LuxR-Type SCO6993 Negatively Regulates Antibiotic Production at the Transcriptional Stage by Binding to Promoters of Pathway-Specific Regulatory Genes in Streptomyces coelicolor.

SCO6993 (606 amino acids) in Streptomyces coelicolor belongs to the large ATP-binding regulators of the LuxR family regulators having one DNA-binding motif. Our previous findings predicted that SCO6993 may suppress the production of pigmented antibiotics, actinorhodin, and undecylprodigiosin, in S. coelicolor, resulting in the characterization of its properties at the molecular level. SCO6993-disruptant, S. coelicolor ΔSCO6993 produced excess pigments in R2YE plates as early as the third day of culture and showed 9.0-fold and 1.8-fold increased production of actinorhodin and undecylprodigiosin in R2YE broth, respectively, compared with that by the wild strain and S. coelicolor ΔSCO6993/SCO6993+. Real-time polymerase chain reaction analysis showed that the transcription of actA and actII-ORF4 in the actinorhodin biosynthetic gene cluster and that of redD and redQ in the undecylprodigiosin biosynthetic gene cluster were significantly increased by SCO6993-disruptant. Electrophoretic mobility shift assay and DNase footprinting analysis confirmed that SCO6993 protein could bind only to the promoters of pathway-specific transcriptional activator genes, actII-ORF4 and redD, and a specific palindromic sequence is essential for SCO6993 binding. Moreover, SCO6993 bound to two palindromic sequences on its promoter region. These results indicate that SCO6993 suppresses the expression of other biosynthetic genes in the cluster by repressing the transcription of actII-ORF4 and redD and consequently negatively regulating antibiotic production.

SCO6992, a Protein with ß-Glucuronidase Activity, Complements a Mutation at the absR Locus and Promotes Antibiotic Biosynthesis in Streptomyces coelicolor.

Streptomyces coelicolor is a filamentous soil bacterium producing several kinds of antibiotics. S. coelicolor abs8752 is an abs (antibiotic synthesis deficient)-type mutation at the absR locus; it is characterized by an incapacity to produce any of the four antibiotics synthesized by its parental strain J1501. A chromosomal DNA fragment from S. coelicolor J1501, capable of complementing the abs- phenotype of the abs8752 mutant, was cloned and analyzed. DNA sequencing revealed that two complete ORFs (SCO6992 and SCO6993) were present in opposite directions in the clone. Introduction of SCO6992 in the mutant strain resulted in a remarkable increase in the production of two pigmented antibiotics, actinorhodin and undecylprodigiosin, in S. coelicolor J1501 and abs8752. However, introduction of SCO6993 did not show any significant difference compared to the control, suggesting that SCO6992 is primarily involved in stimulating the biosynthesis of antibiotics in S. coelicolor. In silico analysis of SCO6992 (359 aa, 39.5 kDa) revealed that sequences homologous to SCO6992 were all annotated as hypothetical proteins. Although a metalloprotease domain with a conserved metal-binding motif was found in SCO6992, the recombinant rSCO6992 did not show any protease activity. Instead, it showed very strong ß-glucuronidase activity in an API ZYM assay and toward two artificial substrates, p-nitrophenyl-ß-D-glucuronide and AS-BI-ß-D-glucuronide. The binding between rSCO6992 and Zn2+ was confirmed by circular dichroism spectroscopy. We report for the first time that SCO6992 is a novel protein with ß-glucuronidase activity, that has a distinct primary structure and physiological role from those of previously reported ß-glucuronidases.

L-carnitine treatment attenuates renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction.

