• 미생물학회지
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• 제36권4호
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• pp.306-311
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• 2000

Herpes simplex virus type-1 (HSV-1)의 감염에 따른 세포내 유리 칼슘농도의 변화에 대한 실험을 수행한 결과, HSV-1이 Vero 세포에 감염한 후 4시간째에 세포내 칼슘농도가 최대로 감소한 것을 알았으며 이러한 세포내 유리 칼슘농도의 감소는 감염성 바이러스의 양에 따라 커지며, 유전자 발현 억제제의 처리나 바이러스의 불활성화에 의해 극복되었다. 따라서 바이러스의 유전자발현이 세포내 유리 칼슘농도의 감소에 중요한 역할을 한다는 것을 알 수 있다. 또한 Vero 세포에 바이러스를 감염시키고 미세소관 안정제인 taxol을 처리하여 4 시간째의 세포내 유리 칼슘농도의 감소가 극복된다는 사실로부터 바이러스이 유전자 물질의 이동에는 미세소관이 관여한다는 것을 알 수 있었다. 이와 같은 실험 결과로부터 Vero 세포에서 HSV-1에 의해 유도되는 세포내 유리칼슘 농도의 감소는 HSV-1 증식과 밀접한 관계를 가진다고 생각된다.

• Pediatric Infection and Vaccine
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• 제3권2호
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• pp.207-213
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• 1996

Herpes simplex virus(HSV) infections of the CNS are associated with significant morbidity and mortality even when appropriate antiviral therapy is administered. HSV infections of the brain can be subdivided into two categories : neonatal HSV infections, which usually are caused by HSV type 2, and herpes simplex encephalitis(HSE), which occur in patients over 3 months old and is nearly uniformly caused by HSV type 1. The clinical presentation of HSE is one of the focal encephalopathic process associated with altered levels of consciousness, fever, focal seizures and hemiparesis. But because of the lack of pathognomic clinical presentation and diagnostic procedure, the efforts to develop alternative diagnostic procedure have led to the use of new diagnostic technique such as polymerase chain reaction(PCR). We report a case of HSV type 1 encephalitis in 13 month old male infant who presented with altered level of consciousness, fever and focal seizures. With the use of the PCR, HSV-1 DNA was detected in cerebrospinal fluid from the patient. The symptoms and signs of encephalitis subsided by treatment with acyclovir in 14 days.

• 한국한의학연구원논문집
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• 제1권1호
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• pp.477-493
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• 1995

In order to facilitate the molecular characterization of the Herpes simplex Virus types 1 and types 2 genome DNAs, a gene library of cloned restriction frtgments have been produced. The Vero cells were infected with HSV-1 and HSV-2. 48 hours after infection, the infected cells Ivere Iysed, and multinucleated giant cells were observed approximately at seventy-two hours postinfection. The multiplication of HSV-1 and HSV-2 was observed in Vero cells using electromicroscopy. The nucleocapsids in nuclei were obseryed, and the assembled virions were budded out through the vacuole, and the virions were released from the cells. HSV-1 and HSV-2 was analyzed by digestion of their genome DANs with restriction ensymes. HSV-1 and HSV-2 genome DNAs were digested with BarnHI, Bgfl respectively. The BarnHI rlestriction fragments of HSV-1 and HSV-2 genome DNAs were twenty-seven fragments and thair molecular sizes were ranging $0.70{\sim}15.08$, $4.4{\sim}31.0$ tilobases. The BglII restriction fragments of HSV-1 and HSV-2 genome DNAs were sixteen, eighteen fragments and thair molecular sizes were ranging $4.8{\sim}30.0$, $1.2{\sim}25.0$ kilobases. And then BglII restriction frgments were cloned in Escherichia coli(E.coil) using the plasmid vector pBacPAK9.

• 한국한의학연구원논문집
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• 제2권1호
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• pp.417-429
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• 1996

Herpes simplex viruses(HSV) are one of the most common infectious virus of man. Though chemotherapies and antibiotics against HSV have been developed in many countries, but anti-HSV agents were not satisfactory to mankind by their toxic reaction and side effects. In order to search for anti-HSV agents from Korean traditional prescriptions, we extended the number of specimens. Both methanol extract and boilling water extract of the Korean traditional prescriptions were screened to detect anti-HSV activities by MTT assay. Korean traditional prescriptions showing anti-HSV activities as methanl extracts were Paekyopsan, Chesupwilyungtang, Yongdamsagantang, and prescription 11. Four methanol extracts showing anti-HSV activities were freationated by hexane and their efficacies were tested. Hexane freationations of Paekyopsan, Chesupwilyungtang, and prescription 11 showed in anti-HSV activities both haxane and methanol fractionation.

• 대한바이러스학회지
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• 제29권3호
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• pp.175-182
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• 1999

An attractive target for anti-herpes chemotherapy is the herpes simplex virus 1 (HSV-1) protease encoded by the UL26 gene. HSV-1 protease is essential for DNA packaging and virus maturation. To perform high throughput for potent inhibitors, the efficient production of larger amounts of highly purified enzyme and protease activity assay method must be established. In this report, expression in E. coli and purification of the protease gene of HSV-1 strain F was investigated. The protease gene was cloned pET28, and the nucleotide sequence of protease catalytic domain of HSV-1 compared strain F with other strains (KOS and CL101). In these results the F strain was different in base sequence. However, the amino acid sequence was identifical. The HSV-1 protease was purified with His-tagged affinity column. The analysis of HSV-1 protease activity was performed by high performance liquid chromatography.

