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AICAR (5-aminoimidazole-4-carboxamide-1-β-D-ribonucleoside) Decreases Protein Synthesis in C2C12 Myotubes Cultured in High Glucose Media (근육세포 내 Glucose 농도와 AICAR에 의한 단백질 합성 저해)

  • Park, Chang-Seok;Kim, Jae-Hwan;Oh, Young-Kyoon;Kim, Kyoung-Hoon;Choi, Chang-Weon;Cho, Eun-Seok;Jeong, Yong-Dae;Park, Sung-Kwon
    • Journal of Animal Science and Technology
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    • v.54 no.5
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    • pp.369-373
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    • 2012
  • AMP-activated protein kinase (AMPK) maintains energy homeostasis in skeletal muscle. Nonetheless, its functional role on protein synthesis with different nutrient availability has not been elucidated. Therefore, the purpose of this study is to examine the effect of AMPK activity on protein content in C2C12 myotubes incubated with low (5 mM; LG) or high (25 mM; HG) glucose media. LG stimulated (p<0.05) AMPK and acetyl CoA carboxylase (ACC) activity compare to those in HG group. Total protein content was higher in myotubes cultured with HG than in those cultured with LG and further increased by AICAR (5-amino-1-${\beta}$-D-ribofuranosyl-imidazole-4-carboxamide). Myotubes cultured with HG showed 7.5% lower UbFL (Ubiquitin Firefly Luciferase)-to-SV40 (Simian virus40) ratio compared to those in LG. Glucose level did not change the phosphorylation level of mammalian target of rapamycin (mTOR). Interestingly, administration of AICAR significantly increased phosphorylation level of mTOR in myotubes cultured with LG but not in those with HG. Overall, this data indicate that AMPK activity and protein turnover are finely regulated in response to different glucose concentration.

Effect of Imipramine on the Contractility of Single Cells Isolated from Canine Detrusor (Imipramine이 배뇨근 세포의 수축성에 미치는 직접작용)

  • Huh, Chan-Wook;Lee, Kwang-Youn;Kim, Won-Joon
    • Journal of Yeungnam Medical Science
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    • v.11 no.2
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    • pp.293-302
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    • 1994
  • The objective of this study was to establish a good methodology to isolate single smooth muscle cells that are alive and respond properly to pharmacological agents. Canine urinary bladders were employed as the source of single cells, and acetylcholine, atropine and imipramine were used as indicators of pharmacological responsiveness. Imipramine, an antidepressant drug exhibited the anticholinergic and calcium antagonizing properties on rat detrusor muscle. To establish a control value for a further experiment to elucidate the mechanism of action of imipramine on detrusor muscle, we measured the concentration-response of single cells to acetylcholine in the presesnce of imipramine by length of the cells and compared the result with the response in the presence of atropine. Tiny chops of smooth muscle taken from anesthetized canine urinary bladder were incubated in collagenase solution at $36^{\circ}C$ for 17-20 minutes. The collagenase solution included collagenase 1.2 mg/ml, soybean tryspin inhibitor 0.08 mg/ml, bovine serum albumin 2% in 10 ml Krebs-Henseleit buffer solution aerated with a consistent breeze of 95/5% $O_2/CO_2$, to maintain the pH at 7.4. After washing with plain K-H solution on 450 mesh, cells were dissociated from the digested tissue for 12-15 minutes. Cell suspension was transfered in 5 ml test tubes and acetylcholine was added for the final concentration to be $10^{-14}M{\sim}10^{-9}M$. To find the optimal time to fix the cells to determine the contractile responses, 1% acrolein was added 5, 10, 20, 30, 60 and 120 seconds after the administration of ACh. The length of cells fixed by acrolein were measured by microscaler via CCTV camera on phaes-contrast microscope. The average length of 50 cells from a slide glass was taken as the value of a sample at the very concentration point. Single cells were isolated from canine detrusor. The length of untreated cells varied from 82 ${\mu}m$ to 94 ${\mu}m$. The maximal response to actylcholine $10^{-9}M$ was accomplished within 5 seconds of exposure, and the shortening was $19{\pm}3$%. Atropine reduced the contraction of the cells concentration-dependently. Imipramine which exerts a cholinergic blocking action on some smooth muscles also reduced the contraction concentration-dependently and by a similar pattern as atropine. These findings document that imipramine may exerts a cholinergic blocking activity in the single smooth muscle cells isolated from canine urinary bladder.

