Optimization of isoprene production using a metabolically engineered Escherichia Coli

The volatile C5 hydrocarbon, isoprene is an important platform chemical, which has been used in the manufacture of synthetic rubber for tires and also has the potential for various other applications such as elastomers and adhesives. Moreover, isoprene is convertible to biofuel blend stocks such as C10 gasoline, C15 diesel, and jet fuels because of its higher energy content than other biofuels. Although isoprene is currently derived from petroleum, its sustainable supply has been suffered from price fluctuation of crude oil, high refining cost and energy consumption, and low recovery yield of pure isoprene. As an alternative, the biologically produced isoprene (bio-isoprene) has been developed rapidly for the last decade. Bio-isoprene is synthesized from dimethylallyl diphosphate (DMAPP), which is derived from mevalonate (MVA) pathway or the methylerythritol phosphate (MEP) pathway, by isoprene synthase. In this study, metabolic engineering for enhanced production of bio-isoprene was performed by deletion of relevant genes and optimization of culture condition. In comparison of isoprene production between E.coli DH5α and MG1655, lower isoprene production was observed in MG1655. The lower isoprene production in E. coli MG1655 was ascribed to the presence of recA gene which is absent in the DH5α strain. The deletion of recA gene in E.coli MG1655 allows higher isoprene production than E. coli DH5α. Moreover, the optimized expression of isoprene synthesis pathway with 0.03mM IPTG induction enhanced the isoprene production up to 2,850 mg/L. Overall, isoprene production through the optimization was improved by 28.5-fold compared to the initial production of MG1655 strain. Please click Additional Files below to see the full abstract

Sorbus alnifolia protects dopaminergic neurodegeneration in Caenorhabditis elegans

Context: The twigs of Sorbus alnifolia (Sieb. et Zucc.) K. Koch (Rosaceae) have been used to treat neuro- logical disorders as a traditional medicine in Korea. However, there are limited data describing the efficacy of S. alnifolia in Parkinson’s disease (PD). Objective: This study was conducted to identify the protective effects of the methanol extracts of S. alnifolia (MESA) on the dopaminergic (DA) neurodegeneration in Caenorhabditis elegans. Materials and methods: To test the neuroprotective action of MESA, viability assay was performed after 48 h exposure to 1-methyl-4-phenylpyridine (MMPþ) in PC12 cells and C. elegans (400 lM and 2 mM of MMPþ, respectively). Fluorescence intensity was quantified using transgenic mutants such as BZ555 (Pdat-1::GFP) and and UA57 (Pdat-1::GFP and Pdat-1::CAT-2) to determine MESA’s effects on DA neurode- generation in C. elegans. Aggregation of a-synuclein was observed using NL5901 strain (unc-54p::a- synuclein::YFP). MESA’s protective effects on the DA neuronal functions were examined by food-sensing assay. Lifespan assay was conducted to test the effects of MESA on the longevity. Results: MESA restored MPPþ-induced loss of viability in both PC12 cells and C. elegans (85.8% and 54.9%, respectively). In C. elegans, MESA provided protection against chemically and genetically-induced DA neurodegeneration, respectively. Moreover, food-sensing functions were increased 58.4% by MESA in the DA neuron degraded worms. MESA also prolonged the average lifespan by 25.6%. However, MESA failed to alter a-synuclein aggregation. Discussion and conclusions: These results revealed that MESA protects DA neurodegeneration and recov- ers diminished DA neuronal functions, thereby can be a valuable candidate for the treatment of PD

Comparison of CT and 18F-FDG PET for Detecting Peritoneal Metastasis on the Preoperative Evaluation for Gastric Carcinoma

OBJECTIVE: The aim of our study was to compare the accuracy of CT and (18)FFDG PET for detecting peritoneal metastasis in patients with gastric carcinoma. MATERIALS AND METHODS: One-hundred-twelve patients who underwent a histologic confirmative exam or treatment (laparotomy, n = 107; diagnostic laparoscopy, n = 4; peritoneal washing cytology, n = 1) were retrospectively enrolled. All the patients underwent CT and (18)F-FDG PET scanning for their preoperative evaluation. The sensitivities, specificities and accuracies of CT and (18)FFDG PET imaging for the detection of peritoneal metastasis were calculated and then compared using Fisher's exact probability test (p < 0.05), on the basis of the original preoperative reports. In addition, two board-certified radiologists and two board-certified nuclear medicine physicians independently reviewed the CT and PET scans, respectively. A receiver-operating characteristic curve analysis was performed to compare the diagnostic performance of CT and (18)F-FDG PET imaging for detecting peritoneal metastasis. RESULTS: Based on the original preoperative reports, CT and (18)F-FDG PET showed sensitivities of 76.5% and 35.3% (p = 0.037), specificities of 91.6% and 98.9% (p = 0.035), respectively, and equal accuracies of 89.3% (p = 1.0). The receptor operating characteristics curve analysis showed a significantly higher diagnostic performance for CT (Az = 0.878) than for PET (Az = 0.686) (p = 0.004). The interobserver agreement for detecting peritoneal metastasis was good (kappa value = 0.684) for CT and moderate (kappa value = 0.460) for PET. CONCLUSION: For the detection of peritoneal metastasis, CT was more sensitive and showed a higher diagnostic performance than PET, although CT had a relatively lower specificity than did PET.ope

