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1.
Child Care Health Dev ; 50(3): e13270, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38706405

RESUMEN

INTRODUCTION: Chronic physical disease (CPD) makes life filled with many negative events in adolescents, but not all adolescents experiencing negative life events proceed to develop emotional distress, only those with low emotional distress tolerance (EDT). A valid and reliable scale to measure EDT in CPD adolescents is important for caring for their emotional distress. Therefore, the purpose of this study is to translate the 15-item English version Distress Tolerance Scale (DTS) into a Chinese version and then validate the scale for measuring EDT of adolescents with CPD. METHODS: The 15-item English version DTS was translated into a Chinese version using the translation guidelines for cross-cultural research. Two cohorts of adolescents with CPD were recruited from four hospitals in southern Taiwan, with the first cohort including 124 adolescents with CPD employed to conduct exploratory factor analysis, corrected item-total correlation and reliability testing, while the second cohort, consisting of 238 adolescents with CPD, was utilized to examine confirmatory factor analysis and concurrent validity. RESULTS: The two-factor nine-item Chinese version DTS for Adolescents with CPD (C-DTS-A) was developed. Lower scores of the C-DTS-A were significantly associated with higher diabetes distress, poorer self-management, and worse glycaemic control; their correlation coefficients sequentially were -.40, .17 and -.23. Cronbach's α and the test-retest reliability of the two-factor C-DTS-A ranged from .81 to .87 and from .79 to .89, respectively. CONCLUSION: The two-factor nine-item C-DTS-A with good cross-cultural translation quality was a reliable and valid scale to assess EDT for adolescents with CPD.


Asunto(s)
Comparación Transcultural , Distrés Psicológico , Psicometría , Traducciones , Humanos , Adolescente , Femenino , Masculino , Reproducibilidad de los Resultados , Enfermedad Crónica , Taiwán , Encuestas y Cuestionarios/normas , Estrés Psicológico/diagnóstico , Análisis Factorial , Traducción
2.
JMIR Form Res ; 8: e51076, 2024 Apr 29.
Artículo en Inglés | MEDLINE | ID: mdl-38684083

RESUMEN

BACKGROUND: The adoption of mobile health (mHealth) apps among older adults (>65 years) is rapidly increasing. However, use of such apps has not been fully effective in supporting people with dementia and their caregivers in their daily lives. This is mainly attributed to the heterogeneous quality of mHealth apps, highlighting the need for improved app quality in the development of dementia-related mHealth apps. OBJECTIVE: The aims of this study were (1) to assess the quality and content of mobile apps for dementia management and (2) to investigate the relationship between app quality and download numbers. METHODS: We reviewed dementia-related mHealth apps available in the Google Play Store and Apple App Store in Taiwan. The identified mobile apps were stratified according to a random sampling approach and evaluated by five independent reviewers with sufficient training and proficiency in the field of mHealth and the related health care sector. App quality was scored according to the user version of the Mobile Application Rating Scale. A correlation analysis was then performed between the app quality score and number of app downloads. RESULTS: Among the 17 apps that were evaluated, only one was specifically designed to provide dementia-related education. The mean score for the overall app quality was 3.35 (SD 0.56), with the engagement (mean 3.04, SD 0.82) and information (mean 3.14, SD 0.88) sections of the scale receiving the lowest ratings. Our analyses showed clear differences between the top three- and bottom three-rated apps, particularly in the entertainment and interest subsections of the engagement category where the ratings ranged from 1.4 to 5. The top three apps had a common feature in their interface, which included memory, attention, focus, calculation, and speed-training games, whereas the apps that received lower ratings were found to be deficient in providing adequate information. Although there was a correlation between the number of downloads (5000 or more) and app quality (t15=4.087, P<.001), this may not be a significant determinant of the app's perceived impact. CONCLUSIONS: The quality of dementia-related mHealth apps is highly variable. In particular, our results show that the top three quality apps performed well in terms of engagement and information, and they all received more than 5000 downloads. The findings of this study are limited due to the small sample size and possibility of disregarding exceptional occurrences. Publicly available expert ratings of mobile apps could help people with dementia and their caregivers choose a quality mHealth app.

