• 제목/요약/키워드: iron binding

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Direct Action of Relaxin on the Pig Mammary Glands to Promote Development of the Alveoli and Remodeling of the Extracellular Matrix (Relaxin Hormone의 직접젇인 작용에 의한 돼지 유선상피세포의 발달과 세포외간 기질의 분해개조 촉진)

  • Min, G.S.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.585-592
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    • 2004
  • Our previous studies demonstrated that relaxin in concert with estrogen promotes development of the mammary parenchyma during the last third of gestation in gilts, and the specific relaxin-binding sites were present in the mammary gland. This study was conducted to determine if relaxin-binding sites in the mammary gland were functional relaxin receptors. Three cycling cross-bred gilts were bilaterally ovariectomized on day 0 of the experiment. Beginning on day 15 and continuing through day 29 post-surgery, the gilt received an im. injection of estradiol benzoate at 12-hr intervals. Beginning on day 22 post-surgery, highly purified porcine relaxin was administered(lug/hr) into the left fourth mammary gland from the anterior end via miniature osmotic pump. Physiological saline was administered to the right fourth mammary gland. The gilt was sacrificed on day 29 post-surgery and histological characteristics of the mammary parenchyma were examined. The mammary glands treated locally with saline showed little, if any, lobulo-alveolar development, whereas the mammary glands treated with relaxin showed not only marked lobulo-alveolar development but also prominent secretions in the alveoli. The saline-treated glands were characterized by relatively dense and highly organized collagen fiber bundles. Whereas, in the relaxin-treated mammary glands, collagen fiber bundles were dispersed and loosely organized. In conclusion, relaxin-binding sites in the mammary gland are functional relaxin receptors and relaxin acts directly on the pig mammary gland to promote development of the alveoli and remodeling of the extracellular matrix.

Expression of human lactoferrin N-lobe in Pichia pastoris and its antibacterial activity (Pichia pastoris에서 사람 락토페린 N-lobe의 발현과 항균활성)

  • Won, Su-Jin;Jo, Jae-Hyung;Kim, Seung-Hwan;Kwon, Hyuk-Jin;Lee, Hyune-Hwan
    • Korean Journal of Microbiology
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    • v.51 no.3
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    • pp.271-279
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    • 2015
  • Lactoferrin (LF) is a multifunctional, iron-binding glycoprotein found in physiological secretions of mammals. LF shows antibacterial, antiviral and antifungal activities. In the present study, a gene encoding the N-terminal lobe of human lactoferrin (hLF) was isolated, cloned and expressed in methylotrophic yeast, Pichia pastoris. The recombinant hLF-N (rhLF-N) protein was secreted into the culture medium at the level of $458{\mu}g/ml$ in 3 L fermentor. The size of purified hLF-N was estimated as 35 kDa when analyzed by SDS-PAGE and western blotting. The rhLF-N was further confirmed by immunodiffusion using the anti-hLF polyclonal antibody. The expression profile analysis by qRT-PCR showed that the relative mRNA expression of rhLF-N was maximal after 2-3 days of methanol induction and reduced gradually at 4 days. The purified rhLF-N showed broad antibacterial activities against the pathogens such as Staphylococcus aureus, E. coli, Pseudomonas aeruginosa, Burkholderia cepacia, and Salmonella typhimurium. However, rhLF-N showed relatively lower activity when compared to peptides derived from LF. In spite of this weak activity, the rhLF-N expressed in P. pastoris might be more advantageous for the industrial application, because rhLF-N is secreted into the culture medium and the production can also be increased by optimization of culture conditions.

Characterization of lactoferrin hydrolysates on inflammatory cytokine expression in Raw264.7 macrophages

  • Son, Ji Yoon;Park, Young W.;Renchinkhand, Gereltuya;Paik, Seung-Hee;Nam, Myoung Soo
    • Korean Journal of Agricultural Science
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    • v.45 no.3
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    • pp.437-446
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    • 2018
  • Lactoferrin is an iron-binding glycoprotein which is present in colostrum, milk, and other body secretions. Lactoferrin activities are associated with inflammatory and immune responses. The aim of this study was to investigate the effect of lactoferrin hydrolysates (LH) on the production of immunomodulatory factors such as inflammatory related cytokines (tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, interleukin (IL)-6, and interleukin (IL)-13) in Raw264.7 cells, which originated from murine macrophages. The results show that the Raw264.7 cells cultured in 3 types (whole, and above and below 10 kDa) of lactoferrin hydrolysates (LH) did not show any cytotoxicity in the cells. $TNF-{\alpha}$ decreased dose-dependently to 1,500 - 2,000 ng/mL by treatment with the 3 types of LH at 1, 50, $100{\mu}g/mL$, whereas the positive control, lipopolysaccharide (LPS), and negative control produced 2,450 and 1,000 ng/mL of $TNF-{\alpha}$, respectively, in the Raw264.7 cells. The treatment with the 3 types of LH (whole and above and below 10 kDa) at $50{\mu}g/mL$ produced about 20 - 28 ng/mL of $IL-1{\beta}$ at 3, 6, and 9 h, respectively, while the negative control produced 7 ng/mL, and LPS as the positive control produced 48 - 60 ng/mL. $TNF-{\alpha}$ and IL-6 expression was decreased dose-dependently by the 3 types of LH. The mRNA levels of IL-13 were slightly increased dose-dependently by the whole and above 10 kDa LH, but decreased dose-dependently by the below 10 kDa LH in the Raw264.7 cells. The results show that LH had immunomodulating effects on cytokine production in anti- and pro-inflammatory reactions as well as anti-allergic reactions.

