• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1112-1117
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• 2008

Purpose : We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. Methods : The hippocampus of 7-day-old rats was cut into $350{\mu}m$ slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into $15{\mu}m$ thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. Results : 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P<0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. Conclusion : The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.

• Parasites, Hosts and Diseases
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• v.27 no.2
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• pp.87-100
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• 1989

Eimeria tenella, an intracellular protozoan parasite infecting the epithelial cells of the ceca of chickens, causes severe diarrhea and bleeding that can lead its host to death. It is of interest that 2. tenezla first penetrate into the mucosal intraepithelial Iymphocytes (IEL) before they parasitize crypt or villous epithelial cells. This in vitro study was undertaken to know whether the penetration of E. tenella into such a lymphoid cell is a beneficial step for the parasite survival and development. Three sequential experiments were performed. First, the in vitro established bovine kidney cell line, MDBK cells, were evaluated for use as host cells for E. tenella, through morphological observation. Second, the degree of parasite development and multiplication in MDBK cells was quantitatively assayed using radioisotope labelled uracil ($^3H-uracil$) . Third, the E. tenella sporozoites viability was assayed after preincubation of them with thicken spleen cells. E. tenella oocysts obtained from the ceca of the infected chickens were used for the source of the sporozoites. Spleen cells (I) obtained from normal chickens (FP strain) were preincubated with the sporozoites (T) at the E:T ratio of 100:1, 50:1 or 25:1 for 4 or 12 hours, and then the mixture was inoculated into the MDBK cell monolayer. Morphologically the infected MDBK cells revealed active schisogonic cycle of E. tenella in 3~4 days, which was characterized by the appearance of trophozoites, and immature and mature schizonts containing merogoites. The 3H-uracil uptake by E. tenella increased gradually in the MDBK cells, which made a plateau after 48~60 hours, and decreased thereafter. The uptake amount of $^3H-uracil$ depended not only upon the inoculum sixte of the sporozoites but also on the degree of time delay (preincubation; sporozoites only) from excystation to inoculation into MDBK cells. The 3H-uracil uptake became lower as the preincubation time was prolonged. In comparison, after preincubation of sporozoites with spleen cells for 4 or 12 hours, the 3H-uracil uptake was significantly increased compared with that of control group. From the results, it was inferred that, although the penetration of E. tenella sporozoites into the lymphoid cells such as IEL is not an essential step, it should be at least a beneficial one for the survival and development of sporozoites in the chicken intestine.

• Journal of Preventive Medicine and Public Health
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• v.29 no.3 s.54
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• pp.451-469
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• 1996

To identify the drug abuse status and its determinant factors in high school boys in Taegu, the study was performed from April to May, 1995. Study population were selected by cluster sampling method and total 5,665 students replied to the self-administered questionnaire survey (2,207 in academic high school, 3,458 in business high school). The major findings were as follows; The proportion of drinking, smoking experience was 55.0%, 45.8%, respectively, and the proportion of current drinker, current smoker was 27.2%, 27.5%. The drinking, smoking experience rate of second grade students was higher than first grade and it was higher in business high school boys. The proportion of a stimulant, a hallucinogen, hemp leaf cigarets experience was 3.2%, 1.6%, 0.1%, respectively. Drug abuse had significant association with home environment(lower economic status, frequent move, death of father or mother, apart from family), parents environment(parents' indifference, parents' drinking and smoking, etc.), school life(lower school grades, intimate friend's drug abuse, etc.), generous attitude to drug abuse, higher level of stress. Students who replied that the law prohibited immature person(students) from drinking and smoking showed lower drug abuse rate. In multiple logistic regression analysis, second grade students, business high school students, parents' indifference, lower school grades, intimate friend's drug abuse, no recognition of the fact that the law prohibits high school students from drinking and smoking, generous attitude to drug abuse, higher level of stress were significantly related with alcohol abuse and smoking. Other drugs abuse were related with above factors. On consideration of above findings, to prevent students from drug abuse, we have to try together in house, school, and society.

• Development and Reproduction
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• v.15 no.2
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• pp.179-185
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• 2011

