• BMB Reports
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• 제28권5호
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• pp.392-397
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• 1995

The effect of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on the intracellular $Ca^{2+}$ level was studied in NIH3T3 fibroblast cells. A reduction of the intracellular $Ca^{2+}$ level was observed after exposure to 300 ${\mu}m$ MNNG. However, the intracellular level of $IP_3$, a well-known regulator of $Ca^{2+}$ release from internal storage, was not changed by MNNG treatment. Instead, a reduction of the intracellular NAD level was observed. NAD as well as $IP_3$ stimulated intracellular $Ca^{2+}$ release from permeabilized cells. The treatment of 3-aminobenzamide, which inhibited the MNNG-induced reduction of the NAD level, also prevented the MNNG-induced decrease of the $Ca^{2+}$ level. Our data suggest that MNNG reduces the intracellular $Ca^{2+}$ level by NAD depletion in NIH3T3 cells.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.136-140
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• 2000

A sensitive and quantiative method for the structural analysis of oligosaccharide was established for the glycoform analysis of glconproproteins. Inthis study, n-linked oligosaccharides of human IgG and bovine transferin were analyzed for the evaluation of the methydrate moiety ofthe method. Chrbohydrate moiety of glycoprotein was relased by hydrazinolysis and purified by paper chromatography. The oligosaccharides were labeled with a fluorescent bye, 2-aminobenzamide, for the enhancement of detection sensitivity. sialylated (acidic) oligosaccharides were separated from neutral oligosaccharide by employing a strong anion-exchange column(MonoQ) followed by the treatment with sialidase. Enzymatically desiayated fractions and neutral fractions of oligosaccharides were applied to normal-phase HPLC to resolve the peaks according to glucose unit (GU). The structure of separated molecules was further determined by sequential digestion with exoglycosidases. As a result, disialylated biantennary complextype oligosaccharide was found to be a major sugar chain in bovine transferrin (63%). In human IgG, core fucosylated asialobiantennary complex oligosaccharides were dominant. These results coincided well with reported results.

• 약학회지
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• 제19권2호
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• pp.96-100
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• 1975

2-Ethoxy-4-acetamidobenzoate (III) was synthesized by ethylation of N-formyl compound obtained with formylation of methyl 4-aminosalicylate and 2-ethoxy-4-acetamidobenzamide (VIII) was synthesized by ethylation of 4-acetamidosalicylamide (VII) formed by reacting methyl 4-acetamido-2-acetoxy benzoate with concentrated ammonia water under pressure. 2-Ethoxy-4-acetamido benzamide (VIII) was also synthesized by acetylation of 2-ethoxy-4-aminobenzamide (VI) obtained by pressure eraction of methyl 2-ethoxy-4-formylamido-benzoate (I) with concentrated ammonia water.

• 대한한의학회지
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• 제22권3호
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• pp.21-30
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• 2001

목적 : 이전 연구에서 단삼 추출액이 강력한 항산화 작용이 있음을 확인한 바 있어 단삼 추출액이 신장세뇨관 세포에서 oxidant에 의한 세포사망 및 DNA 손상을 방지하는 지를 조사하고 이러한 효과가 지질의 과산화를 억제하는 효과에 기인하는 지를 시험하였다. 방법 : 신장 근위세뇨관 세포 유래 세포주인 opossum kidney (OK)세포를 이용하여 세포 사망은 frypan blue exclusion방법을 이용하여 평가하였고, DNA손상 정도는 double stranded DNA의 파괴를 측정하여 평가하였다. Oxidant 약물 모델로는 $H_2O_2$를 사용하였다. 결과 : $H_2O_2$는 적용시 간과 농도에 비례하여 세포 사망을 유도하였다. 단삼 추출액은 0.05% 농도에서 $H_2O_2$에 의한 세포사망 및 DNA 손상을 방지하였다. 이러한 방지효과는 $H_2O_2$ 제거 효소인 catalase와 철 착염제인 deferoxamine에 의해서도 나타났다. 그러나 강력한 항산화제인 DPPD는 $H_2O_2$에 의한 세포 사망이나 DNA손상을 방지하지 못하였다. $H_2O_2$는 세포내 ATP 농도를 감소시켰으며. 이러한 감소는 poly (ADP-ribose) polymerase억제제인 3-aminobenzamide에 의해 방지되었으나 단삼 추출액에 의해서는 영향을 받지 않았다. 3-aminobenzamide는 $H_2O_2$에 의한 세포 사망을 방지하였다. $H_2O_2$는 지질의 과산화를 증가시켰으며, 이러한 변화는 단삼 추출액과 DPPD에 의해 방지되었다 결론 : OK 세포에서 $H_2O_2$에 의한 세포사망과 DNA 손상에는 지질의 과산화가 중요한 역할을 하지 않으며, 단삼 추출액의 $H_2O_2$에 의한 세포 사망과 DNA 손상 방지 효과는 항산화 작용이 아닌 다른 기전에 기인하는 것으로 사료된다.

