Reduction of d-lactate content in sauerkraut using starter cultures of recombinant Leuconostoc mesenteroides expressing the ldhL gene

Jin, Q.,Li, L.,Moon, J.S.,Cho, S.K.,Kim, Y.J.,Lee, S.J.,Han, N.S. Society for Bioscience and Bioengineering, Japan ; 2016 Journal of bioscience and bioengineering Vol.121 No.5

<P>The n-form of lactate, which causes metabolic stress upon excessive dietary intake, is mainly produced by Leuconostoc sp., the predominant species in sauerkraut. To shift the metabolic flux of D-lactate from pyruvate to L-lactate, we expressed the L-lactate dehydrogenase (ldhL) gene in Leuconostoc mesenteroides ATCC 8293. The IdhL gene from Lactobacillus plantarum was introduced into L. mesenteroides using the shuttle vectors pLeuCM and pLeuCM42. To elevate the expression level of IdhL in L. mesenteroides, the nucleotides for pyruvate kinase promoter were fused to IdhL and cloned into above vectors to construct pLC18pkL and pLC42pkL. As results, introduction of pLC42pkL in L. mesenteroides significantly improved both L-LDH activity and L-lactate productivity during fermentation, decreasing the D-/L-lactate ratio. When used as a starter culture for sauerkraut fermentation, recombinant L. mesenteroides harboring pLC42pkL increased L-lactate concentration and decreased D-lactate concentration compared to the wild type strain. We newly developed a recombinant L. mesenteroides which has high L-lactate dehydrogenase activity and applied this strain to minimize the harmful effect of D-lactate during the sauerkraut fermentation. To the best of our knowledge, we demonstrate for the first time the effective use of recombinant Leuconostoc sp. for quality improvement of fermented foods. (C) 2015, The Society for Biotechnology, Japan. All rights reserved.</P>

High Sustained Virologic Response with Daclatasvir plus Asunaprevir in HCV GT-1b Chinese, Korean and Taiwanese without Baseline NS5A Polymorphisms

( F. Mcphee ),( L. Wei ),( Q. Xie ),( Y. Suzuki ),( J. Toyota ),( Y. Karino ),( K. Chayama ),( Y. Kawakami ),( M. L. Yu ),( S. H. Ahn ),( N. Zhou ),( H. Kumada ) 대한간학회 2016 춘·추계 학술대회 (KASL) Vol.2016 No.1

Aims: Daclatasvir (DCV) plus asunaprevir (ASV) has demonstrated highsustained virologic response (SVR) in HCV genotype (GT-)1b infection.NS5A-Y93H and NS5A-L31 resistance-associated polymorphisms(RAPs) to DCV are known to impact DCV+ASV response in GT-1b-infectedJapanese. The effect of RAPs on SVR at posttreatment week12 (SVR12) to DCV+ASV was explored in mainland Chinese, Korean,and Taiwanese.Methods: Pooled data from 2 studies of DCV (60 mg daily) + ASV(100 mg capsule, twice-daily) for 24 weeks in GT-1b-infected interferon/ribavirin-naive and -experienced patients from mainland China,Korea, and Taiwan. Similar Japanese data (4 studies; n=445) werepooled for comparison. SVR12 with versus without baseline Y93Hand/or L31 RAPs was compared by age (<65 vs ≥65 years), cirrhosisstatus, and baseline HCV-RNA.Results: SVR12 and baseline NS5A sequences were available for 282patients (126 mainland Chinese [45%〕, 80 Koreans [28%〕, 76Taiwanese [27%〕). NS5A-Y93H and/or -L31 RAPs were observed pretreatmentin 8% mainland Chinese, 14% Korean, and 18%Taiwanese patients, compared with 19% in Japanese. SVR12 in allnon-Japanese patients is shown (Figure); rates were broadly similarbetween countries and with Japanese data (Japanese: 96% overallwithout RAPs, 41% with RAPs). Responses were lower among patientswith baseline RAPs. By contrast, SVR12 in patients without RAPs washigh (92-100%), irrespective of cirrhosis, age, or baseline HCV-RNA.Conclusions: At least 95% of HCV GT-1b-infected patients from mainlandChina, Korea or Taiwan without baseline NS5A-Y93H or -L31polymorphisms who had HCV-RNA ≤7 log10 IU/mL achieved SVR12on DCV+ASV, regardless of cirrhosis status and age.

