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Teratogenic and developmental toxic efects of etridiazole on zebrafsh (Danio rerio) embryos
Vasamsetti Bala Murali Krishna,Kim Nam-Seok,Chon Kyongmi,Park Hong-Hyun 한국응용생명화학회 2020 Applied Biological Chemistry (Appl Biol Chem) Vol.63 No.6
Etridiazole (EDZ), a thiadiazole-containing toxic chemical, is widely used as a fungicide. Regular usage of EDZ may reach and contaminate water bodies, but its adverse efects on aquatic vertebrates have not been well studied. Therefore, the present study aimed to evaluate the harmful efects of EDZ using zebrafsh (ZF) (Danio rerio) embryos. ZF embryos were treated with 3.75, 7.5, 15, 30, and 60 mg/L of EDZ. Subsequently, mortality and developmental toxicities were quantifed at 24, 48, 72, and 96 h post fertilization (hpf ). The results showed that embryo mortality was concentration- and time-dependent. The median lethal concentration (LC50) of EDZ at 96-h was 25.58±1.49 mg/L. Besides, EDZ induced a series of morphological deformities, including abnormal somite formation, abnormal eye pigmentation, abnormal tail morphology, tail kinks, skeletal malformations (lordosis, kyphosis, and scoliosis), and yolk sac edema in a concentration-dependent manner. Among the deformities, the most signifcant were reduced heartbeat and increased incidence of pericardial edema. The median efective concentration (EC50) of EDZ at 96-h was 17.93±2.22 mg/L and the 96-h teratogenic index (TI) value was 1.52. Taken together, these results indicate that EDZ is a teratogen, and primarily afects the cardiovascular system of ZF.
VIG-1 is required for maintenance of genome stability in Caenorhabditis elegans
Bala Murali Krishna Vasamsetti,박양서,조남정 한국통합생물학회 2018 Animal cells and systems Vol.22 No.3
To explore the function of VIG-1 in Caenorhabditis elegans, we analyzed the phenotypes of two vig-1 deletion mutants: vig-1(tm3383) and vig-1(ok2536). Both vig-1 mutants exhibited phenotypes associated with genome instability, such as a high incidence of males (Him) and increased embryonic lethality. These phenotypes became more evident in succeeding generations, implying that the germline of vig-1 accumulates DNA damage over generations. To examine whether vig-1 causes a defect in the DNA damage response, we treated worms with UV or camptothecin, a specific topoisomerase I inhibitor. We observed that the embryonic survival of the vig-1 mutants was reduced compared with that of the wild-type worms. Our results thus suggest that VIG-1 is required for maintaining genome stability in response to endogenous and exogenous genotoxic stresses.
Effects of carbendazim on normal growth and cardiac function of zebrafish embryos
Bala Murali Krishna Vasamsetti,Juyeong Kim,Kyongmi Chon,Jin-A Oh,Chang-Young Yoon,Hong-Hyun Park 한국농약과학회 2021 한국농약과학회 학술발표대회 논문집 Vol.2021 No.11
Carbendazim (CBZ) is a benzimidazole fungicide with a broad spectrum of applications in agriculture, horticulture, forest management, and home gardening. The toxic effects of CBZ on mammals have been well documented. However, the adverse effects of CBZ on aquatic organisms are not completely understood. The current study aimed to investigate the developmental toxic effects of CBZ on zebrafish (ZF) embryos. The developmental toxicity test was performed in accordance with the OECD 236 test guidelines, at 0.40, 0.48, 0.58, 0.69, and 0.83mg/L of CBZ. In parallel, DMSO (0.03%)-treated embryos were maintained as a control. The embryo mortalities and developmental deformities were scored at 24, 48, 72 and 96 hours post fertilization (hpf). The median lethal concentration (LC50) of CBZ at 96-h was 0.77 mg/L. In addition to the mortalities, CBZ induced several abnormalities. CBZ induced malformed somites (50.2%), curved spines (75.7%), reduced eye pigment (60.7%), yolksac edema (13.6%), and swimming disability (82.86%) at a dose of 0.69 mg/L. The median effective concentration (EC50) of CBZ for malformed somites, reduced eye pigment, and pericardial edema were 0.77, 0.64 and 0.66mg/l, respectively. The average heartbeats of the control group were 169.8 ± 2.7 per minute, whereas the average heartbeats of the CBZ-exposed groups ranged from 167 ± 6.8 (0.40mg/L) to 88.0 ± 10.1 (0.83mg/L) per minute, indicating that CBZ impairs cardiac function. CBZ-treated ZF embryos also exhibited heart-related abnormalities such as pericardial edema (18.15 - 80.00%), hyperemia (7.5 - 100%), and irregular blood flow (2.5 - 47.1%), implying that CBZ affects cardiac development. Overall, these findings suggest that CBZ is teratogenic to zebrafish and affects the cardiac function in the earlier developmental stages.
Bala Murali Krishna Vasamsetti,In-sul Hwang,Heasun Lee,Sung-June Byun,Hyeon Yang,Sun A Ock,Hwi-Cheul Lee,Jae-Seok Woo,Keon Bong Oh 한국수정란이식학회 2018 한국수정란이식학회 학술대회 Vol.2018 No.11
In our previous studies, the cardiac xenotransplantation from an alpha-1,3-galactosyltransferase knockout pig (GT-MCP-MCP) to cynomolgus monkeys showed a mean survival of 38 days. The objective of this study is to genetically upgrade the GT-MCP-MCP pig, to further enhance membrane cofactor protein (MCP) expression and to express an endothelial specific thrombomodulin (TBM). MCP is a complement regulatory protein and TBM is a coagulation inhibitor. As the dicistronic cassette for wild-type-based MCP and TBM concurrent expressions does not show any increase of MCP, we optimized the MCP codon usage (mMCP) and substituted mMCP for MCP. When the mMCP-TBM cassette was transfected to HeLa cells, we were able to find an increased expression of MCP and endothelial cell-specific TBM expression. The cassette was then transfected into ear-skin fibroblasts isolated from one-month-old #23-4 of a GT-MCP-MCP pig, and the cell populations expressing MCP were obtained by MACS cell sorting. We performed a single cell culture of the selected cells, and obtained clones over expressing 90% MCP. The cells of a clone were used as a donor for nuclear transfer and generated GT-MCP/-MCP/mMCP/TBM pig. The transgenic pig was confirmed to be carrying the cells expressing MCP and functioning as an inhibitor against the cytotoxic effect of normal monkey serum, comparable with donor cells. Thus, we believe that the GT-MCP/-MCP/mMCP/TBM transgenic pig would be potential for the prolongation of xenograft survival in the recipients.