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Monitoring trafficking and expression of hemagglutinin-tagged transient receptor potential melastatin 4 channel in mammalian cells

  • Eun Mi Hwang;Bo Hyun Lee;Eun Hye Byun;Soomin Lee;Dawon Kang;Dong Kun Lee;Min Seok Song;Seong-Geun Hong
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.4
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    • pp.417-426
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    • 2023
  • The TRPM4 gene encodes a Ca2+-activated monovalent cation channel called transient receptor potential melastatin 4 (TRPM4) that is expressed in various tissues. Dysregulation or abnormal expression of TRPM4 has been linked to a range of diseases. We introduced the hemagglutinin (HA) tag into the extracellular S6 loop of TRPM4, resulting in an HA-tagged version called TRPM4-HA. This TRPM4-HA was developed to investigate the purification, localization, and function of TRPM4 in different physiological and pathological conditions. TRPM4-HA was successfully expressed in the intact cell membrane and exhibited similar electrophysiological properties, such as the current-voltage relationship, rapid desensitization, and current size, compared to the wild-type TRPM4. The presence of the TRPM4 inhibitor 9-phenanthrol did not affect these properties. Furthermore, a wound-healing assay showed that TRPM4-HA induced cell proliferation and migration, similar to the native TRPM4. Co-expression of protein tyrosine phosphatase, non-receptor type 6 (PTPN6 or SHP1) with TRPM4-HA led to the translocation of TRPM4-HA to the cytosol. To investigate the interaction between PTPN6 and tyrosine residues of TRPM4 in enhancing channel activity, we generated four mutants in which tyrosine (Y) residues were substituted with phenylalanine (F) at the N-terminus of TRPM4. The YF mutants displayed properties and functions similar to TRPM4-HA, except for the Y256F mutant, which showed resistance to 9-phenanthrol, suggesting that Y256 may be involved in the binding site for 9-phenanthrol. Overall, the creation of HA-tagged TRPM4 provides researchers with a valuable tool to study the role of TRPM4 in different conditions and its potential interactions with other proteins, such as PTPN6.

Spatially-resolved Photoluminescence Studies on Intermixing Effect of InGaAs Quantum Dot Structures Formed by AlAs Wet Oxidation and Thermal Annealing (AlAs 습식산화와 열처리로 인한 InGaAs 양자점 레이저 구조의 Intermixing효과에 관한 공간 분해 광학적 특성)

  • Hwang J.S.;Kwon B.J.;Kwack H.S.;Choi J.W.;Choi Y.H.;Cho N.K.;Cheon H.S.;Cho W.C.;Song J.D.;Choi W.J.;Lee J.I.
    • Journal of the Korean Vacuum Society
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    • v.15 no.2
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    • pp.201-208
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    • 2006
  • Optical characteristics of InGaAs quantum dot (QD) laser structures with an Al native oxide (AlOx) layer as a current-blocking layer were studied by means of photoluminescence (PL), PL excitation, and spatially-resolved micro-PL techniques. The InGaAs QD samples were first grown by molecular-beam epitaxy (MBE), and then prepared by wet oxidation and thermal annealing techniques. For the InGaAs QD structures treated by the wet oxidation and thermal annealing processes, a broad PL emission due to the intermixing effect of the AlOx layer was observed at PL emission energy higher than that of the non-intermixed region. We observed a dominant InGaAs QD emission at about 1.1 eV in the non-oxide AlAs region, while InGaAs QD-related emissions at about 1.16 eV and $1.18{\sim}1.20eV$ were observed for the AlOx and the SiNx regions, respectively. We conclude that the intermixing effect of the InGaAs QD region under an AlOx layer is stronger than that of the InGaAs QD region under a non-oxided AlAs layer.

