• 제목/요약/키워드: Crystal violet

검색결과 157건 처리시간 0.03초

Anisakis 형(型) 유충(幼蟲)에 관한 연구(硏究) (Studies on Anisakis Type Larvae)

  • 임정택
    • 대한수의학회지
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    • 제15권2호
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    • pp.293-307
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    • 1975
  • As it has been known recently that anisakis type larvae harbouring in marine fishes are a causal agent of zoonosis to human and probably to land living mammal animals, attention was focused on the study on the larvae in an aspect of epidemiology or epizootiology. The present work was conducted from 1966 to 1975 for i) survey on the harbouring status of anisakis type larvae in marine fishes of this country, ii) observation on the response to the experimental infestation of the larvae to the pigs, in the reason that they could well fetid raw fish viscera occasionally containing the larvae as a high protein source of swine food, and iii) observation on the larval resistance and response to vermicidal agents for the purpose of prevention of the larval infection to the mammal animals. The data obtained in the studies were summarized as follows: 1. In the survey on the status of larvae harbouring in main species of marine fishes of this country, 15 species, a total of 1,940 fishes, were observed and the result was summarized in table 2. Average number of larvae, in upper rank of 5 out of all 15 species of fishes, were as highest as 156 larvae ranging 74 to 450 in Pseudosciaena manchurica (chamjogi), 54.5 ranging 15 to 240 in Trichiurus haumela (kalchi), 35.6 ranging 8 to 112 in Trachurus japonica (junggengi), 30.6 ranging 4 to 65 in Parapristipama trilineatum (benjari) and 20.5 ranging 3 to 48 in Nibea argentata (boguchi) respectively. In morphological observation, size of the larvae in the fishes were varied, ranging from 2 to 32mm long, and a tendency to larger size and number of larvae in the fishes, which were wider sea migration, higher age and lager bodily size, was observed The favorite places harbouring the larvae in fishes were mainly around the intraperitoneal viscera such as mesentery, omentum, liver, pyloric suspensory, fat tissue and cloaca, and rarely in body muscles of fish. Fishes heartily infested with the larvae showed stunted growth decreased egg formation and severe damage of liver. 2. In the experimental infestation of the larvae to normal pigs, as illustrated in table 3, a group with large dose of larvae (a total of 1,800 larvae, 300 larvae Per dose, twice in a dart for 3 days) showed acute clinical syndrome terminatine death with a week course, whereas two groups with less dose of larvae (a total of 180~360 larvae, 10 larvae per dose, at 5 days interval for 70~180 days) showed subclinical syndrome with remarkably stunted growth as. much as approximately one half of body size in contest to the control pigs. In the pathological findings, a group with large dose of larvae showed macroscopically larvae penetrating to the gastric wall with severe gastroenteritis, and histopathologically various acute lesions caused by active larvae penetration into the wall of stomach and interstine, whereas two groups with less dose of larvae showed chronic lesions such as hypertrophy and verminous granulomatous swelling of gastric wall, suggesting strongly the possibility of natural infestation of larvae to swine. 3. In the resistance of the larvae to the chemical solutions, the larvae tolerated for 2 days in 15 percent solution of sodium chloride and acetic acid, and for 7 days in 70 percent solution of ethyl alcohol. In the resistance to the temperature, the larvae died within 1 second at $62^{\circ}C$ and tolerated for 24 hours at $-3^{\circ}C$, 12 hours $-5^{\circ}C$ respectively. 4. For the experiment on the vermicidal effect to larvae, general vermicidal drugs such as Neguvon, Combantrin, antimony Potassium, piperazine adipate and piperazine dihydrochloride, oxidizer such as potassium permanganate and potassium chlorate, and dyes such as gentian violet and crystal violet were used, and among them, as illustrated in table 6, potassium permanganate was proved as the best. In the successive test for the practical use of potassium permanganate, vermicidal effect in seawater solution of potassium permanganate and common-water solution of potassium permanganate were compared, and then retested by dipping the fish viscera including the larvae into the two different solutions of potassium permanganate. The result through these tests indicated that 0.01 percent common water and sea-water solution of potassium permanganate could be apparently recommended as a preventive vermicidal solution, having 90 to 100 percent vermicidal effect by dipping for 12 to 24 hours even though sea-water solution of potassium permanganate had a tendency to slightly less effect than the common-water solution of potassium permanganate (Table 8).

