• Title/Summary/Keyword: Serum deprivation

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Effects of Serum on Nitric Oxide Production in Embryonic Mouse Liver Cell Line BNL CL.2 (혈청이 마우스 간 세포주 BNL CL.2의 Nitric Oxide 생성에 미치는 영향)

  • 김유현;김신무;배현옥;유지창;정헌택;진효상
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.85-93
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    • 1999
  • Nitric oxide (NO) plays an important role in immunologic defense, and influences upon the functioning of secretory tissues and cells. It also exhibits cytotoxic/cytostatic activity as one of major operating effectors of the cellular immunity system. We investigated the effects of serum on the cell damages and NO production in the mouse liver cell line BNL CL.2 to establish the role of NO. We observed that, when BNL CL.2 cells were cultured in serum-free medium, they were induced to cell damage by the stimulation of IFN-$\gamma$ alone or IFN-$\gamma$ plus LPS. Serum-starved cells showed large amount of nitrite accumulation and NO synthase (NOS) expression in response to IFN-$\gamma$ alone in dose- and time- dependent manners, but serum-supplied cells did not The production of NO was blocked by protein tyrosine kinase (PTK) inhibitors, genistein and herbimycin. These results suggest that the deprivation of serum in the BNL CL.2 cell culture medium might primed with the cells to produce NO when the cells are triggered by IFN-$\gamma$ and the involvement of PTK signal transduction pathway in the expression of NOS gene in murine hepatocytes.

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Modulation of Cell Proliferation and Hypertrophy by Gangliosides in Cultured Human Glomerular Mesangial Cells

  • Lee Seoul;Ahn Seon Ho;Baek Seung Hoon;Song Ju Hung;Choo Young Kug;Kwon Oh Deog;Choi Bong Kyu;Jung Kyu Yong
    • Archives of Pharmacal Research
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    • v.28 no.8
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    • pp.948-955
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    • 2005
  • Glomerular mesangial cells (GMCs) in diverse renal diseases undergo cell proliferation and/or hypertrophy, and gangliosides have been reported to play an important role in modulating cell structure and function. This study compared the effects of transforming growth $factor-\beta\; (TGF­\beta1)$ and the effects of the application of exogenous gangliosides on GMCs and investigated whether the application of exogenous gangliosides regulated cellular proliferation and hypertrophy. Human GMCs were cultured with exogenous gangliosides and $TGF-\beta1$ in a media containing $10\%$ fetal bovine serum and in a media without the fetal bovine serum. Exogenous gangliosides biphasically changed the proliferation of human GMCs (0.1-1.0 mg/mL). A low concentration (0.1 mg/mL) of gangliosides mainly increased the number of human GMCs, whereas cellular proliferation was significantly reduced by raising the concentration of exogenous gangliosides. $TGF-\beta1$ greatly reduced the number of human GMCs in a concentration­dependent manner (1-10 ng/mL). Serum deprivation accelerated the gangliosides- and $TGF­\beta1-induced$ inhibition of mesangial cell proliferation to a greater extent. Gangliosides (1.0 mg/ mL) and $TGF-\beta1$ (10 ng/mL) both caused a significant increase in the incorporation of $[^3H]leucine$ per cell in the serum-deprived condition, whereas it was completely reversed in serum­supplemented condition. Similar results to the $[^3H]leucine$ incorporation were also observed in the changes in cell size measured by flow cytometric analysis. These results show that exogenous gangliosides modulate cell proliferation and hypertrophy in cultured human GMCs, and these cellular responses were regulated differently based on whether the media contained serum or not. Results from the present study raise new possibilities about the potential involvement of gangliosides in the development of mesangial cell proliferation and hypertrophy.