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Enhancement of Anticarcinogenic Potentials of Submerged-Liquid Culture of Agaricus blazei Murill on Mouse Ascites Cancer by Rice Hull (왕겨에 의한 신령버섯균사체 액체배양액의 생쥐 항복수암성 증가)

  • Kim, Young-S.;Jang, Wook-J.;Rakib, A.;Kwon, Jung-M.;Ahn, Chae-R.;Kim, So-Y.;Cho, Yong-U.;Ha, Young-K.;Kim, Jeong-O.;Ha, Yeong-L.
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1402-1408
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    • 2010
  • The effects of rice hull (RH) powder on the anticarcinogenic activity of submerged-liquid cultures of Agaricus blazei Murill (AB) were assessed for mouse ascites cancers induced by mouse Sarcoma S-180 (S-180) cancer cells. Optimal growth of AB mycelia in the basal liquid culture medium, containing soybean meal, was achieved by culturing at $25^{\circ}C$ for 5 days, when evaluated by $\beta$-glucan content, Brix, and mycelial weight, relative to other culture conditions. Hot-water extract (HWE) of the submergedliquid culture of AB mycelia grown at $25^{\circ}C$ for 5 days exhibited a stronger anticarcinogenic activity, relative to HWE from other culture conditions. No such effects were obtained from AB mycelial cultures by alternative temperature-controlling cultures. Both cytotoxicity for S-180 cells and anticarcinogenic potentials for mouse ascites cancer of the HWE from AB mycelia grown in the basal medium containing 1% RH powder for 5 days at $25^{\circ}C$ were significantly (p<0.05) enhanced, relative to HWE from the AB mycelia culture of the basal medium without RH powder. These results indicate that HWE of submerged-liquid culture of AB mycelia, incubated in media containing 1% RH powder at $25^{\circ}C$ for 5 days, enhanced anticarcinogenic activity against S-180 cell-induced mouse ascites cancer, and suggest that RH powder is an excellent ingredient for the improvement of the anticarcinogenic potentials of the submerged-liquid culture of mushroom mycelia.

Alteration of Anaerobic Bacteria and S. mutans Count in Oral Cavity after Occlusal Stabilization Appliance Use (교합안정장치 사용에 따른 구강 내 혐기성 세균과 S. mutans의 변화)

  • Byun, Jin-Seok;Suh, Bong-Jik
    • Journal of Oral Medicine and Pain
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    • v.32 no.4
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    • pp.375-381
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    • 2007
  • Occlusal stabilization appliance is one of the most common treatment option for management of temporomandibular disorders. It acts in oral cavity for several hours per day, and usually it will take at least 6 months to 2 years of total wearing periods to take a treatment goal. In the oral cavity, occlusal stabilization appliance, unintentional manner, is able to acts as a reservoir of bacteria and protect bacteria from saliva and oxygen. This condition is so favorable to many bacteria such as S. mutans and other anaerobes, usually have been reported as causative factors of dental caries, periodontal disease and oral malodor. In this study, we investigated anaerobic bacteria and S. mutans count before and after occlusal stabilization appliance use to evaluate the possible role of occlusal stabilization appliance as protector of these bacteria. Four men(average 27.5 years) wore maxillary occlusal stabilization appliance at each night(average 9 hours) for 5 days. we swabbed saliva-plaque mixed sample at 3 different site(maxillary left 2nd molar, maxillary left central incisor, mandibular left 2nd molar) before and after occlusal stabilization appliance use. Each samples were plated in (1) anaerobic blood agar medium, (2) selective S. mutans medium(MS-MUTV) and incubated in anaerobic chamber($CO^2$ 10%, $37^{\circ}C$) for 72 hours. Each bacterial colony forming unit(CFU) were counted with naked eyes. From obtained data, we can conclude as follows: 1. There was some changes about anaerobic bacteria and S. mutans count in oral cavity after occlusal stabilization appliance use. 2. The number of anaerobic bacteria was significantly increased at maxillary 2nd molar(P=0.003), maxillary central incisor(P=0.020) after occlusal stabilization appliance use compared with before. 3. Occlusal stabilization appliance use itself had indirect effect to increase the number of anaerobic bacteria at other uncovered opponent tooth site. 4. The number of S. mutans was significantly increased at maxillary 2nd molar(P=0.043), maxillary central incisor (P=0.049) after occlusal stabilization appliance use compared with before. 5. Occlusal stabilization appliance use itself had not any effect on the number of S. mutans at other uncovered opponent tooth site.