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음용 차의 항우식 효과 (Anticariogenic Effects of Different Types of Tea)

  • 정일용;이주영;김미아;이대우;양연미;김재곤
    • 대한소아치과학회지
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    • 제44권4호
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    • pp.437-445
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    • 2017
  • 이 연구의 목적은 한국인들이 일상적으로 마시는 서로 다른 네 종류의 차 티백을 이용하여 차가운 생수나 뜨거운 생수로 5분 또는 10분 동안 우려내는 일상적인 방법으로 차를 추출하였을 때 각각의 추출물이 Streptococcus mutans 세균막 성장에 미치는 효과를 알아보고자 하였다. 균주는 S. mutans UA 159를 이용하였고 1%의 sucrose와 각각의 차 추출물을 웰 플레이트에 넣고 배양하였다. 추출 온도에 따른 세균막 형성을 비교 했을 때 녹차와 홍차에서 추출 온도가 높을 때 세균막 형성이 적었고 통계적으로 유의하였다(p < 0.05). 추출 시간을 달리 하고 $72^{\circ}C$ 온수로 많은 양을 추출 했을 경우 세균막 형성을 비교 했을 때는 네 종류 차 모두 추출 시간에 대해서 통계적으로 유의하지 않았다. 한편 추출량에 따른 비교에서는 녹차와 홍차의 경우 같은 시간, 같은 온도로 추출하였을 때 추출량이 적다면 오히려 세균막 성장이 증가하였다.

참외 추출물의 Quinone Reductase 유도활성 및 간암세포 증식 억제효과 (Quinone Reductase Inductive Activity and Growth Inhibitory Effect against Hepatoma Cell of Oriental Melon Extract)

  • 김혜숙;구강모;서전규;강영화
    • 생물환경조절학회지
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    • 제18권4호
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    • pp.448-453
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    • 2009
  • 본 연구에서는 참외 추출물의 항암활성에 대해 알아보기 위해 참외를 부위별로 나누어 quinone reductase 유도활성과 다양한 간암세포에서의 증식 억제활성을 조사하였다. 참외 꼭지와 참외 줄기 잎 부위에서 농도의존적으로 QR 유도활성이 증가하였고, $200{\mu}g/mL$ 농도에서는 각각 3.9, 1.5배의 유도활성을 나타내었다. 암세포 사멸 활성 측정법을 통한 항암활성 평가 실험에서 마우스 유래의 간암세포인 Hepa1c1c7 세포에 대해서 조사한 결과 꼭지와 줄기 잎 부위에서 높은 암세포 독성을 보였다. 이러한 결과를 기초로 인체유래의 암세포에 대한 항암활성을 평가하기 위해 인체유래 간암 세포주인 HepG2에 대한 세포 증식 억제활성을 농도별로 조사하였다. 꼭지와 줄기 잎 부위 모두 인체유래 간암 세포에 대해 증식 억제효과를 보여주었지만, 특히 꼭지 부위는 최고농도에서 60.3%의 높은 증식 억제효과를 보였다. 그러나 마우스 유래의 간암세포에 대한 활성보다 인체유래 간암세포에 대한 활성이 낮게 나타났다. 참외의 꼭지 추출물에서 QR 유도활성과 항암활성을 확인함으로써 향후 참외 비가식 부위의 기능성 소재로의 이용화에 대한 연구가 필요할 것으로 생각된다.

