• Journal of Microbiology and Biotechnology
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• 제17권8호
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• pp.1271-1283
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• 2007

A fibrinolytic protease (PoFE) was purified from the cultured mycelia of the edible oyster mushroom Pleurotus ostreatus, using a combination of various chromatographies. The purification protocol resulted in an 876-fold purification of the enzyme, with a final yield of 6.5%. The apparent molecular mass of the purified enzyme was estimated to be 32 kDa by SDS-PAGE, fibrin-zymography, and size exclusion using FPLC. The optimal reaction pH value and temperature were pH 6.5 and $35^{\circ}C$, respectively. PoFE effectively hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$-chain and the $B{\beta}$-chain over the ${\gamma}$-chain. Enzyme activity was enhanced by the addition of $Ca^{2+},\;Zn^{2+},\;and\;Mg^{2+}$ ions. Furthermore, PoFE activity was potently inhibited by EDTA, and it was found to exhibit a higher specificity for the chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 19 amino acid residues of the N-terminal sequence were ALRKGGAAALNIYSVGFTS, which is extremely similar to the metalloprotease purified from the fruiting body of P. ostreatus. In addition, we cloned the PoFE protein, encoding gene, and its nucleotide sequence was determined. The cDNA of cloned PoFE is 867 nucleotides long and consists of an open reading frame encoding 288 amino acid residues. Its cDNA showed a high degree of homology with PoMEP from P. ostreatus fruiting body. The mycelia of P. ostreatus may thus represent a potential source of new therapeutic agents to treat thrombosis.

• Journal of Microbiology
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• 제44권6호
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• pp.622-631
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• 2006

In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9 %. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and $37^{\circ}C$, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin $\alpha$-chain followed by the $\gamma$-$\gamma$ chains. It also hydrolyzed the $\beta$-chain, but more slowly. The A$\alpha$, B$\beta$, and $\gamma$ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by $Cu^{2+}$ and $Co^{2+}$, but enhanced by the additions of $Ca^{2+}$ and $Mg^{2+}$ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it's a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.

• 한국임상수의학회지
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• 제24권4호
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• pp.537-541
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• 2007

The purpose of this study is to examine and compare the concentrations of blood retinol and $\alpha$-tocopherol in normal and diarrhea calves. The subjects of the experiments are from three groups. Each group consists of 5 calves(diarrhea, treatment and control group). The CBC, serum chemistry, serum retinol and a-tocopherol concentrations were estimated in each group. Blood retinol concentration was $13.3{\pm}7.0{\mu}g/100ml$ in diarrhea group, $31.5{\pm}6.9{\mu}g/100ml$ in treatment group and $28.1{\pm}11.8{\mu}g/100ml$ in the control group. The blood concentration in diarrhea group was significantly lower than that of the control group in the case of retinol(p<0.05). However, there were no significance between the treatment poop and the control group. The $\alpha$-tocopherol concentration in blood was $266.0{\pm}127.6{\mu}g/100ml$ in diarrhea group, $432.2{\pm}172.7{\mu}g/100ml$ in the treated group and $579.3{\pm}145.8{\mu}g/100ml$ in the control group. In the case of $\alpha$-tocopherol, the laboratory group were significantly lower than the control group(p<0.05), except for the treatment group. As in retinol concentration there were no significance between treatment group and the control group. In the test of CBC, PCV was significantly lower in the group with diarrhea than the control group(p<0.05). Fibrinogen concentrations in diarrhea calves were significantly higher than the treatment and control group. In conclusion, the blood retinol and $\alpha$-tocopherol concentration in diarrhea calves are lower than normal calves. Medication of retinol and $\alpha$-tocopherol on calves with diarrhea is recommended.

• Journal of Chest Surgery
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• 제29권9호
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• pp.989-995
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• 1996

