• Parasites, Hosts and Diseases
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• 제57권5호
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• pp.489-497
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• 2019

Cystic echinococcosis (CE), a zoonotic disease caused by Echinococcus granulosus at the larval stage, predominantly develops in the liver and lungs of intermediate hosts and eventually results in organ malfunction or even death. The interaction between E. granulosus and human body is incompletely understood. Exosomes are nanosized particles ubiquitously present in human body fluids. Exosomes carry biomolecules that facilitate communication between cells. To the best of our knowledge, the role of exosomes in patients with CE is not reported. Here, we isolated exosomes from the sera of patients with CE (CE-exo) and healthy donors and subjected them to liquid chromatography-tandem mass spectrometry analysis. Proteomic analysis identified 49 proteins specifically expressed in CE-exo, including 4 proteins of parasitic origin. The most valuable parasitic proteins included tubulin alpha-1C chain and histone H4. And 8 proteins were differentially regulated in CE-exo (fold change>1.5), as analyzed with bioinformatic methods such as annotation and functional enrichment analyses. These findings may improve our understanding about the interaction between E. granulosus and human body, and may contribute to the diagnosis and prevention of CE.

• Asian Pacific Journal of Cancer Prevention
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• 제16권5호
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• pp.1833-1837
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• 2015

An aristolactam-type alkaloid, isolated from Orophea enterocarpa, is enterocarpam-III (10-amino-2,3,4,6-tetramethoxyphenanthrene-1-carboxylic acid lactam). It is cytotoxic to various human and murine cancer cell lines; however, the molecular mechanisms remain unclear. The aims of this study were to investigate cytotoxic effects on and mechanism (s) of human cancer cell death in human hepatocellular carcinoma HepG2 and human invasive breast cancer MDA-MB-231 cells compared to normal murine fibroblast NIH3T3 cells. Cell viability was determined by MTT assay to determine $IC_{10}$, $IC_{20}$ and $IC_{50}$ levels, reactive oxygen species (ROS) production with 2',7'-dichlorohydrofluorescein diacetate and the caspase-3, -8 and -9 activities using specific chromogenic (p-nitroaniline) tetrapeptide substrates, viz., DEVD-NA, IETD-NA and LEHD-NA and employing a microplate reader. Mitochondrial transmembrane potential (MTP) was measured by staining with 3, 3'-dihexyloxacarbocyanine iodide ($DiOC_6$) and using flow cytometry. The compound was cytotoxic to HepG2 and MDA-MB-231 cells with the $IC_{50}$ levels of $26.0{\pm}4.45$ and $51.3{\pm}2.05{\mu}M$, respectively. For murine normal fibroblast NIH3T3 cells, the $IC_{50}$ concentration was $81.3{\pm}10.1{\mu}M$. ROS production was reduced in a dose-response manner in HepG2 cells. The caspase-9 and -3 activities increased in a concentration-dependent manner, whereas caspase-8 activity did not alter, indicating the intrinsic pathway activation. Enterocarpam-III decreased the mitochondrial transmembrane potential (MTP) dose-dependently in HepG2 cells, suggesting that the compound induced HepG2 cell apoptosis via the mitochondrial pathway. In conclusion, enterocarpam-III inhibited HepG2 and MDA-MB-231 cell proliferation and induced human HepG2 cells to undergo apoptosis via the intrinsic (mitochondrial) pathway and induction of caspase-9 activity.

• Journal of Life Science
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• 제11권1호
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• pp.57-64
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• 2001

The cDNA encoding human NeuAc ${\alpha}$2,3Gal$\beta$ 1,3GalNAc GalNac ${\alpha}$2,6-Sialyltransferase (hST6GalNac IV) was isolated by screening of human fetal liver cDNA library with a DNA probe generated from the cDNA sequence of mouse ST6Gal NAc IV (mkST6GalNAc IV). The cDNA sequence included an open reading frame coding for 302 amino acids, and comparative analysis of this cDNA with mST6GalNAc IV showed that each sequence of the predicted coding region contains 88% and 85% identifies in nucleotide and amino acid levels, respecively. The primary structure of this enzyme suggested a putative domain structure, like that in other glycosyltransferases, consisting of a short N-terminal cytoplamic domain, a transmembrane domain and a large C-terminal active domain. This enzyme expressed in COS-7 cells echibited transferase activity toward NeuAc ${\alpha}$2,3Gal$\beta$ 1,3GalNAc, fetuin and GM1b, although the activity toward the later is very low, no significant activity being detected toward Gal${\beta}$ 1,3Gal NAc or asialofetuin, the other glycoprotein substrates tested. The $^{14}$ C-sialylated residue of fetuin sialylated by this enzyem with CMP-[$^{14}$C]NeuAc was sensitive to treatment with ${\alpha}$2,8-specific sialidase of Vibrio cholerae but resistant to treatment with ${\alpha}$2,3-specific sialidase (NaNase I), and ${\alpha}$2,3- and ${\alpha}$2,8-specific sialidase of Newcastle disease virus. These results clearly indicated that the expressed enzyme is a type of GalNAc ${\alpha}$2,6-sialyltransferase like mST6GalNAc IV, which requires sialic acid residues linked to Gal${\beta}$1,3GalNAc-residues for its activity.

