• Food Science and Biotechnology
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• 제17권6호
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• pp.1322-1326
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• 2008

Tannase catalyzes the hydrolysis of gallic acid esters and hydrolysable tannins. Twenty-two Lactobacillus strains with tannase activity were isolated from 7 types of kimchi. A polymerase chain reaction-based assay targeting the recA gene assigned all isolates to either Lactobacillus plantarum or Lactobacillus pentosus. The tannase activities of isolates measured in whole cells and cell-free extracts varied even within each species. The activities of the isolates varied with the assay method, but both methods indicated that isolate LT7 (identified as L. pentosus) showed the highest activity. The results of thin layer chromatography and high performance liquid chromatography, respectively, showed that tannic acid and gallic acid degraded to pyrogallol in resting L. pentosus LT7 cells. Therefore, the putative biochemical pathway for the degradation of tannic acid by L. pentosus implies that tannic acid is hydrolyzed to gallic acid and glucose, with the formed gallic acid being decarboxylated to pyrogallol. This study revealed the possible production of pyrogallol from tannic acid by the resting cell reaction with L. pentosus LT7.

• 한국자원식물학회지
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• 제31권4호
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• pp.342-354
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• 2018

Suaeda japonica is a halophytic plant that lives in mudflat at intertidal zone of western and southern coastal areas of Korea. The seawater-living plants showed a purple color during their whole life. In contrast, freshwater-living plants displayed a green color in leaves. When seawater-living plants were transferred to potting soil, the purple color was gradually changed to green in the leaves. The extracted purple pigment compound exhibited typical characteristics of betacyanin that were represented by water solubility, pH- and temperature-dependent color changes, sensitivity to light, UV-Vis spectra, and gel electrophoretic migration pattern. The LC-MS analysis of the extracted pigment compound showed the presence of two major protonated molecular ions ($[M+H]^+$) at m/z 651.1 and m/z 827.1. Antioxidant activity of the pigment compound was determined using stable free radical DPPH assay. It was found to have an antioxidant activity that is linearly increased in proportion to the reaction time for up to 30 min, and the activity was comparable to that of control BHA at 9.0 mg/ml. The anticancer activity against several tumor cell lines was also examined following the MTT assay. The significant growth inhibitory effect was observed on two tumor cell lines, SW-156 (human kidney carcinoma) and HEC-1B (human endometrial adenocarcinoma). Probably, the pigment compound may function as an osmolyte to uphold halotolerant physiological processes in saline environment.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.94-94
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• 2003

Endocrine disrupters (EDs) are exogenous chemicals that interfere with the production, releasing, metabolism, excretion, binding of natural hormones, and whole endocrine systems. EDs are very dangerous since they are extremely stable, not easily degraded, and accumulated in fat and tissue. 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) is known as the most toxic EDs. Therefore, this study was conducted to investigate the effects of TCDD on proliferation of human breast cancer (MCF-7) and endometrial adenocarcinoma (Hec-1B) cells. 10, 100, and 1000 nM of TCDD were treated with steroid free condition. Viable cell counting, MTT, and BrdU assay was performed to investigate cell proliferation. Apoptosis was investigated using DNA laddering. Although, DNA fragmentation as the evidence of apoptosis was not detected, all of these cell lines showed restricted proliferation at 48 hrs after 100 and 1000 nM TCDD treatments. Recently, it has been reported that the expression of transforming growth factor $\beta$s (TGF-$\beta$s) are increased in TCDD treatment and also involved in regulation of cell cycle. Therefore, these results were considered that the decreased cell prolifcration by TCDD is related to the expression of TGF-$\beta$s.

• Toxicological Research
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• 제12권1호
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• pp.69-79
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• 1996

Primary culture of cerebellar neuronal cells derived from 8-day old Long-Evans rats was used. Pure granule cells, astrocytes or mixed cells culture systems were prepared. These cells were differentiated and developed synaptic connections. And the astrocytes were identified by immunostaining with glial fibrillary acidic protein (GFAP). Methamphetamine (MAP), which acts on dopaminergic system and cadmium (Cd), a toxic heavy metal, were applied and biochemical assays and electrophysiological studies were performed. $LC_50$ values estimated by MTT assay of MAP and Cd were 3 mM and 2$\mu M$ respectively. Cells were treated with 1 mM or 2 mM MAP and 1$\mu M$ $CdCl_2$ for 48 hour, and the incubation media were analyzed for the content of released LDH. MAP (2 mM) and Cd significantly increased the LDH release. Cell viability was decreased in both groups and some cytopathological changes like cell swelling or vacuolization were seen. The cerebellar granule cells were used for measuring membrane currents using whole-cell clamp technique. Sodium and potassium currents were not affected by MAP neither Cd, but calcium current was significantly reduced by Cd but not affected by MAP. Therefore, in vitro neurotoxicity test system using neuronaI cells and astrocytes cultures were established and can be used in screening of potential neurotoxic chemicals.

