one of the largest carnivorous plant families and features ecological and morphological adaptations indicating an impressive adaptive radiation. However, investigation of evolutionary and taxonomic questions is hindered by poor phylogenetic understanding, with previous molecular studies based on limited loci and taxa. We use high-throughput sequencing with a target-capture methodology based on a 353-loci, probe set to recover sequences for 197 samples, representing 151 described or putative Nepenthes species. Phylogenetic analyses were performed using supermatrix and maximum quartet species tree approaches. Our analyses confirm five Western outlier taxa, followed by N. danseri, as successively sister to the remainder of the group. We also find mostly consistent recovery of two major Southeast Asian clades. The first contains common or widespread lowland species plus a Wallacean-New Guinean clade. Within the second clade, sects. Insignes and Tentaculatae are well supported, while geographically defined clades representing Sumatra, Indochina, Peninsular Malaysia, Palawan, Mindanao and Borneo are also consistently recovered. However, we find considerable conflicting signal at the site and locus level, and often unstable backbone relationships. A handful of Bornean taxa are inconsistently placed and require further investigation. We make further suggestions for a modified infra-generic classification of genus Nepenthes.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  12. Fulltext The de novo transcriptome of workers head of the higher group termite Globitermes sulphureus Haviland (Blattodea: Termitidae)
    Hussin NA, Najimudin N, Ab Majid AH
    Heliyon, 2019 Dec;5(12):e02969.
    PMID: 31872129 DOI: 10.1016/j.heliyon.2019.e02969
    The subterranean termite Globitermus sulphureus is an important Southeast Asian pest with limited genomic resources that causes damages to agriculture crops and building structures. Therefore, the main goal of this study was to survey the G. sulphureus transcriptome composition. Here, we performed de novo transcriptome for G. sulphureus workers' heads using Illumina HiSeq paired-end sequencing technology. A total of 88, 639, 408 clean reads were collected and assembled into 243, 057 transcripts and 193, 344 putative genes. The transcripts were annotated with the Trinotate pipeline. In total, 27, 061 transcripts were successfully annotated using BLASTX against the SwissProt database and 17, 816 genes were assigned to 47, 598 GO terms. We classified 14, 223 transcripts into COG classification, resulting in 25 groups of functional annotations. Next, a total of 12, 194 genes were matched in the KEGG pathway and 392 metabolic pathways were predicted based on the annotation. Moreover, we detected two endogenous cellulases in the sequences. The RT-qPCR analysis showed that there were significant differences in the expression levels of two genes β-glucosidase and endo-β-1,4-glucanase between worker and soldier heads of G. sulphureus. This is the first study to characterize the complete head transcriptome of a higher termite G. sulphureus using a high-throughput sequencing. Our study may provide an overview and comprehensive molecular resource for comparative studies of the transcriptomics and genomics of termites.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  13. Fulltext Complete genome sequence of fowl adenovirus-8b UPM04217 isolate associated with the inclusion body hepatitis disease in commercial broiler chickens in Malaysia reveals intermediate evolution
    Mat Isa N, Mohd Ayob J, Ravi S, Mustapha NA, Ashari KS, Bejo MH, et al.
    Virusdisease, 2019 Sep;30(3):426-432.
    PMID: 31803810 DOI: 10.1007/s13337-019-00530-9
    The main aim of our study was to explore the genome sequence of the inclusion body hepatitis associated Fowl adenovirus serotype 8b (FAdV-8b) UPM04217 and to study its genomic organisation. The nucleotide sequence of the whole genome of FAdV-8b UPM04217 was determined by using the 454 Pyrosequencing platform and the Sanger sequencing method. The complete genome was found to be 44,059 bp long with 57.9% G + C content and shared 97.5% genome identity with the reference FAdV-E genome (HG isolate). Interestingly, the genome analysis using ORF Finder, Glimmer3 and FGENESV predicted a total of 39 open reading frames (ORFs) compared to the FAdV-E HG that possessed 46 ORFs. Fourteen ORFs located within the central genomic region and 16 ORFs located within the left and right ends of the genome were assigned as being the high protein-coding regions. The fusion of the small ORFs at the right end terminal specifically in ORF22 and ORF33 could be the result of gene truncation in the FAdV-E HG. The frame shift mutation in ORF25 and other mutations in ORF13 and ORF17 might have lead to the emergence of genes that could have different functions. Besides, one of the minor capsid components, pVI, in FAdV-8b UPM04217 shared the highest similarity of 93% with that of FAdV-D, while only 47% similarity was found with FAdV-E. From the gene arrangement layout of the FAdV genome, FAdV-8b UPM04217 showed intermediate evolution between the FAdV-E HG and the FAdV-D although it was apparently more similar to the FAdV-E HG.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  14. Fulltext RNA-seq data from whole rice grains of pigmented and non-pigmented Malaysian rice varieties
    Zainal-Abidin RA, Zainal Z, Mohamed-Hussein ZA, Abu-Bakar N, Ab Razak MSF, Simoh S, et al.
