• Title/Summary/Keyword: prolonged culture

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Lifespan Extension Property of Quercetin-3-O-${\beta}$-D-glucopyranoside-7-O-${\alpha}$-L-rhamnopyranoside from Curcuma longa L. In Caenorhabditis elegans (강황지하부 부산물에서 분리한 Quercetin-3-O-${\beta}$-D-glucopyranoside-7-O-${\alpha}$-L-rhamnopyranoside가 선충의 수명연장에 미치는 영향)

  • Ahn, Dalrae;Lee, Eun Byeol;Kim, Ban Ji;Lee, So Yeon;Ahn, Min-Sil;Eun, Jae Soon;Shin, Tae-Yong;Kim, Dae Keun
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.275-281
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    • 2014
  • After harvesting the medicinal parts of Curcuma longa, the remaining underground parts were discarded. From the remaining underground parts of Curcuma longa quercetin-3-O-${\beta}$-D-glucopyranoside-7-O-${\alpha}$-L-rhamnopyranoside (Q37) was isolated. The antioxidant activities in vitro and lifespan-extension effect of Q37 were elucidated using the Caenorhabditis elegans. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging effect of Q37 showed similar potent activities in comparison with vitamin C. Q37 also showed potent superoxide quenching activities as measured by the riboflavin- and xanthine-originated superoxide quenching activity tests. Q37 prolonged lifespan of worms under normal culture condition. In terms of protective effect of Q37 on the stress conditions such as thermal and oxidative stresses, Q37-treated worms exhibited enhanced survival rate, as compared to control worms. To know the possible mechanism of Q37-mediated increased lifespan and stress resistance of worms, we examined the activities of Q37on superoxide dismutase (SOD), and invested intracellular reactive oxygen species (ROS) levels. The results revealed that Q37 was able to elevate SOD activity of worms and reduce intracellular ROS accumulation in a dose-dependent manner.

'HWABYUNG' in the view of Oriental Medinine (한의학에서 본 홧병의 해석)

  • Kim Jong-Woo;Whang Wei-Wan
    • Journal of Oriental Neuropsychiatry
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    • v.5 no.1
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    • pp.9-15
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    • 1994
  • Hwabyung is a culture-related disease generally know among people in Korea, and various studies have been done by the Psychiatrists and by the Oriental Medical Doctors. While the Psychiatrists explain the Hwabyung as a disease having a unique patholoical process, the Oriental Medical Doctors explain it as a symptoms having the character of fire. When interpreted in Oriental Medical point of view, this can be concluded as 1)the cause of the disease is the depression of liver-energy(肝氣鬱結) induced by emotional suppression, 2)the reason why the disease occurs mostly to female is that they usually fail overcoming stresses and have physical and visceral weaknesses, 3)the symptoms are associated with the dynamics of fire, because the disease originates from the rising character of fire and from the inadequit supression of extreme heart-fire(心火) by the kidney-wter(腎水), 4)the reason why the disease goes through chronic process is because of the complexed emotions, converted fire caused by prolonged emotional disorder and inapropreate supression of fire with grow older. This results suggest thst the cooperative study of the clinical examination of the Psychiatrsis and the study on the fire by the Oriental Medical Doctors is necessary to establish the process of the disease, the pathological procedure, the prognosis and the method of treatment.

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Clinical Efficacy of Radiation-Sterilized Allografts for Sellar Reconstruction after Transsphenoidal Surgery

  • Kim, Se-Jin;Jeon, Chi-Man;Kong, Doo-Sik;Park, Kwan;Kim, Jong-Hyun
    • Journal of Korean Neurosurgical Society
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    • v.50 no.6
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    • pp.503-506
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    • 2011
  • Objective : The aim of this study was to assess the safety and efficacy of radiation-sterilized allografts of iliac bone and fascia lata from cadaver specimens to repair skull base defects after transsphenoidal surgery. Methods : Between May 2009 and January 2010, 31 consecutive patients underwent endonasal transsphenoidal surgery and all patients received sellar reconstruction using allografts following tumor removal. The allografts were obtained from the local tissue bank and harvested from cadaver donors. The specimens used in our approach were tensor fascia lata and the flat area of iliac bone. For preparation, allografts were treated with gamma irradiation after routine screening by culture, and then stored at $-70^{\circ}C$. Results : The mean follow-up period after surgery was 12.6 months (range, 7.4-16 months). Overall, postoperative cerebrospinal fluid (CSF) leaks occurred in three patients (9.7%) and postoperative meningitis in one patient (3.2%). There was no definitive evidence of wound infection at the routine postoperative follow-up examination or during re-do surgery in three patients. Postoperative meningitis in one patient was improved with the use of antibiotics and prolonged CSF diversion. Conclusion : We suggest that allograft materials can be a feasible alternative to autologous tissue grafts for sellar reconstruction following transsphenoidal surgery under selected circumstances such as no or little intraoperative CSF leaks.

