• 생태와환경
• /
• 제45권3호
• /
• pp.271-277
• /
• 2012

본 연구에서는 D. magna의 배양배지로서의 국내 자연수의 적절성을 평가하기 위하여, Elendt M4 배지에서 생산된 태어난 지 24시간 미만 된 어린 D. magna를 Elendt M4 배지, 탈염소 수도수 및 먹는 샘물에 21일 동안 노출시켜 생존율 및 번식능력을 평가하였다. 대조배지인 Elendt M4 배지와 먹는 샘물에서 배양한 D. magna는 어미의 생존율, 첫 배를 생산하는 시기, 생존한 어미 당 생산된 총 어린 물벼룩 평균수, 생존한 어미 당 생산된 죽은 어린 물벼룩 평균수는 2회의 번식시험 모두에서 Jonczyk과 Gilron (2005) 및 OECD No. 211, Daphnia magna Reproduction Test 지침서(OECD, 2008)의 기준을 벗어나지 않았다. 그러나 탈염소 수도수에서 배양을 한 경우에는 2번의 번식시험 모두 어미의 사망률이 20% 이상으로, 배양 13일, 15일, 18일에 사망된 개체가 관찰되었다. D. magna는 경도가 80 mg $CaCO_3\;L^{-1}$ 이상인 물에서 사용을 추천하고 있으나, 본 연구에서 사용된 탈염소 수도수의 경도는 50~53 mg $CaCO_3\;L^{-1}$ 이었다. 탈염소 수도수에서 나타난 지연된 사망률은 배양배지의 급격한 경도 차이에 의한 영향으로 판단된다. 그러므로 국내의 자연수(지하수, 표면수, 탈염소 수도수 등)를 사용하여 D. magna를 배양할 경우, 배양배지의 경도를 100 mg $CaCO_3\;L^{-1}$ 이상 강화시켜 사용하는 것이 필요하다. 그리고 궁극적으로는 국내에 서식하는 토착 물벼룩류를 대상으로 국내 수 환경에 적합한 시험생물을 개발하는 국가적인 연구가 필요하다고 사료된다.

• Toxicological Research
• /
• 제21권2호
• /
• pp.167-173
• /
• 2005

Bisphenol A (BPA) is a monomer used in the manufacture of a multitude of chemical products, including epoxy resins and polycarbonate. The objective of this study was to evaluate the effects of BPA administration on reproductive characteristics and blood hematological and chemical values in offspring of pregnant dams treated with BPA. BPA was administrated to pregnant mice by intraperitoneally injection with 0, 0.05, 0.5 and 5.0 mg/kg B.W. for 5 times at 3 days interval on gestation days 1-16. There were no treatment-related effects of BPA on reproductive organ weight in male offsprings at 45 days-of-age, but body weight was the lowest in 5.0mg BPA group when compared to other groups (P<0.05). No differences in semen characteristics (sperm concentration, viability, motility and abnormality) were observed between the control and BPA treatment groups. The WBC, HB, HT, MCV, MCH, MCHC, albumin, BUN and total protein of blood hematological and chemical values in male offsprings were not difference for any treatment groups, but RBC value in BPA groups was significantly increased comparing to the control group (P<0.05). The PLT value was slightly higher in 5.0mg BPA groups than in any other group, but not significantly difference among the experimental groups. In female offsprings, the effects of BPA didn't affect to the body and ovary weight, but the uterus weight in 5.0mg BPA group was slightly heavier than that of control group (P>0.05). No statically significant difference in blood hematological values in female offsprings were observed between the control group and BPA groups, but the concentration of albumin and BUN were significantly higher in 0.5mg BPA group when compared to control and other BPA treatment groups (P<0.05). The histological evaluation of testis and ovary in growing offspring at 45 days-of-age was not difference between the control group and BPA groups, but endometriosis of the uterus in female offspring was dramatically increased in 0.5 and 5.0mg BPA groups. These founding suggest that low concentration of BPA might not have a important role on reproductive ability or blood metabolite in offspring of pregnant dams treated with BPA.