3.
J Invertebr Pathol ; 200: 107976, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-37541570

RESUMEN

The gut microflora plays an important role in insect development and physiology. The gut bacterial microbiome of the fall armyworm (FAW), Spodoptera frugiperda, in both cornfield and laboratory-reared populations was investigated using a 16S metagenomic approach. The alpha- and beta-diversity of the cornfield FAW populations varied among sampling sites and were higher than those of the laboratory-reared FAW population, indicating that different diets and environments influence the gut bacterial composition. To better understand the interaction between the microbiome and entomopathogenic fungi (EPF), FAWs from organic and conventionally managed corn fields and from the laboratory-reared colony were inoculated with Beauveria bassiana NCHU-153 (Bb-NCHU-153). A longer median lethal time (LT50) was observed in the Bb-NCHU-153-infected cornfield FAW population than in the laboratory-reared FAWs. In terms of the microbiome, three Bb-NCHU-153-infected FAW groups showed different gut bacterial compositions compared to noninfected FAW. Further investigation of the cooccurrence network and linear discriminant analysis (LDA) of effect size (LEfSe) revealed that the enriched bacterial genera, such as Enterococcus, Serratia, Achromobacter, and Tsukamurella, in the gut might play the role of opportunistic pathogens after fungal infection; in contrast, some gut bacteria of Methylobacterium, Marinomonas, Paenochrobactrum, Pseudomonas, Acinetobacter, Delftia, Dietzia, Gordonia, Leucobacter, Paracoccus, and Stenotrophomonas might be probiotics against EPF infection. These results indicated that EPF infection can change the gut bacterial composition and lead to a pathobiome in the FAW and that some bacterial species might protect the FAW from EPF infection. These findings could be applied to the design of pathobiome-inducing biocontrol strategies.


Asunto(s)
Beauveria , Microbioma Gastrointestinal , Animales , Spodoptera , Zea mays , Larva
4.
Front Genet ; 14: 1085631, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36741316

RESUMEN

The entomopathogenic fungus (EPF), Beauveria bassiana, is an important and commonly used EPF for microbial control. However, the role of DNA methylation has not been thoroughly studied. Therefore, the whole genomic DNA methylome of one promising EPF isolate, B. bassiana NCHU-157 (Bb-NCHU-157), was investigated by Oxford Nanopore Technologies (ONT). First, the whole genome of Bb-NCHU-157 was sequenced by next-generation sequencing (NGS) and ONT. The genome of Bb-NCHU-157 contains 16 contigs with 34.19 Mb and 50% GC content, which are composed of 10,848 putative protein-coding genes. Two putative DNA methyltransferases (DNMTs) were found, including Dim-2 and C-5 cytosine-specific DNA methylases. Both DNMTs showed higher expression levels in the mycelium stage than in the conidia stage, indicating that development of DNA methylation in Bb-NCHU-157 might occur in the mycelium stage. The global methylation level of the mycelium stage (5 mC = 4.56%, CG = 3.33%, CHG = 0.74%, CHH = 0.49%) was higher than that of the conidial stage (5 mC = 2.99%, CG = 1.99%, CHG = 0.63%, CHH = 0.37%) in both the gene and transposable element (TE) regions. Furthermore, the TE regions showed higher methylation frequencies than the gene regions, especially for CHH site methylation, suggesting regulation of genomic stabilization during mycelium development. In the gene regions, high methylation frequencies were found around the transcription start site (TSS) and transcription end site (TES). Moreover, CG and CHG methylation mainly occur in the promoter and intergenic regions, while CHH methylation occurs in the TE region. Among the methylated regions, 371, 661, and 756 differentially DNA methylated regions (DMRs) were hypermethylated in the mycelium in CG, CHG, and CHH, while only 13 and 7 DMRs were hypomethylated in the mycelium in CHG, and CHH, respectively. Genes located in the DMR shared the GO terms, DNA binding (GO: 0003677), and sequence-specific DNA binding (GO: 0043565) for hypermethylation in the mycelium, suggesting that methylation might regulate gene expression from the initial process. Evaluation of the DNA methylome in Bb-NCHU-157 by ONT provided new insight into this field. These data will be further validated, and epigenetic regulation during the development of B. bassiana will be explored.