팔진탕합화적환(八珍湯合化積丸)과 Adriamycin의 병용처리시 나타나는 synergistic 항종양(抗腫瘍) 효과(效果)에 관(關)한 작용기전 연구(硏究) (Study on Synergistic Anti-tumor Effect of Combination with Adriamycin and Palginhonhapwhajucwhan)

  • 문구;문석재;원진희;조정연;박상구;송봉길;박래길;이병구
    • 대한한방내과학회지
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    • 제21권3호
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    • pp.443-452
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    • 2000
  • Objective : This study was designed to evaluate the synergistic effect on cytotoxicity of combination with adriamycin and Palginhonhapwhajucwhan, a traditional prescription for cancer treatment in oriental medicine, in Chang, HL-60, Hep-3B and Alexander cells. Methods : We observed cell viability in Chang, HL-60, Hep-3B, and Alexander cells by crystal violet staining. Those cells were treated with various concentrations of adriamycin alone, Palginhonhapwhajucwhan alone and combination of two medications for 10 hr. On condition of $0.5{\mu}l/ml$ adriamycin alone, $15.6{\mu}l/ml$ Paljintanghapwhajucwhan alone and combination of two medications, at first, we observed colony forming of Chang and HL-60 cells. Second, we observed DNA fragmentation by agarose electrophoresis in Chang, HL-60, Hep-38 and Alexander cells. Third, we measured the catalytic activation of caspase-1, 2, 3, 6, 8, and 9 protease in Chang cells and caspase-3 protease in Chang, HL-60, Hep-3B and Alexander cells by using fluorogenic substrate. Finally, we isolated mRNA of Fas in Chang, HL-60, Hep-38 and Alexander cells and observed that Fas gene was amplified by RT-PCR Results : 1. The combination of adriamycin and Palginhonhapwhajucwhan synergistically augmented the cytotoxicity of Chang and HL-60 cells whereas did not in Hep-38 and Alexander cells. 2. Cotreatment of two drugs also markedly inhibited the colony forming ability both in Chang and HL-60 cells. 3. The cytotoxicity of these medicatons was revealed as apoptosis characterized by high molecular wight DNA fragmentaton. 4. The apoptotic cytotoxicity was mediated by activation of caspase-3 protease in Chang cells. 5. Synergistic increase in apoptotic cytotoxicity by combination of two medications was dependent on the expression of Fas in cancer cells. Conclusions : Combination of adriamycin and Palginhonhapwhajucwhan significantly augmented apoptotic cytotoxicity of Fas-positive cells such as Chang and HL-60 cells via acticaton of apoptosis signaling pathway.

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광화학적 반응을 이용한 편극 패턴된 강유전체 표면에 금속 나노입자의 증착에 관한 연구 (Growth of Metal Nano-Particles on Polarity Patterned Ferroelectrics by Photochemical Reaction)

  • 박영식;김정훈;양우철
    • 한국진공학회지
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    • 제20권4호
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    • pp.300-306
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    • 2011
  • 본 연구는 편극 패턴된 강유전체 단결정 $LiNbO_3$ (0001) 기판에 광화학적 환원반응을 이용하여 금속(Au, Ag) 나노입자를 증착시키고, 금속 입자의 종류와 표면의 극성에 따른 나노입자의 표면 분포를 원자간력현미경(AFM)으로 조사하였다. 전극 인가에 의해 주기적으로 편극 패턴된 강유전체 단결정 $LiNbO_3$ (periodically polarity-patterned $LiNbO_3$: PPLN)을 기판으로 사용하였으며, PPLN의 각 영역의 편극 방향은 Piezoresponse force microscopy로 확인하였다. 금속(Ag, Au) 나노 입자는 금속이 포함된 수용액에 PPLN 기판을 넣고, 자외선 램프로 30초에서 3분간 노출시켜 광환원 반응으로 기판에 증착시켰다. 시료 성장후, 공기 중에서 AFM을 이용하여 나노입자의 형태, 크기, 및 표면분포를 조사하였다. Ag 입자의 경우, -Z 편극 영역보다 +Z 편극 영역에 크고 밀도가 높은 나노 입자가 증착되었으며, 특히 편극 경계 부분에 가장 큰 Ag 나노입자가 증착되어, 나노선 모양으로 성장됨이 확인되었다. 그러나 Au 입자의 경우는 편극 경계부분에 입자가 증착되는 경향이 없었다. 두 입자 모두 자외선 노출시간이 증가함에 따라, 증착된 나노입자의 크기는 증가하는 경향을 보였다. 이와 같이 증착된 금속 나노입자가 강유전체의 표면편극에 따라 다른 분포로 성장되는 것을 강유전체 표면 극성에 따른 표면 밴드구조 변화, 광전 효과 및 표면의 전기장의 불균일성에 의한 수용액 속의 금속 양이온과 자외선에 의해 생성된 전자와의 광화학적 반응에 대한 모델로 논의할 것이다.