• 한국지하수토양환경학회지:지하수토양환경
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• 제9권1호
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• pp.12-27
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• 2004

본 연구에서는 벤젠을 대상으로 오염된 지하수를 생활용수로 사용했을 때 발생하는 실내오염도를 모사하고 실내에서 가능한 흡입, 섭취, 피부흡수와 같은 노출경로를 고려하여 노출시나리오를 자성하였다. 인체에 유입된 벤젠에 대하여 PBPK 모델을 이용하여 인체의 각 장기에 어떻게 분포하는지를 분석하였다. 결과에서 흡입과 섭취가 주요노출 경로였으며 남성이 여성보다 많은 호흡량으로 인해 보다 높은 노출속도를 유지하였다. 노출속도에 대한 피부흡수의 공헌도는 상대적으로 매우 작았다. 단기노출의 결과 오염물 노출에 대하여 SPT, RPT,간의 정맥혈 중 벤젠농도는 빠르게 증감하는 반면 지방의 경우는 느리게 반응하였고 많은 벤젠이 지방세포에 축적되어 정맥혈에는 적은 농도로 존재하였다. 장기간 노출에서 여성은 남성보다 전체적으로 2.1배 많은 벤젠을 체내에 축적하고 있는 것으로 나타났다. 장기간 노출에서 총유입벤젠의 98%가 호흡과 대사분해에 의해 제거되었다. 흡입경로는 벤젠이 호흡배출에 의해 69.8% 제거되었으며 섭취경로는 48.4%로 오염물이 유입되는 위치에 따라 각각의 제거기작의 공헌도가 다르게 나타났다. 본 연구의 결과는 실내오염에 따라 오염물이 체내에 흡수되고 분포ㆍ제거되는 현상을 이해하고 노출저감대책을 마련하는데 필요한 자료를 제공하고자 하였다.

• 한국축산식품학회지
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• 제23권2호
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• pp.137-144
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• 2003

본 연구에서는 IGFs를 다양한 기능성 유제품에 적용하기 전에 먼저 상업용 IGFs 및 초유로부터 분리.정제된 IGFs의 안전성을 평가하여 새로운 기능성 소재로서의 IGFs의 가능성을 검토해 보고자 하였다. 이를 위하여 독성평가에서 기본적으로 수행되고 있는 단회투여독성시험과 반복투여독성시험을 실시하였다. 단회투여독성시험에서는 R&D Systems사의 Recombinant Human IGF- I 을 시험물질로 하고 생후 4주된 Sprague-Dawley계(♂) 흰쥐(rat)를 실험동물로 하여 개체당 0, 10, 20, 50 $\mu\textrm{g}$씩 투여하는 시험군을 설정한 후, 꼬리정 꼬리정맥에 주사하여 20일간 관찰하면서 체중변화, 식이섭취변화를 측정하고 최종적으로 부검하여 육안검사 및 간장, 신장, 비장에 대한 병리조직검사를 실시한 결과, 명확한 이상소견이 나타나지 않아 모든 시험군에서 급성독성이 확인되지 않았다. 반복투여독성시험에서는 초유로부터 분리.정제된 IGF- I 을 시험물질로 하고 생후 4주된 Sprague-Dawley계(♂) 흰쥐(rat)를 실험동물로 하여 개체당 0, 5, 10, 15 $\mu\textrm{g}$씩 투여하는 시험군을 설정한 후, 14일간 반복적으로 강제경구투여하면서 임상증상을 관찰하면서 체중변화, 식이섭취변화를 측정하였고 최종적으로 부검하여 육안검사 및 간장, 신장, 비장에 대한 병리조직검사를 실시한 결과, 명확한 이상소견이 나타나지 않아 모든 투여농도에서 반복투여독성시험을 비롯한 기본적인 독성평가에 있어서 초유 내 IGF-I의 안전성에는 명확한 이상소견이 없는 것으로 판단되었다.

• Applied Microscopy
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• 제29권1호
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• pp.43-56
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• 1999

