• Title/Summary/Keyword: Epithelial cells

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Time Course Change of Phagocytes and Proinflammatory Activities in BALF in Endotoxin-induced Acute Lung Injury (시간별 내독소 정맥주입으로 유발된 급성폐손상의 변화양상에 대한 고찰)

  • Moon, Seung-Hyug;Oh, Je-Ho;Park, Sung-Woo;NamGung, Eun-Kyung;Ki, Shin-Young;Im, Gun-Il;Jung, Sung-Whan;Kim, Hyeon-Tae;Uh, Soo-Tack;Kim, Yong-Hoon;Park, Choon-Sik;Jin, Byeng-Weon
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.2
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    • pp.360-378
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    • 1997
  • Background : Severe acute lung injury(ALI), also known as the adult respiratory distress syndrome(ARDS), is a heterogenous nature of dynamic and explosive clinical synrome that exacts a mortality of approximately 50%. Endotoxin(ETX) is an abundant component of the outer membrane of gram-negative bacteria capable of inducing severe lung injury in gram-negative sepsis and gram-negative bacterial pneumonia, which are among the most common predisposing causes of ARDS. The influx of PMNs into airway tissue is a pathological hallmark of LPS-induced lung injury. And there is a substantial evidence suggesting that cytokines are important mediators of lung injury in gram-negative sepsis. However, the kinetics of phagocytes and cytokines by an exact time sequence and their respective pathogenic importance remain to be elucidated. This study was performed to investigate the role of phagocytes and proinflammatory cytokines in ETX-induced ALI through a time course of changes in the concentration of protein, $TNF{\alpha}$ and IL-6, and counts of total and its differential cells in BALF. The consecutive histologic findings were also evaluated. Method : The experimental animals, healthy male Sprague-Dawley, weighted $200{\pm}50g$, were divided into control- and ALI- group. ALI was induced by an intravenous administration of ETX, 5mg/kg. Above mentioned all parameters were examined at 0(control), 3, 6, 24, 72 h after administration of ETX. $TNF{\alpha}$ and IL-6 cone. in BALF were measured by a bioassay. Results : The protein concentration and total leukocyte count(TC) in BALF was significantly increased at 3h compared to controls(p < 0.05). The protein conc. was significantly elavated during observation period, but TC was significantly decreased at 72h(p < 0.05 vs. 24h). There was a close relationship between TC and protein cone. in BALF(r = 0.65, p < 0.001). The PMN and monocyte count was well correlated with TC in BALF, and the correlation of PMN(r = 0.97, p < 0.001) appeared to be more meaningful than that of monocyte(r = 0.61, p < 0.001). There was also a significant correlation between protein cone. and PMN or monocyte count in BALF(PMN vs. monocyte : r = 0.55, p < 0.005 vs. r = 0.64, p < 0.001). The count of monocyte was significantly elavated during observation period though a meaningful reduction of PMN count in BALF at 72h, this observation suggested that monocyte may, at least, partipate in the process of lung injury steadly. In this study, there was no relationship between IL-6 and $TNF{\alpha}$ cone., and $TNF{\alpha}$ but not IL-6 was correlated with TC(r = 0.61, p < 0.05) and monocyte(r = 0.67, p < 0.05) in BALF only at 3, 6h after ETX introduced. In particular, the IL-6 cone. increased earlier and rapidly peaked than $TNF{\alpha}$ cone. in BALF. In histologic findings, the cell counts of lung slices were increased from 3 to 72h(p < 0.001 vs. NC). Alveolar wall-thickness was increased from 6 to 24h(p < 0.001 vs. NC). There was a significant correlation between the cell counts of lung slices and alveolar wall-thickness(r= 0.61, p < 0.001). This result suggested that the cellular infiltrations might be followed by the alterations of interstitium, and the edematous change of alveolar wall might be most rapidly recovered to its normal condition in the process of repair. Conclusion : We concluded that although the role of PMN is partly certain in ETX-induced ALI, it is somewhat inadequate to its known major impact on ALL Alveolar macrophage and/or non-immune cells such as pulmonary endothelial or epithelial cells, may be more importantly contributed to the initiation and perpetual progression of ETX-induced ALI. The IL-6 in ETX-induced ALI was independent to $TNF{\alpha}$, measured by a bioassay in BALF. The early rise in IL-6 in BALF implies multiple origins of the IL-6.

