• Title/Summary/Keyword: human cells

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Transport Properties of Aromatic Amino Acids by Amino Acid Transporter TAT1 (아미노산 수송체 TAT1에 의한 방향족 아미노산의 수송특성)

  • 김윤배;김명수;윤정훈;박주철;국중기;정해만;최봉규;정규용;김종근
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.775-781
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    • 2002
  • The T-type amino acid transporter 1 (TATI) is a Na$^{+}$-independent amino acid transporter which selectively trans- ports aromatic amino acids subserving the amino acid transport system T. To understand the transport properties of aromatic amino acids by human TAT1 (hTATl ), we have examined the hTATl -mediated aromatic amino acid transports using a Xenopus laeuis oocyte expression system. When expressed in Xenopin laeuis oocytes, hTATl induced L- [$^{14}$ C]tryptophan transport which was not dependent on Na$^{+}$ or Cl$^{[-10]}$ in the medium. Uptake was time-dependent and exhibited a linear dependence on incubation time up to 30 min. The L- ($^{14}$ C)tryptophan uptake was highly inhibited by L-isomers of tryptophan, tyrosine and phenylalanine, whereas other L-amino acids did not inhibit hTATl -mediated L- ($^{14}$ C)tryptophan uptake. The hTATl induced the relatively low-affinity transport of aromatic amino acids such as L- ($^{14}$ C)tryptophan, L- ($^{14}$ C)tyrosine and L- ($^{14}$ C)phenylalanine (Km values: 450~750 $\mu$M), consistent with the properties of classical amino acid transport system T. The L- ($^{14}$ C)tryptophan uptake did not show any remarkable pH dependence within the pH range of 5.5 to 8.5. The time-dependent efflux of L- ($^{14}$ C)tryptophan was detected from the oocytes expressing hTATl, which was not affected by the presence or absence of L-tryptophan in the extracellular medium, indicating that hTATl-mediated transport is due to the facilitated diffusion. Expression of hTATl in Xenopu laevis oocytes induced the transport of tryptophan, tyrosine and phenylalanine, indicating that hTATl is a transporter subserving system T These results suggest that hTATl has essential roles in the absorption of aromatic amino acids from epithelial cells to the blood stream. Hecause hTATl is proposed to be crucial to the efficient absorption of aromatic amino acids from intestine and kidney, its defect such as blue diaper syndrome could be involved in the disruption of aromatic amino acid transport.ort.

Neutrophil Chemotactic Activity in Bronchoalveolar Lavage Fluid of the Rats Exposed to Hyperoxia (고농도의 산소에 노출시킨 쥐의 기관지폐포세척액내 호중구 화학주성활성화도)

  • Song, Jeong Sup;Lee, Sook Young;Moon, Wha Sik;Park, Sung Hak
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.4
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    • pp.547-557
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    • 1996
  • Background : An excessive accumulation of neutrophils in lung tissue has been known to play an important role in mediating the tissue injury among the adult respiratory distress syndrome, idiopathic pulmonary fibrosis and cystic fibrosis by releasing toxic oxygen radicals and proteolytic enzymes. Therefore, it is important to understand a possible mechanism of neutrophil accumulation in lung tissue. In many species, exposure to hyperoxic stimuli can cause changes of lung tissues very similar to human adult respiratory distress syndrome and neutrophils are also functioning as the main effector cells in hyperoxic lung injury. The purpose of the present study was to examine whether neutrophils function as a key effector cell and to study the nature of possible neutrophil chemotactic factors found in bronchoalveolar lavage fluid from the hyperoxia exposed rats. Methods : We exposed the rats to the more than 95% oxygen for 24, 48, 60 arid 72 hours and bronchoalveolar lavage(BAL) was performed. Neutrophil chemotactic activity was measured from the BAT- fluid of each experimental groups. We also evaluated the molecular weight of neutrophil chemotactic tractors using fast performance liquid chromatography and characterized the substances by dialyzer membrane and heat treatment. Results : 1) The neutrophil proportions in bronchoalveolar lavage fluid began to rise from 48 hours after oxygen exposure, and continued to be significantly increased with exposure times. 2) chemotactic index for neutrophils in lung lavages from rats exposed to hyperoxia was significantly higher in 48 hours group than in control group, and was significantly increased with exposure time. 3) No deaths occured until after 48 hours of exposure. However, mortality rates were increased to 33.3 % in 60 hours group and 81.3 % in 72 fours group. 4) Gel filtration using fast performance liquid chromatography disclosed two peaks of neutrophil chemotactic activity in molecular weight of 104,000 and 12,000 daltons. 5) Chemotactic indices of bronchoalveolar lavage fluid were significantly deceased when bronchoalveolar lavage fluid was treated with heat ($56^{\circ}C$ for 30 min or $100^{\circ}C$ for 10 min) or dialyzed (dialyzer membrane molecular weight cut off : 12,000 daltons). Conclusion : These results suggested that the generation of neutrophil chemotactic factor and subsequent neutrophil influx into the lungs are playing an important roles in hyperoxia-induced acute lung injury. Neutrophil chemotactic factor in the lung lavage fluids consisted of several distinct components having different molecular weight and different physical characteristics.

