Engineering Rhodosporidium toruloides for production of 3-hydroxypropionic acid from lignocellulosic hydrolysate.

Microbial production of valuable bioproducts is a promising route towards green and sustainable manufacturing. The oleaginous yeast, Rhodosporidium toruloides, has emerged as an attractive host for the production of biofuels and bioproducts from lignocellulosic hydrolysates. 3-hydroxypropionic acid (3HP) is an attractive platform molecule that can be used to produce a wide range of commodity chemicals. This study focuses on establishing and optimizing the production of 3HP in R. toruloides. As R. toruloides naturally has a high metabolic flux towards malonyl-CoA, we exploited this pathway to produce 3HP. Upon finding the yeast capable of catabolizing 3HP, we then implemented functional genomics and metabolomic analysis to identify the catabolic pathways. Deletion of a putative malonate semialdehyde dehydrogenase gene encoding an oxidative 3HP pathway was found to significantly reduce 3HP degradation. We further explored monocarboxylate transporters to promote 3HP transport and identified a novel 3HP transporter in Aspergillus pseudoterreus by RNA-seq and proteomics. Combining these engineering efforts with media optimization in a fed-batch fermentation resulted in 45.4 g/L 3HP production. This represents one of the highest 3HP titers reported in yeast from lignocellulosic feedstocks. This work establishes R. toruloides as a host for 3HP production from lignocellulosic hydrolysate at high titers, and paves the way for further strain and process optimization towards enabling industrial production of 3HP in the future.

White blood cell counts, insulin resistance, vitamin D levels and sarcopenia in Korean elderly men.

Sarcopenia is a major determinant of frailty, disability and mortality in the elderly. Whether low-grade inflammation, insulin resistance and vitamin D are independently associated with sarcopenia remains unclear. In our study, sarcopenia was defined as an appendicular skeletal muscle mass divided by height squared (ASM/Ht2) that was <2 SD below the normal means for young adults. Insulin resistance was estimated using the homeostasis model assessment of insulin resistance (HOMA-IR) index [(insulin (IU/mL) × fasting glucose (mg/dL)/18)/22.5]. Data of white blood cell counts and serum 25-hydroxyvitamin D (25-(OH)D) levels were collected in the second and third year (2008-2009) of Fourth Korean National Health and Nutrition Examination Survey (KNHANES IV). The results showed that the prevalence of sarcopenia in Korean elderly men aged more than 65 years was 11.2%. ASM/Ht2 were positively associated with vitamin D levels, but negatively associated with white blood cell counts and HOMA-IR by multiple regression analysis. After adjustment for covariables, sarcopenia was associated with the highest quartile of WBC counts (OR = 2.93, 95% CI = 1.21-7.14) and the highest quartile of serum vitamin D levels (OR = 0.38, 95% CI = 0.15-0.95). In conclusion, the study findings suggest that higher WBC counts and lower vitamin D levels are independently associated with the presence of sarcopenia in community-dwelling elderly men. They also provide a basis for further studies of the complex immune-endocrine network in sarcopenia.

Association between Sarcopenic Obesity and Bone Mineral Density in Middle-Aged and Elderly Korean.

OBJECTIVES: To identify the prevalence of osteosarcopenic obesity and to evaluate the association between sarcopenic obesity and bone mineral density in middle-aged and elderly Korean. METHODS: This study included 3,385 males and 4,064 females (age ≥50 years and in the stage of menopause) who participated in the Korean National Health and Nutrition Examination Survey (KNHANES) from 2008 to 2011. The cutoff value for sarcopenia was 7.26 kg/m2 for men and 5.45 kg/m2 for female. The fat mass was >30% for men and 40% for female; these values were used to define obesity. Based on the combination of sarcopenia and obesity status, all subjects were classified as sarcopenic obese (SO), sarcopenic nonobese (SNO), nonsarcopenic obese (NSO), or nonsarcopenic nonobese. RESULTS: The incidence rates of sarcopenia, sarcopenic obesity, and osteosarcopenic obesity were 31.5, 5.1, and 4.1%, respectively. Following adjustment, the ORs for osteoporosis were 8.67 in the SO group (95% CI 4.19-17.94), 3.85 in the SNO group (95% CI 2.61-5.67), and 1.88 in the NSO group (95% CI 0.72-4.91) in men and 2.93 in the SO group (95% CI 1.99-4.32), 1.71 in the SNO group (95% CI 1.31-2.23), and 0.58 in the NSO group (95% CI 0.43-0.78) in women. CONCLUSIONS: Sarcopenic obesity is associated with the development of osteoporosis among the middle-aged and elderly Korean population.

