• Toxicological Research
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• 제13권3호
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• pp.251-256
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• 1997

Toluene inhalation increases glutamate level and its receptor in various brain regions. In this study, nitric oxide synthase (NOS) activities were investigated in various rat brain regions using NADPH diaphorase staining method which examined histochemical changes of NOS in the neural cells. Also, in vitro LDH leakage assay and MTT test were performed to investigate the toxic influences of toluene in cultured granule cell of rat cerebellum which was significantly affected with toluene in vivo. Rats were exposed to toluene of 10000 ppm for 3 days. 7 days and 14 days by 20 min $\times$ 2 times a day. NADPH diaphorase staining was processed in the different brain regions after inhalation. NADPH diaphorase staining density was not significantly changed at 3 days inhalation group, but the density decreased in proportion to the duration of toluene inhalation. Over 30% of staining density was decreased at 14 days group which was maximum duration of inhalation in this study. The tendency of staining density decrease was significant in granule cell of cerebellum. Cell death by toluene exposure was observed in cultured cerebellar granule cell. $EC_{50}$ measured with LDH leakage assay and MTT test were 43 mM and 72 mM respectively.

• 대한소아치과학회지
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• 제23권2호
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• pp.559-574
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• 1996

The purpose of this study was to determine whether there was any correlation between temporomandibular joint dysfunction and structure of the mandibular condyle. Weanling rats had their masseter muscles resected and immunohistochemical findings were observed with a light microscope. The results obtained were as follows : 1. The condylar cartilage region was divided into articular, proliferating, cartilage cell and hypertrophic cell layers according to cell morphology. 2. In light microscopic views, the proliferating and cartilage cell layers of the experimental group decreased gradually and at the 8th week significantly. 3. In immunohistochemical staining for type I and II collagen, a reaction was detected in the lower part of proliferating cell and cartilage cell layers. In the cartilage cell layers, a stronger cellular reaction was present. Immunohistochemical staining for type II collagen reacted more strongly than that of type I collagen. 4. In immunohistochemical staining for proteoglycan, the staining of the experimental group resembled the control group and gradually showed a weak reaction. The proliferating and cartilage cell layers reacted more strongly than the hypertrophic cell layer. 5. In immunohistochemical staining for proliferating cell nuclear antigen(PCNA), the strong reaction was detected in the nucleus of the proliferating cell layer both in control and experimental groups. But the thickness of the proliferating layer decreased in experimental group, consequently the reaction of the experimental group was reduced more than that of the control group.

• Environmental Engineering Research
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• 제23권3호
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• pp.282-287
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• 2018

Three different methods of bacterial viability monitoring were compared to detect chemically or thermally inactivated Escherichia coli. Direct colony enumeration, live/dead bacterial cell staining with a fluorescent dye, and the dehydrogenase activity assay were compared with respect to their ease of use and time required to perform the three different tests. The green (live cell)/red (dead cell) ratio obtained from the fluorescent bacterial cell staining approach showed a linear relationship with the colony forming units; the result obtained with dehydrogenase was similar to those. The sensitivity of the monitoring methods to detect bacterial deactivation varied with different disinfection conditions. After thermal treatment, the sensitivity of the staining approach was lower, while that of the dehydrogenase activity assay was the highest. After chemical treatment, the sensitivity of detection for both methods was similar.

• 대한수의학회지
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• 제35권2호
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• pp.317-326
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• 1995

