• Title/Summary/Keyword: ER beta

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Overexpression of Cyclin E and its Low Molecular Weight Isoforms Cooperate with Loss of p53 in Promoting Oncogenic Properties of MCF-7 Breast Cancer Cells

  • Montazeri, Hamed;Bouzari, Saeid;Azadmanesh, Kayhan;Ostad, Seyed Nasser;Ghahremani, Mohammad Hossein
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7575-7582
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    • 2015
  • Cyclin E, a key coordinator of the G1 to S transition in the cell cycle, may be deregulated in several malignancies, including breast cancer. The most significant aberration in cyclin E is its elastase mediated proteolytic cleavage into tumor specific low molecular weight isoforms (LMW-Es). LMW-Es are biochemically hyperactive and biologically drive tumorigenesis in transgenic mouse models. Additionally, expression of LMW-Es has been correlated with poor survival in breast cancer cases. Here we determine whether expression of LMW-Es in a breast cancer cell line that is naturally devoid of these deregulated forms would alter their progression through each phase of the cell cycle. The results revealed that LMW-Es expression resulted in an increased doubling time, concomitant with a predominant increase in the population in the S phase of the cell cycle. Moreover, downregulation of p53 in LMW-Es cells resulted in additional shortening of the doubling time and enrichment of cells in the S and G2/M phases of the cell cycle. Furthermore, expression of LMW-Es sensitized cells to ${\beta}$-estradiol (E2) mediated growth and changed expression patterns of estrogen receptor and Bcl-2. Intriguingly, expression of LMW-Es could surpass anti-apoptotic effects raised by p53 upregulation. Taken together these studies suggest that overexpression of LMW-Es in collaboration with p53 loss results in altered g rowth properties of MCF-7 cells, enhancing the oncogenic activity of these ER positive breast cancer cells.

Transcriptional Activity of an Estrogen Receptor β Subtype in the Medaka Oryzias dancena

  • Maeng, Sejung;Yoon, Sung Woo;Kim, Eun Jeong;Nam, Yoon Kwon;Sohn, Young Chang
    • Development and Reproduction
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    • v.23 no.4
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    • pp.333-344
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    • 2019
  • In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, fold-activation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.

Angelica polymorpha Maxim Induces Apoptosis of Human SH-SY5Y Neuroblastoma Cells by Regulating an Intrinsic Caspase Pathway

  • Rahman, Md. Ataur;Bishayee, Kausik;Huh, Sung-Oh
    • Molecules and Cells
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    • v.39 no.2
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    • pp.119-128
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    • 2016
  • Angelica polymorpha Maxim root extract (APRE) is a popular herbal medicine used for treating stomachache, abdominal pain, stomach ulcers, and rheumatism; however the effect of APRE on cancer cells has not yet been explored. Here, we examined APRE cytotoxicity seen on target neuroblastoma cells (NB) using cell viability assays, DAPI visualization of fragmented DNA, and Western blotting analysis of candidate signaling pathways involved in proliferation and apoptosis. We demonstrated that APRE reduced cell viability in NB to a greater extent than in fibroblast cells. In addition, we found that APRE could inhibit the three classes of MAPK proteins and could also down-regulate the PI3K/AKT/GSK-$3{\beta}$ activity all being relevant for proliferation and survival. APRE could also up-regulate Bax expression and down-regulate Bcl-2 and Mcl-1. With APRE treatment, depolarization of mitochondria membrane potential and activation of caspase-3 was demonstrated in the SH-SY5Y cells. We could not found increased activity of death receptor and caspase-8 as markers of the extrinsic apoptosis pathway for the APRE treated cells. In presence of a caspase-3 siRNA and a pan-caspase inhibitor, APRE could not reduce the viability of NB cells to a significant degree. So we predicted that with APRE, the intrinsic pathway was solely responsible for inducing apoptosis as we also showed that the non-caspase autophagy pathway or ER stress-ROS mediated pathways were not involved. These findings demonstrate that an intrinsic mitochondria-mediated apoptosis pathway mediates the apoptotic effects of APRE on SH-SY5Y cells, and that APRE shows promise as a novel agent for neuroblastoma therapy.