Association between sarcopenia and metabolic syndrome in chronic obstructive pulmonary disease: the Korea National Health and Nutrition Examination Survey (KNHANES) from 2008 to 2011.

AIM: It is not clear whether the restrictive or obstructive pattern of spirometry is associated with metabolic syndrome. We investigated the associations between restrictive and obstructive spirometric patterns and metabolic risk factors using data from the Korea National Health and Nutrition Examination Survey (KNHANES). Additionally, we investigated whether sarcopenia is associated with metabolic syndrome in patients with chronic obstructive pulmonary disease (COPD). METHODS: Using data from KNHANES between 2008 and 2011, we enrolled 8,145 subjects (normal lung function: 6,077, obstructive spirometric pattern: 1,039, restrictive pattern: 1,029) aged ≥40 years who underwent anthropometric measurement, laboratory tests, spirometry and estimation of appendicular muscle mass. Sarcopenia was defined as an appendicular skeletal muscle mass divided by body weight squared <2 SD below the sex-specific mean for the young reference group. RESULTS: Sarcopenia was found in 32.8% of male and 12.2% of female patients with COPD. The odds ratio (OR) of metabolic syndrome for the restrictive spirometric pattern in male was 1.29 (95% confidence interval [CI], 1.02-1.65), and that for obstructive pattern in males was 0.99 (95% CI, 0.79-1.26) after adjustments for covariables (female restrictive pattern (ORs, 1,45; 95% CI, 1.09-1.91) and female obstructive pattern (ORs 0.73; 95% CI, 0.49-1.09). After adjustment for other confounding factors, the risk of metabolic syndrome was higher in sarcopenic male (OR, 1.88; 95% CI, 1.27-2.77) with COPD than in those without sarcopenia. CONCLUSIONS: The restrictive spirometric pattern is associated with metabolic syndrome, and sarcopenia may contribute to the risk of metabolic syndrome in male patients with COPD.

Engineering of a butyraldehyde dehydrogenase of Clostridium saccharoperbutylacetonicum to fit an engineered 1,4-butanediol pathway in Escherichia coli.

1,4-Butanediol (1,4-BDO) is currently produced from succinate via six enzymatic reactions in an engineered Escherichia coli strain. Butyraldehyde dehydrogenase (Bld) and butanol dehydrogenase of Clostridium saccharoperbutylacetonicum were selected based on their activities of catalyzing the final two reactions in the 1,4-BDO pathway. To fit Bld into the non-natural 1,4-BDO pathway, we engineered it through random mutagenesis. Five Bld mutants were then isolated using a colorimetric Schiff's reagent-based method. Subsequent site-directed mutagenesis of Bld generated the two best Bld mutants, L273I and L273T, which produced 1,4-BDO titers fourfold greater than those of wild-type Bld. The enhanced 1,4-BDO titers obtained using L273I and L273T clearly correlated with their enhanced activities, which were caused by amino acid mutations at position 273 of Bld. The highest titer of 1,4-BDO (660 ± 40 mg/L) was obtained in a knock-out E. coli strain [ΔldhA ΔpflB ΔadhE ΔlpdA::K. lpd(E354K) Δmdh ΔarcA gltA(R164L)] coexpressing Bld273T+Bdh.

Renal function is associated with bone mineral density and arterial stiffness in healthy postmenopausal women.