The effect of captafol on the hematological value, erythrocyte membrane and plasma biochemical value in vitro (시험관내에서 captafol이 혈액학, 적혈구막 및 혈장생화학치에 미치는 영향)

  • Kim, Sung-hoon;Hwang, Hwa-seon;Cha, Shin-woo;Han, Sang-seop;Roh, Jung-koo
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.507-511
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    • 1993
  • The effect of captafol on the hematological value, erythrocyte membrane and plasma biochemical value was investigated using blood of SPF Sprague-Dawley male rats in vitro. For the anticoagulations, we used 0.5mg of heparin per $10m{\ell}$ of blood from vena cava. Three con-centrations($0.1{\times}10^{-4}M$, $1{\times}10^{-3}M$ and $1{\times}10^{-2}M$) captafol in ethanol were added to the each $2.5m{\ell}$ blood so that the linal concentration of ethanol was 1%. The blood contained with each concentration of captafol was incubated at $37^{\circ}C$ C for 2 hours under 5% $CO_2$ gas The whole blood and plasma were examined for hematological vaJues, erythrocyte membrane damage and biochemical values, respectively. The results obtained were summarized as follows ; 1. The number of RBC in $1{\times}10^{-2}M$ group and the concentration of MCHC in $1{\times}10^{-3}M$ group were significantly (p<0.05) decreased and increased from that of control values, respectively. The percentage of Hct was significantly (p<0.05) decreased with dose-response. 2. Erythrocyte fragility rate of $1{\times}10^{-3}M$ and $1{\times}10^{-2}M$ group were significantly (p<0.01) increased from that control with dose response. 3. Potassium ion level of $1{\times}10^{-2}M$ was significantly (p<0.05) increased from that of control. 4. The concentration of total bilirubin in the $1{\times}10^{-3}M$ and $1{\times}10^{-2}M$ groups were significantly increased from that of control. The enzyme level of creatine kinase in $1{\times}10^{-2}M$ group was significanlty (p<0.05) decreased from control value.

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Bakanae Disease Reduction Effect by Use of Silicate Coated Seed in Wet Direct-Seeded Rice (규산코팅 벼 종자를 이용한 담수직파재배 시 벼 키다리병 경감효과)

  • Kang, Yang-Soon;Kim, Wan Joong;Kim, Yeon Ju;Jung, Ki-Hong;Choi, Ul-Su
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.61 no.1
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    • pp.9-16
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    • 2016
  • To investigate the effect of soluble silicate zeolite dressing of the rice against bakanae disease, field trial in reclaimed land and in vitro were carried out. The coated rice seeds (SCS) which were dressed with the mixture of 25% silicic acids (binder), and the zeolite (coating powder). In wet direct seeding, uniform scattering of rice seeds on the soil surface and the better seedling establishment were shown in SCS treatment plots. The incidence of bakanae disease began from the mid tillering stage toward the heading stage. Around heading stage, the ratio of infected tillers reached its highest point by 9.9% in non-SCS treatment plots. While, in SCS treatment plots, the ratio of infected tillers was no more than 0.01%. The vitality of the pathogenic fungi of bakanae disease in the SCS and non-SCS samples were assessed. Samples were incubated for one week keeping proper humidity at $30^{\circ}C$ after inoculated with panicles of infected rice plants from experimental field plots. In non-SCS treatment, pinkish colonies were formed on the grain surface of panicle of infected plants, and mycelium, macro-conidia and micro-conidia were developed actively inside part of infected grain inoculated. While in SCS treatment, micro-conidia and mycelium were not survived and the growth of macro-conidia, mycelia were greatly inhibited and withered. Based on the results, it is concluded that the environmental friendly control of bakanae disease by use of SCS is possible and soluble silicate can be applied as agents for replacement of seed disinfection.

