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1.
Article in English | MEDLINE | ID: mdl-31156321

ABSTRACT

This paper presents results of construction and operation of a persistent-mode, liquid-helium-free, small-scale prototype magnet for the development of a tabletop 1.5-T "finger" MRI system for osteoporosis screening. The prototype magnet, composed of 2 MgB2 coils, one superconducting joint, and a persistent-current switch (PCS) built from a portion of one coil, was wound with a one continuous ~80-m long unreacted and monofilament MgB2 wire and then reacted. The test magnet was charged successfully and generated the estimated target field of 1.75 T at 5 K with the proposed PCS operation. During initial persistent-mode, the field was slightly decayed due to the index dissipation of the joint; thereafter it sustained the persistent field of 1.7 T for 35 h. The test results validated the joint resistance of < 1.2 × 10-11 as well as the proposed approach involving the PCS coil circuit model.

2.
Biotechnol Bioeng ; 113(7): 1421-32, 2016 07.
Article in English | MEDLINE | ID: mdl-26724475

ABSTRACT

A fully automatized robotic platform has been established to facilitate high-throughput screening for protein engineering purposes. This platform enables proper monitoring and control of growth conditions in the microtiter plate format to ensure precise enzyme production for the interrogation of enzyme mutant libraries, protein stability tests and multiple assay screenings. The performance of this system has been exemplified for four enzyme classes important for biocatalysis such as Baeyer-Villiger monooxygenase, transaminase, dehalogenase and acylase in the high-throughput screening of various mutant libraries. This allowed the identification of novel enzyme variants in a sophisticated and highly reliable manner. Furthermore, the detailed optimization protocols should enable other researchers to adapt and improve their methods. Biotechnol. Bioeng. 2016;113: 1421-1432. © 2016 Wiley Periodicals, Inc.


Subject(s)
Automation, Laboratory , Enzyme Assays , High-Throughput Screening Assays , Protein Engineering , Robotics/instrumentation , Automation, Laboratory/instrumentation , Automation, Laboratory/methods , Enzyme Assays/instrumentation , Enzyme Assays/methods , Equipment Design , High-Throughput Screening Assays/instrumentation , High-Throughput Screening Assays/methods , Protein Engineering/instrumentation , Protein Engineering/methods , Small Molecule Libraries , Transaminases
3.
Appl Microbiol Biotechnol ; 99(21): 8955-62, 2015 Nov.
Article in English | MEDLINE | ID: mdl-25998660

ABSTRACT

Based on natural selection and the survival of the fittest by evolutionary adaption, a smart high-throughput system was developed to select active haloalkane dehalogenase variants from a large mutant library. Only active enzyme variants can hydrolyse toxic halogenated alkanes to promote growth, whereas inactive mutants starve or die due to the toxic compound. With this powerful tool, huge enzyme mutant libraries can be screened within a few days. The selection is done without any artificial substrates that are hard to synthesize and they also resemble typical ones for haloalkane dehalogenases. Three saturation libraries, with a size of more than 10(6) cells, based on inactive variants of the haloalkane dehalogenases DhaA or DhlA were successfully screened to retrieve active enzymes. The enrichment of the active wild-type enzyme in contrast to the inactive variants was about 340-fold. In addition, this selection approach can be applied for continuous directed evolution experiments for the enrichment of cells expressing adapted haloalkane dehalogenases.


Subject(s)
Alkenes/metabolism , Biological Assay/methods , Escherichia coli/growth & development , Genetic Testing/methods , Hydrocarbons, Halogenated/metabolism , Hydrolases/analysis , Hydrolases/genetics , Alkenes/toxicity , Escherichia coli/genetics , Hydrocarbons, Halogenated/toxicity , Mutant Proteins/analysis , Mutant Proteins/genetics
4.
Article in English | MEDLINE | ID: mdl-25427849

ABSTRACT

OBJECTIVE: To describe an unusual case of canine toxicosis from presumptive sea hare ingestion, its management, and outcome. CASE SUMMARY: A young dog was treated for acute toxicosis after partially ingesting a giant sea hare washed up on a beach. The primary symptoms of generalized tremors, mild ataxia, and hyperesthesia occurred rapidly within 20 minutes of ingestion. Decontamination procedures were performed early and the dog made a full recovery within an approximate period of 6 hours. NEW OR UNIQUE INFORMATION PROVIDED: Sea hare toxicosis should be considered as a potential cause of acute onset of tremors, ataxia, and hyperesthesia in previously healthy dogs living in coastal areas.


Subject(s)
Aplysia , Ataxia/veterinary , Dog Diseases/etiology , Foodborne Diseases/veterinary , Hyperesthesia/veterinary , Tremor/veterinary , Animals , Ataxia/etiology , Dogs , Foodborne Diseases/complications , Foodborne Diseases/etiology , Hyperesthesia/etiology , Tremor/etiology
5.
Trends Biotechnol ; 24(7): 312-7, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16759722

ABSTRACT

Imprint lithography is emerging as an alternative nano-patterning technology to traditional photolithography that permits the fabrication of 2D and 3D structures with <100 nm resolution, patterning and modification of functional materials other than photoresist and is low cost, with operational ease for use in developing bio-devices. Techniques for imprint lithography, categorized as either 'molding and embossing' or 'transfer printing', will be discussed in the context of microarrays for genomics, proteomics and tissue engineering. Specifically, fabrication by nanoimprint lithography (NIL), UV-NIL, step and flash imprint lithography (S-FIL), micromolding by elastomeric stamps and micro- and nano-contact printing will be reviewed.


Subject(s)
Genomics/instrumentation , Nanotechnology/methods , Printing/methods , Proteomics/instrumentation , Tissue Engineering/instrumentation , Animals , Genomics/methods , Humans , Microarray Analysis/instrumentation , Proteomics/methods , Tissue Engineering/methods
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