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Effects of Electrical Stimulation on the Nucleolar Organizer Region in Keratinocyte of Wounded Skin in Rat (전기자극이 흰쥐 창상 표피세포의 핵소체형성부위에 미치는 영향)

  • Lee Jae-Hyoung;Jekal Seung-Joo;Park Seung-Teack
    • The Journal of Korean Physical Therapy
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    • v.12 no.1
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    • pp.1-7
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    • 2000
  • The purpose of this study was to determine the effect of microamperage electrical stimulation on the number of argyrophilic nucleolar organizer region (AgNOR) in rat skin. Twenty four male Sprague-Dawley rats were divided into electrical stimulation and control group. Bach animals hair on the back was removed. The electrical stimulation group received an positive rectangular positive electrical stimulation with $500{\mu}A$, while the control group was given the same treatment without electricity. The rats were sacrificed at 4 and 7 day of stimulation, respectively. The biopsy specimens were fixed in formalin, embedded in paraffin and stained with silver nitrate. The AgNOR were counted using a light microscope and computerized image analysis system and calculated as the mean number of AgNOR per nucleus in the epidermal keratinocyte. In control skin, the mean AgNOR count of epidermal keratinocyte at 4 and 7 day were 1.67 and 1.72, whereas electrical stimulated rat had mean AgNOR counts of 2.0 and 2.14, respectively. A Student's t-test showed a significantly higher mean AgNOR number at 4 ana 7 day in the electrical stimulated rats than control rats (p<0.05). The microamperage electric current stimulation increased the epidermal AgMOR expression in incisional wound skin. These results suggest that the microamperage electrical stimulation may promote migration and proliferative activity of epidermal keratinocyte in surgical wound.

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Numerical and morphologic changes of ovarian follicles in each stage of estrus cycle in rats (Rat의 성주기에 따른 난포의 수와 형태변화)

  • Lee, Yoi-joo;Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.455-462
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    • 1999
  • This study was designed to investigate the number of the growing and mature follicles in each stage of estrus cycle in mature rats. Eighteen mature rats(Sprague-Dawley, initially 190~230gm) were randomly alloted into 4 groups(proestrus, estrus, metestrus, and diestrus) according to estrus cycles. The uteri and ovaries of rats were collected and then alternative sections of paraffin embedding ovaries were stained with H-E. Numbers of large, middle and small follicles or only large and middle follicles from secondary and tertiary follicles were investigated by LM photography of preparations. Small follicles were defined as secondary follicles with 2~5 cell layers of granulosa cells surrounding the oocyte, and middle follicles were defined as secondary follicles with more than 5 cell layers or with early signs of antral cavity or with more than one small cleft on either side of the oocytes and large follicles were defined as tertiary follicles with a single medium or large antral cavity. The number of follicles in a pair ovary per rat was appeared to be ranged from 207 to 370 and the mean number of these follicles was $270.4{\pm}52.6$ and the mean number of follicles per ovary was $134.9{\pm}32.0$. The mean number of large, middle and small follicles per ovary was appeared to be $16.4{\pm}4.4$($12.2{\pm}3.3%$), $36.2{\pm}8.6$($26.8{\pm}6.4%$), and $82.7{\pm}24.0$($61.3{\pm}17.8%$), respectively. The mean number of large and middle follicles in each stage group of estrus cycle was appeared to be $17.8{\pm}2.1$ and $38.3{\pm}7.4$ at proestrus stage group, $15.7{\pm}5.2$ and $38.0{\pm}10.0$ at estrus stage group, $16.5{\pm}3.5$ and $33.8{\pm}7.0$ at metestrus stage group, $16.7{\pm}5.8$ and $29.7{\pm}5.5$ at diestrus group, respectively. In histological findings of large follicles during each estrus cycle, the large follicles in proestrus group contain single small antrum, thick granulosa cell layers, and were $300{\sim}500{\mu}m$ in diameter and were growing follicles with PCNA-positive cells in the granulosa cell layers, and other luteinizing follicles of proestrus cycle stage were decreased in size and were thicker in wall thickness and more luteinized than those in metestrus and diestrus stage groups. The large follicles in estrus stage group contain thick granulosa cell layers and nonprominent cumulus-oocyte complexes in antrum, and were $400{\sim}700{\mu}m$ in diameter and were growing follicles with PCNA-positive cells in the granulosa cell layers. The large follicles in metestrus and diestrus stage groups contain enlarged antrums, thinner layers of walls and prominent cumulus-oocyte complexes, and were $700-950{\mu}m$ in diameter, and were nongrowing follicles without PCNA-positive cells or another large follicles contain cells with dark stainability and distinct boundary.

