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$HgCl_2$ Dysregulates the Immune Response of Balb/c Mice (수은에 의한 마우스의 면역반응 조절장애)

  • Ki, No-Suk;Koh, Dai-Ha;Kim, Chong-Suh;Lee, Jung-Sang;Kim, Nam-Song;Lee, Hwang-Ho
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.1 s.45
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    • pp.11-24
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    • 1994
  • The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

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Regulation of Gb3 Expression on Dendritic Cells (수지상세포에 있어서 베로독소 수용체의 발현조절)

  • Lim, Suk-Hwan;Kim, Gi-Young;Kim, Hyung-Chun;Kim, Young-Hee;Son, Yong-Hae;Oh, Yang-Hyo;Park, Yeong-Min
    • Journal of Life Science
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    • v.17 no.4 s.84
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    • pp.482-492
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    • 2007
  • Infection with Shiga-like toxin (SLT)-producing Escherichia coli causes a spectrum of illnesses with high morbidity and mortality. Host mediators play an important role in the pathogenesis of SLT-I toxicity. We here investigated the effect of SLT-I on tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ production, effect of $TNF-{\alpha}$ on glycolipid globotriaosyleramide (Gb3) expression, and relationship between Gb3 level and differential susceptibility of cells to SLT-I. In this study, we observed that detectable levels of $TNF-{\alpha}$ are produced 6 hrs after induction and continued to increase during 48 hrs by SLT-I. It was also found that Vero cells and dendritic cells expressed high levels of Gb3, 83% and 68%, respectively, and that macrophages had a low level of Gb3 (29%) and showed refractory to cytotoxicity against SLT-I. Vero cells and dendritic cells expressing high levels of Gb3 were highly susceptible to SLT-I. furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. These results suggest that the expression of Gb3 is necessary, but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its receptor, Gb3, in the pathogenesis of E. coli O157 infection.

Inhibitory Effects of Asparagus cochinchinensis in LPS-Stimulated BV-2 Microglial Cells through Regulation of Neuroinflammatory Mediators, the MAP Kinase Pathway, and the Cell Cycle (Lipopolysaccharide로 자극된 BV-2 미세교세포에서 신경염증 매개체, MAP kinase경로, 세포주기의 조절에 의한 천문동(Asparagus cochinchinensis)의 저해효과)

  • Lee, Hyun Ah;Kim, Ji Eun;Choi, Jun Young;Sung, Ji Eun;Youn, Woo Bin;Son, Hong Joo;Lee, Hee Seob;Kang, Hyun-Gu;Hwang, Dae Youn
    • Journal of Life Science
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    • v.30 no.4
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    • pp.331-342
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    • 2020
  • The suppression of neuroinflammatory responses in microglial cells can be considered a key target for improving the progression of neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). Asparagus cochinchinensis has traditionally been used as a medicine to treat fever, cough, kidney disease, breast cancer, inflammatory diseases, and brain diseases. In this study, we investigated the neuroprotective mechanism of an aqueous extract from A. cochinchinensis root (AEAC), particularly its anti-inflammatory effects on lipopolysaccharide (LPS)-activated BV-2 microglial cells. BV-2 cells were treated with four different concentrations of AEAC. No significant toxicity was detected in BV-2 cells treated with AEAC. Nitric oxide (NO), cyclooxygenase-2 (COX-2) mRNA, and inducible nitric oxide synthase (iNOS) mRNA levels were 21% lower in the AEAC+LPS group than in the Vehicle+LPS group. Lower proinflammatory (TNF-α and IL-1β) and anti-inflammatory cytokine (IL-6 and IL-10) levels were also detected in the AEAC+LPS group than in the Vehicle+LPS group, albeit at varying rates. Moreover, the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group compared to the Vehicle+LPS group, enhancement of the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group, while cell cycle arrest at the G2/M phase caused by LPS treatment was less severe in the AEAC+LPS group. The increase in reactive oxygen species (ROS) generation induced by LPS treatment was also lower in the AEAC-pretreated group than in the Vehicle+LPS group. This is the first study to show that AEAC exerts anti-neuroinflammatory activity against LPS stimulation by regulating the MAPK signaling pathway, the cell cycle, and ROS production.