태생기 간 적혈구형성 중 혈관밖공간의 미성숙 적혈구모세포의 일부는 동굴모세혈관 속공간으로 이주하며, 별큰포식세포에 의해 영향을 받는다. 본 연구에서는 혈관속공간에 이주한 미성숙 적혈구모세포와 별큰포식세포의 관계를 규명하고자 간 적혈구형성의 활성도가 높게 유지되는 태령 제 11주부터 태령 제 20주에 이르는 태아 간과 쥐 태자 간의 연구재료를 대상으로 투과 및 주사전자현미경 관찰을 통해 다음과 같은 결론을 얻었다. 1) 별큰포식세포는 성인 간이나 다른 태령 시기에 비해 비대하였고 많은 세포질돌기들을 갖고 있었다. 이 세포는 부분적으로 세포분열에 의해 증식되고 있었으며 성인 간의 별큰포식세포와 다르게 자가종식을 할 수 있었다. 3) 호산성적혈구모세포에서 탈출된 핵은 별큰포식세포의 속공간쪽과 간세포쪽 세포막에서 포식되었다. 포식된 핵은 주변 부위에 층판이 형성되거나 및 개의 작은 조각으로 파괴되는 등 다양한 소화과정을 보였다. 3) 동굴모세혈관 속공간에서 별큰포식세포는 미성숙적혈구모세포들과 직접으로 접촉하거나, 미성숙 적혈구 모세포들 사이에 별큰포식세포의 긴 세포질돌기가 뻗쳐 있어 골수 적혈구형성에서 관찰되는 적혈구모세포섬과 비슷하였다. 이상의 결과를 종합하면 태아 간 적혈구형성이 왕성한 시기의 별큰포식세포는 적혈구를 포식하는 것 외에도, 세포가 비대해져서 동굴모세혈관을 통한 혈액흐름을 감소시키고, 적혈구모세포섬을 구성하여 태아 간 적혈구형성에서 미성숙 적혈구모세포의 성숙과 관련된 기계적 및 생리학적 기능을 수행한다고 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.6991-6996
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• 2015

In the past decade, the incidence and mortality rates of cholangiocarcinoma (CCA) have been increasing worldwide. The relatively low responsiveness of CCA to conventional chemotherapy leads to poor overall survival. Recently, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or Apo2L) has emerged as the most promising anti-cancer therapeutic agent since it is able to selectively induce apoptosis of tumor cells but not normal cells. In this study, we aimed to investigate the therapeutic effect of TRAIL in CCA cell lines (M213, M214 and KKU100) compared with the immortal biliary cell line, MMNK1, either alone or in combination with a subtoxic dose of 5-fluorouracil (5-FU). We found that recombinant human TRAIL (rhTRAIL) was a potential agent which significantly inhibited cell proliferation and mediated caspase activities (caspases 8, 9 and 3/7) and apoptosis of CCA cells. The combined treatment of rhTRAIL and 5-FU effectively enhanced inhibition of CCA cell growth with a smaller effect on MMNK1. Our finding suggests TRAIL to be a novel anti-cancer therapeutic agent and advantage of its combination with a conventional chemotherapeutic drug for effective treatment of CCA.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.214-218
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• 2003

Alcohol consumption causes numerous consequences on the health of the human body. Heavy drinking on a daily base has caused liver diseases. Furthermore, some products such as acetaldehyde produced from alcohol metabolism are more toxic than alcohol itself. This study was carried out to evaluate the role of Lactobacillus casei on alcohol metabolism, especially, the removal of the toxic effect of alcohol. The maximum alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities from L. casei were observed at 4 hr of culture. L. casei was confirmed to produce the ADH and ALDH by the SDS-PAGE. From in vivo test using SD rats with 22% alcoholic drink, blood alcohol concentration (BAC), glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) of the rats feeding the medium containing L. casei were lower than those of the rats feeding the medium containing an alcoholic drink only This demonstrates that the ADH and ALDH produced by L. casei have virtual functions to detoxicate the alcohol in vivo and the fermentation broth of L. casei can be used as an alcohol detoxification drink.

• BMB Reports
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• 제29권2호
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• pp.146-150
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• 1996

A 2.1 kb cDNA clone for rat transketolase was isolated from rat liver ${\lambda}gt11$ cDNA library and its sequence was determined. The predicted rat transketolase (655 amino acids with $M_r$ 71,186) is highly similar (92%) to that of the human enzyme except that it contains an extra 32 amino acids at its N-terminus. Although it is less similar (<27%) to transketolases from non-mammalian species, the functional motifs such as the catalytic sites and thiamine binding domain are well conserved in the rat enzyme. Southern blot analysis of genomic DNA verified that transketolase appears to be derived from a single gene. Immunoblot and Northern blot analyses suggested that hepatic transketolase was activated pretranslationally by a 2.1-fold while little change was observed in brain enzyme, indicating a tissue-specific pretranslational activation during postnatal development.

• Toxicological Research
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• 제17권
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• pp.79-82
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• 2001

The infection of liver flukes, Clonorchis sinensis (CS) and Opisthorchis viverrini (OV), has been known as a risk factor to induce cholangiocellular carcinoma (CCC) in human living in the endemic area, providing promoting effect on the liver initiated by chemical carcinogens. The present study evaluated the relationship between the dosage level of dimethylnitrosamine (DMN) and the infection load of CS in the neoplastic development by histopathological examination of the treated hamsters. To evaluate the effects of DMN, different doses of DMN ranging from 0 to 25 ppm were administered to hamsters with 20 CS metacercariea. For the risk assessment of the infection load, 0, 5, 15, 50 CS metacercariae were respectively infected with 12 ppm DMN. The mortality was closely related to the infection load rather than the concentration of DMN. The infection of CS clearly promoted the induction of CCC even at dose level of 6 ppm DMN. Only five metacercariae were enough to promote CCC induction at the concentration of 12 ppm DMN.