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Fermentation Properties and Increased Health Functionality of Kimchi by Kimchi Lactic Acid Bacteria Starters (김치 유산균 Starter를 이용한 김치의 발효 특성 및 기능성 증진 효과)

  • Bong, Yeon-Ju;Jeong, Ji-Kang;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1717-1726
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    • 2013
  • Fermentation characteristics and health functionalities of kimchi by inoculating kimchi lactic acid bacteria (LAB) starters were studied. We manufactured single LAB starter kimchi (Lactobacillus plantarum pnuK, Lactobacillus plantarum 3099K, Leuconostoc mesenteroides pnuK), mixed LAB starter kimchi (Lb. plantarum pnu/Leu. mesenteroides pnuK, Lb. plantarum 3099/Leu. mesenteroides pnuK) with inoculum size of $10^6$ CFU/g, as well as naturally fermented kimchi (NK), and fermented them for 6 days at $15^{\circ}C$. The pH and acidity of the early phase of fermentation were not different, but kimchi with the starters showed rapid changes in the pH and acidity from 2 days of fermentation. As the fermentation progressed, the level of total aerobic bacteria and Lactobacillus sp. increased similarly with or without Lb. plantarum (LP) inoculation. However, the level of Leuconostoc sp. was high in kimchi inoculated with Leuconostoc sp. starter. In the sensory evaluation test, kimchi with starters received higher overall acceptability scores than those of NK; mixed starter added kimchi earned the highest score. In DPPH and hydroxyl radical scavenging activity, kimchi with the starters exhibited higher activity than that of NK. In the MTT assay of HCT-116 and HT-29 human colon cancer cells, NK showed inhibition rates of 63.4 and 51.9%, but LPpnuK achieved 77.1 and 68.8%, respectively. This study showed that inoculating starters in kimchi increased in vitro antioxidant and anticancer activities, and single starter (LP) added kimchi revealed higher functionality than the kimchi with mixed starter. Kimchis with the starters effectively up-regulated the gene expressions of the pro-apoptotic gene of Bax, but down-regulated Bcl-2. They promoted expressions of p53 and p21, and suppressed expressions of inflammation-related genes, iNOS and COX-2, compared with NK. Taken together, it is expected that using starters may help manufacture kimchi with improved sensory quality and health functionality.

Effects of Fermentation on the Metabolic Activities of Pine Needle Juice (발효과정이 솔잎 착즙액의 항산화, alpha-Glucosidase 및 Angiotensin Converting Enzyme 저해 활성에 미치는 영향)

  • Kim, So-Yun;Lee, Hyun-Jung;Park, Jae-Hee;Kim, Rae-Young;Cheong, Hyeonsook;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.325-334
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    • 2013
  • The objective of this study was to compare the content and metabolic activities between fresh pine needle juice (PNJ) and fermented pine needle juice (FPNJ). A variety of factors were measured, including total phenolic content (TPC), antioxidant activity [DPPH radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), cellular antioxidant capacity (CAC)], anti-genotoxic activity, ${\alpha}$-glucosidase inhibitory activity, and angiotensin converting enzyme (ACE) inhibitory activity. The TPC was $17.3{\pm}0.2$ and $4.6{\pm}0.0$ mg GAE/g in PNJ and FPNJ, respectively. The DPPH RSA, TRAP, and ORAC values increased in a dose-dependent manner for both PNJ and FPNJ, with significantly higher activities in PNJ than FPNJ. The CAC against AAPH-induced oxidative stress in HepG2 cells was protected by both PNJ and FPNJ. Pretreatment with PNJ and FPNJ in human leukocytes produced significant reductions in $H_2O_2$-induced DNA damage at a concentration of $50{\mu}g/mL$. ${\alpha}$-Glucosidase inhibitory activity was significantly higher in FPNJ than PNJ. The ACE inhibitory activity was about 87.1% and 60.0% in 1:1 diluted PNJ and FPNJ, respectively. This study suggests that the fermentation of PNJ could enhance the regulation of blood glucose metabolism and both PNJ and FPNJ might be a new potential source of natural antioxidant, anti-diabetic, and anti-hypertensive agents applicable to food.

