• 제목/요약/키워드: culture-dependent

검색결과 1,198건 처리시간 0.029초

Expression of SDF-$1{\alpha}$ and leptin, and their effect on expression of angiogenic factors in mouse ovaries

  • Park, Min-Jung;Park, Sea-Hee;Lee, Su-Kyung;Moon, Sung-Eun;Moon, Hwa-Sook;Joo, Bo-Sun
    • Clinical and Experimental Reproductive Medicine
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    • 제38권3호
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    • pp.135-141
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    • 2011
  • Objective: Ovarian angiogenesis plays an important role in folliculogenesis. However, little is known about the expression of angiogenic factors during follicular development according to female age. Stromal cell derived factor-$1{\alpha}$ (SDF-$1{\alpha}$) plays a role in granulosa cell survival and embryo quality as an angiogenic chemokine. Leptin is also involved in folliculogenesis and angiogenesis. This study examined expression of SDF-$1{\alpha}$ and leptin, and their effects on the expression of angiogenic factors in the ovary during follicular development according to female age. Methods: Ovaries were collected from C57BL mice of two age groups (6-9 weeks and 24-26 weeks) at 6, 12, 24, and 48 hours after 5 IU pregnant mare's serum gonadotropin (PMSG) injection. The expression of ovarian SDF-$1{\alpha}$ and leptin mRNA was evaluated by RT-PCR. In the organ culture experiment, the ovaries were cultured in transwell permeable supports with Waymouth's medium treated with various doses of SDF-$1{\alpha}$(50-200 ng/mL) or leptin (0.01-1 ${\mu}g$/mL) for 7 days. Then, mRNA expression of vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), and visfatin were examined in the cultured ovaries. Results: Expression of SDF-$1{\alpha}$ and leptin in the ovary was significantly lower in the aged mouse group compared to the young mouse group ($p$ <0.05). Expression of these two factors increased with follicular development after PMSG administration. SDF-$1{\alpha}$ treatment stimulated visfatin expression in a dose-dependent manner, while leptin treatment significantly increased eNOS expression. Conclusion: These results suggest that decrease of ovarian SDF-$1{\alpha}$ and leptin expression may be associated with aging-related reduction of ovarian function. SDF-$1{\alpha}$ and leptin may play a role in follicular development by regulating the expression of angiogenic factors in mouse ovaries.

주택 담보 가계 대출액 결정요인 추정에 관한 패널 데이터 모형 연구 (Estimating the Determinants of Loan Amount of Housing Mortgage : A Panel Data Model Approach)

  • 김희철;신현철
    • 한국컴퓨터정보학회논문지
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    • 제16권7호
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    • pp.183-190
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    • 2011
  • 주택담보 가계 대출은 그룹(지역)별, 시간별로 다양한 원인에 의해서 가계대출 결정요인이 이루어지고 있어 복잡성을 띠고 있다. 본 연구에서는 복잡성을 띠고 있는 주택담보 가계 대출에 관련된 제 변인들을 파악하기 위해 패널 데이터를 이용한 연구 모형을 설정하고 이를 통해 가계대출에 결정적으로 영향을 미치는 제 변인에 대하여 조사, 분석, 검증한다. 본 연구는 7 그룹(6개 광역시(부산, 대구, 인천, 광주, 대전, 울산) 및 서울)을 분석대상으로 하였다. 분석기간은 2007년 1월부터 2010년 9월 까지 자료를 이용하였고. 주택담보 가계 대출액을 종속변수로 설정하고 소비자물가지수, 실업률, 가구당 월평균가계소득, 보건의료비 지출률, 종합주가지수, 일반은행 가계 대출연체율을 설명(독립)변수로 투입하였다. 주택담보 가계 대출 요인을 추정한 결과 소비자물가지수와 실업률은 정(+)의 영향을 미치는 유의한 변인으로 나타나고 보건 의료비 지출률은 음(-)의 영향을 나타내는 유의적인 변인으로 나타났다. 그러나 가구당월평균 가계소득액, 종합주가지수와 일반은행 가계대출 연체율은 비유의적인 변인으로 나타나 주택담보 가계 대출에는 큰 영향을 주지는 않은 것으로 나타났다.

