• Journal of Korean Ophthalmic Optics Society
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• v.12 no.3
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• pp.19-25
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• 2007

It was studied how using soft contact lens multi-purpose solution (MPS), often used for medical treatment, effects the inhibition on cell growth, and how using the MPS demages eye cells, on rabbit eye's corneal epithelium and endothelium tissue. $ReNu^{(R)}$ (Baush & Lomb, USA), Opti-free $express^{(R)}$ (Alcon, USA), Free-sol $plus^{(R)}$ (Hanamedicon, Korea) were used as MPS. After culturing Clone 1-5C-4 cell lines (Human conjunctival cell lines), cell growth inhibition rate was measured by MIT assay. By making Hematoxylin and Eosin stain specimen, the morphology was observed by optical microscope. In the In vivo experiment, 9 white rabbit eyes (18 eyes) were classified into 3 groups. The experiment group is left eyes (9 eyes) of rabbit, and MPS were dropped; however the control group, the right eyes (9 eyes), were only used a saline solution without a preservatives. After the dropping within the period, the cornea surface of rabbit endothelium tissue was observed using scanning electron microscopy (SEM).

• Journal of Korean Ophthalmic Optics Society
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• v.18 no.4
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• pp.525-531
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• 2013

Purpose: To identify the apoptosis caused by Proparacaine hydrochloride (PPC), a topical anesthesia, applied to conjunctival cell lines and determine whether pigallocatechin-gallate (EGCG), has protective effects on. Methods: The conjunctival cell lines were treated with 0.5% of Alcaine$^{(R)}$, 0.5% of PPC and 0.01% of Benzalkonium chloride (BAC) for 15 minutes, respectively in order to investigate the effects of topical anesthesia on cells, and followed by cultured for 12 and 24 hours. The recovery effects were investigated by measuring level of cellular proliferation inhibiting using MTT assay and LDH assay. The conjunctival cell lines were pre-treated with EGCG $10{\mu}M$ for 3 hrs and post-treated with 0.5% PPC for 15 mins in order to investigate whether EGCG has protective effects, flow cytometry were performed in order to observe apoptosis. Results: A result of the additional culture of 12 and 24 hours and again immediately after the treatment for 15 minutes 0.5% of Alcaine$^{(R)}$, 0.5% of PPC, the 0.01% of BAC, cell viability was not increased in all groups (p<0.05). The cell viabilities were higher than in cells 3 hours post-treated with $10{\mu}M$ of EGCG and pre-treated PPC 0.5% (68.2%), compared to cells ($32.2{\pm}2.0%$) treated only with 0.5% of PPC. PPC 0.5% also induced apoptosis in the treated group was reduced by the addition of EGCG. Conclusions: It is considered that the EGCG has cell protective effects when it is added to PPC, a topical anesthesia, by improving cell viability and inhibiting apoptosis.

• Journal of Korean Ophthalmic Optics Society
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• v.13 no.3
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• pp.95-101
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• 2008

Purpose: Hydrogen peroxide which is one of the reactive oxygen species has been seen to cause various diseases, various cellular disinfections, gene transformation and cell death. The goals of this study were to determine the protective effect of EGCG against $H_2O_2$-induced apoptotic death in conjunctival cell lines. Methods: We measured cell viability by MTT assay and analyzed DNA fragmentation to check up a distinctive feature in cell death and measured the removal ability of free radicals by DPPH free radical scavenging assay and evaluated the oxygen free radical's quantity in the cell by DCFH-DA assay. The mRNA expression in the cell were examined by RT-PCR. Results: Cell viability and free radical scavening activites was significantly increased in dose dependently after cell was exposed to EGCG. And DNA fragmentation and intracellular ROS was decreased. It was showed the mRNA expression which increase of bcl-2, bcl-xL expression and decrease of bax expression. Conclusions: From these results, it suggests that EGCG has an antioxidant effect and protects conjunctival cell lines from the $H_2O_2$-mediated apoptosis through the modulation of the mRNA expression.

