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A Gelatinase A Isoform, GA110, of Human Follicular Fluid Is Degraded by the Bovine Oviductal Fluid Component (소의 수란관액에 의한 사람 난포액의 Gelatinase A 동위효소인 GA110의 분해)

  • Kim, Min-Jung;Kim, Ji-Young;Leec, Seung-Jae;Yoon, Yong-Dal;Cho, Dong-Jae;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.5 no.1
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    • pp.23-33
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    • 2001
  • When mammalian oocytes ovulate into the oviduct, associating follicular fluid components are exposed to the oviductal environment, possibly resulting in the mutual interaction between fillicu1ar and oviductal fluids. In the Present study, we have demonstrated for the first time that components of fallicular fluid could be modified by the oviductal fluid. Gelatin zymographic analyses of human follicular fluid (hFF) obtained from IVF patients showed consistently the presence of 110 kDa gelatinase (GA110) in addition to many bands among which 62 kDa gelatinase was predominant. Addition of EDTA or phenanfhroline to the gelatinase substrate buffer during gel incubation abolished GA110 band whereas phenylmethylsulffnyl fluoride (PMSF) did not. In contrast, bovine oviductal fluid(bOF) exhibited only 62 kDa gelatinase. Surprisingly, when bOF was added to hFF in 1:1 ratio and then the mixture was incubated for 3 h at 37$^{\circ}$C, GA110 of hFF disappeared. Disappearance of GA110 by bOF was observed even within 30 min after mixing with hFF. Addition of aminophenylmercuric acetate (APMA) to hFF also abolished enzymatic activity of GA110 but increased the activityof 62 kDa gelatinase. However, APMA abolished many other gelatinases as well unlike bOF. Interestingly, treatment of hFF with EDTA for 3 h remarkably increased the enzymatic activity of GA110 but not that of other gelatinases. Addition of phenanthroline, PMSF or soybean trypsin inhibitor (SBTI) did not affect overall gelatinase activities. Again, addition of bOF to the hFF pretreated with any of the above proteinase inhibitors abolished the appearance of GA110. Human serum also showed GAI 10 of which activity was greatlyenhanced by EDTA treatment. Similar to hFF, serum GA110 also disappeared by the addition of bOF. Human granulosa cell homogenate did not reveal any appreciable gelatinase activity except 92 kDa gelatinase. Anti-human gelatinase A antibody reacted with 62 kDa gelatinase of hFF. Based upon these results, it is concluded that bOF could selectively degrade an isoform of gelatinase A present in hFF and human serum.

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The Role of Neutrophils and Epidermal Growth Factor Receptors in Lipopolysaccharide-Induced Mucus Hypersecretion (리포다당질 (lipopolysaccharide)에 의한 기관지 점액 생성 기전에서 호중구와 상피세포 성장인자 수용체 (epidermal growth factor receptor)의 역할)

  • Bak, Sang Myeon;Park, Soo Yeon;Hur, Gyu Young;Lee, Seung Heon;Kim, Je Hyeong;Lee, Sang Yeub;Shin, Chol;Shim, Jae Jeong;In, Kwang Ho;Kang, Kyung Ho;Yoo, Se Hwa
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.1
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    • pp.80-90
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    • 2003
  • Background : Goblet cell hyperplasia is a critical pathological feature in hypersecretory diseases of the airways. A bacterial infection of the lung is also known to induce inflammatory responses, which can lead to the overproduction of mucus. Recently, mucin synthesis in the airways has been reported to be regulated by neutrophilic inflammation-induced epidermal growth factor receptor (EGFR) expression and activation. In addition, it was reported that migration of the activated neutrophils is dependent on the matrix metalloproteinases (MMPs), especially MMP-9. In this study, bacterial lipopolysaccharide (LPS)-induced goblet cell hyperplasia and mucus hypersecretion by EGFR cascade, resulting from the MMPs-dependent neutrophilic inflammation were investigated in the rat airways. Methods : Pathogen-free Sprague-Dawley rats were studied in vivo. Various concentrations of LPS were instilled into the trachea in $300{\mu}{\ell}$ PBS (LPS group). Sterile PBS ($300{\mu}{\ell}$) was instilled into the trachea of the control animals (control group). The airways were examined on different days after instilling LPS. For an examination of the relationship between the LPS-induced goblet cell hyperplasia and MMPs, the animals were pretreated 3 days prior to the LPS instillation and daily thereafter with the matrix metalloproteinase inhibitor (MMPI; 20 mg/Kg/day of CMT-3; Collagenex Pharmaceuticals, USA). The neutrophilic infiltration was quantified as a number in five high power fields (HPF). The alcian blue/periodic acid-Schiff (AB/PAS) stain were performed for the mucus glycoconjugates and the immunohistochemical stains were performed for MUC5AC, EGFR and MMP-9. Their expressions were quantified by an image analysis program and were expressed by the percentage of the total bronchial epithelial area. Results : The instillation of LPS induced AB/PAS and MUC5AC staining in the airway epithelium in a time- and dose-dependent manner. Treatment with the MMPI prevented the LPS-induced goblet cell hyperplasia significantly. The instillation of LPS into the trachea induced also EGFR expression in the airway epithelium. The control airway epithelium contained few leukocytes, but the intratracheal instillation of LPS resulted in a neutrophilic recruitment. A pretreatment with MMPI prevented neutrophilic recruitment, EGFR expression, and goblet cell hyperplasia in the LPS-instilled airway epithelium. Conclusion : Matrix metalloproteinase is involved in LPS-induced mucus hypersecretion, resulting from a neutrophilic inflammation and EGFR cascade. These results suggest a potential therapeutic role of MMPI in the treatment of mucus hypersecretion that were associated with a bacterial infection of the airways.