DMB 영상콘텐츠 제작기법의 최적화에 대한 연구 (Study on the Optimization of DMB Image Contents Production Method)

  • 임평종;김종서;곽훈성
    • 디지털콘텐츠학회 논문지
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    • 제9권3호
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    • pp.399-412
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    • 2008
  • 21세기는 정보 지향과 정보 의존이 심화되는 지식 기반 사회로 변화하고 있다. 뉴미디어 시대의 방송은 방송과 통신의 융합이 보편화되어 새로운 형태의 방송 서비스가 가능해짐에 따라 수용자의 요구를 충족시키는 소비자 중심의 서비스 형태로 변화되고 있다. 단방향 송신만 하던 기존 방송과 달리 양방향 커뮤니케이션이 가능한 퍼스널 미디어 시대가 등장한 것이다. 그 결과 새로운 복합매체들이 상용화 되었지만 대표적인 경우가 DMB이다. DMB는 나만의 TV 혹은 내손안의 TV라고 말한다. 하지만 콘텐츠 측면으로는 기존 방송의 재전송이라는 한계를 드러내고 있다. 향후의 DMB 방송은 재전송에서 벗어나 그 특성에 맞는 콘텐츠로 제작되어야만 할 것이다. DMB 콘텐츠의 제작방향을 정립하기 위해서는 먼저 DMB라는 매체의 특성 및 서비스의 영역을 정확하게 분석하는 것이 필요하다. 본 연구는 이러한 분석을 통해 DMB에 적합한 프로그램 개발과 적정한 시간, 카메라 앵글과 워킹 등의 시각적 표현방법 등 전반적인 영상콘텐츠의 최적화된 제작기법에 대해 제안하고자 한다. DMB의 특성에 맞게 제작된 콘텐츠는 수용자들로 하여금 더욱 친숙하게 느끼게 할 것이며 나아가 새로운 시청 문화로까지 자리 잡을 것으로 기대한다.

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반복 유산환자의 말초혈액 단핵구와 태반항원을 체외 공동 배양시 세포 매개 면역반응에 프로게스테론이 미치는 영향 (The Effects of Progesterone on Cell Mediated Immunity to Trophoblast in Woman with Recurrent Pregnancy Loss)

  • 최범채
    • Clinical and Experimental Reproductive Medicine
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    • 제24권3호
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    • pp.399-405
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    • 1997
  • Progesterone is necessary for successful pregnancy and had immunosuppressive properties. Peripheral blood mononuclear cells (PBMC) from many women with unexplained recurrent spontaneous abortion responded to trophoblast extract in vitro by prolifertion and releasing soluble, heat-labile factors that are toxic to mouse embryos (embryotoxic factors). Accumulating evidence suggests that T Helper (Th)-1 type immunity to trophoblast is correlated with embryotoxic factor production and is associated with pregnancy loss, while Th2-type immunity is associated with successful gestation. The objective of this study was to determine whether progesterone can inhibit Th1-type cytokine secretion (IFN-${\gamma}$, TNF-${\alpha}$) by trophoblast-activated peripheral blood mononuclear cells from 23 nonpregnant women (age 25-35) with unexplained recurrent abortion (median 5, range 3 to 15)who otherwise produce embryotoxic factors in response to trophoblast. We also determined whether progesterone affected Th2-type cytokines (IL-4, IL-10) in this system in vitro and if IL-10 (1,500 pg/mL) could inhibit Th1-type immunity to trophoblast. IFN-${\gamma}$ was detected in 17 of 23 (74%) trophoblast stimulated PBMC culture supernatants ($77.94{\pm}23.79$ pg/mL) containing embryotoxic activity. TNF-${\alpha}$ was detected in 19 (83%) of these same supernatants ($703.15{\pm}131.36$ pg/mL). In contrast, none of the supernatants contained detectable levels of IL-4 or IL-10. Progesterone ($10^{-5}$, $10^{-7}$, $10^{-9}$M) inhibited Th1-type immunity in a dose dependent manner, but had no effect on Th2-type cytokine secretion. The inhibitory effects of progesterone were abrogated with RU486, but did not affect Th2-type cytokine secretion in trophoblast-activated cell cultures. IL-10, like progesterone also inhibited Th1-type cytokine secretion but had no effect on Th2-type cytokines. These data suggest that therapies designed to suppress Th1-type cytokine secretion in women with recurrent abortion who have evidence of Th1-type immunity to trophoblast may be efficacious in preventing pregnancy loss and should be tested in appropriately designed clinical trials.