Process Development of a Virally-Safe Acellular Bovine Amniotic Membrane for Biological Dressing (바이러스 안전성이 보증된 무세포 소 양막 생물창상피복재 제조 공정 개발)

  • Bae, Jung-Eun;Kim, Chang-Kyong;Kim, Sung-Po;Yang, Eun-Kyung;Kim, In-Seop
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.420-427
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    • 2010
  • A process for manufacturing virally-safe bovine amniotic membrane(BAM) has been developed for biological dressing. BAM was harvested from a healthy bovine placenta, and then the epithelium was removed. The remaining stromal layer was consecutively disinfected with 70% ethanol and 0.05% sodium hypochlorite. The stromal layer was incubated in a decellularization solution containing 0.25%(w/v) trypsin to remove the cellular components. The resulting acelluar BAM was lyophilized to preserve its biochemical and structural integrity. The BAM was packed and exposed to 25 kGy of gamma irradiation for sterilization purpose. Histological, electron microscopical, and biochemical observations showed that the acellualr BAM had intact structural integrity of three dimensional collagen fibers and contained several growth factors, accelerating wound healing, such as EGF (Epidermal growth factor), KGF (Keratinocyte growth factor), and FGF (Fibroblast growth factor). Bovine herpes virus (BHV), bovine viral diarrhoea virus (BVDV), bovine parainfluenza virus type 3 (BPIV-3), and bovine parvovirus (BPV) were chosen as the biological indicators for validation of viral safety of the acellular BAM. Samples from relevant stages of the production process were spiked with each virus and subjected to viral inactivation processes. Viruses were recovered from the samples and then titrated immediately. All the viruses tested were completely inactivated to undetectable levels within 1 h of 70% ethanol treatment. Enveloped viruses such as BHV, BVDV, and BPIV-3 were more effectively inactivated than BPV by 0.05% sodium hypochlorite treatment. BHV, BVDV, and BPIV-3 were completely inactivated to undetectable levels by 25 kGy of gamma irradiation. Also BPV was effectively inactivated by 25 kGy of gamma irradiation. The cumulative log reduction factors of BHV, BVDV, BPIV-3, and BPV were ${\geq}$13.30, ${\geq}$14.32, ${\geq}$15.22, and ${\geq}$7.57, respectively. These results indicate that the production process for acelluar BAM has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.

Culture Conditions of Aspergillus oryzae in Dried Food-Waste and the Effects of Feeding the AO Ferments on Nutrients Availability in Chickens (건조한 남은 음식물을 이용한 Aspergillus oryzae균주 배양조건과 그 배양물 급여가 닭의 영양소 이용률에 미치는 영향)