Effect of PLA2 Inhibitor Rutin on Endotoxin-Induced Acute Lung Injury (내독소로 유도된 급성폐손상에서 PLA2의 억제제인 Rutin의 효과)

  • Kim, Seong-Eun;Lee, Young-Man;Park, Won-Hark
    • Applied Microscopy
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    • v.34 no.1
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    • pp.31-42
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    • 2004
  • Acute respiratory distress syndrome (ARDS) is a kind of acute lung injury characterized by inflammatory disruption of alveolar-capillary barrier and notorious for its high mortality. Neutrophils cause cell damage through the production of free radicals, inflammatory mediators, and proteases in ARDS. $PLA_2$ might serve a primary regulatory role in the activation of neutrophils. This present study was performed to elucidate the effect of rutin known as $PLA_2$ inhibitor on ARDS induced by endotoxin. Endotoxin had increased lung myeloperoxidase (MPO) activity, BAL (bronchoalveolar lavage) protein content, numbers of neutrophils in BALF (bronchoalveolar lavage fluid) compared with those of control rat (p<0.001). In addition, histological evidence of lung injury was correlated with neutrophil influx into alveolar space and cerrous perhydroxide granules were found in lining of endothelial cell, alveolar type I, II cells. In contrast, pretreated group of rutin had significantly decreased all of the parameters (p<0.001). These data suggest that inhibition of $PLA_2$ is one step approach that block the process of ARDS. Accordingly, we conclude that rutin can be used as the prophylactic agent for ARDS on the bases of these experimental results.

Effect of Paroxetine and Sertraline Treatment on Forced Swim Test-Induced Behavioral and Immune Changes in the Mouse (마우스 강제수영에 의한 행동 및 면역반응 변화에 대한 Paroxetine과 Sertraline의 효과)

  • Eum, Se-Yeun;Jeong, Min-Ho;Lim, Young-Jin;Kim, Bu-Kyung;Jeong, Soo-Jin;Hahn, Hong-Moo;Choe, Byeong-Moo
    • Korean Journal of Psychosomatic Medicine
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    • v.8 no.1
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    • pp.46-57
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    • 2000
  • Objectives : The purpose of the present study was to examine the effect of subacute treatment with the selective serotonin reuptake inhibitors(paroxetine and sertraline) on immobility in the forced swim test(FST) and on FST-induced changes in immune parameters of the mice. Methods : Authors applied a modified method of FST by Porsolt et al. Over 5 BALB/c mice were used for each group of experiments. To explore the changes in immune parameters by FST, authors investigated the production of anti-rat RBC antibody, concanavalin A(ConA)- or lipopolysaccharide(LPS)-stimulated splenocytes proliferation assay and cytokine gene expression. Results : Both paroxetine and sertraline decreased the duration of immobility in a dose-related manner. FST-performed mice showed a significant decrease in mitogenic responses of splenocytes and a slight increasing tendency in anti-rat RBC antibody response. All these responses were attenuated significantly by paroxetine and attenuated nearly nominal significance level by sertraline. The cytokine profiles of ConA-stimulated splenocytes from FST-performed mice showed stronger expression of IL-4 and weaker expression of IL-2 than control mice, and no changes in the expressions of IFN-$\gamma$ and lymphotoxin. IL-6 and IL-10 were not expressed in both group of mice. The pretreatment of paroxetine and sertraline attenuated the altered cytokine expressions in FST-performed mice to some extent. Some alterations of the expressions of IL-6 and IL-10 were observed in the mice which the selective serotonin reuptake inhibitors had been pretreated. Conclusion : The subacute treatment of paroxetine and sertraline attenuated the FST-induced behavioral and immune changes, and these serotonin reuptake inhibitors may exert some modulating effects on the immune system by the induction of cytokine gene expression, especially IL-6 and IL-10.

