• Title/Summary/Keyword: 5-Aminolevulinic Acid

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Porphyrin Derivatives from a Recombinant Escherichia coli Grown on Chemically Defined Medium

  • Lee, Min Ju;Chun, Se-Jin;Kim, Hye-Jung;Kwon, An Sung;Jun, Soo Youn;Kang, Sang Hyeon;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.22 no.12
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    • pp.1653-1658
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    • 2012
  • We have reported previously that a recombinant Escherichia coli co-expresses aminolevulinic acid (ALA) synthase, an NADP-dependent malic enzyme, and a dicarboxylate transporter-produced heme, an iron-chelated porphyrin, in a succinate-containing complex medium. To develop an industrially plausible process, a chemically defined medium was formulated based on M9 minimal medium. Heme synthesis was enhanced by adding sodium bicarbonate, which strengthened the C4 metabolism required for the precursor metabolite, although a pH change discouraged cell growth. Increasing the medium pH buffering capacity (100mM phosphate buffer) and adding sodium bicarbonate enabled the recombinant E. coli to produce heme at rates 60% greater than those in M9 minimal medium. Adding growth factors (1 mg/l thiamin, 0.01 mg/l biotin, 5 mg/l nicotinic acid, 1 mg/l pantothenic acid, and 1.4 mg/l cobalamin) also induced positive heme production effects at levels twice of heme production in M9-based medium. Porphyrin derivatives and heme were found in the chemically defined medium, and their presence was confirmed by liquid chromatography/mass spectroscopy (LC/MS). The formulated medium allowed for the production of $0.6{\mu}M$ heme, $29{\mu}M$ ALA, $0.07{\mu}M$ coproporphyrin I, $0.21{\mu}M$ coproporphyrin III, and $0.23{\mu}M$ uroporphyrin in a 3 L pH-controlled culture.

A Spectral Comparison Study of PDT Drugs - ALA and ALA-Hexyl ester

  • Yao, Chen-Ji;Peng Qian;JodI H-J
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.521-523
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    • 2002
  • 5-aminolevulinic acid (ALA) has been used to stimulate endogenous protoporphyrin IX (PpIX) in tumor and then initiate PDT. Recently, ALA-Hexyl ester (He-ALA) was found much effective than ALA on producing PpIX in cancer cells. To clarify the transportation mechanism of ALA and He-ALA, the detection of them is the important step. ALA and its derivatives all don't emit fluorescence, so the Raman spectroscopy was used here for the direct detection of ALA and He-ALA. The results showed that ALA and He-ALA have the common strong Raman peaks at 2930, 2950 CM$\^$-1/, due to the CH$_2$ vibration. The peak 3050 CM$\^$-1/ appeared in ALA spectrum can be attributed to OH vibration, while the peaks of 2860, 2900 CM$\^$-1/ in He-ALA spectrum were assigned as the modes of CH$_3$. This Raman spectral characteristic is consistence with the structure difference of He-ALA and ALA. Thus, Raman spectroscopy provides a new way to detect and distinguish ALA and He-ALA, and could be explored further in biology system.

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Effects of Na-alginate on the Toxicity Induced by Lead and Cadmium in Albino Rats (Na-alginate 투여가 白鼠의 鉛과 카드뮴 독성에 미치는 영향)

