• Title/Summary/Keyword: CHO-K1 cell

Search Result 2,622, Processing Time 0.033 seconds

Effect of $H_2O_2$ on Alveolar Epithelial Barrier Properties (폐상피세포 장벽에 대한 $H_2O_2$의 영향)

  • Suh, Duk-Joon;Cho, Se-Heon;Kang, Chang-Woon
    • Tuberculosis and Respiratory Diseases
    • /
    • v.40 no.3
    • /
    • pp.236-249
    • /
    • 1993
  • Background: Among the injurious agents to which the lung airspaces are constantly exposed are reactive species of oxygen. It has been widely believed that reactive oxygen species may be implicated in the etiology of lung injuries. In order to elucidated how this oxidant causes lung cell injury, we investigated the effects of exogenous $H_2O_2$ on alveolar epithelial barrier characteristics. Methods: Rat type II alveolar epithelial cells were plated onto tissue culture-treated polycarbonate membrane filters. The resulting confluent monolayers on days 3 and 4 were mounted in a modified Ussing chamber and bathed on both sides with HEPES-buffered Ringer solution. The changes in short-circuit current (Isc) and monolayer resistance (R) in response to the exogenous hydroperoxide were measured. To determine the degree of cellular catalase participation in protection against $H_2O_2$ injury to the barrier, experiments were repeated in the presence of 20 mM aminotriazole (ATAZ, an inhibitor of catalase) in the same bathing fluid as the hydroperoxide. Results: These monolayers have a high transepithelial resistance (>2000 ohm-$cm^2$) and actively transport $Na^+$ from apical fluid. $H_2O_2$(0-100 mM) was then delivered to either apical or basolateral fluid. Resulting indicated that $H_2O_2$ decreased Isc and R gradually in dose-dependent manner. The effective concentration of apical $H_2O_2$ at which Isc (or R) was decreased by 50% at one hour ($ED_{50}$) was about 4 mM. However, basolateral $H_2O_2$ exposure led to $ED_{50}$ for Isc (and R) of about 0.04 mM. Inhibition of cellular catalase yielded $ED_{50}$ for Isc (and R) of about 0.4 mM when $H_2O_2$ was given apically, while $ED_{50}$ for basolateral exposure to $H_2O_2$ did not change in the presence of ATAZ. The rate of $H_2O_2$ consumption in apical and basolateral bathing fluids was the same, while cellualr catalase activity rose gradually with time in culture. Conclusion: Our data suggest that basolateral $H_2O_2$ may affect directly membrane component (e.g., $Na^+,\;K^+$-ATPase) located on the basolateral cell surface. Apical $H_2O_2$, on the other hand, may be largely degraded by catalase as it passes through the cells before reaching these membrane components. We conclude that alveolar epithelial barrier integrity as measured by Isc and R are compromised by $H_2O_2$ being relatively sensitive to basolateral (and insensitive to apical) $H_2O_2$.

  • PDF

The Increase of Apoptotic Neutrophils and Phagocytic Macrophage by Germanium in Acute Lung Injury Induced by Lipopolysaccharide (LPS에 의한 급성 폐손상에서 게르마늄에 의한 호중구 세포사와 큰포식세포의 포식능 증가)

