• Journal of the Korean Wood Science and Technology
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• v.41 no.3
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• pp.247-257
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• 2013

This study was to investigate the antifungal activity of Larix kaempferi essential oil against dermatophytes, Epidermophyton floccosum, Trichophyton mentagrophytes and Trichophyton rubrum. The active components of L. kaempferi against dermatophytes were determined (characterized by GC-MS), and the morphological changes of the dermatophytes exposed to the L. kaempferi essential oil were observed by electron microscope. Main component of L. kaempferi essential oil was (-)-bornyl acetate. In antifungal activity tests, MIC of L. kaempferi crude oil was 125 ppm on every fungi and 100% (agar dilution method) at more than 500 ppm. By using SEM and TEM, the fungal morphology of E. floccosum exposed to the L. kaempferi essential oil was different from that of normal hyphal morphology. Hyphae exposed to the L. kaempferi essential oil was damaged with distorted and collapsed surfaces. In addition, there were destruction and disorganization of organelles in cytoplasm and collapse of cell membrane. Active antifungal components from L. kaempferi essential oil were identified as terpene alcohol compounds like (-)-${\tau}$-muurolol, (+)-terpinen-4-ol, ${\alpha}$-terpineol, and ${\alpha}$-cadinol.

• Molecules and Cells
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• v.41 no.3
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• pp.224-233
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• 2018

Primary cilia are solitary, non-motile, axonemal microtubule-based antenna-like organelles that project from the plasma membrane of most mammalian cells and are implicated in transducing hedgehog signals during development. It was recently proposed that aberrant SHH signaling may be implicated in the progression of idiopathic pulmonary fibrosis (IPF). However, the distribution and role of primary cilia in IPF remains unclear. Here, we clearly observed the primary cilia in alveolar epithelial cells, fibroblasts, and endothelial cells of human normal lung tissue. Then, we investigated the distribution of primary cilia in human IPF tissue samples using immunofluorescence. Tissues from six IPF cases showed an increase in the number of primary cilia in alveolar cells and fibroblasts. In addition, we observed an increase in ciliogenesis related genes such as IFT20 and IFT88 in IPF. Since major components of the SHH signaling pathway are known to be localized in primary cilia, we quantified the mRNA expression of the SHH signaling components using qRT-PCR in both IPF and control lung. mRNA levels of SHH, the coreceptor SMO, and the transcription factors GLI1 and GLI2 were upregulated in IPF compared with control. Furthermore, the nuclear localization of GLI1 was observed mainly in alveolar epithelia and fibroblasts. In addition, we showed that defective KIF3A-mediated ciliary loss in human type II alveolar epithelial cell lines leads to disruption of SHH signaling. These results indicate that a significant increase in the number of primary cilia in IPF contributes to the upregulation of SHH signals.

• Korean Journal of Environmental Biology
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• v.25 no.4
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• pp.289-296
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• 2007

Di-(ethyhexyl) phthalate (DEHP), commonly used as a plasticizer for manufacturing flexible vinyl products, has been the topic of extensive research, especially concerning endocrine disrupting properties. Metallothionein (MT) is a low molecular weight (6,000$\sim$7,000 Da), cysteine-rich (22$\sim$23%), metal-binding protein and is known to be induced by extrinsic factors such as chemical agents and stresses. Some of the known function of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Nonetheless, the definitive physiological function of MT are still unknown. This study was carried out to investigate the effects of DEHP on the ultrastructural changes and the expression of MT of the rat liver. The rats were orally intubated with either corn oil (experimental control) or 0.5 mg, 1.5 mg and 4.5 mg DEHP kg$^{-1}$ day$^{-1}$ in 0.5 mL of corn oil for 15 days before sacrificing and sampling. DEHP induced mild ultrastrctural changes of some cell organelles such as rough endoplasmic reticulum, mitochondria, lysosomes and peroxisomes in the rat liver treated with DEHP. In the respect of immunogold labelling and Western blotting, MT expression of the liver tissue was up-regulated by DEHP. In conclusion, DEHP has effects on the ultrastructures and hepatic function for MT expression in rat.

• Applied Microscopy
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• v.33 no.1
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• pp.79-92
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• 2003

Ultrastructural changes of the pericarp in Citrus reticulata has been investigated during hesperidium abscission. The pericarp was composed of compactly arranged parenchyma cell layers during early stages of fruit development. The outermost exocarp was green and active in photosynthesis. However, cells in the exocarp soon changed into collenchyma cells by developing unevenly thickened walls within a short time frame. As the fruit approached maturation, the chlorophyll gradually disappeared and chloroplasts were transformed into carotenoid-rich chromoplasts. In the mature fruit the exocarp consisted of large, lobed collenchyma cells with primary pit fields and numerous plasmodesmata. The immature mesocarp was a relatively hard and thick layer, located directly under the exocarp. With development, the deeper layers of the exocarp merged into the white, spongy mesocarp. Before separation of the hesperidium from the plant, some unusual features were detected in the plasma membrane of the exocarp cells. The number of small vacuoles and dark, irregular osmiophilic lipid bodies also increased enormously in the exocarp collenchyma after the abscission. They occurred between the plasma membrane and the wall, and invaginated pockets of the plasma membrane containing double-membraned vesicles were also frequently noticed. The lipid bodies in the cytoplasm were often associated with other organelles, especially with plastids and mitochondria. The plastids, which were irregular or amoeboid in shape, contained numerous large lipid droplets, and occasional clusters of phytoferritin, as well as few loosely -oriented peripheral lamellae. Myelin-like configurations of membrane were frequently observed in the vacuoles, as was the association of lipid bodies with the vacuolar membrane. Most vacuoles had an irregular outline, and lipid bodies were often connected to the tonoplast of the vacuoles. The structural changes underlying developmental, particularly to senescence, processes in various hesperidium will be reported in the separate paper.