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Studies on the Site of Cardioaccelerating Action of Nicotine, DMPP, McN-A-343, AHR-602, Tyramine, Angiotensin and Neostigmine in Rabbits (가토(家兎)에서 Nicotine, DMPP, McN-A-343, AHR-602, Tyramine, Angiotensin, Neostigmine이 심박증가(心博增加)를 일으키는 작용점(作用點)에 관한 연구(硏究))

  • Bark, Pyong-Soon
    • The Korean Journal of Pharmacology
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    • v.11 no.2
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    • pp.29-40
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    • 1975
  • 1. Sites of the cardioaccelerating action of nicotine, DMPP, McN-A-343, AHR-602, tyramine, angiotensin and neostigmine were investigated in spinal rabbits. 2. The cardioaccelerating action of the above substances was substantially weak in reserpine-pretreated rabbits. The accelerating action was scarcely observed after propranolol administration. 3. Tetrodotoxin and guanethidine did not affect the cardioacceleration due to nicotine, DMPP, tyramine and isoproterenol, but they markedly weakened that due to McN-A-343, AHR-602, angiotensin and neostigmine. 4. Chlorisondamine blocked the cardioacceleration by nicotine and DMPP; atropine that by McN-A-343 and AHR-602. 5. Appropriate doses of isoproterenol, nicotine, DMPP, McN-A-343, tyramine, angiotensin and neostigmine, when administered into the right auricle, produced almost the same degree of cadia acceleration as when they were given to the right ear vein. AHR-602 did not produce significant cardioacceleration through this route. 6. Nicotine, DMPP and neostigmine when injected into the right auricle produced marked cardioacceleration, whereas they produced little action when injected into the left ventricle. Isoproterenol and tyramine produced more marked effect by the intraauricular route than the intraventricular one. 7. McN-A-343, AHR-602 and angiotensin produced more marked cardioacceleration by the intraventricular administration than the intraauricular one. The intraventricular AHR-602 produced marked cardioacceleration. 8. It is inferred that the sites of cardioaccelerating action of nicotine, DMPP, and tyramine will be either the terminals of the adrenergic nerves or the extraneuronal stores of norepinephrine and that of McN-A-343, AHR-602, angiotensin and neostigmine will be the adrenergic neurons in the heart. The sites on which nicotine, DMPP, tyramine and neostigmine will act are chiefly distributed in the auricular tissues and those on which McN-A-343, AHR-602, and angiotensin act chiefly in the ventricular tissues.

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Changes in Quality of Lettuce During Storage by Immersion-Type Hydrocooling (침지식 냉수냉각에 의한 상치의 저장중 품질변화)

  • Jeong, Jin-Woong;Kim, Byeong-Sam;Kim, Oni-Woung;Nahmgung, Bae;Park, Kee-Jai
    • Korean Journal of Food Science and Technology
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    • v.27 no.4
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    • pp.537-545
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    • 1995
  • Quality changes of lettuce were studied during storage to investigate the efficiency, cooling properties and the washing and storage effects of immersion-type hydrocooling. As a result of plotting the nondimensionalized lettuce temperature versus cooling time, its cooling rate coefficient was shown to be $-0.365\;min^{-1}{\sim}-0.255\;min^{-1}\;(R^2=0.99{\sim}0.88)$. Rate of weight loss was not significantly (p>0.05) different between lettuces with various treatment conditions during storage at $5^{\circ}C$. However, during storage at $15^{\circ}C$, weight loss of hydrocooled lettuce was lower than that of non-treated lettuce after from 10 to 15 days. Especially, The lettuce packed with PE was more effective than that packed with try in terms of packing condition after hydrocooling. Lettuce pretreated with sterilizing agent, packed with PE vinyl film, removed residual water after hydrocooling had lower decaying rate than any other lettuces. Changes of L and b values in hydrocooled lettuce were slower than those of non-treated one. During changes of chlorophyll content, the initial value, $115.7{\sim}147.3\;mg%$ was decreased to $50{\sim}60%$ after 25 days of storage at $5^{\circ}C$ and within 15 days of storage at $15^{\circ}C$. It could be presumed that the addition of sterilizing agent reduced the initial level of overall total and coliform count and its growth rate during storage. The respiration rate of hydrocooled lettuce at $5^{\circ}C$ was $23.95\;mg{\cdot}CO_2/kg\;hr$, which is 10% of those of non-treated lettuce.