    Data Brief, 2020 Jun;30:105432.
    PMID: 32280737 DOI: 10.1016/j.dib.2020.105432
    Pigmented rice is enriched with antioxidants, macro- and micronutrients. A comprehensive investigation of the gene expression patterns among the pigmented rice varieties would help to understand the cellular mechanism and biological processes of rice grain pigmentation. Hence, we performed RNA sequencing and analysis on the whole grain of dehusked mature seeds of selected six Malaysian rice varieties with varying grain pigmentations. These varieties were black rice (BALI and Pulut Hitam 9), red rice (MRM16 and MRQ100) and white rice (MR297 and MRQ76). Illumina HiSeq™ 4000 sequencer was used to generate total raw nucleotides of approximately 53 Gb in size. From 353,937,212 total paired-end raw reads, 340,131,496 total clean reads were obtained. The raw reads were deposited into European Nucleotide Archive (ENA) database and can be accessed via accession number PRJEB34340. This dataset allows us to identify and profile all expressed genes with functions related to nutritional traits (i.e. antioxidants, folate and amylose content) and quality trait (i.e. aroma) across both pigmented and non-pigmented rice varieties. In addition, the transcriptome data obtained will be valuable for discovery of potential gene markers and functional SNPs related to functional traits to assist in rice breeding programme.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  15. Producing desulfurized biogas using two-stage domesticated shear-loop anaerobic contact stabilization system
    Tijani H, Yuzir A, Abdullah N
    Waste Manag, 2018 Aug;78:770-780.
    PMID: 32559969 DOI: 10.1016/j.wasman.2018.06.045
    In this study, a two-stage domesticated shear-loop anaerobic contact stabilization (SLACS) system is introduced as a new reactor design to enhance methane productivity with significant reduction in hydrogen sulphide (H2S) synthesis. Due to the rich sulfate content in industrial wastewaters, the initial fermentation phase of anaerobic digestion is highly acidifying and often leads to severe performance losses, digester's instability, and even culture crash. The SLACS system functions as a dissimilatory sulfate reduction - methanogenic reactor consisting of two compartments, a shear-loop anaerobic bed (SLAB) unit and an anaerobic plug flow (APF) unit. The functional role of the SLAB unit is not limited to acidogenesis but also sulfidogenic processes, which curtails H2S generation in the APF unit (methanogenic stage). Experimental observations indicated that pH serves a critical role in the cohabitation of acidogenic and sulfidogenic microbes in the SLAB unit. Although acidogenesis was not influenced by pH within the range of 4.5-6.0, it is vital to stabilize the pH of this unit at 5.4 to establish a steady sulfate reduction of above 75%. The highest desulfurization achieved in this compartment was 88% under a hydraulic retention time (HRT) of 4 h. With an average methane productivity of 256 mL g-1 VS, the methanogenic performance of the two-stage domesticated SLACS system shows a 32% methanogenic proficiency higher than that of the one-stage digestion system. Microbial community structure within the system carried out via Next Generation Sequencing (NGS) provided qualitative data on the sludge's sulfidogenic and methanogenic performance.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  16. G-quadruplex structures bind to EZ-Tn5 transposase
    Cree SL, Chua EW, Crowther J, Dobson RCJ, Kennedy MA
    Biochimie, 2020 Aug 14.
    PMID: 32805304 DOI: 10.1016/j.biochi.2020.07.022
    Next generation DNA sequencing and analysis of amplicons spanning the pharmacogene CYP2D6 suggested that the Nextera transposase used for fragmenting and providing sequencing priming sites displayed a targeting bias. This manifested as dramatically lower sequencing coverage at sites in the amplicon that appeared likely to form G-quadruplex structures. Since secondary DNA structures such as G-quadruplexes are abundant in the human genome, and are known to interact with many other proteins, we further investigated these sites of low coverage. Our investigation revealed that G-quadruplex structures are formed in vitro within the CYP2D6 pharmacogene at these sites, and G-quadruplexes can interact with the hyperactive Tn5 transposase (EZ-Tn5) with high affinity. These findings indicate that secondary DNA structures such as G-quadruplexes may represent preferential transposon integration sites and provide additional evidence for the role of G-quadruplex structures in transposition or viral integration processes.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  17. Microbiota composition data for wild and captive bluestreak cleaner wrasse Labroides dimidiatus (Valenciennes, 1839)
    Okomoda VT, Nurul ANA, Danish-Daniel AM, Oladimeji AS, Abol-Munafi AB, Alabi KI, et al.
    Data Brief, 2020 Oct;32:106120.
