• Applied Microscopy
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• v.36 no.3
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• pp.173-182
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• 2006

Recently, the pomegranate seed oil (PSO) has been reported to have various efforts including anti-cancer effect. In this study, we examined the liver-protecting effect of the PSO on the hepatotoxicity induced by $CCl_4$ using the BALB/c mice. The PSO was made from dried seeds of black pomegranate (Punica grantum) by heating and squeezing. The expreimental animals were divided into 3 groups; control group treated with olive oil only, experimental group 1 treated with $CCl_4$ only, and experimental group 2 treated with PSO and $CCl_4$. 24 hours after injection of $CCl_4$ into the peritoneal cavity, we collected the blood samples to measure the level of serological factors; aspartate aminotransferase(AST), alanine aminotransferase (ALT), total protein, albumin, total bilirubin, direct bilirubin and alkaline phosphatase. Simultaneously we observed the histological change of liver under the light and electron microscope. As the result, AST and ALT showed $88.7{\pm}14.9IU/L\;and\;22.0{\pm}3.12IU/L$ in the control group, $1963.7{\pm}1212.9IU/L\;and\;4495.4{\pm}2803.6IU/L$ in the experimental group 1, and $432.2{\pm}260.1IU/L\;and\;692.3{\pm}433.1IU/L$ in the experimental group 2. The experimental group 2 showed significant difference as compared with experimental group 1 (P<0.005). In histological study, the experimental group 2 was recovered than experimental group 1 which had abnormal mitochondria, increase of lysosomes, and severe necrosis at the central vein zones. These results indicated that the PSO had the liver protecting effect. However, The further study on the relationship between ingredients of pomegranate seed and liver protecting effect is in need.

• Applied Microscopy
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• v.41 no.4
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• pp.277-284
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• 2011

This study was conducted to determine the antiseptic effect of stabilized chlorine dioxide (S-$ClO_2$) on muscle tissue of rats. Skeletal muscle of 8-week old Sprague-Dawley rats was used. Light and transmission electron microscopic findings were observed in the control group, which was not treated with stabilized chlorine dioxide, and in the experimental group, which was treated with a stabilized chlorine dioxide powder in aqueous solution. According to the LM and TEM observations, the day 1 control group showed the initiation of endomysium collapse resulting in an unclear boundary of muscle fibers, and partial collapse of the mitochondrial membranes. All endomysium had collapsed, and bacteria were observed among muscle fibers in the day 2 and later groups. Shapes of muscles were not distinguishable in day 3 or later groups. In contrast, the day 1 and 3 experimental groups revealed detailed structure of typical muscles, but partial collapse of the mitochondrial membranes was observed in the day 3 and later groups. Subsequently, connective tissues collapsed and structures in the shape of concentric circles were observed. In summary, the day 1 control group showed the initial collapse of tissues, and shapes were not distinguishable in the day 3 and later groups because most of the tissues had collapsed. In contrast, the day 3 experimental group showed partial collapse, but the overall shapes of muscles were maintained as time went on, confirming the antiseptic effect of stabilized chlorine dioxide on muscles.

• Tuberculosis and Respiratory Diseases
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• v.58 no.1
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• pp.43-53
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• 2005

Emphysema is characterized by air space enlargement and alveolar destruction. The mechanism responsible for the development of emphysema was thought to be protease/antiprotease imbalance and oxidative stress. A very recent study shows that alveolar cell apoptosis causes lung destruction and emphysematous changes. Thus, this study was performed to support the evidence for the role of apoptosis in the development of emphysema by characterizing cigarette smoke extract (CSE)-induced apoptosis in A549 (type II pneumocyte) lung epithelial cells. CSE induced apoptosis at low concentration (10% or less) and both apoptosis and necrosis at high concentration (20%). Apoptosis was demonstrated by DNA fragmentation using FACScan for subG1 fraction. Discrimination between apoptosis and necrosis was done by morphologic analysis using fluorescent microscopy with Hoecst 33342/propium iodide double staing and electron microscopy. Cytochrome c release was confirmed by using immunofluorescence with monoclonal anti-cytochrome c antibody. However, CSE-induced cell death did not show the activation of caspase 3 and was not blocked by caspase inhibitors. This suggests that CSE-induced apoptosis might be caspase-independent apoptosis. CSE-induced cell death was near completely blocked by N-acetylcystein and bcl-2 overexpression protected CSE-induced cell death. This results suggests that CSE might induce apoptosis through intracellular oxidative stress. CSE also activated p53 and functional knock-out of p53 using stable overexpression of HPV-E6 protein inhibited CSE-induced cell death. The characterization of CSE-induced cell death in lung epithelial cells could support the role of lung cell apoptosis in the pathogenesis of emphysema.

