• Development and Reproduction
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• v.11 no.3
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• pp.263-272
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• 2007

Contrast to mouse where its in vitro maturation rates are high without specific supplements or presence of the cumulus cells, there are some species, such as porcine, where its in vitro oocyte maturation rates are still very low. This comparative study was conducted to investigate the role of malate dehydrogenase(Mor2) during oocyte maturation by RNAi in the mouse and porcine. The Mor2 double-stranded RNA(dsRNA) was prepared speciesspecifically and microinjected into the cytoplasm of denuded germinal vesicle(GV) oocytes. Oocytes were cultured for 48 h(porcine) and 16 h(mouse) in M199 with 10% porcine follicular fluid, pyruvate, p-FSH, EGF, cystein, and estradiol-$17{\beta}$. We measured changes in oocyte morphology, maturation rates and mRNA levels after Mor2 RNAi. We confirmed gene sequence-specific knock down of Mor2 mRNA in both species after Mor2 RNAi. In contrast to our previous finding that mMor2 RNAi resulted in GV arrest in the mouse, we found that pMor2 RNAi resulted in MI arrest in denuded porcine oocytes(58%), but developed to MII(84.4%) in COCs. To determine whether this difference between mouse and porcine RNAi is due to differences in culture media, we cultured mouse oocytes in the M199 media for 16 h after mMor2 RNAi. Mouse oocytes were developed to MII stage(62%) and there was no statistical difference compared to that of non-injected(76.8%) and buffer-injected(73.3%) control groups. Therefore, we concluded that the mouse and porcine oocytes are having different metabolic systems in relation to malate dehydrogenase for oocyte maturation. This could be a basis for differences in maturation rates in vitro in two species. Further scrutinized studies on the metabolic pathways would led us in finding better culture system to improve oocyte maturation rates in vitro, especially in more challenging species like the porcine.

• Development and Reproduction
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• v.11 no.3
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• pp.155-165
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• 2007

Gonad index (GI), conditon index, the reproductive cycle with gonadal development of the Neptunea (Barbitonia) arthritica cumingii, were investigated histologically, based on the samples which have been collected from the subtidal zone of Oeyeondo, Boryeong, Korea from January to December, 2006. Neptunea (Barbitonia) arthritica cumingii is dioecious and oviparous. Monthly changes in the gonad index (GI), studied for determination of spawning period, were closely associated with gonad developmental phases. The GI reached a maximum in April, and gradually decreased from May to August due to spawning. The gonadsomatic index and condition index showed similar patterns to gonad developmental phases and the spawning period. The reproductive cycle according to gonad developmental phases of this species can be classified into five successive stages in females and males: in females, early active stage (September to October), late active stage (November to February), ripe stage (February to June), partially spawned stage (May to August) and recovery stage (June to August); in males, the early active stage (September to October), late active stage (November to February), ripe stage (February to June), copulation (April to July), and recovery stage (July to August). Spawning occurred between May to August in females and April to July in males, and spawning peak in females was observed between June and July when the seawater temperature rose to above $19^{\circ}C$. Percentages of first sexual maturity of female and male snails ranging from $50.1{\sim}60.0\;mm$ in shell height were over 50%, and 100% for snails over 60.1 mm in shell height. The sex ratios of females to males were not significantly different from a 1:1 sex ratio.

• Development and Reproduction
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• v.11 no.3
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• pp.167-177
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• 2007

In the present study, we isolated three human adult stem cells including adipose tissue-derived stem cells(HAD), umbilical cord-derived stem cells(HUC), and amnion-derived stem cells(HAM) and analysed their characteristics. And we examined whether HAD could be used as therapeutical cells for the heart diseases. Both HAM and HUC appeared very similar morphology but HAD was different. Doubling time of HUC was most fast, but total doubling numbers of HUC was same with HAM. Total doubling numbers of HAD was much more than others. Expression patterns of genes and proteins of three human adult stem cells were very similar. Also they were differentiated into adipocytes, osteocytes, and chondrocytes. In addition, they expressed many cardiomyocyte-related genes. But expression pattern of genes is a little different. When HAD were cultivated in the presence or absence of various combinations of BMP and FGF after 5-azacytidine expose for 24 h, expression of Cmlc-1, and ${\alpha}1c$ genes was significantly increased. However, expression of troponin T, troponin I and Kv4.3 genes was not changed. Based on these observations, it is suggested that HAD, HUC, and HAM might be used as potentially therapeutical cells for clinical application.