Effects of Schizandra chinensis Extract on the Growth of Intestinal Bacteria Related with Obesity (오미자 추출물이 비만과 관련된 장내 세균의 생육에 미치는 영향)

  • Jeong, Eun-Ji;Lee, Woon-Jong;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.673-680
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    • 2009
  • This study was conducted to screen for plant food materials that improve human intestinal microflora, especially microorganisms associated with obesity. Among 30 tested plant food materials, the extract of Schizandra chinensis inactivated Eubacterium limosum, Bacteroides fragilis and Clostridium spp. Additionally, S. chinensis extract was also found to have a growth-promoting effect on Bifidobacterium spp.. The antimicrobial activity and antioxidant activity of the water extract did not decrease in respond to heating. Additionally, the water extract of S. chinensis did not show a toxic effect on the growth of Caco-2 cells. In vivo feeding tests were performed to investigate the influence of extract on the intestinal microflora in rats. Although the extract did not reduce obesity induced by a high fat diet, it led to significant increase in the population of Bifidobacterium spp. and a decrease in the population of Clostridium spp. in rats. Taken together, these results indicate that S. chinensis could be useful as a functional food component to control intestinal microbial flora.

Cutaneous hydration effect of collagen hydrolysate containing collagen tripeptides (콜라겐 트리펩타이드를 고함량으로 함유하는 콜라겐 가수분해물의 피부 보습 효과)

  • Kim, Ae-Hyang;Kim, Yi-Soo;Piao, Zhe;Shin, Yong Chul;Ha, Min Woo
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.420-429
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    • 2018
  • Skin ageing is associated with compromised performance of its fundamental barrier functions, with undesirable changes in appearance. Since this may introduce a detrimental impact on the quality of life, significant effort to discover effective ingredients against ageing is being invested. Recently, collagen hydrolysates containing tripeptides such as GlyPro-Hyp (GPH) have been developed with anticipation of improved effects compared to that of existing collagen hydrolysate-products. To evaluate the cutaneous hydration effect of collagen tripeptides (CTP), meaningful biomarkers in human dermal fibroblasts (HDF) and NC/Nga Tnd mice were analyzed in this study. Increased levels of ceramide kinase, hyaluronic acid, collagen 1A, and hyaluronan synthase-2 (HAS2), and decreased levels of hyaluronidase-1 (HYAL1) and CD44 in HDF cells were demonstrated. Furthermore, significant reduction of transepidermal water loss (TEWL), scratching behavior, HYAL1, $TNF-{\alpha}$ and IL-6 and increased water content and HAS2 were verified by in vivo tests. These results strongly suggest the potential of CTP as a skin hydration agent.

A Study of Peripheral Blood and Bone Marrow Responses Depends on the Frequency of rhG-CSF Administration in Dogs (개에서 재조합 과립구 자극 인자 (rh G-CSF)의 투여 간격에 따른 말초혈액과 골수의 반응에 대한 연구)