    PMID: 32817873 DOI: 10.1016/j.dib.2020.106120
    The Labroides dimidiatus is known as the "doctor fish" because of its role in removing parasites and infectious pathogens from the body of other fishes. This important role played both in wild and captive conditions could represent a novel form of parasitic transmission process mediated by the cleaning activity of the fish. Yet, there is a paucity of data on the microflora associated with this fish which is important for tracking disease infection and generally monitoring the health status of the fish. This article, therefore, represents the first dataset for the microbiota composition of wild and captive L. dimidiatus. Wild fish samples and carriage water were gotten in Terengganu Malaysia around the corals of the Karah Island. The captive sample, however, was obtained from well-known ornamental fish suppliers in Terengganu Malaysia. Thereafter, bacteria present on the skin, in the stomach and the aquarium water were enumerated using culture-independent approaches and Next Generation Sequencing (NGS) technology. Data obtained from the three metagenomic libraries using NGS analysis gave 1,426,740 amplicon sequence reads which are composed of 508 operational taxonomic units (OTUs) for wild samples and 3,238,564 valid reads and 828 OTUs for captive samples. All sequence reads were deposited in the GeneBank (Accession numbers SAMN14260247, SAMN14260248, SAMN14260249, SAMN14260250, SAMN14260251, and SAMN14260252). The dataset presented is associated with the research article "16S rDNA-Based Metagenomic Analysis of Microbial Communities Associated with Wild Labroides dimidiatus From Karah Island, Terengganu, Malaysia" [1]. The microbiota data presented in this article can be used to monitor the health and wellbeing of the ornamental fish, especially under captivity, hence preventing possible cross-infection.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  18. Fulltext Complete chloroplast genome sequence of Gigantochloa verticillata (Bambusodae)
    Zhang YY, Fan LL, Zheng FY, Zhao T, Rong JD, Chen LG, et al.
    Mitochondrial DNA B Resour, 2020 Feb 06;5(1):306-307.
    PMID: 33366532 DOI: 10.1080/23802359.2019.1702484
    Gigantochloa verticillata is produced in Mengla and Jinghong, Yunnan Province, China, and cultivated in Hong Kong. Vietnam, Thailand, India, Indonesia, and Malaysia are distributed and cultivated. We determined the complete chloroplast genome sequence for G. verticillata using Illumina sequencing data. The complete chloroplast sequence is 139,489 bp, including large single-copy (LSC) region of 83,062 bp, small single-copy (SSC) region of 12,877 bp, and a pair of invert repeats (IR) regions of 21,775 bp. Plastid genome contain 132 genes, 85 protein-coding genes, 39 tRNA genes, and 8 rRNA genes. Phylogenetic analysis based on 23 chloroplast genomes indicates that G. verticillata is closely related to Dendrocalamus latiflorus in Bambusodae.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  19. Fulltext MinION: A Novel Tool for Predicting Drug Hypersensitivity?
    Chua EW, Ng PY
    Front Pharmacol, 2016;7:156.
    PMID: 27378921 DOI: 10.3389/fphar.2016.00156
    The launch of the MinION Access Program has caused much activity within the scientific community. MinION represents a keenly anticipated, novel addition to the current melange of commercial sequencers. Driven by the nanopore sequencing mechanism that requires minimal sample manipulation, the device is capable of generating long sequence reads in sizes (up to or exceeding 50 kb) that surpass those of all other platforms. One notable advantage of this feature is that long-range haplotypes can be more accurately resolved; such advantage is particularly pertinent to the genotyping of complex loci such as genes encoding the human leukocyte antigens, which are pivotal determinants of drug hypersensitivity. With this timely, albeit brief, review, we set out to examine the applications on which MinION has been tested thus far, the bioinformatics workflow tailored to the unique characteristics of its extended sequence reads, the device's potential utility in the detection of genetic markers for drug hypersensitivity, and how it may eventually evolve to become fit for diagnostic purposes in the clinical setting.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
  20. Fulltext Using Population and Comparative Genomics to Understand the Genetic Basis of Effector-Driven Fungal Pathogen Evolution
    Plissonneau C, Benevenuto J, Mohd-Assaad N, Fouché S, Hartmann FE, Croll D
    Front Plant Sci, 2017;8:119.
    PMID: 28217138 DOI: 10.3389/fpls.2017.00119
    Epidemics caused by fungal plant pathogens pose a major threat to agro-ecosystems and impact global food security. High-throughput sequencing enabled major advances in understanding how pathogens cause disease on crops. Hundreds of fungal genomes are now available and analyzing these genomes highlighted the key role of effector genes in disease. Effectors are small secreted proteins that enhance infection by manipulating host metabolism. Fungal genomes carry 100s of putative effector genes, but the lack of homology among effector genes, even for closely related species, challenges evolutionary and functional analyses. Furthermore, effector genes are often found in rapidly evolving chromosome compartments which are difficult to assemble. We review how population and comparative genomics toolsets can be combined to address these challenges. We highlight studies that associated genome-scale polymorphisms with pathogen lifestyles and adaptation to different environments. We show how genome-wide association studies can be used to identify effectors and other pathogenicity-related genes underlying rapid adaptation. We also discuss how the compartmentalization of fungal genomes into core and accessory regions shapes the evolution of effector genes. We argue that an understanding of genome evolution provides important insight into the trajectory of host-pathogen co-evolution.
    Matched MeSH terms: High-Throughput Nucleotide Sequencing
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