A High-Yielding, Generic Fed-Batch Process for Recombinant Antibody Production of GS-Engineered Cell Lines

  • Fan, Li;Zhao, Liang;Sun, Yating;Kou, Tianci;Zhou, Yan;Tan, Wen-Song
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1695-1702
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    • 2009
  • An animal-component-free and chemically defined fed-batch process for GS-engineered cell lines producing recombinant antibodies has been developed. The fed-batch process relied on supplying sufficient nutrients to match their consumption, simultaneously minimizing the accumulation of by-products (lactate and osmolality). The proportionalities of nutritional consumption were determined by direct analysis. The robust, metabolically responsive feeding strategy was based on the offline measurement of glucose. The fed-batch process was shown to perform equivalently in GS-CHO and GS-NS0 cultures. Compared with batch cultures, the fed-batch technology generated the greater increase in cell yields (5-fold) and final antibody concentrations (4-8-fold). The majority of the increase in final antibody concentration was a function of the increased cell density and the prolonged culture time. This generic and high-yielding fed-batch process would shorten development time, and ensure process stability, thereby facilitating the manufacture of therapeutic antibodies by GS-engineered cell lines.

Viability and Functions of Alginate-microencapsulated Islets Isolated from Neonatal Pigs

  • Lin, Yi-Juain;Wang, Jui-Ping;Chung, Yu-Tung;Sun, Yu-Ling;Chou, Yu-Chi
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.5
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    • pp.795-801
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    • 2007
  • Patients with Type I diabetes mellitus have been treated with porcine insulin for several decades and pigs have recently been deemed an ideal source of microencapsulated islet cells for clinical xenotransplantation. In this study, neonatal pigs were anesthetized and sacrificed prior to a pancreatectomy. Islet cells were isolated from pancreas via collagenase digestion. Islet cells were separated and collected by hand under microscopic guidance. These cells were suspended in 1.4% sodium alginate solution and encapsulated by dropping them into 1.1% calcium chloride solution and in which the round gel in size was 250-400 ${\mu}m$ in diameter. Viability of the microencapsulated islet cells cultured in medium at $37^{\circ}C$ was assessed by MTT assay. Furthermore, insulin released in response to glucose challenge was investigated using an enzyme-linked immunosorbent assay. Secretion of insulin was low in response to the basal glucose solution (4.4 mM) in medium and was significantly higher in response to the high glucose solution (16.7 mM). The viability of microencapsulated islet cells did not differ significantly over a period of 7 days; that is, the increasing pattern of insulin concentration in the culture medium after glucose stimulation interval day was similar throughout the 7 days cultivation. In summary, experimental evidences indicated that the effects of alginate-microencapsulation prolonged survival of the neonatal porcine islets in vitro cultures and the insulin response to glucose of the islets was maintained.

Anti-Proliferative Effects of Dendrophthoe pentandra Methanol Extract on BCR/ABL-Positive and Imatinib-Resistant Leukemia Cell Lines

  • Zamani, Afiqah;Jusoh, Siti Asmaa Mat;Al-Jamal, Hamid Ali Nagi;Sul'ain, Mohd Dasuki;Johan, Muhammad Farid
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.11
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    • pp.4857-4861
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    • 2016
  • Background: Imatinib mesylate, a tyrosine kinase inhibitor specifically targeting the BCR/ABL fusion protein, induces hematological remission in patients with chronic myeloid leukemia (CML). However, the majority of CML patients treated with imatinib develop resistance with prolonged therapy. Dendrophthoe pentandra (L.) Miq. is a Malaysian mistletoe species that has been used as a traditional treatment for several ailments such as smallpox, ulcers, and cancers. Methods: We developed a resistant cell line (designated as K562R) by long-term co-culture of a BCR/ABL positive CML cell line, K562, with imatinib mesylate. We then investigated the anti-proliferative effects of D. pentandra methanol extract on parental K562 and resistant K562R cells. Trypan blue exclusion assays were performed to determine the IC50 concentration; apoptosis and cell cycle analysis were conducted by flow cytometry. Results: D. pentandra extract had greater anti-proliferative effects towards K562R ($IC50=192{\mu}g/mL$) compared to K562 ($500{\mu}g/mL$) cells. Upon treatment with D. pentandra extract at the IC50. concentration: K562 but not K562R demonstrated increase in apoptosis and cell cycle arrest in the G2/M phase. Conclusion: D. pentandra methanol extract exerts potent anti-proliferative effect on BCR/ABL positive K562 cells.

