• Advances in Traditional Medicine
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• 제8권3호
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• pp.228-235
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• 2008

This study was conducted to evaluate the estrogen activity of silkworm (Bombyx mori) pupa extracts and their fractions. Powdered samples of freeze-dried silkworm pupa were extracted at room temperature (RT), $40^{\circ}C$, $60^{\circ}C$, $80^{\circ}C$, and $100^{\circ}C$ in water (D.W), chloroform, ethyl acetate, and methanol for 6h and then filtered (0.45 um). The extracts were then freeze-dried. The estrogenic activity of these extracts was then investigated by competition binding assays using estrogen receptor ${\alpha}\;(ER{\alpha})$ and $ER{\beta}$, and by evaluating their effects on the proliferation of the human breast cancer cell line, MCF-7. Among the extracts evaluated, water extracts prepared at RT showed the highest binding affinity to $ER{\alpha}$ ($IC_{50}$, 1.76 ug/ml) and $ER{\beta}$ ($IC_{50}$, 0.07 ug/ml). In addition, MCF-7 cells that were treated with 62.5 ug/ml of the RT extract showed the greatest increase in proliferation (2-fold; 1291.79%) when compared to control cells (659.82%). Next, the water extract that was prepared at RT (sample 1) was dissolved in D.W. and further fractionated using a Dowex 50W - 8X ($H^+$) column. The flow-through and wash were then pooled together and freeze-dried (sample 2). The bound materials were then eluted with 20 mM NaCl, after which they were applied to a Dowex 1X2 - 200 ($Cl^-$) column and washed with D.W. to remove the sodium ions. The eluants were then freeze-dried (sample 3). Of these fractions, sample 2 showed the highest binding affinity to ER{\alpha} ($IC_{50}$, 1.44 ug/ml) and $ER{\beta}$ ($IC_{50}$, 1.18 ug/ml). In addition, MCF-7 cells that were treated with sample 2 (15.6 ug/ml) showed the largest increase in growth (1159.39%) when compared to control cells (525.26%). Taken together, these results suggest that the fraction of the RT water extract of silkworm pupa referred to as sample 2 may be useful as a phytoestrogen.

• Toxicological Research
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• 제25권4호
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• pp.175-180
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• 2009

Lead (Pb) is a ubiquitously occurring environmental heavy metal which is widely used in industry and human life. Possibly due to a global industrial expansion, recent studies have revealed the prevalent human exposure to Pb and increased risk of Pb toxicity. Once ingested by human, 95% of absorbed Pb is accumulated into erythrocytes and erythrocytes are known to be a prime target for Pb toxicity. Most of the studies were however, focused on $Pb^{2+}$ whereas the effects of $Pb^{4+}$, another major form of Pb on erythrocytes are poorly understood yet. In this study, we investigated and compared the effects of $Pb^{4+}$, $Pb^{2+}$ and other heavy metals on procoagulant activation of erythrocytes, an important factor for the participation of erythrocytes in thrombotic events in an effort to address the cardiovascular toxicity of $Pb^{4+}$. Freshly isolated erythrocytes from human were incubated with $Pb^{4+}$, $Pb^{2+}$, $Cd^{2+}$ and $Ag^+$ and the exposure of phosphatidylserine (PS), key marker for procoagulant activation was measured using flow cytometry. As a result, while $Cd^{2+}$ and $Ag^+$ did not affect PS exposure, $Pb^{4+}$ and $Pb^{2+}$ induced significantly PS exposure in a dose-dependent manner. Of a particular note, $Pb^{4+}$ induced PS exposure with a similar potency with $Pb^{2+}$. PS bearing microvesicle (MV), another important contributor to procoagulant activation was also generated by $Pb^{4+}$. These PS exposure and MV generation by $Pb^{4+}$ were well in line with the shape change of erythrocyte from normal discocytes to MV shedding echinocytes following $Pb^{4+}$ treatment. Meanwhile, nonspecific hemolysis was not observed suggesting the specificity of $Pb^{4+}$-induced PS exposure and MV generation. These results indicated that $Pb^{4+}$ could induce procoagulant activation of erythrocytes through PS exposure and MV generation, suggesting that $Pb^{4+}$ exposure might ultimately lead to increased thrombotic events.