  • Lee, Sang Bong
    • Journal of Environmental Health Sciences
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    • v.12 no.2
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    • pp.17-26
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    • 1986
  • Earlier studies have shown that the administration of Na-alginate, a water-soluble, non absorbable acidic polysaccharide, effectively reduces the absorption of radioactive strontium, cadmium and other bivalent metals. In this study the effects of Na-alginate on the Toxicity induced by Lead and Cadmium were examined in rats and the following results were obtained. 1. The elevations of the activities of serum GOT, GPT by Pb and Cd administration in the experimental animals. However, the administration of Na-alginate significantly inhibited the elevation of serum GOT, GPT. 2. Generally serum TBA values were significantly elevated in Pb and Cd group in comparison with the control group. However, the administration of Na-alginate inhibited the elevation of serum TBA values. 3. The activity of Alkaline phosphatase was also significantly elevated in Cd group in comparison with the control group. However, the administration of Na-alginate significantly inhibited the elevation of serum alkaline phsphatase. 4. Furthermore, the activity of $\delta$-aminolevulinic acid dehydratase in red blood cells was also significantly inhibited by Pb and this inhibition was stimulated by the administration of Na-alginate. 5. The concentrations of Pb in liver and Cd in kidney were markedly increased by Pb and Cd. However, the administration of Na-alginate significantly inhibited the elevation of them.

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Hematological Characterization of Penaeus chinensis by Feed-additives (사료 첨가제에 의한 대하의 혈액학적 특성)

  • Kim, Yeong-Jin;Lee, Sun-Yi;Cho, Hyo-Jin;You, Sun-Nyoung;Kim, Kwang-Youn;Choi, Won-Chul;Heo, Moon-Soo;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1087-1089
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    • 2006
  • Shrimp has efficient non-specific defense mechanisms based on activities of the hemocytes against pathogens. Up to now, it has been known that one of the non-specific immune reactions is related to mutual association among types of hemocyte, granular cell, semi-granular cell, and hyaline cell. In this study, we tried to know the effectiveness of feed-additives such as 5-aminolevulinic acid(ALA), chitosan, and hot-water extract of herb on immunity of shrimp (Penaeus chinensis) by hemocytic observation and SDS-PAGE analysis. Finally, we suggest a principle of the examination system for effects of various feed-additives.

A Multi-detection Fluorescence Dye with 5-ALA and ICG Using Modified Light Emitting Diodes

  • Yoon, Kicheol;Kim, Eunji;Kim, Kwanggi;Lee, Seunghoon;Yoo, Heon
    • Current Optics and Photonics
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    • v.3 no.3
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    • pp.256-262
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    • 2019
  • Extensive tumor resection accompanied by radiotherapy and chemotherapy is the standard of care for malignant gliomas. However, there is a significant obstacle to the complete resection of the tumor due to the difficulty of distinguishing tumor and normal brain tissue with a conventional surgical microscope. Recently, multiple studies have shown the possibility of fluorescence-guided surgery in malignant gliomas. The most used fluorescence dyes for brain tumor surgery are 5-aminolevulinic acid (5-ALA) and indocyanine green (ICG). In this paper, a new fluorescence guided operation system, which can detect both 5-ALA and ICG fluorescent images simultaneously, is presented. This operation system consists of light emitting diodes (LEDs) which emits 410 nm and 740 nm wavelengths. We have performed experiments on rats in order to verify the operation of the newly developed operation system. Oral administration and imaging were performed to observe the fluorescence of 5-ALA and ICG fluorescence in rats. When LEDs at wavelengths of 410 nm and 740 nm were irradiated on rats, 628 nm wavelength with a violet fluorescence color and 825 nm wavelength with a red fluorescence color were expressed in 5-ALA and ICG fluorescent material, respectively, thus we were able to distinguish the tumor tissues easily. Previously, due to the poor resolution of the conventional surgical microscope and the fact that the color of the vein is similar to that of the tumor, the tumor resection margin was not easy to observe, thus increasing the likelihood for cancer recurrence. However, when the tumor is observed through the fluorescence guided operation system, it is possible to easily distinguish the color with the naked eye and it can be completely removed. Therefore, it is expected that surgical removal of cancerous tumors will be possible and surgical applications and surgical microscopes for cancer tumor removal surgery will be promising in the future.

