Engineering of halohydrin hydrogen-halide-lyase (H-lyase) for efficient L-carnitine production

L-carnitine is a beta-hydroxy carboxylic acid known as a dietary supplement with the effect of fat-burning or anti-fatigue. Although several production processes of L-carnitine are proposed including fermentation, chiral resolution or asymmetric synthesis, there are disadvantages with each from the point of view of yield, side-reactions or productivity. We have engaged in technology development of efficient enzymatic production of various chemicals for years, represented with the world’s first industrialized bioprocess for acrylamide. Leveraging our specialty including proprietary nitrile-related enzymes and associated technologies, we attempted to develop a further efficient production process of L-carnitine (Figure 1). Please click Additional Files below to see the full abstract

Successful treatment of persistent bronchorrhea by gefitinib in a case with Recurrent Bronchioloalveolar Carcinoma: a case report

BACKGROUND: Bronchorrhea is one of late complaints in patients with bronchioloalveolar carcinoma (BAC) and hampers their quality of life. Although an effective treatment for bronchorrhea in these patients has not been established, recently we have treated effectively one case of persistent bronchorrhea associated with clinical recurrent BAC with gefitinib (ZD1839, 'Iressa™'; AstraZeneca Japan; Osaka, Japan). CASE PRESENTATION: A 63-year-old Japanese female had undergone left pneumonectomy with radical lymph node dissection (ND2a) for diffuse type bronchioloalveolar carcinoma originated in left lower lobe. Multiple pulmonary metastases in right lung were found one year after operation. Pulmonary metastatic lesion has grown and she complained of progressive symptoms of massive watery sputum and dyspnea, four years after operation. Although her symptom was getting worse in spite of routine treatment, it completely disappeared within 2 weeks of starting oral gefitinib. Thereafter, she has been symptom-free and shows good partial response on repeat scan after 9 months of oral gefitinib. CONCLUSIONS: The dramatic remission of persistent bronchorrhea by gefitinib in the presented case suggests that gefitinib might be a promising option for bronchioloalveolar carcinoma, particularly in cases with severe bronchorrhea. Although it is not possible to comment on whether the improvement came from tumor cell death itself or suppressive effect of mucin synthesis by the epidermal growth factor receptor-tyrosine kinase inhibitory action

CPP32/Yama/apopain cleaves the catalytic component of DNA-dependent protein kinase in the holoenzyme

AbstractDNA-dependent protein kinase (DNA-PK) is composed of a 460-kDa catalytic component (p460) and a DNA-binding component Ku protein. Immunoblot analysis after treatment of Jurkat cells with anti-Fas antibody demonstrated the cleavage of p460 concomitantly with an increase in CPP32/Yama/apopain activity. Recombinant CPP32/Yama/apopain specifically cleaved p460 in the DNA-PK preparation that had been purified from Raji cells into 230- and 160-kDa polypeptides, the latter of which was detected in anti-Fas-treated Jurkat cells. The regulatory component Ku protein was not significantly affected by CPP32/Yama/apopain. DNA-PK activity was decreased with the disappearance of p460 in the incubation of DNA-PK with CPP32/Yama/apopain. These results suggest that the catalytic component of DNA-PK is one of the target proteins for CPP32/Yama/apopain in Fas-mediated apoptosis