Endothelial Cell Products as a Key Player in Hypoxia-Induced Nerve Cell Injury after Stroke

  • Cho, Chul-Min;Ha, Se-Un;Bae, Hae-Rahn;Huh, Jae-Taeck
    • Journal of Korean Neurosurgical Society
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    • v.40 no.2
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    • pp.103-109
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    • 2006
  • Objective : Activated endothelial cells mediate the cascade of reactions in response to hypoxia for adaptation to the stress. It has been suggested that hypoxia, by itself, without reperfusion, can activate the endothelial cells and initiate complex responses. In this study, we investigated whether hypoxia-induced endothelial products alter the endothelial permeability and have a direct cytotoxic effect on nerve cells. Methods : Hypoxic condition of primary human umbilical vein endothelial cells[HUVEC] was induced by $CoCl_2$ treatment in culture medium. Cell growth was evaluated by 3,4,5-dimethyl thiazole-3,5-diphenyl tetrazolium bromide [MTT] assay Hypoxia-induced products [$IL-1{\beta},\;TGF-{\beta}1,\;IFN-{\gamma},\;TNF-{\alpha}$, IL-10, IL-6, IL-8, MCP-l and VEGF] were assessed by enzyme-linked immunosorbent assay. Endothelial permeability was evaluated by Western blotting. Results : Prolonged hypoxia caused endothelial cells to secrete IL -6, IL -8, MCP-1 and VEGF. However, the levels of IL -1, IL -10, $TNF-{\alpha},\;TGF-{\beta},\;IFN-{\gamma}$ and nitric oxide remained unchanged over 48 h hypoxia. Hypoxic exposure to endothelial cells induced the time-dependent down regulation of the expression of cadherin and catenin protein. The conditioned medium taken from hypoxic HUVECs had the cytotoxic effect selectively on neuroblastoma cells, but not on astroglioma cells. Conclusion : These results suggest the possibility that endothelial cell derived cytokines or other secreted products with the increased endothelial permeability might directly contribute to nerve cell injury followed by hypoxia.

Antibacterial Activity of Edible Plant against Pathogenic Bacteria 1. Antibacterial Activity of Clove against Staphylococcus aureus (식용식물의 식중독세균에 대한 항균작용 1. Staphylococcus aureus에 대한 Clove의 항균작용)

  • 박찬성
    • Food Science and Preservation
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    • v.5 no.1
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    • pp.89-96
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    • 1998
  • The antibacterial activity of clove (Eugenia caryophyllata Thumb) in culture troth against S. aureus was tested at 35, 5, -20, 50, 55 and 60$^{\circ}C$. Tryptic soy broth(TSB) containing 0∼0.5%(w/v) of clove was inoculated with 105∼107 CFU/ml of S. aureus and incubated at each temperature. The growth of S. aureus occured at 0.1% clove after a prolonged lag period while viable cells of S. aureus decreased more than 2 log cycles at 0.3 and 0.4% clove during 12 hours storage at 35$^{\circ}C$. During 32 days of refrigerated storage at 5$^{\circ}C$, survivors of S. aureus were decreased with the progress of time and increasing clove concentration. At the presence of 0.2, 0.3 and 0.4% clove, bacterial cells were dead after 32, 20 and 16 days of refrigerated storage, respectively. During 32 days of frozen storage at -20$^{\circ}C$, survivors of S. aureus were decreased less than 0.5 log cycle at 0% clove. At the presence of 0.1∼0.4% clove, survivors were decreased 2.5∼3.0 log cycles after 1 day and then decreased 0.4∼0.8 log cycles through the frozen storage. There were small changes in populations of S. aureus in TSB between different concentrations of clove during frozen storage. The D-values of S. aureus at clove 0, 0,2, 0.4% were 28.53, 15.14, 8.9 min at 50$^{\circ}C$, 18.43, 10.32, 6.74 min at 55$^{\circ}C$ and 12.78, 9.88, 5.72 min at 60$^{\circ}C$, respectively. The D-values for S. aureus were decreased with the increasing temperature and clove concentration.

