• 한국환경농학회지
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• 제6권1호
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• pp.17-24
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• 1987

Benzene환(環)이 균일하게 표지된 제초제 $^{14}C-Bentazon$(3-isopropyl-2, 1, 3-benzothiadiazin-4-one-2, 2-dioxide)을 서독과 한국의 토양에 5.51 ㎎/㎏과 25.05㎎/㎏의 두 수준으로 처리하여 $23{\pm}1^{\circ}C$에서 105일간 호기적으로 배양했을때 두 토양에서 모두 $^{14}CO_2$로 분해되는 속도는 주당 0.6%와 0.2%이었다. Bentazon을 처리한 토양을 용매로 추출한 결과 증류수가 가장 우수한 용매임이 밝혀졌고 용매로 추출이 불가능한 Bentazon 토양 잔류물은 25.05 ㎎/㎏을 처리한 토양에서보다는 5.51 ㎎/㎏을 처리한 토양에서 모두 최초 시용량에 대하여 보다 높은 비율로 형성되었다.

• Journal of Pharmaceutical Investigation
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• 제34권2호
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• pp.101-106
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• 2004

This work aims at examining the cellular uptake behavior of poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles derivatized with a protein transduction domain (PTD) using HeLa cells. For this purpose, $Tat_{49-57}$ peptide derived from transcriptional activation (Tat) protein of HIV type-1 was covalently conjugated to the terminal end of PLGA. Nanoparticles were ten prepared with the $Tat_{49-57}-PLGA$ conjugates by a spontaneous phase inversion method. The prepared particles had a mean diameter of ca. 84 nm, as measured by dynamic light scattering. The interaction of the Tat-PLGA nanoparticles with cells was examined by using confocal laser scanning microscopy. It was found tat Tat-PLGA nanoparticles incubated with HeLa cells could efficiently translocate into cytoplasm, while plain PLGA nanoparticles showed negligible cellular uptake. In addition, even at $4^{\circ}C$ or in the presence of sodium azide significant cellular internalization of Tat-PLGA nanoparticles was still observed. These results indicate that a non-endocytotic translocation mechanism might be involved in the cellular uptake of Tat-PLGA nanoparticles.

• 한국수정란이식학회지
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• 제24권3호
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• pp.183-187
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• 2009

In this study, we aimed to determine whether the evaluated markers of cell death could be found at particular developmental stages of normal porcine in vitro fertilization (IVF) embryos. We investigated the characteristics of spontaneous and induced apoptosis during preimplantation development stages of porcine IVF embryos. In experiment 1, to induce apoptosis of porcine IVF embryos, porcine IVF embryos at 22h post insemination were treated at different concentration of actinomycin D (0, 5, 50 and 500 ng/ml in NCSU medium). Treated embryos were incubated at $39^{\circ}C$ in 5% $CO_2$, 5% $O_2$ for 8h, and then washed to NCSU medium and incubated until blastocyst (BL) stage. We examined cleavage rate at 2days and BL development rate at 7days after in vitro culture. A significantly lower rate of cleavage was found in the 500 ng/ml group compared to others (500 ng/ml vs. 0, 5, 50 ng/ml; 27.8 % vs. 50.0%, 41.2%, 35.9%), and BL formation rate in 500 ng/ml was lower than that of others (500 ng/ml vs. 0, 5, 50 ng/ml; 8.0% vs. 12.6%, 11.2%, 12.6%). In experiment 2, to evaluate apoptotic cells, we conducted TUNEL assay based on morphological assessment of nuclei and on detection of specific DNA degradation under fluorescence microscope. This result showed that apoptosis is a normal event during preimplantation development in control group (0 ng/ml actinomycin D). A high number of BL derived control group contained at least one apoptotic cell. Actinomycin D treated BLs responded to the presence of apoptotic inductor by significant decrease in the average number of blastomeres and increase in the incidence of apoptotic cell death. In 500 ng/ml group, the incidence of apoptosis increased at 4-cell stage and later. This result suggested that apoptosis is a process of normal embryonic development and actinomycin D is useful tool for the apoptosis study of porcine preimplantation embryos.

• Asian-Australasian Journal of Animal Sciences
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• 제21권3호
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• pp.358-363
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• 2008

