• 한국미생물·생명공학회지
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• 제19권5호
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• pp.477-486
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• 1991

Aspergillus niger가 생산하는 $\alpha$-galactosidase의 효소학적 성질을 조사하기 위하여 시험균주를 밀기울 배양한 후 생성된 $\alpha$-galactosidase를 염석, 이온교환 크로마토그래피 및 겔 여과 등의 방법으로 정제한 후 정제효소의 효소학적 성질을 검토하였다. Asp. niger를 밀기울 배지에서 $30^{\circ}C$, 4일 배양했을 때 효소활성이 가장 높았으며 $\alpha$-galactosidase는 황산암모늄 염석, DEAE-cellulose 및 DEAE-Sephadex A-50 이온교환 크로마토그래피, sephadex G-150 겔 여과 등에 의하여 23.7배까지 정제되었으며 비활성이 1,229U/mg.protein, 수율 14이었고 HPLC와 PAGE에 의해 순도가 확인되었다.

• 미생물학회지
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• 제27권4호
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• pp.348-356
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• 1989

PIeurotus ostrealus의 배양액에서 syringic acid에 의해 유도되는 peroxidase의 활성이 추정되었다. 본 효소는 DEAE Sephadex A-50 이온교환크로마토그래피와 Sephadex G-150 첼 여과크로마토 그래피를 통하여 순수분리 되었다. 순수분리된 효소는 당함량이 35.7%인 당단백젤이었으며. SDS-linear polyacrylamide gradient gel 전기영동과 젤여과크로마토그래피에 의해 분자량(Mr) 72.400인 2개의 동일한 소단위체로 이루어진 이합체로 판명되었다. 본 효소는 흡광 스펙프럼의 분석결과, iron protoporphyrin IX의 구조를 갖는 2개의 heme을 조효소로 가지고 였는 것으로 추정되였다. 본 효소의 등전점은 4.26 이었고 $H_2O_2$에 대한 $K_m$값은 $7.2{\mu}M$이었다. 본 효소의 반응 최적온도는 $40^{\circ}C$, 최적 pH는 3.5에서 4.0 사이였다. Ferulic acid와 sinapic acid에 대한 본 효소의 친화도(Km)는 horseradish peroxidase에 비해 각각 2.4. 12.3배 더 높은 것으호 계산되었다.

• Journal of Microbiology and Biotechnology
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• 제16권10호
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• pp.1544-1553
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• 2006

The halophilic enterobacteria, Enterobacteria cancerogenus, was isolated from the intestines of the fusiform fish (Trachurus japonicus) to yield a protein-like material termed PLM-f74. PLM-f74 was characterized by strong inhibition ratios to angiogenesis (82.8% at the concentration of $18.5{\mu}g/ml$) and elevated antioxidative capacities with low toxicity. The PLM-f74 is a glycoprotein comprised of saccharides and amino acids. PLM-f74 inhibited cell adhesion that non-activated U937 monocytic cell adhesion to HUVECs activated with $IL-1{\beta}$ by 78.0%, and the adherence of U937 cells treated with the PLM-f74 and stimulated with $IL-1{\beta}$ to unstimulated HUVECs decreased by 102%. When both cell types were pretreated with PLM-f74, the adhesion of U937 cells to $IL-1{\beta}$-stimulated HUVECs was completely suppressed by 121% at a concentration of $18.5{\mu}g/ml$. PLM-f74 blocked signal pathways from VEGFR2, PI3K, ${\beta}$-catenin, and VE-cadherin to NF-kB, based on western bolt analysis. It also inhibited IL-l-stimulated HUVEC expression of the adhesion molecules, ICAM-l by 40%, VCAM-l by 60%, and E-selectin by 70% at the same concentration noted above. New anti-angiogenic and anti-cell adhesion materials showing elevated antioxidative capacities, and non-toxicity may be expected from these results.

• Journal of Korean Neurosurgical Society
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• 제60권5호
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• pp.518-526
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• 2017

Objective : Thromboembolism is the one of the most serious complications that can occur during endovascular coil embolization of cerebral aneurysm. We report on the effectiveness and safety of intra-arterial/intravenous (IA/IV) glycoprotein IIb/IIIa inhibitor (tirofiban) infusion for treating thromboembolism during endovascular coil embolization of cerebral aneurysm. Methods : We performed a retrospective analysis of 242 patients with ruptured or unruptured cerebral aneurysms (n=264) who underwent endovascular coil embolization from January 2011 to June 2014. Thromboembolism occurred in 20 patients (7.4%), including 14 cases of ruptured aneurysms and 6 cases of unruptured aneurysms. The most common site of aneurysms was the anterior communicating artery (n=8), followed by middle cerebral artery (n=6). When we found an enlarged thromboembolism during coil embolization, we tried to dissolve it using tirofiban administered via IA and IV loading ($5{\mu}g/kg$, respectively) for 3-5 minutes followed by IV maintenance ($0.08{\mu}g/kg/min$) for approximately 4-24 hours. Results : In 4 of 5 patients with total vessel occlusion, the vessel was recanalized to Thrombolysis in Cerebral Infarction Perfusion Scale (TICI) grade 3, and in 1 patient to TICI grade 2a. In 2 patients with partial vessel occlusion and 13 patients with minimal occlusion, the vessel recanalized to TICI grade 3. Irrelevant intracerebral hemorrhage was noted in 1 patient (5%), and thromboemboli-related cerebral infarction developed in 5 patients (25%), of which only 1 (5%) was symptomatic. Conclusion : IA/IV infusion and IV maintenance with tirofiban appear to be an effective rescue treatment for thromboembolism during endovascular coil embolization in patients with ruptured or unruptured cerebral aneurysms.