The purpose of this study was to establish a method for in vitro culture of C parvum isolated in Korea by determination of suitable cell model to complete development of this parasite. The result obtained were summerized as follows: 1. To determine the most suitable cell line, six types of cell line were examined by microscopy. All cell lines were infected with C parvum and showed the highest infection score in HmLu cells. 2. The staining methods including DMSO-modified acid-fast(A-F) stain, hematoxylin-eosin(H & E) stain and immunofluorescence antibody(IFA) stain were applied to examine the infection of C parvum in cell culture. These staining methods were possible to examine the infection of C parvum in cell culture. The most sensitive one was IFA staining technique. 3. Developmental stages of C parvum in HmLu cell were observed. After the initial 8 hour incubation period, some trophozoites were observed. The meronts and gametes were appeared at 24-48 hour post inoculation(PI), and oocysts were observed firstly at 48-72 hour PI. 4. In H & E stain, the parasite appeared as basophilic within parasitophorous vacuole membrane(PVM) and lying in cytoplasm at near the nucleus of the host cells. It was able to distinguish the type I, type II meronts and gametes. 5. In DMSO-modified acid-fast stain, specific stained parasites were appeared firstly after 48 hour PI. The parasites were showed with different degrees of staining bright red color within PVM. 6. The endogenous stages of parasites in HmLu cell recovered at 48, 96, 120 and 144 hour after inoculation were reacted with rabbit immunized serum in immunofluorescence antibody and avidin-biotin complex peroxidase staining technique.

• 한국동물학회지
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• 제8권1호
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• pp.11-14
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• 1965

The object to this study is to identify the genus and species of the microfilariae which were recently found in the area of Youngju-Gun. THe identification of the microfilariae was made on the morphologicla aspects. 1. Blood samples were collected through vena punction from known microfilariae carriers living in the newly confirmed filaria endemic area of Youngju0Gun in Jyongsang Pukdo Province. Youngju-Gun is located in the mountainous central part of Korean Peninsula. 2. The following fixation and staining techniques were applied. (1) Fixation by drying in the air, followed by staining with Azeo or Giemsa. (2) Knott's fixation method (2% formalin), followed by staining with Azur II. 3. A comparative study of the body length of the microfilariae after different fixation and staining techniques were applied. (1) Knott's fixation method followed by staining with Azur II : average body length found was 28.4$\mu$. (2) Dry fixation followed by staining with Giemsa : average body length found was 209.4$\mu$. (3) Dry fixation followed by staining with Azeo : average body length found was 205.4$\mu$. 4. The locations of the different body cells were measured in 60 individuals of microfilariae in the wet preparation fixed by Knott's method and stained with Azur II. The distance of the different body cells to cephalic apex of microfilariae was measured and calculated as a percentage of the total body length. The average results are as follows : BNC , 3.04% ; N, 22.74%, EP, 31.4% ; EC, 37.77%, G1 cell , 67.94%; G2 cell, 73.54% : G3 cell , 75.55% ; G4 cell, 77.65% ; AP, 82.02%%. 5. As a result of the above findings the microfilariae found in the above mentioned area could be identified as Brugia malai(BRUG, 1927) BUCKLEY, 1960.

• 생약학회지
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• 제28권4호
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• pp.225-232
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• 1997

300 extracts derived from 100 plants were tested for their potential to induce HL-60 cell differentiation using NBT assay and NSE/SE staining methods. Morphological changes from suspended to adherent state of the cells were also observed by microscopic examination. In result, 55 extracts induced cell differentiation into monocyte/macrophage lineage in the NBT and the NSE assay.

• Tuberculosis and Respiratory Diseases
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• 제48권3호
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• pp.324-330
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• 2000

연구배경 : IGFs는 다양한 종양세포에서 세포분열 및 성장에 관여하는 것으로 알려진 펩티드로써 폐암 조직에서 IGF-1에 대한 항체를 이용하여 면역조직화학염색을 실시하여 폐암세포에서 이의 발현 및 조직학적 형태에 따라 발현의 정도를 비교해 보고자 하였다. 방 법 : 15명의 소세포성 폐암 환자와 42명의 비소세포성 폐암 환자를 대상으로 IGF-1에 대한 면역조직화학적 염색을 실시하였다. 결 과 : 모든 폐암 조직애서 IGF-1의 발현을 보였고 비소세포성 폐암조직은 소세포성 폐암조직보다 IGF-1에 대한 발현의 정도가 유의하게 증가되어 있었다. 결 론 : 폐암세포는 IGF-1의 발현을 보이며 이에 대한 면역조직화학염색은 폐암세포의 조직학적 형태를 감별하는데 도움을 줄 수 있다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권5호
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• pp.2885-2889
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• 2013