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29-kDa FN-f inhibited autophagy through modulating localization of HMGB1 in human articular chondrocytes

  • Hwang, Hyun Sook;Choi, Min Ha;Kim, Hyun Ah
    • BMB Reports
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    • v.51 no.10
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    • pp.508-513
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    • 2018
  • Fibronectin fragments found in the synovial fluid of patients with osteoarthritis (OA) induce the catabolic responses in cartilage. Nuclear high-mobility group protein Box 1 (HMGB1), a damage-associated molecular pattern, is responsible for the regulation of signaling pathways related to cell death and survival in response to various stimuli. In this study, we investigated whether changes induced by 29-kDa amino-terminal fibronectin fragment (29-kDa FN-f) in HMGB1 expression influences the pathogenesis of OA via an HMGB1-modulated autophagy signaling pathway. Human articular chondrocytes were enzymatically isolated from articular cartilage. The level of mRNA was measured by quantitative real-time PCR. The expression of proteins was examined by western blot analysis, immnunofluorescence assay, and enzyme-linked immunosorbent assay. Interaction of proteins was evaluated by immunoprecipitation. The HMGB1 level was significantly lower in human OA cartilage than in normal cartilage. Although 29-kDa FN-f significantly reduced the HMGB1 expression at the mRNA and protein levels 6 h after treatment, the cytoplasmic level of HMGB1 was increased in chondrocytes treated with 29-kDa FN-f, which significantly inhibited the interaction of HMGB1 with Beclin-1, increased the interaction of Bcl-2 with Beclin-1, and decreased the levels of Beclin-1 and phosphorylated Bcl-2. In addition, the level of microtubule-associated protein 1 light chain 3-II, an autophagy marker, was down-regulated in chondrocytes treated with 29-kDa FN-f, whereas the effect was antagonized by mTOR knockdown. Furthermore, prolonged treatment with 29-kDa FN-f significantly increased the release of HMGB1 into the culture medium. These results demonstrated that 29-kDa FN-f inhibits chondrocyte autophagy by modulating the HMGB1 signaling pathway.

Anticancer Effects of Aloe on Sarcoma 180 in ICR Mouse and on Human Cancer Cell Lines (복수암 생쥐와 인체 암세포에 대한 알로에의 항암 작용)

  • Jeong, He-Yun;Kim, Jae-Hyun;Hwang, Se-Jin;Rhee, Dong-Kwon
    • YAKHAK HOEJI
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    • v.38 no.3
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    • pp.311-321
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    • 1994
  • Anticancer effects of Aloe on sarcoma 180 in ICR mouse or human cancer cells were determined. Sarcoma 180 cells were inoculated subcutaneously into male ICR mouse to determine effect of Aloe on tumor gowth, or inoculated intraperitoneally into male ICR mouse to determine effect of Aloe on life span prolongation, followed by oral administration of Aloe vera(10 mg/kg/day, 50 mg/kg/day) or Aloe arborescens(10 mg/kg/day, 100 mg/kg/day) once a day for 14 days. The administration of Aloe vera or Aloe arborescens did not suppress tumor growh. However the life span of ICR mouse was prolonged to 19%(p<0.05), 22%(p<0.05) and 32%(p<0.05) by administration of Aloe vera 10 mg/kg/day, Aloe vera 50 mg/kg/day, and Aloe arborescens 100 mg/kg/day, respectively. To determine anticancer effect of Aloe in vitro, Aloe extract was added to the culture of human gastric cancer cells(SNU-1) and colorectal cancer cells(SNU-C2A), and concentration of Aloe to inhibit cancer cell growth was determined using MTT(3-[ 4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) cytotoxicity assay. High $ID_{50}$ values of Aloe vera and Aloe arborescens against gastric cancer cell line(SNU-1) and colorectal cancer cell line(SNU-C2A) suggest that Aloe gel does not have anticancer effect on these specific human cancer cells although high concentration of Aloe inhibited growth of human cancer cells significantly.

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