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Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.21-27
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    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).

Expression of Epidermal Growth Factor Receptor in the Inflamed Gingival Epithelium and the Dental Follicle (염증성 치은 상피와 치낭의 표피성장인자 수용체의 발현 및 실험적 치아이동에 미치는 영향에 관한 연구)

  • Kim, Young Ho;Bae, Chang
    • The korean journal of orthodontics
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    • v.27 no.2
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    • pp.349-357
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    • 1997
  • Epidermal growth factor(EGF), a single chain polypeptide of 53 amino acids with a molecular weight of 6,045 Da, was first isolated from the male mouse submandibular glands. EGF stimulates cellular proliferation and differentiation in several tissues and accelerates the rate of wound healing. EGF is bound to the specific receptor(EGFR) on the cell membrane of its target cell. EGFR is a transmembrane glycoprotein with a molecular weight of 170,000 Da and is detectable on a large variety of cell types and tissues. The authors investigated the expression of EGFR in the normal and inflamed human gingival epithelium to study the role of EGFR in the inflammation of the gingival epithelium, and the expression of EGFR in the dental follicle by using in situ mRNA hybridization and immunohistochenistry. The results weree as follows : 1. The expression of EGFR mRNA in the normal gingival epithelium on in situ mRNA hybridization was mainly localized on the basal cell layer, and the spinous layer was weakly positive The granular and cornified layers were negative 2. The expression of EGFR protein in the normal gingival epithelium on inmunohistochemistry was localized on the cornified and granular layers, and the spinous layer was weakly positive. The basal cell layer was completely negative 3. The expression of EGFR mRNA in the inflamed gingival epithelium on in situ mRNA hybridization was evenly and homogeneously distributed in the whole layers of the gingival epithelium except the cornified layer. The staining intensity appeared to increase progressively from the basal cell layer to the cornified layer. 4. The expression of EGFR protein in the inflamed gingival epithelium on immunohistochemistry was evenly and homogeneously distributed in the whole layers of the gingival epithelium. The staining intensity appeared to increase progressively from the cornified layer to the basal cell layer. 5. Strong positive reaction was seen in the epithelial cell rests of Malassez, whereas only background staining was seen in other cells of the dental follicle. In conclusion, the up-regulation of EGFR in the inflamed gingival epithelium and the high amounts of EGFR in the epthelial cell rests of Malassez in the dental follicle can be regarded as responses to the possible damages to the oral environment to maintain the homeostatic conditions.

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Effect of Resistant Starch on the Large Bowel Environment and Plasma Lipid in Rats with Loperamide-Induced Constipation (저항전분 투여가 Loperamide 유도 변비 쥐의 대장환경과 혈청지질에 미치는 효과)

  • Sin, Hyun-Ju;Kim, Kwang-Ok;Kim, Sung-Hong;Kim, Young-Ah;Lee, Hye-Sung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.5
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    • pp.684-691
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    • 2010
  • The present study was conducted to evaluate the effect of resistant starch (RS) on the large bowel function and plasma lipids in rats with constipation induced by Loperamide. Animals were divided into six groups: normal control-5% cellulose, constipation-5% cellulose, constipation-5% pectin, constipation-5% RS-type 2 (RS2), constipation-8% RS2 and constipation-5% RS type 3 (RS3) groups, and fed experimental diets for five weeks. The results from RS groups were compared with those from other dietary fiber groups. The groups supplemented with RS3 or high level of RS2 showed significantly increased counts of bifidobacteria in the cecum than the other groups. The production of total short chain fatty acids in the cecal contents was significantly high in pectin, RS3 and high RS2 groups. The pH in the cecal contents of the RS supplemented groups was significantly decreased compared with the cellulose supplemented groups. The production of prostaglandin E2 in the colon mucus of the RS groups was higher than the normal group; however, it was significantly decreased compared to the cellulose or pectin supplemented constipated groups. The thickness of the mucus layer and the production of mucus from epithelial cells were significantly increased in RS3 group compared to the constipated cellulose group. Supplementation of resistant starch significantly elevated the ratio of HDL-cholesterol to total cholesterol and significantly lowered plasma atherogenic index compared with cellulose or pectin supplementation in constipated rats. The results of the present study demonstrated that resistant starch supplementation may help in improving the large bowel environment by stimulation of bifidobacterial proliferation, reduction of pH and inflammation factor and by increased production of mucus. It has also been found that an additional health benefit is improvement in lipid levels of serum.

