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https://www.readbyqxmd.com/read/27906115/established-cell-surface-markers-efficiently-isolate-highly-overlapping-populations-of-skeletal-muscle-satellite-cells-by-fluorescence-activated-cell-sorting
#1
Claire C Maesner, Albert E Almada, Amy J Wagers
BACKGROUND: Fluorescent-activated cell sorting (FACS) has enabled the direct isolation of highly enriched skeletal muscle stem cell, or satellite cell, populations from postnatal tissue. Several distinct surface marker panels containing different positively selecting surface antigens have been used to distinguish muscle satellite cells from other non-myogenic cell types. Because functional and transcriptional heterogeneity is known to exist within the satellite cell population, a direct comparison of results obtained in different laboratories has been complicated by a lack of clarity as to whether commonly utilized surface marker combinations select for distinct or overlapping subsets of the satellite cell pool...
November 8, 2016: Skeletal Muscle
https://www.readbyqxmd.com/read/27906032/infection-of-rhesus-macaques-with-a-pool-of-simian-immunodeficiency-virus-with-the-envelope-genes-from-acute-hiv-1-infections
#2
Kendall C Krebs, Meijuan Tian, Mohammed Asmal, Binhua Ling, Kenneth Nelson, Kenneth Henry, Richard Gibson, Yuejin Li, Weining Han, Robin J Shattock, Ronald S Veazey, Norman Letvin, Eric J Arts, Yong Gao
BACKGROUND: New simian-human immunodeficiency chimeric viruses with an HIV-1 env (SHIVenv) are critical for studies on HIV pathogenesis, vaccine development, and microbicide testing. Macaques are typically exposed to single CCR5-using SHIVenv which in most instances does not reflect the conditions during acute/early HIV infection (AHI) in humans. Instead of individual and serial testing new SHIV constructs, a pool of SHIVenv_B derived from 16 acute HIV-1 infections were constructed using a novel yeast-based SHIV cloning approach and then used to infect macaques...
November 25, 2016: AIDS Research and Therapy
https://www.readbyqxmd.com/read/27905914/development-of-a-gene-synthesis-platform-for-the-efficient-large-scale-production-of-small-genes-encoding-animal-toxins
#3
Ana Filipa Sequeira, Joana L A Brás, Catarina I P D Guerreiro, Renaud Vincentelli, Carlos M G A Fontes
BACKGROUND: Gene synthesis is becoming an important tool in many fields of recombinant DNA technology, including recombinant protein production. De novo gene synthesis is quickly replacing the classical cloning and mutagenesis procedures and allows generating nucleic acids for which no template is available. In addition, when coupled with efficient gene design algorithms that optimize codon usage, it leads to high levels of recombinant protein expression. RESULTS: Here, we describe the development of an optimized gene synthesis platform that was applied to the large scale production of small genes encoding venom peptides...
December 1, 2016: BMC Biotechnology
https://www.readbyqxmd.com/read/27905892/mir-491-regulates-glioma-cells-proliferation-by-targeting-trim28-in-vitro
#4
Zengxin Qi, Shengyong Cai, Jiajun Cai, Lingchao Chen, Yu Yao, Liang Chen, Ying Mao
BACKGROUND: MicroRNAs are significantly involved in tumorigenesis and progression of glioma. However, the critical part they play in glioma have not been fully elaborated. miR-491 and Tripartite motif containing 28 (TRIM28) are reported to aberrantly express in glioblastoma multiforme (GBM). Here, we detected miR-491 and TRIM28 expression and function in glioma cells. METHODS: We analyzed miR-491 expressions in 20 primary human GBM tissues and 6 control brain tissues by qRT-PCR assays and searched for The Cancer Genome Atlas (TCGA) database...
December 1, 2016: BMC Neurology
https://www.readbyqxmd.com/read/27905507/discovery-of-piperonal-converting-oxidase-involved-in-the-metabolism-of-a-botanical-aromatic-aldehyde
#5
Shiori Doi, Yoshiteru Hashimoto, Chiaki Tomita, Takuto Kumano, Michihiko Kobayashi
Piperonal-catabolizing microorganisms were isolated from soil, the one (strain CT39-3) exhibiting the highest activity being identified as Burkholderia sp. The piperonal-converting enzyme involved in the initial step of piperonal metabolism was purified from strain CT39-3. Gene cloning of the enzyme and a homology search revealed that the enzyme belongs to the xanthine oxidase family, which comprises molybdoenzymes containing a molybdopterin cytosine dinucleotide cofactor. We found that the piperonal-converting enzyme acts on piperonal in the presence of O2, leading to formation of piperonylic acid and H2O2...