  • Kim, Ji-Hyun;Jee, Cho-Hee;Won, Jin-Hee;Jung, Hae-Won;Moon, Jong-Hyun;Cho, Kyu-Woan;Kang, Byeong-Teck;Jung, Dong-In
    • Journal of Veterinary Clinics
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    • v.31 no.2
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    • pp.77-84
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    • 2014
  • The present study evaluated that responses of peripheral and bone marrow depends on the frequency of recombinant human granulocyte colony-stimulating factor (rhG-CSF) administration in dogs. The rhG-CSF has been revealed that have a beneficial effect for dogs with myelosuppression secondary to chemotherapy or radiation but there were no studies about the frequency of administration in dogs. In this research, rhG-CSF was administrated $5{\mu}g/kg$ subcutaneously for each two-dogs group as follows: (1) every day for trial, (2) every other day for trial, (3) every third day for trial. The peripheral blood analysis including direct microscopic differential counts of one hundred cells was performed every day. Bone marrow aspiration was performed before administration of rh G-CSF, on the day of 0, 3, 9 and when the WBC counts were decreased within the normal range (day 12 or 13). Rh G-CSF was well-tolerated and showed no side effects in all dogs. According to the present study, $5{\mu}g/kg$ administration of rhG-CSF have cell-specific, frequency-related effect on bone marrow and peripheral blood. Furthermore, the effects of rhG-CSF administration on bone marrow sustained during the study and prolonged at least 3 days after discontinuing of rhG-CSF treatment.

Antitumor and Immuno-potentiating Activities of Crude Polysaccharides from Fruiting Body of Agaricus brasiliensis (신령버섯(Agaricus brasiliensis) 자실체 추출 조다당류의 항암 및 면역증강 작용)

  • Cha, Youn-Jeong;Kim, Jeong-Hwa;Lee, Tae-Soo;Lee, U-Youn
    • The Korean Journal of Mycology
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    • v.39 no.1
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    • pp.57-67
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    • 2011
  • Agaricus brasiliensis, one of edible mushroom belonging to Basidiomycota, has been used for curing gastric ulcer and stomach cancer of human beings and also known to have good inhibitory effects on sarcoma 180 and Ehrlich carcinoma of mice. Neutral saline soluble (0.9% NaCl), hot water soluble and methanol soluble substances (hereinafter referred to Fr. NaCl, Fr. HW and Fr. MeOH, respectively) were prepared from fruiting body of the mushroom. ${\beta}$-glucan and total protein contents were identify from fractions of edible mushrooms extract. The ${\beta}$-glucan and protein contents of all fractions of the mushrooms ranged from 21.54~32.31% and 0.16~9.34%, respectively. In vitro cytotoxicity tests, crude polysaccharides were not cytotoxic against cancer cell lines such as Sarcoma 180, HT-29, NIH3T3 and RAW 264.7 at the concentration of 10~2000 ${\mu}g/ml$. Intraperitoneal injection with crude polysaccharides exhibited life prolongation effect of 18.8~50.6% in mice previously inoculated with Sarcoma 180. Fr. HW increased the numbers of spleen cell by 1.2 fold at the concentration of 200 ${\mu}g/ml$ compared with control. Fr. MeOH and Na improved the immuno-potentiating activity of B lymphocyte by increasing the alkaline phosphatase activity by 1.6 fold compared with control at the concentration of 50~500 ${\mu}g/ml$. Fr. Na generated 15.9 ${\mu}M$ of nitric oxide (NO) when cultured with RAW 264.7 at the concentration of 200 ${\mu}g/ml$, while lipopolysaccharide, a positive control, produced 3.7 ${\mu}M$. The Fr. NaCl, Fr. HW and Fr. MeOH increased the secretion of TNF-${\alpha}$, IL-$1{\beta}$, Il-2 and IL-6 by 2.2 times compared with the control group. Fr. Na increased the numbers of peritoneal exudate cells by 4 folds at the concentration of 50mg/kg compared with control. Circulating leukocytes increased by 2.7 folds when Fr. HW from A. brasiliensis was inoculated at the concentration of 50 mg/kg body weight. The hematological and blood chemical analysis of the 3 fractions did not show any difference in blood compositions and enzyme activities compared with the control group (p<0.05). Therefore, the experimental results suggested that crude polysaccharides extracted from A. brasiliensis contain antitumor and immuno-potentiating activities against Sarcoma 180 in ICR mice.