• 대한임상검사과학회지
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• 제43권4호
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• pp.161-170
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• 2011

Quercetin is one of the most distributed flavonoids in the plant kingdom and occurs naturally in a wide range of fruits and vegetables. This study was undertaken to determine whether quercetin exerts beneficial effect against necrotic and apoptotic cell death induced by hydrogen peroxide ($H_2O2$) in intestinal cells using the human-derived cultured T84 colonic epithelial cell line. Necrotic cell death was induced by exposing cells to 0.5 mM $H_2O_2$ for 2 h and apoptosis was induced by incubating cells in normal culture medium for 18 h following exposure of cells to 0.5 mM $H_2O2$ for 2 h. Cell viability was evaluated by the trypan blue exclusion assay and apoptosis was assessed by Hoechst 33258 staining and flow cytometry. $H_2O_2$ induced necrotic cell death in a time and dose-dependent fashion. Both necrotic and apoptotic cell deaths were not prevented by the antioxidants N,N'-diphenyl-p-phenylenediamine(DPPD) and Trolox, whereas both cell deaths induced by the organic hydroperoxide t-butylhydroperoxide (tBHP) were prevented by DPPD, suggesting that $H_2O_2$ induces cell death through a lipid peroxidation-independent mechanism. $H_2O2$-induced necrotic death was prevented by deferoxamine and 3-aminobenzamide, while the apoptotic cell death was not affected by these agents. Quercetin prevented both necrotic and apoptotic cell deaths induced by $H_2O_2$ in a dose-dependent manner. $H_2O_2$ caused activation of poly (ADP-ribose) polmerase (PARP), which was inhibited by deferoxamine, 3-aminobenzamide, and quercetin, but not DPPD. These results indicate that quercetin inhibits both necroticand apoptotic deaths of T84 cells. The anti-necrotic effect of quercetin may be attributed to its iron chelator activity rather than a direct $H_2O_2$ scavenging capacity and antioxidant. The present study suggests that quercetin may play a therapeutic role in the treatment of human gastrointestinal diseases mediated by oxidants.

• 약학회지
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• 제49권1호
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• pp.60-67
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• 2005

Nonclassical aminobenz[cd]indole antifolates 4, 5 and 6, in which the glutamic acid moiety of the classical antifolates is substituted by 2-phenylglycinamide or 3-aminobenzamide, were synthesized and their in vitro antitumor activity was evaluated. The purpose of this substitution is that the lipophilicity is enhanced due to the aromatic ring of the target compounds for the passive transport through lipid membrane of cells while the hydrogen bonding of the amide is retained in the active site of the enzyme, thymidylate synthase, where the glutamate is originally present. The target compounds were highly cytotoxic against tumor cell lines of murine and human origin with micromolar to nanomolar $IC_{50}$ values. Most effective was compound 4 ($N^6-methyl-N^6$-[4-[(${\alpha}$(S)-aminocarbonylbenzyl) aminocarbonyl]benzyl]-2,6-diaminobenz[cd]indole)with $IC_{50}$ of 2 nM against SW480, human colon adenocarcinoma cell line, which is 650-fold more potent than the reference compound 3.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.221-224
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• 2001

