• Title/Summary/Keyword: Ala

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Homology Modeling and Active Sites of PolyMG-specific Alginate Lyase from Stenotrophomonas maltophilia KJ-2 (Stenotrophomonas maltophilia KJ-2 균주로부터 얻은 PolyMG-specific 알긴산분해효소의 상동성 모델링 및 활성자리 연구)

  • Kim, Hee Sook
    • Journal of Life Science
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    • v.24 no.2
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    • pp.128-136
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    • 2014
  • Alginates are linear acidic polysaccharides composed with (1-4)-linked ${\alpha}$-L-guluronic acid and ${\beta}$-Dmannuronic acid. Alginate can be degraded by diverse alginate lyases, which cleave the alginate using a ${\beta}$-elimination reaction and produce unsaturated uronate oligomers. A gene for a polyMG-specific alginate lyase possessing a novel structure was previously identified and cloned from Stenotrophomonas maltophilia KJ-2. Homology modeling of KJ-2 polyMG-specific alginate lyase showed it belongs to the PL6 family, whereas three Azotobacter vinelandii polyMG lyases belong to the PL7 family of polysaccharide lyases. From $^1H$-NMR spectra data, KJ-2 polyMG lyase preferably degraded the M-${\beta}$(1-4)-G glycosidic bond than the G-${\alpha}$(1-4)-M glycosidic bond. Seventeen mutants were made by site-directed mutagenesis, and alginate lyase activity was analyzed. Lys220Ala, Arg241Ala, Arg241Lys, and Arg265Ala lost alginate lyase activity completely. Arg155Ala, Gly303Glu, and Tyr304Phe also lost the activity by 60.7-80.1%. These results show that Arg155, Lys220, Arg241, Arg265, Gly303, and Tyr304 are important residues for catalytic activity and substrate binding.

Effect of glutamic acid and its ${\gamma}-derivatives$ on the production of ${\delta}-aminolevulinic{\;}acid$ by Rhodobacter sphaeroides (Rhodobacter sphaeroides에 의한 ${\delta}-aminolevulinic{\;}acid$생산에 있어서 glutamic acid 및 감마 유도체의 영향)

  • Choi, Kyung-Min;Lim, Wang-Jin;Hwang, Se-Young
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.184-190
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    • 1993
  • The effect of ${\delta}-aminolevulinic\;acid$ (ALA) biosynthetic precursors and related compounds on the ALA productivity from a strain of Rhodobacter sphaeroides has been examined in vivo and in vitro systems. The relative ratios of ALA productivities by $C_{4}$- pathway to that by $C_{5}$-pathway in vivo and in vitro systems were 0.78 and 1.37, respectively. Although the expression rates of $C_{4}-$ and $C_{5}-pathways$ in cell-free systems prepared after precursors supplemented cultivations were increased 1.35 and 1.52 folds, respectively, the rate increase of $C_{4}-pathway$ was accompanied by the rate decrease of the $C_{5}-pathways$, and vice versa, as that the rates of both $C_{4}-$ and $C_{5}-pathways$ were lowered to be 0.91, 0.83, respectively. The order of cellular uptake rates of ${\gamma}-glutamyl$ derivatives relative to that found with L-glutamic acid were shown to be D-glutamic acid, 0.55: D-glutamine, 0.5: L-glutamine, 0.4: ${\gamma}-L-glutamyl$ ethylester, 0.3: GSH and Glu-pNA, 0. L and D configurations of glutamine were indicated as better substrates in vivo for ALA yields than those of glutamic acid, respectively.

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Study of a Brain Tumor and Blood Vessel Detection System Using Multiple Fluorescence Imaging by a Surgical Microscope (수술현미경에서의 다중형광영상을 이용한 뇌종양과 혈관영상 검출 시스템 연구)