BACKGROUND/AIMS: Accumulating evidence indicates that L-carnitine (LC) protects against multiorgan damage through its antioxidant properties and preservation of the mitochondria. Little information is available about the effects of LC on renal fibrosis. This study examined whether LC treatment would provide renoprotection in a rat model of unilateral ureteral obstruction (UUO) and in vitro. METHODS: Sprague-Dawley rats that underwent UUO were treated daily with LC for 7 or 14 days. The influence of LC on renal injury caused by UUO was evaluated by histopathology, and analysis of gene expression, oxidative stress, mitochondrial function, programmed cell death, and phosphatidylinositol 3-kinase (PI3K)/ AKT/forkhead box protein O 1a (FoxO1a) signaling. In addition, H2O2-exposed human kidney cells (HK-2) were treated with LC. RESULTS: LC treatment inhibited expression of proinflammatory and profibrotic cytokines, and was followed by a significant attenuation of tubulointerstitial inflammation and fibrosis. The increased oxidative stress caused by UUO was associated with mitochondrial dysfunction and excessive apoptosis and autophagy via PI3K/AKT/FoxO1a-dependent signaling, and this was abrogated by administration of LC. In H2O2-exposed HK-2 cells, LC decreased intracellular production of reactive oxygen species, and suppressed expression of profibrotic cytokines and reduced the number of apoptotic cells. CONCLUSION: LC protects against the progression of tubulointerstitial fibrosis in an obstructed kidney.

Primary chondrosarcoma of the liver: A case report.

BACKGROUND: Primary chondrosarcoma of the liver are extremely rare. Moreover, there are few reports focusing on typical clinical symptoms and imaging characteristics. Therefore, the diagnosis of chondrosarcoma of the liver remains a challenge. CASE SUMMARY: A 59-year-old male was admitted due to a lesion occupying the right liver lobe that was found by physical examination. Magnetic resonance imaging showed a lobular mass with high T2 weighted image and low T1 weighted image with enhanced internal separation and edge in the right liver. He was diagnosed with liver cystadenoma by using magnetic resonance imaging. At 3 mo later, the magnetic resonance scan showed that the mass was enlarged. Laparoscopic liver tumor resection was performed with a pathological diagnosis of liver chondrosarcoma. Then he received a surgical resection for the recurrent lesion. However, intrahepatic and abdominal metastases were found again at 8 mo after the second operation. The patient then received conservative management and is now under follow-up. CONCLUSION: Primary liver chondrosarcoma generally is presented as lobulated and heterogeneous density/signal, cystic, solid masses without calcification with enhanced edge, internal septa and solid part. The imaging features are closely related to pathology, which may be helpful for clinical diagnosis.

[Effects of water-nitrogen combination on dry matter, nitrogen accumulation and yield of winter wheat]. / æ°´æ°®ç»„åˆå¯¹å†¬å°éº¦å¹²ç‰©è´¨åŠæ°®ç´ ç§¯ç´¯å’Œäº§é‡çš„å½±å“.

In two growing seasons of wheat (2015-2017), we conducted a field trial with Taishan 28 in Tai'an Academy of Agricultural Science Feicheng experimental base, Tai'an City, Shandong Province. There were four irrigation levels of 150 (A1), 300 (A2), 450 (A3), and 600 (A4) m3·hm-2, and four nitrogen application levels of 90 (B1), 135 (B2), 180 (B3), and 225 (B4) kg·hm-2. We examined the effects of the combination effects of irrigation and nitrogen on dry matter accumulation and transport, nitrogen accumulation and transport, water consumption and utilization, photosynthetic characteristics, wheat grain yield and yield components of wheat. The results showed that dry matter accumulation, nitrogen accumulation, vegetative organs production, storage and the transportation volume to grains of the dry matter and nitrogen, and dry matter and nitrogen accumulation of grain in the mature stage of wheat all reached the maximum in A3B3 treatment, which were significantly different from other treatments. Under all the nitrogen treatments, soil water consumption in the 60-200 cm soil layer was A3>A4>A2>A1. Water use efficiency and nitrogen use efficiency in A3B3 treatment were higher than that under A3B4, A4B3 and A4B4. The net photosynthetic rate, stomatal conductance and transpiration rate of flag leaves from 7 to 28 days after flowe-ring were all significantly higher in A3B3 treatment, which was conducive to the photosynthetic synthesis of carbohydrates in wheat. The interaction effect of water and nitrogen addition significantly affected grain yield and yield components. Wheat yield was the highest in A3B3 treatment which reached at 9400 kg·hm-2. In conclusion, the treatment with irrigation of 450 m3·hm-2 and nitrogen of 180 kg·hm-2 could significantly improve dry matter and nitrogen accumulation, and promote transportation volume of the dry matter and nitrogen to grain. Compared with the high water and nitrogen treatment, it could effectively increase water use efficiency and nitrogen use efficiency, enhance photosynthetic capacity of flag leaf, produce more carbohydrate, and increase grain yield.