Effect of hyperosmotic pressure on growth of recombinant Chinese hamster 。 vary cells and Erythropoietin (EPO) production was investigated. Cells were cultivated in batch modes at various osmolalities. When the osmolality increased from 314 to 463mOsm/Kg, specific EPO productivity (qp) was increased up to 1.6-fold but cell growth was inhibited. EPO has a complex oligosaccharide structure that plays an important role in biological activity in vivo. To investigate the influence of hypoerosmotic pressure on the glycosylation, structural analysis of oligosaccharide was calTied out. Recombinant human EPO was produced by CHO cells grown under various osmotic pressure and purified from culture supernatants by heparin-sepharose affinity column and immunoaffinity column. N-linked oligosaccharides were released enzymatically and isolated by paper chromatography. The isolated oligosaccharides were labeled with fluorescent dye, 2-aminobenzamide and analyzed with MonoQ anion exchange chromatography and GlycosepN amide chromatography for the assignment of GU (glucose unit) value. Glycan analysis by HPLC showed that neutral (asialo) oligosaccharide was increased slightly with an increase in osmolality. In portion of sialylated glycan, total relative amount of mono- and di-sialyated glycan was increased but that of tri- and tetra-sialylated glycan decreased as osmolality was increased.

• Radiation Oncology Journal
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• 제12권1호
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• pp.1-8
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• 1994

미량의 변이 유발소에 노출된 세포는 그 다음에 투여되는 다량의 변이 유발소에 대하여 내성을 갖게 된다. 이같은 적응 반응(adaptive response)은 대장균에서 처음으로 밝혀진 이후 주로 인체 말초 혈액 임파구에서 연구가 진행되어 왔다. 그러나 적응 반응이 세포 종류를 막론하고 존재하는 일반적인 현상인지에 대하여, 또한 세포의 고유한 방사선 감수성과의 관계나 그 기전 등에 대하여도 규명되어야 할 필요가 있다. 본 연구는 이같은 의문에 보다 접근하기 위하여 방사선에 매우 민감한 ataxia telangiectasia hemozygote, ataxia telangiectasia heterozygote, 그리고 정상인에서 유래한 인체 임파양세포주를 대상으로 1 cGy의 감마선을 조사하고 일정 시간이 지난 후 다시 50 cGy의 감마선을 조사하여, 감마선에 의해 유도되는 염색체 손상을 측정하였다. 그 결과 1 cGy 전처치시 그 다음 50 cGy에 의한 염색체 손상이 50 cGy 단독 대조군에 비하여 의의 있게 감소하여 적응 반응이 존재함을 알 수 있었다. 세가지 세포주의 방사선 감수성이 각기 달랐으나 적응 반응의 표현 양상은 이와 무관하게 유사하였다. 또한 염색체 손상의 복구에 필수적인 poly(ADP-ribose) polymerase를 억제하는 3-aminobenzamide를 50 cGy 직후에 투여한 실험에서는 적응 반응이 완전히 소실됨을 관찰하여 적응 반응이 어떤 종류의 방사선 손상의 복구 기전과 관계 있음을 추측케하였다. 따라서 임파양세포에서도 적응 반응이 존재함을 본 연구를 통하여 최초로 알 수 있었다. 이는 세포의 방사선 감수성과는 무관한 것으로 나타났으며 그 기전에 있어서는 아직 잘 알려지지 않은 손상 복구 기전을 유도하는 것으로 생각된다.반림프절 비대가 국소치유율, 무병생존율, 생존율에 영향을 미치는 인자이었고 이들 세가지 인자와 전산화단층촬영상의 대동맥림프절 비대가 생존율에 영향을 미치는 인자이었다. 조사선량이 많았으나 치료실패율이 현저하게 높았다. 따라서 이들 환자들을 효과적으로 치료하기 위하여 단순히 치료선량을 증가시키는 것보다 다른 보조적치료, 즉 약물치료와의 병용치료 등이 절실히 요구되며, 또한 국소재발의 억제가 원격전이를 감소시킬 수 있는 한 방법일 수 있다. 따라서 자궁경부암 환자의 방사선 치료에 가장 적합한 방법은 외부조사와 강내 치료를 병행하는 것이나 강내 치료가 불가능한 환자군에 대해서는 적극적 수술방법의 도입을 고려해야하겠다. 합병증의 증가가 관찰되지 않는 점으로 보아 본 연구를 계속 진행함으로써 더 좋은 결과를 얻을 수 있을 것으로 기대된다.기 혈색소 수치 (p<0.0001), 강내 조사(p<0.0004)였고, 조직학적 소견(p<0.29), 유도 화학요법과의 병행치료(p<0.87)는 통계학적으로 유의하지 않았다..0093{\pm}0.0047)\;D^2+(13.31{\pm}7.309$) 였었다. 감마선에 대한 중성자선의 상대적 생물학적 효과비 (RBE)는 y=aD+$bD^2$+c를 다음과 같은 식으로 변형시켜 계산하였다. $$\frac{[-a{pm}\sqrt{a^2-4b\;(c-y}}]}{2{\times}6}$$ 미세핵 발생빈도가 세포당 0.05와 0.8사이에서의 중성자선의 상대적 생물학적 효과비는 $2.37{\pm}0.17$ 이었다. 이상의 결과를 종합하여 볼 때 선량에 따른 미세핵 발생빈도는 기존의 방사선 감수성 test의 결과와 대동소이하여, 앞으로 방사선 감수성을 측정하는 방법으로 이용할 수 있으며, 또한