5.
Insects ; 13(8)2022 Aug 09.
Artículo en Inglés | MEDLINE | ID: mdl-36005340

RESUMEN

Nosema ceranae is one of the fungal parasites of Apis mellifera. It causes physical and behavioral effects in honey bees. However, only a few studies have reported on gene expression profiling during A. mellifera infection. In this study, the transcriptome profile of mature spores at each time point of infection (5, 10, and 20 days post-infection, d.p.i.) were investigated. Based on the transcriptome and expression profile analysis, a total of 878, 952, and 981 differentially expressed genes (DEGs) (fold change ≥ 2 or ≤ -2) were identified in N. ceranae spores (NcSp) at 5 d.p.i., 10 d.p.i., and 20 d.p.i., respectively. Moreover, 70 upregulated genes and 340 downregulated genes among common DEGs (so-called common DEGs) and 166 stage-specific genes at each stage of infection were identified. The Gene Ontology (GO) analysis indicated that the DEGs and corresponding common DEGs are involved in the functions of cytosol (GO:0005829), cytoplasm (GO:0005737), and ATP binding (GO:0005524). Furthermore, the pathway analysis found that the DEGs and common DEGs are involved in metabolism, environmental information processing, and organismal systems. Four upregulated common DEGs with higher fold-change values, highly associated with spore proteins and transcription factors, were selected for validation. In addition, the stage-specific genes are highly involved in the mechanism of pre-mRNA splicing according to GO enrichment analysis; thus, three of them showed high expression at each d.p.i. and were also subjected to validation. The relative gene expression levels showed a similar tendency as the transcriptome predictions at different d.p.i., revealing that the gene expression of N. ceranae during infection may be related to the mechanism of gene transcription, protein synthesis, and structural proteins. Our data suggest that the gene expression profiling of N. ceranae at the transcriptomic level could be a reference for the monitoring of nosemosis at the genetic level.

6.
Mitochondrial DNA B Resour ; 7(1): 219-221, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35071762

RESUMEN

The complete mitochondrial genome (mitogenome) of Attacus atlas formosanus (Villiard, 1969) is 15,280 bp in length, with the typical gene content and arrangement usually observed in Insecta. It contains 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and one AT-rich region. The overall nucleotide composition of the mitogenome was 39.8% A, 12.9% C, 7.7% G, and 39.6% T, with an A + T bias of 79.4%. Phylogenetic analyses of 23 species in Saturniidae and 3 species in Bombycidae by Bayesian inference showed that A. atlas formosanus belonged to the Tribe Attacini, closely related to Tribe Saturniini. Besides, A. atlas formosanus is closely related to A. atlas with 99% sequence identity. This result well supported the taxonomic position of Saturniidae and their close relationship with the family Bombycidae.