Effects of Increased NADPH Concentration by Metabolic Engineering of the Pentose Phosphate Pathway on Antibiotic Production and Sporulation in Streptomyces lividans TK24.

Most of the biosynthetic pathways for secondary metabolites are influenced by carbon metabolism and supply of cytosolic NADPH. We engineered carbon distribution to the pentose phosphate pathway (PPP) and redesigned the host to produce high levels of NADPH and primary intermediates from the PPP. The main enzymes producing NADPH in the PPP, glucose 6-phosphate dehydrogenase (encoded by zwf1 and zwf2) and 6-phosphogluconate dehydrogenase (encoded by zwf3), were overexpressed with opc encoding a positive allosteric effector essential for Zwf activity in various combinations in Streptomyces lividans TK24. Most S. lividans transformants showed better cell growth and higher concentration of cytosolic NADPH than those of the control, and S. lividans TK24/pWHM3-Z23O2 containing zwf2+zwf3+opc2 showed the highest NADPH concentration but poor sporulation in R2YE medium. S. lividans TK24/pWHM3-Z23O2 in minimal medium showed the maximum growth (6.2 mg/ml) at day 4. Thereafter, a gradual decrease of biomass and a sharp increase of cytosolic NADPH and sedoheptulose 7-phosphate between days 2 and 4 and between days 1 and 3, respectively, were observed. Moreover, S. lividans TK24/pWHM3-Z23O2 produced 0.9 times less actinorhodin but 1.8 times more undecylprodigiosin than the control. These results suggested that the increased NADPH concentration and various intermediates from the PPP specifically triggered undecylprodigiosin biosynthesis that required many precursors and NADPH-dependent reduction reaction. This study is the first report on bespoke metabolic engineering of PPP routes especially suitable for producing secondary metabolites that need diverse primary precursors and NADPH, which is useful information for metabolic engineering in Streptomyces.

Signal Detection of Imipenem Compared to Other Drugs from Korea Adverse Event Reporting System Database.

PURPOSE: To detect signals of adverse drug events after imipenem treatment using the Korea Institute of Drug Safety & Risk Management-Korea adverse event reporting system database (KIDS-KD). MATERIALS AND METHODS: We performed data mining using KIDS-KD, which was constructed using spontaneously reported adverse event (AE) reports between December 1988 and June 2014. We detected signals calculated the proportional reporting ratio, reporting odds ratio, and information component of imipenem. We defined a signal as any AE that satisfied all three indices. The signals were compared with drug labels of nine countries. RESULTS: There were 807582 spontaneous AEs reports in the KIDS-KD. Among those, the number of antibiotics related AEs was 192510; 3382 reports were associated with imipenem. The most common imipenem-associated AE was the drug eruption; 353 times. We calculated the signal by comparing with all other antibiotics and drugs; 58 and 53 signals satisfied the three methods. We compared the drug labelling information of nine countries, including the USA, the UK, Japan, Italy, Switzerland, Germany, France, Canada, and South Korea, and discovered that the following signals were currently not included in drug labels: hypokalemia, cardiac arrest, cardiac failure, Parkinson's syndrome, myocardial infarction, and prostate enlargement. Hypokalemia was an additional signal compared with all other antibiotics, and the other signals were not different compared with all other antibiotics and all other drugs. CONCLUSION: We detected new signals that were not listed on the drug labels of nine countries. However, further pharmacoepidemiologic research is needed to evaluate the causality of these signals.