Transport Properties of Aromatic Amino Acids by Amino Acid Transporter TAT1 (아미노산 수송체 TAT1에 의한 방향족 아미노산의 수송특성)

  • 김윤배;김명수;윤정훈;박주철;국중기;정해만;최봉규;정규용;김종근
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.775-781
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    • 2002
  • The T-type amino acid transporter 1 (TATI) is a Na$^{+}$-independent amino acid transporter which selectively trans- ports aromatic amino acids subserving the amino acid transport system T. To understand the transport properties of aromatic amino acids by human TAT1 (hTATl ), we have examined the hTATl -mediated aromatic amino acid transports using a Xenopus laeuis oocyte expression system. When expressed in Xenopin laeuis oocytes, hTATl induced L- [$^{14}$ C]tryptophan transport which was not dependent on Na$^{+}$ or Cl$^{[-10]}$ in the medium. Uptake was time-dependent and exhibited a linear dependence on incubation time up to 30 min. The L- ($^{14}$ C)tryptophan uptake was highly inhibited by L-isomers of tryptophan, tyrosine and phenylalanine, whereas other L-amino acids did not inhibit hTATl -mediated L- ($^{14}$ C)tryptophan uptake. The hTATl induced the relatively low-affinity transport of aromatic amino acids such as L- ($^{14}$ C)tryptophan, L- ($^{14}$ C)tyrosine and L- ($^{14}$ C)phenylalanine (Km values: 450~750 $\mu$M), consistent with the properties of classical amino acid transport system T. The L- ($^{14}$ C)tryptophan uptake did not show any remarkable pH dependence within the pH range of 5.5 to 8.5. The time-dependent efflux of L- ($^{14}$ C)tryptophan was detected from the oocytes expressing hTATl, which was not affected by the presence or absence of L-tryptophan in the extracellular medium, indicating that hTATl-mediated transport is due to the facilitated diffusion. Expression of hTATl in Xenopu laevis oocytes induced the transport of tryptophan, tyrosine and phenylalanine, indicating that hTATl is a transporter subserving system T These results suggest that hTATl has essential roles in the absorption of aromatic amino acids from epithelial cells to the blood stream. Hecause hTATl is proposed to be crucial to the efficient absorption of aromatic amino acids from intestine and kidney, its defect such as blue diaper syndrome could be involved in the disruption of aromatic amino acid transport.ort.

Molecular Cloning of Novel Genes Specifically Expressed in Snailfish, Liparis tanakae (꼼치, Liparis tanakae에서 특이하게 발현되는 새로운 유전인자의 검색)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.4 no.1
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    • pp.67-77
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    • 2000
  • Snailfish usually lives at the bottom of the sea and showed typical retrogressive change with specialized tissue structures of skin and skeletons. In order to obtain the specific genes of snailfish, highly expressed in the body, we made subtracted cDNA library and analyzed 200 clones. Totally 200 clones were obtained and sequenced, and among them 62 clones were turned out to be homologous to the known gene, i.e., thioesterase (9), myosin (8), creatine kinase (7), skeletal alpha-actin (6), parvalbumin b (5), ribosomal protein (5), type I collagen (3), muscle troponin (3), dopamine receptor (2), histatin (2), and heat shock protein (2), cystatin (1), lectin (1), statherin (1), secretory carrier membrane protein (1), keratin type I (1), desmin (1), chloroplast (1), muscle tropomyosin (1), reticulum calcium ATPase (1), ribonucleoprotein (1). The remaining 138 clones were low homologous or non-redundant genes through Genbank search. Especially 5 clones were novel and specifically expressed in the body tissues of Snailfish by in situ hybridization. Therefore, we analysed these 5 clones to identify the C-terminal protein structures and motifs, and partly defined the roles of these proteins in comparison with the expression patterns by in situ hybridization. C9O-77, about 5000 bp, was supposed to be a matrix protein expressed strongly positive in epithelium, myxoid tissue, fibrous tissue and collagenous tissue. C9O-116, about 1500 bp, was supposed to be a transmembrane protein which was weakly expressed in the fibrous tissue, epithelium tissue, and myxoid tissue, but strong in muscle tissue. C9O-130, about 1200 bp, was supposed to be an intracytoplasmic molecule usually in the epithelial cells. C9O-161, about 2000 bp, was weakly expressed in epithelium, muscle tissue and myxoid tissue, but specially strong in epithelium. C9O-171, about 1000 bp, was supposed to be a transcription factor containing zinc finger like domain, which was intensely expressed in the epithelium, muscle tissue, fibrous tissue, and in collagenous tissue.

