겔형태 인공누액이 각막실질세포 및 결막상피세포에 미치는 영향

이종수,김은희,김수진,이지은,김나미,Jong Soo Lee,Eun Hee Kim,Soo Jin Kim,Ji Eun Lee,Na Mi Kim 대한안과학회 2010 대한안과학회지 Vol.51 No.12

Purpose: To evaluate the biological effects and cytotoxicity of gel-type artificial tears on human corneal keratocytes and conjunctival cells in vitro. Methods: Human corneal keratocytes and conjunctival epithelial cells were exposed to Soothe® and Systane® at variable concentrations. Evaluations were conducted through an MTT-based calorimetric assay to measure the metabolic activity and through a lactate dehydrogenase (LDH) assay to assess cellular damage. Apoptotic response was examined using fluorescent microscopy and flow cytometric analysis, and cellular morphologic results were evaluated with a transmission electron microscope. Results: The inhibitory effects of corneal keratocyte and conjunctival cell proliferations increased at higher concentrations and longer exposure times to Soothe® and Systane®. The LDH titers increased after Soothe® exposure, but showed no significant difference after Systane® exposure. Soothe® and Systane® treatments both produced fluorescence, representing apoptotic cells. In flow cytometry, the maximal apoptotic response was observed for both types of artificial tears, although Systane® showed less edema, as well as reduced cytoplasmic and nuclear cell degeneration compared to those of Soothe®. Conclusions: The apoptotic responses of Soothe® and Systane® are associated with inhibitory effects of human corneal keratocyte and conjunctival epithelial cell proliferations. To inhibit the cellular proliferation of human corneal keratocytes and conjunctival epithelial cells, Systane® may be less severe than Soothe® at higher concentrations and longer exposure times. J Korean Ophthalmol Soc 2010;51(12):1643-1651

익상편조직의 섬유모세포에 미치는 사이클로스포린의 영향

이종수,이승욱,이상준,김나미,Jong Soo Lee,Seung Wook Lee,Sang Jun Lee,Na Mi Kim 대한안과학회 2012 대한안과학회지 Vol.53 No.3

Purpose: To evaluate the response and cellular damage of cultured human pterygial cells according to the concentration and exposure time of topical cyclosporin. Methods: Human pterygial cells were exposed to a cyclosporin A concentrations of 0.1 μg/ml (0.0001%), 1 μg/ml (0.0001%), 10 μg/ml (0.001%), 100 μg/ml (0.01%), or 500 μg/ml (0.05%) for 5 or 10 minutes. An MTT-based colorimetric assay was performed to assess the metabolic activity of cellular proliferation, and a lactate dehydrogenase (LDH) leakage assay was used to determine cellular damage. The extra-cellular matrix of PIP, laminin and MMP were evaluated, and the measurement of pro-inflammatory cytokine, TNF-α and IL-1b. IL-6, IL-8 was performed using ELISA kits. Results: The pterygial cellular inhibitory effect of cyclosporin was similar to that of the control according to the concentration and exposure time (p > 0.05). Compared with the control, the level of LDH did not show a statistically significant difference between concentration and exposure time (p > 0.05). There was no significant difference of inhibitory effects by PIP, laminin, or MMP between the experimental and control groups (p > 0.05). The production of TNF-α and IL from the experimental pterygial cells due to the effect of cyclosporin was not significantly different from that of the control at a longer exposure time or stronger concentration (p > 0.05). Conclusions: The response of pterygial cells to topical cyclosporin A at concentrations less than 0.05% for less than 10 minutes of exposure time showed no prevention of pterygial recurrence. With regard to cellular damage, little effects on inhibition of PIP, laminin, MMP, IL, and TNF-α were observed compared with that of the control. J Korean Ophthalmol Soc 2012;53(3):466-472

정상 및 원추각막에서 각막 전, 후면의 융기 및 시상면만곡도의 비교분석

이종수,이승욱,이승민,이미현,Jong Soo Lee,Seung Wook Lee,Seung Min Lee,Mi Hyun Lee 대한안과학회 2012 대한안과학회지 Vol.53 No.10