Expression and Purification of Recombinant Human Angiopoictin-2 and Its Analog in Chinese Hamster Ovary Cells

  • Hwang, Su-Jeong;Kim, In-Jun;Go, Gyu-Yeong;Lee, Gyun-Min
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.459-462
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    • 2001
  • Angiopoietin-2 (Ang2) is a naturally occurring antagonist for angiopoietin-l (Angl) and its Tie2 receptor during vasculogenesis, Although angiopoietins have been expressed in several mammalian cell lines, their expression levels are low. Recombinant Chinese hamster ovary (CHO) cell lines expressing a high level of human Ang2 or its analog, human $Ang2_{443}$, with an amino-terminal FLAG-tag were constructed by transfecting the expression vectors into dhfr-deficient CHO cells and subsequent gene amplification in medium containing stepwise increments in methotrexate level. Secreted Ang2 or human $Ang2_{443}$ were purified from the cultured medium using an anti-FLAG- agarose affinity chromatography, The purified Ang2 and $Ang2_{443}$ migrated on SOS-PAGE as a broad band, characteristic of glycosylated protein. Their biological activity in vitro was demonstrated in a serum deprivation-induced apoptosis assay. Ang2 at high concentration, like AngI, can be an apoptosis survival factor for endothelial cells through the activation of the Tie2 receptor.

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Recent Advances in the Relationship between Endocrine Status and Nutrition in Chickens - Review -

  • Okumura, J.;Kita, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.7
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    • pp.1135-1141
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    • 1999
  • A large number of investigations have shown that changes in nutritional condition affect endocrine status in avian species. Herein, recent findings including novel peptides discovered by the development of the techniques in the field of molecular biology have been reviewed. The insulin-like growth factors (IGF-I and IGF-II) found in chickens have been characterized and shown to be 70 and 66 amino acid polypeptides, respectively. Plasma IGF-I level is very responsive to nutrition, Le. varying dietary proteins and energy intakes, and food restriction. Plasma IGF-II concentration is altered by nutritional deprivation to a much smaller extent than plasma IGF-I concentration. Almost all of the serum and tissue IGFs are found in a complex composed of IGF and IGF-binding protein (IGFBP). In the chicken plasma, the major IGFBP differs from that in mammalian plasma. The proglucagon mRNA encodes glucagon and two glucagon-like peptides (GLP-I and GLP-2). The intracerebroventricular administration of GLP-l strongly decreased food intake of chicks, and it was indicated that the inhibition of food intake by GLP-l was associated with neuropeptide Y, which is one of the neurotransmitters reported to enhance food intake.

Effects of Soy Hydrolysate Fractions on Appetite Suppression and Ghrelin Releasing in ICR Mice (ICR 마우스를 대상으로 대두 가수분해물 분획물의 식욕 억제 및 Ghrelin 분비에 대한 연구)

  • Jung, Eun Young;Suh, Hyung Joo
    • The Korean Journal of Food And Nutrition
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    • v.27 no.2
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    • pp.225-230
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    • 2014
  • The objective of this study was to investigate the effects of soy hydrolysate fractions on appetite suppression and ghrelin releasing. In a short-term experiment, the cumulative food intake and serum ghrelin level were decreased significantly (p<0.05) during a 4-hr period after the interperitoneal injection of soy hydrolysate fractions (0.5, 1 g/kg BW), following a 12-hr period of food deprivation. In a long-term experiment, food efficiency ratio (FER) was also reduced significantly (p<0.05), when soy hydrolysate fractions (0.5, 1% in drinking water) were given orally for 8 wks. Therefore, we found that soy hydrolysate fractions affected food intake through appetite and ghrelin releasing in short-term and long-term experiments. In conclusion, this study indicated that soy hydrolysate fractions would diminish the sensation of hunger by reducing the secretion of orexigenic factors such as ghrelin that send satiety signals to the brain, terminating food intake.

