• Environmental Analysis Health and Toxicology
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• v.15 no.3
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• pp.61-67
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• 2000

Toxic lesions of styrene in the Japanese Medaka (Oryzias latipes) were compared with those of styrene oxide, the active metabolite of styrene, using embryo-larval assays. The developmental stages of Japanese Medaka (Oryzias latipes) treated with both chemicals were not altered and progressed normally. However, styrene oxide was more toxic than styrene in terms of causing death and lesions . High concentrations of styrene (higher than 4.9 ppm) and styrene oxide (higher than 2.4 ppm), resulting in more than 50% mortality, caused similar lesions of cardiovascular system, craniofacial bone formation and spinal deformities, although a number of lesions were not observed by both chemicals . In the group treated with styrene, eyeball sizes and intereye distances were reduced, while, in the group treated with styrene oxide , the eyes and eye cups were not developed and two eyes were sometimes fused. In addition, styrene oxide caused the lesion which involved the posterior brain and brain stem were herniated through the spinal cord . The noticeable difference of toxic symptoms between these two chemicals was the time of onset. Toxicities of cardiovascular system and craniofacial bone formation appeared on day 3 of development in styrene oxide treated group, but, styrene treated group staned to show hemorrhages on day 3 and the craniofacial malformation were appeared on day 5, These differences between two chemicals may be due to the metabolism of styrene to styrene oxide, the reactive intermediate.

• Journal of Embryo Transfer
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• v.8 no.1
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• pp.1-7
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• 1993

This study was carried out to investigate the survival rates of late mouse molulae frozen in the state of vitrification and then thawed after equilibrating them separately in EFS 40, GFS 40 and DFS 40 at 1$0^{\circ}C$. The results obtained are as follows : 1. Freezing in the state of vitrification and thawing late mouse molulae after equilibrating them at l0$0^{\circ}C$ in EFS 40 for 30 seconds, one minute and two minutes, we obtained survival rates of 76.7%, 96.7% and 100%, respectively. 2. Freezing and thawing them after equilibrating at 1$0^{\circ}C$ in GFS 40 for 30 seconds, one minute and two minutes, we obtained survival rates of 60%, 96.7% and 10%, respectively. These results are as similar as in the case of EFS 40. 3. Freezing and thawing them after equilibrating at l$0^{\circ}C$ in DFS 40 for 30 seconds and one minute, we obtained survival rates of 62.1% and 0%, respectively. These results represent lower survival rates than those obtained with EFS 40 and GFS 40. In conclusion, even equilibrating late mouse molulae in EFS 40 and GFS 40 at 1$0^{\circ}C$ for more than one minute gives a survival rate of more than 97%, while equilibrating them in DFS 40 at 1$0^{\circ}C$ for more than one minute results in a 0% survival rate, which means that DFS 40 has a strong toxicity.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.283-289
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• 2000

Objective: The study was performed to compare the survival rate and the development of day 2 mouse embryos which had freezing procedures done. Methods: We used three different vitrification solutions (EFS, VS14, DPS) and a ultrarapid freezing solution (UFS) for cryopreservation of day 2 mouse embryo. Results: We tested toxicity by exposing embryos to vitirification solutions and a ultrarapid freezing solution. The survival rates are 100%, 97.8%, 95.6% and 100% (EFS, VS14, DPS and UFS). After cultured for 96 hours, hatching rates of each group are 93.5% (no freezing), 95.6% (EFS), 86.4% (VS14), 93.0% (DPS), and 93.0% (UFS). There is no significant differences among groups. The survival rates after thawing cryopreserved embryos are 80.2%, 91.7%, 69.5%, 0% and 91.8% (slow freezing, EFS, VS14, DPS and UFS). Also cultured for 96 hours, the hatching rates are 93.5% (no freezing), 84.1% (slow freezing), 93.9% (EFS), 48.5% (VS14) and 70.1% (UFS). Conclusion: The survival rates of vitrification in EFS solution and ultrarapid freezing are higher than slow freezing (p<0.05). The hatching rate of vitrification in EFS solution cultured for 96 hours is highest, so vitrification of day 2 mouse embryos in EFS solution considered as more effective for cryopreservation.