Purpose: To compare changes of anterior and posterior corneal elevation, and sagittal curvature according to the severity of keratoconus and to compare differences between keratoconus and normal cornea. Methods: A total of 81 eyes diagnosed with keratoconus and 20 eyes of normal subjects were evaluated with a Pentacam Scheimpflug camera. The keratoconus eyes were divided into 3 groups according to mean keratometer (K): mild (K ≤ 47.0 diopters (D)), moderate (47.0 to 52.0 D), and severe (52.0 D≥). The following parameters were obtained to evaluate the correlation of keratoconus: corneal thickness, anterior and posterior corneal elevation, and sagittal curvature. Results: Out of 81 keratoconus eyes, 56 eyes were mild, 12 eyes were moderate, and 13 eyes were severe keratoconus. The mean central corneal keratometer, anterior and posterior corneal elevation, and sagittal curvature of the keratoconus eyes were 49.7 D, 22.07 μm, 38.16 μm, 52.76 D and the values increased statistically compared to the normal eyes. Furthermore, the values increased significantly with the severity of keratoconus. ROC curve analysis showed the estimated meaningful value for anterior and posterior corneal elevation and sagittal curvature of keratoconus for diagnosis; there were no diagnostic values for corneal thickness and refractive power. Conclusions: The index of 5.5 μm for mean anterior elevation, 12.5 μm for mean posterior elevation, and 44.5 μm for mean sagittal curvature using the PentacamⓇ are useful to diagnose keratoconus. Variation of anterior and posterior elevation, and sagittal curvature measured by PentacamⓇ are useful in understanding the process of keratoconus. J Korean Ophthalmol Soc 2012;53(10):1432-1437

몇종의 세균과 Saccharomyces cerevisiae에 대한 식품첨가물의 향균 특성

이종수,오준세,김나미,금종화,이석건,Lee, Jong-Soo,Oh, Jun-Sei,Kim, Na-Mi,Keum, Jong-Hwa,Lee, Suk-Kun 배재대학교 자연과학연구소 1996 自然科學論文集 Vol.8 No.2

현재 식품첨가물로 많이 사용되고 있는 유기산과 안정제 및 색소 등의 각종 세균과 Sacch. cerevisiae 에 대한 향균 특성을 조사하였다. 산도 조절용으로 사용되고 있는 젖산, 사과산, 호박산 및 주석산은 L. acidophilus와 Sacch. cerevisiae에 대하여 향균성이 없었으나 B. subtilis등의 세균에 대하여는 향균성이 있었고 특히 사과산은 P. aeruginosa에 대하여 강한 향균력이 있었다.(최소생육저지농도 : 0.05%). 안정제로서의 알긴산과 펙틴은 B. subtilis, E. coli, P. aeruginosa에 대하여 비교적 강한 향균성을 보였고 L. acidophilus에 대하여는 향균성이 없었다. 황색 색소(홍화엘로우)와 적색 색소(Red powder-N)는 향균성이 없었고 표백제인 $NaHSO_3$의 세균에 대한 최소생육저지농도는 0.05%, Sacch. cerevisiae에 대하여는 0.5%로 향균성이 있었다. In order to survey the safety of some food additives, antimicrobial activity of acidulants, stabilizers, antioxidants, natural coloring materials and bleaching agents against 5 strains of bacteria and Sacch. cerevisiae were investigated by dilution method and minimal inhibitory concentration(MIC) method. Malic acid as acidulants displayed the effective antimicrobial activity in vitro against P. aeruginosa and its MIC is 0.05%. Alginic acid and pectin as stabilizer also displayed strong antimicrobial activity against B. subtilis, E. coli and P. aeruginosa, and tannin(antioxidants) and $NaHSO_3$ displayed antimicrobial activity against all bacteria tested. However gums(Arabia, Xanthan, Gua) and natural coloring materials(Hongwha Yellow, Red powder-N) were not affected to growth of bacteria and Sacch. cerevisiae.

Neisseria lactamica 2118이 생성하는 $\beta$-Galactosidase의 정제 및 성질

이종수,곽인영,김나미,Lee, Jong-Soo,Kwak, In-Young,Kim, Na-Mi 배재대학교 자연과학연구소 1991 自然科學論文集 Vol.4 No.-