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Cryptocaryoniasis of cultured flounder, Paralichthys olivaceus in low temperatures (저수온 양식 넙치 Paralichthys olivaceus의 Cryptocaryoniasis)

  • Ji, Bo-Young;Kim, Ki-Hong;Park, Soo-Il
    • Journal of fish pathology
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    • v.10 no.2
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    • pp.97-111
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    • 1997
  • In the winter of 1995, mass mortality occurred in cultured flounder, Paralichthys olivaceus in Gurongpo, Kyoungbuk, Korea. From the observations of moribund and dead fish, parasitic ciliates, which were shown as white spots to the naked eye, were considered to be involved in the mass mortality. From heavily infected flounders, histopathological, morphological and biological characterization of these ciliates were carried out. In the histological observation, many ciliates were found under the epithelia of gill filaments and skin, and caused hyperplasia of epithelial and mucus cells at the infected areas. The ciliates found on the body surface, fins and gills were very similar to Cryptocaryon irritans. However the ciliates showed two different patterns of reproductian, i.e., typical form(palintomy)and atypical form(budding plus multiple fission) at $16^{\circ}C$ of water temperature. The occurrence ratio between typical and atypical form was about 3:2. Tomitogenesis takes 8-14 days in the typical and 13-15 days in the atypical form. In the viability test at different temperatures and salinities, the typical form died below 30‰ at $12^{\circ}C$, below 20‰ at $16^{\circ}C$, below 15‰ at $20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. On the other hand, the atypical form died below 20‰ at $12^{\circ}C$, below 15‰ at 16-$20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. The results suggested that the atypical has better viability at low salinity than that of the typical at low temperatures. In the excystment time and success rates of excystment according to temperatures, the typical form showed 8 days, 30% at $12^{\circ}C$ : 6.5 days, 50%, at $16^{\circ}C$ : 5.5 days, 75% at $20^{\circ}C$ : and 7 days, 10% at $24^{\circ}C$, respectively. On the other hand, the atypical form showed 15.5 days at $12^{\circ}C$ : 14 days, 76.6% at $16^{\circ}C$ : 12 days, 72.2% at $20^{\circ}C$ : 10 days 31.6% at $24^{\circ}C$, respectively. The results suggested that the atypical form had longer excystment time than that of the typical form at any temperature and showed better stability at low temperatures.

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Cytopathologic Diagnosis of Bile Obtained by Percutaneous Biliary Drainage (담즙의 세포병리학적 진단에 관한 연구)

  • Park, In-Ae;Ham, Eui-Keun
    • The Korean Journal of Cytopathology
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    • v.3 no.1
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    • pp.1-11
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    • 1992
  • From the one hundred forty eight patients with evidence of biliary tract obstruction, 275 bile samples were obtained from percutaneously placed biliary drainage catheters. Of the 148 patients, ova of Clonorchis sinensis were demonstrated in 17 patients (11.5%), with the epithelial cells. Among them, one case also demonstrated coexisting adenocarcinoma. In 105 patients, the medical records were available for review and the clinical diagnoses were malignancy in 99 patients and benign lesion in 6 patients. Of the 99 patients in which clinico-radiologic diagnosis were malignant, cytologic results were positive in 23.2%. Dividing the patients Into two groups, the ones with tumor of bile duct origin (group I) and the others with tumors producing extrinsic compression of bile duct, such as periampullary carcinoma, pancreas head carcinoma or metastatic carcinoma in lymph nodes from tumors of adjacent organs (group II), the cytologic results were positive in 37% and 11.6%, respectively. In patients with histologic confirmation, the positive correlation was found in 50% and 20% in group I and group II, respectively, with remarkable difference between two groups. There were no false positives in cytologic diangosis. The overall concordance rate of cytologic diagnosis with diagnosis of clinical investigation in both benign and malignant lesions was 27.6% and the diagnostic specificity was 100%.

