• Title/Summary/Keyword: LU7

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Identification of Plasmid-Free Chlamydia muridarum Organisms Using a Pgp3 Detection-Based Immunofluorescence Assay

  • Chen, Chaoqun;Zhong, Guangming;Ren, Lin;Lu, Chunxue;Li, Zhongyu;Wu, Yimou
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1621-1628
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    • 2015
  • Chlamydia possesses a conserved 7.5 kb plasmid that is known to play an important role in chlamydial pathogenesis, since some chlamydial organisms lacking the plasmid are attenuated. The chlamydial transformation system developed recently required the use of plasmid-free organisms. Thus, the generation and identification of plasmid-free organisms represent a key step in understanding chlamydial pathogenic mechanisms. A tricolor immunofluorescence assay for simultaneously detecting the plasmid-encoded Pgp3 and whole organisms plus DNA staining was used to screen C. muridarum organisms selected with novobiocin. PCR was used to detect the plasmid genes. Next-generation sequencing was then used to sequence the genomes of plasmid-free C. muridarum candidates and the parental C. muridarum Nigg strain. We generated five independent clones of plasmid-free C. muridarum organisms by using a combination of novobiocin treatment and screening plaque-purified clones with anti-Pgp3 antibody. The clones were confirmed to lack plasmid genes by PCR analysis. No GlgA protein or glycogen accumulation was detected in cells infected with the plasmid-free clones. More importantly, whole-genome sequencing characterization of the plasmid-free C. muridarum organism and the parental C. muridarum Nigg strain revealed no additional mutations other than loss of the plasmid in the plasmid-free C. muridarum organism. Thus, the Pgp3-based immunofluorescence assay has allowed us to identify authentic plasmid-free organisms that are useful for further investigating chlamydial pathogenic mechanisms.

Overexpressions of xylA and xylB in Klebsiella pneumoniae Lead to Enhanced 1,3-Propanediol Production by Cofermentation of Glycerol and Xylose

  • Lu, Xinyao;Fu, Xiaomeng;Zong, Hong;Zhuge, Bin
    • Journal of Microbiology and Biotechnology
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    • v.26 no.7
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    • pp.1252-1258
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    • 2016
  • 1,3-Propanediol (1,3-PD) is a valuable platform compound. Many studies have shown that the supplement of NADH plays a key role in the bioproduction of 1,3-PD from Klebsiella pneumoniae. In this study, the xylA and xylB genes from Escherichia coli were overexpressed individually or simultaneously in K. pneumoniae to improve the production of 1,3-PD by cofermentation of glycerol and xylose. Compared with the parent strain, the xylose consumption was significantly increased by the introduction of these two genes. The 1,3-PD titers were raised from 17.9 g/l to 23.5, 23.9, and 24.4 g/l, respectively, by the overexpression of xylA and xylB as well as their coexpression. The glycerol conversion rate (mol/mol) was enhanced from 54.1% to 73.8%. The concentration of 2,3-butanediol was increased by 50% at the middle stage but drastically decreased after that. The NADH and NADH/NAD+ ratio were improved. This report suggests that overexpression of xylA or xylB is an effective strategy to improve the xylose assimilation rate to provide abundant reducing power for the biosynthesis of 1,3-PD in K. pneumoniae.

Enhanced 2,3-Butanediol Production in Recombinant Klebsiella pneumoniae via Overexpression of Synthesis-Related Genes

  • Kim, Borim;Lee, Soojin;Park, Joohong;Lu, Mingshou;Oh, Minkyu;Kim, Youngrok;Lee, Jinwon
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1258-1263
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    • 2012
  • 2,3-Butanediol (2,3-BD) is a major metabolite produced by Klebsiella pneumoniae KCTC2242, which is a important chemical with wide applications. Three genes important for 2,3-BD biosynthesis acetolactate decarboxylase (budA), acetolactate synthase (budB), and alcohol dehydrogenase (budC) were identified in K. pneumoniae genomic DNA. With the goal of enhancing 2,3-BD production, these genes were cloned into pUC18K expression vectors containing the lacZ promoter and the kanamycin resistance gene to generate plasmids pSB1-7. The plasmids were then introduced into K. pneumoniae using electroporation. All strains were incubated in flask experiments and 2,3-BD production was increased by 60% in recombinant bacteria harboring pSB04 (budA and budB genes), compared with the parental strain K. pneumoniae KCTC2242. The maximum 2,3-BD production level achieved through fed-batch fermentation with K. pneumoniae SGJSB04 was 101.53 g/l over 40 h with a productivity of 2.54 g/l.h. These results suggest that overexpression of 2,3-BD synthesis-related genes can enhance 2,3-BD production in K. pneumoniae by fermentation.