p-Aminopheny1-$\beta$-D-thiogalactopyranoside agarose 친화성 크로마토그라피를 이용하여 Neisseria lactamica 2118이 생성하는 $\beta$-galactosidase를 정제한 후 몇가지 효소학적 성질을 조사 하였다. Neisseria lactamica 2118이 생성하는 $\beta$-galactosidase는 구성효소로서 lactose와 IPTG에 의하여 유도되지 않았다. 정제효소의 작용최적온도는 $35^{\circ}C$, pH는 7.5이었고 $50^{\circ}C$로 15~60분 처리시 약 80%의 활성이 유지되었으며 pH 6.0~9.0에서 안정 하였다. 또한 $Hg^(2+)$와 $Co^(2+)$에 의하여 그 활성이 저해 되었다. $\beta$-Galactosidase(EC 3.2.1.23) from pathogenic Neisseria lactamica 2118 was purified by p-aminopheny1-$\beta$-D-thiogalactopyranoside agarose affinity chromatography. Then some properties of the purified $\beta$-galactosidase were investigated.$\beta$-Galactosidase form Neisseria lactamica 2118 was constitutive enzyme, not induced by lactose and IPTG. Optimal activity was observed at $35^{\circ}C$ and pH 7.5 and the enzyme was stable at the range of pH 6.0-9.0 and at temperature below $50^{\circ}C$. The enzyme activity was inhibited by cations such as $Hg^(2+)$ and <Tex>$Co^(2+)$.

화침을 이용한 흉요추부 압박골절 치험 1례

이종수,한순현,Lee, Jong-Soo,Han, Soon-Hyun 척추신경추나의학회 2002 대한추나의학회지 Vol.3 No.1

Stable compression fractures are most often caused by trauma as traffic accidents. Stable compression fractures can refer pain to the mid-back, which is characterized by the pain in the thoracolumbar spinous prosess. recently we experienced a 19 years old man who had lumbago originating from Stable compression fractures and condition was improved through burning acupunture. 흉요추 이행부의 안정성 압박골절은 통증의 소실과 후만각 증가 억제에 의한 척추의 안정화이다. 이의 원인을 후방인대의 손상에 의한 손상으로 보고 프로로테라피(Prolotherapy)로 이의 치료를 시행 할 수 있다고 생각되며 한방적인 방법으로 화침을 적용할 수 있었다. 최근 저자는 흉요추 이행부의 안정성 압박골절에 의한 요배통 환자를 발견하고 화침치료를 통해 증세 회복에 도움이 되었기에 치료내용과 경과를 문헌고찰과 함께 보고하는 바이다.

잉어 내장중의 Mouse 치사성분 Cyprinol Sulfate의 분리 및 생리 활성

이종수,Lee, Jong-Soo 생화학분자생물학회 1994 한국생화학회지 Vol.27 No.5

양식산 잉어(Cyprinidae, Cyprinus carpio) 내장의 MeOH 추출물로부터 mouse의 치사성분을 분리 정제하였다. 이 성분은 HRFABMS로부터 분자식은 $C_{27}H_{48}O_{8}S$로 판명되었으며 NMR 등 각종 기기 분석 자료를 검토한 결과, 잉어과 어류의 담낭 중에 존재한다고 알려진 독특한 성분인 cyprinol sulfate($5{\alpha}-cholestane$ $3{\alpha}$, $7{\alpha}$, $12{\alpha}$, 26, 27-pentol, 26-sulfate)로 동정되었다. Cyprinol sulfate의 mouse치사량은 2.0~3.0mg/MU 이었고, 약한 용혈성($500{\mu}g/ml$)과 prolyl endopeptidase 활성 저해작용($IC_{50}$ > $200{\mu}g/ml$)을 나타내었으나 Bacillus subtilis와 Aspergillus niger에 대한 항균성이나 L1210 세포에 대한 독성은 나타내지 않았다. A toxic component to mice was isolated from the viscera of cultivated carp (Cyprinus carpio) as white powder. Molecular formula was obtained to be $C_{27}H_{48}O_{8}S$ on the basis of the HRFABMS data. NMR spectra of the isolated compound were corresponded with those of $5{\alpha}-cyprinol$ except the $C_{26}$ sulfated carbon. Therefore, it was identified as cyprinol sulfate ($5{\alpha}-cholestane$ $3{\alpha}$, $7{\alpha}$, $12{\alpha}$,26,27-pentol 26-sulfate). It showed toxicity to mice (2~3 mg/MU, i.p.), hemolytic activity ($500{\mu}g/ml$) and prolyl endopeptidase inhibition ($IC_{50}$ > $200{\mu}g/ml$), but, had no antibacterial, antifungal and cytotoxic activity to L1210 cells.