• Toxicological Research
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• v.29 no.1
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• pp.27-34
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• 2013

This study was conducted to investigate the potential embryo-fetal toxicity and toxicokinetics of the antimalarial agent artesunate (ARTS) in Sprague-Dawley rats. Pregnant rats were administered ARTS daily from gestational day 6~15 via oral gavage, at test doses of 0, 2, 4, or 8 mg/kg (22 females per group). The fetuses were examined for external, visceral, and skeletal abnormalities on gestational day 20. With regard to the dams, there were no deaths, treatment-related clinical signs, changes in body weight, or food intake in any of the treatment groups. There were no treatment-related gross findings at necropsy in any treatment group. In the 8 mg/kg group, there was a decrease in gravid uterine weight and in the weight of female fetuses. There was also an increase in fetal deaths (primarily late resorptions) and an increase in post-implantation losses (37%) at 8 mg/kg. An increase in the incidence of visceral and skeletal variations at 4 and 8 mg/kg was observed. These defects included minor changes in the appearance of the kidney and thymus, as well as absent ribs or thoracic vertebrae. Toxicokinetics were assessed in a parallel study, using 4 mated females per group. Using liquid chromatography-mass spectrometry (LC-MS) analysis, the concentration of ARTS and its metabolite dihydroartemisinin (DHA) were quantified in plasma from rats on gestational days 5, 6, 10, and 15. Amniotic fluid was assayed for ARTS and DHA on gestational day 15. There was evidence of rapid conversion of ARTS to the metabolite DHA in maternal plasma, since ARTS could not be consistently detected in plasma at the three doses tested. ARTS and DHA were not detected in amniotic fluid at gestational day 15, indicating limited placental transfer of the two agents. The embryo-fetal no-observable-adverse-effect level (NOAEL) of the test item was considered to be 8 mg/kg/day for dams, and 2 mg/kg/day for embryo-fetal development.

• Environmental Analysis Health and Toxicology
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• v.22 no.3
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• pp.211-218
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• 2007

Even though some standard developmental bioassay protocols for environmental assessment using sea urchins have already been described, there have not been many attempts to apply and modify these protocols with Korean species. Therefore, there is a strong need to establish standard bioassay protocols using sea urchins commonly found in Korea. Prior to developing a new protocol, it is essential to know the optimal conditions for the bioassay procedures. We investigated the optimal conditions (temperature, salinity, and embryo density) of the sea urchin Strongylocentrotus intermedius. The ideal temperature for developmental bioassay of S. intermedius was determined to be $15^{\circ}C$ and the time required for the embryo to become pluteus larva was 72 hr. The optimal range of salinity for the embryo toxicity using S. intermedius was between 30 to 32 psu, which is similar to the range found in the natural habitats of adult populations. The optimum density of embryos at the beginning of bioassays was 100 embryos/mL. When the assays were carried out at higher densities, the proportion of normally developed larvae decreased significantly.

• Korean Journal of Environmental Biology
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• v.30 no.3
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• pp.231-237
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• 2012

Toxicity of $Ni^{2+}$ and Tebuconazole were investigated via FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol using domestic frog embryos. Embryos of Black-spotted pond frog, Rana nigromaculata, were incubated and toxic effects of $Ni^{2+}$ and Tebuconazole were investigated by probit analysis. As a result, mortality and malformation rates were increased and larval body length was decreased in a dose dependant manner of $Ni^{2+}$ and Tebuconazole. The half maximal effective concentration ($EC_{50}$) of $Ni^{2+}$ and Tebuconazole were 0.07, $12.7mg\;L^{-1}$, respectively and the half maximal lethal concentration ($LC_{50}$) of $Ni^{2+}$ and Tebuconazole were 4.2, 39.1, respectively. The teratogenic index (TI) were 61.4 in $Ni^{2+}$ and 3.1 in Tebuconazole, respectively. These results reveals that $Ni^{2+}$ and Tebuconazole suppress the development of Black-spotted pond frog embryos at the low concentration as showing teratogenic effects in other assay system. Therefore, teratogen assay system using the Rana nigromaculata embryos could be useful as a tool to evaluate the toxicity of the pollutants in environment.

