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>

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.

L-Deprenyl exerts cytotoxicity towards acute myeloid leukemia through inhibition of mitochondrial respiration

Ryu, I.,Ryu, M. J.,Han, J.,Kim, S. J.,Lee, M. J.,Ju, X.,Yoo, B. H.,Lee, Y. L.,Jang, Y.,Song, I.-C. Spandidos Publications 2018 Oncology reports Vol.40 No.6

Physics-based integrated modeling of the energy confinement time scaling laws for the H- and L-modes in the KSTAR-type tokamak model

Kim, J.Y.,Han, H.S.,Terzolo, L. International Atomic Energy Agency 2017 Nuclear fusion Vol.57 No.7

<P>In an effort to clarify the physics origin of the energy confinement time scaling law in H-mode plasmas, a new analysis method is first proposed where the stored energy is separated into two parts—one coming from the marginal stability with the pedestal boundary condition and the other related to the turbulent dynamics. The method is then applied for the analysis of the global scaling law, as initial examples, focusing on the four parameters of plasma current, input power, magnetic field and density in the KSTAR-type tokamak model. It is shown that the method can provide more quantitative and explicit information on how various physics elements, such as the linear stability, nonlinear turbulent dynamics and pedestal boundary, contribute to the global scaling factor. While this method is not directly applicable, the L-mode is also considered for comparison, trying to clarify how a difference in the scaling law can occur between the H- and L-modes.</P>

Characterization of the major dehydrogenase related to d-lactic acid synthesis in Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293

Li, L.,Eom, H.J.,Park, J.M.,Seo, E.,Ahn, J.E.,Kim, T.J.,Kim, J.H.,Han, N.S. IPC Science and Technology Press ; Elsevier Scienc 2012 Enzyme and microbial technology Vol.51 No.5

Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293 is a lactic acid bacterium that converts pyruvate mainly to d-(-)-lactic acid by using d-(-)-lactate dehydrogenase (ldhD). The aim of this study was to identify the gene responsible for d-lactic acid formation in this organism and to characterize the enzyme to facilitate the production of optically pure d-lactic acid. A genomic analysis of L. mesenteroides ATCC 8293 revealed that 7 genes encode lactate-related dehydrogenase. According to transcriptomic, proteomic, and phylogenetic analyses, LEUM_1756 was the major gene responsible for the production of d-lactic acid. The LEUM_1756 gene, of 996bp and encoding 332 amino acids (36.5kDa), was cloned and overexpressed in Escherichia coli BL21(DE3) Star from an inducible pET-21a(+) vector. The enzyme was purified by Ni-NTA column chromatography and showed a specific activity of 4450U/mg, significantly higher than those of other previously reported ldhDs. The gel permeation chromatography analysis showed that the purified enzyme exists as tetramers in solution and this was the first report among lactic acid bacteria. The pH and temperature optima were pH 8.0 and 30<SUP>o</SUP>C, respectively, for the pyruvate reduction reaction, and pH 11.0 and 20<SUP>o</SUP>C, respectively, for the lactate oxidation reaction. The K<SUB>m</SUB> kinetic parameters for pyruvate and lactate were 0.58mM and 260mM, respectively. In addition, the k<SUB>cat</SUB> values for pyruvate and lactate were 2900s<SUP>-1</SUP> and 2280s<SUP>-1</SUP>, respectively. The enzyme was not inhibited by Ca<SUP>2+</SUP>, Co<SUP>2+</SUP>, Cu<SUP>2+</SUP>, Mg<SUP>2+</SUP>, Mn<SUP>2+</SUP>, Na<SUP>+</SUP>, or urea, but was inhibited by 1mM Zn<SUP>2+</SUP> and 1mM SDS.

Effects of Expander Operating Conditions on Nutrient Digestibility in Finishing Pigs

S.L., Traylor,K.C., Behnke,J.D., Hancock,R.H., Hines,S.L., Johnston,B.J., Chae,In K., Han Asian Australasian Association of Animal Productio 1999 Animal Bioscience Vol.12 No.3

