• Title/Summary/Keyword: cell degradation

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Oxidative Pathway of $C^{14}-glucose$ in Various Human Cancer Tissues (각종 인체 암조직의 당의 산화경로 분석)

  • Lee, Bong-Kee;Lee, Sang-Don
    • The Korean Journal of Physiology
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    • v.2 no.1
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    • pp.23-30
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    • 1968
  • Tissue homogenates of 12 kinds of human cancer tissues were incubated separately in medium containing $C^{14}-1-glucose$ and $C^{14}-6-glucose$ as a substrate in order to observe the oxidative pathway of glucose in the tumor tissues. At the end of 3 hours incubation in the Dubnuff metabolic shaking incubator, respiratory $CO_2$ samples trapped by alkaling which was placed in the center well of incubation flask were analysed for total $CO_2$ production rates and their radioactivities. The tissue homogenate samples after incubation were analyzed for their concentrations of glucose, lactate and pyruvate. Calculations were made on the glucose consumption rate and accumulation rates of lactate and pyruvate. Fractionation of oxidative pathway of glucose was carried out by calculating $C^{14}O_2 yields from C-1 and C-6 carbon of glucose. The following results were obtained. 1. In 12 kinds of human cancer, total $CO_2$ production rates were less than $8{\mu}M/gm$ except 2 cases. These lower values impressed that oxidative metabolism in the tumor tissues generally inhibited as compared with that in normal tissues. On the other hand, fractions of $CO_2$ derived from glucose to total $CO_2$ production rates (RSA) were less than 10% in every case. These facts showed that oxidation of glucose into $CO_2$ was remarkably inhibited in the tumor tissues. 2. Factions of glucose disappeared into $CO_2\;(RGD_{CO_2})$, lactate $(RGD_L)$, pyruvate $(RGD_P)$ to glucose consumption rates were as follows. $RGD_{CO_2}$ were less than 2% in cases of in this experiment and $RGD_L$ showed more than 5% except in 2 cases. These facts showed that anaerobic degradation of glucose into 3 carbon compounds was easily proceeded but further degradation into $CO_2$ via the TCA cycle was greatly inhibited resulting in accumulation of lactate. There are large variation in values of $RGD_P$ in different kinds of tumor tissue but relatively higher values in $RGD_{CO_2}$ were obtained in the tumor tissues as compared with those of normal tissues. 3. The oxidative pathway of glucose in tumor tissues were analyzed from the values of RSA which were obtained in $C^{14}-1\;and\;C^{14}-6-glucose$ incubation experiments. It was found that 3% of $CO_2$ derived from glucose were oxidized via the principal EMP-TCA cycle and the remainder were via alternate pathway such as HMP in the liver cancer and values in other cancer tissues were as follows; 4% in the tongue cancer, 6% in the colon cancer, 6% in the lung cancer, 9% in the stomach cancer, 11% in the ovarian cancer, 12% in the neck tumor, 22% in the uterine cancer, 22% in the bladder tumor, 32% in the spindle cell sarcoma and 65% in the brain tumor. These values except later 2 cases showed less than 30% which is the lowest value among the normal tissues. Even in the brain tumor in which showed highest value in the tumor group. It is reasonable to suppose that this fraction was remarkably decreased because values in normal brain tissue was more than 90%. From the above data, it was concluded that in tumor tissues, oxidation of glucose via TCA cycle was greatly inhibited but correlation between degree of inhibited oxidation of glucose via TCA cycle and malignancy of tumor were not clarified in this experiments.