• 대한약리학회지
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• 제22권2호
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• pp.123-127
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• 1986

4-carbon fatty acid인 n-Butyrate(n-BTA)는 herpes virus의 일종인 Epstein-Bat virus(EBV)에 작용해서 잠복형인 EBV를 활성형태로 유도시키는 것으로 알려져 있다. 고로 본 실험에서는 mouse의 삼차신경절에 잠복하고 있는 HSV의 재활성화에 대한 n-BTA의 효과를 실험 관찰하였다. Pentobarbital로 마취시킨 mouse의 양쪽눈 각막을 30 gauge 주사바늘로 scarify한 후에 type I HSV(HSV-1) $10{\mu}1(1{\times}10^5$ plaque-forming units)를 각각 점안 감염시켰다. virus를 감염시킨 4주 후에 mouse의 삼차신경절을 적출하여 시험관 내에서 조직배양을 시행하였다. 조직배양시에 0. 1, 0. 25, 0.5. 1.0 그리고 2.0mM농도의 n-BTA를 첨가하였으며 1일, 2일, 3일간 각각 배양한 후 신경절을 연마하여 연마액내의 HSV-1 titer를Vero cell monolayer에서 plaque assay로 측정하였다. 1) n-BTA첨가군은 잠재성 HSV가 대조군에 비하여 현저하게 빨리 재활성화 되었고 재활성화되는 virus의 양도 현저히 증가되었다. 2) 24시간을 계속해서 n-BTA 각 농도를 첨가해서 배양할 군은 n-BTA 6시간 첨가 배양하고 새로운 배양액으로 갈아서 18시간 배양한 군에 비해 잠재성 virus의 재활성화가 현저히 증가되었다. 3) Gang1ionic latent HSV-1의 재 활성화에 영향을 미치는 각 농도의 n-BTA는 Vero cell의 monolayer에서의 HSV-1의 번식에는 아무런 영향을 미치지 않았다.

• BMB Reports
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• 제36권4호
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• pp.379-386
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• 2003

A mutant herpes simplex virus 1, mtHSV, was constructed by inserting the E. coli beta-galactosidase gene into the loci of icp34.5, the apoptosis-inhibiting gene of HSV. The mtHSV replicated in and lysed U251 (human glioma cells), EJ (human bladder cells), and S-180 (mice sarcoma cells), but not Wish (human amnion cells) cells. With its intact tk (thymidine kinase) gene, mtHSV exhibited susceptibility to acyclovir (ACV), which provided an approach to control viral replication. An in vivo test with mtHSV was conducted in immune-competent mice bearing sarcoma S-180 tumors, which were treated with a single intratumoral injection of mtHSV or PBS. Tumor dimensions then were measured at serial time points, and the tumor volumes were calculated. Sarcoma growth was significantly inhibited with prolonged time and reduced tumor volume. There was microscopic evidence of necrosis of tumors in treated mice, whereas no damage was found in other organs. Immunohistochemical staining revealed that virus replication was exclusively confined to the treated tumor cells. HSV-1 DNA was detected in tumors, but not in the other organs by a polymerase chain reaction analysis. From these experiments, we concluded that mtHSV should be a safe and promising oncolytic agent for cancer treatment.

• 생약학회지
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• 제30권1호
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• pp.34-39
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• 1999

To search for less toxic antiherpetic agents, the inhibitory effects of twelve kinds of flavonoids including chrysin, quercetin, quercitrin, rutin, fisetin, gossypin, kaempferol, morin, naringenin, naringin, hesperetin and hesperidin on the plaque formation of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in Vero cells were examined by plaque reduction assay in vitro. Some flavonoids tested in this study showed potent antiherpetic activity, reducing intracellular replication of herpes simplex viruses when Vero cell monolayers were infected and subsequently cultured in medium containing flavonoids. Naringenin showed the most potent antiviral activity against HSV-1 with selectivity index (SI) of 19.1 and hesperetin showed the most potent antiviral activity against HSV-2 with SI of 9.8. These results suggest that some flavonoids may be a potential therapeutic agent for the treatment and prevention of herpes simplex virus infections.

• 약학회지
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• 제42권6호
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• pp.607-612
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• 1998

In the course of our search for anti-Herpes simplex virus type I (HSV 1) substances from natural sources, we screened crude drugs for their antiviral activity using SRB assay. T he methanol extract from herb of Brassica rapa (Cruciferae) was found to inhibit HSV-1. Though bioassay-directed fractionation of the extract, anti-HSV-1 agent was isolated by chromatographic separation using Amberlite XAD-4 and Sephadex LH-20. The structure of compound I was elucidated by spectral means including $^1H-^1H$ COSY, HMQC and HMBC to be isorhamnetin 3-O-${\beta}$-D-glucopyranoside (compound I). Compound I was active against HSV-1 with the 50% effective concentration of O.42mg/ml and the 50% cytotoxicity of 5.0mg/ml.

• 생약학회지
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• 제29권4호
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• pp.277-282
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• 1998

In order to search for anti-Herpes simplex virus type 1(HSV-1) agents, we screened 80 specimens of Korean traditional medicine by SRB assay. The methanol extracts of Thujae orientalis Semen (Cupressaceae) showed strong anti-HSV activity among samples tested. From the butanol fraction of Thujae orientalis Semen anti-HSV-1 agent was isolated by chromatographic separation using Amberlite XAD-4 and Sephadex LH-20. The structure was elucidated by spectroscopic methods, and was identified as ${\beta}-sitosterol$ (compound I). Compound I exhibited anti-HSV-1 activity with $EC_{50}$ of 0.6 mg/ml and $CC_{50}$ of 5.99 mg/ml, respectively.