  • Hwangbo J.;Hong E. C.;Lee B. S.;Bae H. D.;Kim W.;Nho W. G.;Kim J. H.;Kim I. H.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.291-300
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    • 2005
  • Two experiments were carried out to assess the appropriate incubation conditions namely; duration, moisture content and the ideal microbial inoculant for fermented dried food waste(EW) offered to broilers. The nutrient utilization of birds fed the FW diets at varying dietary inclusion rates was also compared with a control diet. In Experiment 1, different moisture contents(MC) of 30, 40, 50 and $60\%$ respectively were predetermined to establish the ideal duration of incubation and the microbial inoculant. A 1mL Aspergillus oryzae(AO) $(1.33\times10^5\;CFU/mL)$ was used as the seed inoculant in FW. This results indicated that the ideal MC for incubation was $40\~50\%$ while the normal incubation time was > 72 hours. Consequently, AO seeds at 0.25, 0.50, 0.75 and 1.00mL were inoculated in FW to determine its effect on AO count. The comparative AO count of FW incubated for 12 and 96 hours, respectively showed no significant differences among varying inoculant dosage rates. The FW inoculated with lower AO seeds at 0.10, 0.05 and 0.01mL were likewise incubated for 72 and 96 hours, respectively and no changes in AO count was detected(p<0.05). The above findings indicated that the incubation requirements for FW should be $%40\~50\%$ for 72 hours with an AO seed incoulant dosage rate of 0.10mL. Consequently, in Experiment II, after determining the appropriate processing condition for the FW, 20 five-week old male Hubbard strain were used in a digestibility experiment. The birds were divided into 4 groups with 5 pens(1 bird per pen). The dietary treatments were; Treatment 1 : Control(Basal diet), Treatment 2 : $60\%$ Basal+4$40\%$ FW, Treatment 3 : $60\%$ $Basal+20\%\;FW+20\%$ AFW(Aspergillus oryzae inoculate dried food-waste diet) and Treatment 4: $60\%$ Basal+$40\%$ Am. Digestibility of treatment 2 was lowed on common nutrients and amino acids compared with control(p<0.05) and on crude fat and phosphorus compared with AFW treatments(T3, T4)(plt;0.05). Digestibility of treatment 3 and 4 increased on crude fiber and crude ash compared treatment 2 (p<0.05). Digestibility of control was high on agrinine, leucine, and phenylalnine of essential amino acids compared with treatment 3 and 4(p<0.05), and diestibility of treatment 3 and 4 was improved on arginine, lysine, and threonine of essential amino acids. Finally, despite comparable nutrient utilization among treatments, birds fed the dietary treatment containing AO tended to superior nutrient digestion to those fed the $60\%$ Basa1+$40\%$ FW.

Studies on the Production of Alcohol from Woods (목재(木材)를 이용(利用)한 Alcohol 생산(生産)에 관(關)한 연구(硏究))