• Nutrition Research and Practice
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• 제9권2호
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• pp.129-136
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• 2015

BACKGROUND/OBJECTIVES: A variety of immunomodulators can improve the efficacy of low-dose chemotherapeutics. Active hexose correlated compound (AHCC), a mushroom mycelia extract, has been shown to be a strong immunomodulator. Whether AHCC could enhance the antitumor effect of low-dose 5-fluorouracil (5-FU) via regulation of host immunity is unknown. MATERIALS/METHODS: In the current study Hepatoma 22 (H22) tumor-bearing mice were treated with PBS, 5-FU ($10mg{\cdot}kg^{-1}{\cdot}d^{-1}$, i.p), or AHCC ($360mg{\cdot}kg^{-1}{\cdot}d^{-1}$, i.g) plus 5-FU, respectively, for 5 d. $CD^{3+}$, $CD^{4+}$, $CD^{8+}$, and NK in peripheral blood were detected by flow cytometry. ALT, AST, BUN, and Cr levels were measured by biochemical assay. IL-2 and $TNF{\alpha}$ in serum were measured using the RIA kit and apoptosis of tumor was detected by TUNEL staining. Bax, Bcl-2, and TS protein levels were measured by immunohistochemical staining and mRNA level was evaluated by RT-PCR. RESULTS: Diet consumption and body weight showed that AHCC had no apparent toxicity. AHCC could reverse liver injury and myelosuppression induced by 5-FU (P < 0.05). Compared to mice treated with 5-FU, mice treated with AHCC plus 5-FU had higher thymus index, percentages of $CD^{3+}$, $CD^{4+}$, and NK cells (P < 0.01), and ratio of $CD^{4+}$/$CD^{8+}$ (P < 0.01) in peripheral blood. Radioimmunoassay showed that mice treated with AHCC plus 5-FU had the highest serum levels of IL-2 and $TNF{\alpha}$ compared with the vehicle group and 5-FU group. More importantly, the combination of AHCC and 5-FU produced a more potent antitumor effect (P < 0.05) and caused more severe apoptosis in tumor tissue (P < 0.05) compared with the 5-FU group. In addition, the combination of AHCC and 5-FU further up-regulated the expression of Bcl-2 associated X protein (Bax) (P < 0.01), while it down-regulated the expression of B cell lymphoma 2 (Bcl-2) (P < 0.01). CONCLUSIONS: These results support the claim that AHCC might be beneficial for cancer patients receiving chemotherapy.

• Nutrition Research and Practice
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• 제1권2호
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• pp.105-112
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• 2007

Ascorbic acid has been reported to extend replicative life span of human embryonic fibroblast (HEF). Since the detailed molecular mechanism of this phenomenon has not been investigated, we attempted to elucidate. Continuous treatment of HEF cells with ascorbic acid at ($200{\mu}M$) from 40 population doubling (PD) increased maximum PD numbers by 18% and lowered $SA-{\beta}-gal$ positive staining, an aging marker, by 2.3 folds, indicating that ascorbic acid extends replicative life span of HEF cells. Ascorbic acid treatment lowered DCFH by about 7 folds and Rho123 by about 70%, suggesting that ascorbic acid dramatically decreased ROS formation. Ascorbic acid also increased aconitase activity, a marker of mitochondrial aging, by 41%, indicating that ascorbic acid treatment restores age-related decline of mitochondrial function. Cell cycle analysis by flow cytometry revealed that ascorbic acid treatment decreased G1 population up to 12%. Further western blot analysis showed that ascorbic acid treatment decreased levels of p53, phospho-p53 at ser 15, and p21, indicating that ascorbic acid relieved senescence-related G1 arrest. Analysis of AP (apurinic/apyrimidinic) sites showed that ascorbic acid treatment decreased AP site formation by 35%. We also tested the effect of hydrogen peroxide treatment, as an additional oxidative stress. Continuous treatment of $20{\mu}M$ of hydrogen peroxide from PD 40 of HEF cells resulted in premature senescence due to increased ROS level, and increased AP sites. Taken together, the results suggest that ascorbic acid extends replicative life span of HEF cells by reducing mitochondrial and DNA damages through lowering cellular ROS.

• Journal of Microbiology and Biotechnology
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• 제20권2호
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• pp.438-445
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• 2010

The immunomodulatory effects of Aureohasidium pullulans SM-2001 exopolymers containing $\beta$-1,3/1,6-glucan were evaluated in cyclophosphamide (CPA)-treated mice. To induce immunosuppression, 150 and 110 mg/kg of CPA were intraperitoneally injected 3 days and 1 day, respectively, before beginning administration of the test material. Exopolymers were delivered subcutaneously or orally, four times, in a volume of 10 ml/kg at 12-h intervals beginning 24 h after the second CPA treatment. Changes in thymus and spleen weights, splenic amounts of tumor necrosis factor (TNF)-$\alpha$, interleukin (IL)-$1{\beta}$, and IL-10, and numbers of CD3+, CD4+, CD8+, and TNF-$\alpha+$ thymus and spleen cells were monitored in CPA-treated mice. As a result of CPA treatment, dramatic decreases in the number of CD3+, CD4+, CD8+, and TNF-$\alpha+$ cells were detected in the thymus and spleen, along with decreases in thymus and spleen weights. In addition, splenic TNF-$\alpha$, IL-$1{\beta}$, and IL-10 contents were also decreased on observation with flow cytometry. However, oral and subcutaneous treatments with exopolymers effectively reduced the immunosuppressive changes induced by CPA. Therefore, it is concluded that exopolymers of A. pullulans SM-2001 can effectively prevent immunosuppression through, at least partially, the recruitment of T cells and TNF-$\alpha+$ cells or enhancement of their activity, and can provide an effective component of prevention or treatment regimens for immunosuppression related to cancer, sepsis, and high-dose chemotherapy or radiotherapy.