Impact of multivessel versus single-vessel disease on the association between low diastolic blood pressure and mortality after acute myocardial infarction with revascularization

Background: Previous studies demonstrated a J-shaped relationship between low diastolic blood pressure (DBP) and adverse clinical outcomes in patients with acute myocardial infarction (AMI) that was sensitive to revascularization. Hypothesized herein, was that this relationship differs between patients with multivessel disease (MVD) and those with single-vessel disease due to differing degrees of myocardial ischemic burden. Methods: Among 9,983 AMI patients from the Korea Acute Myocardial Infarction Registry database who underwent percutaneous coronary intervention and were followed up for a median duration of 3.2 years, average on-treatment DBP was calculated at admission, discharge, and every scheduled visit and divided into these parameters: &lt; 70 mmHg, 70–74 mmHg, 75–79 mmHg, and ≥ 80 mmHg. The relationship between average on-treatment DBP and clinical outcomes including all-cause death, cardiovascular (CV) death, non-CV death, and hospitalization for heart failure was analyzed using the Cox regression models adjusted for clinical covariates. Results: In patients with MVD, all-cause death (hazard ratio [HR]: 1.47; 95% confidence interval [CI]: 1.06–2.04, p = 0.012) and CV death (HR: 1.59; 95% CI: 1.02–2.46, p = 0.027) were significantly increased in patients with a DBP &lt; 70 mmHg, showing a J-shaped relationship. However, these findings were not significant for single-vessel disease. On a sensitivity analysis excluding subjects with a baseline SBP &lt; 120 mmHg, an increased risk of a low DBP &lt; 70 mmHg remained in MVD. Conclusions: The J-shaped relationship between low DBP and adverse clinical outcomes in AMI patients who underwent revascularization persisted in MVD, which has a high ischemic burden. These high-risk patients require cautious treatment

A Nuclear Localization of the Infectious Haematopoietic Necrosis Virus NV Protein Is Necessary for Optimal Viral Growth

The nonvirion (NV) protein of infectious hematopoietic necrosis virus (IHNV) has been previously reported to be essential for efficient growth and pathogenicity of IHNV. However, little is known about the mechanism by which the NV supports the viral growth. In this study, cellular localization of NV and its role in IHNV growth in host cells was investigated. Through transient transfection in RTG-2 cells of NV fused to green fluorescent protein (GFP), a nuclear localization of NV was demonstrated. Deletion analyses showed that the 32EGDL35 residues were essential for nuclear localization of NV protein, and fusion of these 4 amino acids to GFP directed its transport to the nucleus. We generated a recombinant IHNV, rIHNV-NV-ΔEGDL in which the 32EGDL35 was deleted from the NV. rIHNVs with wild-type NV (rIHNV-NV) or with the NV gene replaced with GFP (rIHNV-ΔNV-GFP) were used as controls. RTG-2 cells infected with rIHNV-ΔNV-GFP and rIHNV-NV-ΔEGDL yielded 12- and 5-fold less infectious virion, respectively, than wild type rIHNV-infected cells at 48 h post-infection (p.i.). While treatment with poly I∶C at 24 h p.i. did not inhibit replication of wild-type rIHNVs, replication rates of rIHNV-ΔNV-GFP and rIHNV-NV-ΔEGDL were inhibited by poly I∶C. In addition, both rIHNV-ΔNV and rIHNV-NV-ΔEGDL induced higher levels of expressions of both IFN1 and Mx1 than wild-type rIHNV. These data suggest that the IHNV NV may support the growth of IHNV through inhibition of the INF system and the amino acid residues of 32EGDL35 responsible for nuclear localization are important for the inhibitory activity of NV

Sorbus alnifolia protects dopaminergic neurodegeneration in Caenorhabditis elegans