체외순환을 이용한 개심술후 혈소판기능부전에 의한 비정상적인 출혈은 수술 사망율 및 이환율의 증가를 초래한다. 본 논문에서는 체외순환시 aprotinin을 투여하여 임상적 지혈 및 혈소판기능보존에 대한 효과를 관찰하기 위해 인공판막치 환술을 받은 40명의 환자를 대상으로 low-dose aprotinln 투여군 20명, 대조군 20명으로 나누어 그 효과를 비교 분석하였다. Aprotinin 투여군에서 대조군에 비해 약 40%의 출혈감소(622.0$\pm$ 186m1 versus 1021 $\pm$483.5ml, p<0.01) 및 약 63%의 혈색소 손실(14.7 $\pm$6.8g versus 39.7$\pm$ 16.4g, p<0.01)을 줄일 수 있었으며 약 70%의 수혈감 소(197.7$\pm$ 56.3ml versus 651.2 $\pm$ 147.5ml, p<0.01) 효과를 얻을 수 있었다. 또한 투여 군에서 혈소판 응집 능이 효과적으로 보존되었다(P<0.05). 체외순환중 activated clotting time이 투여 군에서 연장되어 heparin의 항응고작용에 대해 상승효과가 있었으며 aprotinin의 투여와 관련된 합병증은 없었다. 이상의 결과에서 체외순환을 이용한 개심술에 있어서 low-dose aprotinin의 투여는 술후 출혈량 및 혈 액과 혈액제제의 사용감소에 효과가 있으며 이는 혈소판응집능의 효과적 \ulcorner보존에 의한 것으로 사료된다.

• 대한수의학회지
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• 제27권2호
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• pp.321-330
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• 1987

In the endemic area of bovine piroplasmosis in Chonbuk Province, total of 486 heads of Holstein cow selected from 14 farms randomly were examined the infection status of piroplasma and their blood values. The results obtained were summarized as follows: 1. The etiological agent was only Theileria sergenti and none of the Babesia sp. infection. 2. All of the 486 heads of examined cow were infected with Theileria sergenti (100%). 3. In the initial examination of 11 farms in early summer, the means of blood values were as follows; RBC:$526{\pm}84(10^4/mm^3$), Ht:$27.9{\pm}2.8%$, SP:$7.5{\pm}0.6g/dl$, WBC: $11586{\pm}354/mm^3$, and Fibrinogen: $578.5{\pm}164.1mg/dl$. 4. In the second examinations in slimmer season, the blood values of examined cows had little varieties. But in the winter season, cows housed in the barn, the blood values were greatly increased; RBC: $601{\pm}77(10^4/mm^3)$, (F-value: 37.186**), Ht: $30.5{\pm}1.3%$(F-value: 15.626**) Hb: $12.2{\pm}1.3g/dl$(F-value: 5.899**), SP: $7.4{\pm}0.6g/dl$(F-value: 5.05**). 5. To determine the piroplasma infection in the many herds of cow, the measurement of Ht, Hb, and SP values etc. were more effective and helpful to find the weak cow because the examined process are not only to be easily carry out many samples but also to be done very simple and rapidly. 6. The indications of therapeutic standard values were stablished as follows; the less than 20% in the Ht value, below the 10g/dl in the Hb value, and less than 6g/dl in SP value. When the one among the therapeutic standard values, Ht, Hb, and SP was below the therapeutic standard value, the cow was treated with nutriments. When more than two among them were below the therapeutic standard, the cow was treated with Berenil, at once. In this way, there was no one head which represented clinical signs of piroplasmosis among 486 heads of cow at 14 farms during the year of 1986. 7. In a Korean native cattle infected with Thieleria sergenti seriously, the recovery of blood values was very prolonged by the treatment of Berenil only. But by the transfusion of 300ml blood collected from normal cow, the blood values were recovered rapidly.

• 생명과학회지
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• 제28권8호
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• pp.923-930
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• 2018

본 연구는 미생물을 이용하여 갈색거저리를 발효시킨 분말을 water, ethanol, methanol 용매별로 추출하여 다양한 실험을 행하였다. 균주는 3종의 유산균 Lactobacillus plantarum JBMI F3 (F3), Lactobacillus plantarum JBMI F5 (F5), Lactobacullus gasseri Ba9 (Ba9), 1종의 곰팡이 Aspergillus kawachii KCCM 32819 (Ak), 1종의 효모 Saccharomyces cerevisiae KACC 93023 (Sc), 1종의 바실러스 Bacillus subtilis KACC 91157 (Bs) 총 6종의 균주를 사용하였다. 각 균주를 이용한 발효 후 갈색거저리의 효능을 알아보기 위해 DPPH assay, 총 phenolic compound 및 Flavonoid 함량, Cu 환원력, Fibrinolytic activity, 효소-기질 활성 분석을 진행하였다. 그 결과, DPPH assay 결과, Water 추출물이 전반적으로 높은 항산화능을 보였고, 그 중 Bs 균주 발효 추출물이 우수한 라디컬 소거능을 보였다. 총 phenolic compound 및 Flavonoid의 경우, phenolic compound는 Ak발효군이 가장 함량이 많았고, Flavonoid는 Bs발효군이 가장 높았다. 환원력은 ethanol - Bs발효군에서 높은 수치를 확인하였고, Fibrinolytic activity 및 전분분해능은 water - Bs발효군에서 우수한 효과를 나타냈다. 위와 같은 결과를 토대로 발효 갈색거저리분말은, 기존 갈색거저리 분말보다 항산화능 및 생리활성 측면에서 효능이 상당부분 향상됨을 알 수 있었다. 그러므로 발효거저리분말은 식품산업전반에서 높은 이용가치를 증명하는 기초 연구자료로 활용될 것이다.