• The Korean Journal of Physiology and Pharmacology
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• 제23권5호
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• pp.393-402
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• 2019

Aurora kinases inhibitors, including ZM447439 (ZM), which suppress cell division, have attracted a great deal of attention as potential novel anti-cancer drugs. Several recent studies have confirmed the anti-cancer effects of ZM in various cancer cell lines. However, there have been no studies regarding the cardiac safety of this agent. We performed several cytotoxicity, invasion and migration assays to examine the anti-cancer effects of ZM. To evaluate the potential effects of ZM on cardiac repolarisation, whole-cell patch-clamp experiments were performed with human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and cells with heterogeneous cardiac ion channel expression. We also conducted a contractility assay with rat ventricular myocytes to determine the effects of ZM on myocardial contraction and/or relaxation. In tests to determine in vitro efficacy, ZM inhibited the proliferation of A549, H1299 (lung cancer), MCF-7 (breast cancer) and HepG2 (hepatoma) cell lines with $IC_{50}$ in the submicromolar range, and attenuated the invasive and metastatic capacity of A549 cells. In cardiac toxicity testing, ZM did not significantly affect $I_{Na}$, $I_{Ks}$ or $I_{K1}$, but decreased $I_{hERG}$ in a dose-dependent manner ($IC_{50}$: $6.53{\mu}M$). In action potential (AP) assay using hiPSC-CMs, ZM did not induce any changes in AP parameters up to $3{\mu}M$, but it at $10{\mu}M$ induced prolongation of AP duration. In summary, ZM showed potent broad-spectrum anti-tumor activity, but relatively low levels of cardiac side effects compared to the effective doses to tumor. Therefore, ZM has a potential to be a candidate as an anti-cancer with low cardiac toxicity.

• 동의생리병리학회지
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• 제20권5호
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• pp.1285-1289
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• 2006

Hominis placenta (HP) has been used as an agent for promoting physiological function in traditional asian medicine. The present study was peformed to investigate whether HP acupuncture treatment in an experimental tumor mice model inhibit tumor growth through immunomodulatory effects. Mice were inoculated subcutaneously with colon26-L5 cells on the back. Three days after tumor inoculation, HP herbal acupuncture treatment was conducted on BL18 acupoint every other day for three weeks. HP Herbal acupuncture treatment significantly suppressed the primary tumor growth and prolonged survival rate. To evaluate immunomodulatory effect of HP acupuncture, splenocytes proliferation assay, fluorescence-activated cell sorting (FACS) and ELISA for IFN- ${\gamma}$, and IL-4 cytokine level. HP herbal acupuncture enhanced the mitogenic activity of Balb/c whole splenocytes induced by various mitogenic stimuli and increased immune cell population such as T cell, B cell, Th cell, Tc cell and Macrophages. HP herbal acupuncture caused a marked increase of production of Th1 cytokine (IFN- ${\gamma}$ ,) and decrease of production of Th2 cytokine (IL-4). These results indicated that HP herbal acupuncture suppresses tumor growth through a mechanism leading to a Th1 dominant immune state.

• Applied Biological Chemistry
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• 제49권3호
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• pp.227-232
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• 2006

미더덕(Styela clava)은 척색동물문 미색동물아문에 속하는 해양생물로서, 독특한 향과 맛으로 인해 식품에 널리 이용되고 있다. 본 연구에서는 미더덕의 생리활성물질로의 활용가능성을 탐색하고자 미더덕을 전체, 살 및 껍질 부분으로 나누어 아세톤으로 추출한 뒤 산화적 DNA 손상억제 및 항암 활성을 측정하였다. 미더덕 각 부위 추출물을 5, 10, $50\;{\mu}g/ml$의 농도로 백혈구에 처리한 후 200\;{\mu}M의 $H_2O_2$로 산화적 스트레스를 유발하여 DNA 손상 억제 정도를 검증하기 위해 comet assay를 실시하였다. 미더덕 전체 추출물을 5, 10, $50\;{\mu}g/ml$의 농도로 백혈구에 처리했을 때 손상된 DNA tail 부분의 DNA 함량을 측정한 % fluorescence in tail이 26.9, 27.0, 23.8%로 63.9%인 $H_2O_2$ 처리 양성대조구에 비해 유의적으로 감소하였으며 미더덕살 추출물의 경우, 5, 10, $50\;{\mu}g/ml$ 처리시 농도에 의한 효과 차이는 볼 수 없었지만 양성대조구에 비해 유의적으로 DNA 손상정도가 감소하였다. 미더덕 껍질 추출물의 경우 같은 농도로 처리했을 때 각각 농도 의존적이며 유의적으로 DNA 손상정도가 감소하였다. 인간 대장암 유래의 세포주 HT-29의 성장억제 효과에 조사하기 위해 미더덕 부위별 아세톤 추출물을 10, 50, 100, $500\;{\mu}g/ml$의 농도로 각각 처리한 뒤 MTT reduction assay 방법으로 측정한 결과, 각 추출물에 대해 전체적으로 농도의존적으로 암세포 성장 억제효과가 증가되었다. 전체, 살, 껍질 부위별 추출물을 $100\;{\mu}g/ml$의 농도로 첨가하였을 때 각각 90.5%, 82.0%, 75.2%로 비교적 낮은 활성을 보였지만, $500\;{\mu}g/ml$로 처리했을 때 각각 26.9%, 30.6%, 12.0%로 급격히 활성이 증가하는 것을 볼 수 있었다. 특히 껍질 부분의 아세톤 추출물이 강한 DNA 손상억제 및 암세포 성장 억제 효과를 보임을 알 수 있었다.