Effects of the novel angiotensin II receptor type I antagonist, fimasartan on myocardial ischemia/reperfusion injury

Han, J.,Park, S.J.,Thu, V.T.,Lee, S.R.,Long, L.T.,Kim, H.K.,Kim, N.,Park, S.W.,Jeon, E.S.,Kim, E.J.,Yoon, C.H.,Cho, G.Y.,Choi, D.J. Elsevier/North-Holland Biomedical Press 2013 INTERNATIONAL JOURNAL OF CARDIOLOGY Vol.168 No.3

Background: The aim of this study was to investigate the cardioprotective effect of fimasartan, a newly developed angiotensin II receptor type I blocker (ARB), against myocardial ischemia/reperfusion (I/R) injury and to identify the mechanism by which it reduces mitochondrial damage. Methods: Fimasartan was administered intravenously to Sprague-Dawley rats (3mg/kg), cardiomyocytes (50μM), and H9c2 cells (50μM) before ischemia or hypoxia. Myocardial infarction (MI), echocardiograms, DNA fragmentation, terminal deoxynucleotidyl transferase-mediated dUTP in situ nick-end labeling, immunoblotting, oxygen consumption, confocal microscopic appearance, and L-type Ca<SUP>2+</SUP> current (I<SUB>Ca,L</SUB>) were then assessed. Results: Fimasartan pretreatment remarkably reduced the rate of MI and improved cardiac performance well after I/R (n=9/group). Fimasartan also reduced apoptotic cell death both in vivo and in hypoxia/reoxygenation (H/R)-treated H9c2 cells (n=5~8/group). H/R-induced mitochondrial O<SUB>2</SUB><SUP>-</SUP> production and collapse of membrane potential were markedly attenuated in fimasartan-treated cardiomyocytes (n=4~6/group). Additionally, mitochondrial Ca<SUP>2+</SUP> overload during reoxygenation was suppressed by fimasartan (n=4~6/group), and this was found to be possibly related to the inhibition of I<SUB>Ca,L</SUB> and mitochondrial Ca<SUP>2+</SUP> uniporter. Furthermore, fimasartan pretreatment increased phosphorylations of Akt and glycogen synthase kinase-3β (n=5~7/group), decreased pro-apoptotic p53 levels, and increased anti-apoptotic Bcl-2 levels (n=4) during reperfusion. Conclusions: Fimasartan preconditioning has the potential to modulate Bcl-2 and suppress I/R-induced Ca<SUP>2+</SUP> overload by inhibiting I<SUB>Ca,L</SUB> and MCU. These beneficial effects could prevent the mitochondrial dysfunction and apoptosis accompanied by I/R.

서울시 지하철 구내의 공기중 분진 농도에 관한 연구

백남원(N.W. Paik),박두용(D.Y. Park),장익선(I.S. Chang),신용철(Y.C. Shin),이정인(J. l. Lee) 한국환경보건학회 1988 한국환경보건학회지 Vol.14 No.2

Airborne dust and asbestos fiber concentrations were determined in subway stations located in Seoul area. Two stations, such as Eulchiro 4.Ka Station of Une #2, constructed during a period of 1980-1984 and Hyehwa Station of Une #4, opened in 1985, were selected. The results of the study are as follows. 1. Daily tirne.weighted average (1W A) concentrations of airborne dusts from 07:00 to 20:00 hours in Line #2 and μne #4 were 0.43:t0.08 mg/m3 and 0.37:t0.12 mg/m3 , respectively. Thus, the dust levels in Líne #2 were significant1y higher than the levels in Line #4( p < 0.05). 2. Dust levels in the morning (07:00-11:00 hours), noon (1 1:00-16:00 hours) and in the evening (1 6:00-20:00 hours) in Líne #2 were 0.47:t0.17 mgjm3 , 0.37:t0.08 mg/m3 , and 0.46土0.07 mg/m3 respectively. Thus, dust levels in the morning and evening (i.e., during rush hours) were significantly higher than levels in the noon ( p < 0.02). However, there was no such difference in dust levels by time in Líne #4. 3. Airborne total dust concentrations were well below the occupational health standard of 10 mg/m3 , however, the levels were exceeding the ambient air standard recommended by the Korean Environment Administration. 4. All of airborne asbestos fiber concentrations were equal to or below 0.005 fibers/cc. The levels are within both occupational health standards and U .S. EPA criterìa. 5. A consideration should be given to the improvements of c1eaning methods (such as use of vacuum c1eaning instead of brushing) and the existing ventilation (such as more aìr change and filtration of supply air) for a reduction of dust levels. 6. It is recommended that routine monitoring of aìrborne dusts and asbestos fibers be conducted because more dusts and asbestos fibers can be produced in the aìr due to the deterioration of facilities by age and water damage in future.

