• Reproductive and Developmental Biology
• /
• v.39 no.3
• /
• pp.77-81
• /
• 2015

This study was carried out to evaluate the effects of embryonic stage and toxicities of cryoprotectant on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology. Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and ethylene glycol (EG) were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or EG. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO ($68.1{\pm}24.1$) at the 2-cell stage was significantly lower than that were treated with EG ($81.2{\pm}27.0$) or the control ($99.0{\pm}18.3$) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the EG treated group (p<0.05). Both DMSO ($15.4{\pm}1.5$) and EG ($10.2{\pm}1.4$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.1{\pm}0.9$, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the EG treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or EG at room temperature. When comparing two cryoprotective agents, EG appeared to be less toxic than DMSO at least in a murine embryo model.

• Reproductive and Developmental Biology
• /
• v.32 no.3
• /
• pp.141-146
• /
• 2008

To search a new porcine pheromonal odorant, the models of four type (2D-QSAR, HQSAR, CoMFA & CoMSlA) were derived from quantitative structure-activity relationship (QSAR) between tetrahydrofuran-2-yl family compounds and their observed binding affinity constants (Obs.p$[Od]_{50}$). The optimized CoMFA model (predictability; $r^{2}_{cv.}(q^2)$=0.886 & correlation coefficient: $r^{2}_{ncv.}$=0.984) from ligand based approaches was confirmed as the best model among them. The $N^{1}$-allyl-$N^{2}$-(tetrahydrofuran-2-yl)methyl)oxalamide (P1), 2-(4-trimethylammoniummethylcyclohexyloxy)tetrahydrofurane (P5) and 2-(3-trimethylammoniummethylcyclohexyloxy)tetrahydrofurane (P6) molecules predicted as porcine pheromonal odorant by the CoMFA model were showed relatively high binding affinity constant values (Pred.p$[Od]_{50}=8{\sim}10$) and very lower toxicity values against some sorts of toxicity.

• Korean Journal of Ecology and Environment
• /
• v.41 no.2
• /
• pp.117-127
• /
• 2008

Green algae are important primary producers in aquatic ecosystem, and they are sensitive test species in bioassay. Green algae are broadly used to assess the adverse effects of various chemicals by measuring the inhibition of metabolism, reproduction and survival. In this study, we extensively gathered domestic and foreign toxicity test methods conducted using green algae, which are distributed in Korean water environment. Selected eight domestic green algae were Chlamydomonas reinhardtii, Desmodesmus subspicatus (=Scenedesmus subspicatus), Scenedesmus abundans, Scenedesmus acutus, Scenedesmus quadricauda, Podohedriella falcata (=Ankistrodesmus falcatus), Pseudokirchneriella subcapitata (=Selenastrum capricornutum), and Chlorella vulgaris. Forty four test methods were collected from the standard test ones, government reports, SCI papers and Korean research papers. P. subcapitata and D. subspicatus are the most common test species recommended by the standard test methods. Initial cell density and dilution water were the main differences among the test methods we collected. We proposed the suitable ecotoxicity test methods based on domestic green algae in Korea. This study could be a fundamental basis to establish the ecotoxicity test methods by green algae distributed in Korea.

• Development and Reproduction
• /
• v.3 no.1
• /
• pp.45-51
• /
• 1999

It is crucial to remove trophectoderm (TE) cells of blastocysts for an efficient isolation of pluripotent embryonic stem (ES)-like cells from bovine blastocysts. We evaluated the effectiveness of chemosurgery using calcium ionophore A23l87 (CIPA) by investigating the viability and pluripotency of ES-like cell lines isolated from in vitro-produced bovine blastocysts after CIPA treatment. The blastocysts treated with 50 $\mu$M CIPA for 25 min colonized most efficiently (51% of blastocysts) and developed to ES-like cell lines through 10 passages (4.8% of blastocysts) among CIPA-treated groups with different concentration and duration. In comparison with CIPA-untreated blastocysts, the colonization rate and overall viability of the CIPA-treated blastocysts were five times higher, suggesting that CIPA treatment condition defined in this study was highly efficient for establishing ES-like cell lines without apparent toxicity of CIPA. We evaluated in vitro pluripotency of the established three ES-like cell lines by examining alkaline phosphatase (AP) activity, capability of embryoid body formation, and chromosomal euploidity of the cells. Our cells showed a heterogeneous AP activity similarly to other reports. The cells were able to form simple embryoid bodies during suspension culture and majority of them showed a normal chromosome number of 60, the euploid chromosomal complement of bovine Therefore, our data suggest that CIPA treatment can be safely used for an efficient isolation of ES-like cell lines from bovine blastocysts.