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Characteristics of Hot-Water Extracts from Salmon Frame as Basic Ingredients for Gomtang-like Products (열수추출 연어 Frame 엑스분의 곰탕 유사 제품 베이스로서의 특성)

  • Han, Byung-Wook;Kim, Hye-Suk;Jee, Seung-Joon;Lee, Jae-Hyoung;Kim, Hyung-Jun;Park, Shin-Ho;Ji, Seong-Gil;Heu, Min-Soo;Kim, Jin-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1326-1333
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    • 2007
  • For preparing the basic ingredients of Gomtang-like products from the extracts of salmon frame, the extraction conditions of salmon frame were examined, and the characteristics of the extracts were compared with commercial Gomtang. Based on the crude protein, Ex-N and sensory attributes, the extractions were optimized by extracting pretreated-salmon frame in 12 times (v/w) of water for 12 hrs, before filtering with cheese cloth to yield 3 times the volume of the raw material. The concentrations of heavy metals in extracts from salmon frame were below the safety limits suggested by KFDA. The mai or amino acids were glutamic acid and aspartic acid as the free amino acids, and glycine, proline, and glutamic acid as the total amino acids. The calcium and phosphorus contents were 18.0 mg/100 mL and 33.1 mg/100 mL, respectively, and they accounted for 20% and 18% of the recommended daily allowance for mineral intake. The angiotensin I converting enzyme (ACE) inhibitory activity was improved by incubation with Flavourzyme for 4 hrs and its $IC_{50}$ was 2 mg/mL. The results above suggested that the enzymatic hydrolysates from extracts of salmon frame could be used as a basic ingredient for preparing Gomtang-like products.

The Role and Localization of Nitric Oxide Synthase in Neurogenic Inflammation of the Rat Airways (백서의 기도 선경성 염증에서 산화질소 합성효소(Nitric Oxide Synthase)의 역할과 분포)

  • Shim, Jae-Jeong;Lee, Sang-Yub;Lee, Sang-Hwa;Suh, Jung-Kyung;Kim, Chul-Hwan;Cho, Jae-Youn;In, Kwang-Ho;Yoo, Seo-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.3
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    • pp.420-433
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    • 1996
  • Background : There have been many debates about the effects of nitric oxide on the neurogenic inflammation. The role of nitric oxide in the neurogenic inflammation of airways will be required a better understanding of the localization and types of nitirc oxide synthase(NOS) activity in the neurogenic inflammation of airways. Method : To investigate the role of nitric oxide in airway neurogenic inflammation, 1) the effects of neurokinin receptor antagonist (FK224) and nitric oxide synthase inhibitor, $N^{\omega}$-nitro-L-arginine (L-NNA) on plasma extravastion were evaluated in four groups of Sprague-Dawley rats ; sham operation group(sham NANC group), electrical vagal stimulation group(NANC2 group), intravenous pretreatment groups with FK224 (1mg/kg ; FK224 group), and L-NNA(1mg/kg ; L-NNA group) 15 minutes before vagal NANC stimulation. 2) NOS activity in trachea with neurogenic inflammation was localized by immunohistochemical stain. Immunohistochemical stain was performed by antibodies specific for inflammatory cells(iNOS), brain(bNOS), and endothelium (eNOS) on trachea obtained from sham NANC, NANC2, and FK224 groups. Results : The results are that plasma extravsation in neurogenic inflammation of rat airways was inhibited by FK224, but enhanced by L-NNA pretreatment(P<0.05). There was significantly increased infiltration of inflammatory cells in subepithelium of neurogenic inflammatory trachea, but the reduction of subepithelial infiltration of inflammatory cells was observed after pretreatment with FK224(P<0.05). Immunostaining with anti-iNOS antibody showed strong reactivity only in infiltrated inflammatory cells in neurogenic rat trachea, and these iNOS reactivity was reduced by pretreatment with FK224. bNOS immunoreactivity was significantly increased only in the nerves both of neurogenic inflammatory and FK224 pretreated trachea compared with sham NANC trachea(p<0.05). eNOS immunoreactivity was not significant change in endothelium in neurogenic inflammation of rat trachea. Conclusion : These results suggest that nitric oxide released from iNOS in infiltrated inflammatory cells has main role in neurogenic inflammation of rat trachea. The presence of bNOS immunoreactivity in the nerves indicates that nitric oxide may be released from the nerves in rat trachea with neurogenic inflammation.