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HISTOPATHOLOGICAL STUDIES ON THE DISEASE CAUSED BY PATHOGENIC BACTERIA OF THE CARP (병원성세균에 의한 잉어의 궤양성질병에 관한 조직병리학적 연구)

  • PARK Soo-Il;CHUN Seh-Kyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.7 no.1
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    • pp.15-27
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    • 1974
  • Since the summer of the year 1972 ulcer disease of common carp, colour carp ana goldfish had suddenly spread widely and caused a serious mortality at fish farms especially around Gim-hae and Yang-san, near Busan. The present study aimed to find out the causative organisms, histopathological changes and the way of treatment, and the results are summarized as follows : Two kinds of pathogenic bacteria, i. e, Chondrococcus columnaris and Aeromonas sp. were isolated from the mucus of the lesion. According to the macroscopic findings, these symptoms began with the hemorrhagic spots under scales which fell out, after the collapse of the dermis, which was followed by lesion to form ulcer, and then the muscle was exposed. The fin was eroded mostly from the distal part, but sometimes from the basal part. Gills showed grey colour, and this part of the tissue collapsed. According to the histopathological findings, the dermis was exposed after the collapsed of the epithelium of the skin, and the necrosis of the musculature occurred, the muscular fibre being destroyed. The epithelial cells of gill tissue proliferated, thus gill filaments were conglutinated and collapsed. Fatty degeneration happened at the liver but the other organs seem to be normal. Treatment with the following mixtures were effective the water temperature of $22\~25^{\circ}C$, but not effective when the temperature was under as low as $13\~15^{\circ}C$. Mixture 1. Aivet (HB-115.HCI) 0.3ppm Malachite green 0.2ppm. Dipterex 0.4ppm Mixture 2. Furanace (P-7138) 0.05ppm Malachite green 0.2ppm Dipterex 0.4ppm When lesion healed, the epithelium, dermis, and the muscular fibre were regenerated.

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The Effect of Brown Tinted or UV-A blocking Ophthalmic Lens Against the Photooxidation of A2E, a Lipofuscin in Retina (망막 내 노인성 형광색소의 광산화에 미치는 Brown 착색렌즈와 UV-A 차단 안경렌즈의 영향)

  • Park, Sang-Il
    • Journal of Korean Ophthalmic Optics Society
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    • v.17 no.1
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    • pp.91-97
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    • 2012
  • Purpose: This purpose of study is to investigate the effect of UV-A-blocking or brown-tinted ophthalmic lens against A2E photooxidation which known as one of the etiologies of AMD(Age-related macular degenaration). Methods: The photooxidation of A2E, synthetic product of two molecules of all-trans-retinal and ethanolamine, was induced by the exposure to blue light (420~470 nm, $94mW/cm^2$) for 3 minutes. The inhibitory effect of UVblocking or brown-tinted ophthalmic lens against A2E photooxidation was evaluated by UV absorbance and HPLC analysis of remained A2E after the exposure to blue light. Results: UV-blocking ophthalmic lens could not inhibit A2E photooxidation induced by blue light irradiation. There was no difference in A2E photooxidation in the presence of brown-tinted ophthalmic lens to block 15% of visible ray, however, those lenses blocking 55% or 86% of visible ray showed the inhibitory effect of A2E photooxidation as 9.98% and 16.55%, respectively. By HPLC analysis, the amount of residual A2E which was not blocked by any lens was $199.29{\pm}26.53{\mu}M$, however, the inhibitory effect against A2E photooxidation was shown in the presence of brown-tinted lens. The remained A2Es were $264.58{\pm}31.91{\mu}M$ and $402.93{\pm}28.68{\mu}M$ in brown-tinted lenses of 55% and 86% blocking visible ray, respectively. However, there was no inhibitory effect against A2E photooxidation in the case of UV-blocking lens by HPLC analysis. Conclusions: In this study, brown-tinted ophthalmic lens was confirmed to have the inhibitory effect against the photooxidation of A2E, a causing substance of AMD onset.