• Parasites, Hosts and Diseases
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• 제30권3호
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• pp.177-182
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• 1992

Saimiri 원숭이 21마리와 Aotus 원숭이 12마리에게 Plasmedium falciparum Indochina CDC-1충주(원숭이 순응 충주)를 접종시킨 후 3주간 혈소판과 혈액치를 경시적으로 관찰하였던 바, 이들 원숭이 (Saimiri sciureus boliviensis와 Aotus nancymai karyotype-1)는 접종 14일 후에 최고의 기생률, 호중구감소증, 임파구증가증 그리고 심한 혈색소혈증과 섬유소원의 퇴행성 산물(FDP's)치의 증가를 수반한 섬한 혈소판 감소성 빈혈증이 관찰되었다. Aotus에 있어서는 평균혈색소량(MCH)과 평균혈구용적(MCV)치의 감소와 기생률의 증가를 가져왔으나, Saimiri에 있어서는 기생률이 상승함에 따라서 이들 혈액치는 유의적 차이로 상승하는 결과를 가져왔다. 일반적으로 이들 두 원숭이에 있어서 혈소판의 감소 증세와 최고 기생률을 14잎 째 나타낸 후 곧 정상 혈액치로 회복되는 소견이 관찰된다. 특히 혈소판의 변화에 있어서 Aotus 원숭이는 다른 혈액치의 변화 그리고 기생률과 깊 은 상관관계를 갖고 있었으나, Saimiri에 있어서는 상관관계를 유지하지 못하였다. 결론적으로 말라리아 연구시 Aotus 원숭이가 Saimiri 원숭이 보다 우수한 품종임과 말라리아 진단과정에 있어서 혈소판의 회복률과 재생은 말라리아의 감염시기 판정에 유용한 자료로서 활용될 수 있음을 관찰하였으며, 말라리아 병원성 판정에 대한 실험에 있어서 이들 두 종류 원숭이의 혈액치 변화 특이성을 보고하는 바이다.

• Journal of Trauma and Injury
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• 제33권1호
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• pp.1-12
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• 2020

Purpose: Despite recent developments in the management of trauma patients in South Korea, a standardized system and guideline for trauma treatment are absent. Methods: Five guidelines were assessed using the Appraisal of Guidelines for Research and Evaluation II instrument. Results: Restrictive volume replacement must be used for patients experiencing shock from trauma until hemostasis is achieved (1B). The target systolic pressure for fluid resuscitation should be 80-90 mmHg in hypovolemic shock patients (1C). For patients with head trauma, the target pressure for fluid resuscitation should be 100-110 mmHg (2C). Isotonic crystalloid fluid is recommended for initially treating traumatic hypovolemic shock patients (1A). Hypothermia should be prevented in patients with severe trauma, and if hypothermia occurs, the body temperature should be increased without delay (1B). Acidemia must be corrected with an appropriate means of treatment for hypovolemic trauma patients (1B). When a large amount of transfusion is required for trauma patients in hypovolemic shock, a massive transfusion protocol (MTP) should be used (1B). The decision to implement MTP should be made based on hemodynamic status and initial responses to fluid resuscitation, not only the patient's initial condition (1B). The ratio of plasma to red blood cell concentration should be at least 1:2 for trauma patients requiring massive transfusion (1B). When a trauma patient is in life-threatening hypovolemic shock, vasopressors can be administered in addition to fluids and blood products (1B). Early administration of tranexamic acid is recommended in trauma patients who are actively bleeding or at high risk of hemorrhage (1B). For hypovolemic patients with coagulopathy non-responsive to primary therapy, the use of fibrinogen concentrate, cryoprecipitate, or recombinant factor VIIa can be considered (2C). Conclusions: This research presents Korea's first clinical practice guideline for patients with traumatic shock. This guideline will be revised with updated research every 5 years.