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Degradation of Poultry Feathers by Bacillus amyloliquefaciens Y10 With Plant Growth-promoting Activity and Biological Activity of Feather Hydrolyzates (식물 성장 촉진 활성을 가진 Bacillus amyloliquefaciens Y10에 의한 가금 우모의 분해 및 생산된 우모 분해산물의 생리활성)

  • Yedam Kim;Young Seok Lee;Youngsuk Kim;Jinmyeong Song;Yeongbeen Bak;Gyulim Park;O-Mi Lee;Hong-Joo Son
    • Journal of Life Science
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    • v.34 no.5
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    • pp.304-312
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    • 2024
  • This study was conducted to characterize strain Y10, isolated from discarded chicken feathers. Strain Y10 was identified as Bacillus amyloliquefaciens through phenotypic and 16S rRNA gene analysis. B. amyloliquefaciens Y10 exhibited plant growth-promoting activities, including the production of fungal cell-degrading enzymes (cellulase, lipase, protease, and pectinase), siderophores, ammonia, and indoleacetic acid. Furthermore, strain Y10 was able to inhibit the mycelial growth of several phytopathogenic fungi. When 0.1% sucrose as a carbon source and 0.05% casein as a nitrogen source were added to the basal medium, adjusted to pH 10, and cultured at 35℃, the degradation rate of chicken feathers by strain Y10 was about two times higher than that of the basal medium, with the feathers almost completely degraded in four days. Strain Y10 also degraded various keratin substrates, including duck feathers, wool, and human nails. It was confirmed that the feather hydrolyzates prepared using strain Y10 exhibited antioxidant activities, such as 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity (EC50 = 0.38 mg/ml) and superoxide dismutase-like activity (EC50 = 183.7 mg/ml). These results suggest that B. amyloliquefaciens Y10 is a potential candidate for the development of bioinoculants and feed additives applicable to the agricultural and livestock industries, as well as the microbiological treatment of keratin waste.

Effects of a Tetramethoxyhydroxyflavone on the Expression of Inflammatory Mediators in LPS-Treated Human Synovial Fibroblast and Macrophage Cells

  • Yoon, Do-Young;Cho, Min-Chul;Kim, Jung-Hee;Kim, Eun-Jin;Kang, Jeong-Woo;Seo, Eun-Hee;Shim, Jung-Hyun;Kim, Soo-Hyun;Lee, Hee-Gu;Oh, Goo-Taeg;Hong, Jin-Tae;Park, Joo-Won;Kim, Jong-Wan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.686-694
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    • 2008
  • The inhibitory effects of 5,6,3',5'-tetramethoxy 7,4'-hydroxyflavone (labeled as p7F) were elucidated on the productions of proinflammatory cytokines as well as inflammatory mediators in human synovial fibroblasts and macrophage cells. p7F inhibited IL-1${\beta}$ or TNF-${\alpha}$ induced expressions of inflammatory mediators (ICAM-1, COX-2, and iNOS). p7F also inhibited LPS-induced productions of nitric oxide and prostaglandin $E_2$ in RAW 264.7 cells. In order to investigate whether p7F would inhibit IL-1 signaling, p7F was added to the D10S Th2 cell line (which is responsive to only IL-1${\beta}$ and thus proliferates), revealing that p7F inhibited IL-1${\beta}$-induced proliferation of D10S Th2 cells in a dose-response manner. A flow cytometric analysis revealed that p7F reduced the intracellular level of free radical oxygen species in RAW 264.7 cells treated with hydrogen peroxide. p7F inhibited IkB degradation and NF-${\kappa}$B activation in macrophage cells treated with LPS, supporting that p7F could inhibit signaling mediated via toll-like receptor. Taken together, p7F has inhibitory effects on LPS-induced productions of inflammatory mediators on human synovial fibroblasts and macrophage cells and thus has the potential to be an anti-inflammatory agent for inhibiting inflammatory responses.