Influence of Personal Characteristics and Background Characteristics on Entrepreneurship and Perceived Business Performance in Entrepreneurs as Independent Business Owners of Network Marketing (네트워크 마케팅 독립사업자 창업가의 개인 특성, 배경 특성이 기업가정신과 인지된 경영성과에 미치는 영향)

  • Yoon, Hae Sook;Song, In Bang;Kim, Yeon Jong
    • Asia-Pacific Journal of Business Venturing and Entrepreneurship
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    • v.13 no.4
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    • pp.233-244
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    • 2018
  • The purpose of this study was to improve awareness of network marketing business in South Korea, which takes a quite negative view of network marketing, to raise awareness of "starting up a business" as independent business owners, to assist them in building their own subjecthood in network business and entrepreneurial identity, and ultimately to give some suggestions on how to improve their self-directed entrepreneurial competency and the quality of their business management. To make an empirical analysis of 121 independent business owners of Korean native network marketing, the personal and background variables of the entrepreneurs were selected as independent variables, and entrepreneurship was selected as a mediator variable. The selected dependent variables were financial and non-financial business performances. A multiple regression analysis was conducted by inputting the variables. The findings of the study were as follows: It produced an effect of more financial performance when the innovation of the independent business owners of network marketing was better, and this innovation was possible to have only when they improved in entrepreneurial efficacy and locus of control. In contrast, only authenticity had an effect on boosting non-financial business performance among the factors of entrepreneurship. To have authenticity, it's necessary to strengthen achievement needs and awareness of locus of control among the personal characteristics of the independent business owners of network marketing. Among the factors of entrepreneurship, there was better authenticity when their personal networks were broader among their background characteristics. In addition, self-efficacy that was one of personal characteristics made a direct contribution to the enhancement of non-financial performance. As a result, independent business owners of network marketing are required to hold a strong belief in their own business, to be active in business activities and to have a high efficacy as spontaneous principal agents, and they also are required to have more adventurous, planned, active and propulsive achievement needs that enable them to attain the goals of their business and keep moving forward. Besides, they should foster their locus-of-control competency that is to control, endure and be responsible for a variety of phenomena or incidents that they face, and there will be better financial performance or non-financial performance only when they demonstrate their self-belief and confidence and when they have faith in and conviction about themselves. For independent business owners of network marketing, a consumer-centered thinking that entails authenticity and trustworthiness and touches the hearts of customers is a more effective marketing strategy than an egocentric thinking as sellers when they approach customers. And they are expected to make progress in terms of the quality of business management when their business attachment or enthusiasm is great enough to match their own efficacy with entrepreneurial identity or strike a balance between them.

Seedling Emergence of Dry -seeded Rice under Different Sowing Depths and Irrigation Regimes (건답직파에서 파종심도와 관개조건에 따른 벼 품종들의 출아특성)

  • 이변우;명을재
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.40 no.1
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    • pp.59-68
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    • 1995
  • Investigated were the relationships between plumule elongation characteristics and seedling emergence of 46 varieties including native, improved and red rice varieties of Korea, and varieties from U.S.A., Italy, India, Japan under 1, 3, and 5cm deep sowing with irrigated and non-irrigated condition. Experiments were carried out in paddy field of sandy loam. There was heavy shower of 19.2mm on the next day of seeding and thereafter, clear and dry weather continued during the experiment period. Soil temperature averaged over 30 days after seeding was $16.4^{\circ}C$ at 3cm depth. Soil hardness increased linearly up to 2.5kg /$cm^2$ on the 14th day after seeding, on which date irrigated plot was irrigated through furrow, and up to 4kg / $cm^2$ on the 28th day in non-irrigated plot. Soil hardness dropped near to 0kg /$cm^2$ after irrigation and developed up to 2.5kg /$cm^2$ again by 28 days after seeding. Seedling emergence was higher in irrigated plots than non-irrigated plots at all seeding depths. Korean improved varieties were substantially lower in seedling emergence under non-irrigated condition of 1 cm deep sowing than those under irrigated condition. This poor seedling emergence resulted mainly from delayed emergence by exposing them to greater soil strength. Percent seedling emergence under irrigated and non-irrigated condition showed signifi-cant correlations at 3 and 5 cm deep sowing. Korean improved varieties belonged to the group of poor seedling emergence, and I taliconaverneco, Chinsura Boro and Weld Pally to best group under both irrigation conditions at 3 and 5cm deep sowing. Seedling emergence showed highly signifi-cant positive correlation with the plumule length of mesocotyl + 1st internode + incomplete leaf and of mesocotyl+coleoptile. Among the characters constituting plumule length, incomplete leaf length showed greatest positive correlation followed by coleoptile and mesocotyl under irrigated condition at 3 and 5 cm deep sowing, and highest correlation with mesocotyllength followed by first internode and incomplete leaf under non-irrigated condition. Days to 50% seedling emergence at 1 cm deep sowing with irrigation showed great varietal variation of 10 to 30 days, and showed high significant negative correlations with percent seedling emergence under both irrigation conditions except for 1 cm deep sowing with irrigation, Days to seedling emergence revealed sig-nificant negative correlations with plumule characters except 2nd internode, showing highest cor-relation with incomplete leaf length.