December 1, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27905279/cloning-of-the-dna-vaccine-candidate-lack-gene-of-leishmania-infantum
#6
Serkan Karaca, Şirin Sahra Ceylan, Süleyman Yazar, Salih Kuk
OBJECTIVE: Leishmaniasis is caused by an obligate intracellular protozoa belonging to Leishmania genus and listed among major tropical diseases by WHO. Because of the high costs, toxicity, and adverse effects of routinely used compounds in the treatment, alternative treatment and vaccine studies are underway. An effective vaccine has not been developed to date. In this study, we aimed to clone one of the most promising DNA vaccine candidates: the homolog-activated C kinase (LACK) gene of Leishmania infantum...
September 2016: Türkiye Parazitolojii Dergisi
https://www.readbyqxmd.com/read/27905065/spatial-measures-of-genetic-heterogeneity-during-carcinogenesis
#7
K Storey, M D Ryser, K Leder, J Foo
In this work we explore the temporal dynamics of spatial heterogeneity during the process of tumorigenesis from healthy tissue. We utilize a spatial stochastic model of mutation accumulation and clonal expansion in a structured tissue to describe this process. Under a two-step tumorigenesis model, we first derive estimates of a non-spatial measure of diversity: Simpson's Index, which is the probability that two individuals sampled at random from the population are identical, in the premalignant population. We next analyze two new measures of spatial population heterogeneity...
November 30, 2016: Bulletin of Mathematical Biology
https://www.readbyqxmd.com/read/27904974/heat-stress-affects-carbohydrate-metabolism-during-cold-induced-sweetening-of-potato-solanum-tuberosum-l
#8
Derek J Herman, Lisa O Knowles, N Richard Knowles
Tolerance to heat stress for retention of low-temperature sweetening-resistant phenotype in potato is conferred by insensitivity of acid invertase activity to cold induction. Heat stress exacerbated cold sweetening (buildup of reducing sugars) of the LTS (low-temperature sweetening)-susceptible potato (Solanum tuberosum L.) cultivars, Ranger Russet and Russet Burbank, and completely abolished the resistance to cold sweetening in the LTS-resistant cultivars/clones, Sage Russet, GemStar Russet, POR06V12-3 and A02138-2...
November 30, 2016: Planta
https://www.readbyqxmd.com/read/27904648/editing-of-the-urease-gene-by-crispr-cas-in-the-diatom-thalassiosira-pseudonana
#9
Amanda Hopes, Vladimir Nekrasov, Sophien Kamoun, Thomas Mock
BACKGROUND: CRISPR-Cas is a recent and powerful addition to the molecular toolbox which allows programmable genome editing. It has been used to modify genes in a wide variety of organisms, but only two alga to date. Here we present a methodology to edit the genome of Thalassiosira pseudonana, a model centric diatom with both ecological significance and high biotechnological potential, using CRISPR-Cas. RESULTS: A single construct was assembled using Golden Gate cloning...
2016: Plant Methods
https://www.readbyqxmd.com/read/27904140/restrictions-in-the-t-cell-repertoire-of-chronic-lymphocytic-leukemia-high-throughput-immunoprofiling-supports-selection-by-shared-antigenic-elements
#10
A Vardi, E Vlachonikola, M Karypidou, E Stalika, V Bikos, K Gemenetzi, C Maramis, A Siorenta, A Anagnostopoulos, S Pospisilova, N Maglaveras, I Chouvarda, K Stamatopoulos, A Hadzidimitriou
Immunoglobulin (IG) gene repertoire restrictions strongly support antigen selection in the pathogenesis of chronic lymphocytic leukemia (CLL). Given the emerging multifarious interactions between CLL and bystander T cells, we sought to determine whether antigen(s) are also selecting T cells in CLL. We performed a large-scale, next-generation sequencing (NGS) study of the T-cell repertoire, focusing on major stereotyped subsets representing CLL subgroups with undisputed antigenic drive, but also included patients carrying non-subset IG rearrangements to seek for T-cell immunogenetic signatures ubiquitous in CLL...
December 1, 2016: Leukemia: Official Journal of the Leukemia Society of America, Leukemia Research Fund, U.K
https://www.readbyqxmd.com/read/27903891/cloning-independent-markerless-gene-editing-in-streptococcus-sanguinis-novel-insights-in-type-iv-pilus-biology
#11
Ishwori Gurung, Jamie-Lee Berry, Alexander M J Hall, Vladimir Pelicic
Streptococcus sanguinis, a naturally competent opportunistic human pathogen, is a Gram-positive workhorse for genomics. It has recently emerged as a model for the study of type IV pili (Tfp)-exceptionally widespread and important prokaryotic filaments. To enhance genetic manipulation of Streptococcus sanguinis, we have developed a cloning-independent methodology, which uses a counterselectable marker and allows sophisticated markerless gene editing in situ We illustrate the utility of this methodology by answering several questions regarding Tfp biology by (i) deleting single or mutiple genes, (ii) altering specific bases in genes of interest, and (iii) engineering genes to encode proteins with appended affinity tags...