• Infection and chemotherapy
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• 제50권4호
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• pp.311-318
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• 2018

Background: Zoster vaccination is recommended for people with a history of herpes zoster (HZ), but the most effective timing of vaccine administration after zoster illness is unresolved. This prospective observational study compared the immunogenicity and safety of administering HZ vaccine at 6-12 months and 1-5 years after zoster illness. Materials and Methods: Blood samples were collected before the administration of live zoster vaccine and 6 weeks after vaccination. Varicella-zoster virus (VZV) IgG concentrations and T-cell responses were assessed by glycoprotein enzyme-linked immunosorbent assay and interferon-${\gamma}$ enzyme-linked immunospot assay (ELISPOT), respectively. Results: The baseline geometric mean value (GMV) of VZV IgG was higher in the 6-12 months group than in the 1-5 years group (245.5 IU/mL vs. 125.9 IU/mL; P = 0.021). However, the GMV increased significantly in both groups (P = 0.002 in the 6-12 months group; P <0.001 in the 1-5 years group). The results of the ELISPOT assay were not significant for differences of the GMV between baseline and 6-week post-vaccination groups, while the GMV increased significantly in both groups (P = 0.001 in the 6-12 months group; P <0.001 in the 1-5 years group). Conclusion: The immunogenicity of zoster vaccine may be similar whether administered 6-12 months, or >1 year after zoster illness. Trial Registration: ClinicalTrials.gov Identifier: NCT02704572

• Biomolecules & Therapeutics
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• 제30권6호
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• pp.546-552
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• 2022

Epidermal cell adhesion molecule (EpCAM) is a tumor-associated antigen (TAA), which has been considered as a cancer vaccine candidate. The EpCAM protein fused to the fragment crystallizable region of immunoglobulin G (IgG) tagged with KDEL endoplasmic reticulum (ER) retention signal (EpCAM-FcK) has been successfully expressed in transgenic tobacco (Nicotiana tabacum cv. Xanthi) and purified from the plant leaf. In this study, we investigated the ability of the plant-derived EpCAM-FcK (EpCAM-FcK^P) to elicit an immune response in vivo. The animal group injected with the EpCAM-FcK^P showed a higher differentiated germinal center (GC) B cell population (~9%) compared with the animal group injected with the recombinant rhEpCAM-Fc chimera (EpCAM-Fc^M). The animal group injected with EpCAM-FcK^P (~42%) had more differentiated T follicular helper cells (Tfh) than the animal group injected with EpCAM-Fc^M (~7%). This study demonstrated that the plant-derived EpCAM-FcK fusion antigenic protein induced a humoral immune response in mice.

• 생명과학회지
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• 제21권8호
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• pp.1156-1162
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• 2011

Equine herpesvirus type 3 (EHV-3)는 말[마(馬)]에서 말구진을 유발하고 말산업에 치명적인 악영향을 끼치는 매우 중요한 병원체이다. 국내에서 처음으로 말구진이 발생하였고, 말 생산농가에 경제적 피해를 입혔으나 다행히 원인바이러스 분리에 성공하였다. 이에 원인바이러스의 생물학적 특징을 연구하여 다음과 같은 결과를 얻었다. EHV-3를 RK-13 cell에 접종한 결과 접종 48 시간부터 세포변성효과가 나타남을 확인하였고 접종 72 시간부터는 세포박리가 일어났다. 세포를 수확하여 바이러스 DNA를 추출한 후 PCR 검사를 수행하였다. 그 결과 EHV-3에만 양성이었고 EHV-1 및 EHV-4에는 음성이었다. Swab에서 바이러스 DNA를 추출하여 10배씩 serial dilution을 한 후 PCR 검사를 실시한 결과 $10^4$까지 밴드를 확인할 수 있었다. 국내분리 EHV-3의 gG 특정부위에 대한 염기에서 표준주인 EHV-3 334/74 strain과 397개가 일치하여 99.25%의 상동성을 나타내었고 국내분리주를 EHV-3 거로주(Georo strain)라 명명하였다. 전기영동 결과에서는 EHV-3가 EHV-1보다 적은 수의 밴드를 확인할 수 있었다. 국내 분리주 EHV-3 거로주의 단백질을 분석한 결과 145 kD, 60 kD, 45 kD, 40 kD에서 단백 밴드를 관찰하였다.