Optimization of the Expression of the Ferritin Protein Gene in Pleurotus eryngii and Its Biological Activity (큰느타리버섯에서 석충 페리틴 단백질 유전자의 발현 최적화 및 생물학적 활성)

  • Woo, Yean Jeong;Oh, Si Yoon;Choi, Jang Won
    • The Korean Journal of Mycology
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    • v.47 no.4
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    • pp.359-371
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    • 2019
  • To optimize the expression and secretion of ferritin protein associated with ion storage in the mushroom, Pleurotus eryngii, a recombinant secretion vector, harboring the ferritin gene, was constructed using a pPEVPR1b vector under the control of the CaMV 35S promoter and signal sequence of pathogen related protein (PR1b). The ferritin gene was isolated from the T-Fer vector following digestion with EcoRI and HindIII. The gene was then introduced into the pPEVPR1b secretion vector, and it was then named pPEVPR1b-Fer. The recombinant vector was transferred into P. eryngii via Agrobacterium tumefaciens-mediated transformation. The transformants were selected on MCM medium supplemented with kanamycin and its expression was confirmed by SDS-PAGE and western blotting. Expression of ferritin protein was optimized by modifying the culture conditions such as incubation time and temperature in batch and 20 L airlift type fermenter. The optimal conditions for ferritin production were achieved at 25℃ and after incubating for 8 days on MCM medium. The amount of ferritin protein was 2.4 mg/g mycelia, as measured by a quantitative protein assay. However, the signal sequence of PR1b (32 amino acids) seems to be correctly processed by peptidase and ferritin protein may be targeted in the apoplast region of mycelia, and it might not be secreted in the culture medium. The iron binding activity was confirmed by Perls' staining in a 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in P. eryngii mycelia. Mycelium powder containing ferritin was tested as a feed additive in broilers. The addition of ferritin powder stimulated the growth of young broilers and improved their feed efficiency and production index.

A Protective Effect of Chlorella Supplementation on Cadmium-induced Nephrotoxicity in the Rats

  • Hwang Yoo-Kyeong;Choi Hyun-Jin;Nan Meng;Yoo Jai-Du;Kim Yong-Ho
    • Biomedical Science Letters
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    • v.12 no.1
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    • pp.29-33
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    • 2006
  • The uptake of cadmium in animals is mainly accumulated in and affected to the liver and kidney by binding with red blood cells and serum albumin. The process accounts for more than 50% of the total accumulated cadmium in the body. The kidneys may be damaged without regarding the pathway uptake of cadmium. In a group of rats on supplements of 1% chlorella and 40 ppm cadmium, the concentration of cadmium in urine greatly decreased by 66% compared to control group, and the total synthesis of metallothionein decreased by 48.6% compared to control group. However, no previous study has assessed the protective effect on kidney damage induced by cadmium uptake through supplementation with chlorella. This study analyzed the biochemical marker for kidney damage in the rats after uptake of 40 ppm $CdCl_2$ and supplementation of the diet of Sprague Dawley (SD) rats with 1%, 5%, and 10% chlorella during 4 weeks. In a group of SD rats on supplementation with 1% chlorella and uptake of 40 ppm $CdCl_2,\;\beta_2$ microglobulin in the urine was found to be $3.1\pm0.6\;{\mu}g/L$, a decrease of 58% compared to a group of Sp rats on uptake of $CdCl_2$ only, in which the $\beta_2$ microglobulin was found to be $4.9\pm0.7\;{\mu}g/L$. According to the results of histopathological observation, the accumulation of mild and localized chronic inflammatory cells in kidney tissues was observed in 50% of the SD rats on uptake of cadmium only. In contrast, only 30% of the SD rats on supplementation with 1% chlorella and uptake of 40ppm $CdCl_2$, representing a histopathological abnormality, and there were no histopathological abnormalities at all in groups of SD rats on supplementation with 5% or 10% chlorella and uptake of 40 ppm $CdCl_2$. In conclusion, protein, calcium, and iron, which account for more than 50% of the total dried chlorella composition, may contribute to the reduction nephrotoxicity by stimulating both inhibited absorption of cadium and increased excretion of accumulated cadmium in kidneys.

