• 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.

• Journal of fish pathology
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• v.10 no.2
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• pp.97-111
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• 1997

In the winter of 1995, mass mortality occurred in cultured flounder, Paralichthys olivaceus in Gurongpo, Kyoungbuk, Korea. From the observations of moribund and dead fish, parasitic ciliates, which were shown as white spots to the naked eye, were considered to be involved in the mass mortality. From heavily infected flounders, histopathological, morphological and biological characterization of these ciliates were carried out. In the histological observation, many ciliates were found under the epithelia of gill filaments and skin, and caused hyperplasia of epithelial and mucus cells at the infected areas. The ciliates found on the body surface, fins and gills were very similar to Cryptocaryon irritans. However the ciliates showed two different patterns of reproductian, i.e., typical form(palintomy)and atypical form(budding plus multiple fission) at $16^{\circ}C$ of water temperature. The occurrence ratio between typical and atypical form was about 3:2. Tomitogenesis takes 8-14 days in the typical and 13-15 days in the atypical form. In the viability test at different temperatures and salinities, the typical form died below 30‰ at $12^{\circ}C$, below 20‰ at $16^{\circ}C$, below 15‰ at $20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. On the other hand, the atypical form died below 20‰ at $12^{\circ}C$, below 15‰ at 16-$20^{\circ}C$, and below 25‰ at $24^{\circ}C$, respectively. The results suggested that the atypical has better viability at low salinity than that of the typical at low temperatures. In the excystment time and success rates of excystment according to temperatures, the typical form showed 8 days, 30% at $12^{\circ}C$ : 6.5 days, 50%, at $16^{\circ}C$ : 5.5 days, 75% at $20^{\circ}C$ : and 7 days, 10% at $24^{\circ}C$, respectively. On the other hand, the atypical form showed 15.5 days at $12^{\circ}C$ : 14 days, 76.6% at $16^{\circ}C$ : 12 days, 72.2% at $20^{\circ}C$ : 10 days 31.6% at $24^{\circ}C$, respectively. The results suggested that the atypical form had longer excystment time than that of the typical form at any temperature and showed better stability at low temperatures.

• Korean journal of food and cookery science
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• v.13 no.4
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• pp.440-447
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• 1997

The antibacterial effect of low concentrations of oregano (Origanum vulgare L.) in culture broth against Escherichia coli O157:H7 and Staphylococcus aureus 196E was tested at 35,5 and -20$^{\circ}C$. Tryptic soy broth (TSB) containing 0∼2% (w/v) of oregano was inoculated with 10$\^$6/∼10$\^$7/ CFU/$m\ell$ of E. coli or S. aureus and incubated at each temperature. The growth of E. coli was not inhibited at 0.1∼1.0% oregano and the growth occured at 2% oregano but only after a prolonged lag period. The death rate of E. coli after stationary phase was increased with increasing concentration of oregano in culture broth. The growth of S. aureus was inhibited with increasing concentration of oregano at 35$^{\circ}C$. Growth of S. aureus occured at the presence of 0.3∼0.5% oregano after a long lag period while the viability at 1.0∼2.0% was decreased during storage at 35$^{\circ}C$. During refrigerated storage at 5$^{\circ}C$, inhibition of E. coli or S. aureus was increased with the progress of time and increasing spice concentration. At the presence of 0.5∼2.0% oregano, E. coli and S. aureus were dead after 20 and 16 days of refrigerated storage, respectively. During frozen storage at -20$^{\circ}C$, the antibacterial activity of oregano against E. coli was increased with increasing storage time and spice concentration while the antibacterial activity against S. aureus was effective during the early period of storage, and no changes in the population of S. aureus occurred at different concentrations of oregano during frozen storage. Viable counts of E. coli were 1/3∼l/7 and S. aureus were 1/18∼l/46 of the control at 0.1% oregano in culture broth during frozen storage.