• Journal of Korean Ophthalmic Optics Society
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• v.9 no.2
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• pp.333-343
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• 2004

Impression cytology refers to application of cellulose acetate filter material to the ocular surface to remove the superficial layers of the conjunctival epithelium. The technique is non-invasive, is easy to perform, causes minimal discomfort to the patient, and can be used to follow changes in the conjunctival ocular surface over time. With this method, the morphology of the conjunctival ocular surface can be studied and the degree of squmaous metaplasia assessed. This study was performed to evaluate the conjunctival surface by impression cytology in dry eye patients. A total of 70 students with no contact lens wearing history were recruited. Subjects were required to fill in a McMonnies dry eye symptom questionnaire. The non-invasive tear thinning time(TIT) test of each subject was measured, followed by Schirmer tear test(STI), tear film break-up time(TBUT) tests and Rose-bengal staining were performed as a baseline. Conjunctival epithelial cells from the inferior bulbar conjunctiva were harvested by the impression cytology technique. The specimens collected were labelled and stained with PAS(Periodic Acid Schift)-haematoxylin. The goblet cells and conjunctival epithelial cells were observed under a light microscope of 400x magnification. The specimens were classified according to the Nelson Grading scale which was based on the degree of squamous metaplasia such as changes of goblet cells density, size/form, N:C(nucleus : cytoplasm) ratio. Dry eye patients were observed morphological changes of the epithelial cells, different nuclear alterations, decrease of the goblet cells density. The degree of cytological changes was related to severity of dry eye conditions. When the epithelial cell morphology was graded according to the system described by Nelson, specimens from the control group revealed 91.43% of the eyes to be grade 0 and 8.57% to be grade 1, whereas of the dry eye patients, 20% were grade 0, 42.86% grade 1, 34.29% grade 2 and 2,86% grade 3. Impression cytology represents a non- or minimally invasive biopsy of the ocular surface epithelium with no side effects or contraindications. It has demonstrated to be a useful diagnostic aid for a wide variety of processes involving the ocular surface. This technique is a safe, simple method and may help increase understanding of various ocular surface alterations in dry eye patients.

• Journal of Korean Ophthalmic Optics Society
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• v.9 no.1
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• pp.173-180
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• 2004

This study aims to investigate the effects of hydrogen peroxide and grapefruit seed extract used as a chemical and natural disinfectants on human conjunctival cells in vitro. The main component of grapefruit seed extract is a narigin. It is one of the flavonoid types in citrus fruits and f1avonoids are widely recognized as naturally occurring(삭제) antioxidants. Cytotoxicity was determined by mitochondrial activity(MTT assay) and DNA damage was analyzed by measuring Comet assay. In LDH assay, 5% of grapefruits seed extract has been observed as a material is giving recovery effect of damaged cultured conjuctival cells by hydrogen peroxide. And also, each of concentrations has been treated simultaneously with same amounts and cytotoxicity of hydrogen peroxide and grapefruit seed extract have been estimated by LDH leakage assay after 24 hours. In conclusion, H2O2-induced cytotoxicity, apoptosis were Significantly prevented by grapefruit seed extract. It is a main component of bioflavonoids that we can simply take it as food. The present results suggest that grapefruit seed extract is a useful disinfectanct having antioxidant and antiapoptopic activity as a natural product.