β-galactosidase를 생성하는 고온성(高溫性) 세균(細菌)의 생리적(生理的) 특성(特性)에 관한 연구(硏究)

이종수,김찬조,Lee, Jong Soo,Kim, Chan Jo 충남대학교 농업과학연구소 1982 Korean Journal of Agricultural Science Vol.9 No.1

고온균(高溫菌)의 생리적(生理的) 특성(特性)과 내열기구(耐熱機構)에 관한 기초자료(基礎資料)를 얻고자 온천토양(溫泉土壤)에서 ${\beta}$-galactosidase를 생산(生産)하는 고온균(高溫菌)을 분리(分離)하여 동정(同定)하고, 몇가지 주요(主要)한 생리적(生理的) 특성(特性)을 검토(檢討)하였으며, 그의 균체지방산(菌體脂肪酸) 조성(組成)을 gas chromatography로 분석(分析)하여 다음과 같은 결과(結果)를 얻었다. 1. ${\beta}$-galactosidase를 생산(生産)하는 공시(供試) 균주(菌株)는 Thermus속(屬)으로 동정(同定)되었다. 2. 분리선정(分離選定)한 균주(菌株)의 최적(最適) 생육온도(生育溫度)는 $65^{\circ}C$이었고, $37^{\circ}C$에서 생육(生育)하지 않는 절대(絶對) 고온균(高溫菌)이었으며 최적(最適) pH는 6.5내외(內外)이었다. 3. NaCl에 대(對)한 내성(耐性)은 NaCl 1% 이상(以上)에서 생육(生育)하지 못했다. 4. 공시균주(供試菌株)의 항생물질(抗生物質) 내성(耐性)은 Penicillin G는 $10{\mu}g/m{\ell}$, Chloramphenicol은 $0.5{\mu}g/m{\ell}$이었다. 5. Vitamin 요구성(要求性)은 Ca-pantothenate와 pyridoxine-HCl을 절대적(絶對的) 생육인자(生育因子)로 요구(要求)하였고 niacin을 자극적(刺戟的) 생육인자(生育因子)로 요구(要求)하였다. 6. 공시균주(供試菌株)는 최대(最大) 흡수파장(吸收波長)이 420nm 부근(附近)의 색소(色素)를 생성(生成)하였고 그의 생성능(生成能)은 약(弱)하였다. 7. 공시균주(供試菌株)의 균체 지방산(脂肪酸) 조성(組成)은 palmitic acid 60.20%, lauric acid 11.80%, myristic acid 7.56%, behenic acid 4.25%, capric acid 1.77%, stearic acid 2.13%, arachidic acid 1.53%이었다. This experiment was carried out to elucidate the thermotolerant properties of a thermophilic bacterium which was isolated from soils of the hot springs area and selected for the ${\beta}$-galactosidase production. This bacterium was identified as a strain belong to the genus Thermus. Biochemical and physiological characteristics of this strain were studied, including the investigation of the fatty acid composition of its neutral fats. The results obtained were summarized as follows. 1. Optimal temperature and pH for growth of this strain were $65^{\circ}C$ and pH 6.5 respectively, and it was found to be an absolute thermophilic bacterium which could not grow at the temperature below $43^{\circ}C$. 2. No growth was obtained in the medium which contained more than 1.0% of sodium chloride. 3. The tolerable concentration of antibiotics were 10mg of penicillin G per ml of medium and 0.5mg of chloramphenicol per ml respectively. 4. This strain had auxotrophilic requirements for calcium-pantothenate and pyridoxin-HCl as an essential factor and for niacin as a stimulative factor. 5. Yellow pigment was released into the liquid culture of this strain, which showed maximum absorption at 420 nm. 6. Fatty acid composition of neutral fats of the strain was palmitic acid, 60.20%; lauric acid, 11.80%; myristic acid, 7.56%; behenic acid, 4.25%; capric acid, 1.77%; stearic acid, 2.13%; arachidic acid, 1.53%; and others unidentified, 10.7%.