• Toxicological Research
• /
• 제23권1호
• /
• pp.55-63
• /
• 2007

Diethylnitrosamine (DEN) is a nitrosamine compound that can induce a variety of liver lesions including hepatic carcinoma, forming DNA-carcinogen adducts. In the present study, microarray analyses were performed with Affymetrix Murine Genome 430A Array in order to identify the gene-expression profiles for DEN and to provide valuable information for the evaluation of potential hepatotoxicity. C57BL/6NCrj mice were orally administered once with DEN at doses of 0, 3, 7 and 20 mg/kg. Liver from each animal was removed 2, 4, 8 and 24 hrs after the administration. The histopathological analysis and serum biochemical analysis showed no significant difference in DEN-treated groups compared to control group. Conversely, the principal component analysis (PCA) profiles demonstrated that a specific normal gene expression profile in control groups differed clearly from the expression profiles of DEN-treated groups. Within groups, a little variance was found between individuals. Student's t-test on the results obtained from triplicate hybridizations was performed to identify those genes with statistically significant changes in the expression. Statistical analysis revealed that 11 genes were significantly downregulated and 28 genes were upregulated in all three animals after 2 h treatment at 20 mg/kg. The upregulated group included genes encoding Gdf15, JunD1, and Mdm2, while the genes including Sox6, Shmt2, and SIc6a6 were largely down regulated. Hierarchical clustering of gene expression also allowed the identification of functionally related clusters that encode proteins related to metabolism, and MAPK signaling pathway. Taken together, this study suggests that match with a toxicant signature can assign a putative mechanism of action to the test compound if is established a database containing response patterns to various toxic compounds.

• 해양환경안전학회지
• /
• 제26권5호
• /
• pp.514-522
• /
• 2020

해양생태독성시험의 국제표준시험종 중 생산자에 속하는 대표적인 Skeletonema sp.와 Dunaliella tertiolecta의 생태독성학적 차이점을 알아보기 위해 각 표준시험법(규격)을 비교하였고 환경에 대한 종 적합성과 다양한 시험물질에 대한 민감도를 비교 분석하였다. 그 결과, 시험법의 경우 대부분 동일하였으나 시험 유효성의 기준에서 pH 변화 제한과 초기접종밀도에서 차이가 나타났으며 이는 D. tertiolecta의 낮은 성장률에 기인된 것으로 추정된다. 적합성에서는 두 종 모두 규격에서 요구하는 유효성의 기준을 연속 만족하여 시험수행의 일관성을 보였고 시험한계 염분범위는 Skeletonema sp.와 D. tertiolecta 각각 20 및 10 psu로 나타났다. 마지막으로 민감도의 경우, 시험규격에서 제시하는 참조물질, 실제 오염 배출수(선박평형수) 및 기타 다양한 화학물질에서 모두 Skeletonema sp.가 D. tertiolecta에 비해 독성 민감도가 상대적으로 높음을 확인하였으며 이는 해양생태독성시험 수행에 있어 생산자를 이용한 시험의 경우 최소 2종 이상의 다른 분류군의 미세조류를 이용하는 것이 시험결과의 신뢰성과 객관성을 높일 수 있는 방법임을 시사한다.

• Toxicological Research
• /
• 제20권1호
• /
• pp.71-82
• /
• 2004

Changes in cell cycle control in the lungs and liver of the B6C3F1 mice (20 males per each group) exposed to ozone (0.5 ppm), 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK, 1.0 mg/kg), and dibutyl phthalate (DBP, 5,000 ppm) after 52 weeks were examined through Western, Northern blot, and immunohistochemistry based on alterations in protein expression levels of G1/S checkpoints (cyclin D1, cyclin E, and PCNA), G2/M checkpoints (cyclin B1, cyclin G, and cyclin A), negative regulators (p53, p21, GADD45, and p27), and positive regulator (mdm2). Expression levels of cyclins D1, E, G, PCNA, mutant p53, and mdm2 proteins were higher in the lungs and livers treated with combination of toxicants than in those treated with ozone only. Expression levels of the wild-type and mutant p53, p21, GADD45, p27, and mdm2 proteins and mRNAs were higher in toxicant-treated groups than those of the control. Immunohistochemical analysis revealed staining intensities of the PCNA, cyclin D1, c-myc and mdm2 protein- treated lungs and livers were stronger than those of the control group. Our results showed that combined treatment of ozone with NNK/DBP altered the cell cycle control through instability of the wild-type p53 gene. Such pivotal p53-mediated cell cycle alterations may be responsible for the toxicity observed under our experimental condition. These results may be applied to risk assessment of mixture-induced toxicity.