• 대한수의학회지
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• 제39권2호
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• pp.247-256
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• 1999

A disease of young Holstein calves characterized by recurrent pneumonia, ulcerative and granulomatous stomatitis, enteritis with bacterial overgrowth, periodontitis, delayed wound healing, persistent neutrophilia and death at an early age had been originally described in 1983 and again in 1987. Most of these calves had stunted growth and a persistent, progressive neutrophilia (often exceeding 100,000/ml). By investigation of pedigrees, all of the affected calves have now been traced to a common sire and confirmed by polymerase chain reaction (PCR) diagnostic DNA testing to be homozygous carriers of a defective allele for bovine CD18. Neutrophils from these calves have several functional deficits and, most importantly, fail to adhere in a ${\beta}_2$-integrin dependent manner. The ${\beta}_2$-integrins represent a family of glycoproteins which participate in various leukocyte adhesion reactions during host defense. The presence or absence of ${\beta}_2$-integrin molecules can be demonstrated on the surface of neutrophils, monocytes and lymphocytes from normal or affected calves using specific monoclonal antibodies and flow cytometry, or by colloidal gold immunolabeling and scanning electron microscopy in backscatter mode. Deficiency of the ${\beta}_2$-integrins on all leukocyte types in Holstein calves is analogous to leukocyte adhesion deficiency (LAD) seen in humans. Neutrophils in bovine (BLAD) and human LAD patients are unable to adhere to the endothelial lining of the cardiovascular system thus interrupting egression of neutrophils into infected tissues. Other leukocytes, while still deficient in expression of the ${\beta}_2$-integrins, are still able to efficiently egress from the blood stream due to interactions of other adhesion molecules that are not as highly expressed on neutrophils. Both BLAD cattle and LAD children (who do not receive bone marrow transplants) often die at an early age as a result of the failure of neutrophils to extravasate into infected tissues. In 1991, Shuster, et $al^{27}$, identified two point mutations within the alleles encoding bovine CD18 in a Holstein calf afflicted with leukocyte adhesion deficiency. One mutation causes an aspartic acid to glycine substitution at amino acid 128 (D128G) in an extracellular region of this adhesion glycoprotein that is highly conserved (> 95% identity) between humans, cattle and mice. The other mutation is silent. Numerous calves with clinical symptoms of leukocyte adhesion deficiency have since been tested and all have been found homozygous for the D128G allele. In addition, calves homozygous far the D128G allele have been identified during widespread DNA testing in the United States. All cattle with the mutant allele are related to one bull, who through artificial insemination (A.I.), sired many calves in the 1950's and 1960's. The carrier frequency of the D128G CD18 allele among U.S. Holstein cattle had reached approximately 15% among active A.I. bulls and 8% among cows. By 1993, the organization of the dairy industry and the diagnostic test developed to genotype cattle, enabled virtually complete eradication of bovine leukocyte adhesion deficiency among current and future A.I. bulls.

• 한국동물학회지
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• 제26권3호
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• pp.155-170
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• 1983

溫熱處理에 대해서 細胞가 反應하여 適應하는 過程을 細胞膜水準에서 연구하기 위해서 培養中인 纖維芽細胞性 腫瘍細胞를 이용하여 細胞週期, 細胞密度 및 溫熱處理에 따른 膜表面蛋白質의 變化를 lactoperoxidase를 이용한 iodination과 galactose oxidase를 이용한 tritiation 方法을 서서 分析하였다. 細胞週期와 細胞密度에 따라 膜表面蛋白質은 定量的 變化를 보였는데 $G_1$期에서는 LETS 蛋白質과 高分子蛋白質이 크게 增加하였고 細胞密度의 증가에 따라서는 125K 蛋白質의 增加와 130K 및 100K 蛋白質의 減少가 特異하게 나타났다. 溫熱處理후의 시간경과에 따른 변화를 보면 處理직후 80K 이상의 蛋白質은 모두 사라지고, 24시간 지나면 70K 蛋白質이 현저한 增加를 보였지만 48시간이 경과하면 다시 減少하고 高分子蛋白質들이 原狀으로 回復되었다. 또한, 溫度를 $39^\\circ\\sim45^\\circC$까지 증가시켜 보았을 때 70K 蛋白質이 $41^\\circC$에서 가장 큰 폭으로 增加하는 特異한 현상이 관찰되었다. 아울러 이 70K 단백질은 trypsin을 처리하면 사라졌는데 galactose oxidase로 tritiation하였을때도 iodination하였을 때와 동일한 變化樣相을 보였다. 이러한 결과와 細胞質蛋白質과 比較한 缺課로 미루어 볼 때, 70K 蛋白質은 膜表面蛋白質이며 糖蛋白質이고 또한 HSP 70과 같은 蛋白質인 것으로 推定되었다. 이 蛋白質의 細胞膜에 있어서의 가능한 機能과 腫瘍細胞가 正常細胞에 비해서 溫熱處理에 敏感한 原因등에 관해서 考察하였다.

• Parasites, Hosts and Diseases
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• 제53권4호
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• pp.395-402
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• 2015

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.