BACKGROUND/AIMS: This study aimed to investigate the effect of renal function on bone mineral density (BMD) and arterial stiffness in postmenopausal women. METHODS: This is a retrospective cross-sectional study. We studied 252 postmenopausal women who visited a health promotion center for a medical checkup. The estimated glomerular filtration rate (eGFR) was calculated using the Cockcroft-Gault (CG) formula and the modification of diet in renal disease (MDRD) formula. Areal BMD measurements were performed using dual-energy X-ray absorptiometry, and arterial stiffness was measured using the brachial-ankle pulse wave velocity (baPWV). RESULTS: The eGFR according to the CG formula was significantly correlated with age, BMI, follicle-stimulating hormone, thyroid-stimulating hormone, high-density lipoprotein cholesterol, baPWV, and BMD at the lumbar spine, femoral neck, and total hip sites. However, the eGFR according to the MDRD formula was significantly correlated with age and baPWV but not with BMD at the lumbar spine, femoral neck, and total hip sites. Decreased renal function (eGFR <60 ml/min/1.73 m(2) according to the CG formula) was independently associated with decreased BMD at the femoral neck site and with increased baPWV (>1,500 cm/s) after adjusting for confounding variables. CONCLUSION: Postmenopausal women with decreased renal function are more likely to have a decreased BMD and greater arterial stiffness.

Comparison of severity predictive rules for hospitalised nursing home-acquired pneumonia in Korea: a retrospective observational study.

BACKGROUND: Nursing home-acquired pneumonia (NHAP) is the leading cause of death among long-term care residents. AIMS: To compare current scoring indices (NHAP model score, Pneumonia Severity Index (PSI), CURB-65 (confusion, urea nitrogen, respiratory rate, blood pressure, age >65 years) and SOAR (systolic blood pressure, oxygenation, age, respiratory rate)) in predicting mortality and admission to the intensive care unit (ICU) in patients with NHAP. METHODS: This retrospective observational study was conducted between July 2008 and June 2011 using data from the Korean Nursing Home Networks. Two hundred and eight nursing home residents were hospitalised with pneumonia in one general hospital. The primary outcome measure was 30-day all-cause mortality. Secondary outcome measures were intensive respiratory or vasopressor support (IRVS), and severe pneumonia (ICU admission or IRVS). RESULTS: PSI class V showed the highest Youden index (0.45), specificity (66.7%), positive predictive value (PPV, 40.0%), negative predictive value (NPV, 91.5%), and area under the curve (AUC, 0.73) for 30-day mortality. For severe pneumonia, PSI class V showed the highest Youden index (0.40), specificity (72.8%), PPV (62.2%), NPV (77.1%), and AUC (0.70). Similarly, PSI class V showed the highest Youden index (0.35), specificity (68.3%), PPV (51.1%), NPV (80.5%), and AUC (0.69) for IRVS. CONCLUSIONS: The PSI has superior discriminatory power in predicting all three clinical outcomes (30-day mortality, severe pneumonia, and IVRS) compared with the NHAP model score, CURB-65 and SOAR.

Engineering Bacillus subtilis for production of 3-hydroxypropanoic acid.

3-Hydroxypropionic acid (3-HP) is a valuable platform chemical that is used as a precursor for several higher value-added chemical products. There is an increased interest in development of cell factories as a means for the synthesis of 3-HP and various other platform chemicals. For more than a decade, concentrated effort has been invested by the scientific community towards developing bio-based approaches for the production of 3-HP using primarily Escherichia coli and Klebsiella pneumoniae as production hosts. These hosts however might not be optimal for applications in e.g., food industry due primarily to endotoxin production and the pathogenic origin of particularly the K. pneumoniae. We have previously demonstrated that the generally recognized as safe organism Bacillus subtilis can be engineered to produce 3-HP using glycerol, an abundant by-product of the biodiesel industry, as substrate. For commercial exploitation, there is a need to substantially increase the titer. In the present study, we optimized the bioprocess conditions and further engineered the B. subtilis 3-HP production strain. Thereby, using glycerol as substrate, we were able to improve 3-HP production in a 1-L bioreactor to a final titer of 22.9 g/L 3-HP.

Evidence of global-scale As, Mo, Sb, and Tl atmospheric pollution in the antarctic snow.

We report the first comprehensive and reliable time series for As, Mo, Sb, and Tl in the snowpack from Dome Fuji in the central East Antarctic Plateau. Our results show significant enrichment of these elements due to either anthropogenic activities or large volcanic eruptions during the past 50 years. With respect to the values reported from 1960 to 1964, we observed the maximum increases in crustal enrichment factors (EFs) for As (a factor of ~15), Mo (~4), Sb (~4), and Tl (~2) during the period between the 1970s and 1990s, reflecting the global dispersion of anthropogenic pollutants of these elements, even to the most remote areas on Earth. Such enrichments are likely related to emissions of trace elements from nonferrous metal smelting and fossil fuel combustion processes in South America, especially in Chile. A drastic decrease in the As concentration and its EF values was observed after the year 2000 in response to the introduction of environmental regulations in the 1990s to reduce As emissions from the copper industry, primarily in Chile. The observed decrease suggests that governmental regulations for pollution control are effective in reducing air pollution at both the regional and global level.