• Journal of Applied Biological Chemistry
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• v.58 no.3
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• pp.219-226
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• 2015

The anticancer activity of a methanolic extract from lemon leaves (MLL) was assessed in MCF-7-SC human breast cancer stem cells. MLL induced apoptosis in MCF-7-SC, as evidenced by increased apoptotic body formation, sub-G1 cell population, annexin V-positive cells, Bax/Bcl-2 ratio, as well as proteolytic activation of caspase-9 and caspase-3, and degradation of poly (ADP-ribose) polymerase (PARP) protein. Concomitantly, MLL induced the formation of acidic vesicular organelles, increased LC3-II accumulation, and reduced the activation of Akt, mTOR, and p70S6K, suggesting that MLL initiates an autophagic progression in MCF-7-SC via the Akt/mTOR pathway. Epithelial-mesenchymal transition (EMT), a critical step in the acquisition of the metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. At low concentrations, MLL induced anti-metastatic effects on MCF-7-SC by inhibiting the EMT process. Exposure to MLL also led to an increase in the epithelial marker E-cadherin, but decreased protein levels of the mesenchymal markers Snail and Slug. Collectively, this study provides evidence that lemon leaves possess cytotoxicity and anti-metastatic properties. Therefore, MLL may prove to be beneficial as a medicinal plant for alternative novel anticancer drugs and nutraceutical products.

• Applied Microscopy
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• v.36 no.4
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• pp.279-289
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• 2006

This study has been carried out to understand the effect of SP3 and GV6 acupuncture on the hyperglycemic rat induced with Streptozotocin (STZ). Diabetes was induced in experimental groups by inraperitoneal injection of STZ (50mg/kg of body weight) twice by 24 h interval and the additional 100mg/kg 3 days after the earlier treatment. Control group was treated with tail-non acupoint, and experimental groups were treated SP3, GV6 and SP3+GV6 after hyperglycemic induction for 6 weeks. The body weight of control was lower than the experimental groups. The blood glucose was decreased significantly in the experimental groups. Glucose tolerance in acupuncture treated groups was improved. Blood cholesterol level and transaminase activites were lower in the experimental groups than in the control. In the SP3 or GV6 treated rats, hepatocyte degeneration were apparently decreased and the organelles were properly arranged. Furthermore, decrease in liver IGF-I mRNA expression was improved by the acupuncture in STZ-induced diabetic rats. In conclusion, our observation indicate that SP3 or GV6 acupoints treatment can exert beneficial effects in diabetes, with preservation of ${\beta}-cell$ function and liver function.

• Applied Microscopy
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• v.29 no.1
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• pp.57-66
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• 1999

The nervous tissue in the cerebral ganglion of Korean planaria was observed using electron microscope. The obtained results are as follows: A cerebral ganglion is composed of the nerve cells, neurosecretory cells, neuroglial cells and neuropils. The nerve cells are round or ovoidal-shaped cells (diameter, $5{\mu}m$), which has a large ellipsoidal nucleus containing the evenly developed heterochromatin. Their cytoplasms were found to be relatively simple, because of their undeveloped cell organelles. The neurosecretory cells are long and ellipsoid or spindle-shaped cells, where there were found a large ellipsoidal nucleus and cytoplasm filled with secretory granules (diameter, 60 nm). The neuroglial cells were seldom observed. They are spindle-shaped cells (size, $6\times0.8{\mu}m$), which were observed mainly among the nerve fibers. The neuropils are formed by the nerve fibers and nerve endings which are filled with mitochondria, neurotubules and secretory granules of four kinds (high electron dense granules of sizes 75 nm, 50 nm and 37 nm, and electron lucent granule of size 30 nm etc.). These granular vesicles are divided into single vesicle type and compound vesicle type in the nerve terminals, and neuronal synapses were observed to be the axo-dendritic and dendro-dendritic synapse type.