Goniothalamin is an active compound extracted from Goniothalamus griffithii, a local plant found in northern Thailand. Goniothalamin inhibits cancer cell growth but is also toxic to normal cells. The aims of this study were to identify the cytotoxic effect of goniothalamin and the mechanism of cell death in human HL-60 and U937 cells. Cytotoxicity was determined by MTT assay and cell cycle profiles were demonstrated by staining with propidium iodide (PI) and flow cytometry. Apoptosis was confirmed by staining with annexin V-FITC/propidium iodide (PI) and flow cytometry. Reduction of mitochondrial transmembrane potential was determined by staining with dihexyloxacarbocyanine iodide and flow cytometry and expression of Smac, caspase-8 and -9 was demonstrated by Western blotting. Goniothalamin inhibited growth of HL-60 and U937 cell lines. An increase of SubG1 phase was found in their cell cycle profiles, indicating apoptosis as the mode of cell death. Apoptosis was confirmed by the flip-flop of phosphatidylserine using annexin V-FITC/PI assay in HL60 and U937 cells in a dose response manner. Furthermore, reduction of mitochondrial transmembrane potential was found in both cell types while expression of caspase-8, -9 and Smac/Diablo was increased in HL-60 cells. Taken together, our results indicate that goniothalamin-treated human leukemic cells undergo apoptosis via intrinsic and extrinsic pathways.

• Applied Microscopy
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• 제42권4호
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• pp.200-206
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• 2012

Induction of neurogenesis can occur in the hippocampus in response to various pathological conditions, such as Alzheimer's disease. The aim of this study was to investigate the changes that occur in endogenous neural stem cells in response to amyloid beta $(A{\beta})_{25-35}$-induced neuronal cell damage in organotypic hippocampal slice cultures. Cresyl violet staining and Fluoro-Jade B staining were used to detect neuronal cell damage and changes of mossy fiber terminals were observed by Timm's staining. The immunofl uorescence staining was used to detect the newly generated cells in the subgranular zone (SGZ) of the dentate gyrus with specific marker, 5-bromo-2'-deoxyuridine (BrdU), Ki-67, Nestin, and doublecortin (DCX). In compared to control slices, neuronal cell damage was observed and the mossy fibers were expanded to CA3 area by treatment with $A{\beta}_{25-35}$. Ki-67/Nestin- and BrdU/DCX-positive cells were detected in the SGZ. In conclusion, these results demonstrate that $A{\beta}$-induced neuronal damage results in an increase in endogenous neural stem cells in rat hippocampal slice cultures not only for gliosis but also for neurogenesis.

• Journal of Ginseng Research
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• 제47권4호
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• pp.572-582
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• 2023

Background: Free fatty acid-induced lipotoxicity is considered to play an important role in pancreatic β-cell dysfunction. The effect of ginsenosides on palmitic acid-induced pancreatic beta-cells cell death and failure of glucose-stimulated secretion of insulin (GSIS) was evaluated in this study. Methods: Enzyme-linked immunosorbent assay kit for a rat insulin was used to quantify glucose-stimulated insulin secretion. Protein expression was examined by western blotting analysis. Nuclear condensation was measured by staining with Hoechst 33342 stain. Apoptotic cell death was assessed by staining with Annexin V. Oil Red O staining was used to measure lipid accumulation. Results: We screened ginsenosides to prevent palmitic acid-induced cell death and impairment of GSIS in INS-1 pancreatic β-cells and identified protopanaxadiol (PPD) as a potential therapeutic agent. The protection effect of PPD was likely due to a reduction in apoptosis and lipid accumulation. PPD attenuated the palmitic acid-induced increase in the levels of B-cell lymphoma-2-associated X/B-cell lymphoma 2, poly (ADP-ribose) polymerase and cleaved caspase-3. Moreover, PPD prevented palmitic acid-induced impairment of insulin secretion, which was accompanied by an increase in the activation of phosphatidylinositol 3-kinase, peroxisome proliferator-activated receptor γ, insulin receptor substrate-2, serine-threonine kinase, and pancreatic and duodenal homeobox-1. Conclusion: Our results suggest that the protective effect of PPD on lipotoxicity and lipid accumulation induced by palmitic acid in pancreatic β-cells.