• 한국임상수의학회지
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• 제8권2호
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• pp.127-142
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• 1991

This study was carried out to investigate fatty liver in Korean black goats. Adult female goats were divided into 3 test groups(A, B and C). Group A and B of goats each received 3 test consecutive daily doses of DL-ethionine at 75mg/kg and 150mg/kg body weight, respectively. Group C of goats was given 3 consecutive doses of the compound every 48 hours at 150mg/kg body weight. The clinical symptoms, hematological values, serum chemical values and histopathological study of the liver were investigated in the test animals. The results obtained are as follows ; 1. Fatty liver were observed in every test animal. 2. Some clinical symptoms( anorexia, depression) were appeared from 1st day to 7th day after administration of the compound in every test animal. In addition to these symptoms, diarrhea and salivation were generally observed in test animals which were given the compound at 150mg/kg body weight. The degree of these symptoms was dose dependent. 3. There was no significant variations in total WBC counts and fibrinogen values in the blood of test goats. The PCV values were significantly increased on 5th day of dosing in group A and B of goats. 4. The total lipid value was not changed but the concentration of NEFA was significantly increased on 3rd day of dosing with the compound and returned to normal value after 10 days hereafter. The value of triglycerides was significantly increased on 1st day and returned to normal value on 3rd day of dosing. The value of cholesterol was significantly decreased on 3rd day and returned to normal value on 10th day after treatment. 5. Total protein level was decreased on 10th day of dosing in the groups of B and C, and billirubin level was significantly increased on 7th day of dosing in every test group and returned to normal level after 13th day of administration. 6. The activity of GGT in serum was not changed while the activities of SDH and AST were significantly increased in every test goat and those values were returned to normal after 10~13th day of trestment. 7 The 35K-protein fraction in serum was not detected by SDS-polyacrylamide gel electrophoresis, but this protein fraction was detected by the same method after treating the 21st and 22nd fraction which were obtained by column chromatography with Sephadex G-100. 8. The affected liver was congested and swollen on 3rd day, and yellowish brown in color and mottled appearance on 7th day of treatment. Histopathologically, fat droplets were common in the hepatocytes, this change was intensive on 7th day after treatment in group B and C. Hepatic cell necrosis was observed in some livers but this pathological change was disappeared and returned to normal after 13 days of treatment.

• Macromolecular Research
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• 제9권2호
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• pp.107-115
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• 2001

Poly(L-lactide-co-glycolide)(PLGA) was blended with poly[$\omega$-methacryloyloxyethyl phospho-rylcholine-co-ethylhexylmethacrylate (PMEH)] (PLGA/PMEH) to endow with new functionality i.e., to improve the cell-, tissue- and blood-compatibility. The characteristics of surface properties were investigated by measurement of contact angle goniometer, Fourier-transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR) and electron spectroscopy for chemical analysis (ESCA). NIH/3T3 fibroblast and bovine aortic endothelial cell were cultured on control and PLGA/PMEH surfaces for the evaluation of ceil attachment and proliferation in terms of surface functionality such as the concentration of phosphoryl-choline. Also, the behavior of platelet adhesion on PLGA/PMEH was observed in terms of the surface functionality. The contact angles on control and PLGA/PMEH surfaces decreased with increasing PMEH content from 75$^{\circ}$ to about 43$^{\circ}$. It was observed from the FTIR-ATR spectra that phosphorylcholine groups are gradually increased with increasing blended amount of MPC. The experimental P percent values from ESCA analysis were more 3.28∼7.4 times than that of the theoretical P percent for each blend films. These results clearly indicated that the MPC units were concentrated on the surface of PLGA/PMEH blend. The control and PLGA/PMEH films with 0.5 to 10.0 wt% concentration of PMEH were used to evaluate cell adhesion and growth in terms of phosphorylcholine functionality and wettability. Cell adhesion and growth on PLGA/PMEH surfaces were less active than those of control and both cell number decreased with increasing PMEH contents without the effect of surface wettability. It can be explained that the fibronectin adsorption decreased with an increase in the surface density of phosphorylcholine functional group. One can conclude the amount of the protein adsorption and the adhesion number of cells can be controlled and nonspecifically reduced by the introduction with phosphorylcholine group. Morphology of the adhered platelets on the PLGA/PMEH surface showed lower activating than control and the number of adhered platelets on the PLGA/PMEH sample decreased with increasing the phosphorylcholine contents. The amount of fibrinogen adsorbed on the PLGA/PMEH surface demonstrated that the phospholipid polar group played an important role in reducing protein adsorption on the surface. In conclusion, this surface modification technique might be effectively used PLGA film and scaffolds for controlling the adhesion and growth of cell and tissue, furthermore, blood compatibility of the PLGA was improved by blending of the MPC polymer for the application of tissue engineering fields.