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Berlin Brandenburg

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  • 1
    Text Resource
    Text Resource
    Digital Repository @ Iowa State University
    Description: Titin is a giant muscle protein that spans the distance from the Z line to the M line in sarcomeres of striated muscle myofibrils. One of the functions proposed for titin is that it could act as a molecular scaffold to direct sarcomere assembly. To investigate this role of titin, titin expression was transiently altered in the cultured mouse skeletal muscle cell line C2C12 by either a dominant-negative approach or an antisense mRNA approach. The time course of myofibril assembly was monitored by immunofluorescence labeling with antibodies to actin, alpha-actinin, titin, and myosin from 1 to 5 d after myoblast differentiation was induced. The dominant-negative approach was implemented by transiently transfecting cells with a fusion construct consisting of a cDNA for GFP linked to cDNA encoding a portion of the A band region of titin. The protein encoded by this titin cDNA was expressed in bacteria and shown to interact with myosin in vitro by solid phase binding assays. Transfected myotubes expressing high levels of this fusion protein demonstrated delayed myofibrillar A band assembly compared to non-transfected myotubes and myotubes transfected with GFP vector alone. Assembly of the Z line and thin filament regions of the sarcomere were not affected by overexpression of the A band titin. The antisense approach was implemented by transiently transfecting cells with a fusion construct consisting of a 0.5 kb cDNA for the N-terminal (Z line) region of titin in antisense orientation linked to GFP cDNA. This partial Z line titin cDNA contains the 5' untranslated region and the start codon of the titin message. Expression of this anti-sense titin RNA decreased titin expression and delayed titin organization. Delayed myofibrillar A band assembly was also observed in myotubes transfected with this construct in comparison with non-transfected myotubes and myotubes transfected with the GFP vector alone, while the assembly of alpha-actinin and actin filaments were not affected. Overall, the results from both approaches indicate that titin indeed plays an important role in myofibril assembly. Appropriate titin-myosin interaction seems to be crucial for organizing the myofibrillar A band in embryonic skeletal muscle.
    Keywords: Biochemistry ; Biophysics ; And Molecular Biology ; Biochemistry ; Cell Biology
    Source: Iowa State University
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  • 2
    Language: English
    Description: This paper discusses expressed sequence tag sequencing over grid and cloud computing clusters, specifically for biological applications. In this paper, the authors propose a Web service framework for high-level job scheduling that is developed for scientific applications.
    Keywords: Grid Computing ; Cloud Computing ; Bioinformatics ; Scientific Computing ; High Throughput Computing
    Source: University of North Texas
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  • 3
    Language: English
    Description: This paper discusses an ad hoc query tool for relational databases. Most Web servers hosting biological data limit users to a defined set of search options and output formats that are short of the whole range of options available to users with direct database access. However, to make full use of the wealth of data in the database resource, it is desirable to have an intermediate solution that provides a broad range of flexible query and output options through a Web portal.
    Keywords: Ad Hoc Queries ; Relational Databases
    Source: University of North Texas
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  • 4
    Description: Inflammation and infection of bovine mammary glands, commonly known as mastitis, imposes significant losses each year in the dairy industry worldwide. While several different bacterial species have been identified as causative agents of mastitis, many clinical mastitis cases remain culture negative, even after enrichment for bacterial growth. To understand the basis for this increasingly common phenomenon, the composition of bacterial communities from milk samples was analyzed using culture independent pyrosequencing of amplicons of 16S ribosomal RNA genes (16S rDNA). Comparisons were made of the microbial community composition of culture negative milk samples from mastitic quarters with that of non-mastitic quarters from the same animals. Genomic DNA from culture-negative clinical and healthy quarter sample pairs was isolated, and amplicon libraries were prepared using indexed primers specific to the V1–V2 region of bacterial 16S rRNA genes and sequenced using the Roche 454 GS FLX with titanium chemistry. Evaluation of the taxonomic composition of these samples revealed significant differences in the microbiota in milk from mastitic and healthy quarters. Statistical analysis identified seven bacterial genera that may be mainly responsible for the observed microbial community differences between mastitic and healthy quarters. Collectively, these results provide evidence that cases of culture negative mastitis can be associated with bacterial species that may be present below culture detection thresholds used here. The application of cultureindependent bacterial community profiling represents a powerful approach to understand long-standing questions in animal health and disease.
    Keywords: Milk ; Mastitis ; Bovine Mastitis ; Microbiome ; Sequence Databases ; Dna Isolation ; Bacterial Pathogens ; Mycoplasma ; Large Or Food Animal And Equine Medicine ; Veterinary Infectious Diseases ; Veterinary Microbiology And Immunobiology
    Source: Iowa State University
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  • 5
    Description: Inflammation and infection of bovine mammary glands, commonly known as mastitis, imposes significant losses each year in the dairy industry worldwide. While several different bacterial species have been identified as causative agents of mastitis, many clinical mastitis cases remain culture negative, even after enrichment for bacterial growth. To understand the basis for this increasingly common phenomenon, the composition of bacterial communities from milk samples was analyzed using culture independent pyrosequencing of amplicons of 16S ribosomal RNA genes (16S rDNA). Comparisons were made of the microbial community composition of culture negative milk samples from mastitic quarters with that of non-mastitic quarters from the same animals. Genomic DNA from culture-negative clinical and healthy quarter sample pairs was isolated, and amplicon libraries were prepared using indexed primers specific to the V1–V2 region of bacterial 16S rRNA genes and sequenced using the Roche 454 GS FLX with titanium chemistry. Evaluation of the taxonomic composition of these samples revealed significant differences in the microbiota in milk from mastitic and healthy quarters. Statistical analysis identified seven bacterial genera that may be mainly responsible for the observed microbial community differences between mastitic and healthy quarters. Collectively, these results provide evidence that cases of culture negative mastitis can be associated with bacterial species that may be present below culture detection thresholds used here. The application of culture-independent bacterial community profiling represents a powerful approach to understand long-standing questions in animal health and disease.
    Keywords: Large Or Food Animal And Equine Medicine ; Other Veterinary Medicine ; Veterinary Microbiology And Immunobiology
    Source: Iowa State University
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