• Journal of Life Science
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• v.9 no.3
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• pp.333-340
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• 1999

The toxic effect of TBTO on Chlorella and Rotifer was observed. The value of 48hr-LC50 for Chlorella (3.3$\mu\textrm{g}$/L) estimated to be almost 500 times as high as that for Rotifer (6.7ng/L). A fertilized egg of olive flounder exposed in an embryo-formation stage was mostly influenced by TBTO toxicity when the fertilized egg at each stage until hatching was exposed to TBTO at the concentrations of 5 to 200ng/L. The values of LT50 were estimated to be 68.0, 41.0, 21.0, 13.0, 7.7 and 4.7 hours at 5, 10, 25, 50, 100 and 200ng/L of TBTO, respectively when the fertilized egg in a morula stage was exposed to TBTO, and the 48hr-LC50 was 8 ng/L. In case of TBTO treatment in an embryo-formation stage, the values of LT50 were 33.0, 12.5, 3.5, 1.3, 0.5 and 0.2 hours at 5, 10, 25, 50, 100 and 200ng/L of TBTO, respectively, and the value of 48hr-LC50 was 4ng/L. The values of LT50 were estimated to be 17.0, 11.0, 6.2, 4.0, 2.6 and 1.7 hours at 5, 10, 25, 50, 100 and 200ng/L of TBTO, respectively when the fertilized egg in a stage just before hatching was exposed to TBTO, and the 48hr-LC50 was below 1ng/L. The percentages of hatching were 46.2, 20.6, 21.9, 20.6 and 13.2% at 25, 50, 100, 250 and 500ng/L of TBTO, respectively when the fertilized egg in the stage just before hatching was exposed to TBTO and the measurement was done at second day after the completion of hatching. However no survival after the completion of hatching was found in all cases. With the treatment of 1, 5 and 10ng/L of TBTO, the percentages of hatching were 80.5, 70.0 and 44.1%, respectively. The percentages of survival until second day after the completion of hatching were 80.0, 63.3 and 9.1%, respectively. The percentages of hatching and survivability after the completion of hatching for the control were 84.5 and 82.5%, respectively.

• Environmental Analysis Health and Toxicology
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• v.18 no.4
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• pp.277-285
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• 2003

연안의 방조제 건설은 하천수와 바다물의 교환을 인위적으로 차단하고, 방조제 내부에는 호수를 형성하게 된다. 이렇게 형성된 인공호수의 수질은 호수로 유입되는 하천의 수질에 크게 영향을 받을 수 있으며, 인공호수의 수질이 악화되면 인근 해양생태계에 영향을 미칠 수 있다. 본 연구는 인공호수의 물이 바다로 방류될 때 해양생태계에 어떠한 영향을 미칠 것인지를 예측하고자 수행되었다. 방조제 건설로 이미 형성되어 있는 시화호, 아산호, 부사호와, 현재 건설중인 새만금 방조제로 인하여 형성될 호수의 유입 하천인 만경강에서 표층수를 채수하여 둥근성게(Strongylocentrotus nudus)의 정자와 수정란을 이용한 생물 검정을 실시하였다. 담수호의 수질을 해양생물을 이용하여 평가할 수 있도록 하기 위하여 시료의 염분을 조절하면서 실험을 실시하였다. 정자를 이용한 수정 실험 결과, 시화호에서는 수정률에 영향을 미치지 않았고, 아산호와 부사호에서는 약간의 영향이 있었으나 그 정도는 미약하였다. 반면, 만경강의 경우 수정률의 저해가 상당히 크게 나타났다. 수정란을 이용한 발생 실험 결과, 시화호, 아산호, 부사호에서는 약간의 영향이 나타난 반면, 만경강의 경우는 매우 큰 영향이 나타났다. 이들 실험 결과를 종합하면, 시화호, 아산호, 부사호의 수질은 대체로 양호하나, 만경강의 수질은 크게 우려되는 수준이었다. 따라서, 앞으로 건설될 새만금 호수와 인근 해양생태계를 보전하기 위해서는 만경강의 수질 개선을 위한 대책이 마련되어야 할 것이다.