Enhanced percutaneous delivery of recombinant human epidermal growth factor employing nano-liposome system.

We encapsulated recombinant human epidermal growth factor (rhEGF) into nano-liposomes (NLs) system for topical delivery. The rhEGF-loaded NLs were prepared using a high pressure homogenization method. Morphology and overall particle distribution of NLs were investigated using transmission electron microscopy (TEM) and high resolution microscope (CytoViva™). Particle size, zeta (ζ) potential and encapsulation efficiency were measured and the percutaneous delivery of NLs was evaluated using Franz diffusion cells and immunofluorescence confocal laser scanning microscopy (CLSM). The mean particle size, zeta potential and encapsulation efficiency of the NLs were 155.57 ± 2.59 nm, -57.92 ± 4.35 mV and 9.00 ± 0.39%, respectively. TEM and microscopic analysis showed spherical, very even-sized vesicles approximately 150 nm. The skin permeation and localization of rhEGF were enhanced by NLs. CLSM image analysis provided that the NLs enhanced the permeation and localization of rhEGF in rat skin by facilitating entry through pores of skin.

Relationship between bilirubin and C-reactive protein.

BACKGROUND: Epidemiologic studies have shown that serum total bilirubin is inversely related to an increased risk of coronary artery disease. However, the number of studies which have focused on the relationship between bilirubin and C-reactive protein (CRP), which is a marker reflecting chronic vascular inflammation, are limited. METHOD: This cross-sectional study consisted of 2307 Korean adults who visited the Center for Health Promotion for a routine health check-up from January 2009 to December 2009. RESULT: CRP level showed a declining tendency as total bilirubin increased according to total bilirubin quartiles. Negative relations of CRP with both total and direct bilirubin were found after adjustment of age, body mass index, hypertension, diabetes, hypercholesterolemia, cardiovascular disease, taking aspirin, smoking, alcohol drinking and regular exercise and total bilirubin or direct bilirubin. CONCLUSIONS: This study found that elevation of the two types of bilirubin (total and direct) have a relationship with a low serum CRP level among apparently healthy Korean adults. It is hypothesized that a low serum CRP level may be due to the antioxidant and anti-inflammatory effects of bilirubin metabolism.

Monocyte chemoattractant protein-1 is related to metabolic syndrome and homocysteine in subjects without clinically significant atherosclerotic cardiovascular disease.

BACKGROUND: The features of the metabolic syndrome include glucose intolerance, hypertension, dyslipidemia, and central obesity, all of which are risk factors for diabetes and cardiovascular disease. Monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) play a key role in atherosclerosis. We examined the association between chemokines, such as MCP-1 and IL-8, and metabolic syndrome. METHODS: The present study was comprised of 54 men and 126 women. Subjects with cardiovascular disease such as myocardial infarction, TIA and cerebral infarction were excluded. RESULTS: MCP-1 was positively correlated with homeostasis model assessment of insulin resistance, homocysteine, and mean pulse wave velocity, but IL-8 was not. In multiple regression analysis, age, HOMA-IR and homocysteine were found to be an independent factor associated with MCP-1 adjusted by gender, waist, systolic blood pressure, triglyceride, HDL-cholesterol, and hs-CRP. After adjustment for age and gender, mean MCP-1 was higher in subjects with metabolic syndrome and in subject with high blood pressure among the individual components of the metabolic syndrome. CONCLUSION: MCP-1 was associated with a low-grade systemic inflammatory reaction which is often found in the metabolic syndrome.

Differences in the Fatty Acid Profile, Morphology, and Tetraacetylphytosphingosine-Forming Capability Between Wild-Type and Mutant Wickerhamomyces ciferrii.