• The Korean Journal of Mycology
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• v.30 no.1
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• pp.50-55
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• 2002

Biological control against mushroom fly, Lycoriella mali, was performed by using Bacillus thuringiensis subsp. israelensis Bti-D and Bti-U, isolated from dead mushroom fly in oyster mushroom houses. Control values of the bacterial strains Bti-D and Bti-U against L. mali in bottle culture of oyster mushroom were 74.4% and 64.2%, respectively, and the value in small tray culture were 75.8% and 56.8%, respectively. In the experiment to develop the mass, cheap media for Bti-D and Bti-U isolates, the Biji broth (bean curd residue, called Biji in Korean language) was selected as a culture medium for an inexpensive and mass cultivation by the measurement of optical density of the two bacteria grown in the different media tested. Insecticidal effect of the formulation contained different ingredients that were prepared by using the Bti-D strain cultured in the Biji broth was tested in tray and bottle culture of oyster mushroom. The WCS formulation that contained corn starch as bio-gel (86.4%) was more effective to control the mushroom fly than living cells (69.1%) in bottle culture of oyster mushroom. Moreover, insecticidal effect of the WCS formulation was improved when water of pH 8 was used for dilution of the formulation. Effect of the WCS formulation using water of pH 8 and chemicals, Zuron (dimillin) W.P. on the control of mushroom fly and the productivity of oyster mushroom was investigated in tray culture of oyster mushroom. The Zuron W.P. was more effective to control the mushroom fly than the WCS formulation. However, compared with no treatment, the productivity of the mushroom treated with the WCS formulation was improved than that of the mushroom with Zuron W.P.

• Reproductive and Developmental Biology
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• v.28 no.3
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• pp.167-172
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• 2004

This study was performed to examine the correlations among dog sperm viabilities evaluated by flow cytometry, by microscopic evaluation (ME), by carbo-xifluorescein diacetate and propidium iodide (CFDA/PI) and by hypoosmotic swelling (HOS) test. Semen were collected from 5 dogs ranging in age from 2 to 4 years. Each ejaculate was divided into 3 aliquots and different proportions of freeze-killed cells were added to each aliquot (1:0, 1:1 and 1:3). In the other experiment, semen was extended with Sweden extender containing 5% glycerol and equex STM paste, and frozen using liquid nitrogen vapor. Fresh and frozen-thawed dog sperm viability were assessed by flow cytometry using PI staining method. The accuracy of flow cytometry was evaluated by comparing with other classic assessments, microscopic evaluation, epifluorescence microscopic analysis using CFDA/PI, and HOS test. High correlations of sperm viabilities were found among flow cytometry, epifluorescence evaluation, HOS test (p<0.01) in fresh semen. Especially, sperm viability assessed by HOS test was highly correlated with viability by flow cytometry in all the ratios of live and dead spermatozoa, 1:0, 1:1 and 1:3 (p<0.01). The viability evaluated by ME were significantly correlated with that by flow cytometry in ratios of 1:0 and 1:3 (p<0.05) however, there was no significance in ratio of 1:1. The viability evaluated by C/p were highly correlated with that by flow cytometry in ratio of 1:0 and 1:1 (p<0.01) and significantly correlated in ratio of 1:3 (p<0.05). In frozen-thawed spermatozoa, the viability determined by HOS test was considerably correlated with that by flow cytometry (p<0.01). There was significant correlation between the viabilities by ME and by flow cytometry (p<0.05). But the viability evaluated by CFDA/PI was not correlated with viability by flow cytometry. The result from this study validate the use of flow cytometry as a precise method for assessing the viability of fresh and frozen-thawed dog spermatozoa.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.19 no.4
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• pp.492-504
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• 2018