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Investigation of Original Landscaping in the Vicinity of Yongyun and Hwahongmun in Suwon Hwaseong (수원 화성 용연(龍淵)과 화홍문 일곽의 원형경관 탐색)

  • Rho, Jae-Hyun;Choi, Jong-Hee;Shin, Sang-Sup
    • Journal of the Korean Institute of Landscape Architecture
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    • v.38 no.2
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    • pp.94-108
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    • 2010
  • The purpose of this study is to provide data for the restoration of 'Yongyun(龍淵)' and 'Hwahongmun(華虹門)' through an investigation of the vicinity of their original landscapes at the time of construction of Hwaseong in Suwon and through tracing the transformative process of the environments of this vicinity. The results are as follows; As identified by 'Yongyunjung(龍淵亭)' and 'Yongdugak(龍頭閣)', other names of Yongyun, 'Banghwasuryujung(訪花隨柳亭)', which was built on 'Yongduam', is a facility whose place identity is highlighted with a sense of unity with Yongyun. The south lakefront of Yongyun, bordering Banghwasuryujung, has boundaries that make the best use of the natural geographical features of Yongduam while the current circumference of Yongyun is comparatively shorter than its original state. The size of 'Joongdo(中島)', however, seems to be an example of apparent over-design complement and reorganize 'Joongdo', which had been restored larger than its original state at the time of restoration in the 1970s. The depth of 'Yongyun' was created to be lower than the actual depth, without consideration for its initial depth, as soil was accumulated through continuous flooding after it was created. It is assumed that the original drains which were installed about 10m inside the lake were created facing the stream. As regards the planting environment, a circular planting of willows was made in the outskirts of 'Yongyun', except the 'Yongduam' which is a pure forest, and a mix of 'Pinus densiflora', shrubbery and deciduous broad leaf trees was planted in 'Joongdo'. Of the plants growing in the area of this study, plant species introduced to Korea after Hwaseong was constructed are found, most of which provide interest and attraction. The old pine trees growing in a group once grew in the castle areas of the vicinity even in the 1920s, the period of Japanese occupation, but they disappeared from the area in the aftermath of subsequent urban development and the Korean War. Although restored to the site, the number and space taken up by these trees are insignificant compared to those of the original environment. On the basis of these results, the following is considered necessary for the true restoration of the vicinity of 'Yongyun' and 'Hwahongmun': First, the grounds of 'Yongyun' should be dredged deeply enough to expose the bedrock and should be recreated in the rough outline of a half moon by extension to the southwest toward 'Yongduam' and 'Hwahongmun', and the size of 'Joongdo' should be significantly reduced. Secondly, considering that most plant species, except the pine trees and wild trees in 'Yongduam', are non-native plants introduced in order to provide such attractions such as the appreciation of scenic areas, they should be replaced with native species, mainly with the pine trees which were utilized during the construction of Hwaseong. The weeping willows planted in the 'middle-island' should be relocated to the outskirts of 'Yongyun', and replaced with pine trees as the major trees and maple trees or deciduous broadleaf trees to fill in the gaps. Thirdly, exotic species such as the 'Pinus rigida' planted in a group around 'Banghwasuryujung' and 'Bugammun' and 'Pinus strobus' planted in the vicinity of Hwahongmun' should be removed.