November 29, 2016: Nucleic Acids Research
https://www.readbyqxmd.com/read/27903799/cxcr6-mediated-sivagmsab-entry-into-sabaeus-african-green-monkey-lymphocytes-implicates-widespread-use-of-non-ccr5-pathways-in-natural-host-infections
#12
Katherine S Wetzel, Yanjie Yi, Sarah T C Elliott, Dino Romero, Beatrice Jacquelin, Beatrice H Hahn, Michaela Muller-Trutwin, Cristian Apetrei, Ivona Pandrea, Ronald G Collman
: African green monkeys (AGM) and sooty mangabeys (SM) are well-studied natural hosts of SIV, which do not progress to AIDS when infected with their species-specific viruses. SIV natural hosts express very low levels of the canonical entry coreceptor CCR5, and recent studies have shown that CCR5 is dispensable for SIV infection of SM in vivo, and blocking CCR5 does not prevent ex vivo infection of PBMC from SM or vervet AGM. In both hosts, CXCR6 is an efficient entry pathway in vitro...
November 30, 2016: Journal of Virology
https://www.readbyqxmd.com/read/27903649/structural-mechanism-underpinning-cross-reactivity-of-a-cd8-t-cell-clone-that-recognises-a-peptide-derived-from-human-telomerase-reverse-transcriptase
#13
David K Cole, Hugo A van den Berg, Angharad Lloyd, Michael D Crowther, Konrad Beck, Julia Ekeruche-Makinde, John J Miles, Anna M Bulek, Garry Dolton, Andrea J Schauenburg, Aaron Wall, Anna Fuller, Mathew Clement, Bruno Laugel, Pierre J Rizkallah, Linda Wooldridge, Andrew K Sewell
T-cell cross-reactivity is essential for effective immune surveillance, but has also been implicated as a pathway to autoimmunity. Previous studies have demonstrated that T-cell receptors (TCRs) that focus on a minimal motif within the peptide are able to facilitate a high level of T-cell cross-reactivity. However, the structural database shows that most TCRs exhibit less focussed antigen binding involving contact with more peptide residues. To further explore the structural features that allow the clonally expressed TCR to functionally engage with multiple peptide-major histocompatibility complexes (pMHCs), we examined the ILA1 CD8+ T-cell clone that responds to a peptide sequence derived from human telomerase reverse transcriptase (hTERT)...
November 30, 2016: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/27903645/upgrading-short-read-animal-genome-assemblies-to-chromosome-level-using-comparative-genomics-and-a-universal-probe-set
#14
Joana Damas, Rebecca O'Connor, Marta Farré, Vasileios Panagiotis E Lenis, Henry J Martell, Anjali Mandawala, Katie Fowler, Sunitha Joseph, Martin T Swain, Darren K Griffin, Denis M Larkin
Most recent initiatives to sequence and assemble new species' genomes de-novo fail to achieve the ultimate endpoint to produce contigs, each representing one whole chromosome. Even the best-assembled genomes (using contemporary technologies) consist of sub-chromosomal sized scaffolds. To circumvent this problem, we developed a novel approach that combines computational algorithms to merge scaffolds into chromosomal fragments, PCR-based scaffold verification and physical mapping to chromosomes. Multi-genome-alignment-guided probe selection led to the development of a set of universal avian BAC clones that permit rapid anchoring of multiple scaffolds to chromosomes on all avian genomes...
November 30, 2016: Genome Research
https://www.readbyqxmd.com/read/27903284/gp50-as-a-promising-early-diagnostic-antigen-for-taenia-multiceps-infection-in-goats-by-indirect-elisa
#15
Xing Huang, Jing Xu, Yu Wang, Cheng Guo, Lin Chen, Xiaobin Gu, Weimin Lai, Xuerong Peng, Guangyou Yang
BACKGROUND: Coenurosis is caused by coenurus, the metacestode of Taenia multiceps, which mainly parasitizes the brain and spinal cord of cattle, sheep and goats. To date, no widely-approved methods are available to identify early coenurus infection. METHODS: In this study, we identified a full-length cDNA that encodes GP50 (TmGP50) from the transcriptome of T. multiceps, and then cloned and expressed in E. coli. The native proteins in adult stage and coenurus were located via immunofluorescence assays, while the potential of recombinant TmGP50 protein (rTmGP50) for indirect ELISA-based serodiagnostics was assessed using native goat sera...