Effects of Ibandronate on the Expression of Matrix Metalloproteinases in Human U2OS Osteosarcoma Cells (사람 U2OS 골육종 세포에서 Matrix Metalloproteinase의 발현에 Ibandronate가 미치는 영향)

  • Jung, Sung-Taek;Seo, Hyoung-Yeon;Xin, Zeng-Feng;Kim, Yang-Kyung;Kim, Hyung-Won
    • The Journal of the Korean bone and joint tumor society
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    • v.15 no.2
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    • pp.111-121
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    • 2009
  • Background: Osteosarcoma is one of the most common primary malignant tumors of bone occurring mainly in children and adolescents. Although surgery combined with chemotherapy has markedly improved patient survival during the last years, the use of anticancer drugs is still associated with serious problem, such as the frequent acquisition of drug-resistant phenotypes and occurrence of "secondary malignancies". Several solid tumors display enhanced expression of matrix metalloproteinases (MMPs), and recently clinical trials have been initiated on MMP-inhibitors. On the other hand, bisphosphonates (BPs) are inhibitors of bone resorption, and widely used to treat osteoclast-mediated bone diseases. Also they appear to possess direct antitumor activity. Methods: One osteosarcoma cell line (U2OS) was treated with ibandronate (0, 0.1, 1, $10{\mu}M$) for 48 hours. Cell viabilities were determined using MTT assay, the mRNA levels of MMP-2 and MT1-MMP were detected by reverse-transcription polymerase chain reaction, the amount of MMP-2 and MT1-MMP protein were measured by Westernblot, the activities of MMP-2 were observed by Gelatin zymography, and Matrigel invasion assays were used to investigate the invasive potential of osteosarcoma cell lines before and after ibandronate treatment. Results: The invasiveness of U2OS cell line was reduced dose-dependently following 48 hour treatment of up to $10{\mu}M$ of the ibandronate at which concentration no cytotoxicity occurred. Furthermore, the gelatinolytic activities and protein and mRNA levels of MMP-2 and MT1-MMP were also suppressed by increasing ibandronate concentrations. Conclusion: Given that MMP-2 is instrumental in tumor cell invasion, it is very likely that the reduction in osteosarcoma cell invasion by ibandronate is a consequence, at least in part, of suppressed expression of both MMP-2 and MT1-MMP. Isolation of a molecule (s) responsible for the bisphosphonate inhibition of tumor cell invasion would pave the way for the development of a new generation of metastasis inhibitors.

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Evaluation of the Cell-Mediated Immunity in Treatment Failure Pulmonary Tuberculosis (치료실패 폐결핵 환자의 세포성면역반응에 관한 연구)

  • Park, Jeong-Kyu;Park, Jang-Seo;Kim, Hwa-Jung;Jo, Eun-Gyeong;Min, Dul-Lel;Lim, Jae-Hyun;Suhr, Ji-Won;Paik, Tae-Hyun
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.1
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    • pp.13-25
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    • 1999
  • Background: Ineffective cell-mediated immune response in human tuberculosis is associated with a depressed Thl cytokine response and reduced production of IFN-$\gamma$. Most persons infected with Mycobacterium tuberculosis are healthy tuberculin reactors with protective immunity, but a minority with ineffective immunity develop extensive pulmonary tuberculosis. The cell-mediated immune response is an important aspect of host resistance to mycobacterial infection and is believed to be tightly regulated by a balance between Th1 cytokines including IFN-$\gamma$, IL-12, IL-18, regulated on activation, normal T cell expressed and secreted (RANTES) and Th2 counterparts such as IL-4, monocyte chemoattractant protein-l (MCP-l). Methods: Proliferation and mRNA expression of IFN-$\gamma$, RANTES and MCP-l by RT-PCR in peripheral blood mononuclear cells (PBMCs) in response to in vitro stimulation with mycobacterial antigens were compared in pulmonary tuberculosis patients with cured and treatment failure and in tuberculin-positive and tuberculin-negative healthy subjects. Results: Defective proliferative responsiveness to aqueous TSP antigen was involved with treatment failure tuberculosis patients. Aqueous TSP antigen-induced IFN-$\gamma$ and RANTES mRNA expression was decreased in treatment failure tuberculosis patients compared with healthy tuberculin reactors and cured tuberculosis patients (23.1 % versus 90.0% for IFN-$\gamma$ and 46.2% versus 70.0% versus 46.2% for RANTES). The frequency of MCP-l mRNA expression to aqueous TSP antigen in treatment failure tuberculosis patients was greater than in healthy tuberculin reactors and cured tuberculosis patients (76.9% versus 40.0%). Conclusion: The increasing expression of MCP-1 mRNA in response to aqueous TSP antigen might be predicted to favor Th1 responses and restricted Th1 responses in treatment failure of pulmonary tuberculosis.

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