다중혈관 관상동맥 환자에서 y-문합을 이용하여 양쪽 내흉동맥만을 사용한 우회술의 조기 성적

Studies of steroid modifications catalyzed by microbial whole cells represent a well-established research area in white biotechnology. Still, advances over the last decade in genetic and metabolic engineering, whole-cell biocatalysis in non-conventional media, and process monitoring raised research in this field to a new level. This review summarizes the data on microbial steroid conversion obtained since 2003. The key reactions of structural steroid functionalization by microorganisms are highlighted including sterol side-chain degradation, hydroxylation at various positions of the steroid core, and redox reactions. We also describe methods for enhancement of bioprocess productivity, selectivity of target reactions, and application of microbial transformations for production of valuable pharmaceutical ingredients and precursors. Challenges and prospects of whole-cell biocatalysis applications in steroid industry are discussed.

Microbial steroid transformations: current state and prospects

Dehydrogenases which depend on nicotinamide coenzymes are of increasing interest for the preparation of chiral compounds, either by reduction of a prochiral precursor or by oxidative resolution of their racemate The regeneration of oxidized and reduced nicotinamide cofactors is a very crucial step because the use of these cofactors in stoichiometric amounts is too expensive for application There are several possibilities to regenerate nicotinamide cofactors: established methods such as formate/formate dehydrogenase (FDH) for the regeneration of NADH, recently developed electrochemical methods based on new mediator structures, or the application of gene cloning methods for the construction of “designed” cells by heterologous expression of appropriate genes

https://link.springer.com/content/pdf/10.1007%2F10_2009_55.pdf

Regeneration of nicotinamide coenzymes: principles and applications for the synthesis of chiral compounds.

Activation of the peroxisome proliferator-activated receptor gamma (PPAR-γ) has been proposed as a possible neuroprotective strategy to slow down the progression of early Parkinson's disease (PD). Here we report preclinical data on the use of the PPAR-γ agonist pioglitazone (Actos®; Takeda Pharmaceuticals Ltd.) in a paradigm resembling early PD in nonhuman primates. Rhesus monkeys that were trained to perform a battery of behavioral tests received a single intracarotid arterial injection of 20 ml of saline containing 3 mg of the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Twenty-four hours later the monkeys were assessed using a clinical rating scale, matched accordingly to disability, randomly assigned to one of three groups [placebo (n = 5), 2.5 (n = 6) or 5 (n = 5) mg/kg of pioglitazone] and their treatments started. Three months after daily oral dosing, the animals were necropsied. We observed significant improvements in clinical rating score (P = 0.02) in the animals treated with 5 mg/kg compared to placebo. Behavioral recovery was associated with preservation of nigrostriatal dopaminergic markers, observed as higher tyrosine hydroxylase (TH) putaminal optical density (P = 0.011), higher stereological cell counts of TH-ir (P = 0.02) and vesicular monoamine transporter-2 (VMAT-2)-ir nigral neurons (P = 0.006). Stereological cell counts of Nissl stained nigral neurons confirmed neuroprotection (P = 0.017). Pioglitazone-treated monkeys also showed a dose-dependent modulation of CD68-ir inflammatory cells, that was significantly decreased for 5 mg/kg treated animals compared to placebo (P = 0.018). A separate experiment to assess CSF penetration of pioglitazone revealed that 5 mg/kg p.o. induced consistently higher levels than 2.5 mg/kg and 7.5 mg/kg. p.o. Our results indicate that oral administration of pioglitazone is neuroprotective when administered early after inducing a parkinsonian syndrome in rhesus monkeys and supports the concept that PPAR-γ is a viable target against neurodegeneration.

/pdf/the-ppar-g-agonist-pioglitazone-modulates-inflammation-and-3b0owtl5b3.pdf

The PPAR-γ agonist pioglitazone modulates inflammation and induces neuroprotection in parkinsonian monkeys

Androstenedione production by biotransformation of phytosterols.

High-performance liquid chromatographic method for the determination of pioglitazone in human plasma using ultraviolet detection and its application to a pharmacokinetic study.

A simple bioanalytical assay for determination of montelukast in human plasma: application to a pharmacokinetic study.