Anwendungsbereich der Verleitung des Patienten im Sinne des ${\S}27$ Abs. 3 das Gesuntheitsdienstgesetz (의료법 제27조 제3항 환자 '유인' 금지의 적용범위)

  • Lee, Seok-Bae
    • The Korean Society of Law and Medicine
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    • v.12 no.1
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    • pp.11-39
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    • 2011
  • [ ${\S}27$ ]Abs. 3 das Gesuntheitsdienstgesetz (the Medical Service Act) in Korea lautet: Niemand in der Absicht, sich oder einem Dritten einen $Verm{\ddot{o}}gensvorteil$ zu verschaffen, der Medizininstitut bzw. dem Mediziner (die Medizinerin) den Patienten vorstellen, ${\ddot{u}}bweweisen$, verleiten oder einen anderen zu dieser Handlung anstiften darf, wie z.B. die Selbstbeteiligung des Patienten nach dem Krankenkassengesetz (the National Health Insurance Act) oder dem Gesetz ${\ddot{u}}ber$ Beistand der ${\ddot{a}}rztlicher$ Betreuung (the Medical Care Assistance Act) skontieren oder befreien, Geld offerieren oder dem Allgemeinheit das Verkehrswesen anbieten usw. Nach dem Wortlaut ist jedoch unklar, ob unter diese Vorschriften der Fall subsumiert werden kann, wenn eine Medizininstitut bzw. ein(e) Mediziner(in) in der Absicht, sich einen $Verm{\ddot{o}}gensvorteil$ zu verschaffen, sich den Patienten verleitet. Nach dem Korean Supreme Court ist eine Medizininstitut bzw. ein(e) Mediziner(in) nur dann das Subjekt der Verleitungshandlung, wenn sie bzw. er ein Mittel gegen fairen oder $ordungsm{\ddot{a}}{\beta}ien$ Medizinmarkt verwendet oder dem Patienten eine ${\ddot{a}}rztlich$ rechtswidrige Behandlung (z.B. einen rechtswidrigen Schwangerschaftsabbruch) verspricht. In diesem Beitrag wird dagegen die Auffassung mittels der teleologischen Reduktion vertritt und argumentiert, dass ein ${\ddot{a}}rztlich$ rechtswidriges Behandlung nach dem Rechtsgut und dem Normzweck unter ${\S}27$ Abs. 3 das Gesuntheitsdienstgesetz nicht subsumiert werden, sondern allein nach eigenem Unrecht bestraft werden kann.

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A Comparative Study of Protein Profiles in Porcine Fetus Fibroblast Cells with Different Confluence States

  • Han, Rong-Xun;Kim, Hong-Rye;Diao, Yunfei;Kim, Myung-Youn;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.243-248
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    • 2009
  • To examine the differential expression of proteins during the cycling (70~80% confluences) and G0/G1 (full confluences) phases in porcine fetal fibroblast cells, we used a global proteomics approach by 2-D gel electrophoresis (2-DE) and MALDI-TOF-MS. Cycling cell were harvested at approximately 70% to 80% confluent state while cells in G0/G1 phase were recovered after maintenance of a confluent state for 48 hr. Cellular proteins with isoelectric points ranging between 3.0~10.0, were analyzed by 2-DE with 2 replicates of each sample. A total of approximately 700 spots were detected by 2.D gels stained with Coomassie brilliant blue. On comparing the cell samples obtained from the cycling and G0/G1 phases, a total of 13 spots were identified as differentially expressed proteins, of which 8 spots were up-regulated in the cycling cell and 5 were up-regulated in the G0/G1 phase. Differentially expressed proteins included K3 keratin, similar to serine protease 23 precursor, protein disulfide-isomerase A3, microsomal protease ER-60, alpha-actinin-2, and heat-shock protein 90 beta. The identified proteins were grouped on the basis of their basic functions such as molecular binding, catabolic, cell growth, and transcription regulatory proteins. Our results show expression profiles of key proteins in porcine fetal fibroblast cells during different cell cycle status.