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FACTORS AFFECTING THE PRODUCTION OF SULFUR COMPOUNDS BY FUSOBACTERIUM NUCLEATUM (Fusobacterium nucleatum의 유황화합물 생성에 영향을 미치는 인자)

  • Oh, In-Gyun;Park, Eun-Hae;Oh, Jong-Suk;Yang, Kyu-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.33-40
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    • 2003
  • Fusobacterium nucleatum, one of the bacteria causing halitosis, produces the volatile sulfur compounds (VSC) such as $H_2S$ in the media containing sulfur components, and forms FeS by binding with iron component. The various factors of oral cavity affect the concentration of sulfur compounds produced by Fusobacterium nucleatum. In this study, the effect of nutrients and pH on the production of sulfur compounds by Fusobacterium nucleatum was studied with the following results. 1. The optical density of broth was increased to $0.817{\pm}0.032$ and $1.297{\pm}0.024$ by adding 1.0% sodium thiosulfate and 0.05% L-cysteine hydrochloride in the media, respectively. 2. Though the optical density of broth was $0.799{\pm}0.032$ by adding volatile sulfur compounds (VSC) only in the media, it was increased to $1.775{\pm}0.003$ and $1.648{\pm}0.022$ by adding xylitol combined with glucose and fructose, respectively. 3. The concentration of VSC was above 20,000 ppb in the media above pH 5.5. The optical density of broth was still high in the media with L-cysteine hydrochloride of higher concentration, being low in the media of lower pH. 4. The concentration of VSC was high when there was distilled water or saline solution on the media, and their amount was small. These results suggest that the production of sulfur compounds by Fusobacterium nucleatum was inhibited by xylitol and acid.

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Effect of lactoferrin hydrolysates on inflammatory cytokine modulation in HEK-293, RBL-2H3, and HMC-1 cells

  • Son, Ji Yoon;Bae, Hyung Churl;Renchinkhand, Gereltuya;Nam, Myoung Soo;Kim, Woan-sub
    • Korean Journal of Agricultural Science
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    • v.47 no.1
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    • pp.83-93
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    • 2020
  • Lactoferrin (LF) is an iron-binding glycoprotein that is present in colostrum, milk, and other body secretions. The objective of this study was to investigate the effects of lactoferrin hydrolysates (LHs) on the production of immunomodulatory factors, including inflammatory related cytokines. The nuclear factor (NF)-κB reporter assay using human embryonic kidney 293 cells (HEK-293) revealed that NF-κB activity was significantly decreased by 1, 50, and 100 ㎍/mL of LH and the fractions above and below the 10 kDa LH. The mRNA expression of interferon (IFN)-γ in rat basophilic leukemia mast cells (RBL-2H3) treated with the fraction above the 10 kDa LH decreased in a dose-dependent manner, but the cells treated with LH and the fraction below the 10 kDa LH showed an increased expression of IFN-γ in a dose-dependent manner. The level of cyclooxygenase (COX)-2 expression decreased dose-dependently in RBL-2H3 cells treated with LH and the fraction above the 10 kDa LH, but the cells treated with the fraction below the 10 kDa LH showed an increased COX-2 expression in a dose-dependent manner. The mRNA expression of interleukin (IL)-4) was dose-dependently decreased by the fraction below the 10 kDa LH in human mast cells (HMC-1). The mRNA expressions of tumor necrosis factor (TNF)-α and IL-6 were significantly dose-dependently decreased by the fractions above and below the 10 kDa LH, but was dose-dependently increased by LH. The production of IL-4 was a little increased by the fraction above the 10 kDa LH compared to the positive control, but was decreased with LH and the fraction below the 10 kDa LH in HMC-1 cells. It was concluded that LF hydrolysates had an immunomodulating effect on anti-, pro-inflammatory and anti-allergic reactions.