The selected standard bacterial strains ( Arthrobacter citreus TISTR 820, Bacillus sphaericus ATCC 13805, Bacilllus stearothermophilus TISTR 329, Corynebacterium aquaticum TISTR 823, Pseudomonas acidovorans TISTR 356, Streptomyces peucetius TISTR 3355) and fungal strains ( Aspergillus niger TISTR 3254, Aspergillus terreus TISTR 3109, Cunninghemella elegans TISTR 3370, Curvularia lunata TISTR 3292, Penicillin siamensis TISTR 1253, Rhizopus arrhizus TISTR 3188) were screened for sterol side-chain cleavage activity using β-sitosterol as a model substrate. In various solubility conditions, the main predicted products, 4-androstene-3,17-dione (AD), 1,4-androstadiene-3,17-dione (ADD) and testosterone (TS) were not obtained in all screening species. Both of Mycobacterium sp. NRRL B-3683 and Mycobacterium sp. NRRL B-3805 were able to convert β-sitosterol to AD and ADD. The principal product from Mycobacterium sp. NRRL B-3683 was AD, meanwhile Mycobacterium sp. NRRL B-3805 was ADD. Structural feature of sterol substrates affected to androstenone production. Compared to all tested sterols, β-sitosterol was higher converted into total androstenones in yield of 75.87 and 83.86% by Mycobacterium sp. NRRL B-3683 and Mycobacterium sp. NRRL B-3805, respectively. Almost equivalent of maximum AD and ADD with total conversion of 81.83% was observed in mixed cultures of both strains.

Biotransformation of various natural sterols to androstenones by Mycobacterium sp. and some steroid-converting microbial strains

In this study, the effects of Terminalia bellerica methanolic extract (0.1, 1, 10, 100 and 500 g/ml) on the mouse immune system were investigated in vitro. Phagocytic activity and lymphocyte proliferation were assayed. The results indicated the effect of the extract (500 g/ml) on the stimulation of macrophage phagocytosis, through the production of superoxide anions and acid phosphatase, with a phagocytic index (PI) value of approximately 1.5 and 1.3, respectively. For the lymphocyte proliferation assay, the extract (500 g/ml) with phytohemagglutinin exhibited maximal activation, with a stimulation index (SI) value of approximately 5.8. With concanavalin A, lipopolysaccharide, and pokeweed mitogen, similar activation (SI 4.5) of lymphocyte proliferation was observed. However, at low concentrations (0.1 g/ml), T. bellerica extract with concanavalin A and pokeweed mitogen caused suppressant activity (SI 0.7). The results suggested that the effect of extract on T-lymphocyte proliferation occurred through the same mechanism as phytohemagglutinin, concanavalin A and B-lymphocyte proliferation through T-cell independent and T-cell dependent mechanisms, in manners similar to lipopolysaccharide and pokeweed mitogen respectively. It might be concluded that the methanolic extract of T. bellerica affected the mouse immune system, specifically both the cellular and humoral immune response in vitro, corresponding with its folklore applications. These results can be further applied to the treatment of human immune mediated diseases.

Effects of Terminalia bellerica Roxb. methanolic extract on mouse immune response in vitro

A simple high-performance liquid chromatographic method for the determination of doxazosin in human plasma was developed and validated. Prazosin was used as internal standard. After extraction twice with ethyl acetate, chromatographic separation of doxazosin in human plasma was carried out using a reversed-phase Apollo C18 column (250 × 4.6 mm, 5 µm) with mobile phase of methanol–acetonitrile–0.04 m disodium hydrogen orthophosphate (22:22:56, v/v/v) adjusted to pH 4.9 with 0.9 m phosphoric acid and quantified by fluorescence detection operated with an excitation wavelength of 246 nm and an emission wavelength of 389 nm. The lower limit of quantification (LLOQ) of this assay was 1 ng/mL using 500 µL human plasma. Linearity was established over the range 1–25 ng/mL (r2 > 0.9994). The intra- and inter-day accuracy ranged from 90.5 to 104.4% and the coefficient of variation were not more than 8.6% for both intra- and inter-day precision, over the range of the calibration curve. The absolute recoveries of doxazosin and prazosin from human plasma were more than 91%. Doxazosin demonstrated acceptable short-term, long-term and freeze–thaw stability in human plasma. The assay has been successfully applied to plasma sample ana-lysis for pharmacokinetic study. Copyright © 2005 John Wiley & Sons, Ltd.

Pattana Sripalakit

Papers

High-performance liquid chromatographic method for the determination of pioglitazone in human plasma using ultraviolet detection and its application to a pharmacokinetic study.

A simple bioanalytical assay for determination of montelukast in human plasma: application to a pharmacokinetic study.

Biotransformation of various natural sterols to androstenones by Mycobacterium sp. and some steroid-converting microbial strains

Effects of Terminalia bellerica Roxb. methanolic extract on mouse immune response in vitro

Validation and pharmacokinetic application of a method for determination of doxazosin in human plasma by high-performance liquid chromatography