Nitrogen Biofixing Bacteria Compensate for the Yield Loss Caused by ViralSatellite RNA Associated with Cucumber Mosaic Virus in Tomato

N. H. Dashti,M. S. Montasser,N. Y. Ali,R. G. Bhardwaj,D. L. Smith 한국식물병리학회 2007 Plant Pathology Journal Vol.23 No.2

To overcome the problem of the yield reduction due to the viral satellite mediated protection, a culture mix of three nitrogen-fixing bacteria species of the genus Azospirillum (A. brasilienses N040, A. brasilienses SP7, and A. lipoferum MRB16), and one strain of cyanobacteria (Anabena oryzae Fritsch) were utilized as biofertilizer mixture in both greenhouse and field experiments. When protected plants were treated with biofertilizer mixtures, the fruit yield of biofertilized plants increased by 48% and 40% in a greenhouse and field experiment, respectively, compared to untreated plants inoculated with the protective viral strain alone. Polyacrylamide gel electrophoresis (PAGE) analysis of total nucleic acid (TNA) extracts revealed that biofertilization did not affect the accumulation of the viral satellite RNA (CARNA 5) that is required for plant protection against other destructive viral strains of CMV. The yield increment was a good compensation for the yield loss caused by the use of the protective viral strain associated with CARNA 5.

Synthesis and evaluation of a novel chiral derivatization reagent for resolution of carboxylic acid enantiomers by RP-HPLC

Li, L.P.,Jin, M.N.,Shi, Q.,Xu, C.Y.,Jiang, Y.Z.,Lee, Y.I.,Min, J.Z. Academic Press 2017 Microchemical Journal Vol. No.

<P>A novel derivatization reagent, N-[1-Oxo-5-(triphenylphosphonium)pentyl]-(S)-3-aminopyrrolidine (OTPA), with triphenylphosphine (TPP) as a basic structure carrying a permanent positive charge was developed for the enantiomeric separation of chiral carboxylic acids by high-performance liquid chromatography (HPLC). OTPA reacted with the carboxylic acids at 40 degrees C within 90 min in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and 1-hydroxybenzotriazole (HOBt). The degree of epimerization (racemization) during the derivatization reaction was negligible. The separability of the diastereomers was evaluated in terms of their resolution value (Rs). The Rs values of the derivatives of non-steroidal anti-inflammatory drugs (NSAIDs), which were selected as the representative carboxylic acids, were completely separated by reversed-phase chromatography using an ODS (4.6 mm x 150 mm I.D., 5.0 mu m) column. The resolution Rs values were 1.54-2.23 for the evaluated carboxylic acids and the OTPA-derivatization was also effective for the enantiomeric separation of chiral carboxylic acids. The calibration curves (r(2) > 0.9971) were linear over the concentration range of 0.0125-1.25 mM for each enantiomer of ketoprofen (KET), and naproxen (NAP), 0.05-1.0 mM for each enantiomer of ibuprofen (IBU), 2-phenylpropionic acid (PPA), and loxoprofen (LOX), and 0.05-1.25 mM for each enantiomer of PBA. The limit of detection (S/N = 3) for each of the enantiomers of the NSAIDs and chiral carboxylic acid enantiomers was 1.4-7.6 mu mol/L. The inter-day and intra-day assay precisions were all <6.77% and the mean recoveries (%) of the NSAIDs and chiral carboxylic acids from the spiked human plasma were 95.27-101.12%. The derivatization followed by HPLC-UV enabled the separation and detection of NAP in human plasma with simple pretreatment. (C) 2017 Elsevier B.V. All rights reserved.</P>

Nitrogen Biofixing Bacteria Compensate for the Yield Loss Caused by Viral Satellite RNA Associated with Cucumber Mosaic Virus in Tomato

Dashti, N.H.,Montasser, M.S.,Ali, N.Y.,Bhardwaj, R.G.,Smith, D.L. The Korean Society of Plant Pathology 2007 Plant Pathology Journal Vol.23 No.2

To overcome the problem of the yield reduction due to the viral satellite mediated protection, a culture mix of three nitrogen-fixing bacteria species of the genus Azospirillum (A. brasilienses N040, A. brasilienses SP7, and A. lipoferum MRB16), and one strain of cyanobacteria (Anabena oryzae Fritsch) were utilized as biofertilizer mixture in both greenhouse and field experiments. When protected plants were treated with biofertilizer mixtures, the fruit yield of biofertilized plants increased by 48% and 40% in a greenhouse and field experiment, respectively, compared to untreated plants inoculated with the protective viral strain alone. Polyacrylamide gel electrophoresis (PAGE) analysis of total nucleic acid (TNA) extracts revealed that biofertilization did not affect the accumulation of the viral satellite RNA (CARNA 5) that is required for plant protection against other destructive viral strains of CMV. The yield increment was a good compensation for the yield loss caused by the use of the protective viral strain associated with CARNA 5.