검정해면으로부터 항균성을 가진 방선균의 분리 동정 및 항균물질의 구조

이종수,최종덕,LEE Jong-Soo,CHOI Jong-Duck 한국수산과학회 1998 한국수산과학회지 Vol.31 No.4

검정해면 (Halichondlia okadai)에 공생하는 것으로 추정되는 항균성을 가진 방선균주를 분리하여 SEM, 형태학적, 화학 분류학적 방법에 의하여 균주를 동정하였으며, 이 균주를 marin broth 2216 배지에서 대량 b배양하여 균사체로부터 항균물질을 각종 HPLC에 의하여 분리하고 X선 결정 구조 해석. MS, NMR spectrum에 의하여 구조를 확인하였다. ISP 2 배지에서 $30^{\circ}C$, 7일간 배양 결과, 기질균사는 갈색을 띄었고 그 위로 ivory색의 기중균사를 형성하였다. spore chain은 flexibilis로서 10개 이상의 oval 형의 포자가 사슬을 형성하고 있었으며 표면은 smooth하였다. 한편. 화학적 특성으로는 L-form의 DAP를 가지며 특징적인 당이 없는 cell wall type I 이었고. 인지질은 PII형 이었으며, 주요 지방산은 iso-14 : 0, anteiso-15 : 0, iso-16 : 0, 16 : 0 및 iso-17:0 이었다. 이들 결과와 Bergey's Manual에 의해 이 균주는 Strptomyces sp.로 동정하였다. 대량 배양한 동결 건조 균체 70g으로 부터 10mg의 연한 노란색의 항균 물질을 분리 정제하였다. 질량 분석에서 이 물질의 분자량은 466으로 확인되었고, 이를 benzene-acetone 용매중에서 결정화하여 X선 결정 구조 해석을 행한 결과 indolo[2,3-a]carbazole alkaloid의 staurosporine 으로 판명되었으며 proton 및 carbon NMR data도 이와 잘 일치하였다. An Actinomycetes strain, MSA-1, containing antimicrobial component was isolated from the black sponge, Halichondzia okadai, and was identified to a genus level by morphological and chemotaxonornic methods. The gray colored spores were oval type with smooth surface and formed flexibilis spore chains. The cell wall of this strain was type I containing D-aminopimellic acid (D-DAP) and no specific sugar was detected. Phospholipid of the cell membrane was PII type including phophoethanolamine and the major fatty acids of total lipid were branched anteiso-15 : 0, iso-16 : 0, 16 : 0 and iso-17 ; 0. From these results and other characteristics described in the Bergey's Manual, this strain was identificated as a Streptomyces sp. Meanwhile, 10mg of pale yellow colored antimicreobial component was isolated by HPLC method from the cultured Streptomyces sp. (70g of cryophillized mycellis). By crystallographyc analysis, HIRESMS and NMR assignment, the antimicrobial component produced from the strain MSA-1 was elucidated as the staurosporine (indolo[2,3-a]carbazole alkaloid).

양식산 굴로부터 쓴맛 성분의 분리 및 성질

이종수,LEE Jong-Soo 한국수산과학회 1995 한국수산과학회지 Vol.28 No.1

1989년 11월부터 1990년 1월 사이에 전남 가막만에서 채취한 양식 굴 및 이를 원료로 제조된 훈제 기름담금 통조림, 보일드 통조림에서 강한 쓴맛이 나타나 품질상 문제가 발생하였는바, 본 연구에서는 이러한 쓴맛의 원인 물질을 규명하고저 시도하였다. 이들 시료의 아세톤 추출액을 액-액 분배, 알루미나, 규산 및 ODS 등의 각종 칼럼을 이용하여 정제한 결과, 5개의 쓴맛성분을 분리하였으며, 이들은 질량분석(FAB-MS)으로 부터 분자량이 각각, OY-22 : 837, OY-23 : 851, OY-24 : 821, OY-25, OY-26 : 835 이었으며, proton NMR spectrum 및 각종 기기분석 결과, 6-7개의 아미노산으로 이루어진 환상 peptide로 추정되었다. 또한, 아미노산 분석으로부터 OY-24에서는 Val, Leu 이, OY-25에서는 Leu, Ile이, OY-26에서는 Leu, Leu이 각각 확인되었으며, 나머지는 이상 아미노산으로 생각되었다. 이들 성분들은 마우스에 대하여 급성독성은 없었으며 $(100{\mu}g/20g\;mice,\;I.p.)$, Asp. niger E. coli, Bac. subtilis 에 대하여는 항균성도 없었다$(10{\mu}g/disk).$ 현재, 각종 생리활성 및 정확한 구조를 검토 중에 있다. Five bitter taste compounds (OY-22, OY-23, OY-24, OY-25 and OY-26) were firstly isolated from cultured oyster (Crassostrea gigas) at Gamak Bay, Sourthern coast of Korea, between November, 1989 and January, 1990, and smoked-canned oyster, which were produced by the same oysters. They were presumed as cyclic peptides composed with 6 or 7 amino acids, including sulfur on the basis of NMR and MS spectra. Val and Leu in OY-24, leu and lie in OY-25 and tow leucines in OY-26 were detected from those each compounds, seperately, by the amino acid analysis. Another amino acids were thought as non-constitutional amino acids. They showed non-toxic to mice $(100{\mu}g/20g\;mice i.p.)$ and non actibacterial artivities to Asp. niger and B. subtilis $(10{\mu}g/disk)$. The chemical structures and other biological activities of them are now in studying.