7.
Sci Rep ; 11(1): 15028, 2021 07 22.
Artículo en Inglés | MEDLINE | ID: mdl-34294840

RESUMEN

Deformed wing virus (DWV) prevalence is high in honey bee (Apis mellifera) populations. The virus infects honey bees through vertical and horizontal transmission, leading to behavioural changes, wing deformity, and early mortality. To better understand the impacts of viral infection in the larval stage of honey bees, artificially reared honey bee larvae were infected with DWV (1.55 × 1010 copies/per larva). No significant mortality occurred in infected honey bee larvae, while the survival rates decreased significantly at the pupal stage. Examination of DWV replication revealed that viral replication began at 2 days post inoculation (d.p.i.), increased dramatically to 4 d.p.i., and then continuously increased in the pupal stage. To better understand the impact of DWV on the larval stage, DWV-infected and control groups were subjected to transcriptomic analysis at 4 d.p.i. Two hundred fifty-five differentially expressed genes (DEGs) (fold change ≥ 2 or ≤ -2) were identified. Of these DEGs, 168 genes were downregulated, and 87 genes were upregulated. Gene Ontology (GO) analysis showed that 141 DEGs (55.3%) were categorized into molecular functions, cellular components and biological processes. One hundred eleven genes (38 upregulated and 73 downregulated) were annotated by KO (KEGG Orthology) pathway mapping and involved metabolic pathways, biosynthesis of secondary metabolites and glycine, serine and threonine metabolism pathways. Validation of DEGs was performed, and the related gene expression levels showed a similar tendency to the DEG predictions at 4 d.p.i.; cell wall integrity and stress response component 1 (wsc1), cuticular protein and myo-inositol 2-dehydrogenase (iolG) were significantly upregulated, and small conductance calcium-activated potassium channel protein (SK) was significantly downregulated at 4 d.p.i. Related gene expression levels at different d.p.i. revealed that these DEGs were significantly regulated from the larval stage to the pupal stage, indicating the potential impacts of gene expression levels from the larval to the pupal stages. Taken together, DWV infection in the honey bee larval stage potentially influences the gene expression levels from larvae to pupae and reduces the survival rate of the pupal stage. This information emphasizes the consequences of DWV prevalence in honey bee larvae for apiculture.


Asunto(s)
Abejas/genética , Abejas/virología , Perfilación de la Expresión Génica , Interacciones Huésped-Patógeno/genética , Virus ARN , Transcriptoma , Enfermedades de los Animales/genética , Enfermedades de los Animales/mortalidad , Enfermedades de los Animales/virología , Animales , Biología Computacional/métodos , Secuenciación de Nucleótidos de Alto Rendimiento , Larva , Tasa de Supervivencia
8.
Arch Insect Biochem Physiol ; 105(4): e21749, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33075172

RESUMEN

Research on gut microbiota of phytophagous insects has shown to be important for the physiological functions of insect hosts; however, little is known about the changes in gut microbiota when they are suffering from environmental stress or pathogen infections. During rearing of Phasmotaenia lanyuhensis (Phasmatodea: Phasmatidae), sluggish locomotion was usually followed by the death of the insect with a symptom of melanization in the front part of the abdomen. Therefore, the abnormal individuals were initially classified into moribund, light- and serious-symptom based on the level of abnormal physiological circumstances and melanization. The gut microbiota of these samples were further investigated by 16S metagenomic sequencing and the differences in bacterial abundance and structure of bacterial community were analyzed. A decrease in microbiota diversity was observed in the diseased P. lanyuhensis, with the abundance of phyla Proteobacteria and Firmicute relatively higher compared to those without symptom. Interestingly, principal component analysis based on the bacterial richness was correlated to the level of melanization symptom in the diseased P. lanyuhensis, suggested the change in bacterial microbiota involved in this abnormal circumstance. However, the factor that caused the initial alternation of microbiota remains to be identified. Additionally, the lack of bacterial diversity (i.e., absence of Meiothermus and Nubsella spp.) in P. lanyuhensis might reduce the fitness for surviving. This report provided the comprehensive microbiota analysis for P. lanyuhensis and concluded that either the relative abundance or the bacterial diversity of microbiota in the insect digestive system may influence the physiological functions of phytophagous insects.