Proton pump inhibitors and risk of Clostridium difficile infection: a multi-country study using sequence symmetry analysis.

OBJECTIVE: To determine the association between incident proton pump inhibitor (PPI) use and Clostridium difficile infections across multiple countries Method: National data covering the total population in Australia and Korea, the Canadian population over 65 years and a 3 million person random sample data set from Taiwan were assessed, as were data from a worker insurance population and a hospital inpatient/outpatient population in Japan. Sequence symmetry analysis was used to assess the association with oral vancomycin dispensing as the outcome of interest. RESULTS: 54,957 patients were included. Positive associations were observed in Australia; adjusted sequence ratio (ASR) 2.48 (95% CI 1.90, 3.12), Korea ASR 2.15 (95%CI 2.11, 2.19), Canada ASR 1.45 (95% CI 1.16, 1.79), Japan hospital dataset ASR 3.21 (95%CI 2.12, 4.55) and Japan worker insurance dataset ASR 5.40 (95% CI 2.73, 8.75). The pooled result was ASR 2.40 (95%CI 1.88, 3.05) and 3.16 (95%CI 1.95, 5.10) when limited to Japan, Korean and Taiwan. Results did not vary by individual PPI. The temporal analysis showed effects within the first two weeks of PPI initiation. CONCLUSION: Our study confirms the association between PPI initiation and C. difficile infections across countries in the Asia-Pacific region.

Signal Detection of Adverse Drug Reaction of Amoxicillin Using the Korea Adverse Event Reporting System Database.

We conducted pharmacovigilance data mining for a ß-lactam antibiotics, amoxicillin, and compare the adverse events (AEs) with the drug labels of 9 countries including Korea, USA, UK, Japan, Germany, Swiss, Italy, France, and Laos. We used the Korea Adverse Event Reporting System (KAERS) database, a nationwide database of AE reports, between December 1988 and June 2014. Frequentist and Bayesian methods were used to calculate disproportionality distribution of drug-AE pairs. The AE which was detected by all the three indices of proportional reporting ratio (PRR), reporting odds ratio (ROR), and information component (IC) was defined as a signal. The KAERS database contained a total of 807,582 AE reports, among which 1,722 reports were attributed to amoxicillin. Among the 192,510 antibiotics-AE pairs, the number of amoxicillin-AE pairs was 2,913. Among 241 AEs, 52 adverse events were detected as amoxicillin signals. Comparing the drug labels of 9 countries, 12 adverse events including ineffective medicine, bronchitis, rhinitis, sinusitis, dry mouth, gastroesophageal reflux, hypercholesterolemia, gastric carcinoma, abnormal crying, induration, pulmonary carcinoma, and influenza-like symptoms were not listed on any of the labels of nine countries. In conclusion, we detected 12 new signals of amoxicillin which were not listed on the labels of 9 countries. Therefore, it should be followed by signal evaluation including causal association, clinical significance, and preventability.

Utilization patterns of disease-modifying antirheumatic drugs in elderly rheumatoid arthritis patients.

This study was conducted to investigate disease-modifying antirheumatic drug (DMARD) utilization in Korean elderly patients with rheumatoid arthritis (RA). We used data from January 1, 2005 to June 30, 2006 from the Health Insurance Review and Assessment Service claims database. The study subjects were defined as patients aged 65 yr or older with at least two claims with a diagnosis of RA. DMARD use was compared by the patients' age-group, gender, medical service, and geographic divisions. The patterns of DMARD use in mono- and combination therapy were calculated. RA medication use was calculated by the number of defined daily doses (DDD)/1,000 patients/day. A total of 166,388 patients were identified during the study period. DMARD use in RA patients was 12.0%. The proportion of DMARD use was higher in the younger elderly, females, and patients treated in big cities. Hydroxychloroquine was the most commonly used DMARD in monotherapy, and most of the combination therapies prescribed it with methotrexate. DMARD use in elderly RA patients was noticeably low, although drug prescriptions showed an increasing trend during the study period, clinicians may need to pay more attention to elderly RA patients.