Change of Antioxidative Activity at Different Harvest Time and Improvement of Atopic Dermatitis Effects for Persimmon Leaf Extract (감잎(柿葉) 추출물의 채취시기에 따른 항산화 활성 변화 및 아토피성 피부염 개선효과)

  • Jung, Woo-Young;Jeong, Jong-Moon
    • The Korea Journal of Herbology
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    • v.27 no.1
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    • pp.41-49
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    • 2012
  • Objectives : The purpose of this study was to investigate the anti-oxidative and anti-atopic dermatitis effects of persimmon leaf extract obtained from Cheongdo-gun, where more than 60% of Korean persimmon is produced. Methods : Anti-oxidative effects of the crude persimmon leaf extract harvested monthly between May and November were determined by in vitro assay using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide dismutase (SOD)-like reaction. Anti-atopic dermatitis effects of the crude persimmon leaf extract were determined by using collagenase type I inhibition assay and by quantitative assays including serum histamine, prostaglandin E metabolite and leukotriene $B_4$ levels in animal model of atopic dermatitis using Balb/c mice. Results : Persimmon leaf extract harvested in May had higher levels of total phenolic compounds (182.24 mg/g) and flavonoids (23.05 mg/g) than the ones of different month extract. Also, persimmon leaf extract harvested in May showed the most effective extract scavenging activities of DPPH free radical ($13.39{\pm}0.21\;{\mu}g/ml$) and superoxide anion radical ($40.52{\pm}2.32\;{\mu}g/ml$), leading to use the persimmon leaf extract harvested in May for the experiments hereafter. Persimmon leaf extract showed $326.71{\pm}4.6\;{\mu}g/ml$ of 50% inhibitory concentration ($IC_{50}$) for collagenase type I which is responsible for the degradation of extracellular matrix. In addition, persimmon leaf extract application group could decrease serum levels of histamine, prostaglandin E metabolite and leukotriene $B_4$ compared to the negative control in animal model of atopic dermatitis. Especially, persimmon leaf extract showed a significantly decreased serum leukotriene $B_4$ level relative to the levels of histamine and prostaglandin E metabolite. Conclusions : Persimmon leaf extract showed anti-oxidative and anti-atopic dermatitis effects in vitro and in vivo. These results suggest that persimmon leaf extract may have immunoregulatory function for alleviating atopic dermatitis by decreasing collagenase activity and mast cell activation.

Identification of a Newly Isolated Protease-producing Bacterium, Bacillus subtilis FBL-1, from Soil (토양으로부터 새로이 분리된 단백질 분해효소 생산 미생물 Bacillus subtilis FBL-1의 동정)

  • Kim, Mina;Si, Jin-Beom;Wee, Young-Jung
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.185-193
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    • 2016
  • A novel proteolytic bacterium was isolated from soil at Yeungnam University, South Korea. The strain, named FBL-1, was rod-shaped with a smooth surface. Biolog and API 50CHB test results revealed that strain FBL-1 was a Bacillus species. Based on 16S rDNA sequencing and chemotaxonomic characterization, the strain was identified as Bacillus subtilis because it had the highest homology with Bacillus subtilis subsp. subtilis NCIB 3610 (99.5%). In liquid culture at 37℃ with shaking at 200 rpm, fructose and yeast extract were found to be the best carbon and nitrogen sources, respectively, for cell growth and protease production. The highest protease activity (451.640 U/ml) was obtained when the strain was cultured in medium containing 20 g/l of fructose and 5 g/l of yeast extract. Although further studies are needed to characterize the protease and enhance its activity, the newly isolated protein-degrading B. subtilis FBL-1 can be applicable for the production of peptides and for the degradation of proteins in various industries.

Behavior of Soluble Microbial Products in a Submerged Membrane Separation Activated Sludge Process (침지형 막분리 활성오니법에 있어서 생물대사성분의 거동)