The intracytoplasmic sperm injection (ICSI) procedure has recently been utilized to produce transgenic animals and may serve as an alternative to the conventional pronuclear microinjection in species such as pigs whose ooplasm is opaque and pronuclei are often invisible. In this study, the effects of sperm membrane disruption and electrical activation of oocytes on in vitro development and expression of transgene green fluorescent protein (GFP) in ICSI embryos were tested to refine this recently developed procedure. Prior to ICSI, sperm heads were treated with Triton X-100+NaCl or Triton X-100+NaCl+NaOH, to disrupt membrane to be permeable to exogenous DNA, and incubated with linearized pEGFP-N1 vector. To induce activation of oocytes, a single DC pulse of 1.3 kV/cm was applied to oocytes for $30{\mu}sec$. After ICSI was performed with the aid of a micromanipulator, in vitro development of embryos and GFP expression were monitored. The chemical treatment to disrupt sperm membrane did not affect the developmental competence of embryos. 40 to 60% of oocytes were cleaved after injection of sperm heads with disrupted membrane, whereas 48.6% (34/70) were cleaved without chemical treatment. Regardless of electrical stimulation to induce activation, oocytes were cleaved after ICSI, reflecting that, despite sperm membrane disruption, the perinuclear soluble sperm factor known to mediate oocyte activation remained intact. After development to the 4-cell stage, 11.8 (2/17, Triton X-100+NaCl+NaOH) to 58.8% (10/17, Triton X-100+NaCl) of embryos expressed GFP. The expression of GFP beyond the stage of embryonic genome activation (4-cell stage in the pig) indicates that the exogenous DNA might have been integrated into the porcine genome. When sperm heads were co-incubated with exogenous DNA following the treatment of Triton X-100+NaCl, GFP expression was observed in high percentage (58.8%) of embryos, suggesting that transgenic pigs may efficiently be produced using ICSI.

• Parasites, Hosts and Diseases
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• 제49권2호
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• pp.177-180
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• 2011

Entamoeba histolytica is an enteric tissue-invading protozoan parasite that can cause amebic colitis and liver abscess in humans. E. histolytica has the capability to kill colon epithelial cells in vitro; however, information regarding the role of calpain in colon cell death induced by ameba is limited. In this study, we investigated whether calpains are involved in the E. histolytica-induced cell death of HT-29 colonic epithelial cells. When HT-29 cells were co-incubated with E. histolytica, the propidium iodide stained dead cells markedly increased compared to that in HT-29 cells incubated with medium alone. This pro-death effect induced by ameba was effectively blocked by pretreatment of HT-29 cells with the calpain inhibitor, calpeptin. Moreover, knockdown of m- and ${\mu}$-calpain by siRNA significantly reduced E. histolytica-induced HT-29 cell death. These results suggest that m- and ${\mu}$-calpain may be involved in colon epithelial cell death induced by E. histolytica.

• 한국식품영양학회지
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• 제27권5호
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• pp.845-850
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• 2014

In the present study, the effects of extracts from Korean plants on the DNA damage response in HaCaT cells exposed to ultraviolet B (UVB) were investigated. The activity of cells treated for 24 hr with ethanol extracts from Vaccinium spp. (VS), and Vitis vinifera L (VV) alone was similar to that of the non-treated control, but gradually decreased at concentrations above $200{\mu}g/mL$. However, when post-incubation of UVB-exposed cells was carried out for 24 hr in medium containing VS or VV extracts, the cell activity increased in a concentration-dependent manner compared with that in the normal growth medium. The cell viability of UVB-exposed cells also increased when post-incubated in medium containing VS or VV extracts, in a concentration-dependent manner. Nuclear fragmentation analysis showed that post-incubation with VS or VV extracts decreased the UVB-induced apoptosis by about 10 and 13%, respectively, of that in cells post-incubated in growth medium. After 24 hr of post-incubation in medium containing VS or VV extracts, the level of CPD and 8-OHdG decreased in time- and concentration-dependent manners. Overall these results suggest that VS and VV extracts assist the survival of UVB-exposed cells, in accordance with the respective decrease in the levels of UVB-induced DNA damage.

• 한국초지조사료학회지
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• 제33권3호
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• pp.197-205
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• 2013

An in vitro study was conducted to determine the effects of defaunation (removal of protozoa) and forage sources (rice straw, ryegrass and tall fescue) on ruminal fermentation characteristics, methane ($CH_4$) production and degradation by rumen microbes. Sodium lauryl sulfate, as a defaunation reagent, was added into the mixed culture solution to remove ruminal protozoa at a concentration of 0.375 mg/ml. Pure cellulose (0.64 g, Sigma, C8002) and three forage sources were incubated in the bottle of culture solution of mixed rumen microbes (faunation) or defaunation for up to 24 h. The concentration of ammonia-N was high under condition of defaunation compared to that from faunation in all incubations (p<0.001). Total VFA concentration was increased at 3, 6 and 12 h (p<0.05~p<0.01) but was decreased at 24 h incubation (p<0.001) under condition of defaunation. Defaunation decreased acetate (p<0.001) and butyrate (p<0.001) proportions at 6, 12 and 24 h incubation times, but increased propionate (p<0.001) proportion at all incubation times for forages. Effective degradability of dry matter was decreased by defaunation (p<0.001). Defaunation not only decreased total gas (p<0.001) and $CO_2$ (p<0.01~0.001) production at 12 and 24 h incubations, but reduced $CH_4$ production (p<0.001) at all incubation times for all forages. The $CH_4$ production, regardless of defaunation, in order of forage sources were rice straw > tall fescue > ryegrass > cellulose (p<0.001) up to 24 h incubation.