Monitoring Mount Sinabung in Indonesia Using Multi-Temporal InSAR

  • Lee, Chang-Wook;Lu, Zhong;Kim, Jin Woo
    • Korean Journal of Remote Sensing
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    • v.33 no.1
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    • pp.37-46
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    • 2017
  • Sinabung volcano in Indonesia was formed due to the subduction between the Eurasian and Indo-Australian plates along the Pacific Ring of Fire. After being dormant for about 400 years, Sinabung volcano erupted on the 29th of August, 2010 and most recently on the 1st of November, 2016. We measured the deformation of Sinabung volcano using Advanced Land Observing Satellite/Phased Array type L-band Synthetic Aperture Radar(ALOS/PALSAR) interferometric synthetic aperture radar(InSAR) images acquired from February 2007 to January 2011. Based on multi-temporal InSAR processing, we mapped the ground surface deformation before, during, and after the 2010 eruption with time-series InSAR technique. During the 3 years before the 2010 eruption, the volcano inflated at an average rate of ~1.7 cm/yr with a markedly higher rate of 6.6 cm/yr during the 6 months prior to the 2010 eruption. The inflation was constrained to the top of the volcano. From the 2010 eruption to January 2011,the volcano subsided by approximately 3 cm (~6 cm/yr). We interpreted that the inflation was due to magma accumulation in a shallow reservoir beneath Sinabung. The deflation was attributed to magma withdrawal from the shallow reservoir during the eruption as well as thermo-elastic compaction of erupted material. This result demonstrates once again the utility of InSAR for volcano monitoring.

A Novel Mannose-binding Tuber Lectin from Typhonium divaricatum (L.) Decne (family Araceae) with Antiviral Activity Against HSV-II and Anti-proliferative Effect on Human Cancer Cell Lines

  • Luo, Yongting;Xu, Xiaochao;Liu, Jiwei;Li, Jian;Sun, Yisheng;Liu, Zhen;Liu, Jinzhi;Damme, Els Van;Balzarini, Jan;Bao, Jinku
    • BMB Reports
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    • v.40 no.3
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    • pp.358-367
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    • 2007
  • A novel mannose-binding tuber lectin with in vitro antiproliferative activity towards human cancer cell lines and antiviral activity against HSV-II was isolated from fresh tubers of a traditional Chinese medicinal herb, Typhonium divaricatum (L.) Decne by a combined procedure involving extraction, ammonium sulfate precipitation, ion exchange chromatography on DEAE-SEPHAROSE, CM-SEPHAROSE and gel-filtration on sephacryl S-200. The apparent molecular mass of the purified Typhonium divaricatum lectin (TDL) was 48 kDa. TDL exhibits hemagglutinating activity toward rabbit erythrocytes at 0.95 $\mu$g/ml, and its activity could be strongly inhibited by mannan, ovomucoid, asialofetuin and thyroglobulin. TDL showed antiproliferative activity towards some well established human cancer cell lines, e.g. Pro-01 (56.7 $\pm$ 6.8), Bre-04 (41.5 $\pm$ 4.8), and Lu-04 (11.4 $\pm$ 0.3). The anti-HSV-II activity of TDL was elucidated by testing its HSV-II infection inhibitory activity in Vero cells with $TC_50$ and $EC_50$ of 5.176 mg/ml and 3.054 $\mu$g/ml respectively. The full-length cDNA sequence of TDL was 1145 bp and contained an 813-bp open reading frame (ORF) encoding a 271 amino acid precursor of 29-kDa. Homology analysis showed that TDL had high homology with many other mannose-binding lectins. Secondary and three-dimensional structures analyses showed that TDL is heterotetramer and similar with lectins from mannose-binding lectin superfamily, especially those from family Araceae.