Five experiments were conducted using finishing pigs (PIC L326 sires $\times$ C15 dams) to examine the effects of expander operating conditions on nutrient digestibility in finishing pigs. The effects of different expanding conditions (0, 11.7, 24.4, $32.5kg/cm^2$) for corn-SBM based diets (Exp. 1), wheat meddlings diet (Exp. 2), sorghum-SBM based diets (Exp. 3) and wheat-SBM based diet (Exp. 4). Exp. 5 was conducted as a $2{\times}4$ factorial arrangement and factors examined were 2 soy products (raw soybean and SBM) and 4 expanding conditions (0, 14.1, 28.1, $42.2kg/cm^2$). In experiment 1, total production rates (p>0.10) were similar among treatments. The amount of fines decreased (cubic effect, p<0.001) as cone pressure was increased from 0 to $11.7kg/cm^2$, with smaller differences as cone pressure was further increased to $35.2kg/cm^2$. Nutrient digestibilities increased (p<0.02) as the feed was subjected to higher cone pressures. Digestibilities of DM, N, and GE were maximized at $24.4kg/cm^2$ cone pressure. The DE of the diet expanded at 24.4 and $35.2kg/cm^2$ increased by 172 and 109 kcal/kg, respectively, compared to the diet processed at $0kg/cm^2$ cone pressure. In experiment 2, total production and screened pellets production rates were similar among the processing treatments (p>0.21). The amount of fines decreased (quadratic effect, p<0.03) by 9 kg/h as cone pressure was increased from 0 to $11.7kg/cm^2$. Digestibilities of DM (p<0.02), N (p<0.001), and GE (p<0.002) were increased as cone pressure was increased from 0 to $35.2kg/cm^2$. DM, N, and GE digestibility in the pigs fed the midds-based diet increased by 8, 13, and 10%, respectively, at the highest processing cone pressure compared to the diets without any cone pressure. In experiment 3, the conditioned mash moistures for the treatments were numerically similar around 15% moisture. As the expander cone pressure was increased from 0 to $11.7kg/cm^2$, energy consumption for the pellet mill decreased (quadratic effect, p<0.004) from 14.1 to 12.0 kWh/t. Dry matter and gross energy digestibility increased (cubic effects, p<0.006) as cone pressure was increased from 0 to $35.2kg/cm^2$ with the largest improvement occurring as cone pressure was increased from 0 to $11.7kg/cm^2$. Nitrogen digestibility increased (cubic effect, p<0.001) from 78.3 to 81.0% as the feed was subjected to the higher cone pressures, with N digestibility being maximized at $24.4kg/cm^2$ cone pressure. The DE of the diet increased (cubic effect, p<0.001) by 225 kcal/kg as cone pressure was increased from 0 to $11.7kg/cm^2$. In experiment 4, pellet moisture decreased and moisture loss increased as cone pressure was increased from 0 to $35.2kg/cm^2$. Also, starch gelatinization of the wheat-based diets increased from 16.8 to 49.1% as the diets were processed at 0 and $35.2kg/cm^2$ cone pressure. Nutrient digestibilities were not affected (p>0.18) by any increase in cone pressure. In experiment 5, pellet moisture decreased as cone pressure was increased 0 to $35.2kg/cm^2$. The amount of moisture loss for the diets expanded at $42.2kg/cm^2$ was 3.0 and 3.8% for the SBM and raw soybean (RB) diets, respectively. Starch gelatinization for the SBM diets were 19% greater than the RB diets. The RB diets had lower DM, N and GE digestibilities as compared to the SBM diets. The DE of the RB diets were lower (p<0.02) than the SBM diets. DM (p<0.06), N (p<0.02), and GE (p<0.001) digestibilities of the dietary treatments increased as cone pressure was increased 0 to $42.2kg/cm^2$.

A Pan-Cancer Analysis of Enhancer Expression in Nearly 9000 Patient Samples

Chen, Han,Li, Chunyan,Peng, Xinxin,Zhou, Zhicheng,Weinstein, John N.,Caesar-Johnson, Samantha J.,Demchok, John A.,Felau, Ina,Kasapi, Melpomeni,Ferguson, Martin L.,Hutter, Carolyn M.,Sofia, Heidi J.,Ta Elsevier 2018 Cell Vol.173 No.2

<P><B>Summary</B></P> <P>The role of enhancers, a key class of non-coding regulatory DNA elements, in cancer development has increasingly been appreciated. Here, we present the detection and characterization of a large number of expressed enhancers in a genome-wide analysis of 8928 tumor samples across 33 cancer types using TCGA RNA-seq data. Compared with matched normal tissues, global enhancer activation was observed in most cancers. Across cancer types, global enhancer activity was positively associated with aneuploidy, but not mutation load, suggesting a hypothesis centered on “chromatin-state” to explain their interplay. Integrating eQTL, mRNA co-expression, and Hi-C data analysis, we developed a computational method to infer causal enhancer-gene interactions, revealing enhancers of clinically actionable genes. Having identified an enhancer ∼140 kb downstream of PD-L1, a major immunotherapy target, we validated it experimentally. This study provides a systematic view of enhancer activity in diverse tumor contexts and suggests the clinical implications of enhancers.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Systematic analysis of enhancer expression across ∼9,000 samples of 33 cancer types </LI> <LI> Global enhancer activation positively correlates with aneuploidy but not mutations </LI> <LI> A computational method that infers causal enhancer-target-gene relationships </LI> <LI> Enhancers as key regulators of therapeutic targets, including PD-L1 </LI> </UL> </P> <P><B>Graphical Abstract</B></P> <P>[DISPLAY OMISSION]</P>