December 1, 2016: Parasites & Vectors
https://www.readbyqxmd.com/read/27903276/accurate-prediction-of-response-to-endocrine-therapy-in-breast-cancer-patients-current-and-future-biomarkers
#16
REVIEW
Cigdem Selli, J Michael Dixon, Andrew H Sims
Approximately 70% of patients have breast cancers that are oestrogen receptor alpha positive (ER+) and are therefore candidates for endocrine treatment. Many of these patients relapse in the years during or following completion of adjuvant endocrine therapy. Thus, many ER+ cancers have primary resistance or develop resistance to endocrine therapy during treatment. Recent improvements in our understanding of how tumours evolve during treatment with endocrine agents have identified both changes in gene expression and mutational profiles, in the primary cancer as well as in circulating tumour cells...
December 1, 2016: Breast Cancer Research: BCR
https://www.readbyqxmd.com/read/27903223/releasing-of-herpes-simplex-virus-carrying-ngf-in-subarachnoid-space-promotes-the-functional-repair-in-spinal-cord-injured-rats
#17
Zhi-Wei Chen, Hang-Ping Wang, Feng-Mei Yuan, Xiao Zhang, Xiu-Juan Dong, Rui-Shen Xie, Chao Tian, Bing-Shen Li, Zhen-Wu Sun, Long-Hui Zhou, Jian Liu, Tinghua Wang
BACKGROUND: Spinal cord injury is a serious disease which can lead to bad consequence in patients. Gene therapies as an effective strategy, has been developed for the treatment of disease. OBJECTIVE: Here, we explored the effect of NGF release carried by HSV-1 in the injured spinal cord. METHODS: Transgenic recombinant containing NGF was constructed by using clone technology. NGF was firstly constructed into plasimid, then enveloped by HSV-1 vector...
November 20, 2016: Current Gene Therapy
https://www.readbyqxmd.com/read/27902907/thermal-stability-study-of-five-newcastle-disease-attenuated-vaccine-strains
#18
Zineb Boumart, Jihane Hamdi, Samira Daouam, Amal Elarkam, Khalid Omari Tadlaoui, Mehdi El Harrak
Newcastle disease (ND) is a big concern throughout the world because of the devastating losses that can occur with commercial and backyard poultry. The major problem in many countries is the loss of the vaccine's effectiveness due to inadequate use or storage conditions, particularly in hot climates. In the present study, stability of the five, most-used NDV vaccine strains (I-2, LaSota, B1, Clone 30 [C30], and VG-GA) was tested comparatively at different storage temperatures (4 and 37 C for the freeze-dried form and 4, 24, 37, and 45 C for the freeze-dried vaccine reconstituted in diluents)...
December 2016: Avian Diseases
https://www.readbyqxmd.com/read/27902845/cloning-and-heterologous-expression-of-chlorophyll-a-synthase-in-rhodobacter-sphaeroides
#19
Emre M Ipekoğlu, Koray Göçmen, Mehmet T Öz, Muazzez Gürgan, Meral Yücel
Rhodobacter sphaeroides is a purple non-sulfur bacterium which photoheterotrophically produces hydrogen from organic acids under anaerobic conditions. A gene coding for putative chlorophyll a synthase (chlG) from cyanobacterium Prochlorococcus marinus was amplified by nested polymerase chain reaction and cloned into an inducible-expression plasmid which was subsequently transferred to R. sphaeroides for heterologous expression. Induced expression of chlG in R. sphaeroides led to changes in light absorption spectrum within 400-700 nm...
November 30, 2016: Journal of Basic Microbiology
https://www.readbyqxmd.com/read/27902806/orthology-guided-transcriptome-assembly-of-italian-ryegrass-and-meadow-fescue-for-single-nucleotide-polymorphism-discovery
#20
Štěpán Stočes, Tom Ruttink, Jan Bartoš, Bruno Studer, Steven Yates, Zbigniew Zwierzykowski, Michael Abrouk, Isabel Roldán-Ruiz, Tomasz Książczyk, Elodie Rey, Jaroslav Doležel, David Kopecký
Single-nucleotide polymorphisms (SNPs) represent natural DNA sequence variation. They can be used for various applications including the construction of high-density genetic maps, analysis of genetic variability, genome-wide association studies, and map-based cloning. Here we report on transcriptome sequencing in the two forage grasses, meadow fescue ( Huds.) and Italian ryegrass ( Lam.), and identification of various classes of SNPs. Using the Orthology Guided Assembly (OGA) strategy, we assembled and annotated a total of 18,952 and 19,036 transcripts for Italian ryegrass and meadow fescue, respectively...
November 2016: Plant Genome
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