Vinegar Produced from Chrysanthemum zawadskii and Pearl Shell (구절초진주식초)

  • Ann, Yong-Geun;Oh, Moon-Hun;Lee, Byung-Yup
    • The Korean Journal of Food And Nutrition
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    • v.25 no.1
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    • pp.90-98
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    • 2012
  • With the addition of ethanol to wax gourd extract and by acetic fermentation, 5.0% acidity vinegar was produced. After putting 10% extract(10% concentration) of Chrysanthemum zawadskii in this, and by dissolving shell, Chrysanthemum zawadskii-pearl vinegar was produced. When a 1% of ark shell, oyster shell, or ear shell was added to wax gourd vinegar, 95.6~98.4% of the shell dissolved, and when a 2% content of shell was added, 97.2~98.4% was dissolved. The acidity of vinegar which dissolved 1% shell was pH 3.0~3.17, and the acidity of vinegar which dissolved 2% shell was pH 1.11~ 1.20. The pH values of vinegar which dissolved 1%, and 2% shell contents were 4.54~4.55, and 4.86~4.95, respectively. When 1% shell was dissolved, the acidity was higher than that of commercial vinegar, with a high pH value and low level of free acid. This shows that when Chrysanthemum zawadskii 1% is added during acetic acid fermentation, the inhibition was 44.4%, and 22.2% respectively. In this regard, Chrysanthemum zawadskii should be added after the fermentation of acetic acid. The calcium content of 1% shell vinegar is 0.4%, and that of 2% vinegar is 0.78%. Non-heated native wax gourd shows an angiotensin converting enzyme inhibition rate of 21.7%, an antioxidant activity of 5.23%, and a tyrosinase inhibition rate of 5.5%. In the case of heated-extracted wax gourd, the angiotensin converting enzyme inhibition rate was 16.1%, superoxide dismutase activity was 20.5%, antioxidant activity was 23.2%, and the tyrosinase inhibition rate was 7.1%. Also, in the case of Chrysanthemum zawadskii, the angiotensin converting enzyme inhibition rate was 28.8%, the xanthine oxidase inhibition rate was 28.2%, the superoxide dismutase activity was 14.5%, the antioxidant activity was 3.2%, and the tyrosinase inhibition rate was 9.2% Data also revealed that when a 10% sample of the heated-wax gourd extract was added to A549 human lung cancer epithelial cells of, the number of cancer cells declined by 80% in 72 hours, When a 10% native extract was added, the number of cells declined by, 74% in 48 hours, and when a heated-extract of Chrysanthemum zawadskii was added, 100% of the cells died after 72 hours.

Analysis of Hematologic Characteristics of Korean Native Stripped Cattle Chickso according to the Ages (성장 단계별 희소 한우(칡소)의 혈액학적 분석)

  • Kim, Hyun;Cho, Young Moo;Ko, Yeoung-Gyu;Kim, Nam-Tae;Kim, Sung Woo;Seong, Hwan-Hoo
    • Journal of Embryo Transfer
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    • v.29 no.3
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    • pp.313-319
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    • 2014
  • Investigations for hematologic values and the differential count of WBC for Korean indigenous cattle (KIC) and Stripped Cattle (SC) are rarely performed. Therefore, when the index of complete blood counts (CBC) analysis of KIC and SC were requested, it had many difficulties to make the results for blood condition since the standard hematologic values of KIC and SC are lacking. The objective of this study was to investigated the hematologic values and the differential count of WBC for blood of total 19 striped cattle (SC) and 187 KIC as a control by estimation analysis of hematologic characteristics. As a result, the mean values of RBC and platelet of KIC were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between KIC and SC of the same age (2~3 years) showed the statistical significance (P<0.05). Also, in the WBC of KIC, the mean values were decreased according to the age from $13.8{\times}10^3/{\mu}l$ under 1 year to $9.5{\times}10^3/{\mu}l$ over 5 years. In the differential count of WBC of KIC, it showed generally the rates of 46.2% lymphocyte and 36% segmented neutrophil. Additionally, in comparative analysis between pregnant and non-pregnant group of KIC, the mean values of Hg and HCT in pregnant group were significantly decreased(P<0.05). In conclusion, data obtained from this study may be valuable as a standard for interpretation of the results in hematologic analysis of KIC. Result of this study will be used for establishing reference range for hematologic analysis in SC.