Elimination of the cryptic plasmid in Leuconostoc citreum by CRISPR/Cas9 system

Jang, Y.J.,Seo, S.O.,Kim, S.A.,Li, L.,Kim, T.J.,Kim, S.C.,Jin, Y.S.,Han, N.S. Elsevier Science Publishers 2017 Journal of biotechnology Vol.251 No.-

<P>Leuconostoc spp. are important lactic acid bacteria for the fermentation of foods. In particular, L. citreum strains isolated from various foods have been used as host strains for genetic and metabolic engineering studies. In order to develop a food-grade genetic engineering system, L. citreum CB2567 was isolated from Kimchi. However, the isolated bacterium contained a cryptic plasmid which was difficult to eliminate. As the existence of the plasmid might hinder strain engineering, we eliminated the plasmid using an RNA-guided DNA endonuclease CRISPR/Cas9 system. We demonstrated that a plasmid-free L. citreum CB2567 host strain could be efficiently constructed through a two-step procedure: 1) transformation of the 'killer' plasmid expressing Cas9 endonuclease and a guide RNA (gRNA) targeting for a specific sequence in the cryptic plasmid, and 2) serial subculture without antibiotics for curing the killer plasmid. When the crude extract of L. citreum expressing Cas9 and the guide RNA was incubated with a PCR fragment containing the specific sequence recognized by the guide RNA, the PCR fragment was cleaved. Also, the cryptic plasmid pCB42 was successfully eliminated from the host strain after transforming the plasmid harboring Cas9 and the guide RNA. The Cas9 and gRNA expression plasmid used in this study can be applied for genome engineering purposes by additionally introducing an editing DNA template to repair the double strand DNA breakage caused by Cas9 in the genome of L. citreum. This study demonstrates the feasibility of developing CRISPR/Cas9-based genetic engineering tools to develop a safe host strain and construct food-grade lactic acid bacteria without residual antibiotic markers.</P>

Synthesis and characterization of Al-modified SBA-15 for Fischer–Tropsch synthesis (FTS) reaction

Kim, N. Y.,Jung, J. S.,Lee, J. S.,Yang, E. H.,Hong, G. H.,Lim, S. S.,Noh, Y. S.,Hodala, J. L.,Lee, K. Y.,Moon, D. J. Springer Science + Business Media 2016 Research on chemical intermediates Vol.42 No.1

<P>In this work, we synthesized a Co-based catalyst supported on modified SBA-15 to obtain the uniform particle size of active metal and selective products with certain carbon number distribution. For the comparison, modified SBA-15 was prepared with the different molar ratios of Si/Al = 5, 7 and 10 and compared with the SBA-15 support. Catalysts were prepared by the impregnation method and prepared catalysts were characterized by various analytical techniques such as N-2 physisorption, XRD, SAXS, TPR, NH3-TPD, FT-IR, and TEM. It was found that, by adding the aluminum to SBA-15, the catalyst acid site was increased and it produced a relatively middle range of hydrocarbons. It was also found that the interaction between cobalt metal and the support was increased, affecting the catalyst activity for Fischer-Tropsch synthesis.</P>

Comparison of Natural Resistance-associated Macrophage Protein (NRAMP)1 Expression between Cows with High and Low Milk Somatic Cells Counts

Joo, Y.S.,Moon, J.S.,Fox, L.K.,Suh, G.H.,Kwon, N.H.,Kim, S.H.,Park, Y.H. Asian Australasian Association of Animal Productio 2003 Animal Bioscience Vol.16 No.12

Studies using natural resistance-associated macrophage protein (NRAMP) identification indicated that cattle could be selected for immunity. Several studies performed on intracellular organisms such as Mycobacterium, Salmonella, Brucella and Leishmania in human and mouse revealed that resistance against these bacteria was dependent on high activity of NRAMP1 in macrophages. However, hardly any researches have been done on Staphylococcus aureus in bovine mastitis, which is an intracellular organism and the main cause of bovine mastitis. The objectives of this study were to establish reverse transcriptase polymerase chain reaction (RT-PCR) methods, through which NRAMP1 mRNA expression could be compared and analyzed between mastitis-resistant and -susceptible cows. NRAMP1 gene and its expression were investigated using 20 cows (Holstein Friesian) in Korea. Cows were evenly split into two groups, with and without histories of clinical mastitis. Equivalent numbers of cows were randomly selected from each group. Monocytes were isolated from the bovine peripheral blood of each selected cows and activated with lipopolysaccharide (LPS). mRNA was separated from the monocytes and cDNA of NRAMP1 was synthesized and amplified using RT-PCR with amplification of $\beta$-actin as a control. The difference in NRAMP1 expressions of mastitis-resistant (n=10) and -susceptible (n=10) Holstein cows was analyzed. Results demonstrate that resistant cows produced more NRAMP1 mRNA than the susceptible ones, and ratios of NRAMP1:$\beta$-actin expression were higher in resistant cows with or without LPS activation. Therefore, this study could be applied to select bovine mastitis resistant cows before infection based on the expression of NRAMP1.