• Development and Reproduction
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• v.11 no.3
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• pp.187-193
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• 2007

Ethane 1,2-dimethane sulfonate(EDS), a Leydig cell specific toxicant, has been widely used to create the reversible testosterone withdrawal rat model. Though the maintenance of epididymal structure and function is highly dependent on the testosterone secreted from testis, its derivatives, dihydroxytestosterone(DHT) and estrogen, might have crucial roles. The aim of present study was to monitor the expression patterns of sex steroid receptors, cytochrome P450 aromatase(P450arom) and $5{\alpha}$-reductase in the rat epididymis up to 7 weeks after EDS injection. Adult male rats($350{\sim}400g$) were injected with a single does of EDS(75 mg/kg i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. The transcriptional activities of the target genes were evaluated by semi-quantitative RT-PCRs. The transcript level of estrogen receptor alpha($ER{\alpha}$) in EDS group was significantly higher than control level on week 1(P<0.01). After week 2, there was no significant difference in $ER{\alpha}$ levels between EDS group and control. The transcript level of estrogen receptor beta($ER{\beta}$) in EDS group was significantly higher than control level on week 1(P<0.05), lowered on weeks 2 and 3(P<0.05 and P<0.01, respectively), fluctuated during weeks 4 and 6, and elevated on week 7(P<0.05). The androgen receptor (AR) message levels increased significantly week 2(P<0.01), then returned to control level on week 3. In contrast, expression of cytochrome P450 aromatase(P450arom) decreased sharply during weeks $1{\sim}3$(P<0.01 on weeks 1 and 2; P<0.05 on week 3), then went back to control level on week 4. The mRNA level of $5{\alpha}$-reductase type 2($5{\alpha}$-RT2) increased significantly on week 4(P<0.01), then returned to control level. The present study indicated that EDS administration could induce reversible alterations in the transcriptional activities of sex steroid hormone receptors and androgenconverting enzymes in rat epididymis. EDS injection model will be useful to clarify the regulation mechanism of mammalian epididymal physiology.

• Korean Journal of Environmental Biology
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• v.37 no.4
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• pp.625-639
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• 2019

Microplastics are one of the substances threatening the marine ecosystem. Here, we summarize the status of research on the effect of microplastics on marine life and suggest future research directions. Microplastics are synthetic polymeric compounds smaller than 5 mm and these materials released into the environment are not only physically small but do not decompose over time. Thus, they accumulate extensively on land, from the coast to the sea, and from the surface to the deep sea. Microplastic can be ingested and accumulated in marine life. Furthermore, the elution of chemicals added to plastic represents another risk. Microplastics accumulated in the ocean affect the growth, development, behavior, reproduction, and death of marine life. However, the properties of microplastics vary widely in size, material, shape, and other aspects and toxicity tests conducted on several properties of microplastics cannot represent the hazards of all other microplastics. It is necessary to evaluate the risks according to the types of microplastic, but due to their variety and the lack of uniformity in research results, it is difficult to compare and analyze the results of previous studies. Therefore, it is necessary to derive a standard test method to estimate the biological risk from different types of microplastics. In addition, while most of the previous studies were conducted mostly on spheres for the convenience of the experiments, they do not properly reflect the reality that fibers and fragments are the main forms of microplastics in the marine environment and in fish and shellfish. Furthermore, studies have been conducted on additives and POPs (persistent organic pollutants) in plastics, but little is known about their toxic effects on the body. The effects of microplastics on the marine ecosystems and humans could be identified in more detail if standard testing methods are developed, microplastics in the form of fibers and fragments rather than spheres are tested, and additives and POPs are analyzed. These investigations will allow us to identify the impact of microplastics on marine ecosystems and humans in more detail.