Some phytoestrogens in soy and red wine, for example, might have beneficiary rather than adverse effects. In particular, dietary soy intake seems to be highly correlated with protection of breast cancer, osteoporosis and cardiovascular disorders. However, questions persist on the potential adverse effects of the main soy constituent genistein (GS) on female reproductive physiology. Previously we found that prepubertal exposure to GS could activate the reproductive system of immature female rats leading to precocious puberty onset, and intracerebroventricularly (ICV) injected GS could directly activate hypothalamic kisspeptin-GnRH neuronal circuits in adult female rats. The present study was performed to examine the hypothalamus-specific GS effects in prepubertal female rats and which subtype of estrogen receptor is mediated in this GS effect. Prepubertal female rats (PND 30) were anaesthetized, treated with single dose of GS (3.4 ${\mu}g$/animal), and sacrificed at 2 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly lowered the transcriptional activities of mTOR (1:$0.361{\pm}0.058$ AU, p<0.001) but increased that of GAD67 (1:$1.285{\pm}0.099$ AU, p<0.05), which are known to act as an upstream modulator of kisspeptin and GnRH neuronal activities in the hypothalamus, respectively. GS administration enhanced significantly the mRNA levels of KiSS-1(1:$1.458{\pm}0.078$ AU, p<0.001), and exerted no effect on the mRNA level of kisspeptin receptor GPR-54 (1:$1.29{\pm}0.08$ AU). GnRH gene expression was significantly decreased in GS-treated group compared to control group (1:$0.379{\pm}0.196$ AU, p<0.05). There was no difference in the mRNA level of $ER{\alpha}$ in the GS-treated group compare to control group (1:$1.180{\pm}0.390$ AU, Fig. 3A). However, icv infusion of GS significantly increased the transcriptional activities of $ER{\beta}$ (1:$4.209{\pm}0.796$ AU, p<0.01, Fig. 3B). Taken together, the present study indicated that the acute exposure to GS could directly alter the hypothalamic GnRH modulating system in prepubertal female rats. Our study strongly suggested the involvement of $ER{\beta}$ pathway in GS's hypothalamus-specific action, and this idea is consistent with the GS's well-known $ER{\beta}$-mediated protective action in breast cancer.

• Development and Reproduction
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• v.14 no.4
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• pp.281-286
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• 2010

Recent evidence has revealed that the intratesticular injection of hypertonic saline(20%) resulted in a chemically castrated state such as nadir testosterone levels in rats. To confirm the efficacy of this simple saline-injection method further, we investigated the changes in the gross and microscopic anatomy of testis. Our study comprised three groups; intact(control) group, orchidectomy group and saline-injection (experimental) group. Single dose of hypertonic saline (sterilized, $750{\mu}{\ell}/testis$) were directly administered into both testis of adult rats (about 300 g BW). Bilateral orchidectomy was performed at the same day of saline injection. Following 30 days post-injection, reproductive tissues were surgically removed, weighed and fixed for histological examination. The body weights were not changed in both orchidectomy group and saline-injection group when compared to those in intact group. The wet weights of testis were significantly decreased in saline-injection group when compared to those in intact group. The wet weights of epididymis and seminal vesicle and prostate were significantly decreased in orchidectomy group and saline-injection group when compared to those in intact group. Macroscopically, the testes exerted slight atrophy and the tunica albuginea seemed to be intact in saline injection group. Histologically, however, larger parts of testicular tissue underwent necrosis and were barely recognizable after hematoxylin-eosin staining. In the same section, only the opposite part of the injection site was stained showing abnormal state of cell layers mostly fibrosis and infiltrated leukocytes. Sloughing of immature germ cells from the basement membrane along with shedding cells in the intraluminal space was notable in most seminiferous tubules from the saline injected testis. The present study confirmed that the direct injection of hypertonic saline into testis can induce a castration-like, testosterone-depriving effects on accessory sex organs. Our findings suggest that the efficacy of this less expensive and minimally invasive method seems to be almost even with that of conventional orchidectomy and chemical castration, though more in-depth evaluation should be supported.

• Applied Microscopy
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• v.35 no.1
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• pp.41-56
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• 2005

Present study was performed to observe the tegumental ultrastructures by the developmental stages which derived from the experimental life cycle of Spirometra erinacei in laboratory conditions. In SEM view, coracidium was spherical in shape with numerous cilia, and its surface was covered with long cilia, tuberclelike projections with millet-like processes, and small holes. The body surface of procercoid was covered with numerous pointed microtriches except that of frontal pit with stout spine-like ones. However that of cercomer was covered with somewhat sparse blunt-tiped microtriches. Plerocercoids of 3 days old resembled the mature procercoid in shape, and their frontal pits were covered with numerous stout spine-like microtriches. However frontal pit and body surface in more than 5 days old ones were covered with conoid microtriches. On the surface of adult scolex, hairly long filamentous and stout short microtriches were mixedly distributed. Filamentous microtriches were more densely distributed in the anterior portion than in the posterior of scolex. The neck and immature proglottid were covered with only stout short conoid microtriches. In TEM view of coracidia, embryophore and oncosphere were obviously distinguished. The embryophore contained numerous glycogen particles, mitochondria and lipid granules. The cilia on the surface of embryophore rooted in the coracidial sheath, and consisted of 9 pairs of microtubules and 2 core complex. The oncosphere was covered with a thin and unarmed tegument, and was multi-nucleated. The protoplasmic layer of procercoid and plerocercoid consisted of disc-shaped bodies, vacuoles and mitochondria. Their tegumental cells commonly retained a nucleus, granular endoplasmic reticulums and secretory granules. The protoplasmic layer of plerocercoid was more compacted than that of procercoid. From the above results, it was confirmed that the tegumental ultrastructures are something different according to the developmental stages of S. erinacei.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.282-290
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• 1997

Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.

