• 한국환경농학회지
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• 제28권4호
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• pp.435-441
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• 2009

In the present study, we analyzed the defensin protein deduced from Korean radish (Raphanus sativus L.) seeds.To express the genes in E. coli, we constructed a recombinant expression vector with a defensin gene, named rKRs-AFP gene isolated from Korean radish seeds. Over expressed rKRs-AFP proteins was separated by SDS-PAGE to determine the purity, and protein concentration was determined by the Bradford method. Antifungal activity was assessed by disk assay method against the tested fungi. As a result, when 500 mL of cell culture were disrupted by sonicator, 32.5 mg total proteins were obtained. The purified protein showed a single band on SDS-PAGE with estimated molecular weight about 6 KDa, consistent with the molecular mass calculated from the deduced amino acid sequence. The purified rKRs-AFP protein showed remarkable antifungal activities against several fungi including Aspergillus niger, Botrytis cinerea causing the gray mold disease, and Candida albicans. In field tests using the purified rKRs-AFP protein, the protein showed the reducing activity of disease spot and the mitigating effect of spreading of disease like agrichemicals. The immuno-assay of rKRs-AFP protein showed that the purified protein entirely accumulated at B. cinerea cytoplasm through the hyphal septa shown by fluorescence imaging. There was no fluorescence inside the cell, when the hypha was incubated without the protein. These all results indicate that the recombinant rKRs-AFP proteins can be utilized as a potential antifungal drug to control harmful plant fungal pathogens.

• BMB Reports
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• 제29권3호
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• pp.192-199
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• 1996

A rapid and sensitive method has been developed to detect a-fetoprotein (AFP) in human serum by a two-site sandwich enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies (MAbs) for human AFP within 1 h. To obtain the most sensitive and reliable MAbs. 12 kinds of MAbs (HPJ1 to HPJ12) as a capture antibody and 4 kinds of horseradish peroxidase (HRP) conjugated MAbs as a tracer antibody were investigated. Among these, only HPJ 10-HRP conjugated HPJ 1 (HPJ 10-HPJ $1^*$) and HPJ 11-HRP conjugated HPJ 10 (HPJ 11-HPJ $10^*$) were chosen as candidates based on the linearity of the standard curve and the sensitivity of the assay. To further characterize these two pairs. MAbs against human AFP were purified from hybridoma cells. conjugated with HRP. and then characterized to optimize the two-site sandwich ELISA The HPJ 10-HPJ $1^*$ pair showed a sensitivity of 1 ng/ml and a better reproducibility than the HPJ 11-HPJ $10^*$ pair when the human sera were incubated at $37^{\circ}C$ for 30 min. The results obtained for 480 randomly selected human sera showed 0~20 ng/ml of AFP values for the normal human sera. To test the utility of our kit, AFP concentrations were determined for 951 human sera (including 85 normal sera, 480 random blood sera, 213 HBsAg-positives. 50 anti-HCV antibody positives. and 47 malignant diseases) and compared with other commercially available AFP detecting kits. These results show that the present two-site sandwich ELISA method is a rapid, sensitive, and reliable procedure for detecting AFP in human serum.

• 한국수정란이식학회지
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• 제31권4호
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• pp.355-365
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• 2016

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

• The Korean Journal of Physiology and Pharmacology
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• 제12권4호
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• pp.193-197
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• 2008

The influence of alpha-fetoprotein (AFP) on the bone marrow (BM) natural suppressor (NS) cells of intact Ehrlich carcinoma -bearing CBA mice was studied. Bone marrow NS cells were fractionated into three fractions by isopycnic centrifugation on percoll gradients: NS1 (${\rho}$=1.080 g/ml), NS2 (${\rho}$=1.090 g/ml) and NS3 (1.100> ${\rho}$ > 1.090 g/ml). These fractions were highly different in their sensitivity to known NS cell inductors (interleukin (IL)-2, IL-3 or histamine). None of the NS fractions isolated from the intact mice spontaneously produced antiproliferative activity, however, they showed a high level of NS (antiproliferative and natural killer cell inhibitory) activity under the influence of AFP. A single injection of AFP to intact mice led to an increase of spontaneous NS activity and the inhibition of natural killer cell activity. NS activity, especially NS2, was increased in when tumor cells were subcutaneously inoculated three days after AFP injection. In the AFP-treated mice, the tumor mass at 14 days was 60% larger than that in the untreated mice. Our data confirmed that AFP is a tumor marker that can inhibit cancer immunity and plays a role in cancer pathogenesis.