Fibronectin type III 10 도메인이 조골양 세포에 미치는 생물학적 영향 (Biological Effects of Fibronectin Type III 10 domain on Human Osteoblast-like cells)

  • 이창석;장준혁;김태일;이용무;류인철;정종평;한수부;구영
    • Journal of Periodontal and Implant Science
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    • 제34권2호
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    • pp.293-301
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    • 2004
  • 1. 연구 목적 Fibronectin은 세포외기질의 주요성분인 거대 당단백질로서, 조골세포의 부착과 증식 및 이동능에 중요한 역할을 담당한다고 알려져 있다. 이러한 fibronectin의 조골세포에 대한 영향을 실제 임상에 적용하기 위해서, 전체 fibronectin 단백질을 사용하는 것은 면역학적으로나 경제적으로 많은 단점을 안고 있어서, 유효한 반응단위만을 추출하여 활용하는 것이 바람직한 방법으로 알려져 있다. 이 연구의 목적은 세포부착에 주로 관여하는 fibronectin type III분절 중 10번 도메인이 조골양 세포에 미치는 영향을 전체 fibronectin단백질과 fibronectin type III 7-10 도메인 분절과 비교, 관찰하는 것이다. 2. 연구 방법 사람의 fibronectin을 기초로 한 적절한 primer로서, 유전자 재조합법을 이용하여 fibronectin type III 10 도메인과 fibronectin type III 7-10 도메인 분절을 얻었으며, 전체 fibronectin분자는 상용으로 준비하여 24-well 세포배양 용기에 도포하였다. 배양된 조골양세포(HOS cell)를 $1x10^5$ cells/well의 농도로 각 well에 분주하여 $37^{\circ}C$에서 1시간 배양을 하였다. Cell adhesion assay를 실시하기 위해 10% formaldehyde로 고정시키고 1% Crystal Violet으로 염색하여 광학현미경을 관찰 후 2% SDS를 처리하여 microplate reader기를 이용하여 570nm에서 혼탁정도를 측정하였다. 음성대조군으로는 RPMI 용액을 사용하였다. 동일한 방법을 이용하여 준비된 $35mm^2배양접시에 HOS cell을 $37^{\circ}C$에서 4일간 배양 후, MTS assay를 이용하여 세포 증식도에 미치는 영향을 관찰하였다. 6일째 405nm에서 활성화된 세포에서 분비된 p-nitrophenol을 이용한 alkaline phosphatase activity를 측정하였다. 3. 결과 및 고찰 Fibronectin type III 10 도메인은 HOS cell에 대한 생물학적인 효과면에서, 전체 fibronectin 분자 및 fibronectin type III 7-10 분절과 통계적으로 유사한 세포부착도를 보여주었으며, 세포증식도와 alkaline phosphatse 활성도면에서도 큰 차이가 나타나지 않았다. 이상의 연구결과로 볼 때, fibronectin type III 10 도메인이 조골세포의 증식을 목적으로 사용하는 생체재료의 표면개질 부착물질로 응용할 수 있는 가능성이 있다고 하겠다.