• Korean Journal of Environmental Biology
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• v.37 no.4
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• pp.568-578
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• 2019

The nymphal development of the potato aphid, Macrosiphum euphorbiae (Thomas), was studied at seven constant temperatures (12.5, 15.0, 17.5, 20.0, 22.5, 25.0, and 27.5±1℃), 65±5% relative humidity (RH), and 16:8 h light/dark photoperiods. The developmental investigation of M. euphorbiae was separated into two steps, the 1^st through 2^nd and the 3^rd through 4^th stages. The mortality was under 10% at six temperatures. However, it was 53.0% at 27.5℃. The developmental time of the entire nymph stage was 15.5 days at 15.0℃, 6.7 days at 25.0℃, and 9.7 days at 27.5℃. In the immature stage, the lower threshold temperature of the larvae was 2.6℃ and the thermal constant was 144.5 DD. In our analysis of the temperature-development experiment, the Logan-6 model equation was most appropriate for the non-linear regression models (r²=0.99). When the distribution completion model of each development stage of M. euphorbiae larvae was applied to the 2-parameter and 3-parameter Weibull functions, each of the model's goodness of fit was very similar (r²=0.92 and 0.93, respectively). The adult longevity decreased as the temperature increased but the total fecundity of the females at each temperature was highest at 20℃. The life table parameters were calculated using the whole lifespan periods of M. euphorbiae at the above six temperatures. The net reproduction rate (R₀) was highest at 20.0℃(63.2). The intrinsic rate of increase (r_m) was highest at 25℃(1.393). The finite rate of doubling time (D_t) was the shortest at 25.0℃(2.091). The finite rate of increase (λ) was also the highest at 25.0℃(1.393). The mean generation time(T) was the shortest at 25.0℃(9.929).

• Korean Journal of Environmental Biology
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• v.37 no.4
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• pp.749-758
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• 2019

The aim of this study is to provide a scientific basis for decision making regarding environmental damage in case of future chemical accidents by evaluating the ecotoxicity of 4 substances requiring preparation for accidents. For this purpose, acute and chronic toxicities of nitric acid, sulfuric acid, hydrogen peroxide, and ammonia solution, which can change the physical and chemical properties of soil to Paronychiurus kimi(Collembola) were investigated. The pH of artificial soil spiked with a series of test chemical concentrations was also measured. The pH of soil spiked with 10,000 mg kg^-1 of soil nitric acid, sulfuric acid, hydrogen peroxide, and ammonia solution were 2.86, 2.72, 7.18 and 9.69, respectively. The 28-d LC₅₀ of nitric acid, sulfuric acid, hydrogen peroxide and ammonia solution were 2,703, 5,414, 3,158 and 859 mg kg^-1 soil dry wt., respectively and 28-d EC₅₀ were 587, 2,148, 1,300 and 216 mg kg^-1 soil dry wt., respectively. These results indicated that the mortality and juvenile production of P. kimi were influenced by not only the soil pH but also by the reduced organic content and products produced by the reaction of soil with the tested chemicals. Given the fact that most substances requiring preparation for accidents can change soil characteristics, assessment and restoration methods that take into account changes in soil properties are needed for accurate decision making after chemical accidents.

• Journal of Embryo Transfer
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• v.21 no.1
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• pp.35-43
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• 2006

Ions play important roles in various cellular processes including fertilization and differentiation. However, it is little known whether how ions are regulated during early embryonic development in mammalian animals. In this study, we examined changes in $Ca^{2+}\;and\;K^+$ concentrations in embryos and oviduct during mouse early embryonic development using patch clamp technique and confocal laser scanning microscopy. The intracellular calcium concentration in each stage embryos did not markedly change. At 56h afier hCG injection when 8-cell embryos could be Isolated from oviduct, $K^+$ concentration in oviduct increased by 26% compared with that at 14h after injection of hCG During early embryonic development, membrane potential was depolarized (from -38 mV to -16 mV), and $Ca^{2+}$ currents decreased, indicating that some $K^+$ channel might control membrane potential in oocytes. To record the changes in membrane potential induced by influx of $Ca^{2+}$ in mouse oocytes, we applied 5 mM $Ca^{2+}$ to the bath solution. The membrane potential transiently hyperpolarized and then recovered. In order to classify $K^+$ channels that cause hyperpolarization, we first applied TEA and apamin, general $K^+$ channel blockers, to the bath solution. Interestingly, the hyperpolarization of membrane potential still appeared in oocytes pretreated with TEA and apamin. This result suggest that the $K^+$ channel that induces hyperpolarization could belong to another $K^+$ channel such as two-pore domain $K^+(K_{2P})$channel that a.e insensitive to TEA and apamin. From these results, we suggest that the changes in $Ca^{2+}\;and\;K^+$ concentrations play a critical role in cell proliferation, differentiation and reproduction as well as early embryonic development, and $K_{2P}$ channels could be involved in regulation of membrane potential in ovulated oocytes.