• The Journal of Korean Society of Virology
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• v.26 no.1
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• pp.23-30
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• 1996

대략 36,000 base pairs (bp)의 두 가닥짜리 DNA를 지놈으로 가진 사람 아데노바이러스 (Ad)는 DNA 상동성(相同性) 및 생물학적/생화학적 성격이 특이한 49개의 혈청형이 알려져 있는데, 이들 대부분의 Ad가 영유아군 및 면역능이 저하된 성인에서 치사적 결과를 초래할 수 있다. Ad의 세포향성(向性)(tropism)은 매우 다양하여 종류에 따라 상기도 감염, 각결막염, 영유아 장염등을 유발하는데 최근 Ad의 다양한 병원성에 대한 원인을 분자생물학적 수준에서 규명하려는 노력의 일환으로 지역에 따라 주되게 출현하는 Ad형 규명이 활발히 이루어지고 있다. Ad 동정/확인은 표면을 이루고 있는 group 공통항원인 hexon 단백질을 탐지하는 효소면역 측정법 (EIA)에 의하며, Ad형별은 Ad fiber의 세포독성 중화시험에의 한다. 그러나, 세포독성 중화시험이 엄청난 노동력 및 시간을 요구하면서도 민감도/특이도가 만족스럽지 못하여 이를 개선하기 위하여 검체 또는 세포배양에서 Ad DNA를 추출하여 제한효소 절단형태를 비교하는 방법이 개발되었는데 이는 세포배양에 잘 자라지 않는 바이러스주의 형별뿐만 아니라 지역 분리주들의 지놈 변형주를 관찰하는 분자생물학적/분자역학적 연구에도 도움이 되고 있다. 국내에는 Ad와 관련된 소아장염의 빈도가 rotavirus에 의한 것 다음으로 빈번한데도 Ad40/41외에 주되게 출현하는 장내 Ad형들이 전혀 규명된 바 없고, 한국형 Ad들의 지놈형태가 전혀 보고된 바가 없다. 또한 세계적으로 Ad형별 조사지역이 늘어감에 따라 유아장염과 연관된 Ad 역시 Ad40, 41이 외의 형들이 Ad40, 41을 능가하는 것으로 보고되고 있는 지역도 있으나 국내에서는 Ad40, 41이외의 형들은 그 역학적 중요도가 전혀 알려져 있지 않다. 이로서 본 연구의 목적은 Ad주들에 특이 중화항체를 이용한 세포독성 중화시험과 Ad DNA 절단법을 적용하여 한국형 장내 Ad주들의 형별을 처음으로 시도함과 동시에 1989-1991사이 출현한 Ad들의 유전적 변형을 관찰하려는 것이었다. 두 방법 모두 사용하였을 때 주되게 출현하는 장내 Ad형들은 Ad4l, Ad2, Ad7, Ad5, 및 Ad40이었다. Ad40/41-양성 검체를 제외한 Ad hexon-EIA양성들의 77.5%를 형별 할 수 있었던 Ad DNA의 제한효소 절단방법은 형들간의 교차중화로 특이성이 낮았던 중화방법 (47.5%)보다 매우 효율적이어서 두 가지 방법을 함께 적응하였을 때는 40주중의 81.5%인 35주를 형별 할 수 있었다. 또한Ad DNA 제한 효소 절단방법은 Ad7 변이주 (Ad7b)도 탐지 할 수 있었다.

• Journal of Korean Ophthalmic Optics Society
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• v.10 no.4
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• pp.293-304
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• 2005

As the eye irritant test of lens washing agent, ReNu$^{TM}$ was analysed using Draize methods (1959) according to KFDA Guidelines. In addition, to test the potential toxicity of test articles the ratio of inflammatory cells and non-inflammatory epitheloid cells were also observed using smear cytology methods against ocular discharge. At test, the histopathological changes on the cornea, iris, retina and sclera were also observed in all animals. Slight irritancy of the cornea and conjunctiva were observed at 1, 2 and 3 days after dropping in non-washing group. The Mean Index of Ocular Irritation(MIOI) of these points are detected as 4.17, 3.00 and 1.33, respectively. In washing group, slight irritancy of the cornea and conjunctiva were observed at 1 and 2 days after dropping with MIOI as 0.67 and 1.33, respectively. Therefore, ReNu$^{TM}$ was considered as non-irritating materials because the MIOI is detected below 5.00 throughout the whole experimental periods in both washing and non-washing groups. The Index of Acute Ocular Irritation(IAOI) is also detected as 4.17(1 day after dropping). Except of the somewhat increase trend of the inflammatory cell ratios in ocular discharge at 1 day after dropping of non-washing group, but significances are not detected, on the other hand, no meaningful changes on smear cytology of ocular discharges are observed in this study. In addition, no abnormal histopathological changes on the cornea, iris, retina and sclera were also not detected in ReNu$^{TM}$ dropping group compared to that of non-treated control eyes.