Induction of In Vitro Vitellogenin Synthesis by Bisphenol, Nonylphenol and Octylphenol in Chinese Minnow(Phoxinus oxycephalus) Hepatocytes

  • Park, Chang-Beom;Kim, Byung-Ho;Na, Oh-Soo;Choi, Young-Chan;Lee, Young-Don;Baek, Hae-Ja;Kim, Hyung-Bae;Akihiro Takemura
    • Animal cells and systems
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    • v.7 no.3
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    • pp.227-235
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    • 2003
  • Bisphenol A (BPA), nonylphenol (NP), and 4-tert-octylphenol (OP) are known endocrine disrupting chemicals (EDCs) with estrogenic activity in fish. This study compared the effects of BPA, NP and OP on in vitro vitellogenin (VTG) synthesis in primary cultures of hepatocytes of the Chinese minnow Phoxinus oxycephalus. The VTG secreted into the culture medium was measured using enzyme-linked immunosorbent assay (ELISA), which we developed in this study using an antibody prepared from homogenates of Chinese minnow egg. VTG synthesis was induced by estradiol-17$\beta$ ($E_2$) and phenols (BPA, NP and OP) treatment. $E_2$ at concentrations of 10$^{-6}$ M or higher increased VTG levels significantly (P < 0.05). Exposure to 10^5\;M\;BPA\;or\;10^-4$M NP and OPinduced in vitro VTG synthesis (P < 0.01). However, $10^-3$ M BPA, NP or OP did not induce VTG synthesis. These results suggest that SPA has the highest estrogenic potential in Chinese minnow hepatocytes. Tamoxifen, an anti-estrogen, drastically blocked the production of VTG by phenols (BPA, NP and OP) suggesting that phenols (BPA, NP and OP) may act via binding to estrogen receptor (ER) in Chinese minnow hepatocytes.

Sex Steroid Hormone and Ophthalmic Disease (성호르몬과 안질환)

  • Kim, Jin-Ju;Yu, Hyeong-Gon;Ku, Seung-Yup
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.2
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    • pp.89-98
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    • 2010
  • Sex and its tropic hormones influence the lacrimal system, corneal anatomy and disease, aqueous humor dynamics and glaucoma, crystalline lens and cataract, and retinal disease. Dry eye occurs especially frequently during pregnancy, oral contraceptive use, and after menopause, during which androgen levels decrease. Androgen control development, differentiation, and lipid production of sebaceous glands throughout the body, and androgen deficiency also leads to meibomian gland dysfunction and evaporative dry eye. On the other hand, estrogen causes a reduction in size, activity, and lipid production of sebaceous glands. Sex and its tropic hormones also influence the corneal anatomy and disease, and corneal thickening occurred on the second day of the menstrual cycle and around the time of ovulation and appeared to be related to estrogen levels. Fuchs' dystrophy is more commonly seen in postmenopausal women than men and may be linked to hormonal changes that occur with aging. In addition, overexpression of estrogen and progesterone receptors in the conjunctiva of vernal keratoconjunctivitis patients. Serum progesterone levels also may be associated with intraocular pressure especially in pregnant women, and for the women. For women with cataracts, hormone levels were typical of menopause, and there was a significant negative correlation between estradiol and follicular stimulating hormone levels. In addition, serum testosterone levels are associated with the development of diabetic retinopathy. Although the role of sex hormones on the eye is largely unknown, and the results should be interpreted with caution until replicated, the functions of sex hormones in ocular disease remains to be investigated, because they may be involved in structure and function of the ocular components, which are important in the pathogenesis of ocular disease.