• 한국어병학회지
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• 제10권2호
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• pp.75-86
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• 1997

어류의 감염 조직으로부터 edwardsiellosis의 원인균인 Edwardsiella tarda를 whole cell 자체로 직접 검출할 수 있는 solid phase ELISA법을 연구하였다. A. hydrophila ATCC7966, V. anguillarum HUFP5001, Y. ruckeri 11-4, E. ictaluri 및 Streptococcus sp. NG8206 등의 어병세균에 대해 ELISA법으로 실시한 교차반응 분석에서 A. hydrophila ATCC7966 균주와 V. anguillarum HUFP5001 균주가 E. tarda Edk-2에 대한 토끼 항혈청에 대해 높은 교차반응을 나타내었으나, 항혈청을 A. hydrophila ATCC7966 FKC로 흡착시킴으로써 교차반응을 제거할 수가 있었다. 그러나, 응집항체가 측정 결과와는 달리, ELISA 분석에서는 E. tarda 분리 균주간의 교차반응이 매우 높은 것으로 나타났다. Tissue homogenate내에 있는 항원을 검출함에 있어, 조직내의 지질이나 단백질 성분이 함께 분석용 plate에 coating되어 감도가 훨씬 감소하므로 ELISA법의 적용을 위해서는 감염 조직의 homogenate를 PBS에 100배 이상 희석한 후 진단을 실시해야 하는 것으로 나타났다. Tissue homogenate내에 있는 생균을 항원으로 하여 직접 검출할 때에는 검출한계가 $1{\times}10^3$ cells/ml로 나타나 FKC 항원의 사용에 비하여 더 증가된 감도를 보여주었다. 결론적으로 본 ELISA법은 양식장에서 발생한 edwardsiellosis를 진단함에 있어서 특이적이고 신속하며 민감한 방법으로 확인되었다.

• Radiation Oncology Journal
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• 제3권1호
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• pp.9-12
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• 1985

To determine the radiosensitivity and dose-survival characteristics of jejunal crypt cells, experimental study was done using total 40 mice. Single irradiation of 1,000 rad to 1,600rad was delivered to whole bodies of mice, using a cesium 137 animal irradiator. The number of regenerating crypts per jejunal circumference was counted, by using a jejunal crypt cell assay technique, and dose response curve was measured. The average number of jejunal crypt Per circumference in control group was $140\pm10$. Mean lethal dose$(D_0)$ of moose jejunal crypt cell was 135rad.

• Journal of Periodontal and Implant Science
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• 제23권1호
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• pp.1-15
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• 1993

P. gingivalis has been implicated as a strong pathogen in periodontal disease and known to have three serotypes of P. gingivalis. The purpose of this study is to investigate on the relationship between virulence, metabolic acids and genetic heterogeneity of P. gingivalis. P. gingivalis W50 standard strain and five strains of P. gingivalis serotype b Korean isolates were used in this study. For in vitro virulence test, lyophilized whole cell P. gingivalis were suspended, and sonicated with ultrasonic dismembranometer. Sonicated samples were applied to cultured cells derived from periodontal ligament, and cell activity was assayed with growth and survival assay. The metabolic acids were also extracted, and determined by High Performance Liquid Chromatography. Pst I-digested bacterial genomic DNA was electrophoresed, and densitometric analysis was performed to study the genetic heterogeneity. All of the P. gingivalis serotype b produced butyric acid. In cell activity study, butyric acid inhibited the cell activity irrespective of its concentration. Densitometric analysis showed restriction fragment length polymorphism. These results suggested that there existed heterogeneity of the metabolic acids and the virulence of P. gingivalis and such heterogeneity might be related to genetic heterogeneity.