  • Cha, Gi-Cheol;Lee, Dong-Yeol;Shim, Jin-Kie;Lee, Yong-Moo;Yoo, Ik-Keun;Ann, Seung-Ho
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.5
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    • pp.959-970
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    • 2000
  • A laboratory-scale experiment was conducted to investigate the effect of soluble microbial products(SMP) on permeate flux in the submerged membrane separation activated sludge process. Continuous and batch filtration test were operated to understand mechanism of relationship between membrane fouling and SMP. Synthetic wastewater(phenol) was used as a carbon source. Hydraulic retention time(HRT) and mixed-liquor volatile suspended solids(MLVSS) of the reactor were kept at 12 hours and 9.000mg VSS/L, respectively. Batch filtration tests ($J_{60}/J_o$) using the mixed liquor from reactor showed that the increase of accumulated SMP concentration in the reactor caused to the decreasing permeate flux and the increasing of the adhesion matters which form cake and gel layer. The resistance value of cake layer was measured $2.9{\times}10^{10}{\sim}4.0{\times}10^{10}(1/m)$, this value showed more significant effect on flux drop than that of among other resistance layers. Batch phenol-degradation experiment was conducted to observe SMP type expected $SMP_{nd}$ and $SMP_{e}$ (SMP resulted from endogenous cell decomposition), these are non-biodegradable high molecular weight organic matter and playa significant role in permeate flux drop. Also, SMP concentration was accumulated as increased of HRT against flux drop.

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The physiological and degradational characteristics of Fomers fomentarius (말굽버섯균의 생리적 특성과 부후특성)

  • Kim, Yu-I;Chai, Jyung-Ki
    • Journal of Mushroom
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    • v.2 no.4
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    • pp.200-206
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    • 2004
  • The optimal temperature for mycelial growth the F. fomentarius was $30^{\circ}C$ and the range of the temperature for mycelial growth wsa about 10~30. The optimal pH for the growth was 4.0. The percentage of weight loss percentage wsa 17.4%. The percentage of WEC extractives wsa increased to 2.24%. The observation of micromorphological showed that the detected cell wall were erosive and thinning as typical degradation pattern of white-rot fungi.

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Sanguinarine Induces Apoptosis in Human Hepatocellular Carcinoma HepG2 Cells through the Generation of ROS and Modulation of Akt/ERK Signaling Pathways (HepG2 인체 간암세포의 ROS 생성 및 ERK/Akt 신호전달 경로 조절을 통한 sanguinarine의 apoptosis 유도)

  • Hwang, Ju Yeong;Cho, Yung Hyun
    • Journal of Life Science
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    • v.25 no.9
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    • pp.984-992
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    • 2015
  • Sanguinarine is a benzophenanthridine alkaloid originally isolated from the roots of Sanguinaria canadensis. It has multiple biological activities (e.g., antioxidant and antiproliferative) and immune-enhancing potential. In this study, we explored the proapoptotic properties and modes of action of sanguinarine in human hepatocellular carcinoma HepG2 cells. Our results revealed that sanguinarine inhibited HepG2 cell growth and induced apoptosis in a dose-dependent manner. The induction of apoptosis by sanguinarine was associated with the up-regulation of Fas and Bax, the release of cytochrome c from the mitochondria to the cytosol, and the loss of the mitochondrial membrane potential. In addition, sanguinarine activated caspase-9 and -8, initiator caspases of the intrinsic and death extrinsic pathways, respectively, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose) polymerase. Sanguinarine also triggered the generation of reactive oxygen species (ROS). The elimination of ROS by N-acetylcysteine reversed sanguinarine-induced apoptosis. Furthermore, sanguinarine induced the dephosphorylation of Akt and the phosphorylation of mitogen-activated protein kinases, including extracellular signal-regulated kinase (ERK), c-jun N-terminal kinase (JNK), and p38. The growth inhibition was enhanced by the combined treatment of sanguinarine with a phosphatidylinositol 3'-kinase (PI3K) inhibitor and an ERK inhibitor but not JNK and p38 inhibitors. Overall, our data indicate that the proapoptotic effects of sanguinarine in HepG2 cells depend on ROS production and the activation of both intrinsic and extrinsic signaling pathways, which is mediated by blocking PI3K/Akt and activating the ERK pathway. Thus, our data suggest that sanguinarine may be a natural compound with potential for use as an antitumor agent in liver cancer.