  • Lee, Yoon-Jeong;Cho, Hyun-Gug;Jeune, Kyung-Hee
    • Applied Microscopy
    • /
    • v.38 no.4
    • /
    • pp.293-306
    • /
    • 2008
  • The essential factor of acute respiratory distress syndrome (ARDS), an acute lung injury accompanied commonly by sepsis syndrome is accumulation of neutrophils in lung tissue. The study attempted to confirm whether a lung injury would be decreased with the anti-inflammatory effect of germanium by the treated germanium prior to the development of ARDS and whether nitric oxide influence in suppressing a lung injury. Test groups were divided in the following structure for experiment; CON that has been administered with sodium chloride to airway, LPS administered with endotoxin for 5 hours in the same amount and 5 hours of endotoxin administered Ge+LPS following 1 hours of pre-treated germanium. The result of a test using experimental animals, infilteration of neutrophils (p<0.001) in bronchoalveolar lavage fluid (BALF) was significantly decreased, the structure of lung tissue was preserved relatively well, and much neutrophils with distinct positive were observed on tunel staining which showed increase of apoptotic neutrophils in the pre-treated germanium group compare to the endotoxin administrated group. In observation of ultrastructural changes of cell in BALF, phagocytic alveolar macrophage was increased in alveolar space, the nucleus of most engulfed neutrophils were condensed, and some apoptosis neutrophils appears to be DNA fragmentation and effacement of cellular organelles were found in intercellular matrix in the pre-treated germanium group. However, the nitric oxide showed increase in all the groups excluding CON, and the nitric oxide effect such as degranulation diminishing of mast cells and apoptosis increase of neutrophils in the pre-treated group only. The situation appears that there was change in internal environment of the experimental animal by the pre-treated germanium before the nitric oxide is produced and the anti-inflammatory effect activated the pre-processed germanium by nitric oxide which activated following the change. Therefore, the nitric oxide created from macrophage in accordance with the pre-treated germanium appears to influence in alleviating a lung injury. Accordingly, acute lung injury is alleviated by the anti-inflammatory effect of germanium such as inhibition of neutrophils migration, induction of neutrophil apoptosis and increase of phagocytic function of phagocyte, and the nitric oxide produced from activated macrophage by germanium would influence in suppressing a lung injury.

Changes in Quality Characteristics of Seasoned Soy Sauce Treated with Superheated Steam and High Hydrostatic Pressure during Cold Storage (과열증기와 초고압 처리법을 적용한 간장 소스의 냉장저장 중 품질 특성 변화)

  • Choi, Yoon;Oh, Ji-Hye;Bae, In-Young;Cho, Eun-Kyoung;Kwon, Dae-Joong;Park, Hae-Won;Yoon, Sun
    • Korean journal of food and cookery science
    • /
    • v.29 no.4
    • /
    • pp.387-398
    • /
    • 2013
  • Seasoned soy sauce is one of the popular seasoning sauces added to the Korean traditional foods such as Bulgogi, Galbi. However, industrially processed sauces have poor sensory quality because of heating treatment for sterilization. The purpose of this study was to develop seasoned soy sauce having fresh taste and good quality by applying superheated steam (SHS) and high hydrostatic pressure (HHP) technologies. To maintenance the sauce qualities, food materials such as apple, onion, and garlic were pretreated with SHS (heater $100^{\circ}C$, steam $280^{\circ}C$, 30 s~1 min 30 s) before mixing with other ingredients. During storage of 7 days, color, pH, and browning potential of SHS treated samples (apple, onion and garlic) did not change and also polyphenol oxidase was inactivated (p<0.05). The seasoned soy sauce including SHS treated materials was sterilized by thermal process ($85^{\circ}C$, 30min) or non-thermal process, HHP (550 MPa, $5{\sim}10^{\circ}C$, 3 min). In SHS+HHP treated sauce, salinity, sugar contents, lightness, viscosity did not change (p<0.05), and total viable cell counts were detected below 4 log cycle at $5^{\circ}C$ for 30 days. E.coli and B.cereus are not determined in all samples. In sensory evaluation, Bulgogi prepared with SHS+HHP treated sauce was more acceptable than others.

Results of MRI Evaluation for the Fatty Masses (지방 종괴의 진단에 대한 MRI의 판별 능력)

