• Journal of Pharmacopuncture
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• v.12 no.3
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• pp.25-30
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• 2009

Objectives: To find the potential acupuncture points by using Trypan blue staining on the skin of rat and Nude mouse. Methods: 0.4% Trypan blue was applied to the skin of rat or Nude mouse previously treated by surfactant. Washing by warm saline was followed after enough application of trypan blue and surfactant. Frequency of Trypan blue application should be varied to the experimental animals' condition for visualizing significant spots. Results: Blue spots appeared roughly in symmetry along kidney meridian or stomach meridian. Several spots outside of kidney or stomach meridian were also observed; however, the detail stereoscopic images of those blue spots were slightly different according to the position blue-colored. DiI signals were visualized along blood vessel after DiI injection into the Trypan blue-visualized blue spots. Conclusion: Our method to visualize the potential acupuncture points as a blue spot on rat and Nude mouse skins may contribute to the next step for finding specific flowing channels among blue spots.

• The Korean Journal of Zoology
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• v.12 no.3
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• pp.69-76
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• 1969

The phagocytic petencies and birefringences of peritoneal mast cells of rats treated with particular dyes (neutral red, toluidine blue, pyronin, methylene blue, alcian blue, trypan blue, carmine, orange G, aniline blue, Janus green B, and India ink) were analyzed by means of phase contrast microscopy and polarizing microscopy. In addition, cytomorphic effects of the dyes on the peritoneal mast cells were also discussed. Phagocytic activities or ingestion of the dye particles were not observed in most cases, except for the India ink group. Hardly a macrophage appeared without some dark particles which were ingested or phagocytosed. Trypan blue and aniline blue produced very weak birefringence in the cytoplasm of mast cells but the rest did not produce even in the acid medium (neutral red and toluidine blue). The short and slender ectoplasmic processes of the mast cells and the leucocytes were also found in certain groups. The cytomorphic effects of the dyes on the mast cell were slight and variable.

• Journal of Embryo Transfer
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• v.15 no.1
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• pp.103-108
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• 2000

The object of this study was to find simple and effective methods for the speculation of vitality and scorsome status of bovine spermatozoa. The eosin-nigrosin staining, trypan blue staining, and naphthol yellow S-erythrosin B staining was ofter used for the speculation of vitality and/or acrosome status of bovine spermatozoa, respectively. This study has shown that the combined trypan blue-naphthol yellow S-erythrosin B staining is more accurate and effective for the examination of acrosome status and vitality of bovine spermatozoa.

• Toxicological Research
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• v.5 no.1
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• pp.27-35
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• 1989

When acetaminophen (25mM) was introduced into the perfused rat liver, the hepatic O2 uptake was rapidly inhibited first and then later slow-down. The rapid inhibition was found to be due to mitochondrial blockade, whereas the so-called slow inhibition" was associated with microcirulatory aberrations as evidenced by inhomogneous staining of the liver tissue by trypan blue infusion (0.1%). NaCN (0.5mM) also caused rapid and slow respiratory inhibitions, giving heterogeneous trypan blue staining.ning.

• Journal of Animal Science and Technology
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• v.63 no.1
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• pp.58-68
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• 2021

Several herbs including Artemisia are known to possess conceptive property. In the present study, mouse spermatozoa were incubated with ethanol extract of Artemisia vulgaris leaves. The effect of extract on acrosome exocytosis was studied by labeling spermatozoa with fluorescein isothiocyanate (FITC) peanut agglutinin and by staining with Coomassie blue. Viability and membrane integrity were studied by Trypan-blue staining and hypo-osmotic swelling test. Artemisia extract at very low concentration caused precocious acrosome reaction and loss of sperm viability. Acrosome reaction increased remarkably from 22.63% to 88.42% with increasing extract concentration from 0 to 2,000 ㎍/mL. However, the viability loss of spermatozoa was increased from 11.71% in control to 63.73% in samples treated, evaluated by Trypan-blue staining method. Membrane damage caused by the extract, evaluated by hypo-osmotic swelling test was even low, ranging from 2.27% to only 24.23%. These results indicate that Artemisia extract might block fertilization by causing precocious acrosome exocytosis in spermatozoa. A direct contraceptive effect was tested by injecting the plant extract into the vagina of female mice and then allowing them to mate with normal males. The treated female mice delivered significantly fewer litters in comparison to the control.