Purification of Vibrio anguillarum Growth Inhibition Factor Produced by Bacillus amyloliquefaciens H41. (Bacillus amyloliquefaciens H41이 생산하는 Vibrio anguillarum 생육 저해인자의 정제)

  • Shin, Hyun-Chul;Chung, Kyung-Tae;Kim, Kwang-Hyun;Kim, Byung-Woo;Kwon, Hyun-Ju;Lee, Eun-Woo;Yum, Jong-Hwa;Rhu, Eun-Ju;Jeong, Yu-Jeong;Kim, Young-Hee
    • Journal of Life Science
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    • v.18 no.6
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    • pp.789-795
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    • 2008
  • To study the possible use of probiotics in fish farming, we evaluated antagonism of antibacterial strain Bacillus amyloliquefaciens H41 against the fish pathogenic bacterium Vibrio anguillarum NCMB1. The purification of growth inhibition factor produced by B. amyloliquefaciens H41 was achieved by obtaining supernatant of this bacterium. The growth inhibition factor was purified to homogeneity by 70% ammonium sulfate precipitation, DEAE-sephadex A-50 ion exchange chromatography, sephadex G-200 gel filtration column chromatography, and sephadex G-50 gel filtration column chromatography with 40.8 fold of purification and 2.9% yield. The molecular weight of the purified growth inhibition factor was 48 kDa as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The optimum pH and temperature for the growth inhibition factor were pH 7.5 and $30^{\circ}C$, respectively. The activity of growth inhibition factor was enhanced slightly by some metal ions, such as $Mg^{+2}$, $Mn^{+2}$, but was inhibited by the addition of $Co^{+2}$, $Hg^{+2}$, $Zn^{+2}$ and $Ag^{+2}$. NaCl stability of the growth inhibition factor was observed with 50% residual activity at 3% NaCl concentration. Toxicity test showed that the purified B. amyloliquefaciens H41 growth inhibition factor did not affect the live of Japanese flounder (Paralichthys olivaceus) and the effectiveness was 78% of residual lethality compared to commercial antibacterial agents.

The Anticancer Effect and Mechanism of Photodynamic Therapy Using 9-Hydroxypheophorbide-a and 660 nm Diode Laser on Human Squamous Carcinoma Cell Line. (9-hydroxypheophorbide-a와 660 nm 다이오드 레이저를 이용한 광역학치료의 항암효과와 치료기전에 대한 연구)

  • Ahn, Jin-Chul
    • Journal of Life Science
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    • v.19 no.6
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    • pp.770-780
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    • 2009
  • A new photosensitizer, 9-Hydroxypheophorbide-a (9-HpbD-a), was derived from Spirulina platensis. We conducted a series of experiments, in vitro and in vivo, to evaluate the anticancer effect and mechanism of photodynamic therapy using 9-HpbD-a and 660 nm diode lasers on a squamous carcinoma cell line. We studied the cytotoxic effects of pheophytin-a, 9-HpbD-a, 9-HpbD-a red and 660 nm diode lasers in a human head and neck cancer cell line (SNU-1041). Cell growth inhibition was determined by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay. The effects of 9-HpbD was higher than those of 9-HpbD-a red or pheophytin-a in PDT. We then tested the cytotoxic effects of 9-hydroxypheophorbide-a (9-HpbD-a) in vitro. The cultured SNU-I041 cells were treated with serial concentrations of 9-HpbD-a followed by various energy doses (0, 0.1, 0.5, 3.2 J/$cm^{2}$) and by various interval times (0, 3, 6, 9, 12 hr) until laser irradiation, then MTT assay was applied to measure the relative inhibitory effects of photodynamic therapy (PDT). Optimal laser irradiation time was 30 minutes and the cytotoxic effects according to incubation time after 9-HpbD-a treatment increased until 6 hours, after which it then showed no increase. To observe the cell death mechanism after PDT, SUN-I041 cells were stained by Hoechst 33342 and propidium iodide after PDT, and observed under transmission electron microscopy (TEM). The principal mechanism of PDT at a low dose of 9-HpbD-a was apoptosis, and at a high dose of 9-HpbD-a it was necrosis. PDT effects were also observed in a xenografted nude mouse model. Group I (no 9-HpbD-a, no laser irradiation) and Group II (9-HpbD-a injection only) showed no response (4/4, 100%), and Group III (laser irradiation only) showed recurrence (1/4,25%) or no response (3/4, 75 %). Group IV (9-HpbD-a + laser irradiation) showed complete response (10/16, 62.5%), recurrence (4/16, 25%) or no response (2/16, 12.5%). Group IV showed a significant remission rate compared to other groups (p<0.05). These results suggest that 9-HpbD-a is a promising photosensitizer for the future and that further studies on biodistribution, toxicity and mechanism of action would be needed to use 9-HpbD-a as a photosensitizer in the clinical setting.