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Streptococcus thermophilus로 발효한 한약재 발효분말의 항염증 및 미백 효과 (Anti-Inflammatory and Whitening Effect of the Lyophilized Powder of Oriental Plant Extracts Fermented with Streptococcus thermophilus)

  • 최화정;이정희;윤미영;이재숙
    • 대한화장품학회지
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    • 제41권2호
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    • pp.159-164
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    • 2015
  • 새로운 항염증 및 미백 소재를 찾기 위해서, 본 연구에서는 6가지 한약재 추출물(유자, 상백피, 오미자, 율무, 당귀, 고삼)을 Streptococcus thermophilus에 의해 발효한 발효분말의 미백 및 항염증 효과를 피부섬유아세포에 대한 피부독성, 산화질소 생성, tyrosinase 활성, 멜라닌 형성의 저해 효과를 측정함으로써 평가하였다. 발효물은 $37^{\circ}C$에서 2일 동안 Streptococcus thermophilus에 의해 발효한 후, 동결건조에 의해 발효분말을 제조하였다. 발효분말은 피부섬유아세포에 대해 $500{\mu}g/mL$의 농도에서 cytopathic effect reduction 방법을 사용하면서 측정하였을 때 세포독성을 나타내지 않았다. 또한 발효분말은 Griess reagent system을 사용하면서 산화질소 생성에 대한 저해 효과를 나타내었다. 더욱이 발효분말은 tyrosinase 활성에 대한 저해 효과가 농도 의존적으로 나타났다. 발효분말은 배양액으로부터 멜라닌 생성에 대해 유의적인 저해를 나타내었다(p < 0.05). 그러므로 이러한 자료로부터 발효분말은 항염증 및 미백 효과를 갖는 것으로 나타났으며, 화장품을 위한 효과적인 성분으로써 사용가능할 것으로 사료된다.

Endophytic fungi harbored in Panax notoginseng: diversity and potential as biological control agents against host plant pathogens of root-rot disease

  • Zheng, You-Kun;Miao, Cui-Ping;Chen, Hua-Hong;Huang, Fang-Fang;Xia, Yu-Mei;Chen, You-Wei;Zhao, Li-Xing
    • Journal of Ginseng Research
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    • 제41권3호
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    • pp.353-360
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    • 2017
  • Background: Endophytic fungi play an important role in balancing the ecosystem and boosting host growth. In the present study, we investigated the endophytic fungal diversity of healthy Panax notoginseng and evaluated its potential antimicrobial activity against five major phytopathogens causing root-rot of P. notoginseng. Methods: A culture-dependent technique, combining morphological and molecular methods, was used to analyze endophytic fungal diversity. A double-layer agar technique was used to challenge the phytopathogens of P. notoginseng. Results: A total of 89 fungi were obtained from the roots, stems, leaves, and seeds of P. notoginseng, and 41 isolates representing different morphotypes were selected for taxonomic characterization. The fungal isolates belonged to Ascomycota (96.6%) and Zygomycota (3.4%). All isolates were classified to 23 genera and an unknown taxon belonging to Sordariomycetes. The number of isolates obtained from different tissues ranged from 12 to 42 for leaves and roots, respectively. The selected endophytic fungal isolates were challenged by the root-rot pathogens Alternaria panax, Fusarium oxysporum, Fusarium solani, Phoma herbarum, and Mycocentrospora acerina. Twenty-six of the 41 isolates (63.4%) exhibited activity against at least one of the pathogens tested. Conclusion: Our results suggested that P. notoginseng harbors diversified endophytic fungi that would provide a basis for the identification of new bioactive compounds, and for effective biocontrol of notoginseng root rot.

유용식용 균주에 의한 발효 누에분말의 이화학적 특성과 생리활성 (Biological Activity and Biochemical Properties of Silkworm (Bombyx mori L.) Powder Fermented with Bacillus subtilis and Aspergillus kawachii)

