Efficient Microfluidic Screening Method Using a Fluorescent Immunosensor for Recombinant Protein Secretions.

Microbial secretory protein expression is widely used for biopharmaceutical protein production. However, establishing genetically modified industrial strains that secrete large amounts of a protein of interest is time-consuming. In this study, a simple and versatile high-throughput screening method for protein-secreting bacterial strains is developed. Different genotype variants induced by mutagens are encapsulated in microemulsions and cultured to secrete proteins inside the emulsions. The secreted protein of interest is detected as a fluorescence signal by the fluorescent immunosensor quenchbody (Q-body), and a cell sorter is used to select emulsions containing improved protein-secreting strains based on the fluorescence intensity. The concept of the screening method is demonstrated by culturing Corynebacterium glutamicum in emulsions and detecting the secreted proteins. Finally, productive strains of fibroblast growth factor 9 (FGF9) are screened, and the FGF9 secretion increased threefold compared to that of parent strain. This screening method can be applied to a wide range of proteins by fusing a small detection tag. This is a highly simple process that requires only the addition of a Q-body to the medium and does not require the addition of any substrates or chemical treatments. Furthermore, this method shortens the development period of industrial strains for biopharmaceutical protein production.

Multi-modal brain imaging showing brain damage to the orbitofrontal cortex and left hemisphere, in a case of prolonged hypoglycemia-induced transient hemiplegia followed by persistent encephalopathy.

A 21-year-old left-handed male patient was admitted with a 19-h history of coma after substantial insulin injection for suicide attempt. Although the patient recovered from coma 3 days after injury, he experienced transient hemiplegia followed by permanent brain damage. Electroencephalogram (EEG), brain magnetic resonance imaging (MRI), and brain single-photon emission computed tomography (SPECT) identified the localization of this dysfunction, but consistency between clinical symptoms and brain images changed depending on the course of treatment. Transient hemiplegia corresponded to abnormal waveforms on EEG and decreased cerebral blood flow on SPECT, whereas persistent dysfunctions corresponded to abnormal brain regions on MRI and SPECT.

Production of two types of exopolysaccharide by Novosphingobium rosa.

A bacterium, Ikeda-3, that was isolated from the wastewater of a winery was found to produce floating and precipitating exopolysaccharides (EPSs) when grown aerobically in a medium containing sucrose as the sole carbon source. The concentrations of floating and precipitating EPSs were approximately 2.7 g/l and 0.6 g/l, respectively, in a mini-jar fermentor after incubations of the bacterium at 15 degrees C for 5 d in a synthetic medium containing 1% sucrose, 0.01% CaCO3, 0.05% MgSO4, 0.05% K2HPO4, 0.0001% Na2MoO4, 0.05% peptone and 0.82% bicine. A component analysis of two types of EPS suggested that they represent a novel type of sphingan composed of glucose, rhamnose, mannose and mannuroic acid. Glucoronic acid, which is commonly found in sphingans, was absent. From physiological, biochemical and chemotaxonomic characterization, phylogenetic analysis and DNA-DNA relatedness, Ikeda-3 was identified as Novosphingobium rosa. To our knowledge the production of EPS by N. rosa is reported here for the first time.