Structure-based drug discovery of a corticotropin-releasing hormone receptor 1 antagonist using an X-ray free-electron laser.

Thus far, attempts to develop drugs that target corticotropin-releasing hormone receptor 1 (CRF1R), a drug target in stress-related therapy, have been unsuccessful. Studies have focused on using high-resolution G protein-coupled receptor (GPCR) structures to develop drugs. X-ray free-electron lasers (XFELs), which prevent radiation damage and provide access to high-resolution compositions, have helped accelerate GPCR structural studies. We elucidated the crystal structure of CRF1R complexed with a BMK-I-152 antagonist at 2.75 Å using fixed-target serial femtosecond crystallography. The results revealed that two unique hydrogen bonds are present in the hydrogen bond network, the stalk region forms an alpha helix and the hydrophobic network contains an antagonist binding site. We then developed two antagonists-BMK-C203 and BMK-C205-and determined the CRF1R/BMK-C203 and CRF1R/BMK-C205 complex structures at 2.6 and 2.2 Å, respectively. BMK-C205 exerted significant antidepressant effects in mice and, thus, may be utilized to effectively identify structure-based drugs against CRF1R.

An Angiotensin I converting enzyme polymorphism is associated with clinical phenotype when using differentiation-syndrome to categorize korean bronchial asthma patients.

In this study, genetic analysis was conducted to investigate the association of angiotensin I converting enzyme (ACE) gene polymorphism with clinical phenotype based on differentiation-syndrome of bronchial asthma patients. Differentiation-syndrome is a traditional Korean medicine (TKM) theory in which patients are classified into a Deficiency Syndrome Group (DSG) and an Excess Syndrome Group (ESG) according to their symptomatic classification. For this study, 110 participants were evaluated by pulmonary function test. Among them, 39 patients were excluded because they refused genotyping. Of the remaining patients, 52 with DSG of asthma (DSGA) and 29 with ESG of asthma (ESGA), as determined by the differentiation-syndrome techniques were assessed by genetic analysis. ACE insertion/deletion (I/D) polymorphism analysis was conducted using polymerase chain reaction (PCR). Student's t, chi-square, Fisher and Hardy-Weinberg equilibrium tests were used to compare groups. No significant differences in pulmonary function were observed between DSGA and ESGA. The genotypic frequency of ACE I/D polymorphism was found to differ slightly between DSGA and ESGA (P = .0495). However, there were no significant differences in allelic frequency observed between DSGA and ESGA (P = .7006, OR = 1.1223). Interestingly, the allelic (P = .0043, OR = 3.4545) and genotypic (P = .0126) frequencies of the ACE I/D polymorphism in female patients differed significantly between DSGA and ESGA. Taken together, the results presented here indicate that the symptomatic classification of DSGA and ESGA by differentiation-syndrome in Korean asthma patients could be useful in evaluation of the pathogenesis of bronchial asthma.

Effects of electrochemical reduction reactions on the biodegradation of recalcitrant organic compounds (ROCs) and bacterial community diversity.

Five bacterial species, capable of degrading the recalcitrant organic compounds (ROCs) diethyleneglycol monomethylether (DGMME), 1-amino-2-propanol (APOL), 1-methyl-2- pyrrolidinone (NMP), diethyleneglycol monoethylether (DGMEE), tetraethyleneglycol (TEG), and tetrahydrothiophene 1,1-dioxide (sulfolane), were isolated from an enrichment culture. Cupriavidus sp. catabolized 93.5+/-1.7 mg/l of TEG, 99.3+/-1.2 mg/l of DGMME, 96.1+/-1.6mg/l of APOL, and 99.5+/-0.5mg/l of NMP in 3 days. Acineobacter sp. catabolized 100 mg/l of DGMME, 99.9+/-0.1 mg/l of NMP, and 100 mg/l of DGMEE in 3 days. Pseudomonas sp.3 catabolized 95.7+/-1.2 mg/l of APOL and 99.8+/-0.3 mg/l of NMP. Paracoccus sp. catabolized 98.3+/-0.6 mg/l of DGMME and 98.3+/-1.0 mg/l of DGMEE in 3 days. A maximum 43+/-2.0 mg/l of sulfolane was catabolized by Paracoccus sp. in 3 days. When a mixed culture composed of the five bacterial species was applied to real wastewater containing DGMME, APOL, NMP, DGMEE, or TEG, 92~99% of each individual ROC was catabolized within 3 days. However, at least 9 days were required for the complete mineralization of sulfolane. Bacterial community diversity, analyzed on the basis of the TGGE pattern of 16S rDNA extracted from viable cells, was found to be significantly reduced in a conventional bioreactor after 6 days of incubation. However, biodiversity was maintained after 12 days of incubation in an electrochemical bioreactor. In conclusion, the electrochemical reduction reaction enhanced the diversity of the bacterial community and actively catabolized sulfolane.