Comparison of the safety of seven iodinated contrast media.

We aimed to determine the characteristic adverse events (AEs) of iodinated contrast media (IOCM) and to compare the safety profiles of different IOCM. This study used the database of AEs reports submitted by healthcare professionals from 15 Regional Pharmacovigilance Centers between June 24, 2009 and December 31, 2010 in Korea. All reports of IOCM, including iopromide, iohexol, iopamidol, iomeprol, ioversol, iobitridol and iodixanol, were analyzed. Safety profiles were compared between different IOCM at the system organ level using the proportional reporting ratio (PRR) and 95% confidence interval (95% CI). Among a total of 48,261 reports, 6,524 (13.5%) reports were related to the use of IOCM. Iopromide (45.5%), iohexol (16.9%), iopamidol (14.3%) and iomeprol (10.3%) were identified as frequently reported media. 'Platelet, bleeding & clotting disorders' (PRR, 29.6; 95%CI, 1.9-472.6) and 'urinary system disorders' (PRR, 22.3; 95% CI, 17.1-29.1) were more frequently reported for iodixanol than the other IOCM. In conclusion, the frequency of AEs by organ class was significantly different between individual media. These differences among different IOCM should be considered when selecting a medium among various IOCM and when monitoring patients during and after its use to ensure optimum usage and patient safety.

H2AFX polymorphisms are associated with decreased risk of diffuse large B cell lymphoma in Koreans.

Polymorphisms of the H2A histone family member X (H2AFX) gene have been associated with decreased non-Hodgkin lymphoma (NHL, -417AA) risk and increased breast cancer (1654AG/GG, and -1420GA/AA) risk. We investigated whether H2AFX polymorphisms are associated with the risk of NHL and its subtypes in 573 NHL Korean patients and 721 cancer-free control subjects, using high resolution melting polymerase chain reaction and an automatic sequencer. There was no association between polymorphisms and the risk of overall NHL, all B cell lymphoma, or all T cell lymphoma. However, the -1420 AA genotype was associated with decreased diffuse large B cell lymphoma (DLBCL) risk (OR, 0.65; 95% CI, 0.43-0.97), and there was a trend for allele dose-effect (p-trend=0.03). The -1187 CC genotype was associated with decreased DLBCL risk with borderline significance (OR, 0.70; 95% CI, 0.48-1.02). There was a trend for an allele dose-effect with borderline significance (p-trend=0.06). These results suggest that the -1420 AA genotype of H2AFX may be associated with reduced DLBCL risks in the Korean population.

Characterization of Sgr3394 produced only by the A-factor-producin Streptomyces griseus IFO 13350, not by the A-factor deficient mutant.