One tetraacetylphytosphingosine (TAPS)-producing Wickerhamomyces ciferrii mutant was obtained by exposing wild-type W. ciferrii to γ-ray irradiation. The mutant named 736 produced up to 9.1 g/L of TAPS (218.7 mg-TAPS/g-DCW) during batch fermentation in comparison with 1.7 g/L of TAPS (52.2 mg-TAPS/g-DCW) for the wild type. The highest production, 17.7 g/L of TAPS (259.6 mg-TAPS/g-DCW), was obtained during fed-batch fermentation by mutant 736. Fatty acid (FA) analysis revealed an altered cellular FA profile of mutant 736: decrease in C16:0 and C16:1 FA levels, and increase in C18:1 and C18:2 FA levels. Although a significant change in the cellular FA profile was observed, scanning electron micrographs showed that morphology of wild-type and mutant 736 cells was similar. Genetic alteration analysis of eight TAPS biosynthesis-related genes revealed that there are no mutations in these genes in mutant 736; however, mRNA expression analysis indicated 30% higher mRNA expression of TCS10 among the eight genes in mutant 736 than that in the wild-type. Collectively, these results imply that the enhancement of TAPS biosynthesis in mutant 736 may be a consequence of system-level genetic and physiological alterations of a complicated metabolic network. Reverse metabolic engineering based on system-level omics analysis of mutant 736 can make the mutant more suitable for commercial production of TAPS.

Potentially inappropriate prescribing of cardiovascular system and antiplatelet/anticoagulant drugs among elderly patients: a Korean population-based national study.

OBJECTIVES: To investigate the prevalence of potentially inappropriate prescribing (PIP) for cardiovascular system (CVS) and antiplatelet/anticoagulant (AP/AC) drugs among Korean elderly patients, using the Screening Tool of Older Persons' Prescriptions (STOPP) criteria version 2 and to identify the risk factors related to PIP. METHODS: The 2016 National Aged Patient Sample data, comprising National Health Insurance claim records for a random sample of 20% of patients aged ≥ 65 years, were used to calculate PIP prevalence of outpatient prescriptions. For criteria including drug-disease interactions, PIP prevalence per indication was estimated. RESULTS: Among 1,274,148 elderly patients and 27,062,307 outpatient prescription claims, 100,085 patients (7.85%) and 341,664 claims (1.27%) had one or more PIP. The most frequent PIP was "non-steroidal anti-inflammatory drug with concurrent antiplatelet agent (s) without proton-pump inhibitor prophylaxis" in the claim-level (0.97%) and patient-level (6.33%) analyses. "Beta-blocker with bradycardia" (16.47% of claims) and "angiotensin receptor blockers in patients with hyperkalaemia" (23.89% of claims) showed the highest PIP prevalence per indication. Logistic regression analysis revealed that, among the patient and health care provider characteristics, female, older age, more severe comorbidities, polypharmacy, higher level of healthcare organization, and specialty of prescriber were significantly associated with a higher risk of PIP. CONCLUSIONS: Our findings of a high prevalence of PIP for CVS and AP/AC drugs among the elderly suggest that an effective strategy is urgently needed to improve the prescription practices of these drugs.

Increased γ-glutamyltransferase and decreased total bilirubin are associated with metabolic syndrome in Korean postmenopausal women.

BACKGROUND: The objective of this study was to determine if there is an association between serum hepatic markers and the metabolic syndrome in postmenopausal women. METHODS: This study involved 1229 postmenopausal women aged 44-85 years, who visited the Center for Health Promotion for a health check-up. We excluded subjects from the analysis if they had a daily alcohol consumption of more than 1.5 drinks (alcohol consumption ≥20 g/day) or had chronic viral hepatitis. We also excluded subjects who had abnormal hepatic function, as defined by serum aspartate aminotransferase (AST) or alanine aminotransferase (ALT) >100 IU/L, serum γ-glutamyltransferase (GGT) >100 IU/L, or serum total bilirubin concentrations >2 mg/dL. RESULTS: Serum ALT and GGT concentrations increased in proportion to the number of elements of the metabolic syndrome (p<0.01). However, total bilirubin concentrations decreased (p=0.01). After adjusting for age, body mass index, and the presence of fatty liver in the patients with metabolic syndrome, the odds ratios (95% confidence interval) were 1.38 (0.89-2.15) for log (ALT), 1.69 (1.30-2.20) for log (GGT), and 0.53 (0.33-0.86) for log (total bilirubin). CONCLUSIONS: We found that an increase in GGT and a decrease in total bilirubin was associated with metabolic syndrome in postmenopausal women. Hepatic enzymes could be proposed as simple clinical metabolic markers that identify the metabolic syndrome.