• Korean Journal of Agricultural and Forest Meteorology
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• v.14 no.4
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• pp.246-253
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• 2012

Despite frequent freezing injury to tea trees due low temperature, drought, and strong wind during wintertime, no comprehensive measurements have been taken. We selected and examined 9 locations in Hwagae-myeon and 4 places in Agyang-myeon, Hadong-gun, Gyeonsanggnam-do where low temperature damage had occurred between December 2010 and February 2011. Our objective is to examine the effect of frost damage on the morphological symptom and harvest of a tea tree exposed to a constant low temperature environment during wintertime. The results of our analyses on meteorological environment, tea leaf chromaticity, water content and trypan blue are as follows: (1) the number of days with temperature of $-10^{\circ}C$ or less, which were subject to frost damage to a tea tree were 8 and 13.6% during the winterization period in 2011; (2) the accumulated time was 1,308 minutes, and the longest duration at $-10^{\circ}C$ was 588 minutes from 21:08 p.m. 15 January to 7:30 a.m. $16^{th}$ January. The rainfall was only 104 mm which was 306 mm less than the previous year; (3) the lightness L values in 2011 were higher than in 2012 due to dehydration and necrosis by blue discoloration and red discoloration at all areas in chromaticity measurement; (4) the water content in a tea leaf in 2011 was higher than in 2012 due to low rainfall and strong wind, and almost no cell death phenomenon was observed from normal tea leaves subject to no low temperature stress in a trypan blue analysis; and (5) partial coloration due to cell death, however, took place in the leaves damaged by blue discoloration subject to low temperature stress, and most coloration due to cell death took place in the leaves damaged by red discoloration.

• Journal of the korean veterinary medical association
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• v.16 no.1
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• pp.25-28
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• 1980

The studies were undertaken with the objective to compare six staining techniques, nigrosineosin, eosin-nigrosin, trypan blue, Congo red-nigrosin, fast green-erythrocin and fast green-eosin and select one of these staining techniques to measure live-dead

• Radiation Oncology Journal
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• v.24 no.3
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• pp.179-184
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• 2006

[ $\underline{Purpose}$ ]: To explore the effect of recombinant human EGF on the proliferation and survival of human fibroblast cell lines following irradiation. $\underline{Materials\;and\;Methods}$: Fibroblast was originated human skin and primary cultured. The trypan blue stain assay and MTT assay were used to study the proliferative effects of EGF on human fibroblast cell lines in vitro. An incubation of fibroblasts with rhEGF for 24 hours immediately after irradiation was counted everyday. Cell cycle distributions were analyzed by FACS analysis. $\underline{Results}$: Number of fibroblast was significantly more increased rhEGF (1.0 nM, 10 nM, 100 nM, 1,000 nM) treated cell than control after 8 Gy irradiation. Most effective dose of rhEGF was at 160 nM. These survival differences were maintained at 1 week later. Proportion of S phase was significantly increased on rhEGF treated cells. $\underline{Conclusion}$: rhEGF cause increased fibroblast proliferation following irradiation. We expect that rhEGF was effective for radiation induced wound healing.

• Journal of the East Asian Society of Dietary Life
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• v.24 no.6
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• pp.739-747
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• 2014