• Bulletin of the Korean Chemical Society
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• 제33권8호
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• pp.2724-2730
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• 2012

A new heterogeneous acidic catalyst was successfully prepared by impregnation of silica (Aerosil 300) by an acidic ionic liquid, named 1-(4-sulfonic acid)butylpyridinium hydrogen sulfate [$PYC_4SO_3H$][$HSO_4$], and characterized using FT-IR spectroscopy, the $N_2$ adsorption/desorption analysis (BET), thermal analysis (TG/DTG), and X-ray diffraction (XRD) techniques. The amount of loaded acidic ionic liquid on Aerosil 300 support was determined by acid-base titration. This new solid acidic supported heterogeneous catalyst exhibits excellent activity in the synthesis of 2-aryl-2,3-dihydroquinazolin-4(1H)-ones by cyclocondensation reaction of 2-aminobenzamide with aromatic aldehydes under solvent-free conditions and the desired products were obtained in very short reaction times with high yields. This catalyst has the advantages of an easy catalyst separation from the reaction medium and lower problems of corrosion. Recycling of the catalyst and avoidance of using harmful organic solvent are other advantages of this simple procedure.

• Carbon letters
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• 제17권1호
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• pp.45-52
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• 2016

The attachment of 2-aminobenzamide to carboxylated multi-wall carbon nanotubes (MWCNTs)-COOH was achieved through the formation of amide bonds. Then, the functionalized MWCNTs, MWCNT-amide, were treated by phosphoryl chloride to produce MWCNT-quin. The products were characterized by Fourier transform infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, thermogravimetric analysis, derivative thermogravimetric, steady-state fluorescence spectroscopy, and solubility testing. MWCNT-quin showed photo-electronic properties, which is due to the attachment of the 4-hydroxyquinazoline groups to them as proved by steady-state fluorescence spectroscopy. This suggests intramolecular interactions between the tubes and the attached 4-hydroxyquinazoline. The toxicity of the samples was evaluated in human embryonic kidney HEK293 and human breast cancer SKBR3 cell lines, and the viable cell numbers were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) after the cells were cultured for 24 h. Cellular investigations showed that the modified MWCNTs, particularly MWCNT-quin, have considerably significant toxic impact on SKBR3 as compared to HEK293 at the concentration of 5 µg/mL.