  • Lee, Hyun Min;Kim, Hong Rae;Yoon, Woong Bae;Kim, Young Jae;Kim, Kwang Gi;Kim, Seok Ki;Yoo, Heon;Lee, Seung Hoon;Shin, Min Sun;Kwon, Ki Chul
    • Korean Journal of Optics and Photonics
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    • v.26 no.1
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    • pp.23-29
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    • 2015
  • In this paper, we propose a microscope system for detecting both a tumor and blood vessels in brain tumor surgery as fluorescence images by using multiple light sources and a beam-splitter module. The proposed method displays fluorescent images of the tumor and blood vessels on the same display device and also provides accurate information about them to the operator. To acquire a fluorescence image, we utilized 5-ALA (5-aminolevulinic acid) for the tumor and ICG (Indocyanine green) for blood vessels, and we used a beam-splitter module combined with a microscope for simultaneous detection of both. The beam-splitter module showed the best performance at 600 nm for 5-ALA and above 800 nm for ICG. The beam-splitter is flexible to enable diverse objective setups and designed to mount a filter easily, so beam-splitter and filter can be changed as needed, and other fluorescent dyes besides 5-ALA and ICG are available. The fluorescent images of the tumor and the blood vessels can be displayed on the same monitor through the beam-splitter module with a CCD camera. For ICG, a CCD that can detect the near-infrared region is needed. This system provides the acquired fluorescent image to an operator in real time, matching it to the original image through a similarity transform.

Photodynamic Therapy induced Cell Death using ALA and 632nm Diode Laser in A549 Lung Cancer Cells (A549 폐암세포주에서 ALA와 632nm Diode Laser를 이용한 광역학치료 유도성 세포사)

  • Kim, Youn Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.2
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    • pp.178-186
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    • 2004
  • Background : Photodynamic therapy (PDT) is a new therapeutic method aimed at the selective destruction of cancer cells. The outcome is death of cancer cells through apoptosis or necrosis. The aim of this study was to investigate the characterization of PDT induced cell death in A549 lung cancer cells. Materials and methods : A549 cells were used as the lung cancer cell. 5 aminolevulinic acid (ALA) was used as the photosensitizer and a 632nm diode laser (Biolitec, Germany) as the light source. Cells were incubated with various concentrations of ALA. The 632nm diode laser was then administered for various laser irradiation times. The treated cells were incubated with 24, 48 and 72 hours. The cell viabilities were measured using the crystal violet assay and light microscopy. To observe the cell death mechanism after PDT, cells were observed under fluorescence microscopy after double staining with Hoechst 33342 and propium iodide after PDT. Results : In the crystal violet assay at 24 hours after PDT with a $3.2J/cm^2$ laser irradiation power, the cell viabilities were $89.56{\pm}4.11$, $87.67{\pm}5.48$, and $69.37{\pm}8.84$ with ALA concentrations of 10, 100, and $1mg/m{\ell}$, respectively. In crystal violet assay at 24 hours after PDT with $1mg/m{\ell}$ of ALA, the cell viabilities were $74{\pm}19.85$, $55{\pm}6.1$, and $49.06{\pm}16.64%$ with 1.6, 3.2 and $6.4J/cm^2$ laser irradiation powers, respectively. However, increasing the interval time after PDT did not change the cell viabilities. In the apoptosis assay, photodynamic therapy was inducing the apoptotic cell death. Conclusions : This study shows the apoptotic anticancer effect of photodynamic therapy in A549 lung cancer cells. However, further evaluations with other cancer cells and photosensitizers are necessary.

Manufacturing artificial lightweight aggregates using coal bottom ash and its application to the lightweight-concretes (석탄 바닥재를 이용한 인공경량골재의 제조 및 경량 콘크리트에 적용)

  • Kim, Kang-Duk;Kang, Seung-Gu
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.18 no.5
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    • pp.211-216
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    • 2008
  • The artificial lightweight aggregate (ALA) was manufactured in a rotary kiln at $1125^{\circ}C$ using green body formed by pelletizing the batch powder composing of coal bottom ash (CBA) produced from power plant, clay and dredged soil (DS). The TCLP (Toxicity characteristic leaching procedure) results showed that the dissolution concentration of heavy metal ions of ALA fabricated in this study was below the limitation defined by the enforcement regulations of wastes management law in Korea. The ALA containing 60$\sim$70 wt% CBA had a bulk density of 1.45$\sim$1.49 and a water absorption of 17.2$\sim$18.5 %. The impact values for oven-dry state and saturated-surface dry state of ALA were 27.4$\pm$1.3 and 23.4$\pm$2.6 % respectively. The 28-days compressive strength of concrete made with various ALA was $22.7\sim27.8 N/mm^2$. The slump of concrete with ALA containing CBA 60 and 70 wt% were 7.9 and 14.3 cm respectively. The unit weight of concrete made with any ALA fabricated in this study was satisfied with the standard specifications of lightweight concrete for the civil engineering and construction presented by Korea as below $1.84 ton/m^3$.