• Toxicological Research
• /
• 제13권4호
• /
• pp.339-347
• /
• 1997

Disulfiram (DSF) and diethyldithiocarbamate (DDC), a reduced form of DSF, protect the liver against toxicant-induced injury through inhibition of cytochrome P450 2E1. The effect of DSF and DDC on the levels of major hepatic microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) expression was comparatively studied, given the view that these enzymes are involved in terminal detoxification events for high energy intermediates of xenobiotics. Treatment of rats with a single dose of DSF (20-200 mg/kg, po) resulted in 2- to 15-fold increases in the mEH mRNA level at 24 hr with the ED$_{50}$ value being noted as 60 mg/kg. The mEH mRNA level was elevated ~15-fold at 24 hr after treatment at the dose of 100 mg/kg, whereas the hepatic mRNA level was rather decreased from the maximum at the dose of 200 mg/kg, indicating that DSF might cause cytotoxicity at the dose. In contrast to the effect of DSF, DDC only minimally elevated the mEH mRNA level at the doses employed. DSF moderately increased the major GST mRNA levels in the liver as a function of dose, resulting in rGSTA2, rGSTA3/5 or rGSTM1 mRNA levels being elevated 3- to 4-fold at 24 hr post-treatment, whereas the rGSTM2 mRNA level was not altered. DDC, however, failed to stimulate the mRNA levels for major GST subunits, indicating that the reduced form of DSF was ineffective in stimulating the GST the expression. The effect of other organosulfides including aldrithiol, 2, 2'-dithiobis(benzothiazole) (DTB), tetramethylthiouram disulfide (TMTD) and allyl disulfide (ADS) on the hepatic mEH and GST mRNA expression was assessed in rats in order to further confirm the increase in the gene expression by other disulfides. Treatment of rats with aldrithiol (100 mg/kg, po) resulted in a 16-fold increase in the mEH mRNA level at 24 hr post-treatment. DTB, TMTD and ADS also caused 5-, 9- and 12-fold increases in the rnRNA level, respectively, as compared to control. Thus, all of the disulfides examined were active in stimulating the mEH gene in the liver. The organosulfides significantly increased the rGSTA2, rGSTA3, rGSTA5 and rGSTM1 mRNA levels at 24 hr after administration. In particular, aldrithiol was very efficient in stimulating the rGSTA and rGSTM genes among the disulfides examined. These results provide evidence that DSF and other sulfides effectively stimulate the mEH and major GST gene expression at early times in the liver and that DDC, a reduced form of DSF, was ineffective in stimulating the expression of the genes, supporting the conclusion that reduced form(s) of organosulfur compound(s) might be less effective in inducing the mEH and GST genes through the antioxidant responsive element(s).

• Toxicological Research
• /
• 제6권2호
• /
• pp.205-213
• /
• 1990

The regulation of neurotoxicants has usually been based upon setting reference doses by dividing a no observed adverse effect level (NOAEL) by uncertainty factors that theoretically account for interspecies and intraspecies extraploation of experimental results in animals to humans. Recently, we have proposed a four-step alternative procedure which provides quantitative estimates of risk as a function of dose. The first step is to establish a mathematical relationship between a biological effect or biomarker and the dose of chemical administered. The second step is to determine the distribution (variability) of individual measurements of biological effects or their biomarkers about the dose response curve. The third step is to define an adverse or abnormal level of a biological effect or biomarker in an untreated population. The fourth and final step is to combine the information from the first three steps to estimate the risk (proportion of individuals exceeding on adverse or abnormal level of a biological effect or biomarker) as a function of dose. The primary purpose of this report is to enhance the certainty of the first step of this procedure by improving our understanding of the relationship between a biomarker and dose of administered chemical. Several factors which need to be considered include: 1) the pharmacokinetics of the parent chemical, 2) the target tissue concentrations of the parent chemical or its bioactivated proximate toxicant, 3) the uptake kinetics of the parent chemical or metabolite into the target cell(s) and/or membrane interactions, and 4) the interaction of the chemical or metabolite with presumed receptor site(s). Because these theoretical factors each contain a saturable step due to definitive amounts of required enzyme, reuptake or receptor site(s), a nonlinear, saturable dose-response curve would be predicted. In order to exemplify this process, effects of the neurotoxicant, methlenedioxymethamphetamine (MDMA), were reviewed and analyzed. Our results and those of others indicate that: 1) peak concentrations of MDMA and metabolites are ochieved in rat brain by 30 min and are negligible by 24 hr, 2) a metabolite of MDMA is probably responsible for its neurotoxic effects, and 3) pretreatment with monoamine uptake blockers prevents MDMA neurotoxicity. When data generated from rats administerde MDMA were plotted as bilolgical effect (decreases in hippocampal serotonin concentrations) versus dose, a saturation curve best described the observed relationship. These results support the hypothesis that at least one saturable step is involved in MDMA neurotoxicity. We conclude that the mathematical relationship between biological effect and dose of MDMA, the first step of our quantitative neurotoxicity risk assessment procedure, should reflect this biological model information generated from the whole of the dose-response curve.