• 원예과학기술지
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• 제31권6호
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• pp.720-725
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• 2013

수축성이 강한 코이어더스트가 주요 구성물질로 조제된 혼합상토는 충전량에 따라 용기내의 밀도가 달라지고 결과적으로 공극률(TP), 기상률(AFP) 및 용기용수량(CC)의 상대적인 비율이 변화된다. 따라서 충전량 차이에 따라 변화된 상토의 물리성 변화를 구명하여 작물 재배를 위한 기초자료로 제시하고자 본 연구를 수행하였다. 연구 목적을 달성하기 위해 코이어더스트 + 팽연왕겨(8:2, v/v; coir dust + expanded rice hull, CD + ERH로 지칭), 코이어더스트 + 훈탄(6:4; coir dust + carbonized rice hull, CD + CRH), 그리고 코이어더스트 + 분쇄된 수피(8:2; coir dusts + ground and raw pine bark, CD + GRPB)의 세 종류 상토를 조제하였다. 이 후 직경이 6.0, 7.5, 8.5, 10.5 및 12.5cm의 포트에 최적 가비중을 기초로 90, 100, 110, 120 및 130%가 되도록 상토 충전량을 조절한 후 TP, AFP 및 CC 변화를 측정하였다. 모든 규격의 포트에서 상토 충전량이 많아질수록 TP가 감소하였지만 CD + GRPB 상토보다 CD + ERH 또는 CD + CRH 상토의 감소 정도가 컸다. 상토 충전량이 많아질 경우 CC도 감소하였지만, 동일한 충전 비율에서 포트의 크기가 커질수록 CC의 감소한 정도가 컸다. 상토 충전량이 많아질수록 AFP도 뚜렷하게 감소하였다. 그러나 포트의 직경이 7.5cm보다 적을 때에는 동일한 충전 비율에서 CD + CRH 상토의 AFP가 가장 높았고, 포트의 크기가 8.5cm 이상으로 커질 경우 CD + ERH 상토의 AFP가 가장 높았다. 본 연구에서 상토 충전량 증가는 CC보다 AFP에 더 심한 영향을 미쳤으며 근권부의 가스 교환이 작물 생육에 큰 영향을 미침을 고려할 때 AFP의 감소를 방지할 수 있도록 충전량을 조절해야 한다고 판단하였다. 본 연구 결과는 상토를 이용한 용기재배에서 충전량 차이에 따른 물리성 변화를 예측하기 위한 자료로 활용될 수 있을 것이다.

• 유기물자원화
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• 제23권4호
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• pp.52-62
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• 2015

본 연구에서는 퇴비화공정에서 첨가재로 주로 사용되는 톱밥을 대상으로 톱밥 내 공기투과성 변화에 영향을 미치고 있는 요인별 특성과 총차감압력에 미치는 기여도를 평가하였다. 공기투과성 변화에의 영향요소로서 층류속도(v), 난류속도($v^2$), 수분함량(MC*v), 공기공극률(AFP*v), 입자크기(SIZE*v)를 선정하여 차감압력 산정을 위한 회귀식을 제시하였다. 차감압력에의 증가요인은 층류속도(v)와 입자크기(SIZE*v)이며, 감소요인은 난류속도($v^2$), 수분함량(MC*v), 공기공극률(AFP*v)이다. 공기유입속도를 높이면 총차감압력이 지속적으로 증가하였으며, 이러한 총차감압력 변화에 영향을 크게 미치는 증가요인은 입자크기(SIZE*v)이며, 감소요인은 공기공극률(AFP*v)이다. 또한 총차감압력에의 증가기여도는 낮은 유속에서는 층류속도(v)가 높은 유속에서는 입자크기(SIZE*v)의 기여도가 높았으며, 감소기여도는 공기공극률(AFP*v)이 가장 높았다. 반면에 수분함량 변화에 따른 총차감압력 변화는 그리 크지 않았다. 따라서 총차감압력은 증가요인보다는 감소요인인 공기공극률(AFP*v)과 수분함량(MC*v)에 의하여 영향을 받고 있음을 알 수 있다. 본 연구결과로서, 효율적 퇴비화공정을 위하여는 적정 수분함량 유지와 함께 공기공극률이 높은 첨가재를 선정하여 공기투과성을 향상시키는 방법이 적절할 것이다.

