• Journal of Pharmacopuncture
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• v.7 no.2
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• pp.29-41
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• 2004

Objective : This experimental studies were performed in order to prove the effect of Hirudin Herbal-acupuncture by using rats that had neuronal damage due to the Middle Cerebral Artery Occulsion(MCAO). Method : We observed the change of extracellular concentrations(${\mu}M$) of dopamine, DOPAC, HVA, HIAA, glutamate, aspartate, GABA, glysine, taurine, alanine, and tyrosine as extracted by vivo microdialysis, in the Hirudin Herbal-acupuncture administrated rats($240{\sim}260g$, Sprague-Dawley) subjected to the MCAO. The dialysates were extracted three times before the MCAO and six times after the MCAO every 20 minutes, and analysed by highperformance liquid chromatography(HPLC). Results : Hirudin Herbal-acupuncture significantly inhibited glutamate, aspartate, and tyrosine which are stimulant neurotransmitters at brain ischemia, and it significantly decreased glycine, GABA, taurine, and alanine which are inhibitory neurotransmitters at brain ischemia. Conclusion : Hirudin Herbal-acupuncture may prevent delayed neuronal death(DND) in selectively vulnerable focal areas of the brain effectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.5
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• pp.303-305
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• 2003

Scale-up of hirudin production from Saccharomyces cerevisiae from bench-scale to pilot-scale was carried out based on constant volumetric oxygen transfer coefficient (K$\sub$L/a). Fed-batch mode of cultivation using step-wise feeding strategy of galactose was employed for the production of hirudin in a 30-L and a 300-L pilot-scale fermentor. The final hirudin concentrations were achieved 390 mg/L and 286.1 mg/L, and the volumetric productivities were 80.4% and 90.7% with the 30-L and 300-L fermentors, respectively, compared to the productivity of the 5-L bench-scale fermentor.

• YAKHAK HOEJI
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• v.41 no.1
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• pp.74-80
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• 1997

A polygonal antibody against recombinant hirudin was raised for the development of a ELISA in biological fluids. Recombinant hirudin was conjugated to maleimide activated carrie r protein, KLH and injected to a rabbit. The third booster collection of antiserum was used as primary antibody for the ELISA. The titer for the detection antibody was determined. The direct ELISA could determine the concentration of hirudin in the range of ~10ng/ml. Affinity pulified IgG was obtained and conjugated to horseradish peroxidase. Purified IgG and IgG-HRP could be used as capture and detection antibody, respectively. Although sandwich ELISA would not give the satisfactory results. it could apply for the detection of hirudin level in the range of ~20 ${\mu}$g/ml.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.179-183
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• 1999

A recombinant form of hirudin, a potent thrombin-specific inhibitor derived from the bloodsucking leech, was expressed as a secretory product in Saccharomyces cerevisiae under the control of GALl0 promoter and the mating factor $\alpha$pre-pro leader sequence. In an attempt to produce recombinant hirudin (r-Hir) of therapeutic purity in large quantities, the fed-batch fermentation was carried out by using this recombinant yeast, and subsequently downstream processing was developed with the preparative-scale column chromatography systems. About 234 mg/l of biologically active r-Hir was produced as a secretory product by the fed-batch fermentation strategy developed for an efficient downstream processing. Using a two-step chromatography process (an anion exchange chromatography followed by the reverse phase HPLC), the r-Hir was purified to>98% with an overall recovery yield of 84%. According to the N-terminal amino acid sequencing, the purified r-Hir was found to have the predicted N-terminal amino acid sequence. The biological activity of the purified r-Hir to inhibit thrombin was also identical to that of the commercial hirudin.

• Korean Journal of Agricultural Science
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• v.24 no.1
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• pp.55-62
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• 1997

Hirudin is a potent thrombin inhibitor originally derived from the medicinal leech. Leeches were commonly used at the turn of the century for bloodletting and are still used today in some countries. So, hirudin is being developed as a alternative to heparin which has several shortcomings. This study is intended to analyze R&D trends, potential market for hirudin and existing anticoagulant drug market. Anticoagulant drug market in Korea is estimated by over 1,200 billion won in 2010 and revenues in the world market are forecasted to reach 1.62 billion dollar in 1999.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.61-64
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• 2001

Delta-integration vectors were constructed for the purpose of achieving homologous integration of the hirudin expression cassette into the chromosome of Saccharomyces cerevisiae. A double $\delta$ system truncated with the unnecessary bacterial genes, and consequently having a reduced insert size for integration, showed a four-fold increase in transformation efficiency at given DNA concentrations, and as a result, the constructed recombinant yeast strain had a 1.3-fold enhancement in hirudin expression level compared with a single $\delta$ system.

• Journal of Microbiology and Biotechnology
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• v.3 no.3
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• pp.161-167
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• 1993

Immobilized metal ion affinity chromatography (IMAC) was used to separate various types of recombinant hirudins from the culture broth. The wild type hirudin exhibited a retention in Cu(II)-chelated affinity chromatgoraphy since it contained a single exposed histidine at position 51. To obtain a stronger retention on an IDA-Cu(II) column, the hirudin variants were genetically engineered to contain one or two histidine (s) more than the wild type. While the affinity of the variants for IDA-Cu(II) ligand increased in comparison to that of the wild type, the antithrombin activities reduced to a certain degree. Cu(II), Ni(II) and Zn(II) ions were applied separately to the metal chelate column to investigate ligand specificity with respect to protein retention. As a result, the Cu(II) chelated chromatography gave the best resolution for all the hirudins tested and appeared to be the only IMAC that could be used generally for the purification of hirudins with a decreasing pH gradient.

• Journal of Chest Surgery
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• v.35 no.6
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• pp.475-478
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• 2002

A 45 year old man was admitted for aggravated dyspnea, abdominal distension and poor oral intake. On Echocardiogram, mitral stenosis(severe), tricuspid regurgitaion(IV), and LA thrombus were diagnosed. We used heparin with continuous infusion for prevention of systemic thrombo embolism. On the 11th day of admission, the patient showed thrombocytopenia and we suspected Heparin-induced thrombocytopenia. Hirudin was used in this case as alternative anticoagulant during cardiopulmonary bypass to prevent serious complication of heparin. The patient was recovered without any complication as postoperative bleeding or systemic thromboembolism.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.11a
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• pp.121-127
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• 1996

• Korean Journal of Head & Neck Oncology
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• v.16 no.1
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• pp.20-25
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• 2000

The medicinal leech, Hirudo medicinalis, has been used for salvage of the venous-congested flap following reconstructive surgery, with increasing frequency during the last two decades. Medicinal leech therapy is a safe, efficacious, economical, and well-tolerated intervention. The flap salvage with leeching occurs in approximately 70% of cases, and leeches need to be used as early as possible. At the time of the bite, evacuation of blood and injection of the leech saliva products, including anticoagulants and inhibitors of platelet aggregation, maintain capillary circulation of the flap, and then venous capillary return is established across the wound by angiogenesis. One of the salivary products, hirudin, represents the first parenteral anticoagulant introduced since the discovery of heparin. We analyzed two cases in which flap salvage with leeching was attempted, and reviewed medicinal leech therapy on the basis of our experiences and literature review. This has not been documented in the head and neck surgery literature in our country.