  • 차재영;김용순;안희영;강민정;허수진;조영수
    • 생명과학회지
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    • 제21권1호
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    • pp.81-88
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    • 2011
  • 누에 분말을 이용하여 B. subtilis 및 A. kawachii 균주에 의한 고체 발효를 통해 얻어진 발효 누에분말의 발효시간에 따른 이화학적 특성(단백질 함량 및 단백질 분해 패턴) 및 생리활성 효과(항산화 작용, 환원력 및 혈전용해 활성)를 평가하였다. 누에 단백질 함량과 항산화 활성은 B. subtilis 및 A. kawachii 두 균주 발효 모두 12일째에 가장 높았으며, 발효시간 증가와 함께 증가하는 경향을 보였다. 발효 누에분말의 환원력은 B. subtilis 균주 발효에 의해서는 6일째에 A. kawachii 균주 발효에 의해서는 12일째 가장 높은 것으로 나타나 균주간에 차이가 있었다. 혈전 분해 활성은 B. subtilis 및 A. kawachii 두 균주 발효 모두 6일째에 가장 높았으며, 이러한 효과는 B. subtilis 발효 누에분말보다는 A. kawachii 발효 누에분말에서 더 높았다. SDS-PAGE상의 단백질 패턴 분석에서는 66-97 kDa 크기의 단백질 밴드가 B. subtilis 발효에 의해서는 발효 초기인 3일째 대부분 분해가 일어났으나, A. kawachii 발효 누에분말에서는 발효 3일째부터 차츰 분해되기 시작하여 12일째 분해가 대부분 이루어졌다. 따라서 미생물을 이용한 발효 누에에서 특정의 생리활성 작용을 기대하기 위해서는 사용하는 균종에 따라 발효 기간이 달라져야 한다는 것을 시사하였다.

출아효모에서 xylitol dehydrogenase (XYL2)의 최적 생산을 위한 발현 시스템 구축 (Expression System for Optimal Production of Xylitol Dehydrogenase (XYL2) in Saccharomyces cerevisiae)

  • 정회명;김연희
    • 생명과학회지
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    • 제27권12호
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    • pp.1403-1409
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    • 2017
  • 본 연구에서는 lignocellulosic biomass (xylose)의 부가가치를 높이고 효율적인 활용을 위해 xylitol dehydrogenase를 Saccharomyces cerevisiae 숙주세포에서 분비 생산하고자 하였다. 먼저 S. cerevisiae와 Pichia stipitis유래 XYL2 유전자(S.XYL2 and P.XYL2 gene)의 발현 시스템을 구축하기 위하여 GAL10 promoter와 ADH1 promoter 하류에 각각 mating factor ${\alpha}$ ($MF{\alpha}$) signal sequence와 XYL2유전자를 가진 $pGMF{\alpha}-S.XYL2$, $pGMF{\alpha}-P.XYL2$, $pAMF{\alpha}-S.XYL2$$pAMF{\alpha}-P.XYL2$ plasmid를 구축하였다. 각각의 plasmid는 S. cerevisiae $SEY2102{\Delta}trp1$ 균주에 형질전환되었고, 생산된 xylitol dehydrogenase의 활성을 조사해 본 결과, GAL10 promoter가 ADH1 promoter보다 XYL2유전자의 발현에 더욱 적합함을 확인 할 수 있었다. 또한 P. stipitis 유래의 xylitol dehydrogenase 효소 활성이 S. cerevisiae 유래의 효소 활성보다 2배 이상 더 높았으며, 활성의 증가를 위해 두 유전자 모두 cofactor로 $NAD^+$에 의존한다는 것을 확인하였다. 재조합 유전자가 가지는 분비서열에 의해 $SEY2102{\Delta}trp1/pGMF{\alpha}-P.XYL2$ 균주에서 xylitol dehydrogenase의 약 77%는 periplasmic space로 분비 발현되었음을 알 수 있었다. 또한 재조합 xylitol dehydrogenase의 효율적인 생산을 위해 탄소원의 영향을 조사해본 결과, glucose 단독보다 glucose와 xylose를 혼합 배양한 경우에서 효소활성이 최대 41% 정도 증가되었음을 확인 할 수 있었다. 본 연구에서 최적화한 발현 시스템 및 배양 조건은 xylose 뿐만 아니라 다양한 biomass를 이용한 유용물질 생산을 위한 관련 단백질의 발현 분비시스템 구축 및 대량생산에도 응용될 수 있을 것이라 생각된다.

Gut Microbial Metabolites Induce Changes in Circadian Oscillation of Clock Gene Expression in the Mouse Embryonic Fibroblasts