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Immunohistochemical study on the atretic and the growing follicles after experimental superovulation in rats 2. Atresia and growing of follicles (과배란 유기된 rat 난소에 퇴축난포와 성장난포에 대한 면역조직화학적 연구 2. 동원된 난포의 퇴축과 성장에 대하여)

  • Kwak, Soo-dong;Koh, Phil-ok;Kim, Chong-sup
    • Korean Journal of Veterinary Research
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    • v.37 no.1
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    • pp.79-86
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    • 1997
  • This study was designed to investigate the effects of superovulation on the growing and mature follicles following gonadotrophin treatments in mature rat by immunohistochemical methods. Eighteen mature rats (Sprague-Duwely, 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone(FSH) / rat, and third PMS and HCG-treated group was injected intramuscularly with 20~25IU of pregnant mare serum(PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotropin(HCG) / rat. Half the number of rats were administrated intraperitoneally with bromodeoxyuridine(Brdur, 0.2mg/gm BW once) at 2 hours before exanguination and the remainder of rats were sacrified without Brdur administration. The investigation by immunohistochemical methods using paraffin sections of ovaries was performed by using anti-Brdur antibody and PCNA(proliferating cell nuclear antigen) antibody for labeling proliferating cells in follicles. In immunohistochemical findings, follicles squeezed by peripheral corpus luteum or follicles large follicles with loosly and irregularly distributed granulosa cells and although with compacted granulosa cells, middle follicles with dilated round or oval follicular antrum were confirmed as atretic follicles. The proportions of atretic follicles in control group were 29.8%, 21.7% and 14.2% respectivley at large, middle and small follicles and mean proportions of these all 3 grade follicles were 26.7%. The proportions of atretic follicles in FSH-treated group were 35.4%, 24.9% and 10.4% respectively at large, middle and small follicles and mean proportions of these all 3 grade follicles were 28.1%. The proportions of atretic follicles in PMS and HCG-treated group were 44.7%, 24.0% and 12.7% respectively at large, middle and small follicles, and mean proportions of these all 3 grade follicles were 29.7%. The above findings reveal that the group with higher proportion of atretic follicles were ordered as large, middle and small follicles in size, and these proportions were increased in hormone treated two groups with more number of more growing and mature follicles when compared with control group.

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Influence of Coating Materials and Emulsifiers on Nanoparticles in Manufacturing Process (코팅물질과 유화제가 나노입자 제조 및 안정성에 미치는 영향)

  • Kim, Byeong-Cheol;Chun, Ji-Yeon;Park, Young-Mi;Hong, Geun-Pyo;Lee, Si-Kyong;Choi, Mi-Jung
    • Food Science of Animal Resources
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    • v.32 no.2
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    • pp.220-227
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    • 2012
  • The objective of this study was to investigate the influence of emulsion processing with various homogenization treatments on the physical properties of nanoparticles. For the manufacturing of nanoparticles, by taking the emulsion-diffusion method, various coating materials, such as gum arabic, hydroxyethyl starch, polycarprolactone, paraffin wax, ${\kappa}$-carrageenan and emulsifiers like Tween$^{(R)}$60, Tween$^{(R)}$80, monoglyceride and Pluronic$^{(R)}$F68, were added into the emulsion system. Furthermore, the various speeds (7,000 rpm to 10,000 rpm), and times (15 s to 60 s) of homogenization were treated during the emulsion- diffusion process. NEO II homomixer was the most effective homogenizer for making nanoparticles as 51 nm ($D_{10}$) and 26 nm ($D_{50}$). To manufacture smaller nanoparticles, by using NEO II homomixer, 10,000 rpm of agitation speed, polycaprolactone as coating material, and Pluronic$^{(R)}$F68 as an emulsifier were the optimum operating conditions and components. For the stability of nanoparticles for 7 days, $20^{\circ}C$ of storage temperature was appropriate to maintain the particle size. From these results, the type of homogenizer, homogenization speed, homogenization time and storage temperature could affect the particle size. Moreover, type of coating materials and emulsifier also influenced the size and stability of the nanoparticles.