진무탕(眞武湯)이 $H_2O_2$로 유도된 C6 Glial 세포사에 미치는 영향 (Protective Effect of Jinmu-tang on $H_2O_2$-induced Cell Death in C6 Glial Cells)

  • 최정훈;신용진;하예진;조문영;유주연;이숭인;신선호
    • 대한한방내과학회지
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    • 제33권3호
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    • pp.272-283
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    • 2012
  • Objectives : The purpose of this study was to investigate the mechanism of protective effect of Jinmu-tang (JMT, Zhenwu-tang) extract on $H_2O_2$-induced cell death in C6 glial cells. Methods : Cultured C6 glial cells of white mice were pretreated with JMT extract and exposed to $H_2O_2$ for inducing cell death. We measure the cell viability by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and investigate the cell morphology using a light microscope after crystal violet (CV) staining. Reactive oxygen species (ROS) formation was analyzed using a flow cytometer and a fluorescent microscope after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). DNA fragmentation was analyzed using a flow cytometer after propidium iodide (PI) staining and nuclei morphology was investigated using a fluorescent microscope after 2-[4-amidinophenyl]-6-indo-lecarbamidine dihydrochloride (DAPI) staining. We analyzed expression of Bax, processing of procaspase-3 and poly (ADP-ribose) polymerase (PARP), and activation of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) by western blot method. Tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) secretion was analyzed using Quantikine kit. Results : We determined the elevated cell viability by JMT extract on $H_2O_2$-induced C6 glial cell death. ROS formation, DNA fragmentation, $I{\kappa}B{\alpha}$ phosphorylation, NF-${\kappa}B$ activation, and secretion of TNF-${\alpha}$ induced by $H_2O_2$ are inhibited by JMT extract pre-treatment. JMT extract inhibits Bax expression, processing of caspase-3 and PARP that are critical biochemical markers of apoptotic cell death. Conclusions : These results suggest that JMT extract has a protective effect on $H_2O_2$-induced C6 glial cell death in various pathways.

팔물탕합화적환(八珍湯合化積丸)의 항종양(抗腫瘍) 효과(效果)에 관(關)한 연구(硏究) (Experimental Studies on Antitumor Effects of Paljin-tang hab Hwajuck-hwan)

  • 송봉길;이건업;원진희;문구;문석재;소홍섭;박래길;김성진
    • 대한한방내과학회지
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    • 제21권1호
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    • pp.65-73
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    • 2000
  • Objectives : The effects of cotreatment of adriamycin and ethanol extract of herb (Palgin-tang hab Hwajuck-hwan a traditional medicine for cancer treatment in oriental medicine) on the induction of apoptotic cell death were investigated in human liver origin cell lines, Chang. Methods : Chang(ATCC) liver cells were cultured in RPMI-1640(Gibco SRL Co, Gaithersburg, MD) badge including 10% fetal bovine serum. Chang liver cells were treated with various concentrations(from 10 to $0.16{\mu}l$) of adriamycin and herb extract(from 500 to $31.25{\mu}l$) After 48h later, the cells were tested for viability by Crystal violet staining assay. Adriamycin and Herb extract induced ladder pattern of DNA fragmentation in Chang cells. Genomic DNA was isolated and separated on 1.5% agarose gels. The DNA was stained with ethidium bromide and visualized under UV light. Results : The death of Chang cells was synergistically induced by the cotreatment of adriamycin and ethanol extract of herb. In addition, the cotreatment-induced cell death of Chang cells was mediated by apoptotic death signal processes. The phosphotransferase activity of JNK1 remained in a basal level in Chang cells which was treated individually with the adriamycin and ethanol extract of herb. However, it was markedly increased in Chang cells which was cotreated with adriamycin and ethanol extract of herb. In addition, the expression of Fas and FasL was markedly induced by the cotreatment of adriamycin and herb extract. For a while, the expression of Sax was a eminently increased by the ethanol extract of herb. However, Scl2 expression was not affected by the individual or cotreatment of adriamycin and herb extract. Conclusions : our results suggest that the cotreatment of adriamycin aM ethanol extract of herb induces synergistic apoptotis of human liver origin Chang cells via the upregulation of JNK, Fas, FasL and Bax.