Expression of Smoothened Protein in Colon Cancer and its Prognostic Value for Postoperative Liver Metastasis

  • Ding, Yin-Lu;Wang, Qi-San;Zhao, Wei-Min;Xiang, Lei
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4001-4005
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    • 2012
  • Backgrouds: The hedgehog (Hh) signaling pathway is composed of patched (PTCH) and smoothened (SMO), two transmembrane proteins, and downstream glioma-associated oncogene homolog (Gli) transcription factors. Hh signaling plays a pathological role in the occurrence and development of various cancers. Methods: To investigate the expression of SMO protein in colon cancer and its association with clinicopathological parameters and postoperative liver metastasis, immunohistochemistry was performed with paraffin-embedded specimens of 96 cases. Relationships between SMO protein expression and clinicopathological parameters, postoperative liver metastasis were analyzed. Results: IHC examination showed that SMO protein expression was significantly increased in colon cancer tissues compared to normal colon tissues (P = 0.042), positively related to lymph node metastases (P = 0.018) and higher T stages (P = 0.026). Postoperative live metastasis-free survival was significantly longer in the low SMO expression group than in those with high SMO expression ($48.7{\pm}8.02$ months vs $28.0{\pm}6.86$ months, P=0.036). Multivariate analysis showed SMO expression level to be an independent prognostic factor for postoperative live metastasis-free survival (95% confidence interval [CI] =1.46-2.82, P = 0.008). Conclusions: Our results suggest that in patients with colon cancer, the SMO expression level is an independent biomarker for postoperative liver metastasis, and SMO might play an important role in colon cancer progression.

Association Between EGF, TGF-β1 and TNF-α Gene Polymorphisms and Hepatocellular Carcinoma

  • Shi, Hai-Zhou;Ren, Peng;Lu, Qing-Jun;Niedrgethmnn, Marco;Wu, Guo-Yang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6217-6220
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    • 2012
  • Introduction: Up to present, EGF $61^*A$/G, TGF-${\beta}1$-$509^*T$/C and TNF-${\alpha}$-$308^*A$/G gene polymorphisms have been analysed in other cancer entities than hepatocellular carcinoma (HCC). We here investigated the frequency of these gene polymorphisms among HCC patients. Materials and Methods: A total of 73 HCC patients and 117 cancer-free healthy people were recruited at the Surgical Department of Zhongshan Hospital. Genomic DNA was isolated from peripheral blood and gene polymorphisms were analyzed by PCR-RFLP. Results: The distribution of EGF $61^*G$/G homozygotes among HCC patients was more frequent than that in the control group (24.7% vs 11.1%, OR=2.618, 95%CI=1.195-5.738). In parallel, the frequency of the "G" allele in the HCC patient group was also higher than that in the control group (45.9% vs 33.3%, OR= 1.696, 95%CI=1.110-2.592). No difference could be found for the TGF-${\beta}1$-509 and TNF-${\alpha}$-308 genotypes. Conclusion: EGF $61^*G$/G genotype and G allele are significantly increased among patients with HCC. TGF-${\beta}1$-$509^*T$/C and TNF-${\alpha}$-$308^*A$/G gene polymorphisms are not related to this cancer entity.

The Bibliogrphical Study on the Allergic Rhinitis (알레르기性 鼻炎에 對한 文獻的 考察)

  • Kim, Hyun-Ah;Jung, Ji-Chun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.7 no.1
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    • pp.53-84
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    • 1994
  • The study has been carried out to investigate of the Allergic Rhinitis by referring to 87 literatures. The results were as follows; 1. In oriental medical science, Allergic Rhinitis is belong to the category of the 'BiGu'(鼻구) 'GuChe'(구체). The 'Gu'(구) of the BiGu means watery rhinorrhea, the 'Che'(체) of the GuChe means sneezing. 2. The cause of a disease summarize the weak of Lung, Spleen and Kidney, and invasion into the nasal cavity of PoongHan etc a wrong air. Sometimes the pathologial change appear PoongHan changeHwa(風寒化火), HwaYeol is hidden in the inside(火熱內伏). The contributing factors are found a season(spring, winter), an abnormal weather(運氣 : 少陰 少陽 陽明 司天, 歲金不及), an emotional stress, an external wound of the harmful air, a food allergens and fatigue, a contact of substances, a sunlight etc. 3. Predominant symptoms are watery rhinorrhea, sneezing and nasal obstruction. Sometimes accompanic symptoms are nasal bleeding, mucopurulent rhinorrhea, olfactory disturbance, nasal polyp, rhinolalia clausa, respiratory disfunction etc. 4. The treatment-methodes is as follows, OnBoPaeJang GeoPoongSanHan(溫補肺臟 祛風散寒), GeonBilkGi(健脾益氣), BoSinNabGi (補腎納氣). The treatmentherbs is as follows, OnBoJiLuDan GaGam(溫補止流丹 加減), OkByeongPoongSan plus ChangIJaSan GaGam(玉屛風散合 蒼耳子散 加減), BoJungIkGiTang GaGam plus SoCheongLyongTang(補中益氣湯加減 配合 小靑龍湯), SinGiHwan GaGam(腎氣丸加減), GaeJiTang(桂枝湯) etc. 5. The external treatment is as follows, JeokBi(滴鼻), ChuiBi(吹鼻), SaekBi(塞鼻), stick and herbs-injection on the acupuncture-point, pressure ear acupuncture-point, herbs-pillow etc. 6. The acupuncture-moxa treatment is as follows, the methodes of cure apply TongJoGyeongGi(通調經氣), SanTongBiGui(宣通鼻竅) etc. Predominent acupuncture-points are YoungHyang(迎香), InDang(印堂), BiTong(鼻通), SangSeong(上星), HabGok(合谷) and so on. As mentioned above, from now on, it's need to the oriental medical scientific study of the Immunity and Allergy and to the external treatment's application for the ascent of the treatment-effect of the allergic disease.