Distyly and Population Size of Abeliophyllum distichum Nakai, an Endemic Plant in Korea (한국 특산식물 미선나무의 이화주성(Distyly) 및 개체군 크기)

  • So-Dam Kim;Ae-Ra Moon;Shin-Young Kwon;Seok-Min Yun;Hwi-Min Kim;Dong-Hyoung Lee;Sung-Won Son
    • Korean Journal of Environment and Ecology
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    • v.36 no.6
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    • pp.639-650
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    • 2022
  • Abeliophyllum distichum Nakai, a rare plant with distylous characteristics, is native to certain parts of the Korean Peninsula. It is registered on the IUCN Red List of Threatened Species as a globally endangered plant. This study was conducted to establish an appropriate local conservation management plan suitable for future A. distichum populations by comparing and analyzing the flowering characteristics and population size according to distyly based on the results of quantitative surveys in 14 regions, including 8 areas with native populations of A. distichum and 6 natural monument populations. The number of individuals appearing in each population group was surveyed, and the flowering individuals were identified by style as being either pin or thrum flower types as they were being examined and recorded on the site. In total, 13,130 individuals of A. distichum (7,003 flowering and 6,127 non-flowering individuals) were recorded, but the balance of the number of pin- and thrum-flowered individuals in each population was not significant (p<0.05), indicating an imbalanced state. In particular, the Yeongdong (YD) population was very disproportionate compared to other populations, suggesting that its genetic diversity was low and the possibility of inbreeding was high. The average flowering and fruiting rates by management unit were much higher in the natural monument populations (89.2% and 55.3%, respectively) than in the natural habitat populations (39.0% and 8.5%, respectively). It may be due to a difference in reproductive growth resulting from light inflow into the forest caused by the upper crown closure. The area of occupation (AOO) of A. distichum on the Korean Peninsula covered an area of 23,224.5 m2. Although the natural monument population was smaller than the natural habitat population, its density was higher, likely as a result of the periodic management of natural monument populations, where the installation of protective facilities in certain areas restricts population spread. Conservation of A. distichum populations requires removing the natural monument populations suspected of anthropogenic and genetic disturbances and expanding the conservation priority population by designating new protected areas. Although the habitats of natural monument populations are managed by the Cultural Heritage Administration and local governments, there are no agencies that are responsible for managing natural habitat populations. Therefore, institutional improvement in the overall management of A. distichum should be prioritized.

Effects of Yeast Culture Supplementation on Rice Straw Digestibility and Cellulolytic Bacterial Community in the Rumen (볏짚 조사료에 대한 효모 배양물 첨가가 반추위 소화율 및 섬유소 분해균의 군락 변화에 미치는 영향)