Asunto(s)
Microbioma Gastrointestinal/fisiología , Insectos/microbiología , Animales , Bacterias/clasificación , Bacterias/genética , Metagenoma , ARN Ribosómico 16S , Análisis de Secuencia de ADN
9.
Cancers (Basel) ; 12(8)2020 Jul 28.
Artículo en Inglés | MEDLINE | ID: mdl-32731431

RESUMEN

Early onset breast cancer (EOBC), diagnosed at age ~40 or younger, is associated with a poorer prognosis and higher mortality rate compared to breast cancer diagnosed at age 50 or older. EOBC poses a serious threat to public health and requires in-depth investigation. We studied a cohort comprising 90 Taiwanese female patients, aiming to unravel the underlying mechanisms of EOBC etiopathogenesis. Sequence data generated by whole-exome sequencing (WES) and whole-genome sequencing (WGS) from white blood cell (WBC)-tumor pairs were analyzed to identify somatic missense mutations, copy number variations (CNVs) and germline missense mutations. Similar to regular breast cancer, the key somatic mutation-susceptibility genes of EOBC include TP53 (40% prevalence), PIK3CA (37%), GATA3 (17%) and KMT2C (17%), which are frequently reported in breast cancer; however, the structural protein-coding genes MUC17 (19%), FLG (16%) and NEBL (11%) show a significantly higher prevalence in EOBC. Furthermore, the top 2 genes harboring EOBC germline mutations, MUC16 (19%) and KRT18 (19%), encode structural proteins. Compared to conventional breast cancer, an unexpectedly higher number of EOBC susceptibility genes encode structural proteins. We suspect that mutations in structural proteins may increase physical permeability to environmental hormones and carcinogens and cause breast cancer to occur at a young age.

10.
BMC Genomics ; 20(1): 419, 2019 May 27.
Artículo en Inglés | MEDLINE | ID: mdl-31133070

RESUMEN

BACKGROUND: The golden birdwing butterfly (Troides aeacus formosanus) is a rarely observed species in Taiwan. Recently, a typical symptom of nuclear polyhedrosis was found in reared T. aeacus larvae. From the previous Kimura-2 parameter (K-2-P) analysis based on the nucleotide sequence of three genes in this isolate, polh, lef-8 and lef-9, the underlying virus did not belong to any known nucleopolyhedrovirus (NPV) species. Therefore, this NPV was provisionally named "TraeNPV". To understand this NPV, the nucleotide sequence of the whole TraeNPV genome was determined using next-generation sequencing (NGS) technology. RESULTS: The genome of TraeNPV is 125,477 bp in length with 144 putative open reading frames (ORFs) and its GC content is 40.45%. A phylogenetic analysis based on the 37 baculoviral core genes suggested that TraeNPV is a Group I NPV that is closely related to Autographa californica nucleopolyhedrovirus (AcMNPV). A genome-wide analysis showed that TraeNPV has some different features in its genome compared with other NPVs. Two novel ORFs (Ta75 and Ta139), three truncated ORFs (pcna, he65 and bro) and one duplicated ORF (38.7 K) were found in the TraeNPV genome; moreover, there are fewer homologous regions (hrs) than there are in AcMNPV, which shares eight hrs within the TraeNPV genome. TraeNPV shares similar genomic features with AcMNPV, including the gene content, gene arrangement and gene/genome identity, but TraeNPV lacks 15 homologous ORFs from AcMNPV in its genome, such as ctx, host cell-specific factor 1 (hcf-1), PNK/PNL, vp15, and apsup, which are involved in the auxiliary functions of alphabaculoviruses. CONCLUSIONS: Based on these data, TraeNPV would be clarified as a new NPV species with defective AcMNPV genomic features. The precise relationship between TraeNPV and other closely related NPV species were further investigated. This report could provide comprehensive information on TraeNPV for evolutionary insights into butterfly-infected NPV.