• The Korean Journal of Pesticide Science
• /
• v.7 no.4
• /
• pp.280-287
• /
• 2003

To assess the risk of pesticides on earthworm, the acute toxicities of 10 pesticides were investigated and their toxicity exposure ratios(TERs) were calculated. As the TERs of paraquat dichloride and pendimethalin were more than 100, their risks were rated negligible. Risk of benfuracarb, cadusafos, chlorpyrifos-methyl, endosulfan, isazofos and parathion which have TERs of $10\sim100$ were rated low. However, risk of imidacloprid and phorate which have TER of less than 10 were estimated highly to need a reproduction study. Earthworms were exposed to twenty two pesticides including dazomet 98% GR having PECs of more than $5mg{\cdot}kg^{-1}$ in artificial soil at standard and double dose for 14 days. All the earthworms exposed to dazomet 98% GR and metam-sodium 25% SL were died to show their high risk, while no serious adverse effects were observed in the soil treated with 15 pesticides, calcite 95% WP, calcium polysulfide 36% CF, chlorothalonil 75% WP, daminozide 85% WP, dichlonil 6.7% GR, etridiazole 25% EC, fosetyl-Al 80% WP, glyphosate 41 % SL, hymexazol 30% SL, iprodione 50% WP, machine oil 95% EC, mancozeb 75% WP, propineb 70% WP, terbuthylazine 80% WP and triazophos 40% EC. In case of thiophanate-methyl 70% WP, copper hydroxide 77% WP, dimethoate 46% EC, tolclofos-methyl 50% WP and propamocarb hydrochloride 67% SL, any effect did not show clearly, suggesting an additional subchronic toxicity study. The risk of thiophanate-methyl 70% WP to earthworm was estimated high, considering its subchronic effect, while effects of copper hydroxide 77% WP, dimethoate 46% EC, tolclofos-methyl 50% WP and propamocarb hydrochloride 67% SL to earthworms were negligible, considering no adverse effects in subchronic tests.

• Journal of the Korean Society of Marine Environment & Safety
• /
• v.21 no.6
• /
• pp.655-661
• /
• 2015

This study intends to evaluate the effect of nitric acid($HNO_3$) spill accidents on the marine ecosystem, while $HNO_3$ is known as one of the typical HNS. For this purpose, we performed (1) the growth inhibition test by using phytoplankton(Skeletonema costatum), (2) acute and chronic toxicity test by using invertebrate(Brachionus plicatilis and Monocorphium acherusicum), (3) fish(Cyprinodon variegatus) and (4) bacteria(Vibrio fischeri). In these tests, we observed the (1) pH changes induced by the nitric acid spill and (2) changes in nitrate($NO_3$) concentration disassociated from nitric acid after the accident, respectively. The toxicity test result on pH changes induced by $HNO_3$ shows that the no observed effect concentration(NOEC), lowest observed effect concentration(LOEC) and 50 % effect concentration($72h-EC_{50}$) values of M. acherusicum are pH 7 (0.3 mM), pH 5(1.1 mM) and pH 5.2(1.4 mM), respectively, indicating that M. acherusicum is the most sensitive species. The chronic toxicity test (population growth rate test) on $NO_3{^-}$ of B. plicatilis show that the NOEC, LOEC and $96h-EC_{50}$ are 5.9 mM, 11.8 mM and 32.6 mM, respectively, indicating that B. plicatilis is the most sensitive species. In conclusion, toxic effecst on the marine organism caused by the nitric acid spill accident is determined to be so slightly except for the most adjacent area of the ship in pH scale and such concentration of nitrate, to the extent of directly influencing the survival and reproduction of the marine organism, is determined practically not to be applicable in the typical accidents in the sea.

• Journal of Environmental Health Sciences
• /
• v.39 no.5
• /
• pp.426-437
• /
• 2013

Objective: Gotjawal refers to a special geographical designation found in Jeju Province, Korea, where vegetation forms over a rocky area. Due to the important ecological value of Gotjawal, international concern about such areas is growing. However, only limited information is available regarding environmental contamination of Gotjawal. This study was conducted in order to investigate the levels of contamination and associated ecotoxicological effects of surface water bodies in Gotjawal. Methods: Surface water samples were collected at three sampling sites in two Gotjawal areas and were analyzed for several pharmaceuticals and pesticides in consideration of the potential sources of contamination. The ecotoxicity of the samples was measured using water fleas (Daphnia magna and Moina macrocopa) and zebrafish (Danio rerio). In addition, effects on the gene transcription of zebrafish were investigated following exposure to the samples. Results: Nine pharmaceuticals were detected in the samples, but none of the target pesticides were detected. Following acute exposure to two surface water samples, the survival of waterfleas was significantly reduced, but reproduction was not affected. In zebrafish, time-to-hatch was delayed and fry survival was decreased in some samples. On the other hand, at the transcriptional level, there were no genes significantly influenced by exposure to the samples. Conclusion: This is the first study investigating environmental contamination in Gotjawal areas and associated ecotoxicities. Further studies are warranted to identify the cause of acute Daphnia toxicity and to determine potential consequences of longer-term exposure in Gotjawal areas.