This study was conducted to investigate the effects of seaweed peeling (SP), glycolic acid peeling (GP) and general scrub (GS), which are widely known as cures for acne in both medicine and esthetics on the keratosis pilaris skin and provide basic data for a keratosis pilaris improvement program. For the experiment, subjects were categorized into control (GS) and experimental (GP and SP) groups, and tests were performed on arms and legs with relatively high keratosis pilaris symptoms (5 parts for each group) for 6 weeks. The keratin quantity, sebum content, moisture level and pigmentation were measured before and after (2, 4 and 6 weeks) the experiment and comparatively analyzed. The GP group showed an increase in moisture level (t=-4.064, p<0.01) but a decrease in pigmentation (t=3.536, p<0.01), while a decrease in keratin quantity (t=2.370, p<0.05) and pigmentation (t=4.017, p<0.01) was observed in the SP group and a decrease in keratin quantity (t=2.834, p<0.05) and an increase in moisture level (t=-7.589, p<0.001) was observed in the control group (GS). Additionally, the skin irritation reaction was lowest in the GS group. The SP group had the highest satisfaction with the improvement in response to keratosis pilaris care. When asked if they were willing to get the treatment with the same product, both SP and GP groups were high. In other words, keratosis pilaris care was needed in both experimental and control groups. Overall, the results of this study indicate that SP, GP and GS, which are commonly used in remedying acne, normalize turnover cycle by removing the dead cells from around the pores and improve keratosis pilaris symptoms by increasing moisture in the skin. Therefore, to improve keratosis pilaris skin, it is important to keep removing keratin and using a moisturizer that provides a skin barrier on a regular basis. The results presented herein will be useful as basic data for a keratosis pilaris improvement program.

• Parasites, Hosts and Diseases
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• v.27 no.3
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• pp.177-186
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• 1989

Observations were made on the differences in cell-mediated immune responses in the mice infected with strongly pathogenic Naegleria fewleyi ITMAP 359, weakly pathogenic Naegzeria jadini 0400, or non.pathogenic Naegleria gruberi EGB, respectively. Variations in cell-mediated responses and changes in antibody titers according to the duration after infection wore noted. Infections were done by dropping $5{\;}{\mu}l$ saline suspension containing $10{\times}10^4$ trophozoites cultured Bxenically in the CGVS medium into the right nasal cavity of ICR mice aging about 6~7 weeks, under the anesthesia by intraperitoneal injection of'secobarbital. Following infection, delayed type hypersensitivity(DTH) iesponses in the footpad and blastogenic responses of the mouse spleen cells using [$^3H$]-thymidine were observed on the day 1, 4, 7, 10 and 14 after infection. For the preparation of amoeba Iysates, each of cultured trophosoites were homogenized with an ultrasonicator, and centrifugated at 20,000 g. The supernatants of amoeba Iysates were used as the mitogen'and antigen for ELISA. Confanavalin A(Con. A) and lipopolysaccharide(LPS) were also used as mitogens in the blastogenic response. 1. The mice infected with N, fowleri showed the mortality rate of 75.7%. The rate was 6.2% for the N. jadini infected group, while no dead mouse was observed for N. gruberi infections. 2. In regard to DTH responses in the H. fewleri infected mice, the level increased in com- parison to the control group but declined after 7 days. An increase was also noted for the JV. jadini group after 1 day, but gradual decreases were observed through the infection period. In addition, no difference was noted between the N. gruberi infected and control groups. 3. Concerning the blastogenic response of the splenocytes, it increased after 10 days in the experimental group of N, fcwleri infection, but the differences ware not statistically significant compared with control group. It was evident that N. jadini group was not different from control group either, while there was a tendency of decrease in SV. gruberi infected group. In regard to the blastogenic response of the splenocytes by LPS, it was found that the N. fowlgri, N. jadini and N. gruberi infected groups had no differences from the control group. 4. The serum antibody titer of N. fcwleri and N. jadini infected mice increased from the day 7 and 14 after infection respectively, while the N. gruberi infected mice showed no increase. In summary of the results, it was observed that there were differences in the cell-mediated immune responses and serum antibody titers in the mice infected with strongly pathogenic JV. fowleri, weakly pathogenic N. jadini, or non.pathogenic N. gruberi, respectively.