Industrial Utilization and Function of Omega Fatty Acid and Their Content Variation in Perilla (들깨 오메가 지방산의 기능과 함량변이 및 이용)

  • Ryu Su Noh;Lee Seung Tack;Lee Jung Il;Lee Jae Hak
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.spc1
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    • pp.110-122
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    • 1996
  • The n-3 family fatty acids containing ${\alpha}$-linolenic acid(18:3, ALA) have been known as physiological activation materials such as inhibitory effects on the incidence of hyper-tension, coronary heart disease and cancers as well as the control of senilc dementia. Although a lot of ALA(about $63\%$) are contained in perilla oil, it has not been commercialized yet because the purification technique of the ALA has not been well established. The procedure of purification of ALA from perilla oil was saponified with 1 N-KOH /ethanol and then saturated and low level unsaturated fatty acids were removed by low-temperature crystallization method. The concentrated unsaturated fatty acids (containing about $75\%$ ALA) went down through the silver nitrate-impregnated silica column chromatography for separation of high purity of ALA. The results obtained we Fraction B, C and D contained ALA more than $85.5\%$(recovery, >$88.9\%,\;95.4\%$(recovery, >$54.4\%$) and $99.9\%$(recovery, >$31.5\%$) in purity, respectively. Seed oil content of the tested varieties were ranged from 34.8 to $54.1\%$ with $45.3\%$ of varietal means. The major omega fatty acids contained in the oil were oleic acid(n-9) $15.2\%$, linoleic acid(n-6) $13.9\%$ and linolenic acid(n-3) $63.1\%$ in the mean value. Varietal variation of n-9, 6 and 3 fatty acids ranged of $9.5\~21.4\%,\;9.1\~20.4\%$ and $50.6\~70.5\%$ respectively. Unsaturated fatty acid were averaged $92.2\%$ of seed oil in fatty acid composition. The ratios of n-6 to n-3 ranged of $0.13\~0.34\%$($0.22\%$ in mean value). The highest n-3 fatty acid variety was Yecheonjong being $70.5\%$. The lowest variety in ratios of n-6 to n-3 was Goseongjong being $0.13\%$. Oil content showed positive correlation with stearic acid and linolenic acid, while the negative correlation with oil content and linoleic acid. On the other hand, A significant negative correlation were showed between linolnic acid and the ratios n-6/n-3 fatty acid, saturated fatty acid. Saturated fatty acid was highly correlated with unsaturated fatty acid negatively being $r= -0.723^{**}$.

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A Strategy Using Photodynamic Therapy and Clofibric Acid to Treat Peritoneal Dissemination of Ovarian Cancer

  • Yokoyama, Yoshihito;Shigeto, Tatsuhiko;Miura, Rie;Kobayashi, Asami;Mizunuma, Makito;Yamauchi, Aisa;Futagami, Masayuki;Mizunuma, Hideki
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.2
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    • pp.775-779
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    • 2016
  • Background: The current study examined the effectiveness of concurrent therapy using photodynamic therapy (PDT) and clofibric acid (CA) to treat peritoneal carcinomatosis resulting from ovarian cancer. Materials and Methods: Nude rats were used to create a model of peritoneal carcinomatosis resulting from ovarian cancer and the effectiveness of PDT with 5-aminolevulinic acid methyl ester hydrochloride (methyl-ALA-PDT) was determined. The survival time of rats receiving that therapy was compared to the survival time of a control group. Rats with peritoneal carcinomatosis resulting from ovarian cancer were divided into 3 groups: a group that received debulking surgery (DS) alone, a group that received DS+methyl-ALA-PDT, and a group that received DS+methyl-ALA-PDT+CA. The survival time of the 3 groups was compared. Protoporphyrin, a metabolite of methyl-ALA, produces a photochemical action when activated by light. The level of protoporphyrin (the concentration) that reached organs in the abdomen was measured with HPLC. Results: Rats receiving methyl-ALA-PDT had a significantly longer survival time compared to the controls. Rats with peritoneal carcinomatosis that received DS+methyl-ALA-PDT+CA had a significantly longer survival time compared to the rats that received DS alone. Some of the rats that received concurrent therapy survived for a prolonged period. Protoporphyrin was highly concentrated in peritoneal metastases, but only small amounts reached major organs in the abdomen. PDT was not found to result in necrosis in the intestines. Conclusions: The results indicated that concurrent therapy consisting of PDT with methyl-ALA and CA is effective at treating peritoneal carcinomatosis resulting from ovarian cancer without damaging organs.