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EFFECTS OF SUBINHIBITORY CONCENTRATIONS OF ANTIMICROBIAL AGENTS ON CELL SURFACE PROPERTIES AND VIRULENCE FACTORS OF MUTANS STREPTOCOCCI (아저해농도(亞沮害濃度)의 항균물질이 mutans streptococci의 세포표면성질과 독력인자에 미치는 영향)

  • Kim, Young-Jae;Hahn, Se-Hyun;Lee, Sang-Hoon;Jang, Ki-Taeg;Kim, Chol-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.4
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    • pp.605-616
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    • 2004
  • Subinhibitory concentrations (sub-MICs) refer to concentrations below minimum inhibitory concentrations (MICs). The antimicrobial agents may be present at relatively high concentration, at least higher than bacterial MIC and thereafter be deserted off a surface and function at sub-MICs, perhaps by interfering with bacterial metabolism. Consequently, the aim of this study was to determine the effects of growth, in the presence of sub-MICs of antimicrobial agents, on the cell surface properties and virulence factors of mutans streptococci and to investigate the efficacy of a chemical approach in vitro. Streptococcus mutans Ingbritt and Streptococcus sobrinus 6715-7 were used. Eight antimicrobial agents (Sanguinaria extract;SG, Chlorhexidine digluconate;CHX, Fluoride;F, Propolis;PP, Hydrogen peroxide;HP, Triclosan;TC, Sodium dodecyl sulfate;SDS Cetylpyridinium chloride; CC) were diluted serially in broth to determine MICs and to compare the growth rate, acid production, hydrophobicity, adhesion activity to saliva coated hydroxyapatite, glucan synthesis and cellular aggregation of experiment groups (in the presence of sub-MICs) with those of control (in the absence of antimicrobial agents). Sub-MICs of antimicrobial agents affected the growth of cells, hydrophobicity, and adhesion of bacteria to saliva coated hydroxyapatite and glucan synthesis. They also resulted in a significant reduction in pH after 12 hours (p<0.05). By cells pretreated with proteinase K, either the aggregation induced by antimicrobial agents was completely inhibited or the aggregation titers were markedly increased. According to the results of the present study, each antimicrobial agent at sub-MICs could affect similar as its known action mechanism and could continually inhibit cariogenic bacteria at such concentrations. Thus, the use of these antimicrobial agents would be one of the effective methods to prevent dental caries.

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A Mineralogical Study on the Arsenic Behavior in the Tailings of Nakdong Mine (낙동광산의 광미 내 비소 거동에 대한 광물학적 연구)

  • Lee, Woo-Chun;Cho, Hyen-Goo;Kim, Young-Ho;Kim, Soon-Oh
    • Journal of the Mineralogical Society of Korea
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    • v.22 no.4
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    • pp.359-370
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    • 2009
  • Arsenic and heavy metals leached out as a result of oxidation of tailings exposed to the surface pose a serious environmental contamination of mine areas. This study investigated how arsenic behavior is controlled by a variety of processes, such as oxidation of sulfides and formation or alteration of secondary minerals, based on mineralogical methods. The study was carried out using the tailing samples obtained from Nakdong mine located in Jeongseongun, Gangwondo. After separating magnetic and non-magnetic minerals using pretreated tailing samples, each mineral sample was classified according to their colors and metallic lusters observed by the stereoscopic microscope. Subsequently, the mineralogical properties were determined using various instrumental analyses, such as x-ray diffractometer (XRD), energy dispersive spectroscopy (EDS), and electron probe micro analyzer (EPMA). The literature review confirmed that various ore minerals were identified in the Nakdong ore deposits. In this study, however, there were observed a few original ore minerals as well as secondary and/or tertiary minerals newly formed as a result of weathering including oxidation. In particular, we did not recognize pyrrhotite which has been known to originally exist in a large abundance, but peculiarly colloform-type iron (oxy)hydroxides were identified, which indicates most of pyrrhotite has been altered by rapid weathering due to its large reactivity. In addition, a secondary scorodites filling the fissure of weathered primary arsenopyrites were identified, and it is speculated that arsenic is immobilized through such a alteration reaction. Also, we observed tertiary iron (oxy)hydroxides were formed as a result of re-alteration of secondary jarosites, and it suggests that the environment of tailing has been changed to high pH from low pH condition which was initiated and developed by oxidation reactions of diverse primary ore minerals. The environmental change is mainly attributed to interactions between secondary minerals and parental rocks around the mine. As a result, not only was the stability of secondary minerals declined, but tertiary minerals were newly formed. As such a process goes through, arsenic which was immobilized is likely to re-dissolve and disperse into surrounding environments.