The Effect of Autophagy to Cell Death in Nutrient-Deprived H460 Cells (영양분이 결핍된 H460 세포주에서 자가포식이 세포사멸에 미치는 영향)

  • Jang, Hye-Yeon;Jo, Hyang-Jeong;Hwhang, Ki-Eun;Kim, So-Young;Lee, Kang-Kyoo;Moon, Sun-Rock;Shin, Jeong-Hyun;Cho, Kyung-Hwa;Lee, Mi-Kung;Lee, Sam-Youn;Park, Soon-Ah;Park, Jong-Kun;Kim, Hui-Jung;Yang, Sei-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.69 no.2
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    • pp.81-94
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    • 2010
  • Background: Autophagy is an important adaptive mechanism in normal development and in response to changing environmental stimuli in cancer. Previous papers have reported that different types of cancer underwent autophagy to obtain amino acids as energy source of dying cells in nutrient-deprived conditions. However, whether or not autophagy in the process of lung cancer causes death or survival is controversial. Therefore in this study, we investigated whether nutrient deprivation induces autophagy in human H460 lung cancer cells. Methods: H460, lung cancer cells were incubated in RPMI 1640 medium, and the starved media, which are BME and RPMI media without serum, including 2-deoxyl-D-glucose according to time dependence. To evaluate the viability and find out the mechanism of cell death under nutrient-deprived conditions, the MTT assay and flow cytometry were done and analyzed the apoptotic and autophagic related proteins. It is also measured the development of acidic vascular organelles by acridine orange. Results: The nutrient-deprived cancer cell is relatively sensitive to cell death rather than normal nutrition. Massive cytoplasmic vacuolization was seen under nutrient-deprived conditions. Autophagic vacuoles were visible at approximately 12 h and as time ran out, vacuoles became larger and denser with the increasing number of vacuoles. In addition, the proportion of acridine orange stain-positive cells increased according to time dependence. Localization of GFP-LC3 in cytoplasm and expression of LC-3II and Beclin 1 were increased according to time dependence on nutrient-deprived cells. Conclusion: Nutrient deprivation induces cell death through autophagy in H460 lung cancer cells.

Final height of Korean patients with early treated congenital hypothyroidism

  • Lee, Jiyun;Lee, Jeongho;Lee, Dong Hwan
    • Clinical and Experimental Pediatrics
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    • v.61 no.7
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    • pp.221-225
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    • 2018
  • Purpose: Congenital hypothyroidism (CH) is the most common endocrine disorder in children. Thyroid hormone deprivation results not only in mental retardation but also growth retardation. This study investigates the final height (FH) in Korean patients with CH detected by newborn screening and examines factors that may affect the FH. Methods: The medical records of Korean CH patients (n=45) were reviewed. The FH was examined and target height (TH) was calculated based on mid-parental height. The FH z score (FHZ) and TH z score (THZ) were computed using the 2007 Korean National Growth Chart. The FHZ and THZ were compared with a Student t test. The impact of the etiology of CH (athyreosis, dyshormonogenesis, ectopic thyoid, hypoplastic thyroid), initial serum thyroid stimulating hormone (TSH) level, initial free thyroxine (T4) level, and time of therapy initiation based on FH was assessed. Results: The mean FHZ was $0.10{\pm}1.01$ for male patients and $-0.11{\pm}1.09$ for female patients. There were no significant differences between FHZ and THZ for both female (P=0.356) and male patients (P=0.237). No significant relationship was found between FH and the etiology of CH, initial TSH level, initial free T4 level, and the time of therapy initiation. Conclusion: Early intervention and satisfactory management do not appear to impede growth in Korean patients with CH. Thus, early detection and proper management of patients with CH detected by newborn screening program are necessary.

Molecular Taxonomy of a Soil Actinomycete Isolate, KCCM10454 Showing Neuroprotective Activity by 16S rRNA and rpoB Gene Analysis

