• Korean Journal of Veterinary Research
• /
• v.64 no.1
• /
• pp.2.1-2.8
• /
• 2024

This study was performed to assess the antiapoptotic effect of canine platelet-rich plasma (PRP) treated on the canine adipose-derived mesenchymal stem cells (cMSCs) under cold ischemic conditions. The effect of preventing apoptosis of cMSCs was evaluated in the apoptotic condition induced by cold ischemic injury in vitro. To determine the progression of apoptosis, the changes in cell nucleus were observed using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining. In addition, we examined the mitochondrial membrane potential (MMP) and caspase-3 activity. When the cold hypoxic injury was applied to cMSCs, the apoptotic change was observed by DAPI staining, mitochondrial staining for MMP, and caspase-3 assay. PRP significantly decreased the number of apoptotic cells. Nuclear shrinkage and fragmentation of apoptotic cells in control groups were observed by DAPI staining. The MMP was recovered by the treatment of PRP. In addition, when the luminescence intensity was measured for caspase-3 activity, the value was significantly higher in the PRP treated groups than the control groups. The results of this study showed that the PRP may have a beneficial effect on apoptosis induced by cold ischemic injury.

• Journal of Environmental Science International
• /
• v.18 no.1
• /
• pp.9-14
• /
• 2009

Pfiesteriaand Pfiesteria-like organisms were reported to be linked to major fish kills(involving well over a billion fish) in North Carolina and Maryland estuaries on the U.S. east coast during the 1990s. Occurrences of these species have been recently reported from Korean waters including Chinhae Bay and the coast of Yeosu. In this study, the life cycle of Cryptoperidiniopsis brodyi and Pfiesteria piscicida were examined using DAPI staining. Their excystment and growth were stimulated directly by the addition of prey cells such as Rhodiminas salina. Amoeboid stages in C. brodyi and P. piscicida were never observed in culture, even after addition of filter-sterile fish mucus and tissue. The dominant life cycle stages consisted of motile flagellated zoospores and cysts. A typical dinoflagellate life cycle was demonstrated by direct observation and DAPI staining.

• Korean Journal of Microbiology
• /
• v.37 no.1
• /
• pp.61-65
• /
• 2001

Epifluorescence microscopy was used to observe epiphytic bacteria directly on plant leaf surfaces as well as indirectly in the leaf liberating solution by staining with fluorochromes of 4',6-diamidino-2-phenylindole (DAPI) and acridine orange(AO). Epiphytic bacteria could not be well observed on the leaf surface by staining with AO due to an intrusive orange or red background fluorescence. However, DAPI gave us clear epifluorescent images of the bacteria on the leaf. On the contrary, epiphytic bacteria in the liberating leaf solution were well observed on filters stained by both types of fluorochrome, although DAPI showed better fluorescent images than AO and not necessarily required a washing step of the filters stained. The optimum conditions of the DAPI stains were 5 $\mu$g/ml for 5 min both for leaves and for filters of the liberating solution. It was confirmed that a critical step in the epifluorescence microscopy of leaf surfaces was to minimize release of water from the leaf. For this, the stained leaf samples were put on a filter paper, kept in a dry oven at $70^{\circ}C$ for 2 min instead of air-drying, and then immediately observed by epifluorescence microscopy. The established technique was applied to enumerate epiphytic bacteria on oak tree leaf surfaces.