Altered subcallosal and posterior cingulate cortex-based functional connectivity during smoking cue and mental simulation processing in smokers.

BACKGROUND: Long-term cigarette smoking induces sensitization of incentive salience and conditioning of contextual cues which involves brain function alteration across multiple regions. Understanding how nicotine affects hub-based functional connectivities involved in affective and cognitive function can help us determine the treatment strategy for nicotine dependence. METHOD: Functional MRI was conducted on 30 smokers and 30 non-smokers while mentally simulating neutral and smoking hand movements. Smoking cue and mental simulation processing-related changes in functional connectivity strengths of the subcallosal and posterior cingulate cortex (SCC and PCC) with major brain network nodes were examined. RESULTS: Compared to non-smokers, smokers showed cue-induced SCC functional connectivities which were enhanced with the intraparietal sulcus and reduced with the medial prefrontal cortex. The PCC activation and functional connectivity enhancements with the anterior insula cortex and rostro-lateral prefrontal cortex was found during smoking mental simulation. The PCC-lateral prefrontal cortex functional connectivity correlated with nicotine dependence severity. CONCLUSION: The present results demonstrate that smokers can be identified by cue-induced SCC functional connectivity strength decline and increment in the default mode and dorsal attention network nodes. However, nicotine dependence was associated with smoking mental simulation-related PCC-lateral prefrontal cortex functional connectivity strength, suggesting that the development of nicotine dependence may depend on the strength of coupling between the default mode network and the central executive network at the cognitive level.

Microbial Production of Retinyl Palmitate and Its Application as a Cosmeceutical.

Chemically synthesized retinyl palmitate has been widely used in the cosmetic and biotechnology industry. In this study, we aimed to demonstrate the microbial production of retinyl palmitate and the benefits of microbial retinyl palmitate in skin physiology. A heterologous retinyl palmitate biosynthesis pathway was reconstructed in metabolically engineered Escherichia coli using synthetic expression modules from Pantoea agglomerans, Salinibacter ruber, and Homo sapiens. High production of retinyl palmitate (69.96 ± 2.64 mg/L) was obtained using a fed-batch fermentation process. Moreover, application of purified microbial retinyl palmitate to human foreskin HS68 fibroblasts led to increased cellular retinoic acid-binding protein 2 (CRABP2) mRNA level [1.7-fold (p = 0.001) at 100 µg/mL], acceleration of cell proliferation, and enhancement of procollagen synthesis [111% (p < 0.05) at 100 µg/mL], strongly indicating an anti-ageing-related effect of this substance. These results would pave the way for large-scale production of retinyl palmitate in microbial systems and represent the first evidence for the application of microbial retinyl palmitate as a cosmeceutical.

Engineering and application of synthetic nar promoter for fine-tuning the expression of metabolic pathway genes in Escherichia coli.

BACKGROUND: Promoters regulate the expression of metabolic pathway genes to control the flux of metabolism. Therefore, fine-tuning of metabolic pathway gene expression requires an applicable promoter system. In this study, a dissolved oxygen-dependent nar promoter was engineered for fine-tuning the expression levels of biosynthetic pathway enzymes in Escherichia coli. To demonstrate the feasibility of using the synthetic nar promoters in production of biochemicals in E. coli, the d-lactate pathway consisting of one enzyme and the 2,3-butanediol (BDO) pathway consisting of three enzymes were investigated. RESULTS: The spacer sequence of 15 bp between the - 35 and - 10 elements of the upstream region of the wild-type nar promoter was randomized, fused to the GFP gene, transduced into E. coli, and screened by flow cytometry. The sorted synthetic nar promoters were divided into three groups according to fluorescence intensity levels: strong, intermediate, and weak. The selected three representative nar promoters of strong, intermediate, and weak intensities were used to control the expression level of the d-lactate and 2,3-BDO biosynthetic pathway enzymes in E. coli. When the ldhD gene encoding d-lactate dehydrogenase was expressed under the control of the strong synthetic nar promoter in fed-batch cultures of E. coli, the d-lactate titers were 105.6 g/L, 34% higher than those using the wild-type promoter (79.0 g/L). When the three 2,3-BDO pathway genes (ilvBN, aldB, and bdh1) were expressed under the control of combinational synthetic nar promoters (strong-weak-strong) in fed-batch cultures of E. coli, the titers of 2,3-BDO were 88.0 g/L, 72% higher than those using the wild-type promoter (51.1 g/L). CONCLUSIONS: The synthetic nar promoters, which were engineered to have strong, intermediate, and weak intensities, were successfully applied to metabolic engineering of d-lactate and 2,3-BDO pathways in E. coli. By controlling expression levels of d-lactate and 2,3-BDO pathway enzymes using the synthetic nar promoters, the production of d-lactate and 2,3-BDO was increased over that using the wild-type promoter by 34 and 72%, respectively. Thus, this synthetic promoter module system will support the improved production of biochemicals and biofuels through fine-tuning of gene expression levels.