• Korean Journal of Soil Science and Fertilizer
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• v.40 no.4
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• pp.269-273
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• 2007

Occurrence of malodor could cause adverse impacts on human health and increase public interest. Therefore, scientific methods to decrease odor is required. Endeavor to decrease odor from compost however has not fully been successful. The purpose of this research is assessment of some amendments to reduce $NH_3$ from immature composts. Calcium hydroxide was applied to composts due to it's characteristics to increase pH. Activated carbon and zerovalent iron (ZVI) were selected because of their adsorption properties. The research results were as follows: Calcium hydroxide, activated carbon, zerovalent iron increased the composting temperature above $60^{\circ}C$. The addition of calcium hydroxide, activated carbon, and ZVI to compastry process increased pH 8.6 - 8.8 from $1^{st}$ day to $14^{th}$ day. During the 14 days of composting, addition of calcium hydroxide, activated carbon and ZVI changed EC from $2.15-0.66dS\;m^{-1}$, $1.48-1.11dS\;m^{-1}$, respectively and $1.77-0.68dS\;m^{-1}$. The difference in EC of the compost was due to irregularities of samples. Organic matter in the compost decreased through out theexcept control. The $NH_4-N/NO_3-N$ ratio of all experimental compost increased through the process. The addition of activated carbon, calcium hydroxide and ZVI decreased $NH_3$ from 0.1ppm, 0.7ppm and 1.7ppm more than the control (pig manure and sawdust), 9.3ppm, in 30 days of composting. In conclusion, odor from prematured compost decreased by addition of chemicals like calcium hydroxide, activated carbon, zerovalent iron. Moreover, use of these $NH_3$ reducers alone or together combined at different periods of composting etc. could decrease $NH_3$.

• Food Science and Preservation
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• v.23 no.2
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• pp.174-179
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• 2016

This study was to prepared vinegars using three kinds of pear cultivar with different maturities, 'Wonhwang', 'Niitaka', and 'Chuhwangbae', and investigate their physicochemical characteristics and antioxidant activities. Result showed that the firmness of 'Wonhwang' was much lower than other pear cultivars. No significant difference in soluble solid content was found among three pear cultivars. The total acidity of 'Chuhwangbae' pear vinegar was 4.89~7.98%, which was higher than that those of 'Wonhwang' and 'Niitaka' vinegars. The color of 'Niitaka' vinegar had a lower lightness and redness values, but a higher yellowness value, compared with the other two vinegars. The free sugar contents were mainly composed of fructose, glucose, and sorbitol, with the greatest content of sorbitol in all three peer vinegars. The total phenolics and flavonoids contents were 35.2~55.3 and 8.4~14.4 mg/100 mL, respectively. Antioxidant activity had a positive correlation with total phenolic contents. The DPPH free radical scavenging and $ABTS^+$ radical scavenging activity were 45.0~62.1% and 73.8~78.2%, respectively. From these results, we confirmed that immature pear cultivars such as 'Wonhwang', 'Niitaka' and 'Chuhwangbae' could be used as a raw materials to prepare vinegar regardless of maturity.

• Korean journal of applied entomology
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• v.53 no.4
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• pp.355-365
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• 2014

It has been unclear whether the diamondback moth, Plutella xylostella can overwinter in Korean field conditions. This study determined overwintering conditions of P. xylostella by conducting field exposure tests based on its cold tolerance and monitoring overwintering populations by direct examination of overwintering larval habitats and capturing adults with sex pheromone traps. In addition, the overwintering populations were analyzed using polymorphic genetic markers to trace their sources. When all immature stages of P. xylostella were exposed to $-5^{\circ}C$, which was the temperature much above their supercooling points, they significantly suffered with direct cold injuries, where larval stage was most tolerant to the cold injury. However, the exposure to $5^{\circ}C$ for a long period (4 weeks) did not give any significant cold injury to nonfeeding stages, while this treatment gave lethality to larval stage without diet. When all developmental stages of P. xylostella were exposed to open field conditions during winter, they exhibited significant decreases of survival rates. However, some protected and indoor conditions reduced the cold injuries and the diet provision significantly increased larval survival rates. Adult monitoring with sex pheromone during winter period indicated that the first captures were observed at similar periods at different locations (${\approx}260$ Km apart). The overwintering adults were captured until early April. Genetic variation of these overwintering populations was analyzed with polymorphic molecular markers, indicating significant genetic divergences among the overwintering populations. This study indicates that P. xylostella can overwinter in southern Korean fields or some protected greenhouses with host plants.