Context: The twigs of Sorbus alnifolia (Sieb. et Zucc.) K. Koch (Rosaceae) have been used to treat neuro- logical disorders as a traditional medicine in Korea. However, there are limited data describing the efficacy of S. alnifolia in Parkinson"s disease (PD).Objective: This study was conducted to identify the protective effects of the methanol extracts of S. alnifolia (MESA) on the dopaminergic (DA) neurodegeneration in Caenorhabditis elegans.Materials and methods: To test the neuroprotective action of MESA, viability assay was performed after 48 h exposure to 1-methyl-4-phenylpyridine (MMPþ) in PC12 cells and C. elegans (400 lM and 2 mM of MMPþ, respectively). Fluorescence intensity was quantified using transgenic mutants such as BZ555 (Pdat-1::GFP) and and UA57 (Pdat-1::GFP and Pdat-1::CAT-2) to determine MESA"s effects on DA neurode- generation in C. elegans. Aggregation of a-synuclein was observed using NL5901 strain (unc-54p::a- synuclein::YFP). MESA"s protective effects on the DA neuronal functions were examined by food-sensing assay. Lifespan assay was conducted to test the effects of MESA on the longevity.Results: MESA restored MPPþ-induced loss of viability in both PC12 cells and C. elegans (85.8% and 54.9%, respectively). In C. elegans, MESA provided protection against chemically and genetically-induced DA neurodegeneration, respectively. Moreover, food-sensing functions were increased 58.4% by MESA in the DA neuron degraded worms. MESA also prolonged the average lifespan by 25.6%. However, MESA failed to alter a-synuclein aggregation.Discussion and conclusions: These results revealed that MESA protects DA neurodegeneration and recov- ers diminished DA neuronal functions, thereby can be a valuable candidate for the treatment of PD

A Biodegradable Secondary Battery and its Biodegradation Mechanism for Eco-Friendly Energy-Storage Systems

The production of rechargeable batteries is rapidly expanding, and there are going to be new challenges in the near future about how the potential environmental impact caused by the disposal of the large volume of the used batteries can be minimized. Herein, a novel strategy is proposed to address these concerns by applying biodegradable device technology. An eco-friendly and biodegradable sodium-ion secondary battery (SIB) is developed through extensive material screening followed by the synthesis of biodegradable electrodes and their seamless assembly with an unconventional biodegradable separator, electrolyte, and package. Each battery component decomposes in nature into non-toxic compounds or elements via hydrolysis and/or fungal degradation, with all of the biodegradation products naturally abundant and eco-friendly. Detailed biodegradation mechanisms and toxicity influence of each component on living organisms are determined. In addition, this new SIB delivers performance comparable to that of conventional non-degradable SIBs. The strategy and findings suggest a novel eco-friendly biodegradable paradigm for large-scale rechargeable battery systems

Requirement of NV for inhibition of poly I∶C-induced IFN response in RTG-2 cells.

<p><i>A</i>, Effect of poly I∶C pre-treatment on the growth of rIHNVs. RTG-2 cells were pre-incubated with poly I∶C at 25 µg/ml for 24 h. The cells were then infected with rIHNV-32/87 or rIHNV-32/87-ΔNV-GFP at an MOI of 0.01 PFU/cell and samples of the supernatant medium were collected at 0, 24, and 48 h p.i. All data points represent the average of samples taken from duplicate infections. <i>B</i>, Effect of poly I∶C treatment after virus infection on the growth of rIHNVs. RTG-2 cells were infected with rIHNV-32/87 or rIHNV-32/87-ΔNV-GFP at an MOI of 0.01 PFU/cell. After incubation for 24 h, cells were washed three times and treated with 25 µg/ml of poly I∶C or serum-free media. At 0 and 24 h after poly I∶C treatment, samples of the supernatant medium were collected and titrated in duplicate. The virus titer in the supernatant medium collected at 0 h after poly I∶C treatment was defined as one. The results are presented as the means ± SD of three independent experiments (***P<0.001). ns, not significant. <i>C</i> and <i>D</i>, Analysis of IFN1 and Mx1 expressions in RTG-2 cells treated with poly I∶C after virus infection. RTG-2 cells were infected with rIHNV-32/87 or rIHNV-32/87-ΔNV-GFP at an MOI of 1 PFU/cell. After incubation for 24 h, cells were incubated with culture media containing 25 µg/ml of poly I∶C. At 24 h after poly I∶C treatment, total RNA was extracted from the cells and analyzed with real-time PCR for IFN1 (<i>C</i>) and Mx1 (<i>D</i>). The negative control was mock-infected RTG-2 cells without poly I∶C treatment. The positive control was mock-infected RTG-2 cells stimulated with 25 µg/ml poly I∶C. The levels of IFN1 and Mx1 are expressed as mRNA copy number normalized to 1000 copies of ARP mRNA. The results are presented as the means ± SD of three independent experiments (**P<0.01; ***P<0.001).</p