Genenation of structural diversity in polyketides by combinatorial biosynthesis of polyketides: Part I. Generation of multiple bioactive macrolides by hybrid modular polyketide synthases in Streptomyces venezuelae, Part II. Production of novel rifamycins by combinatorial biosynthesis

  • Yoon, Yeo-Joon
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2002.10a
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    • pp.18-25
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    • 2002
  • The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

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Molecular Modification of Perilla Lipid Composition

  • Hwang, Young-Soo;Kim, Kyung-Hwan;Hwang, Seon-Kap;Lee, Sun-Hwa;Lee, Seong-Kon;Kim, Jung-Bong;Park, Sang-Bong;Tom Okita;Kim, Donghern
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.20-30
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    • 1999
  • In order to modify lipid production of Perilla qualitatively as well as quantitatively by genetic engineering, genes involved in carbon metabolism were isolated and characterized. These include acyl-ACP thioesterases from Perilla frutescens and Iris sp., four different $\beta$-ketoacyl- ACP synthases from Perilla frutescens, and two $\Delta$15 a-cyl-ACP desaturases(Pffad7, pffad3). Δ15 acyl-ACP desa turase (Δ15-DES) is responsible for the conversion of linoleic acid (18:2) to $\alpha$-linolenic acid (ALA, 18:3). pffad 3 encodes Δ15 acyl-desaturase which is localized in ER membrane. On the other hand, Pffad7 encodes a 50 kD plastid protein (438 residues), which showed highest sequence similarity to Sesamum indicum fad7 protein. Northern blot analysis revealed that the Pffad7 is highly expressed in leaves but not in roots and seeds. And Pffad3 is expressed throughout the seed developmental stage except very early and fully mature stage. We constructed Pffad7 gene under 355 promoter and Pffad3 gene under seed specific vicillin promoter. Using Pffad7 construct, Perilla, an oil seed crop in Korea, was transformed by Agrobacterium leaf disc method. $\alpha$-linolenic acid contents increased in leaves but decreased in seeds of transgenic Perilla. Currently, we are transforming Perilla with Pffad3 construct to change Perilla seed oil composition. We isolated three ADP-glucose pyrophosphorylase (AGP) genes from Perilla immature seed specific cDNA library. Nucleotide sequence analysis showed that two of three AGP (Psagpl, Psagp2) genes encode AGP small subunit polypeptides and the remaining (Plagp) encodes an AGP large subunit. PSAGPs, AGP small subunit peptide, form active heterotetramers with potato AGP large subunit in E. coli expressing plant AGP genes.