• Korean Journal of Plant Taxonomy
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• v.34 no.2
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• pp.97-118
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• 2004

A comparative anatomical and ultrastructural study was undertaken to investigare on the leaf epidermis by light microscopy (LM) and scanning electron microscopy (SEM). On the basis of results from this study, it was grasped major characters of taxa and variation range of each character on the level of species, section and subgenus respectively. The shape of leaf epidermal cell was oblong to linear, which was varied by each taxon. Epidermal cell of taxa in sects. Microscordum, Anguinum, and Rhizirideum, which had wide leaf blade, oblong instead of linear shape in others examined taxa in this study. The leaf of taxa in sect. Anguinum was hypostomatic, while the rest of taxa had amphistomatic leaf. This was also one of characters which could discriminate taxa of sect. Anguinum from others. The guard cell in investigated taxa had not so much variation in the respect of its size. The number of stomata per unit area reduced by increasing size of epidermal cell, the fewest number of stomata per unit area was found in the taxa of sect. Anguinum. The type of stomatal apparatus of observed all taxa was anomocytic. It was found to know ultrastructural variation in the epidermal cell, like as patterns of sculpture on the cell wall, and features of deposition of wax by SEM. There were no depositions of wax in the taxa of sect. Microscordum and Anguinum, but fine thread-like structures which were parallel or cross to axis was found on the surface of epidermal cell respectively. The patterns of sculpture on the cell were prominent straight in sects. Recticulato-bulbosa and Rhizirideum, discontinuous line in the sect. Oreiprason. The epicuticular wax had been deposited on the surface of its epidermal cell in all taxa except sects. Microscordum and Anguinum.

• Journal of Korean Society of Environmental Engineers
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• v.34 no.5
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• pp.345-350
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• 2012

To investigate the exposure and health risk assessment for the residents near the D-asbestos mine in Chungbuk, Korea. We analyzed asbestos in the 20 ambient air and 23 activity based samples near the mine. The airborne sample results are showed that 8 of 20 samples ranged between 0.0025 to 0.0029 f/cc (fiber per cubic centimeter) and the others were below the detection limit by phase contrast microscopy (PCM). In addition, asbestos fibers were under the detection limit or not being by transmission electron microscopy (TEM). Based on interview and survey targeting the local residents, we made the activity based sampling (ABS) scenarios fit to the conditions of field. At the same time, we calculated the excess lifetime cancer risk (ELCR) of these ABS scenarios according to the ELCR average value and 95% upper confidence limit (UCL). At the case of weed whacking, soil digging and sweeping yard scenario, 95% UCL of ELCR exceeded the $1{\times}10^{-4}$, acceptable risk range for exposure. Based on our study results, it is necessary safety measures such as risk communication, abatement or management of naturally occurring asbestos (NOA).

• Journal of the Society of Cosmetic Scientists of Korea
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• v.29 no.2 s.43
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• pp.205-232
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• 2003