Transgenic Siberian Ginseng Cultured Cells That Produce High Levels of Human Lactoferrin (인체 락토페린 생산 형질전환 가시오갈피 배양세포)

  • Jo Seung-Hyun;Kwon Suk-Yoon;Kim Jae-Whune;Lee Ki-Teak;Kwak Sang-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.209-215
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    • 2005
  • Lactoferrin is an iron-binding glycoprotein with many biological roles, including the protection against microbial and virus infection, stimulation of the immune system. We developed the transgenic Siberian ginseng (Acanthopanax senticosus) cell cultures producing the human lactoferrin (hLf) protein following Agrobacterium tumefaciens-mediated transformation. A construct containing a targeting signal peptide from tobacco endoplasmic reticulum fused to hLf cDNA under the control of an oxidative stress-inducible SWPA2 promoter was engineered. Transgenic Siberian ginseng cultured cells to produce a recombinant hLf protein were successfully generated and confirmed by PCR and Southern blot analysis. ELISA and western blot analysis showed that full length-hLf protein was synthesized in the transgenic cells. The production of hLf increased proportionally to cell growth and reached a maximal (up to 3% of total soluble proteins) at the stationary phase. These results suggest that the transgenic Siberian ginseng cultured cells in this study will be biotechnologically useful for the commercial production of medicinal plant cell cultures to produce hLf protein.

Respiratory Health of Foundry Workers Exposed to Binding Resin (RESIN 취급 주물공장 근로자들의 호흡기 건강에 관한 연구)

  • Choi, Jung-Keun;Rhee, Chang-Ok;Paek, Do-Myung;Choi, Byung-Soon;Shin, Yong-Chul;Chung, Ho-Keun
    • Journal of Preventive Medicine and Public Health
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    • v.27 no.2 s.46
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    • pp.274-285
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    • 1994
  • The effects of resin on the respiratory health have been investigated in 309 workers from four iron and steel foundries and the results compared with those from 122 workers who were not significantly exposed to resin gas and silica dust at the same industries. Phenol-formaldehyde resin was used in the core making and molding processes and workers were exposed to their decomposition products as well as to silica dust containing particulates. The subjects were grouped according to formaldehyde, dust and other gas exposures, and smoking habits were considered also in thi analysis. Standardized respiratory symptom questionnaire was administered by trained interviewers. Chest radiograph, pulmonary funtion tests, and methacholine challenge tests were done. Environmental measurements at the breathing zone were carried out to determine levels of formaldehyde, respiable dust and total dust. Foundry workers had a higher prevalence of symptoms of chronic bronchitis with chronic phlegm and chronic cough when exposed to dust. Exposure to gas was significantly associated with lowered $FEV_1$ and obstructive pulmonary function changes. Exposure to formaldehyde and phenol gas was associated with wheezing symptom among workers, but $FEV_1$ changes after methacholine challenge were not significantly different among different exposure groups. When asthma was defined as the presence of bronchial hyperreactivity with more than 20% decrease in $FEV_1$ after methacholine challenge, 17 workers out of 222 tested had asthma. Fewer asthmatic welters were found among groups exposed to formaldehyde, gas and dust, which indicates a healthy worker effects ill a cross-sectional study. The concentration of formaldehyde gas ranged from 0.24 to 0.43 ppm among studied foundries. The authors conclude that formaldehyde and phenol gas from combusted resin is probably the cause of asthmatic symptoms and also a selection force of those with higher bronchial reactivity away from exposures.

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Effects of Alloying Elements and Binding Materials on the Corrosion Behavior of Metal Hydride Electrodes (금속수소화물전극의 부식특성에 미치는 합금원소와 결합제의 영향)

  • Lee, Yang-Boum;Choe, Han-Cheol;Park, Ji-Yoon;Kim, Kwan-Hyu
    • Transactions of the Korean hydrogen and new energy society
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    • v.9 no.4
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    • pp.161-167
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    • 1998
  • It has been investigated the effects of alloying elements and binders on the corrosion behavior of metal hydride electrodes for anode of Ni/MH secondary battery. The $AB_5$-type alloys, $(LM)Ni_{4.49}Co_{0.1}Mn_{0.205}Al_{0.205}$ and $(LM)Ni_{3.6}Co_{0.7}Mn_{0.3}Al_{0.4}$, were used for the experiments. The electrodes were prepared by mixing and cold-pressing of alloy powders with Si sealent or PTFE powders, or cold-pressing the electroless copper coated alloy powders. The amount of copper coating was 20wt%. In order to examine corrosion behavior of the electrodes, the corrosion current and the current density, in 6M KOH aqueous solution after removal of oxygen in the solution, were measured by potentiodynamic and cyclic voltamo methods. The results showed that Co in the alloy increased corrosion resistance of the electrode whereas Ni decreased the stability of the electrode during the charge-discharge cycles. The electrode used Si sealant as a binder showed a lower corrosion current density than the electrode used PTFE and the electrode used Cu-coated alloy powders showed the best corrosion resistance.

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