• Ocean and Polar Research
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• v.31 no.4
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• pp.349-357
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• 2009

Ballast water is widely recognized as a serious environmental problem due to the risk of introducing non-indigenous aquatic species. In this study we aimed to investigate measures which can minimize the transfer of aquatic organisms from ballast water. Securing more reliable technologies to determine the viability of aquatic organisms is an important initiative in ballast water management systems. To evaluate the viability of marine phytoplankton, we designed the staining methods of fluorescein diacetate (FDA) and Calcein-AM assay on each target species belonging to different groups, such as bacillariphyceae, dinophyceae, raphidophyceae, chrysophyceae, haptophyceae and chlorophyceae. The FDA method, which is based on measurements of cell esterase activity using a fluorimetric stain, was the best dye for determining live cells of almost all phytoplankton species, except several diatoms tested in this study. On the other hand, although fluorescence of Calcein-AM was very clear for a comparatively longer time, green fluorescence per cell volume was lacking in most of the tested species. According to the Flow CAM method, which is a continuous imaging technique designed to characterize particles, green fluorescence values of stained cells by FDA were significantly higher than those of Calcein-AM treatments and control, implying that the Flow CAM using FDA assay could be adapted as an important tool for distinguishing living cells from dead cells. Our results suggest that the FDA and Calcein-AM methods can be adapted for use on phytoplankton, though species-specific characters are greatly different from one organism to another.

• International Journal of Oral Biology
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• v.43 no.2
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• pp.61-68
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• 2018

Codium fragile (Suringar) Hariot is an edible green seaweed that belong to the Codiaceae family and has been used in Oriental medicine for the treatment of enterobiasis, dropsy, and dysuria. Methanol extract of codium fragile has anti-oxidant, anti-inflammatory and anti-cancer properties, although the anti-cancer effect on oral cancer has not yet been reported. In this study, we investigated the anti-cancer activity and the mechanism of cell death by methanol extracts of Codium fragile (MeCF) on human FaDu hypopharyngeal squamous carcinoma cells. Our data showed that MeCF inhibits cell viability in a dose-dependent manner, and markedly induced apoptosis, as determined by the MTT assay, Live/Dead assay, and DAPI stain. In addition, MeCF induced the proteolytic cleavage of procaspase -3, -7, -9 and poly(ADP-ribose) polymerase(PARP), and upregulated or downregulated the expression of mitochondrial-apoptosis factor, Bax(pro-apoptotic factor), and Bcl-2(anti-apoptotic factor). Futhermore, MeCF induced a cell cycle arrest at the G1/S phase through suppressing the expression of the cell cycle cascade proteins, p21, CDK4, CyclinD1, and phospho-Rb. Taken together, these results indicated that MeCF inhibits cell growth, and this inhibition is mediated by caspase- and mitochondrial-dependent apoptotic pathways through cell cycle arrest at the G1/S phase in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, methanol extracts of Codium fragile can be provided as a novel chemotherapeutic drug due to its growth inhibition effects and induction of apoptosis in human oral cancer cells.

• International Journal of Oral Biology
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• v.45 no.3
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• pp.107-114
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• 2020

Acacetin, which is present in damiana (Turnera diffusa) and black locust (Robinia pseudoacacia), has several pharmacologic activities such as antioxidant, anti-inflammatory, and anti-proliferative effects on cancer cells. However, the effect of acacetin on head and neck cancers has not been clearly established. This study aimed to examine the effects of acacetin on cell growth and apoptosis induction in FaDu human pharyngeal carcinoma cells. These were investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, Live/Dead cell assay, 4',6-diamidino-2-phenylindole dihydrochloride staining, caspase-3 and caspase-7 activation assay, and immunoblotting in FaDu cells. Acacetin induced FaDu cell death in a dose-dependent manner, with an estimated IC₅₀ value of 41.9 µM, without affecting the viability of L-929 mouse fibroblasts as normal cells. Acacetin treatment resulted in nuclear condensation in the FaDu cells. It promoted the proteolytic cleavage of procaspase-3, -7, -8, and -9 with increasing amounts of the cleaved caspase isoforms in FaDu cells. Acacetin-induced apoptosis in FaDu cells was mediated by the expression of Fas and activation of caspase-8, caspase-3, and poly (ADP-ribose) polymerase. Immunoblotting showed downregulation of the anti-apoptotic mitochondrial proteins Bcl-2 and Bcl-xL, but upregulation of the mitochondria-dependent pro-apoptotic proteins Bax and Badin FaDu cells after acacetin treatment. These findings indicate that acacetin inhibits cell proliferation and induces apoptotic cell death in FaDu human pharyngeal carcinoma cells via both the death receptor-mediated extrinsic apoptotic pathway and the mitochondria-mediated intrinsic apoptotic pathway.