Ursolic acid (UA) and Oleanolic acid (ONA), known as urson, micromerol and malol, are pentacyclic triterpenoid compounds which naturally occur in a large number of vegetarian foods, medicinal herbs, and plants. They may occur in their free acid form or as aglycones for triterpenoid saponins, which are comprised of a triterpenoid aglycone, linked to one or more sugar moieties. Therefore UA and ONA are similar in pharmacological activity. Lately scientific research, which led to the identification of UA and ONA, revealed that several pharmacological effects, such as antitumor, hepato-protective, anti-inflammatory, anticarcinogenic, antimicrobial, and anti-hyperlipidemic could be attributed to UA and ONA. Here, we introduced the effect of UA and ONA on acutely barrier disrupted and normal hairless mouse skin. To evaluate the effects of UA and ONA on epidermal permeability barrier recovery, both flanks of 8-12 week-old hairless mice were topically treated with either 0.01-0.1 mg/ml UA or 0.1-1 mg/ml ONA after tape stripping, and TEWL (Transepidermal water loss) was measured . The recovery rate increased in those UA or ONA treated groups (0.1 mg/ml UA and 0.5 mg/ml ONA) at 6 h more than $20\%$ compared to vehicle treated group (p<0.05). Here, we introduced the effects of UA and ONA on acute barrier disruption and normal epidermal permeability barrier function. For verifying the effects of UA and ONA on normal epidermal barrier, hydration and TEWL were measured for 1 and 3 weeks after UA and ONA applications (2mg/ml per day). We also investigated the features of epidermis and dermis using electron microscopy (EM) and light microscopy (LM). Both samples increased hydration compared to vehicle group from f week without TEWL alteration (p<0.005). EM examination using RuO4 and OsO4 fixation revealed that secretion and numbers of lamellar bodies and complete formation of lipid bilayers were most prominent $(ONA{\geq}UA>Vehicle)$. LM finding showed that thickness of stratum corneum (SC) was slightly increased and especially epidermal thickening and flattening was observed (UA>ONA>Veh). We also observed that UA and ONA stimulate epidermal keratinocyte differentiation via $PPAR\;\alpha$. Protein expression of involucrin, loricrin, and filaggrin increased at least 2 and 3 fold in HaCaT cells treated with either $ONA\;(10{\mu}M)$ or UA $(10{\mu}M)$ for 24h respectively. This result suggested that the UA and ONA can improve epidermal permeability barrier function and induce the epidermal keratinocyte differentiation via $PPAR\;{\alpha}$. Using Masson-trichrome and elastic fiber staining, we observed collagen thickening and elastic fiber elongation by UA and ONA treatments. In vitro results of collagen and elastin synthesis and elastase inhibitory activity measurements were also confirmed in vivo findings. These data suggested that the effects of UA and ONA related to not only epidermal permeability barrier functions but also dermal collagen and elastic fiber synthesis. Taken together, UA and ONA can be relevant candidates to improve epidermal and dermal functions and pertinent agents for cosmeseutical applications.

• Korean Journal of Plant Taxonomy
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• v.33 no.4
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• pp.421-433
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• 2003

To examine the leaf epidermal microstructure, nine species in five genera (Daphne L. - 4 spp., Diarthron Turcz. - 1 sp., Edgewarthia Meisn. - 1 sp., Stellera L. - 1 sp., Wikstroemia Endl. - 2 spp.) of the Korean Thymelaeaceae were investigated by light microscopy (LM) and scanning electron microscopy (SEM). The stomata of stuo야ed taxa were 'hypostomatic type' and the size range of guard cell was $13.8-34.4{\times}8.7-22.9{\mu}m$: the smallest size of stomata was found in Diathran linifolium ($15.9{\pm}2.6{\times}10.0{\pm}1.3{\mu}m$), while the largest one was measured to Daphne adara ($32.8{\pm}1.6{\times}20.7{\pm}1.3{\mu}m$). The stomatal complex was anomocytic in the most studied taxa, except Daphne kiusiana by having combined with anisocytic together. The shapes of epidermal cells are undulate anticlinal wall. The size range of epidermal cell was $20.7-61.0{\mu}m$; the smallest size of epidermal cell was found in Stellera charnaejasme ($26.0{\pm}1.9{\mu}m$), on the other hand the largest one was found in Edgeworthia chrysantha ($53.6{\pm}3.1{\mu}m$). The well-developed flaky epicuticular waxes can be divided three kinds of pattern - (1) smooth in comparison, not entire platelets and scattered, (2) isolated flake-like platelets, mostly paralleled, sparsely, (3) flake-like platelets, flat, membraneous, protruding from the surfaces at varying angles and densely. Two types of trichome are recognized; (1) Type I: uniseriate trichome of striate surface (D. genkwa, Diarthron linifalium, E. chrysantha, W. ganpi and W. trichotama), (2) Type II: multicellular trichome of papillose surface, uncinated 3-4 nodes (Diathron linifolium). Finally, the systematics significance of the leaf micromorphological features in identification and elucidation of Korean Thymelaeaceae, especially between or within the genera including among the species is also briefly discussed.