Asunto(s)
Baculoviridae/genética , Mariposas Diurnas/virología , Genoma Viral , Animales , Baculoviridae/clasificación , Baculoviridae/aislamiento & purificación , Mariposas Diurnas/crecimiento & desarrollo , Replicación del ADN , ADN Viral/química , Genes Duplicados , Genes Virales , Genómica , Especificidad del Huésped/genética , Larva/virología , Sistemas de Lectura Abierta , Filogenia , Homología de Secuencia de Ácido Nucleico , Transcripción Genética , Proteínas Estructurales Virales/genética
11.
PLoS One ; 13(6): e0199471, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29920548

RESUMEN

Single cell transcriptome (SCT) analysis provides superior resolution to illustrate tumor cell heterogeneity for clinical implications. We characterized four SCTs of MCF-7 using 143 housekeeping genes (HKGs) as control, of which lactate dehydrogenase B (LDHB) expression is silenced. These SCT libraries mapped to 11,423, 11,486, 10,380, and 11,306 RefSeq genes (UCSC), respectively. High consistency in HKG expression levels across all four SCTs, along with transcriptional silencing of LDHB, was observed, suggesting a high sensitivity and reproducibility of the SCT analysis. Cross-library comparison on expression levels by scatter plotting revealed a linear correlation and an 83-94% overlap in transcript isoforms and expressed genes were also observed. To gain insight of transcriptional diversity among the SCTs, expressed genes were split into consistently expressed (CE) (expressed in all SCTs) and inconsistently expressed (IE) (expressed in some but not all SCTs) genes for further characterization, along with the 142 expressed HKGs as a reference. Distinct transcriptional strengths were found among these groups, with averages of 1,612.0, 88.0 and 1.2 FPKM for HKGs, CE and IE, respectively. Comparison between CE and IE groups further indicated that expressions of CE genes vary more significantly than that of IE genes. Gene Ontology analysis indicated that proteins encoded by CE genes are mainly involved in fundamental intracellular activities, while proteins encoded by IE genes are mainly for extracellular activities, especially acting as receptors or ion channels. The diversified gene expressions, especially for those encoded by IE genes, may contribute to cancer drug resistance.


Asunto(s)
Neoplasias de la Mama/genética , Perfilación de la Expresión Génica/métodos , Proteínas de Neoplasias/genética , Análisis de la Célula Individual/métodos , Neoplasias de la Mama/patología , Resistencia a Antineoplásicos/genética , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Genes Esenciales/genética , Humanos , Células MCF-7
12.
BMC Med Genomics ; 11(Suppl 1): 16, 2018 02 13.
Artículo en Inglés | MEDLINE | ID: mdl-29504912

RESUMEN

BACKGROUND: Cell-free circulating DNA (cfDNA) is becoming a useful biopsy for noninvasive diagnosis of diseases. Microbial sequences in plasma cfDNA may provide important information to improve prognosis and treatment. We have developed a stringent method to identify microbial species via microbial cfDNA in the blood plasma of early-onset breast cancer (EOBC) patients and healthy females. Empirically, microbe-originated sequence reads were identified by mapping non-human PE reads in cfDNA libraries to microbial databases. Those mapped concordantly to unique microbial species were assembled into contigs, which were subsequently aligned to the same databases. Microbial species uniquely aligned were identified and compared across all individuals on MCRPM (Microbial CfDNA Reads Per Million quality PE reads) basis. RESULTS: The predominant microbial cfDNAs in all plasma samples examined are originated from bacteria and these bacteria were limited to only a few genera. Among those, Acinetobacter johnsonii XBB1 and low levels of Mycobacterium spp. were commonly found in all healthy females, but also present in an EOBC patient. Compared to those in healthy counterparts, bacterial species in EOBC patients are more diverse and more likely to present at high levels. Among these three EOBC patients tested, a patient who has record high titer (2,724 MCRPM) of Pseudomonas mendocina together with 8.82 MCRPM of Pannonibacter phragmitetus has passed away; another patient infected by multiple Sphingomonas species remains alive; while the third patient who has similar microbial species (Acinetobacter johnsonii XBB1) commonly seen in normal controls is having a normal life. CONCLUSIONS: Our preliminary data on the profiles of microbial cfDNA sequences suggested that it may have some prognostic value in cancer patients. Validation in larger number of patients is warranted.