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The Influence of Rapid Thermal Annealing Processed Metal-Semiconductor Contact on Plasmonic Waveguide Under Electrical Pumping

  • Lu, Yang;Zhang, Hui;Mei, Ting
    • Journal of the Optical Society of Korea
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    • v.20 no.1
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    • pp.130-134
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    • 2016
  • The influence of Au/Ni-based contact formed on a lightly-doped (7.3×1017cm−3, Zn-doped) InGaAsP layer for electrical compensation of surface plasmon polariton (SPP) propagation under various rapid thermal annealing (RTA) conditions has been studied. The active control of SPP propagation is realized by electrically pumping the InGaAsP multiple quantum wells (MQWs) beneath the metal planar waveguide. The metal planar film acts as the electric contact layer and SPP waveguide, simultaneously. The RTA process can lower the metal-semiconductor electric contact resistance. Nevertheless, it inevitably increases the contact interface morphological roughness, which is detrimental to SPP propagation. Based on this dilemma, in this work we focus on studying the influence of RTA conditions on electrical control of SPPs. The experimental results indicate that there is obvious degradation of electrical pumping compensation for SPP propagation loss in the devices annealed at 400℃ compared to those with no annealing treatment. With increasing annealing duration time, more significant degradation of the active performance is observed even under sufficient current injection. When the annealing temperature is set at 400℃ and the duration time approaches 60s, the SPP propagation is nearly no longer supported as the waveguide surface morphology is severely changed. It seems that eutectic mixture stemming from the RTA process significantly increases the metal film roughness and interferes with the SPP signal propagation.

Landscape Management Strategies Based on Landscape Assessment of Riparian Buffer Zone in the Han River: Focused on Gyeongan Stream (경관평가 기반 수변구역의 경관관리 전략 - 경안천 수변구역을 대상으로 -)

  • Park, Chang Sug;Bae, Min-Ki
    • Journal of Environmental Impact Assessment
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    • v.21 no.6
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    • pp.903-916
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    • 2012
  • We need to use aesthetic landscape assessment(ALA) as the means of riparian buffer zone(RBZ) management. This study verified the political validity of designation policy of RBZ and land purchase policy. The purpose of this study was to propose RBZ landscape management strategies through ALA of RBZ in Gyeongan Stream for the better attractive and healthy riparian landscapes. The natural type landscape units(LUs) covered 40.9% of the entire area and the cultivated land type LUs covered 20.58%. Landscape assessment consisted of landscape quality and landscape integration assessment. The criteria for assessing landscape quality(LQ) were naturalness, interest, uniqueness, and landscape function. LQ was ranked into five grades using a matrix. The landscape integration assessment consisted of an inner integration assessment in each LU and outer integration assessment among LUs. To review the propriety of designating the riparian area and the riparian ecological belt, differences in ecological appraisal and aesthetic valuation were reviewed through a t-test, Oneway ANOVA, and logistic analysis. The results of ALA, 29.15% of the entire area scored at grade 1 in LQ, while 31.95% scored at grade 5, indicating that grade 5 areas occupied a high share. Surveyed areas were divided into designated RBZ and undesignated RBZ. Results indicated that designated RBZ scored grade 1 in LQ took up 33.2% of the total, significantly higher than the 23.3% taken up by undesignated RBZ. When examined according to buffer distance, grade 1 areas within 50m took up 50.2% of the total area, lower than the 32.7% at buffer distances of 500m-1km. Results indicated a 1% statistically significant difference. Accordingly, analysis was undertaken for the expansion of designation of the riparian area and the selection of appropriate land for formation of a riparian ecological belt, and was designated at priority 1 and 2 for land purchase. This study can also contribute to the formation of a riverine eco-belt through discovery of design factors for upgrading the ecology, aesthetics, and landscape of the riparian area and application in determining land purchase priorities.