• 한국자기공명학회논문지
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• 제26권4호
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• pp.46-50
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• 2022

Antifreeze proteins (AFPs) bind to the ice crystals and then are able to inhibit the freezing of body fluid at subzero temperatures. Type III AFPs are categorized into three subgroups, QAE1, QAE2, and SP isoforms, based on differences in their isoelectric points. We prepared the QAE2 (AFP11) and SP (AFP6) isoforms of the notched-fin eelpout AFP and their mutant constructs and determined their temperature gradients of amide proton chemical shifts (𝚫δ/𝚫T) using NMR. The nfeAFP11 (QAE2) has the distinct 𝚫δ/𝚫T pattern of the first 3₁₀ helix compared to the QAE1 isoforms. The nfeAFP6 (SP) has the deviated 𝚫δ/𝚫T values of many residues, indicating its backbone conformational distortion. The study suggests the distortion in the H-bonding interactions and backbone conformation that is important for TH activities.

• 한국식물병리학회지
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• 제14권3호
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• pp.203-208
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• 1998

We isolated and characterized an antifungal peptide from the seeds of Phytolacca americana. Growth inhibition assay with Botrytis cinerea was used to screen inhibitory proteins from 60 different plant species. A 4 kDa antifungal peptide (Pa-AFP) inhibitory to hyphal growth of B. cinerea was found in the seeds of P. americana. The peptide Pa-AFP was purified to homogeneity by chromatographies of Sephadex G-50, DEAE-Sepharose, Sephacryl S-300, and C18 reverse-phase HPLC. Western blot analysis showed that a polyclonal antibody raised against the purified peptide cross-reacted with a 4 kDa protein in seeds but not in root and leaf tissues of P. americana. Pa-AFP inhibited the hyphal growth of Botrytis cinerea, Rihzoctonia solani, Fusarium oxysporum, and Magnaporthe grisea. Pa-AFP exhibited growth inhibition of Saccharomyces cerevisiae strain BWG7a, which was sensitive to osmotin.

• 보험의학회지
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• 제25권
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• pp.3-7
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• 2006

• 한국임상수의학회지
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• 제27권2호
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• pp.175-178
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• 2010

5년령의 수컷 코커스파니엘견이 갑작스런 발작증상으로 내원하였다. 내원당시 중등도의 탈수, 과다유연이 관찰되었고, 의식이 소실된 상태였다. 신체검사상 상복부에서 복강내 종괴가 촉진되었고, 실험실 검사에서 저혈당증과 간효소 수치의 상승이 관찰되었다. 진단으로는 병력, 임상증상, 신체검사, 신경검사, 기본적 실험실 검사와 방사선 검사 및 세포검사가 이용되었다. 특히 원발성 간 선암종이 의심되어 혈청 중 alpha-fetoprotein (AFP)을 측정하였다. 혈청 중 AFP 수치는 특이적으로 정상견에 비해 매우 높은 수준이었다(1513.5 ng/ml). 이 환축은 원발성 간 선암종이 의심이 되었으며 이로 인한 저혈당으로 발작증상을 관리하기 위해 포도당투여를 포함한 약물치료가 시작되었다. 하지만, 보호자의 요청에 의해 안락사가 실시되었고 부검 후 조직 검사를 통하여 원발성 간 선암종으로 확진되었다. 결론적으로 본증례의 경우 개에서 원발성 간암의 진단에 혈청 중 AFP 측정이 진단에 유용할 수 있음을 보여주는 증례다.