Protein D (9.7 kDa) is an extracellular protein detected in the culture broth of A-factor-producing Streptomyces griseus IFO 13350, but not of the A-factor-deficient mutant strain S. griseus HH1. Comparison of the N-terminal amino acid sequence with the genomic sequencing data of S. griseus IFO 13350 identified protein D as Sgr3394, which encodes a putative secretory protein with unknown function. The premature Sgr3394 consisted of 128 amino acids (13.5 kDa), showed 87.5% identity with SACT1DRAFT-0503, from Streptomyces sp. ACT-1, and 68.8% identity with SrosN15-18634, from S. roseosporus NRRL15998, and was confirmed to be matured for secretion by a peptide cleavage between the Ala-38 and Ala-39 bond. RT-PCR analysis of Sgr3394 clearly showed that it can be transcribed in the wild-type strain, but not in the A-factor-deficient strain. However, a gel-mobility shift assay of the promoter region of sgr3394 with A-factor-dependent transcriptional regulator (AdpA) showed that AdpA could not specifically recognize the putative AdpA-binding site (5'-TCCCCCGAAT-3'). All of these data strongly suggest that the expression of sgr3394 is not directly induced by AdpA but is regulated indirectly by an A-factor dependent protein. Introduction of sgr3394 on a high-copy-numbered plasmid (pWHM3-sgr3394) into S. lividans TK21 induced massive production of actinorhodin (blue pigment) and undecylprodigiosin (red pigment). Compared to the control, production of each pigment increased by 6.1 and 2.6 times, respectively, on R2YE agar, and 3.1 and 1.4 times, respectively, in R2YE broth; there was little influence on morphogenesis. In S. coelicolor A3(2)/pWHM3-sgr3394, actinorhodin and undecylprodigiosin productions were enhanced to 1.8 and 1.1 times those observed in the control, respectively, suggesting that overexpression of sgr3394 can stimulate secondary metabolism, especially actinorhodin biosynthesis, in S. lividans and S. coelicolor.

Fas and FasL polymorphisms are not associated with acute myeloid leukemia risk in Koreans.

Fas and Fas ligand (FasL) polymorphisms in the promoter regions influence transcriptional activities. The interaction of these two genes plays a crucial role in apoptotic cell death regulation. They have been associated with esophageal, lung, uterine cervical, and urinary bladder cancers in human. We performed a case-control study to investigate the association between Fas and FasL polymorphisms and acute myeloid leukemia (AML) risk. Fas−1377G>A (rs2234767), −670T>C (rs1800682), and FasL−844T>C (rs763110) polymorphisms in 592 AML patients and 858 healthy controls were genotyped and tested for associations between polymorphisms and AML risk. There were no significant differences in genotypic and haplotypic distributions and gene-gene interaction between patients and controls in the overall analysis (p>0.05). These results suggested that polymorphisms of Fas and FasL genes were not associated with AML risk in the Korean population.

PARP-1 Val762Ala polymorphism is associated with reduced risk of non-Hodgkin lymphoma in Korean males.

BACKGROUND: Poly(ADP-ribose) polymerase-1 (PARP-1) is a nuclear enzyme that plays a role in DNA repair, differentiation, proliferation, and cell death. The polymorphisms of PARP-1 have been associated with the risk of various carcinomas, including breast, lung, and prostate. We investigated whether PARP-1 polymorphisms are associated with the risk of non-Hodgkin lymphoma (NHL). METHODS: Subjects from a Korean population consisting of 573 NHL patients and 721 controls were genotyped for 5 PARP-1 polymorphisms (Asp81Asp, Ala284Ala, Lys352Lys, IVS13+118A>G, and Val762Ala) using High Resolution Melting polymerase chain reaction (PCR) and an automatic sequencer. RESULTS: None of the 5 polymorphisms were associated with overall risk for NHL. However, the Val762Ala polymorphism was associated with reduced risk for NHL in males [odds ratio (OR), 0.62; 95% confidence interval (CI), 0.41-0.93 for CC genotype and OR, 0.84; 95% CI, 0.60-1.16 for TC genotype] with a trend toward a gene dose effect (p for trend, 0.02). The Asp81Asp (p for trend, 0.04) and Lys352Lys (p for trend, 0.03) polymorphisms revealed the same trend. In an association study of PARP-1 haplotypes, the haplotype-ACAAC was associated with decreased risk of NHL in males (OR, 0.75; 95% CI, 0.59-0.94). CONCLUSION: The present data suggest that Val762Ala, Asp81Asp, and Lys352Lys polymorphisms and the haplotype-ACAAC in PARP-1 are associated with reduced risk of NHL in Korean males.