Effect of Promoters on the Heme Production in a Recombinant Corynebacterium glutamicum (재조합 Corynebacterium glutamicum으로부터 헴첼 생산에 미치는 프로모터의 효과)

  • Yang, Hyungmo;Kim, Pil
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.337-342
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    • 2019
  • We published that bacterial heme was over-produced in a recombinant Corynebacterium glutamicum expressing 5-aminolevulinic acid synthase ($hemA^+$) under control of a constitutive promoter ($P_{180}$) and the heme-producing C. glutamicum had commercial potentials; as an iron feed additive for swine and as a preservative for lactic acid bacteria. To enhance the heme production, the $hemA^+$ gene was expressed under controls of various promoters in the recombinant C. glutamicum. The $hemA^+$ expression by $P_{gapA}$ (a constitutive glycolytic promoter of glyceraldehyde-3-phosphate dehydrogenase) led 75% increase of heme production while the expression by $P_{H36}$ (a constitutive, very strong synthetic promoter) resulted in 50% decrease compared with the control ($hemA^+$ expression by $P_{180}$ constitutive promoter). The $hemA^+$ expression by a late log-phase activating $P_{sod}$ (an oxidative-stress responding promoter of superoxide dismutase) led 50% greater heme production than the control. The $hemA^+$ expression led by a heat-shock responding chaperone promoter ($P_{dnaK}$) resulted in 121% increase of heme production at the optimized heat-shock conditions. The promoter strength and induction phase are discussed based on the results for the heme production at an industrial scale.

Study on the Biosynthetic Characteristics of Photosynthetic Pigments in Dodder(Cuscuta australis R. Br.) Plant (실새삼의 광합성색소 생합성특성에 관한 연구)

  • Kim, J.S.;Kwak, H.H.;Kim, B.C.;Cho, K.Y.
    • Korean Journal of Weed Science
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    • v.17 no.3
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    • pp.314-324
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    • 1997
  • This study was conducted to investigate whether 1) photosynthetic pigments(chlorophylls and carotenoids) are formed in dodder plant(Cuscuta australis), 2) there are any characteristics in the pigment biosynthesis, compared to that of other normal plants, and 3) dodder responds to some herbicides having target site on chloroplast. 1. Chlorophyll content of dodder tendrill grown under a natural daylight was 9 times and 50 times lower than that of field bindweed stem and leaf, respectively. 2. The photosynthetic pigment contents varied in different tissues, being higher in a apical region than in a lower region of seedling or tendrill. Chlorophyll wasn't almost observed below the 4th internode from the upper. 3. Pigment contents were greatly dependent on light intensity so that there were 4 to 6 times difference among light conditions. When the shoot containning low pigment contents under natural light, was incubated in growth chamber with various light intensities, the pigment contents were increased by 3 times of initial contents at about 97${\mu}E$ $m^{-2}s^{-1}$PAR. While the change in pigment contents was not observed at above 450${\mu}E$ $m^{-2}s^{-1}$PAR 4. Exogenous supply of 5mM 5-aminolevulinic acid increased protochlorophyllide by 7 times and 1.4 times in the etiolated shoot from field bindweed rhizome and in dodder stem, respectively, showing that dodder relatively has a low response to 5-aminolevulinic acid. 5. Pigment loss was observed in the treatment of paraquat, norflurazon, oxyfluorfen and diuron, and protoporphyrin IX was accumulated by oxyfluorfen as in normal plants Based on above results, several chracteristics of pigment biosynthesis in dodder seem to be summerized as follows. Photosynthetic pigment biosynthesis in Cuscuta australis runs even in low level. The pigment contents is differentially distributed in different regions and their contents seem significantly to be controlled by light intensities. Especially, chloroplast rapidly tends to degenerate with the development of tissue. Some herbicides having target site on chloroplast induce damage to dodder stem but are unlikely to control it well in field, except paraquat, due to low chloroplast activity and parasitic mode of nutrition.