• Journal of fish pathology
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• v.18 no.1
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• pp.19-28
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• 2005

Scuticociliatosis has badly settled one of most damaging diseases during the seedling production process of olive flounder. Paralichthys olivaceus in Korea. We isolated a new type of Scuticociliate from flounder. The parasite metamorphoses to ciliate and cyst phases with each other by environmental changes and survive for a relatively long span. The ciliate was measured average 41.8 ${\mu}m$ in length and 21.0 ${\mu}m$ in width, and cyst was 17.0 ${\mu}m$ and 13.5 ${\mu}m$, respectively. Nutritional condition was determined as a major parameter of metamorphosing between ciliate and cyst stages. The ciliate transforms to a cyst stage because of food shortage, and the cyst returns to a ciliate stage with a favorite environmental condition and shows active growth and reproduction. The ciliate multiplied at the maximal density of $2.9 {\times} 10^5 {m\ell}^{-1}$cells in vitro cultivation at $15 ^{\circ}C$temperature using MS BHI medium and bacterial food sources. The ciliate could be proliferated at a 2.5 to $30 ^ \circ}C$ temperature range, pH 6 to 9, and 1 to 55 ppt salinity. Particularly, it survived over one week at $0 ^{\circ}C$temperature showing a high resistance against unfavorable environmental conditions. And the cyst survived for 320 days in the condition of $5 ^{\circ}C$with no feeding, but its survival period was markedly shortened in higher temperature conditions. The chemotherapeutants (formalin and hydrogen peroxide) were clarified as effective chemicals against the ciliate during in vitro trials, but the effect of therapeutants differed in proportion, depending upon the density and the bathing time of chemical compounds.

• Korean Journal of Ecology and Environment
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• v.49 no.1
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• pp.31-41
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• 2016

Deutzia paniculata is an endemic species to the Korean Peninsula. Despite of importance for conservation, the population structure and habitat characteristics of D. paniculata have not been determined yet. We analyzed the ecological characteristics of the species based on the literature review and field survey. Field survey was conducted on May to October 2014 during which 11 quadrats of size $15{\times}15m$ were studied in six regions. Each of the quadrats were further divided into $5{\times}5m$ small quadrats and population characteristics were recorded. The population and habitat characteristics were analyzed, including species abundance (density and coverage), demographic attributes (flowering rates and fruiting plants), vegetation (structure, species composition), light availability (transmitted light and canopy openness) and soil characteristics (temperature and humidity). We found that D. paniculata mainly distributed in Gyeongsangdo (including Taebaek in Gangwondo) along a broad elevational range of 290~959 m (mean: 493 m) above sea level. In preferred habitat the species grows within the slope range of $7^{\circ}$ and $35^{\circ}$ with the average of $16^{\circ}$. D. paniculata was generally distributed on talus deposits and low adjacent slopes. The average number of individual plants per small quadrat was 12.5 with the mean density $0.5stems\;m^{-2}$. The vegetative reproduction was frequent in D. paniculata and mean flowering rate was as low as 15%. Altogether 138 taxa were found in whole observation area with the dominant tree species mainly spring ephemerals, such as Cornus controversa (importance value: 25.5%) and Fraxinus rhynchophylla (importance value: 15.8%). Although, C. controversa usually grows on steep slopes and F. rhynchophylla mostly distributed at high-altitudes, however, both species distributed in disturbed environments and among talus deposits. Thus based on our results, we concluded that D. paniculata is a disturbance-prone species, primarily existing in habitats subjected to natural disturbances, such as floods. The species occurs less at anthropogenically disturbed sites, thus there is no apparent threat to the populations and habitat of D. paniculata.