  • Seo, Jae-Sung;Ahn, Jong-Chul;Kim, Jeong-Rae;Choi, Jun-Hyuk;Cho, Kil-Ho;Shin, Duk-Seop
    • The Journal of the Korean bone and joint tumor society
    • /
    • v.11 no.1
    • /
    • pp.25-31
    • /
    • 2005
  • Purpose: This study was designed to know the usefulness of the MRI to distinguish lipoma and well differentiated liposarcoma (WDL). Materials and methods: 47 lipomatous tumors with MRI were reviewed among the 107 lipomatous tumors operated in our department. MRI examinations and their corresponding pathology reports were compared to determine sensitivity, specificity, diagnostic ability, positive predictable value and negative predictable value. Statistical analysis was performed to know the relationship between malignancy of the tumor (WDL) with the age and gender of the patients, and location, depth, size and the enhancement of tumors in MRI. Results: Among 28 lipoma in MRI examinations, 26 were proved as lipoma in pathology, and only 6 were WDL from 19 suspicious lesions in MRI, and others were proved as lipoma variants mostly. The varieties of lipoma variants were fibrolipoma, angiolipoma, spindle cell lipoma, lipoblastoma and angiomyolipoma. The sensitivity, specificity, diagnostic ability, positive predictable value and negative predictable value of MRI were 100%, 68 %, 72%, 31% and 100% in WDL, and 90%, 89%, 89%, 93% and 84% in lipoma. Among the variants to distinguish WDL and lipoma, the size of tumor and enhancement in MRI were significant statistically (p<0.05). Conclusion: MRI was highly sensitive in detection of WDL and highly specific in detection of simple lipoma. The size of tumor and enhancement in MRI were significant variants to distinguish WDL and lipoma. When MRI finding is non-specific, it is more likely to represent one of lipoma variants.

  • PDF

Physicochemical properties of spray-dried rice flour with Lactobacillus plantarum CGKW3 (분무건조공정을 이용한 유산균포집 미분의 제조 및 물리화학적 특성)

  • Park, Hye-Mi;Lee, Dae-Hoon;Jeong, Yoo-Seok;Jung, Hee-Kyoung;Cho, Jae-Gon;Hong, Joo-Heon
    • Food Science and Preservation
    • /
    • v.22 no.3
    • /
    • pp.392-398
    • /
    • 2015
  • The physicochemical properties of spray-dried rice flour with Lactobacillus plantarum CGKW3 were investigated. Amylose and damaged starch contents of spray-dried rice flour (S10, S20, S30, and S50) with L. plantarum CGKW3 were 14.18~17.75% and 24.65~34.08%, respectively. The particle size of spray-dried rice flour was $82.28{\sim}131.17{\mu}m$. The rice flour with L. plantarum CGKW3 showed a good powder flowability. The water absorption and water solubility of spray-dried rice flour were 1.96~2.13 and 9.91~21.95%, respectively. Thermal properties measured by differential scanning calorimeter revealed that the enthalpy (${\Delta}H$) for starch gelatinization were highest in the rice flour (S50) with L. plantarum CGKW3. When compared, the viable cell number of spray-dried rice flour were found to be in the following order: S10 (5.78 log CFU/g) < S20 (6.38 log CFU/g) < S30 (6.69 log CFU/g) < S50 (7.11 log CFU/g). The survaival rate of L. plantarum CGKW3 was 60.02-73.85%, which reflected the improvement in the quality of rice flour with an increase in treatment concentration. Based on our results, spray-dried rice flour with L. plantarum CGKW3 could be used in various types of rice foods.

Analysis of p53 and Retinoblasoma(Rb) Gene Polymorphisms in Relation to Lung Cancer in Koreans (한국인 폐암 환자에 대한 p53 및 Rb유전자의 다형성 분석)