• KSBB Journal
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• 제27권5호
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• pp.308-312
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• 2012

Argon-plasma jet (Ar-PJ) is generated by ionizing Ar gas, and the resulting Ar-PJ consists of a mixture of neutral particles, positive ions, negative electrons, and various reactive species. Although Ar-PJ has been used in various biomedical applications, little is known about the biological effects on cells located near the plasma-exposed region. Here, we investigated the effects of the Ar-PJ on actin cytoskeleton of mouse embryonic fibroblasts (MEFs) in response to indirect as well as direct exposure to Ar-PJ. This Ar-PJ was generated at 500 mL/min of flow rate and 100 V electric power by our device mainly consisting of electrodes, dielectrics, and a high-voltage power supply. Because actin cytoskeleton is the key cellular machinery involved in cellular movement and is implicated in regulation of cancer metastasis and thus resulting in a highly desirable cancer therapeutic target, we examined the actin filament architectures in Ar-PJ-treated MEFs by staining with an actin-specific phalloidin labeled with fluorescent dye. Interestingly, the Ar-PJ treatment causes destabilization of actin filament architectures in the regions indirectly exposed to Ar-PJ, but no differences in MEFs treated with Ar gas alone and in untreated cell control, indicating that this phenomenon is a specific cellular response against Ar-PJ in the live cells, which are indirectly exposed to Ar-PJ. Collectively, our study raises the possibility that Ar-PJ may have potential as anti-cancer drug effect through direct destabilization of the actin cytoskeleton.

• Journal of Preventive Medicine and Public Health
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• 제33권1호
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• pp.117-124
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• 2000

목적 : 이 연구의 목적은 적출간 관류법을 통하여 염화니켈과 염화카드뮴의 간 독성을 비교하는 데에 두었다. 방법 : AST(aspartate aminotransferase ferase), ALT(alanine aminotransferase), LDH(lactate dehydrogenase) 등의 간기능을 나타내는 생화학적 지표와 관류속도를 간독성 지표로 이용하였으며 산소 소모율은 생존력을 나타내는 지표로 이용하였다. $300({\pm}50)g$ 정도의 흰쥐를 대조군과 각 노출군($50{\mu}M$ 및 $200{\mu}M$의 염화니켈과 염화카드뮴)에 5마리씩 총 25마리를 무작위로 할당하였다. Krebs-Ringer bicarbonate buffer solution을 관류액 용도로 제조하여 간문맥에 투입하고 간세포를 통과하여 vena cava로 배출되는 관류액을 일정한 시간 간격을 두고 계속적으로 채취하였으며 채취한 관류액은 간독성 지표를 측정하는 데에 이용하였다. 결과 : AST, ALT, LDH 모두 염하니켈 보다는 염화카드뮴 투여군에서 시간의 경과에 따라 큰 폭으로 증가하였으며 반복 측정된 2요인 분산분석을 실시한 결과, 통계적으로 유의한 차이를 나타냈다. 생존력은 두 물질 모두에서 시간의 경과에 따라 감소하는 경향을 나타냈다. 결론 : 결론적으로, 이 실험상에서는 염화니켈 보다 염화카드뮴의 간독성이 더욱 강한 것으로 드러났다. 적출간 더욱 강한 것으로 드러났다. 적출간 관류법의 특성상 급성 간독성 평가에서 적절한 기법으로 보이며 국내 연구에서도 이 방법이 널리 이용되기를 기대한다.