• Journal of Life Science
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• v.26 no.1
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• pp.109-116
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• 2016

Ulmi cortex is the elm bark or root bark of Ulmus macrocarpa Hance and has been used as an ingredient of traditional medicine for anti-inflammatory, analgesic, anti-cancer and wound healing on both the East and the West. This study investigated whether the Ulmus macrocarpa Hance Water extract (UMWE) has the in vivo and in vitro immune activating effect. Animals were orally administrated for 14 days as follows: no treat group with distilled water, cyclophosphamide (CY) group with 120 mg/kg of CY, UMWE 100+CY group with 100 mg/kg of UMWE and 120 mg/kg of CY, UMWE 200+CY group with 200 mg/kg of UMWE and 120 mg/kg of CY, UMWE 100 group with 100 mg/kg of UMWE and UMWE 200 group with 200 mg/kg of UMWE. The immunosuppressive drug CY was intraperitoneally injected to induce immune suppression. Spleen indices showed small changes in CY injected groups but splenocyte indices showed greater decrease in the same groups. However, UMWE appeared to relieve CY’s immunosuppression. UMWE also delayed in vitro splenocyte death increasing its longevity. These data obtained by MTT assay and 7-amino-actinomycin D which stains preferentially dead than live cells. UMWE alone did not show cytotoxicity based on its apoptototic effect on splenocytes in vitro and in vivo. Splenic NK cell activity was maintained by UMWE under the presence of CY in vitro. The data indicated that UMWE protects splenocytes from the immunosuppressive drug CY under in vitro and in vivo conditions.

• Parasites, Hosts and Diseases
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• v.25 no.2
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• pp.168-180
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• 1987

This study was performed to observe histopathological changes and serological reactions in chronic anisakiasis of rabbits. Each rabbit was infected per os with 30 larvae of Anisakis type I. Their sera were collected chronologically and the rabbits were killed for histopathological examination, 3, 13, 20, 30, 60, 90 and 150 days after the infection. The results were summarized as below. 1. Most of the larvae were recovered from the stomach, but a few from the omentum, intestine, mesentery and abdominal wall. The recovery rates and distribution of worms by organ were not differed by duration of infection. 2. Histologically the lesion was abscess type on 13 days, i.e., the dead worms were surrounded by fibrinous exudate, histiocytes and thick zone of numerous inflammatory cells. After 30 days, histiocytes were found to invade the worms and the lesion was changing into abscessgranulomatous type. Also a calcified worm was found on the 30th day. After then the worms were observed to be dissolved slowly until 90 days. On 150 day, only one calcified worm was observed. 3. The levels of serum IgG antibody by ELISA reached their maximum 30 days after the infection. After then, it decreased slowly until 150 days after the infection. Above serological and histopathological findings indicated that antigenic stimulation from degenerating Anisakis larvae was the greatest during the first 30 days after infection. This period was corresponding with the beginning of worm resolution or calcification. Serologic test by ELISA would be a valuable tool for confirming chronic anisakiasis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.41 no.4
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• pp.341-350
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• 2015

Skin carrys out protective role against harmful outer environment assaults including ultraviolet radiation, heavy metals and oxides. Especially, ultraviolet-B (UVB) light causes inflammatory reactions in skin such as sun burn and erythma and stimulates melanin pigmentation. Furthermore, the influx of UVB into skin cells causes DNA damage in keratinocytes and dermal fibroblasts, inhibition of extracellular matrix (ECM) synthesis which leads to a decrease in elasticity of skin and wrinkle formation. It also damages dermal connective tissue and disrupts the skin barrier function. Prolonged exposure of human skin to UVB light is well known to trigger severe skin lesions such as cell death and carcinogenesis. Haloarcula vallismortis is a halophilic microorganism isolated from the Dead Sea, Its growth characteristics have not been studied in detail yet. It generally grows at salinity more than 10%, but the actual growth salinity usually ranges between 20 to 25%. Because H. vallismortis is found mainly in saltern or salt lakes, there could exist defense mechanisms against strong sunlight. One of them is generation of additional ATP using halorhodopsin which absorbs photons and produces energy by potential difference formed by opening the chloride ion channel. It often shows a color of pink or red because of their high content of carotenoid pigments and it is considered to act as a defense mechanism against intense UV irradiation. In this study, the anti-inflammatory effect of the halophilic microorganism, H. vallismortis, extract was investigated. It was found that H. vallismortis extract had protective effect on DNA damage induced by UV irradiation. These results suggest that the extract of halophilic bacterium, H. vallismortis could be used as a bio-sunscreen or natural sunscreen which ameliorate the harmful effects of UV light with its anti-inflammatory and DNA protective properties.