  • Ku, Kyojin;Park, Inah;Kim, Doyeon;Kim, Jeongah;Jang, Sangwon;Choi, Mijung;Choe, Han Kyoung;Kim, Kyungjin
    • Molecules and Cells
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    • 제43권3호
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    • pp.276-285
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    • 2020
  • Circadian rhythm is an endogenous oscillation of about 24-h period in many physiological processes and behaviors. This daily oscillation is maintained by the molecular clock machinery with transcriptional-translational feedback loops mediated by clock genes including Period2 (Per2) and Bmal1. Recently, it was revealed that gut microbiome exerts a significant impact on the circadian physiology and behavior of its host; however, the mechanism through which it regulates the molecular clock has remained elusive. 3-(4-hydroxyphenyl)propionic acid (4-OH-PPA) and 3-phenylpropionic acid (PPA) are major metabolites exclusively produced by Clostridium sporogenes and may function as unique chemical messengers communicating with its host. In the present study, we examined if two C. sporogenes-derived metabolites can modulate the oscillation of mammalian molecular clock. Interestingly, 4-OH-PPA and PPA increased the amplitude of both PER2 and Bmal1 oscillation in a dose-dependent manner following their administration immediately after the nadir or the peak of their rhythm. The phase of PER2 oscillation responded differently depending on the mode of administration of the metabolites. In addition, using an organotypic slice culture ex vivo, treatment with 4-OH-PPA increased the amplitude and lengthened the period of PER2 oscillation in the suprachiasmatic nucleus and other tissues. In summary, two C. sporogenes-derived metabolites are involved in the regulation of circadian oscillation of Per2 and Bmal1 clock genes in the host's peripheral and central clock machineries.

Effects of Insulin and IGFs on Phosphate Uptake in Primary Cultured Rabbit Renal Proximal Tubule Cells

  • Han, Ho-Jae;Park, Kwon-Moo
    • The Korean Journal of Physiology
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    • 제30권1호
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    • pp.63-76
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    • 1996
  • The aim of present study was to characterize phosphate uptake and to investigate the mechanism for the insulin and insulin-like growth factor(IGF) stimulation of phosphate uptake in primary cultured rabbit renal proximal tubule cells. Results were as follows : 1. The primary cultured proximal tubule cells had accumulated $6.68{\pm}0.70$ nmole phosphate/mg protein in the presence of 140 mM NaCl and $2.07{\pm}0.17$ nmole phosphate/mg protein in the presence of 140 mM KCl during a 60 minute uptake period. Raising the concentration of extracellular phosphate to 100 mM$(48.33{\pm}1.76\;pmole/mg\;protein/min)$ induced decrease in phosphate uptake compared with that in control cells maintained in 1 mM phosphate$(190.66{\pm}13.01\;pmole/mg\;protein/min)$. Optimal phosphate uptake was observed at pH 6.5 in the presence of 140 mM NaCl. Phosphate uptake at pH 7.2 and pH 7.9 decreased to $83.06{\pm}5.75%\;and\;74.61{\pm}3.29%$ of that of pH 6.5, respectively. 2. Phosphate uptake was inhibited by iodoacetic acid(IAA) or valinomycin treatment $(62.41{\pm}4.40%\;and\;12.80{\pm}1.64%\;of\;that\;of\;control,\;respectively)$. When IAA and valinomycin were added together, phosphate uptake was inhibited to $8.04{\pm}0.61%$ of that of control. Phosphate uptake by the primary proximal tubule cells was significantly reduced by ouabain treatment$(80.27{\pm}6.96%\;of\;that\;of\;control)$. Inhibition of protein and/or RNA synthesis by either cycloheximide or actinomycin D markedly attenuated phosphate uptake. 3. Extracellular CAMP and phorbol 12-myristate 13 acetate(PMA) decreased phosphate uptake in a dose-dependent manner in all experimental conditions. Treatment of cells with pertussis toxin or cholera toxin inhibited phosphate uptake. cAMP concentration between $10^{-6}\;M\;and\;10^{-4}\;M$ significantly inhibited phosphate uptake. Phosphate uptake was blocked to about 25% of that of control at 100 ng/ml PMA. 3-Isobutyl-1-methyl-xanthine(IBMX) inhibited phosphate uptake. However, in the presence of IBMX, the inhibitory effect of exogenous cAMP was not significantly potentiated. Forskolin decreased phosphate transport. Acetylsalicylic acid did not inhibit phosphate uptake. The 1,2-dioctanoyl-sn-glycorol(DAG) and 1-oleoyl-2-acetyl-sn- glycerol(OAG) showed a inhibitory effect. However, staurosporine had no effect on phosphate uptake. When PMA and staurosporine were treated together, inhibition of phosphate uptake was not observed. In conclusion, phosphate uptake is stimulated by high sodium and low phosphate and pH 6.5 in the culture medium. Membrane potential and intracellular energy levels are also an important factor fer phosphate transport. Insulin and IGF-I stimulate phosphate uptake through a mechanisms that involve do novo protein and/or RNA synthesis and decrease of intracellular cAMP level. Also protein kinase C(PKC) is may play a regulatory role in transducing the insulin and IGF-I signal for phosphate transport in primary cultured proximal tubule cells.

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