Asunto(s)
Biomarcadores/sangre , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Ácidos Nucleicos Libres de Células/sangre , ADN Bacteriano/sangre , Adulto , Edad de Inicio , Neoplasias de la Mama/microbiología , Estudios de Casos y Controles , Ácidos Nucleicos Libres de Células/genética , ADN Bacteriano/genética , Femenino , Humanos , Pronóstico
13.
Sci Rep ; 7: 40767, 2017 01 17.
Artículo en Inglés | MEDLINE | ID: mdl-28094343

RESUMEN

Body fluid DNA sequencing is a powerful noninvasive approach for the diagnosis of genetic defects, infectious agents and diseases. The success relies on the quantity and quality of the DNA samples. However, numerous clinical samples are either at low quantity or of poor quality due to various reasons. To overcome these problems, we have developed T oligo-primed polymerase chain reaction (TOP-PCR) for full-length nonselective amplification of minute quantity of DNA fragments. TOP-PCR adopts homogeneous "half adaptor" (HA), generated by annealing P oligo (carrying a phosphate group at the 5' end) and T oligo (carrying a T-tail at the 3' end), for efficient ligation to target DNA and subsequent PCR amplification primed by the T oligo alone. Using DNA samples from body fluids, we demonstrate that TOP-PCR recovers minute DNA fragments and maintains the DNA size profile, while enhancing the major molecular populations. Our results also showed that TOP-PCR is a superior method for detecting apoptosis and outperforms the method adopted by Illumina for DNA amplification.


Asunto(s)
Líquidos Corporales , Ácidos Nucleicos Libres de Células , Reacción en Cadena de la Polimerasa , Cartilla de ADN , Humanos , Poli T , Reacción en Cadena de la Polimerasa/métodos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
14.
PLoS One ; 10(6): e0130673, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26125617

RESUMEN

The Taiwanese (Formosan) macaque (Macaca cyclopis) is the only nonhuman primate endemic to Taiwan. This primate species is valuable for evolutionary studies and as subjects in medical research. However, only partial fragments of the mitochondrial genome (mitogenome) of this primate species have been sequenced, not mentioning its nuclear genome. We employed next-generation sequencing to generate 2 x 90 bp paired-end reads, followed by reference-assisted de novo assembly with multiple k-mer strategy to characterize the M. cyclopis mitogenome. We compared the assembled mitogenome with that of other macaque species for phylogenetic analysis. Our results show that, the M. cyclopis mitogenome consists of 16,563 nucleotides encoding for 13 protein-coding genes, 2 ribosomal RNAs and 22 transfer RNAs. Phylogenetic analysis indicates that M. cyclopis is most closely related to M. mulatta lasiota (Chinese rhesus macaque), supporting the notion of Asia-continental origin of M. cyclopis proposed in previous studies based on partial mitochondrial sequences. Our work presents a novel approach for assembling a mitogenome that utilizes the capabilities of de novo genome assembly with assistance of a reference genome. The availability of the complete Taiwanese macaque mitogenome will facilitate the study of primate evolution and the characterization of genetic variations for the potential usage of this species as a non-human primate model for medical research.