X-ray Spectral Study of the Photoionized Stellar Wind in Vela~X-1

We present results from quantitative modeling and spectral analysis of the high mass X-ray binary Vela X-1 obtained with the Chandra HETGS. The spectra exhibit emission lines from H-like and He-like ions driven by photoionization, as well as fluorescent emission lines from several elements in lower charge states. In order to interpret and make full use of the high-quality data, we have developed a simulator, which calculates the ionization and thermal structure of a stellar wind photoionized by an X-ray source, and performs Monte Carlo simulations of X-ray photons propagating through the wind. The emergent spectra are then computed as a function of the viewing angle accurately accounting for photon transport in three dimensions including dynamics. From comparisons of the observed spectra with the simulation results, we are able to find the ionization structure and the geometrical distribution of material in Vela X-1 that can reproduce the observed spectral line intensities and continuum shapes at different orbital phases remarkably well. It is found that a large fraction of X-ray emission lines from highly ionized ions are formed in the region between the neutron star and the companion star. We also find that the fluorescent X-ray lines must be produced in at least three distinct regions --(1)the extended stellar wind, (2)reflection off the stellar photosphere, and (3)in a distribution of dense material partially covering and possibly trailing the neutron star, which may be associated with an accretion wake. Finally, from detailed analysis of the emission lines, we demonstrate that the stellar wind is affected by X-ray photoionization.Comment: 22 pages, 7 tables, 24 figures, accepted for publication in The Astrophysical Journa

Detection of a Fully-resolved Compton Shoulder of the Iron K-alpha Line in the Chandra X-ray Spectrum of GX 301-2

We report the detection of a fully-resolved, Compton-scattered emission line in the X-ray spectrum of the massive binary GX 301-2 obtained with the High Energy Transmission Grating Spectrometer onboard the Chandra X-ray Observatory. The iron K-alpha fluorescence line complex observed in this system consists of an intense narrow component centered at an energy of E = 6.40 keV and a redward shoulder that extends down to ~6.24 keV, which corresponds to an energy shift of a Compton back-scattered iron K-alpha photon. From detailed Monte Carlo simulations and comparisons with the observed spectra, we are able to directly constrain the physical properties of the scattering medium, including the electron temperature and column density, as well as an estimate for the metal abundance.Comment: 13 pages, 4 figures, 1 table, accepted for publication in ApJ Lette

Light-dependent induction of Edn2 expression and attenuation of retinal pathology by endothelin receptor antagonists in Prominin-1- deficient mice

Retinitis pigmentosa (RP) and macular dystrophy (MD) are prevalent retinal degenerative diseases associated with gradual photoreceptor death. These diseases are often caused by genetic mutations that result in degeneration of the retina postnatally after it has fully developed. The Prominin-1 gene (Prom1) is a causative gene for RP and MD, and Prom1- knockout (KO) mice recapitulate key features of these diseases including light-dependent retinal degeneration and stenosis of retinal blood vessels. The mechanisms underlying progression of such degeneration have remained unknown, however. We here analysed early events associated with retinal degeneration in Prom1-KO mice. We found that photoreceptor cell death and glial cell activation occur between 2 and 3 weeks after birth. High-throughput analysis revealed that expression of the endothelin-2 gene (Edn2) was markedly up-regulated in the Prom1-deficient retina during this period. Expression of Edn2 was also induced by light stimulation in Prom1-KO mice that had been reared in the dark. Finally, treatment with endothelin receptor antagonists attenuated photoreceptor cell death, gliosis, and retinal vessel stenosis in Prom1-KO mice. Our findings suggest that inhibitors of endothelin signalling may delay the progression of RP and MD and therefore warrant further study as potential therapeutic agents for these diseases

Roots-eye view: using microdialysis and microCT to non-destructively map root nutrient depletion and accumulation zones

Improvement in fertiliser use efficiency is a key aspect for achieving sustainable agriculture in order to minimise costs, greenhouse gas emissions and pollution from nutrient runoff. To optimise root architecture for nutrient uptake and efficiency we need to understand what the roots encounter in their environment. Traditional methods of nutrient sampling such as salt extractions can only be done at the end of an experiment, are impractical for sampling locations precisely and give total nutrient values which can overestimate the nutrients available to the roots. In contrast, microdialysis provides a non-invasive, continuous method for sampling available nutrients in the soil. Here for the first time we have used microCT imaging to position microdialysis probes at known distances from the roots and then measured the available nitrate and ammonium. We found that nitrate accumulated close to roots while ammonium was depleted demonstrating that this combination of complementary techniques provides a unique ability to measure root-available nutrients non-destructively and in almost real-time