• Reproductive and Developmental Biology
• /
• v.38 no.3
• /
• pp.107-114
• /
• 2014

Oral exposure of humans by excess amounts of arsenic may cause disturbances of the reproductive system. In the present study, such exposure was modelled in rats, with the support of sperm principal parameters and histopathological observations. Male Sprague-Dawley rats were randomly divided into three groups where the group I was served as a normal control, group II was received sodium meta-arsenite as arsenic (10 mg/kg b.w/day) and a combination of sodium meta-arsenite and sodium selenite (3 mg/kg b.w/day) in group III. After 6 weeks, there was no significant change in testis weight and in total motility of all the three experimental groups, whereas, rapid moving spermatozoa, moderately moving spermatozoa and slow moving spermatozoa were significantly decreased in arsenic treated rats as compared to control rats. The other sperm principal parameters like progressiveness, average path velocity, straightness linear velocity (VSL), curvilinear velocity (VCL), straightness, linearity sperm head elongation ratio, area, linearity amplitude of lateral head department (ALH) and beat cross frequency (BCF) were found to be reduced in arsenic intoxicated rats. These results are not correlated with the histological studies. On oral administration of selenium ameliorated the adverse effects of arsenic as compared to arsenic alone treated rats. Our findings clearly demonstrate that administration of selenium could prevent some of the deleterious effects of arsenic in the testis.

• Journal of Environmental Health Sciences
• /
• v.29 no.2
• /
• pp.7-15
• /
• 2003

Di-2-ethylhexyl phthalate (DEHP), is a widely used plasticizer known to be a suspected endocrine disrupter, but its exact effects on aquatic organisms are not yet known. When Japanese medaka (Oryzias latipes) were exposed from the time of hatching to 3 months of age to an aqueous DEHP solution at nominal concentrations of 1, 10, and 50 $\mu\textrm{g}$/l, DEHP treated female fish showed distinct reproductive effect. And the midge (Chironomus riparius.). an aquatic invertebrate, was exposed to DEHP to evaluate the effects on reproductive processes via sediment toxicity. The test endpoints included emergence, sex ratio, fecundity, and the viability of F1 offspring egg ropes. The result implied that the normal developmental and/or reproductive processes in C. riparius had been disrupted when exposed to DEHP, the effect also being displayed in the next generation. In summary, DEHP hinders the development of reproductive organs in the female Japanese medaka and C. riparius.

• Development and Reproduction
• /
• v.15 no.1
• /
• pp.17-24
• /
• 2011

Lipid metabolites involved in cellular regulation as signaling mediators. Prostaglandins (PGs), metabolites of lipid are involved to pregnancy at the time of implantation but the functional roles of PGs on embryo development are still controversy and largely unknown. In previous report, the levels of $PGE_2$ and $PGF_{2a}$ at embryos of morula stage and blastocyst stage were explored (Cheon et al., 1998). In this study, the previous suggestion was confirmed and the possible downstream mediator of prostaglandin $E_2$ and prostaglandin $F_{2a}$ on the expansion and hatching of mouse embryo was examined. As expected, developmental rate of the blastocyst to expanded stage was a concentration-response curve that showed the highest expansion rate at 10 ${\mu}M$ $PGE_2$, but at 100 ${\mu}M$ $PGE_2$, the rate was decreased. In contrast to the $PGE_2$, $PGF_{2a}$ stimulated expansion without toxicity at highest concentration. Cotreatment of PGs with indomethacin overcame the inhibitory effects of indomethacin in expansion. Exogenous PGs also improved the development of expanded embryos to the hatching stage. Besides, PGs receptors' transcripts detected at blastocyst. $PGE_2$ was caused of calcium fluctuation in the blastocyst but $PGF_{2a}$ did not. The changes of intracellular calcium concentration were different between indomethacin pretreated embryos and non-treated embryos. Based on these results it is suggested that PGs work as paracrine and/or autocrine factors through calcium and the others which were not identified in this study.