• Korean Journal of Acupuncture
• /
• v.26 no.1
• /
• pp.79-84
• /
• 2009

Objective : In 1960's Bonghan Kim's team found BongHan(BH) ducts which were presumed as acupuncture meridians and BH corpuscles. They asserted Bonghan theory and SanAl theory which was involved in cell division and cell restoration. However, many other experiments which had been operated to demonstrate and find the existence of BH ducts had failed because of the secret of blue stain drugs. During the last several years, BongHan theory has been revived through experimental researches to find the anatomical structures of BH ducts and corpuscles by Soh's Biomedical Physics Lab. Soh's research team used the staining with Janus Green B, Alcian blue, nanoparticles and Acridine Orange. We used DAPI staining to find the existence of BH ducts and the corpuscles and to observe nuclear arrangement. Methods : We used japan white rabbits as experimental animals. BH ducts and corpuscles were stained with DAPI. The nucleus configuration in BH ducts stained with DAPI were observed with microscope. Results : In this study, we found thread like structures in silver white color distinguished from the blood vessels, nerves and lymph vessels. These thread like vessels in the linear duct shape were connected to same colored mass in the ball shape. Thread like structures we found could be separated easily from the surrounding other organ mass. The nuclei of the thread like structure in DAPI staining, are about 10${\sim}$20${\mu}m$ length, in rod shape and linear arrangement. Conclusion : We concluded that the thread like structure we found was same vessel reported by Soh's research team, BongHan ducts and corpuscle.

• Journal of Life Science
• /
• v.18 no.9
• /
• pp.1312-1315
• /
• 2008

High throughput-drug screening plays a pivotal role for early stage of drug discovery process. In the course of assay development for screening of cell death-inducing agents, a protocol that is simple, time-saving, and high throughput-compatible was designed which was confirmed that the protocol can be worked by a HTS-compatible machine. In 96-well format, PC-3 cancer cells (1${\times}10^{4}$ cells/ml) were cultured for 24 hr. After 24 h-incubation with various medicinal plants extracts, the cells were then stained with DAPI for 30 min. The fluorescence intensity of the stained cells was measured semi-automatically with a multilabel counter. To test whether the present assay system effectively works, we screened about 850 medicinal plant extracts, and selected 1 positive crude extracts that contained cell death-inducing activity. These results suggest that the protocol is highly amenable to HTS implementation for a cell death-inducing agent(s).

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.5
• /
• pp.2123-2128
• /
• 2014

A current hurdle in cancer management is the intrinsic or acquired resistance of cancer cells to chemical agents that restricts the efficacy of therapeutic strategies. Accordingly, there is an increasing desire to discover new natural compounds with selective toxicity to combat malignancies. In present study, the cytotoxic and apoptosis-inducing activities of ferutinin, a terpenoid derivative from Ferula ovina, were investigated on human breast (MCF7) and bladder (TCC) cancer cells as well as normal fibroblasts (HFF3).The toxicity and DNA damage inducing effects of ferutinin were studied by MTT and comet assays, DAPI and PI staining and DNA laddering. The $IC_{50}$ values of ferutinin were identified and compared with routine prescribed drugs, doxorubicin and vincristine, by MTT test. Alkaline comet assay and DAPI staining revealed DNA damage due to ferutinin, which was significantly (p<0.001) higher in MCF7 and TCC than HFF3 cells. Apoptosis induction was evidenced by PI staining and DNA laddering. Our results suggest that ferutinin could be considered as an effective anticancer agent for future in vivo and clinical experiments.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.12
• /
• pp.4897-4902
• /
• 2014

Purpose: To investigate the anticancer effects and underlying mechanisms of parthenolide on HepG2 human hepatocellular carcinoma cells. Materials and Methods: Cell viability was assessed by MTT assay and cell apoptosis through DAPI, TUNEL staining and Western blotting. Monodansylcadaverin(MDC) and AO staining were used to detect cell autophagy. Cell proliferation was assessed by Ki67 immunofluorescence staining. Results: Parthenolide induced growth inhibition in HepG2 cells. DAPI and TUNEL staining showed that parthenolide could increase the number of apoptotic nuclei, while reducing the expression of the anti-apoptotic protein Bcl-2 and elevating the expression of related proteins, like p53, Bax, cleaved caspase9 and cleaved caspase3. Parthenolide could induce autophagy in HepG2 cells and inhibited the expression of proliferation-related gene, Ki-67. Conclusions: Parthenolide can exert anti-cancer effects by inducing cell apoptosis, activating autophagy and inhibiting cell proliferation.