Asunto(s)
Genoma Mitocondrial/genética , Macaca/genética , Animales , Secuencia de Bases , Evolución Biológica , Variación Genética/genética , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN/métodos , Taiwán
15.
Sci Rep ; 5: 12411, 2015 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-26202883

RESUMEN

Humoral immunity against diverse pathogens is rapidly elicited from natural antibody repertoires of limited complexity. But the organizing principles underlying the antibody repertoires that facilitate this immunity are not well-understood. We used HER2 as a model immunogen and reverse-engineered murine antibody response through constructing an artificial antibody library encoded with rudimentary sequence and structural characteristics learned from high throughput sequencing of antibody variable domains. Antibodies selected in vitro from the phage-displayed synthetic antibody library bound to the model immunogen with high affinity and specificities, which reproduced the specificities of natural antibody responses. We conclude that natural antibody structural repertoires are shaped to allow functional antibodies to be encoded efficiently, within the complexity limit of an individual antibody repertoire, to bind to diverse protein antigens with high specificity and affinity. Phage-displayed synthetic antibody libraries, in conjunction with high-throughput sequencing, can thus be designed to replicate natural antibody responses and to generate novel antibodies against diverse antigens.


Asunto(s)
Reacciones Antígeno-Anticuerpo/inmunología , Inmunidad Innata/inmunología , Receptor ErbB-2/química , Receptor ErbB-2/inmunología , Secuencia de Aminoácidos , Animales , Sitios de Unión , Humanos , Ratones , Datos de Secuencia Molecular , Unión Proteica , Relación Estructura-Actividad
16.
BMC Syst Biol ; 6 Suppl 2: S10, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-23281822

RESUMEN

BACKGROUND: Current next-generation sequencing (NGS) platforms adopt two types of sequencing mechanisms: by synthesis or by ligation. The former is employed by 454 and Solexa systems, while the latter by SOLiD system. Although the pros and cons for each sequencing mechanism have more or less been discussed in a number of occasions, the potential obstacle imposed by palindromic sequences has not yet been addressed. METHODS: To test the effect of the palindromic region on sequencing efficacy, we clonally amplified a paired-end ditag sequence composed of a 24-bp palindromic sequence flanked by a pair of tags from the E. coli genome. We used the near homogeneous fragments produced from MmeI digestion of the amplified clone to generate a sequencing library for SOLiD 5500xl sequencer. RESULTS: Results showed that, traditional ABI sequencers, which adopt sequencing-by-synthesis mechanism, were able to read through the palindromic region. However, SOLiD 5500xl was unable to do so. Instead, the palindromic region was read as miscellaneous random sequences. Moreover, readable tag sequence turned obscure ~2 bp prior to the palindromic region. CONCLUSIONS: Taken together, we demonstrate that SOLiD machines, which employ sequencing-by-ligation mechanism, are unable to read through the palindromic region. On the other hand, sequencing-by-synthesis sequencers had no difficulty in doing so.


Asunto(s)
Secuencias Invertidas Repetidas , Análisis de Secuencia/métodos , Secuencia de Bases , Clonación Molecular , ADN de Cadena Simple/genética , Desoxirribonucleasas de Localización Especificada Tipo II/metabolismo , Escherichia coli/genética , Biblioteca de Genes , Genoma Bacteriano/genética , Datos de Secuencia Molecular
17.
Phys Chem Chem Phys ; 12(36): 10928-32, 2010 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-20657947

RESUMEN

Since the successful fabrication of semiconductor nanowires, various techniques have been developed to contact these nanowires and to probe their intrinsic electrical properties. Although many novel quasi one-dimensional materials such as Pb(1 - x)Mn(x)Se nanoarrays were recently produced, their intrinsic electron transport properties have not been extensively studied so far. In this work, we demonstrate that an ordinary source-drain configuration of field-effect transistors or the two-probe measurement can be applied to the exploration of the intrinsic properties of nanowires. This two-probe measurement approach also works on highly resistive nanowires without an Ohmic contact issue. By using this method, electron transport behavior, resistivity, and carrier concentrations of ZnO, InP, GaP, and Pb(1 - x)Mn(x)Se semiconductor nanowires have been investigated. Due to the tiny cross-section and few conducting channels, a nanomaterial usually reveals an ultra high resistance. This technique demonstrates a two-probe characterization of nanostructures, paving the simplest way toward electrical characterizations of all high-resistance nanomaterials such as deoxyribonucleic acid (DNA), molecules and organics.

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