The aim of the study was to determine the effects of Ixeris dentata extract (IDE) on anticancer activity in human breast cancer MDA-MB-231 cells at both cellular and molecular levels. The cells were cultured in the presence of 0, 20, 30 and $40{\mu}g/mL$ Ixeris dentata extract for 24 hours, respectively. At the end of culture, cytochemical analyses for MTT activity, trypan blue dye exclusion, Annexin V-FITC Apoptosis, and radical oxygen species (ROS) were conducted. RT-PCR was also performed to determine whether or not alterations in cell viability affect the Bax/Bcl-2 ratio. MTT assay showed that relative cell viability decreased in a dose-dependent manner (p<0.05). Reduction of cell viability matched well with increased cell membrane permeability as determined by trypan blue dye exclusion test (p<0.05). The rates of intracellular ROS also increased in a similar manner to those of TB-stained cells. There was an associated shift of apoptotic cells from early to late apoptosis between the 30 and $40{\mu}g/mL$. Bax/Bcl-2 ratio significantly increased along with significant decreases in Bcl-2 expression between 30 and $40{\mu}g/mL$ groups (p<0.05). In conclusion, anticancer activity of Ixeris dentata extract is modulated by a reduction in cell viability along with increased membrane permeability, leading to ROS accumulation within cells, and subsequently cell death through an apoptotic pathway that involves Bax and Bcl-2 in human breast cancer MDA-MB-231 cells.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.475-490
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• 2005

본 실험은 흡연이 치주인대세포의 기능에 미치는 영향을 알아보기 위해 담배 부산물 중 하나인 니코틴이 지주인대세포의 부착과 성장 및 세포의 가역성에 미치는 영향을 관찰하였다. 니코틴이 치주인대세포의 부착에 미치는 영향을 관찰하기 위하여 치주인대세포에 니코틴을 투여한 후 1, 3, 6, 12, 24시간째 trypan blue 염색 후 부착된 세포수의 측정과 H&E stain후 형태학적 관찰을 하였고 니코틴이 치주인대세포의 성장에 미치는 영향을 관찰하기 위하여 치주인대세포에 니코틴을 투여한 후 1, 4, 7, 11, 14일째 trypan blue 염색 후 증식된 세포수의 측정과 H&E stain후 형태학적 관찰을 하였다. 또 니코틴의 세포독성효과의 가역성 평가를 위하여 니코틴을 투여하고 1, 4, 7, 11일째 니코틴이 없는 새로운 배지로 갈아주어 2일 더 배양한 후 trypan blue로 염색하여 세포수를 측정, 비교하였고 H&E 염색 후 형태학적 관찰을 하였다. 실험결과 니코틴 농도가 증가함에 따라 치주인대세포의 부착율은 감소되었고 24시간 배양 후 2mg/ml, 0.5mg/ml, 0.1mg/ml니코틴 투여군에서 통계학적으로 유의성 있는 부착억제가 관찰되었고(P<0.01) 형태학적 관찰결과 2mg/ml, 0.5mg/ml 니코틴 투여군에서는 대조해서 관찰되는 세포질의 확장이 관찰되지 않았다. 니코틴이 치주인대세포의 증식에 미치는 효과를 살펴본 결과 2mg/ml, 0.5mg/ml, 0.1mg/ml 니코틴 투여군에서 증식억제가 관찰되었고(P<0.01) 0.005mg/ml이하의 니코틴 투여군에서는 아무런 변화도 관찰되지 않았다. 14일 배양 후 형태학적 관찰결과 0.1mg/ml 니코틴 투여군에서 세포질내 공포를 관찰할 수 있었고 2mg/ml, 0.5mg/ml 니코틴 투여군에서는 세포의 괴사가 나타났다. 니코틴의 세포독성효과의 가역성평가결과 2mg/ml이상의 니코틴은 세포에 비가역적인 손상을 일으키며 0.5mg/ml, 0.1mg/ml 니코틴에 의한 세포독성은 초기에는 가역적이나 장기간 세포에 노출시키면 비가역적인 손상을 야기하는 것으로 나타났다.(P<0.05) 본 실험결과 담배의 구성성분 중 니코틴은 농도 의존적으로 치주인대세포의 부착과 증식에 영향을 주었고 니코틴의 치주인대세포에 대한 독성효과는 농도가 증가할수록, 시간이 지닐수록 비가역적으로 나타났다. 따라서 흡연은 치주조직재생에 영향을 미칠 것으로 생각되며 흡연의 기간과 정도가 치주질환의 심도 및 치주치료 후 불량한 치유반응과 관계가 있을 것으로 사료된다.