• Journal of Veterinary Clinics
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• v.27 no.6
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• pp.751-754
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• 2010

A 6-year-old spayed female Persian cat presented with a 3-month history of recurrent ulcerative keratitis with noticeable opacification and vascularization of the right cornea. The lesion was nonresponsive to topical antibiotics and to nonsteroidal anti-inflammatory drugs. Ophthalmic examination showed signs of ocular discomfort, such as epiphora and blepharospasm, in the right eye. Biomicroscopic examination revealed an irregular, edematous, vascularized mass with pink to white tissue on the entire cornea and mild conjunctivitis. A tentative diagnosis of feline proliferative eosinophilic keratitis (FPEK) was made on the basis of clinical appearance. Cytologic examination of the cornea showed a mixture of numerous eosinophils and mast cells, which confirmed the original diagnosis of FPEK. The cat was treated with a topical antibiotic-corticosteroid combination, cyclosporine ointment, trifluridine eye drops, and oral Llysine. The clinical signs improved remarkably 18 days after the cat was first examined. The short-term use of corticosteroids and long-term use of cyclosporine and an anti-viral agent resolved the lesion without recurrence of the disease for 1 year.

• Journal of Veterinary Clinics
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• v.30 no.3
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• pp.210-213
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• 2013

We herein describe a feline case of facial dermatitis whose histopathological features resembled to those of FHV-associated ulcerative dermatitis. A 3-year-old, intact male domestic short-haired cat was presented with 2-years history of pruritic dermatitis that initially appeared on periocular area and extended toward the entire face. The cat had ocular discharge and conjunctivitis from 2-month of age. Clinically, skin lesions were characterized as erythema, erosions and ulcers covered with crusts on the facial and perianal area. Histopathologically, the facial lesion was characterized as interface dermatitis with hydropic degeneration at the basal layer, and single cell necrosis of keratinocytes. In addition, the epidermal and dermal necrosis infiltrated with eosinophils, and intranuclear inclusion bodies in keratinocytes were also recognized. Moreover, feline herpesvirus-1 gene was detected by a PCR analysis using a swab obtained from the crusted lesions. Based upon these findings, the present case was considered as having FHV-associated ulcerative dermatitis. Therapy including oral acyclovir and topical recombinant feline interferon omega resulted in marked improvement of the skin and mucosal lesions.

• Journal of Veterinary Clinics
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• v.21 no.3
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• pp.229-235
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• 2004

To characterize the recent outbreaks of canine distemper (CD) in Korea, we carried out epidemiological investigations by clinical observations, serum neutralizing (SN) antibody titer determination and RT-PCR on the 315 dogs which were clinically suspected as canine distemper virus (CDV) infection. One hundred and sixty two of 315 dogs were infected with CDV. Breed or gender did not seem to have effects on the prevalence of CD. The major part of dogs were in young age from 6 weeks to 18 weeks of age, and were not vaccinated or incompletely vaccinated. Clinical signs of dogs with CD were multi systemic and extremely variable. Dogs died from CD had significantly more ocular signs and neurologic signs than those of dogs survived (p<0.05). The SN titers against CDV of 157 (96.9%) dogs were under 1:16, which is less than protective level. One possible explanation for recent outbreaks of CD in Korea might be low antibody titers against CDV because of vaccination failure. Therefore, to reduce the impact of virulent infection in the dog population, dogs should be vaccinated adequately and prophylactic measures should include isolation of young dogs from the dog population until vaccination can be expected to provide protection.