  • Lee, Kyung-Sang;Sohn, Jang-Won;Yang, Suck-Chul;Yoon, Ho-Joo;Shin, Dong-Ho;Park, Sung-Soo;Lee, Jung-Hee;Lee, Chun-Geun;Cho, Youl-Hee
    • Tuberculosis and Respiratory Diseases
    • /
    • v.44 no.3
    • /
    • pp.534-546
    • /
    • 1997
  • Background : The p53 and retinoblastoma(Rb) tumor suppressor genes are associated with the pathogenesis of several types of human cancer. Substantial proportion of the primary lung cancers or cell lines have been reported to have the p53 and/or the Rb gene mutations. But, so far there is no report on the analysis of the Rb gene polymorphism as one of the genetic susceptibility marker. This study was undertaken to establish the gene frequencies of the polymorphic genotypes of the p53 and Rb genes in Koreans to evaluate the possible involvement of these genotypes as a risk factor of lung cancer. Methods : In this study 145 controls without previous and present tumor history and 128 lung cancer patients were subjected to analysis. The two intragenic polymorphisms of the p53 gene(exon 4/ AccII, intron 6/MspI) and one intron 17/XbaI polymorphism of the Rb gene were analysed by the method of polymersae chain reaction- restriction fragment length polymorphisms(PCR-RFLPs). The genotype of the intron 3/16 bp repeat polymorphism of p53 was determined by PCR and direct gel electrophoresis. Results : There were no significant differences in the genotype distributions of the p53 gene between lung cancer patients and controls. But heterozygotes(Arg/Pro) of the exon 4/AccII polymorphisms were slightly over-represented than controls, especially in the Kreyberg type I cancer, which was known to be associated with smoking. The intron 3/16 bp duplication and the intron 6/MspI polymorphisms were in complete linkage disequilibrium. About 95% of the individuals were homozygotes of the common alleles both in the 16 duplication and MspI polymorphisms, and no differences were deteced in the genotype distributions between lung cancer patients and controls. Overall genotype distributions of the Rb gene polymorphisms between lung cancer patients and controls were not significantly different However, the genotype distributions in the Kreyberg type I cancer were significantly different from those of controls(p = 0.0297) or adenocarcinomas(p = 0.0008). It was noticeable that 73.4% of the patients with adenocarcinomas were heterozygotes(r1/r2) whereas 39.2% of the Kreyberg type I cancer were heterozygous at this polymorphisms. In the lung cancer patients, significant differences were also noted between the high dose smokers and low dose smokers including non-smokers(p = 0.0258). The relative risk to Kreyberg type I cancer was significantly reduced in the individuals with the genotype of r1/r2(odds ratio = 0.46, 95% C.I. = 0.25-0.86, p = 0.0124). The combined genotype distribution of the exon 4 AccII of the p53 and the intron 17 Rb gene polymorphisms in Kreyberg type I cancers were significantly different from dose of controls or adenocarcinomas. The highest odds ratio were observed in the individuals with the genotypes of Arg/Pro and r2/r2(odds ratio = 1.97,95% C.I. = 0.84-4.59) and lowest one was in the patients with Arg/Arg, r1/r2 genotype(odds ratio = 0.54, 95% C.I. = 0.25-1.14). Conclusion : The p53 and the Rb gene polymorphisms modulate the risk of smoking induced lung cancer development in Koeans. However, the exact mechanism of risk modulation by these polymorphism remains to be determined. For more discrete clarification of associations between specific genotypes and lung cancer risk, the evaluations of these polymorphisms in other ethnics and more number of patients will be needed.

  • PDF

Effects of Linoleic Acid and Serum Albumin Concentrations on Lipid Metabolism in HepG2 Cells (간세포 배양에서 Linoleic Acid와 혈청알부민의 첩가가 지질대사에 미치는 영향)

  • Cha, Jae-Young;Cho, Young-Su
    • Applied Biological Chemistry
    • /
    • v.42 no.3
    • /
    • pp.229-234
    • /
    • 1999
  • The effects of linoleic acid(LA, 18 : 2) and/or bovine serum albumin(BSA) on the lipid metabolism in human hepatoma cell line HepG2 cells were evaluated. HepG2 cells were cultured in basal Dulbecco's modified Eagle's(DME) medium(Basal medium), DME medium containing 0.2 mM LA(LA medium), or DME medium containing both 0.2 mM LA and 0.2-1.0% BSA(LA+BSA medium). $[^{14}C]Acetate(0.3\;{\mu}Ci/ml\;medium)$ was added as a radioactive lipid precursor and the cells were incubated for 6 hours. An addition of LA to basal medium resulted in a decrease in the incorporation of $[^{14}C]acetate$ into total cholesterol fraction. In contrast, an addition of BSA to LA-containing medium tended to increase the incorporation of $[^{14}C]acetate$ into total cholesterol. The alteration of cholesterol metabolism in HepG2 cells incubated in LA+BSA medium was attributed by an increase in the incorporation of $[^{14}C]acetate$ into free cholesterol, but not cholesteryl ester fraction. In addition, the secretion of cholesterol was increased by LA+BSA medium, suggesting that BSA stimulates cholesterol secretion. No significant change in the incorporation of $[^{14}C]acetate$ into cellular total lipids was observed among the experimental groups. However, an increased incorporation of $[^{14}C]-labelled$ fatty acid into cellular triacylglycerol and decreased incorporation into phospholipid were observed in cells incubated with LA+BSA medium as compared to those of LA medium. The secretions of $[^{14}C]-labelled$ triacylglycerol, phospholipid, and free fatty acid were also stimulated in HepG2 cells incubated with LA+BSA medium. In conclusion, the present study suggests that in human hepatocytes, LA and BSA influence lipid metabolism, and BSA enhances the secretion of lipids.