• Restorative Dentistry and Endodontics
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• v.36 no.4
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• pp.290-299
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• 2011

Objectives: The usage of fluoride varnish for a moderate to low caries-risk group has not been well validated. This study aimed to evaluate the preventive and therapeutic efficacies of fluoride varnish on the initiated root caries. Materials and Methods: Ten premolars were sectioned into quarters, further divided into two windows, one of which was painted with Fluor Protector (1,000 ppm fluoride, Ivoclar Vivadent). An initial lesion with a well-preserved surface layer was produced by pH cycling. Scanned line analysis using energy dispersive spectrometry determined the weight percentages of Ca and P in the demineralized layer. Scanning Electron microscopy and confocal laser scanning microscopy (CLSM) evaluated the varnish-applied root surfaces. Results: The mean lesion depth (SD) was 12.3 (2.6) ${\mu}m$ (single cycling) and 19.6 (3.8) ${\mu}m$ (double cycling). Double cycling extended the lesion depth, but induced no more mineral loss than single cycling (p < 0.05). The mean weight percentages of Ca and P between groups with and without varnish were not significantly different (p < 0.05). A CLSM showed varnish remained within 15 ${\mu}m$ of the surface layer. Conclusions: When a mild acid challenge initiated root tissue demineralization, the application of low-concentration fluoride varnish did not influence the lesion depth or the mineral composition of the subsurface lesion.

• Applied Microscopy
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• v.37 no.1
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• pp.43-52
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• 2007

In order to observe the localization of excretory, purified and infected antigenic protein in the tissue of Trichinella spiralis larvae, immunogoldlabeling methodology using IgG and protein A-gold complex was implemented. T. spiralis larvae obtained from rat muscle were initially cultured in medium, and secreted excretory antigen was collected for 1 or 3 days. Purified antigenic protein was obtained from homogenized T. spiralis larvae. Rabbits were then immunized with 1 or 3 days secreted excretory protein and purified 45 kDa protein, and IgG was purified from collected serum. Serum, against infected antigen, collected from rat on 1 and 4 weeks after infection with T. spiralis larvae, and IgG was purified from collected serum. T. spiralis larvae were embedded in Lowicryl HM20 medium. Then they were finally treated with immunized IgG and protein A-gold complex (particle size; 15 nm) and observed under electron microscope. In T. spiralis larvae tissue, the tissue antigen reacted with rabbit IgC antigen Day 1 secreted excretory protein, infected antigenic protein and purified 45 kDa protein. But different distribution pattern of labeled gold particles were observed. When Day 1 secreted excretoy protein was used, gold particle labeling was observed specifically on the cuticle, basal layer, esophagus interstitial matrix (EIM) and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. In a separate group of tissue, the antigen reacted with rabbit IgG against Day 3 secreted excretory protein. Labeled gold particles were specifically distributed on the surface layer of cuticle, EIM and ${\alpha}_0$ granules of stichocyte of the worm. In case of using infected antigenic protein, gold particle labeling was specifically distributed on the cuticle and EIM of the worm. When purifed 45 kDa protein was used gold particle labeling was specifically distributed on the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. Therefore, excretory antigens appeared to originate from the cuticle and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte for the first day but the cuticle layer associated with globular proteins and ${\alpha}_0$ granules of stichocyte after 3 days and infected antigens appeared to originate from the cuticle for 1 and 4 weeks after infection. These results suggest that excretory and infection specific antigens are secreted into the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte and 45 kDa protein may be contained these specific antigens.

• Applied Microscopy
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• v.37 no.2
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• pp.93-102
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• 2007

Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.