• The Journal of Korean Medicine
• /
• v.22 no.1
• /
• pp.90-95
• /
• 2001

목적：인진 약침액이 SK-N-MC 신경아세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향을 조사하였다. 방법 : SK-N-MC cell line에서의 아폽토시스 변화를 관찰하기 위해서 MTT assay, DAPI staining 및 flow cytometric analysis 방법을 이용하였다. 결과： MTT assay를 이용하여 분석한 결과 농도에 따른 세포 독성의 효과가 에탄올 투여로부터 관찰되었다. 또한 인진 약침액으로 전처치하고 에탄올을 처치하였을 때 세포 독성이 크게 감소되었다. DAPI staining에서 인진 약침액 투여군은 에탄올 투여군에 비해서 fragmentation이 억제되었다. Flow cytometry를 통하여 인진 약침액 투여군은 에탄올 투여군에 비하여 세포주기 중 sub $G_1$ 분획의 증가가 억제되었다. 결론 : SK-N-MC 신경아세포주에서 에탄올에 의해서 유발된 아폽토시스는 전형적인 세포사별 형태를 나타내었다. 또한 인진 약침액은 에탄올에 의해서 유발된 아폽토시스에서 세포보호 효과가 있음이 확인되었다.

• Journal of Acupuncture Research
• /
• v.29 no.1
• /
• pp.47-59
• /
• 2012

목적 : 이 연구는 봉독이 사람의 난소암 세포인 SKOV3와 PA-1에서 death receptor의 발현을 높여 세포자멸사를 촉진함으로써 암세포의 성장을 억제하는지 밝히고자 하였다. 방법 : 난소암의 세포자멸사의 관찰에는 DAPI, TUNEL staining assay를 시행하였으며, 세포자멸사 조절 단백질의 변동 관찰에는 western blot analysis를 시행하였고, 난소암 세포에서 death receptor의 변화를 관찰하기 위해 RT-PCR analysis를 시행하였다. 결과 : 1. DAPI, TUNEL staining assay 결과, 봉독은 투여량에 따라 세포자멸사의 유도를 통해 SKOV3와 PA-1 난소암세포의 증식을 억제하였고, 세포자멸사와 동반하여 DR4와 DR6의 발현이 두 암세포 모두에서 증가하였고, DR3의 출현은 PA-1 세포에서 증가하였다. 2. Death Receptor의 발현 증가에 따라 caspase-3, 8, 9 and Bax를 포함하는 세포자멸사 촉진 단백질의 발현이 동반하여 상승하였고 JAK2, STAT3의 인산화와 Bcl-2의 발현은 억제되었다. 3. siRNA 처리 시 봉독에 의한 DR3, DR4, DR6 발현증가와 STAT3의 활성억제가 역전되었다. 결론 : 이러한 결과는 봉독이 난소암 세포에서 DR3, DR4, DR6의 증가와 JAK2/STAT3 pathway의 억제를 통하여 세포자멸사를 유발한다는 것을 시사하며, 난소암의 예방과 치료에 효과적으로 활용될 수 있을 것으로 기대된다.

• Biomolecules & Therapeutics
• /
• v.20 no.5
• /
• pp.487-491
• /
• 2012

The aim of this study was to investigate the inhibitory effects of rutaecarpine on DNA strand breaks and apoptosis induced by hydrogen peroxide ($H_2O_2$) in murine Hepa-1c1c7 cells. Oxidative DNA damage was estimated by nuclear condensation assessment, fluorescence-activated cell sorting analysis, and Comet assay. Rutaecarpine inhibited cell death induced by $500{\mu}M$ $H_2O_2$, as assessed by 4',6-diamidino-2-phenylindole (DAPI) staining. Treatment with rutaecarpine reduced the number of DNA strand breaks induced by $H_2O_2$, as assessed by DAPI staining and Comet assay, and increased quinone reductase, phosphatidylinositol 3-kinase, and pAkt protein levels, as assessed by western blotting.