Degradation of the Herbicide, TOK(2,4-dichloro-4'-nitro diphenyl ether) in Soil (제초제(除草劑) TOK의 토양중(土壤中) 분해(分解))

  • Lee, Jae-Koo;Kim, Ki-Cheol;Kim, Chang-Han
    • Applied Biological Chemistry
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    • v.23 no.2
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    • pp.131-139
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    • 1980
  • TOK (2,4-Dichlolo-4'-nitrodiphenyl ether) was applied to two Korean soils possessing different physico-chemical properties at a certain concentration and incubated for a certain time under flooded conditions. The metabolites and the soil microorganisms involved in the degradation of TOK are studied. Chong Ju and Chung Ju soils treated with TOK at a concentration of 500 ppm and incubated for two, four, and six months at $30^{\circ}C$ yielded 4-chloro-4'-amino diphenyl ether, 2, 4-dichloro-4'-amino diphenyl ether(amino-TOK), N-[4'-(4-chloro-phenoxy)] phenyl acetamide, and N-[4'-(4-chloro-phenoxy)] phenyl formamide as the major metabolites. TOK underwent the reduction of nitrogroup to amino group, dechlorination, acetylation, and formylation. No cleavage at the ether linkage was recognized. TOK was more readily degraded in Chung Ju soil which is characterized by the higher pH (PH 6.43), clay loam in textural class, and the higher cation exchange capacity. The toxicity of TOK as a possible environmental contaminant is expected to be considerably reduced as a result of the above degradation Twelve strains of soil bacteria were isolated from the TOK-treated Chong Ju and Chung Ju soils. As a result of the incubation of TOK in pure cultures of the isolates, T-1-1 strain isolated from Chong Ju soil had almost no degradability, whereas T-2-3 strain turned out to be the most potent. The degradation of TOK by the isolates constituted mostly the reduction of the nitro group to amino group. The citrate buffer extract of Chung Ju soil reduced TOK more readily to amino-TOK than that of Chong Ju soil.

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Degradation Kinetic and Mechanism of Methyl Tert-butyl Ether (MTBE) by the Modified Photo-Fenton Reaction (Modified Photo-Fenton Reaction을 이용한 Methyl Tert-butyl Ether (MTBE)의 분해 Kinetic 및 메커니즘 규명에 관한 연구)