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Effects of Dietary Vitamin E and Selenium on Hematopoiesis and Antioxidative Detoxification Mechanism in Lead Poisoned Rats (식이 Vitamin E와 Selenium이 납중독된 흰쥐에 있어서 조혈작용과 항산화적 해독기구에 미치는 영향)

  • 이순재;박규영;김관유
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.6
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    • pp.651-657
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    • 1993
  • The protective effects of dietary vatamin E and selenium on peroxidative damage and hematopoietic inhibition by lead poisoning were investigated in rats. Male Sprague-Dawley rats weighing 150$\pm$5g were divided into six groups according to dietary vitamin E and / or selenium levels, i.e. control(vitamin E, 40mg/kg diet), 0E(without vitamin E, Se), 40E(vitamin E, 40mg/kg diet ; without Se), 200E(vitamin E, 200mg/kg diet ; without Se), 200ES(vitamin E, 200mg/kg diet ; Se, 0.5ppm) and 0Es(without vitamin E ; Se, 0.5ppm) groups. All experimental groups were fed ad libitum 2000ppm lead in diet except control for 4 weeks. Hemoglobin contents and hematocrit values of lead groups were lower than control group except 200ES group and were the lowest in 0E group. Aminolevulinic acid dehydratase(ALAD) activities of blood and liver were sequentially reduced in 200ES, 200E, 0ES, 40E and 0E groups, compared to control, were as urinary aminolevulinic acid (ALA) excretions were increased in the groups which represented low ALAD activity. Heapatic superoxide dismutase(SOD) activities was lower in 0E, and higher in 40E, 200E and 200ES groups, compared with control. Glutathione peroxidase(GPX) activities of liver were reduced in 0E and 40E groups, but those of 0ES, 200E and 200ES groups were significantly increased. Especially GPX activities in 200ES and 200ES groups were not different from control group. The reduced glutathione contents in liver were lowest in 0E and 40E groups, compared with control, whereas levels of the oxidized form were opposite phenomena of that. Liver lipid peroxide values of 0E, 0ES, 40E and 200E groups were 6.4, 2.9, 2.1 and 1.3 fold higher than control, respectively, but 200ES groups was not different from control.

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The Effect of Occupational Health Service on the Improvement of Worker’s Health in a Lead Using Industry (모연취급 사업장에서의 산업보건사업이 근로자 건강증진에 미치는 효과)

  • Lee, Byeong-Guk;Lee, Gwang-Muk;An, Gyu-Dong
    • 산업보건소식
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    • no.46
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    • pp.3-13
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    • 1987
  • For the purpose of investigating the effect of occupational health service in terms of environmental control and health provision of workers on the improvement of worker's health, authors analysed the data of environmental measurement and health check-up of one lead using industry who started his investment to environmental improvement from 1980. Six hundred million won was invested for environmental improvement from 1980 to 1986. This investment brought about apparent improvement of working conditions of all of the workplaces from mean concentration of lead in air over 0.15mg/$m^3$ 1981 to mean concentration of lead in air less than 0.15mg/$m^3$. Environmental control reduced mean blood lead level from 51.2 $\pm$ 11.5ug/이 in 1983 to 39.2 $\pm$ 16.0ug/dl in 1986, and delta-aminolevulinic acid concentration 3.15 $\pm$ 2.1mg/l in 1982 to 1.96 $\pm$ 1. 7mg/l in 1986, respectively. Blood ZPP levels were decreased from 76.1 $\pm$ 58.9ug/dl in 1983 to 42.23 $\pm$ 30.3ug/dl in 1986. If 150ug/dl of blood ZPP is considered as unacceptable limit of lead intoxication, more than 10 percent of workers belonged to this category in 1983, but only 0.5 of workers showed their blood ZPP level over the 150ug/dl in 1986 It was observed that no workers whose work duration were less than one year showed their blood ZPP level over 100ug/dl, but there were high percentages of lead workers with high blood ZPP (over 150 ug/dl) who were working in uncontrolled had working condition and whose work duration were less than one year.

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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.