Factors influencing on influenza vaccination and its trends of coverage in patients with diabetes in Korea: A population-based cross-sectional study.

BACKGROUND: Influenza infection is a contagious disease and annual influenza vaccination is recommended to the patients with chronic diseases. Although diabetes is an indication for influenza vaccination, the global rate of influenza vaccination is insufficient. Therefore, our study aimed to elucidate influenza vaccination statuses among patients with diabetes and the related factors in Korea. METHODS: A total of 32,268 subjects (4,540 with and 27,728 without diabetes) from the Korea National Health and Nutrition Examination Survey III-VI (2005-2015) were included. Socioeconomic factors and health-related factors were analyses for the relation of influenza vaccination by Student's t-test, the chi-squared test and a multivariate logistic regression analysis. RESULTS: The influenza vaccination coverage rates were 50.0% in the diabetes mellitus (DM) group and 38.2% in the non-DM group. The trends in influenza vaccination rates during KNHANES III-VI were not significant in each group (P trend  =  0.24 in the DM group, 0.30 in the non-DM group). Socioeconomic (older age, female sex, higher family income, and medical aid insurance) and health-related factors (lack of risky alcohol consumption, obesity, and recent health check-ups) were associated with influenza vaccination among patients with DM. CONCLUSIONS: The rate of influenza vaccination among patients with diabetes is insufficient in Korea. More efforts are needed to increase the influenza vaccination rates among vulnerable at-risk populations.

Prevalence and predictors of potentially inappropriate prescribing of central nervous system and psychotropic drugs among elderly patients: A national population study in Korea.

OBJECTIVES: To investigate the prevalence of potentially inappropriate prescribing (PIP) of central nervous system and psychotropic (CNS-PS) drugs to the Korean elderly population, and to identify PIP-associated factors. METHODS: Ambulatory care visits were identified from the 2013 National Aged Patient Sample (HIRA-APS-2013) data, composed of 20% random samples of all enrollees in the universal health security program aged ≥65 years. The CNS-PS section of Screening Tool of Older Person's potentially inappropriate Prescriptions (STOPP) criteria version 2 was used to identify PIP at these visits. RESULTS: A total of 24,427,069 prescription claims records and 1,122,080 patients were included in the study; 10.73% of the claims and 53.64% of the patients satisfied at least one STOPP criterion in the prescription of CNS-PS drugs. The highest prevalence of PIP was observed for the criteria of "first-generation antihistamines" (FGAH), followed by tricyclic antidepressants (TCA) in patients with prostatism and TCA in patients with dementia. The generalized estimating equation logistic regression analysis showed that the PIP of FGAH was significantly associated with polypharmacy (5-9 drugs: odds ratio (OR) 4.965, 95% confidence interval (CI) 4.936-4.994; ≥10 drugs: OR 5.704, 95% CI 5.604-5.807), less severe health conditions (Charlson Comorbidity Index (CCI)=2: OR 0.852, 95% CI 0.842-0.862; CCI=1: OR 0.975, 95% CI 0.964-0.986), prescriptions from clinics (OR>1.0), and outpatient care by general practitioners (OR>1.0). CONCLUSIONS: Appropriate interventions to reduce PIP should be made, especially for the criteria that indicate a high PIP prevalence. Targeted strategies are necessary to modify the risk factors of PIP identified from this study.