• Parasites, Hosts and Diseases
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• v.36 no.1
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• pp.7-14
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• 1998

In order to understand the action mechanism of polyhexamethylene biguanide (PHMB) to the cyst of Accnthcnloebc on the morphological basis, the cysts of four corneal isolates of Acanthanoebc were treated with minimal cysticidal concentration (MCC) of PHMB and their ultrastructural changes were examined by transmission electron microscopy. The most striking change of cysts treated with PHMB compared with normal cysts was the shrinkage of intracystic amoebae, which resulted in the separation of the plasma membrane of intracystic amoeba from endocystic wall. Subplasmalemmal lipid droplets became irregularly shaped . In severely damaged cysts, cytoplasm was aggregated and organelles were severely deformed. Cytoplasmic materials were leaked out through the damaged plasma membrane. Most cysts showed aggregation of nuclear chromatin material. Number of mitochondrial cristae was also reduced. Ecto- and endo-cystic walls were relatively well tolerated. Findings in the present study revealed that PHMB affected mainly on plasma membrane, but lesser on organellar membrane of intracystic amoeba. It seemed likely that PHMB might kill cystic forms of Accnthamoebc by similar mechanism in which this environmental biocide can damage the cell wall of Escherichia coli by binding with acidic phospholipids.

• The Korean Journal of Pesticide Science
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• v.6 no.2
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• pp.96-104
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• 2002

This study was carried out to investigate cytotoxicity of paraquat on NIH 3T3 fibroblasts, toxicity of paraquat and compensatory effects of 3-methylcholanthrene (3-MC) on the rat lung. In order to conduct MIT [3-(4,5-Dimethylthiazol-2-yl) -2,5-diphenyl -2H-tetrazolium-bromide] and NR (Neutral red) assay, the $5.0{\times}10^4cell/ml$ of NIH 3T3 fibroblast in each well of 24 multi-dish were cultured. After 24 hours, the cells were treated with solution of paraquat (1, 25, 50 and $100{\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MIT and NR assay were performed to evaluate the cytotoxicity of cell organelles. $MTT_{50}\;and\;NR_{50}$ of paraquat were $1668.97{\mu}M\;and\;1030.85{\mu}M$, respectively. These $IC_{50}$ of Paraquat were decided as a low cytotoxicity by Borenfreund and Puemer (1984). In order to observe the toxicity and compensatory effects of paraquat on the rat lung, Spraque Dawley male rats were used as experimental animals and were divided into paraquat only treated group and simultaneous application group of paraquat and 3-MC, at 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment. The animals were sacrificed by decapitation and their or the lungs were immediately removed, immersed in fixatives, and were processed with routine method for light microscopic study. Paraffin sections were stained with H&E and iron hematoxylin of Verhoeff. Under the light microscopy, erythrocytes were full in alveolar capillaries at 3 hrs and congested at 24 hrs after paraquat administration. The great alveolar cells (Type II cell) were increased and mitosis of great alveolar were observed in interalveolar septa. Many lymphocytes, macrophages and polymorphonuclear (PMN) cells were observed in connective tissue surrounding lung tissue and germinal center in lymph follicles of terminal bronchiole. Alveolar macrophages were increased in interalveolar septa and alveoli at 48 hrs. And observed many alveolar macrophages at 96 hrs. In iron hematoxylin stain of Verhoeff, Collagen fiber were increased in respiratory bronchiole, interalveolar septa and alveoli and breath of alveoli, and alveolar pore were broaden. But, in paraquat plus 3-MC treated group, morphological changes were mild in lung tissue. These results indicate that 3-MC has a compensatory effects against toxicity of paraquat by conjugation with oxygen.

• Tuberculosis and Respiratory Diseases
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• v.69 no.1
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• pp.16-23
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• 2010

Background: Most lung cancer patients receive systemic chemotherapy at an advanced stage disease. Cisplatin-based chemotherapy is the main regimen for treating advanced lung cancer. Recently, autophagy has become an important mechanism of cellular adaptation under starvation or cell oxidative stress. The purpose of this study was to determine whether or not autophagy can occurred in cisplatin-treated lung cancer cells. Methods: H460 cells were incubated with RPMI 1640 and treated in $5{\mu}M$ or $20{\mu}M$ cisplatin concentrations at specific time intervals. Cells surviving cisplatin treatment were measured and compared using an MTT cell viability assay to cells that underwent apoptosis with autophagy by nuclear staining, apoptotic or autophagic related proteins, and autophagic vacuoles. The development of acidic vascular organelles was using acridine orange staining and fluorescent expression of GFP-LC3 protein in its transfected cells was observed to evaluate autophagy. Results: Lung cancer cells treated with $5{\mu}M$ cisplatin-treated were less sensitive to cell death than $20{\mu}M$ cisplatin-treated cells in a time-dependent manner. Nuclear fragmentation at $5{\mu}M$ was not detected, even though it was discovered at $20{\mu}M$. Poly (ADP-ribose) polymerase cleavages were not detected in $5{\mu}M$ within 24 hours. Massive vacuolization in the cytoplasm of $5{\mu}M$ treated cells were observed. Acridine orange stain-positive cells was increased according in time-dependence manner. The autophagosome-incorporated LC3 II protein expression was increased in $5{\mu}M$ treated cells, but was not detected in $20{\mu}M$ treated cells. The expression of GFP-LC3 were increased in $5{\mu}M$ treated cells in a time-dependent manner. Conclusion: The induction of autophagy occurred in $5{\mu}M$ dose of cisplatin-treated lung cancer cells.