  • Cheong, Jin Cheol
    • Journal of Korean Society of Forest Science
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    • v.59 no.1
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    • pp.67-91
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    • 1983
  • In order to examine the alcohol production from softwoods (Pinus densiflora Sieb. et Zucc., Pinus rigida Miller, Larix leptolepis Gordon) and hardwoods (Alnus japonica Steud., Castanea crenata Sieb. et Zucc. Populus euramericana CV 214), chemical compositions were analyzed and conditions of acid hydrolysis with wood meals were established. Also strains which could remarkably decompose the cellulose were identified, and conditions of cellulase production of strains, characteristics of cellulase, and alcohol fermentation were examined. The results were summarized as follows. 1) In acid hydrolysis of wood, the high yield of reducing sugars was shown from 1.0% to 2.0% of hydrochloric acid and 2.0% of sulfuric acid. The highest yield was produced 23.4% at wood meals of Alnus japonica treated with 1.0% of hydrochloric acid. 2) The effect of raising the hydrolysis was good at $1.5kg/cm^2$, 30 times (acid/wood meal), and 45 min in treating hydrochloric acid and 30 min in treating sulfuric acid. 3) The pretreatments with concentrated sulfuric acid were more effective concentration ranged from 50% to 60% than that with hydrochloric acid and its concentration ranged from 50% to 60%. 4) The quantative analysis of sugar composition of acid hydrolysates revealed that glucose and arabinose were assayed 137.78mg and 68.24mg with Pinus densiflora, and 102.22mg and 65.89mg with Alnus janonica, respectively. Also xylose and galactose were derived. 5) The two strains of yeast which showed remarkably high alcohol productivity were Saccharomyces cerevisiae JAFM 101 and Sacch. cerevisiae var. ellipsoldeus JAFM 125. 6) The production of alcohol and the growth of yeasts were effective with the neutralization of acid hydrolysates by $CaCO_3$ and NaOH. Production of alcohol was excellent in being fermented between pH 4.5-5.5 at $30^{\circ}C$ and growth of yeasts between pH 5.0-6.0 at $24^{\circ}C$. 7) The production of alcohol was effective with the addition of 0.02% $(NH_2)_2CO$ and $(NH_4)_2SO_4$, 0.1% $KH_2PO_4$, 0.05% $MgSO_4$, 0.025% $CaCl_2$, 0.02% $MnCl_2$. Growth of yeasts was effective with 0.04-0.06% $(NH_2)_2CO$ and $(NH_4)_2SO_4$, 0.2% $K_2HPO_4$ and $K_3PO_4$, 0.05% $MgSO_4$, 0.025% $CaCl_2$, and 0.002% NaCl. 8) Among various vitamins, the production of alcohol was effective with the addition to pyridoxine and riboflavin, and the growth of yeasts with the addition to thiamin, Ca-pantothenate, and biotin. The production of aocohol was increased in 0.1% concentration of tannin and furfural, but mas decreased in above concentration. 9) In 100ml of fermented solution, alcohol and yeast were produced 2.201-2.275ml and 84-114mg for wood meals of Pinus densiflora, and 2.075-2.125ml and 104-128mg for that of Alnus japonica. Residual sugars were 0.55-0.60g and 0.60-0.65g for wood meals of Pinus densiflora and Alnus japonica, respectively, and pH varied from 3.3 to 3.6. 10) A strain of Trichoderma viride JJK. 107 was selected and identified as its having the highest activity of decomposing cellulose. 11) The highest cellulase production was good when CMCase incubated for 5 days at pH 6.0, $30^{\circ}C$ and xylanase at pH 5.0, $35^{\circ}C$. The optimum conditions of cellulase activity were proper in case of CMCase at pH 4.5, $50^{\circ}C$ and xylanase at pH 4.5, $40^{\circ}C$. 12) In fermentation with enzymatic hydrolysates, the peracetic acid treatment for delignification showed the best yields of alcohol and its ratio was effective with the addition of about 10 times. 13) The production of alcohol was excellent when wood meals and Koji of wheat bran was mixed with 10 to 8 and the 10g of wood meals of Pinus densiflora produced 2.01-2.14ml of alcohol and Alnus japonica 2.11-2.20ml.

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Oxidized LDL is a Chemoattractant for the Eosinophils and Neutrophils (산화 저비중 리포 단백이 호산구와 호중구에 대한 화학주성)

  • Hwang, Young-Sil;Lee, Jong-Deog;Busse, William B.
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.3
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    • pp.211-223
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    • 2001
  • Background : Rhinovirus infection of the airways results in increased permeability of the airway vascular endothelium with the influx of plasma proteins, including lipids such as LDL. In vitro studies on the effect of oxLDL on leukocytes has shown many pro inflammatory effects on multiple leukocytes. We hypothesized that oxLDL is one mechanism for recruiting granulocytes to the airways during a RV infection. Therefore, chemotaxis and transendothelial migration, in response to nLDL, was determined for these granulocytes. Methods : nLDL was oxidized with 5mM Cu2S04 for 20-24 hours. 3-5 105 cells were loaded into the Transwell filter while the chemotatic agonists were placed in the lower well for chemotaxis. Confluent monolayers on HPMEC were grown on Transwell filters for transendothelial migration. The filters were washed and eosinophils and neutrophils loaded on to the filter with the chemotatic agonist was were placed in the lower well. The wells were incubated for 3 hours. The number of migrating cells was counted on a hemocytometer. Results : OxLDL, but not nLDL, is chemotatic for eosinophils and neutrophils. The level of granulocytes chemotaxis was dependent on both the concentration of LDL and its degree of oxidation. OxLDL stimulates eosinophil and neutrophils migration across HPMEC monolayers (+/-IL-$1{\beta}$ preactivation) in a dose dependent manner. Conclusion : Increased vascular permeability during a RV infection may lead to the influx and oxidation of LDL. The resulting oxLDL. is one possible mechanism for the recruitment of neutrophils and eosinophils to the airway interstitial matrix. Once in the airways, granulocytes can further interact with oxLDL to promote airway inflammation.

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