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데데킨트 절단, 배중률, 관계

  • Hong, Seong-Gi
    • Korean Journal of Logic
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    • v.7 no.2
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    • pp.15-46
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    • 2004
  • Um die rationalen Zahlen auf die reellen Zahlen zu erweitern und dadurch die Stetigkeit der reellen Zahlen sicherzustellen, hat der deutsche Mathematiker R. Dedekind im Jahr 1872 in seinem Aufsatz "Stetigkeit und Irrationale Zahlen" einen neuen mathmatischen Begriff $eingef\ddot{u}hrt,\;n\ddot{a}mlich$ 'Schnitt'. Die Menge aller rationalen Zahlen Q wird durch eine rationale Zahl a zu zwei Untermengen $A_1=\{x|x{\leq}a,\;x{\in}Q\}$, $A_2=\{x|x>a,\;x{\in}Q\}$ $vollst\ddot{a}ndig$ geteilt. Wenn wir solche Teilung, d.i. solchen Schnitt mit "$(A_1,\;A_2)$" bezeichnen, ist ein $Identit\ddot{a}tssatz$ "a=$(A_1,\;A_2)$" absolut harmlos. Analog dazu glaubt Dedekind fest, $da{\beta}$ jede irrationale Zahl mit Hilfe von einem entsprechenden Schnitt $einzuf\ddot{u}hren$ ist. Zum Beispiel, falls die zwei Mengen $B_1=\{x|x^2<2,\;x{\in}Q\}$ und $B_2=\{x|x^2>2,\;x{\in}Q\}$ gegeben sind, dann $w\ddot{a}re$ die irrationale Zahl $^{\surd}2$ mit $(B_1,\;B_2)$ gleichzusetzen. Im Fall von einem Schnitt der Menge der rationalen Zahlen durch eine rationale Zahl, $(A_1,\;A_2)$, haben die beiden Untermengen $A_1$ und $A_2$ jwewils ein Supremum und ein Infimum und beide $m\ddot{u}ssen$ identisch sein, aber -wie schon Russell in seinem Buch "Introduction to Mathmatical Philosophy" dies kritisiert- hat ein Schnitt $f\ddot{u}r$ die $Einf\ddot{u}hrung$ der irrationalen Zahl, $(B_1,\;B_2)$ keine solche $gl\ddot{u}cklichen$ Eigenschaften. Dennoch glaubt Dedekind an eine streng wissenschaftliche Fundierung der irrationalen Zahl fest, und $w\ddot{u}rde$ nach dem Grund seines Glaubens befragt, $k\ddot{o}nnte$ er nur seine Behauptung wiederholen, ein klarer Fall circulus vitiosus. Mit anderen Worten, die $L\ddot{u}cke$ zwischen $B_1$ und $B_2$ durch die $Einf\ddot{u}hrung$ einer [einzigen] wissenschaftlich fundierten irrationalen Zahl $\ddot{u}berbr\ddot{u}ckt$ und das Ganze zu einem Kontinium gemacht werden sollte, bleibt dieses Vorhaben von Dedekind erst als eine Hoffnung und dessen Resultat kann $h\ddot{o}chstens$ nur als ein Postulat, aber keineswegs als ein methodisch einwandfreier Beweis betrachtet werden. Die Probleme, die mit dem Versuch der $Einf\ddot{u}hrung$ der irrationalen Zahlen mit Hilfe von Schnitt verbunden sind, sind nicht spezifisch allein im Gebient der Mathmatik, sondern betreffen immer wieder die Rechtfertigungsfrage der $Einf\ddot{u}hrung$ der letzten Bestandteile im bezug auf eine Systemerstellung, egal ob dies System ein Wissenschaftliches oder unsere $allt\ddot{a}gliche$ Sprachhandlung ist. $F\ddot{u}r$ all diese Rechtfertigungsfragen gilt das in der klassischen Logik $g\ddot{a}ngige$ logische Prinzip tertium non datur nicht mehr, aber nicht nur wegen der von praktischen $Unm\ddot{o}glichkeit$, die unendlichen vielen $Gegenst\ddot{a}nde$ durchforschen zu $m\ddot{u}ssen$, das $hei{\beta}t$, wegen der erkenntnistheoretischen $Beschr\ddot{a}nktheit$ des jetzigen Erkenntnisniveau, sondern auch wegen des speziellen ontologischen modus der $eizuf\ddot{u}hrenden$ Objekten. Der Autor des Aufsatzes analysiert $\ddot{a}nliche$ $F\ddot{a}lle$ (das Urmeterbeispiel und die Chrakterisierungen der geometrischen Axiome von Wittgenstein), und versucht mit Hilfe von beiden Begriffen, 'interne' und 'externe' Relation, zu zeigen, $da\beta$ eine gemeinsame, invariante Struktur in den eben genannten $F\ddot{a}llen$ besteht. Am Ende des Aufsatzes setzt der Autor sich mit der logischen Argumentationsstruktur des Zitates tiber 'Grenze' aus Noten von Leonardo da Vinci auseinander, und weist auf einen $m\ddot{o}glichen$ Zusammenhang der Grundidee seienes Aufsatzes mit der Philosophie der indischen Denker $N\bar{a}g\bar{a}rjuna$ hin, obwohl die zitierten Versen aus dem Hauptwerk von $N\bar{a}g\bar{a}rjuna$, dem Mittleren Weg$(Madhyamakak\bar{a}rik\bar{a})$ nur andeutend sein $m\ddot{o}gen$.

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