• 인터넷정보학회논문지
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• 제17권2호
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• pp.19-28
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• 2016

최근 정보통신망의 발전과 스마트 폰의 대량 보급으로 인하여 인터넷 트래픽이 기하급수적으로 증가하고 있다. 이와 관련하여, 본 논문은 증가하고 있는 인터넷 침해사고와 네트워크 공격 중 대표적인 DDoS 공격에 대해서 탐지 및 분석한다. 이를 위해 네트워크 플로우 정보를 바탕으로 동작할 수 있도록 기존의 ATMSim 분석 패키지의 기능과 GUI를 개선하고, 이를 이용하여 캠퍼스 내부 LAN을 통해 대량으로 유입되는 정상적인 트래픽과 DDoS 공격이 포함된 비정상 트래픽을 생성한다. 수집 생성된 정상 비정상 트래픽의 특성을 분석하기 위해서 자기유사성 추정 기법을 이용하여, 그래픽 분석 및 Hurst 파라메터 (자기유사성 파라메터) 추정량 분석결과 정상 트래픽과 비정상 트래픽이 자기유사성 관점에서 추정치 Hurst 값이 높음을 보여 주고 있다.

• 원예과학기술지
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• 제32권3호
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• pp.366-374
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• 2014

이배체 탱자로부터 자연적으로 발생한 동질 사배체 탱자의 수체 형질 관련 표현형 및 유전체 메틸화 변이 정도를 분석하여 후성 유전의 하나인 유전체 메틸화가 동질 사배체의 표현형 변이의 요인으로 작용할 수 있음을 구명하고자 본 실험을 수행하였다. 이배체 탱자에서 유래된 14주의 사배체 탱자로부터 염색체를 분석하여 이수성이 없는 2n = 4X = 36 식물체로 확인하였다. CMA 핵형 분석 결과 염색체가 배가된 동질 사배체임을 확인할 수 있었다. 동질 사배체에서 수고, 수폭, 원가지 수, 원가지 길이, 분지 각도, 마디 길이, 잎의 특성 등 동질 사배체 수체 표현형에 있어서 상당한 변이가 나타남을 확인하였다. 또한 동질 사배체 광합성률에는 큰 차이는 없었지만, SPAD 값에 의한 엽록소 지수에 있어서도 표현형이 다양하게 나타나는 것을 알 수 있었다. 그 외에도 기공 밀도와 공변 세포 길이에 있어서 광범위하게 변이가 관찰되는 것을 알 수 있었다. Global cytosine DNA 메틸화를 분석한 결과 개체 간 메틸화 정도에 차이가 존재함을 알 수 있었다. 동질 사배체 탱자 14주의 절반이 이배체 탱자의 메틸화와 비교하였을 때 2배 이상으로 나타난 것을 확인하였다. 본 연구 결과 동질 사배체에서 나타나는 수체형질 변이가 gene redundency를 줄이기 위한 global cytosine DNA 메틸화와 관계될 수 있음을 확인하였다.

• Journal of Pharmaceutical Investigation
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• 제38권6호
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• pp.373-380
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• 2008

In order to develop optimal formulation and iontophoresis condition for the transdermal delivery of levodopa, we have evaluated the effect of two permeation enhancers, ethanol and oleic acid in microemulsion, on transdermal delivery of levodopa. In vitro flux studies were performed at $33^{\circ}C$, using side-by-side diffusion cell and full thickness hairless mouse skin. Current density applied was $0.4\;mA/cm^2$ and current was off after 6 hours application. Levodopa was analysed by HPLC at 280 nm. The o/w microemulsions of oleic acid in buffer solution (pH 2.5 & 4.5) were prepared using oleic acid, Tween 80 and ethanol. The existence of microemulsion regions were investigated in pseudo-ternary phase diagrams. Contrary to our expectation, cumulative amount of levodopa transported from microemulsion (pH 2.5) for 10 hours was similar to that from aqueous solution in all delivery methods (passive, anodal and cathodal). When pH of the micro-emulsion was pH 4.5, cumulative amount of levodopa transported for 10 hours increased about 40% (anodal) to 50% (cathodal), when compared to that from aqueous solution. Flux from pH 4.5 microemulsion showed higher value than that from pH 2.5 in all delivery methods. These results seem to indicate that electroosmosis plays more dominant role than electrorepulsion in the flux of levodopa at pH 2.5. The effect of ethanol on iontophoretic flux was studied using pH 2.5 phosphate buffer solution containing 3% or 5% (v/v) ethanol. Flux enhancement was observed in passive and anodal delivery as the concentration of the ethanol increased. Without ethanol, cathodal delivery showed higher flux than anodal delivery. Anodal delivery increased the cumulative amount of levodopa transported 1.6 fold by 5% ethanol after 10 hours. However, in cathodal delivery, no flux enhancement of levodopa was observed during current application and only marginal increase in cumulative amount transported after 10 hours was observed by 5% ethanol. These results seem to be related to the decrease in dielectric constant of the medium and the lipid extraction of the ethanol, which decrease the electroosmotic flow, and thus decrease the flux. Overall, the results provide important insights into the role of electroosmosis and electrorepulsion in the transport of levodopa through skin, and provide some useful informations for optimal formulation for levodopa.