  • PDF

Characterization of Microsomal $Ca^{2+}$ Uptake in Tomato Root Tissues (토마토 뿌리조직에서 분리한 마이크로솜의 $Ca^{2+}$ 흡수 특성)

  • Cho, Kwang-Hyun;Kim, Young-Kee
    • Applied Biological Chemistry
    • /
    • v.42 no.2
    • /
    • pp.116-122
    • /
    • 1999
  • In order to characterize the property of $Ca^{2+}$ transport in plant cell, microsomes were prepared from the roots of tomato and microsomal $^{45}Ca^{2+}$ uptake was measured. When 1 mM vanadate, a selective inhibitor of P-type ATPases, 50 mM $NO_3^-$, a specific inhibitor of vacuolar $H^{+}-ATPase$, and both of these inhibitors were treated, the microsomal $^{45}Ca^{2+}$ uptakes were inhibited by 20, 33 and 47%, respectively. The inhibitory effects of these two inhibitors were investigated by using a protonophore, gramicidin. When the chemical gradient of $H^{+}$ was relieved by gramicidin, the uptake was decreased by 30%, implying the presence of $Ca^{2+}/H^+$ antiporter in the microsomal membrane. In the $^{45}Ca^{2+}$ uptake experiment, the effect of gramicidin was independent of vanadate-induced inhibition. However, when the activity of vacuolar $H^{+}-ATPase$ was inhibited by $NO_3^-$, the effect of gramicidin was severely decreased. Meanwhile, thapsigargin, a specific antagonist of ER/SR-type $Ca^{2+}-ATPase$, inhibited the microsomal $^{45}Ca^{2+}$ uptake and the maximum inhibitory effect was obtained at $10\;{\mu}M$. The effect of thapsigargin was blocked by $NO_3^-$ and gramicidin, but not by vanadate. These results imply that vanadate directly inhibits the activity of $Ca^{2+}-ATPase$; however, $NO_3^-$ and thapsigargin block the activity of $Ca^{2+}/H^+$ antiporter by inhibiting the vacuolar $H^{+}-ATPase$. In conclusion, the microsomal $^{45}Ca^{2+}$ uptakes are mediated by two major enzymes, $Ca^{2+}-ATPase$ and $Ca^{2+}/H^+$ antiporter in tomato root tissue.

  • PDF

Tumor-suppressor Protein p53 Sensitizes Human Colorectal Carcinoma HCT116 Cells to 17α-estradiol-induced Apoptosis via Augmentation of Bak/Bax Activation (17α-Estradiol에 의한 인체 대장암 세포주 HCT116의 에폽토시스에 수반되는 Bak/Bax의 활성화에 미치는 종양억제단백질 p53의 강화효과)