  • Kim, Min-Kyoung;Kong, Sung-Ho
    • Journal of Soil and Groundwater Environment
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    • v.11 no.6
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    • pp.69-75
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    • 2006
  • Improper disposal of petroleum and spills from underground storage tanks have created large areas with highly toxic contamination of the soil and groundwater. Methyl tert-butyl ether (MTBE) is widely used as a fuel additive because of its advantageous properties of increasing the octane value and reducing carbon monoxide and hydrocarbon exhausts. However, MTBE is categorized as a possible human carcinogen. This research investigated the Modified Photo-Fenton system which is based on the Modified Fenton reaction and UV light irradiation. The Modified Fenton reaction is effective for MTBE degradation near a neutral pH, using the ferric ion complex composed of a ferric ion and environmentally friendly organic chelating agents. This research was intended to treat high concentrations of MTBE; thus, 1,000 mg/L MTBE was chosen. The objectives of this research are to find the optimal reaction conditions and to elucidate the kinetic and mechanism of MTBE degradation by the Modified Photo-Fenton reaction. Based on the results of experiments, citrate was chosen among eight chelating agents as the candidate for the Modified Photo-Fenton reaction because it has a relatively higher final pH and MTBE removal efficiency than the others, and it has a relatively low toxicity and is rapidly biodegradable. MTBE degradation was found to follow pseudo-first-order kinetics. Under the optimum conditions, [$Fe^{3+}$] : [Citrate] = 1 mM: 4 mM, 3% $H_2O_2$, 17.4 kWh/L UV dose, and initial pH 6.0, the 1000 ppm MTBE was degraded by 86.75% within 6 hours and 99.99% within 16 hours. The final pH value was 6.02. The degradation mechanism of MTBE by the Modified Photo-Fenton Reaction included two diverse pathways and tert-butyl formate (TBF) was identified to be the major degradation intermediate. Attributed to the high solubility, stability, and reactivity of the ferric-citrate complexes in the near neutral condition, this Modified Photo-Fenton reaction is a promising treatment process for high concentrations of MTBE under or near a neutral pH.

Influence of Toxic Heavy Metals on Germination of Rice Seeds and Growth of Rice Seedling (수도생육(水稻生育)에 대한 유해(有害) 중금속(重金屬)의 영향(影響) - 발아 및 묘대기(苗垈期) 생육(生育)에 대하여 -)

  • Kim, B.J.;Ha, Y.L.;Kim, J.O.;Han, K.H.
    • Korean Journal of Soil Science and Fertilizer
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    • v.11 no.2
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    • pp.119-126
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    • 1979
  • Rice seeds, suweon 264, were germinated under 5 levels of toxic heavy metals, Cd(0, 0.05, 0.5, 5, 20ppm), Cu(0, 0.05, 0.5, 5, 20, ppm), Cr(0, 0.5, 1, 5, 10ppm), Ni(0, 0.5, 1, 5, 10ppm), Co(0, 0.5, 1, 5, 10ppm), Zn(0, 0.5, 5, 20, 40, ppm), Pb(0, 0.5, 5, 20, 40ppm) and Mn(0, 1, 10, 25, 50, ppm) in culture solution, and then grown with supplying culture solution contained respective concentrations. Germination and growth response to the toxic heavy metals were studied. Results obtained are as follows : 1) The germination injury of rice seeds by excess concentration of toxic heave metal in culture solution occured in Cd and Cu; below 0.05 ppm, Ni; below 0.5 ppm, Mn; below 1.0 ppm, Co and Cr; 0.5-1.0 ppm, and 0.5-5 ppm, Zn and Pb. Thereby, in the order of degrees of the elements toxicity to germination, they were arranged as follows : Cd>Cu>Ni>Co>Cr>Mn>Zn$$\geq_-$$Pb. 2) Toxic heavy metal concentrations in culture solution, which result in decreasing dry weight due to the injury of excess concentration of treated elements, were Cd: below 0.05 ppm, Ni, Cr and Co; below 0.5 ppm, Cu and Zn; 0.5-5 ppm, Pb; 5-20 ppm and Mn; 10-25 ppm. The order was Ni>Cd>Cr>Co>Cu$$\geq_-$$Zn>Pb>Mn. 3) The critical contents of Cd, Ni, Pb, Cu, Zn, Mn, and Co in dry matter, Which result in decreasing dry weight, were considered to be 0.05-15.5, 1.50-25.0, 24.0-28.0, 26.5-62.5, 470-645.0, 231.0-500.0 and below 15.0 ppm, respectively. 4) The contents of Cr, Co, Cd, Pb, Ni, Cu and Zn in dry matter by 0.5 ppm treatment concentration of each heavy metals was trace, 15.0, 17.5, 24.0, 25.0, 84.5 and 470.0 ppm, respectively. Thereby, in the order of each element to uptaked by rice seedlings, they were arranged as follow; Zn>Cu>Ni>Pb>Cd>Co>Cr. 5) The hazardous concentrations of root activity by toxic heavy metals in culture solution were Cd; below 0.05, Cu; 0.05-0.5, Cr; below 0.5, Ni; 0.5-1.0, Co; 0.5-1.0, Zn; above 0.5, Pb; 0.5-5.0 and Mn; 1.0-10.0 ppm. The hazardous degree of root activity by toxic heave metals was in the order of Cd>Cu>Cr>Zn>Ni>Co>Pb>Mn.