• 대한치과의사협회지
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• 제46권10호
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• pp.623-632
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• 2008

Purpose : The purpose of this study is to identify the effect of zoledronate(Zometa(R)), which is most common nitrogen containing bisphosphonate, on survival, proliferation, and differentiation of osteoblast. Material & Methods: Twenty four cell culture plates containing essential medium were seeded with UMR-106 cell lines, at density of 5 x $10^4 cells per plates. Each plates were incubated with 5% $CO^2 incubator at $37^{\circ}C$. Starting from 2 days after incubation, cell culture medias were replaced, and added with osteogenesis induction media and 0, 0.01, 0.1, 0.5, 1, $3\muM$ of zoledronate(Zometa(R)), every 2 days, for 12 days. Control group was plates not added with zoledronate($0\muM$), and experiment group were plates added with different concentration of zoledronates(0, 0.01, 0.1, 0.5, 1, $3\muM$). Mature osteoblasts were identified with Alizarine Red staining, and protein samples were collected. Optical density was determined at wavelength of 405nm with ELISA reader. For viability analysis, cells were harvested and incubated with propidium iodide, and analysed with flow cytometry. Western blot technique was used to analyse Runx2 protein of osteoblast. Results : Secretion of bone matrix decreased as zoledronate concentration increased, and zoledronate did not effect survival rate of UMR-106 cells when measured with flow cytometer. Expression of Runx2 protein was inhibited as zoledronate concentration increased. Conclusion : From the results, we were able to identify that increase of zoledronate concentration inhibited differentiation of UMR-106 cell to osteoblast, without effecting quantity or survival rate.

• Journal of Neurogastroenterology and Motility
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• 제24권4호
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• pp.669-675
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• 2018

Background/Aims Functional dyspepsia (FD) and irritable bowel syndrome (IBS) are common gastrointestinal (GI) disorders and these patients frequently overlap. Trimebutine has been known to be effective in controlling FD co-existing diarrhea-dominant IBS, however its effect on overlap syndrome (OS) patients has not been reported. Therefore, we investigated the effect of trimebutine on the model of OS in guinea pigs. Methods Male guinea pigs were used to evaluate the effects of trimebutine in corticotropin-releasing factor (CRF) induced OS model. Different doses (3, 10, and 30 mg/kg) of trimebutine were administered orally and incubated for 1 hour. The next treatment of $10{\mu}g/kg$ of CRF was intraperitoneally injected and stabilized for 30 minutes. Subsequently, intragastric 3 mL charcoal mix was administered, incubated for 10 minutes and the upper GI transit analyzed. Colonic transits were assessed after the same order and concentrations of trimebutine and CRF treatment by fecal pellet output assay. Results Different concentrations (1, 3, and $10{\mu}g/kg$) of rat/human CRF peptides was tested to establish the OS model in guinea pigs. CRF $10{\mu}g/kg$ was the most effective dose in the experimental OS model of guinea pigs. Trimebutine (3, 10, and 30 mg/kg) treatment significantly reversed the upper and lower GI transit of CRF induced OS model. Trimebutine significantly increased upper GI transit while it reduced fecal pellet output in the CRF induced OS model. Conclusions Trimebutine has been demonstrated to be effective on both upper and lower GI motor function in peripheral CRF induced OS model. Therefore, trimebutine might be an effective drug for the treatment of OS between FD and IBS patients.

• Biomolecules & Therapeutics
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• 제30권2호
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• pp.170-178
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• 2022

The airway epithelium is equipped with the ability to resist respiratory disease development and airway damage, including the migration of airway epithelial cells and the activation of TLR3, which recognizes double-stranded (ds) RNA. Primary cilia on airway epithelial cells are involved in the cell cycle and cell differentiation and repair. In this study, we used Beas-2B human bronchial epithelial cells to investigate the effects of the TLR3 agonist polyinosinic:polycytidylic acid [Poly(I:C)] on airway cell migration and primary cilia (PC) formation. PC formation increased in cells incubated under serum deprivation. Migration was faster in Beas-2B cells pretreated with Poly(I:C) than in control cells, as judged by a wound healing assay, single-cell path tracking, and a Transwell migration assay. No changes in cell migration were observed when the cells were incubated in conditioned medium from Poly(I:C)-treated cells. PC formation was enhanced by Poly(I:C) treatment, but was reduced when the cells were exposed to the ciliogenesis inhibitor ciliobrevin A (CilioA). The inhibition of Beas-2B cell migration by CilioA was also assessed and a slight decrease in ciliogenesis was detected in SARS-CoV-2 spike protein (SP)-treated Beas-2B cells overexpressing ACE2 compared to control cells. Cell migration was decreased by SP but restored by Poly(I:C) treatment. Taken together, our results demonstrate that impaired migration by SP-treated cells can be attenuated by Poly(I:C) treatment, thus increasing airway cell migration through the regulation of ciliogenesis.