• Korean Journal of Environmental Biology
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• v.39 no.3
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• pp.311-318
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• 2021

In this experiment, we investigated the toxicity of tebuconazole (fungicide) using domestic frog embryos, along the FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol. Bufo gargarizans, Hyla japonica, and Pelophylax nigromaculatus embryos were incubated, and investigation of the tebuconazole effect was performed by the probit analysis. As a result, depending on the concentrations of tebuconazole, the mortality and malformation rates were increased and larval body length was decreased. The teratogenic concentrations (EC₅₀) of tebuconazole were 34.4mg L^-1, 10.6mg L^-1, and 14.9mg L^-1, respectively, and the embryo lethal concentrations(LC₅₀) of tebuconazole were 74.7 mg L^-1, 38.5 mg L^-1, and 39.1 mg L^-1, respectively. The teratogenic index (TI) valuesof tebuconazole were 2.19, 3.58, and 2.65; thus, it showed teratogenicity in embryonic development of these three frogs. These results revealed that in this experiment, tebuconazole suppressed the development of embryos at a relatively low concentration. In addition, mortality, malformation ratios, malformation patterns, and growth rates were similar to the results from the other assay systems. Therefore, tebuconazole was thought to have an effect on the embryo development of domestic frogs. In future, it will be necessary to identify species specificity in order to the clarify the causes of differences in mortality, malformation rate, and malformation patterns depending on the species.

• Journal of Pharmacopuncture
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• v.19 no.4
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• pp.319-328
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• 2016

Objectives: Advancements in nanotechnology have led to nanoparticle (NP) use in various fields of medicine. Although the potential of NPs is promising, the lack of documented evidence on the toxicological effects of NPs is concerning. A few studies have documented that homeopathy uses NPs. Unfortunately, very few sound scientific studies have explored the toxic effects of homeopathic drugs. Citing this lack of high-quality scientific evidence, regulatory agencies have been reluctant to endorse homeopathic treatment as an alternative or adjunct treatment. This study aimed to enhance our insight into the impact of commercially-available homeopathic drugs, to study the presence of NPs in those drugs and any deleterious effects they might have, and to determine the distribution pattern of NPs in zebrafish embryos (Danio rerio). Methods: Homeopathic dilutions were studied using high-resolution transmission electron microscopy with selected area electron diffraction (SAED). For the toxicity assessment on Zebrafish, embryos were exposed to a test solution from 4 - 6 hours post-fertilization, and embryos/larvae were assessed up to 5 days post-fertilization (dpf ) for viability and morphology. Toxicity was recorded in terms of mortality, hatching delay, phenotypic defects and metal accumulation. Around 5 dpf was found to be the optimum developmental stage for evaluation. Results: The present study aimed to conclusively prove the presence of NPs in all high dilutions of homeopathic drugs. Embryonic zebrafish were exposed to three homeopathic drugs with two potencies (30CH, 200CH) during early embryogenesis. The resulting morphological and cellular responses were observed. Exposure to these potencies produced no visibly significant malformations, pericardial edema, and mortality and no necrotic and apoptotic cellular death. Conclusion: Our findings clearly demonstrate that no toxic effects were observed for these three homeopathic drugs at the potencies and exposure times used in this study. The embryonic zebrafish model is recommended as a well-established method for rapidly assessing the toxicity of homeopathic drugs.