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Determination of Sulfur Requirement to Adjust pH of Alkaline Soil by Buffer Curve Method (알칼리성 토양 pH 교정시 완충곡선법을 이용한 황 시용량 결정)

  • Lee, In-Bog;Lim, Jae-Hyun;Yiem, Myoung-Soon
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.6
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    • pp.405-415
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    • 2000
  • To determine application rate of elemental sulfur to adjust pH of alkaline soil, buffer curve method was investigated. The elemental sulfur required to control pH 8.3 to pH 6.3 by buffer curve calculation was treated in two soils of silty loam and sandy loam, and the sulfur-mixed soils were moistened with 50% of water holding capacity during incubation of 6 weeks at $30^{\circ}C$. Soil pH was lowered with incubation and reached to target point after 4 weeks of incubation, and elemental sulfur was oxidised entirely to sulfate. This means that buffer curve has the accuracy to determine sulfur application rate in alkaline soil. However it is estimated that application rate of sulfur should be carefully determined in the field scale. Excess application of elemental sulfur resulted in extremely low soil pH and caused the hinderance of lettuce growth by nutritional imbalance and ion toxicity. To simplify the determination procedure of sulfur requirement, buffer curve method by addition of 0.1N-HCl solution as unit of mL was developed, it was compared with theroutine methods which diluted $H_2SO_4$ solution and $Ca(OH)_2$ are added as cmolc per kg soil to adjust each pH step. Buffer capacities, cmolc kg $soil^{-1}$ $pH^{-1}$, calculated from two buffer curves were not significantly different. The result indicates that buffer curve method by 0.1N-HCl can be used to adjust high pH of alkaline soil.

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Long-term Results of Radiotherapy for Subfoveal Choroidal Neovascularization (CNV) in Age-related Macular Degeneration (ARMD) (연령관련 황반하 맥락막 신생혈관증에서 방사선의 장기적 치료 결과)

  • Kim, Bo-Kyong;Kim, Il-Han
    • Radiation Oncology Journal
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    • v.25 no.2
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    • pp.63-69
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    • 2007
  • [ $\underline{Purpose}$ ]: We performed this prospective randomized study to evaluate the efficacy and the complications of radiotherapy for Subfoveal CNV in ARMD and to compare the treatment results at two dosages (14.4 Gy and 19.8 Gy). $\underline{Materials\;and\;Methods}$: 60 eyes of 55 patients were enrolled, and randomized into 14.4 Gy (31 eyes) or 19.8 Gy (29 eyes) groups. CT was used to plan the radiotherapy. All patients received radiotherapy with a 1.8 Gy daily dose using 4 MV photon. We categorized treatment results as improved, stable, or deteriorated based on visual acuity changes of more than 2 lines on the ETDRS chart. $\underline{Results}$: Median follow-up period was 33.5 months. At 12 months, visual acuity improved in 9 (16.7%), stable in 41 (75.9%), and aggravated in 4 (7.4%) of 54 evaluated eyes. At 24 months, 49 eyes (81.7%) were evaluated. Visual acuity improved in 6 (12.2%), was stable in 33 (67.4%), and deteriorated in 10 (20.4%). At 36 months, 37 eyes were evaluated. Six (16.2%) eyes were improved, 21 (56.8%) stable, and 10 (27.0%) deteriorated. No significant difference in response was observed between the 14.4 Gy and 19.8 Gy groups (Mantel-Haenszel $x^2=0.4756$). The proportion of eyes with a vision of $20/100{\leq}increased$ from 28.3% initially to 32.7% after 24 months of radiotherapy. There were no severe acute or chronic complications. $\underline{Conclusion}$: External beam radiotherapy with doses of 14.4 or 19.8 Gy may be an effective treatment for subfoveal CNV in ARMD. No dose-response relationships with respect to treatment response or toxicity were observed between the 14.4 Gy and 19.8 Gy groups.