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>

68Ga-PSMA PET/CT Imaging Predicting Intraprostatic Tumor Extent, Extracapsular Extension and Seminal Vesicle Invasion Prior to Radical Prostatectomy in Patients with Prostate Cancer

Christoph-Alexander J. von Klot,Axel S. Merseburger,Alena Böker,Sebastian Schmuck,Tobias L. Ross,Frank M. Bengel,Markus A. Kuczyk,Christoph Henkenberens,Hans Christiansen,Hans-Jürgen Wester,Wiebke Sol 대한핵의학회 2017 핵의학 분자영상 Vol.51 No.4

Purpose 68Ga-labeled prostate-specific membrane antigen (PSMA) ligand positron emission tomography/computed tomography (PET/CT) has shown promising results in patients with biochemical recurrence after primary therapy for prostate cancer. In this study, we evaluated the usefulness of PSMA I&T (imaging and therapy) PET/CT prior to radical prostatectomy. Methods The study population consisted of 21 patients with prostate cancer who underwent 68Ga-PSMA I&T PET/CT before either open or laparoscopic radical prostatectomy. Intraprostatic tumor extent, extracapsular extension (ECE) and seminal vesicle invasion (SVI) were assessed on the PET/CT scans. Tracer uptake was quantified in terms of standardized uptake values (SUVs). Imaging findings were correlated with final whole-gland histopathology. Results Of the 21 patients, two had T stage 2b disease, nine stage 2c, six stage 3a and four stage 3b. The median Gleason score was 7. The SUVmean of the primary tumors was 9.5 ± 8.8. SUVmean was higher in tumors with ECE than in organconfined tumors (13.8 ± 11.0 vs. 5.6 ± 3.2, p = 0.029). Peak tracer uptake was significantly positively correlated with Gleason score (rs = 0.49, p = 0.025). Sensitivity, specificity, positive predictive value and negative predictive value were, respectively, 94.7%, 75.0%, 97.3% and 60.0% for tumor infiltration of an individual prostate lobe, 75.0%, 100.0%, 100.0% and 97.4% for SVI, and 90.0%, 90.9%, 90.0% and 90.9% for ECE, using an angulated contour of the prostate as the criterion. Tumor volume derived from 68Ga-PSMA I&T PET/CT was significantly correlated with preoperative prostate-specific antigen value (rp = 0.75, p < 0.001) and tumor volume on histopathology (rp = 0.45, p = 0.039). Conclusions 68Ga-PSMA I&T PET/CT prior to radical prostatectomy can contribute to presurgical local staging of prostate cancer. In this pilot study, 68Ga-PSMA I&T PET/CT showed promising results for prediction of lobe infiltration, ECE and SVI.

여대생의 인유두종 바이러스 및 인유두종 바이러스 백신 관련 지식과 예방접종 실태와의 상관관계

Xu, Meiling,김현지,이희수,이윤지,한동연,박소영,엄효윤,최정민,홍서진,홍예림,이경은,이혜림 이화여자대학교 간호과학대학 2017 이화간호학회지 Vol.- No.51

Purpose: Human Papillomavirus (HPV) vaccination is readily available, but immunization rate is minimal. Lack of knowledge concerning the HPV infection and vaccine is expected as the primary reason. The correlation between knowledge levels and vaccination will provide applicable guidelines for successful interventions. Method: 309 female university students have completed surveys September 2016, reporting of HPV infection knowledge, HPV vaccine knowledge, source of knowledge, and attitudes concerning vaccination. Result: HPV infection and vaccine knowledge levels were both suboptimal(9.57±1.84, 7.59±1.58), and the Internet(16.2%) was recognized as the most generally used means of obtaining information on the HPV vaccine. However, vaccination rates were reported to be low(28.2%), with lack of information(46.6%) hindering immunization. Correlations were found between HPV infection knowledge and cervical cancer insight, but the correlations between knowledge levels and vaccination rates were insignificant. Conclusion: Prior information on cervical cancer had an effect on the knowledge levels of HPV infection, but other factors were insignificant. Lack of information on the HPV vaccine was speculated, thus programs should focus on accessibility and effectiveness.