• Korean Journal of Environmental Agriculture
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• v.28 no.4
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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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• v.29 no.3
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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.

• Journal of Embryo Transfer
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• v.31 no.4
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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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• v.12 no.4
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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.

• Horticultural Science & Technology
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• v.31 no.6
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• pp.720-725
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• 2013

When highly shrinkable materials such as coir dust are major component of root media, the degrees of compaction during container filling of root media severely influences the physical properties of root media. It results in the changes in total porosity (TP), container capacity (CC) and air-filled porosity (AFP). This research was conducted to secure the fundamental information in changes of soil physical properties as influenced by the compaction of root media during container filling. To achieve this, three root media were formulated by blending coir dust (CD) with expanded rice hull (CD + ERH, 8:2, v/v), carbonized rice hull (CD + CRH, 6:4) and ground and raw pine bark (CD + GRPB, 8:2). Based on the optimum bulk density, the amount of root media filled into 6.0, 7.5, 8.5, 10.5 and 12.5 cm were adjusted to 90, 100, 110, 120 and 130% based on the weight of root media. Then the changes in TP, CC, and AFP were measured. Elevation of the packing amount of root media in all sizes of pot resulted in the decrease of TP. But the decrease was more severe in CD + ERH and CD + CRH than those in CD + GRPB. The CC also decreased gradually as the packing amounts were elevated in three root media, but the decreases were severe as the container sizes became larger. The AFP decreased drastically by the elevation of the packing amount of root media in all sizes of pot. The AFP was the highest in CD + CRH medium when pot sizes were smaller than 7 cm, but that was the highest in CD + ERH when the pot sizes were larger than 8.5 cm among the 3 root media tested. In this research, the elevation of packing amount of three root media influenced more severely the AFP rather than CC. This result indicates that the packing amount should be controlled to maintain appropriate level of AFP because AFP rather than CC influence severely crop growth. The results obtained through this study can be used to predict the changes in physical properties of root media as influenced by packing amount in various sizes of pots.

• Journal of the Korea Organic Resources Recycling Association
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• v.23 no.4
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• pp.52-62
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• 2015

This study estimated the contribution rates of inlet air flow, moisture content, air-filled porosity and particle size on the total pressure drop for the sawdust used as the bulking agent in the composting. The statistical model for pressure drop including the affecting factors was proposed.($R^2=0.998{\sim}0.950$) While the laminar air flow(v) and particle size(SIZE*v) had the positive relations to the total pressure drop, the turbulent air flow($v^2$), moisture content(MC*v) and air-filled porosity(AFP*v) had the negative relations. Total pressure drop sharply increased with increasing of the inlet air flow. And the most significant factors affecting to total pressure drop were the particle size(SIZE*v) as positive factor and air-filled porosity(AFP*v) as negative factor. The contribution rate to total pressure drop by the particle size(SIZE*v) was continuously increased with increasing of the inlet air flow, but the contribution rate by air-filled porosity(AFP*v) was decreased. And total pressure drop was little changed even though the increasing of moisture content above the range of dry moisture content 0.25. The contribution rates of affecting factors had the different tendencies with increasing of the moisture content, especially in the negative factors as air-filled porosity(AFP*v) and moisture content(MC*v). For effective composting process, it is preferable to select the sawdust with higher air-filled porosity as bulking agent to enhance the air permeability.

• Journal of the Korean Magnetic Resonance Society
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• v.26 no.4
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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.

• Korean Journal Plant Pathology
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• v.14 no.3
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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.

• The Journal of the Korean life insurance medical association
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• v.25
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• pp.3-7
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• 2006

• Journal of Veterinary Clinics
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• v.27 no.2
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• pp.175-178
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• 2010

A 5-year-old male Cocker spaniel was presented with an abrupt seizure episode. The dog was moderately dehydrated, hypersalivation and loss of consciousness at the presentation. Tentative diagnosis was made through history taking, clinical signs, physical examination, neurologic examination, basic laboratory examinations, radiological examination and cytologic examination. In addition, serum alpha-fetoprotein (AFP) value was measured to diagnose in this case. The serum AFP concentration was marked elevated (1513.5 ng/ml). The patient was suspected as HCC and medical therapy was initiated to control hypoglycemic-associated seizure. However, the dog was euthanized because of the owner's request. It was diagnosed as hepatocellular carcinoma (HCC) based on the histopathologic examination. This case indicates that AFP measurement might be valuable to diagnose HCC in dogs.