  • Han, Cho Rong;Lee, Ji Young;Kim, Dongki;Kim, Hyo Young;Kim, Se Jin;Jang, Seokjoon;Kim, Yoon Hee;Jun, Do Youn;Kim, Young Ho
    • Journal of Life Science
    • /
    • v.23 no.10
    • /
    • pp.1230-1238
    • /
    • 2013
  • The regulatory effect of the tumor-suppressor protein p53 on the apoptogenic activity of $17{\alpha}$-estradiol ($17{\alpha}-E_2$) was compared between HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$) cells. When the HCT116 ($p53^{+/+}$) and HCT116 ($p53^{-/-}$) cells were treated with $2.5{\sim}10{\mu}M$ $17{\alpha}-E_2$ for 48 h or with $10{\mu}M$for various time periods, cytotoxicity and an apoptotic sub-$G_1$ peak were induced in the HCT116 ($p53^{+/+}$) cells in a dose- and time-dependent manner. However, the HCT116 ($p53^{-/-}$) cells were much less sensitive to the apoptotic effect of $17{\alpha}-E_2$. Although $17{\alpha}-E_2$ induced aberrant mitotic spindle organization and incomplete chromosome congregation at the equatorial plate, $G_2/M$ arrest was induced to a similar extent in both cell types. In addition, $17{\alpha}-E_2$-induced activation of Bak and Bax, ${\Delta}{\Psi}m$ loss, and PARP degradation were more dominant in the HCT116 ($p53^{+/+}$) than in the HCT116 ($p53^{-/-}$) cells. In accordance with enhancement of p53 phosphorylation (Ser-15) and p53 levels, p21 and Bax levels were elevated in the HCT116 ($p53^{+/+}$) cells treated with $17{\alpha}-E_2$. The HCT116 ($p53^{-/-}$) cells exhibited barely or undetectable levels of p21 and Bax, regardless of $17{\alpha}-E_2$ treatment. On the other hand, although the level of Bcl-2 was slightly lower in the HCT116 ($p53^{+/+}$) than in the HCT116 ($p53^{-/-}$) cells, it remained relatively constant after the $17{\alpha}-E_2$ treatment. Together, these results show that among the components of the $17{\alpha}-E_2$-induced apoptotic-signaling pathway, which proceeds through mitotic spindle defects causing mitotic arrest, subsequent activation of Bak and Bax and the mitochondria-dependent caspase cascade, leading to PARP degradation, $17{\alpha}-E_2$-induced activation of Bak and Bax is the upstream target of proapoptotic action of p53.

THE EFFECTS OF THE PLATELET-DERIVED GROWTH FACTOR-BB ON THE PERIODONTAL TISSUE REGENERATION OF THE FURCATION INVOLVEMENT OF DOGS (혈소판유래성장인자-BB가 성견 치근이개부병변의 조직재생에 미치는 효과)

  • Cho, Moo-Hyun;Park, Kwang-Beom;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
    • /
    • v.23 no.3
    • /
    • pp.535-563
    • /
    • 1993
  • New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration alre basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, biological mediators. Platelet-derived growth factor (PDGF) is one of polypeptide growth factor. PDGF have been reported as a biological mediator which regulate activities of wound healing progress including cell proliferation, migration, and metabolism. The purposes of this study is to evaluate the possibility of using the PDGF as a regeneration promoting agent for furcation involvement defect. Eight adult mongrel dogs were used in this experiment. The dogs were anesthetized with Pentobarbital Sodium (25-30 mg/kg of body weight, Tokyo chemical Co., Japan) and conventional periodontal prophylaxis were performed with ultrasonic scaler. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree III furcation defect was made on mandibular second(P2) and fourth(P4) premolar. For the basic treatment of root surface, fully saturated citric acid was applied on the exposed root surface for 3 minutes. On the right P4 20ug of human recombinant PDGF-BB dissolved in acetic acid was applied with polypropylene autopipette. On the left P2 and right P2 PDGF-BB was applied after insertion of ${\beta}-Tricalcium$ phosphate(TCP) and collagen (Collatape) respectively. Left mandibular P4 was used as control. Systemic antibiotics (Penicillin-G benzathine and penicillin-G procaine, 1 ml per 10-25 1bs body weight) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operated sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 2, 4, 8, 12 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with H-E staining. At 2 weeks after surgery, therer were rapid osteogenesis phenomenon on the defected area of the PDGF only treated group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. New cementum fromation was observed from 2 weeks after surgery, and the thickness was increased until 8 weeks with typical Sharpey’s fibers reembedded into new bone and cementum. In both PDGF-BB with TCP group and PDGF-BB with Collagen group, regeneration process including new bone and new cementum formation and the group especially in the early weeks. It might be thought that the migration of actively proliferating cells was prohibited by the graft materials. In conclusion, platelet-derived growth factor can promote rapid osteogenesis during early stage of periodontal tissue regeneration.

  • PDF