• Korean Journal of Environmental Biology
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• v.27 no.1
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• pp.66-72
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• 2009

Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.

• Natural Product Sciences
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• v.13 no.3
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• pp.234-240
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• 2007

The present study investigated the anti-tumor activity of ethyl acetate extract of Acanthospermum hispidum DC against daltons ascites lymphoma in mice. The extract was prepared by cold maceration with ethyl acetate for 3 - 7 days and evaporated in vacuum to dry. (Yield : 14.2 g, 1.42% w/w). The extract was fractionated by column chromatography by using gradient elution technique and the diterpenes fraction isolated (0.649 g). Both extract and the fraction were administered as oral suspension with tween 20 in water to tumor bearing mice (DAL) and changes in dead cell count, histopathology of tumor cells, hematological parameters and median survival time (MST) were examined and compared with that of tumor control or 5-Fluorouracil (5-FU). The results indicate that both ethyl acetate extract and fraction possess anti-tumor activity. The study suggests that Acanthospermum hispidum DC seems promising as a source of diterpenes for potential anti-tumor activity.

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.6
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• pp.1562-1565
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• 2008

HCMCO5 is a herbal extract which comprises of eight different herbs. We studied whether this prescription has an acute toxicity in rats. SPF Sprague-Dawley male and female rats were administered orally with HMC05 extract of 2,000 mg/kg for 14days. We examined mortality, clinical signs, body weights and gross findings during the tests. The result showed no dead animals. We also could not find a significant body weight changes during the experimental period. In addition, no differences were found between control and HMC05 treated groups in clinical signs and other findings. These results indicate that HMC05 extract did not show any toxic effects, and oral ADL value was over 2,000 mg/kg in SD rats.

• International Journal of Oral Biology
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• v.30 no.2
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• pp.65-69
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• 2005

Periodontitis is a multi-microbial disease and the comparison of a series of periodontopathogenic and non-periodontopathogenic bacteria in terms of microbe-host interaction may provide clues to understand the microbial etiology of the disease better. When we deal with twenty different bacterial species in a study, the first technical issue is how to measure the accurate concentration and use the same number of bacterial cells. We measured bacterial concentration by enumerating bacteria stained with SYTOX green for constant time using a flow cytometer and compared the results with those obtained by plate counting. Concentrations calculated by two different methods were very close. Therefore, flow cytometric counting allowed the rapid analysis of live/dead bacteria, offering the advantage of turbidity measurement and that of colony counting together.

• Korean Journal of Pharmacognosy
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• v.35 no.2 s.137
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• pp.122-127
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• 2004

This study was conducted to investigate the safety and efficacy of an oriental herbal composition, Kamihonghwatang(KH-19), for the reduction of the side effects of chemotherapeutic drug. KH-19 prevented the reduction of white blood cells including lymphocytes, monocytes and eosinophiles in C57BL/6 mice injected with fluorouracil, a commonly used anticancer drug. KH-19 also prevented the reduction of cell densities in bone marrow and spleen of fluorouracil-injected mice. To evaluate the safety of KH-19, single-dose toxicity test was conducted using SD rats. No dead animal was found and the minimum lethal dose of KH-19 was more than 5000 mg/kg.

• Journal of Korean Neurosurgical Society
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• v.38 no.5
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• pp.387-389
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• 2005

Primary atypical teratoid/rhabdoid tumor[AT/RT] of the central nervous system is a recently described, highly malignant neoplasm that usually occur in the posterior fossa of children. Although AT/RT usually occurs in young children, AT/RT is being recognized in adults with increasing frequency. The authors report 49-year-old man with multiple AT/RT lesions [right lateral ventricle, right temporal lobe and right cerebellum]. Histopathologic findings showed typical rhabdoid cells with eccentric nuclei and prominent nucleoli. Eventhough the tumor was removed, a patient was dead in one month after surgery due to recurrence and rapid regrowth of the tumor.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.2
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• pp.167-172
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• 2013

In order to establish the diagnostic indices of skin diseases, some physiological and pathological concepts of interstices(腠理) were researched based on , and western dermatology with etymological analysis. As physiological indices for diagnosis of skin diseases, measurement of epidermis and dermis using ultrasonogram in the zhongwan and dachui's location. And for grading looseness and fineness of interstices with 3 unsound groups, measuring numbers and sizes of sweat pores in each point's 1 cm diameter circular area using comparative method and palpation assessment. Another index is superficial temperature. As pathological indices for diagnosis, validating volumes of dead skin cells and grading degrees of atrophy and degeneration of skin lesion. And as supplementary measures, absorptive degrees of cosmetics on face should be recorded according to 3 grades. These diagnostic indices can contribute to establishment of standard pattern identification and prescription of skin diseases through converting anatomical cognizance into classical concepts of interstices objectively.