  • Lee Bong Hee;Kim Hong;Kim Hyun Ju;Lim Yoon Kyu;Byun Kyung Hee;Hutchinson Brian;Kim Chang Jin;Ko Young Hwan;Lee Keun Hwa;Cha Chang Yong;Kook Yoon Hoh;Kim Bum Joon
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.213-218
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    • 2005
  • Epilepsy constitutes a significant public health problem, and even the newest drugs and neurosurgical techniques have proven unable to cure the disease. In order to select a group of isolates which could generate an active compound with neuroprotective or antiepileptic properties, we isolated 517 actinomycete strains from soil samples taken from Jeju Island, in South Korea. We then screened these strains for possible anti-apoptotic effects against serum deprivation-induced hippocampal cell death, using the 3-(4, 5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide (MTT) assay as an in vitro test. The excitotoxic glutamate analog, kainic acid (KA), was used to induce seizures in experimental mice in our in vivo tests. As a result of this testing, we located one strain which exhibited profound neuroprotective activity. This strain was identified as a Streptomyces species, and exhibited the rifampinresistant genotype, Asn$(AAC)^$442, according to the results of 16S rRNA and rpoB gene analyses

Identification of Three Competitive Inhibitors for Membrane­Associated, $Mg^{2+}-Dependent$ and Neutral 60 kDa Sphingomyelinase Activity

  • Kim Seok Kyun;Jung Sang Mi;Ahn Kyong Hoon;Jeon Hyung Jun;Lee Dong Hun;Jung Kwang Mook;Jung Sung Yun;Kim Dae Kyong
    • Archives of Pharmacal Research
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    • v.28 no.8
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    • pp.923-929
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    • 2005
  • Methanol extracts of domestic plants of Korea were evaluated as a potential inhibitor of neutral pH optimum and membrane-associated 60 kDa sphingomyelinase (N-SMase) activity. In this study, we partially purified N-SMase from bovine brain membranes using ammonium sulfate. It was purified approximately 163-fold by the sequential use of DE52, Butyl-Toyopearl, DEAE-Cellulose, and Phenyl-5PW column chromatographies. The purified N-SMase activity was assayed in the presence of the plant extracts of three hundreds species. Based on the in vitro assay, three plant extracts significantly inhibited the N-SMase activity in a time- and concentration-dependent manner. To further examine the inhibitory pattern, a Dixon plot was constructed for each of the plant extracts. The extracts of Abies nephrolepis, Acer tegmentosum, and Ginkgo biloba revealed a competitive inhibition with the inhibition constant (Ki) of $11.9 {\mu}g/mL,\;9.4{\mu}g/mL,\;and\;12.9{\mu}g/mL$, respectively. These extracts also inhibited in a dose-dependent manner the production of ceramide induced by serum deprivation in human neuroblastoma cell line SH-SY5Y.

Bicalutamide-induced Interstitial Lung Disease (전립선 암에서 Bicalutamide 사용으로 생긴 간질성 폐질환 1예)

  • Kim, Yang-Kyun;Kim, Yee-Hyung;Lee, Jae-Jin;Choi, Cheon-Woong;Yoo, Jee-Hong;Park, Myung-Jae;Kang, Hong-Mo
    • Tuberculosis and Respiratory Diseases
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    • v.68 no.4
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    • pp.226-230
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    • 2010
  • Androgen deprivation therapy, which is the standard treatment for metastatic prostate cancer, includes nonsteroidal antiandrogenic drugs, such as flutamide, nilutamide and bicalutamide. Of them, bicalutamide rarely induces interstitial pneumonia. We report a case of bicalutamide-induced interstitial pneumonia. A 68-year old male diagnosed with prostate cancer and multiple bone metastases presented with dry cough and low grade fever for 3 days. He had taken bicalutamide (50 mg/day) for 13 months. High resolution computed tomography revealed ground glass opacity in his right upper lung. The laboratory studies showed no eosinophilia in the serum and bronchoalveolar lavage fluid. Despite the use of antimicrobial agents for 2 weeks, the extent of the lung lesions increased to the left upper and right lower lung. He had no environmental exposure, collagen vascular disease and microbiological causes. Under the suspicion of bicalutamide-induced interstitial pneumonia, bicalutamide was stopped and prednisolone (1 mg/kg/ day) was initiated. The symptoms and radiologic abnormalities were resolved with residual minimal fibrosis.