Effects of ${\alpha}$-lipoic acid and L-carnosine supplementation on antioxidant activities and lipid profiles in rats

  • Kim, Mi-Young;Kim, Eun-Jin;Kim, Young-Nam;Choi, Chang-Sun;Lee, Bog-Hieu
    • Nutrition Research and Practice
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    • v.5 no.5
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    • pp.421-428
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    • 2011
  • ${\alpha}$-Lipoic acid and L-carnosine are powerful antioxidants and are often used as a health supplement and as an ergogenic aid. The objective of this study was to investigate the effects of ${\alpha}$-lipoic acid and/or L-carnosine supplementation on antioxidant activity in serum, skin, and liver of rats and blood lipid profiles for 6 weeks. Four treatment groups received diets containing regular rat chow diet (control, CON), 0.5% ${\alpha}$-lipoic acid (ALA), 0.25% ${\alpha}$-lipoic acid+0.25% L-carnosine (ALA+LC), or 0.5% L-carnosine (LC). Superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) and lipid peroxidation products, malondialdehyde (MDA) concentrations, were analyzed in serum, skin, and liver. Blood lipid profiles were measured, including triglycerides (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C). Skin and liver SOD activities of the ALA and LC groups were higher than those of the CON group (P<0.05), but serum SOD activity was higher only in the LC group compared to that in the CON group (P<0.05). Additionally, only liver GSH-Px activity in the LC group was higher than that of the CON and the other groups. Serum and skin MDA levels in the ALA and LC groups were lower than those in the CON group (P<0.05). Serum TG and TC in the ALA and ALA+LC groups were lower than those in the CON and LC groups (P<0.05). The HDL-C level in the LC group was higher than that in any other group (P<0.05). LDL-C level was lower in the ALA+LC and LC groups than that in the CON group (P<0.05). Thus, ${\alpha}$-lipoic acid and L-carnosine supplementation increased antioxidant activity, decreased lipid peroxidation in the serum, liver, and skin of rats and positively modified blood lipid profiles.

ADPRT Val762Ala and XRCC1 Arg194Trp Polymorphisms and Risk of Gastric Cancer in Sichuan of China

  • Wen, Yuan-Yuan;Pan, Xiong-Fei;Loh, Marie;Tian, Zhi;Yang, Shu-Juan;Lv, Si-Han;Huang, Wen-Zhi;Huang, He;Xie, Yao;Soong, Richie;Yang, Chun-Xia
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2139-2144
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    • 2012
  • Objective: Gastric cancer remains a major health problem in China. We hypothesized that XRCC1 Arg194Trp and ADPRT Val762Ala may be associated with risk. Methods: We designed a multicenter 1:1 matched case-control study of 307 pairs of gastric cancers and controls between October 2010 and August 2011. XRCC1 Arg194Trp and ADPRT Val762Ala were sequenced, and demographic data as well as lifestyle factors were collected using a self-designed questionnaire. Results: Individuals carrying XRCC1 Trp/Trp or Arg/Trp variant genotype had a significantly increased risk of gastric cancer (OR, 1.718; 95% CI, 1.190-2.479), while the OR for ADPRT Val762Ala variant genotype (Ala/Ala or Val/Ala) was 1.175 (95% CI, 0.796-1.737). No gene-gene or gene-environment interactions were found. In addition, family history of cancer and drinkers proportion were higher among cases than among controls (P<0.05). Conclusions: XRCC1 194 Arg/Trp or Trp/Trp genotype, family history of cancer, and drinking are suspected risk factors of gastric cancer from our study. Our findings may offer insight into further similar large gene-environment and gene-gene studies in this region.