• Biomolecules & Therapeutics
• /
• 제22권6호
• /
• pp.510-518
• /
• 2014

Chronic (>24 h) exposure of arsenite, an environmental toxicant, has shown the decreased nitric oxide (NO) production in endothelial cells (EC) by decreasing endothelial NO synthase (eNOS) expression and/or its phosphorylation at serine 1179 ($eNOS-Ser^{1179}$ in bovine sequence), which is associated with increased risk of vascular diseases. Here, we investigated the acute (<24 h) effect of arsenite on NO production using bovine aortic EC (BAEC). Arsenite acutely increased the phosphorylation of $eNOS-Thr^{497}$, but not of $eNOS-Ser^{116}$ or $eNOS-Ser^{1179}$, which was accompanied by decreased NO production. The level of eNOS expression was unaltered under this condition. Treatment with arsenite also induced reactive oxygen species (ROS) production, and pretreatment with a ROS scavenger N-acetyl-L-cysteine (NAC) completely reversed the observed effect of arsenite on $eNOS-Thr^{497}$ phosphorylation. Although protein kinase C (PKC) and protein phosphatase 1 (PP1) were reported to be involved in $eNOS-Thr^{497}$ phosphorylation, treatment with PKC inhibitor, Ro318425, and overexpression of various PKC isoforms did not affect the arsenite-stimulated $eNOS-Thr^{497}$ phosphorylation. In contrast, treatment with PP1 inhibitor, calyculin A, mimicked the observed effect of arsenite on $eNOS-Thr^{497}$ phosphorylation. Lastly, we found decreased cellular PP1 activity in arsenite-treated cells, which was reversed by NAC. Overall, our study demonstrates firstly that arsenite acutely decreases NO production at least in part by increasing $eNOS-Thr^{497}$ phosphorylation via ROS-PP1 signaling pathway, which provide the molecular mechanism underlying arsenite-induced increase in vascular disease.

• 생태와환경
• /
• 제55권3호
• /
• pp.185-200
• /
• 2022

수환경 내 다양한 유해물질의 위해성에 대한 관심은 환경 매체 내 물질의 농도뿐만 아니라 복잡한 먹이단계를 통한 어류 체내의 축적과 어류를 통한 인체 위해성으로 이어진다. 국내의 경우 2016년 이후 생활에서 사용되고 있는 화학제품(생활화학제품) 기인 위해 우려물질의 관리를 위한 등록과 평가 등에 관한 법률 개정과 함께 이들 물질의 환경 배출이 주목받게 됨에 따라 수생태계 내 잔류여부에 대한 조사도 수행되기 시작했다. 최근에는 이러한 물질의 관리를 위한 생태계 내 분포 조사 및 배출 계수 산정을 위한 연구사업이 수행되고 있는데 해당 연구 사업에서는 세정제, 접착제, 염색제, 방향제 등을 비롯한 화장품이나 세제 등에 포함되는 성분과 살균·소독제를 대상으로 영양단계 내 축적과 전달을 이해하기 위한 물질의 축적과 확대를 포함한다. 본 논문은 최근 발표된 생활화학제품기인 유해물질의 수환경 유입 및 분포에 대한 연구 결과를 정리하고 그 과학적 의미를 제시하며 또한 국내외 수행되고 있는 수환경 모니터링 기법에 대한 연구의 예를 바탕으로 현재 유해화학물질의 수환경 내 잔류 농도 및 분포, 생태계 모니터링을 위한 연구의 방향을 제안하고자 한다. 특히 어류를 대상으로 하는 조사에서 국내 수역에 서식하는 주요 어류조사 및 이를 바탕으로 한 대상 어류 선정의 필요성과 인체 위해성 연구의 필요성 등 시기적으로 요구되는 연구를 위한 영양단계 해석과 생물확대계수 연구의 방향을 소개하며 향후 국내에서 수행되고 있는 생물상 모니터링과 화학물질 연구에 대한 제언을 포함한다.