  • Sung, Ha Guyn
    • Journal of Animal Science and Technology
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    • v.55 no.1
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    • pp.41-49
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    • 2013
  • In vitro and in situ incubation studies were conducted to determine effects of yeast culture supplements (Saccharomyces cerevisiae) on cellulolytic bacterial function and fiber digestion in rice straw. In vitro dry matter digestibility of rice straw gradually increased according to supplemental levels of yeast culture (0.0, 0.2, 0.4, 0.6, 0.8 and 1.0%). Digestibility of rice straw started to increase apparently when yeast culture was added more than 0.6% level (p<0.05). Also, we reconfirmed that in vitro dry matter digestibility was significantly increased by 0.6% of yeast culture addition in 4% NaOH treated and non-treated rice straws (p<0.05). When in situ dry matter digestibility was tested in Korean native goats fed basal diet or experimental diet which contained 1.0% of yeast culture, the yeast culture feeding improved in situ dry matter digestibility in both 4% NaOH treated and non-treated rice straws (p<0.05). In case of real-time PCR monitoring cellulolytic bacterial function, the bacterial population attached on rice straw showed the increasing trends with higher level of yeast culture spraying on rice straw. F. succinogenes and R. flavefaciens were significantly increased in accordance to spraying levels of yeast culture (0.0, 0.1 and 0.3%) at both 12 and 24 hrs of in situ incubation (p<0.05). R. albus was significantly higher population in yeast culture spraying than non-soraying at 12 hrs of in situ incubation (p<0.05). These bacterial populations were showed the increasing trends with digestibility enhancement of rice straw according to the higher levels of yeast culture supplement. Overall, these results clearly suggest that the presence of yeast culture result in noticeable increase of rice straw digestion, which is modulated via good effect on cellulolytic bacterial attachment to fiber substrates.

Studies on the Antibacterial Activity of Enzymatic Hydrolyzates of Lactoferrin Derived from Bovine Colostrum (유우 Lactoferrin 효소가수분해물 항균 활성에 관한 연구)

  • Han, Su Yeon;Kim, Jong Woo
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.52-67
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    • 1998
  • The investigative research on the mammalian milk purely consisted of the physiological quality of lactoferrin was conducted to reveal the antimicrobial ativity of specifically functional foods with antibiotic characteristics as a basic data in food manufacturing. Bovine lactoferrin were isolated from raw milk samples, and was digested with pepsin, trypsin and chymotrypsin. It was necessary then to separate and purify lactoferrin from bovine raw milk, and in order to analyze the antimicrobial activity of the enzyme-treated bovine lactoferrin in their required quantitative fraction. Afterwards Escherichia coli and Staphylococcus aureus was incubated in it. It was that investigated to enzyme-treated fractions molecular weight and the peptide fragment with antimicrobial effect. 1. The purity of enzyme-treated bovine lactoferrin(BLF) was tested by SDS-PAGE. As a results of 12% SDS-PAGE assay, pepsin-treated LF did not exhibited band until if reaches 14 KDa, while trypsin and chymotrypsin treated LF, known to contain the non-digestive lactoferrin exhibited band at a molecular weight of 33 KDa. 2. Bovine lactoferrin was sucessfully purified through the use of Sephadex G-50 Column. In order to assay LF through the Sephadex G-50 column chromatography, the digestive bovine lactoferrin (BLFs) was eluted with a linear gradient of 0.05% Tris-HCl. When the gel-filtration analysis, pepsin, trypsin and chymotrypsin treatments of BLF fragments was showed 2, 3, and 2 peak, respectively. The results of the HPLC analysis confirmed that had a non-digestive lactoferrin receptor, and trypsin and chymotrypsin treated BLFs has an antimicrobial effect. 3. To measure the strength of the antimicrobial effect of enzyme treated lactoferrin it was compared to the antimicrobial activity taking place at the incubated Escherichia coli and Staphylococcus aureus. This might explain the resistance of the microorganisms for peptide fragment. The pepsin-treated of bovine lactoferrin was markedly reduced by incubation of the cells. Trypsin-treated of BLF was similar to chymotrypsin-treated of BLF. However, trypsin and chymotrypsin treatments of BLFs were showed the antimicrobial effect until eight hours incubation for native bovine lactoferrin. Therefore the enzyme-treated lactoferrin have an antimicrobial effect even non-digestive lactoferrin. 4. The digestive bovine lactoferrin fragments assay was carried out by the use of Sephadex G-50 column chromatography and SDS-PAGE. The pepsin and chymotrypsin-treated fragments has a low molecular weight and trypsin-treated lactoferrin was only showed a band. It was described that characteristics of digestive protein. It appeared that there may be a relation between virulence and resistance to enzyme-treated BLF.

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