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$CoCl_2$로 유도된 C6 신경교세포의 사멸에 대한 억간산(抑肝散)의 보호 효과 (Protective Effects of Ukgan-san in $CoCl_2$-induced Cell Death of C6 Glial Cells)

  • 조문영;신용진;하예진;우찬;김태정;유주연;최용석;최정훈;신선호
    • 대한한방내과학회지
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    • 제34권2호
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    • pp.178-191
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    • 2013
  • Objectives : In this study, we made an effort to investigate the protective mechanism of Ukgan-san (UGS) extracts on hypoxia-induced C6 glial cell death. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were analysed with microscope after staining with crystal violet (CV). Reactive oxygen species (ROS) formation was assessed by flow cytometer after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). We also analyzed expression of hypoxia-inducible factor-1 alpha (HIF-$1{\alpha}$) and p53, processing of procaspase-3 and procyclic acidic repetitive protein (PARP) by western blot method. Results : We estimated the elevated cell viability by UGS extract on $CoCl_2$-induced C6 glial cells. UGS attenuated $CoCl_2$-induced ROS formation in C6 glial cells and also showed a protective activity compared to antioxidants and exhibited abrogation of LDH-released by $CoCl_2$. UGS suppressed the typical apoptotic cell death markers, caspase-3 and PARP activation. UGS inhibited $CoCl_2$-induced HIF-1${\alpha}$ expression which is known as a major regulator for hypoxia-induced cell death, and suppressed p53 expression. Conclusions : These results suggest that UGS extract contains protective constituents for hypoxia-induced C6 glial cell death.

Post HCV Infection Due to MX Gene Stimulation Produced Post Treatment with Imported and Locally Produced Egyptian Biosimilar IFN

  • Mohamed, Shereen H;Mahmoud, Nora F;Mohamed, Aly F;Kotb, Nahla S
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권14호
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    • pp.5635-5641
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    • 2015
  • Background: Cirrhosis is regarded as a possible end stage of many liver diseases, including viral infection. It occurs when healthy liver tissue becomes damaged and is replaced by scar tissue and finally may lead to hepatocellular carcinoma. Interferons (IFNs)are two general categories, type I and II. Type I includes one beta interferon and over 20 different alpha interferons. Alpha interferons are very similar in how they work, interacting with other proteins on cells like receptors. The main objective of this study was to compare Mx gene productivity post different cell line treatment with imported and Egyptian biosimilar locally produced IFNs, as well as the efficacy of those tested IFNs. Also, an assessment was made of sensitivity of different cell lines as alternatives to that recommended for evaluation of antiviral activity. Materials and Methods: Different cell lines (Vero, MDBK and Wish) were employed to evaluate cytotoxicity using the MTT assay. Antiviral activity was evaluated compared with standard IFN against VSV, Indiana strain -156, on tested rh-IFNs (imported; innovated and Egyptian biosimilar locally produced IFNs) in the pre-treated cell lines previously mentioned. The virus was propagated in the Wish cell line as recommended. Finally we estimated up-regulation of the Mx gene as a biomarker. Results: Data recorded revealed that test IFNs were safe in test cell lines. Viability was around 100%. Locally tested interferon did not realize the international potency limits, while the imported one was accepted compared with the standard IFN. These results were the same either using infectivity titer reduction assay or crystal violet staining of residual non- infected cells. Mx protein production was cell type related and confirmed by the detected Mx gene expressed in imported and locally produced IFN pre-treated cell lines. The expression of the gene was arranged in the order of Vero> wish > MDBK for the imported IFN, while for the Egyptian biosimillar locally produced one it was MDBK> Vero> wish. With regard to the antiviral activity there was a significant difference of imported IFN potency compared with the locally produced IFN (P<0.05), the IFN potential (antiviral activity) was not cell line related and showed non-significant difference for each separate product. Conclusions: Vero cells can be used as an alternative cell line for evaluation of IFN potency in case of unavailable USP recommended cell lines. Alternative potency evaluation assay could be used and proved significant difference in IFN potency in case of local and imported agents. Evaluation of antiviral activity could be used in parallel to viral infectivity reduction assay for better accuracy. Mx gene can be used as a marker for IFN potential.