Apoptotic Effect of Rubia cordifolia Dichloromethane Extracts on Human Acute Jurkat T Cells (천초근 dichloromethane 추출물의 Jurkat T 세포에서 세포사멸 효과)

  • Kim, Ji-Hye;Lee, Jong-Hwan;Kim, Young-Ho;Kim, Kwang-Hyeon
    • Journal of Life Science
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    • v.19 no.2
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    • pp.163-168
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    • 2009
  • To understand cytotoxic activity of Rubia cordifolia L. (Rubiaceae), which has been used as a traditional oriental medicine, the mechanism underlying cytotoxic effect of its extract on human acute Jurkat T cells was investigated. The methanol extract of roots (3 kg) of R. codifolia was evaporated, dissolved in water, and then extracted by dichloromethane. The substances in the chloroform extract showing the most cytotoxic activity were further purified by a series of preparative HPLC. The extracted active substance (65 mg) was designated as CCH1. When Jurkat T cells were treated with CCH1 at concentration ranging from 0.5 to 2.0 ${\mu}g$/ml, apoptotic phenomena of cells companying several subsequent biochemical reactions such as mitochondria cytochrome c release, activation of casapase-8, -9, and caspase- 3, degradation of PARP and DNA fragmentation occurred via mitochondria-dependent pathway. However, abrogation of apoptosis was observed in an ectopic expression of Bcl-xL, which is a suppressor for mitochondrial cytochrome c release. These results demonstrate that the cytotoxicity of CCH1 against Jurkat T cells is attributable to apoptosis mediated by mitochodria-dependent death-signaling regulated by Bcl-xL. In addition, the CCH1 is more potent to leukemia Jurkat T cell than to human peripheral blood monocyte cells (PBMC).

Absorbable Guided Bone Regeneration Membrane Fabricated from Dehydrothermal Treated Porcine Collagen (Dehydrothermal Treatment로 제작한 흡수성 콜라겐 골유도재생술 차단막)

  • Pang, Kang-Mi;Choung, Han-Wool;Kim, Sung-Po;Yang, Eun-Kyung;Kim, Ki-Ho;Kim, Soung-Min;Kim, Myung-Jin;Jahng, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.33 no.2
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    • pp.112-119
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    • 2011
  • Purpose: Collagen membranes are used extensively as bioabsorbable barriers in guided bone regeneration. However, collagen has different effects on tissue restoration depending on the type, structure, degree of cross-linking and chemical treatment. The purpose of this study was to evaluate the inflammatory reaction, bone formation, and degradation of dehydrothermal treated porcine type I atelocollagen (CollaGuide$^{(R)}$) compared to of the non-crosslinked porcine type I, III collagen (BioGide$^{(R)}$) and the glutaldehyde cross-linked bovine type I collagen (BioMend$^{(R)}$) in surgically created bone defects in rat mandible. Methods: Bone defect model was based upon 3 mm sized full-thickness transcortical bone defects in the mandibular ramus of Sprague-Dawley rats. The defects were covered bucolingually with CollaGuide$^{(R)}$, BioMend$^{(R)}$, or BioGide$^{(R)}$ (n=12). For control, the defects were not covered by any membrane. Lymphocyte, multinucleated giant cell infiltration, bone formation over the defect area and membrane absorption were evaluated at 4 weeks postimplantation. For comparison of the membrane effect over the bone augmentation, rats received a bone graft plus different covering of membrane. A $3{\times}4$ mm sized block graft was harvested from the mandibular angle and was laid and stabilized with a microscrew on the naturally existing curvature of mandibular inferior border. After 10 weeks postimplantation, same histologic analysis were done. Results: In the defect model at 4 weeks post-implantation, the amount of new bone formed in defects was similar for all types of membrane. Bio-Gide$^{(R)}$ membranes induced significantly greater inflammatory response and membrane resorption than other two membranes; characterized by lymphocytes and multinucleated giant cells. At 10 weeks postoperatively, all membranes were completely resorbed. Conclusion: Dehydrotheramal treated cross-linked collagen was safe and effective in guiding bone regeneration in alveolar ridge defects and bone augmentation in rats, similar to BioGide$^{(R)}$ and BioMend$^{(R)}$, thus, could be clinically useful.