Environmental-Friendly Control of the Ginseng Stem Fungus Gnat (Phytosciara procera, Diptera) in the Ginseng Field (인삼재배지에서 인삼줄기버섯파리(Phytosciara procera)의 친환경 방제)

  • Seo, Mi-Ja;Shin, Hyo-Seob;Gawk, Chang-Soon;Jo, Shin-Hyuk;Cho, Hye-Sun;Shin, Jung-Sup;Cho, Dae-Hui;Youn, Young-Nam;Yu, Yong-Man
    • The Korean Journal of Pesticide Science
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    • v.15 no.4
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    • pp.471-478
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    • 2011
  • The ginseng stem fungus gnat, Phytosciara procera, overwintered in old stems and roots of ginseng as a larvae from late September to early May in next year. In the late Spring they become pupa then emerged. Therefore, the first control period has to be set between late May and early June. It is a same period of elimination of ginseng flower stalks. Two or three times pest control might be needed per month for controlling adult of P. procera. Otherwise, the overwintered form as larva in old stem by removal of old stems of ginseng, the population of first generation of P. procera adults could be reduced. However, it seemed that the removal of old stems was not necessarily correlated with the population of P. procera adults in the next generation. Three kinds of environmental-friendly control agents that were mainly contained sophora extract, showed the highly insecticidal activities against larva of P. procera. Among them, one agent containing sophora extract and paraffin oil showed an ovicidal action as well as oviposition repellency.

Preparation and Effect of Eudragit E100 Microcapsules Containing Grapefruit Seed Extract on Kimchi (자몽씨 추출물을 함유한 Eudragit E100 미세캡슐의 제조 및 김치에 대한 영향)

  • 김한수;정성기;조성환;구재관;이승철
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.8
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    • pp.1239-1244
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    • 2003
  • Microcapsules were prepared by coacervation method using acetone/liquid paraffin system to control the ripening of kimchi. Eudragit E100, which was soluble at below pH 5.0 in aqueous solution, was used to make microcapsules to be sensitive to acidity of kimchi. The microcapsules with Eudragit E100 containing grapefruit seed extract (GFSE) showed the highest yield of 92.13%, the size of microcapsules was decreasing as increasing the amount of aluminium stearate, a dispersing agent. Morphology of the microcapsules determined by scanning electron microscopy showed spherical forms. GFSE, encapsulated antimicrobial agents, was quickly released at acidic buffer (pH 4,5,6) within 1 storage day. However, 70% of encapsulated GFSE in Eudragit E100 microcapsules was continuously released at pH 7 till 3 days, and it was sustained till 9 days. Characteristics of kimchi containing microcapsules of GFSE were analysed with ripening period. Decease of pH in kimchi was retarded with the added GFSE microcapsules till 2 days of fermentation, but GFSE did not affect pH in kimchi after 3 days. Total numbers of microorganisms and lactic acid microorganisms in kimchi were decreased with increasing the amount of the added GFSE microcapsules, however, the effect of controlled released GFSE from pH sensitive Eudragit E100 microcapsules was hard to detect. These results suggest the possibility of pH sensitive microcapsules for high qualify of kimchi.

Characterization of Monoclonal Antibodies against Human Leukocyte Common Antigen (CD45)

  • Shin, Hyang-Mi;Cho, Woon-Dong;Lee, Geon-Kook;Lee, Seon-Hwa;Lee, Kyung-Mee;Ji, Gil-Yong;Yoon, Sang-Soon;Koo, Ji-Hae;Lee, Ho-Chang;Lee, Ki-Hyeong;Song, Hyung-Geun
    • IMMUNE NETWORK
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    • v.11 no.2
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    • pp.114-122
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    • 2011
  • Background: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. Methods: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. Results: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). Conclusion: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen.

THE EFFECTS OF TGF-$\beta$1 ON THE BONE REGENERATION IN GUINEA PIG MANDIBLE (형질변형성장인자가 손상골 재생능에 미치는 영향에 관한 실험적 연구)

  • Lee Byung-Do;Hwang Eui-Hwan;Lee Sang-Rae;Hong Jung-Pyo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.25 no.2
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    • pp.423-435
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    • 1995
  • The purpose of this study was to observe the effect of TGF-βl, which promotes differentiation and proliferation of osteoblasts, on bone regeneration. Experimental bone defects that measured 3 mm in diameter were created on the mandibles of guinea pig by removal of bone with the use of trephine burs. In one side of mandibular body, the experimental groups, bone defects were grafted with Biogran(Orthovita Co., U.S.A) and TGF-β1(R&D System Co., U.SA). In the remaining side of the mandiblar body, the control groups, bone defects were grafted with only Biogran. Guinea pigs in the control and experimental groups were serially terminated by fours on the 3 days, the 1 week, the 2 weeks, the 3 weeks, and the 4 weeks after experiment, and both sides of the mandibular bodies were removed and fixed with 10% neutral formalin. They were decalcified and embedded in paraffin as using the usual method. The specimen sectioned and stained with hematoxylin and eosin. Also, they were radiographed with a soft X -ray apparatus. The obtained results were as follows; 1. Hemorrhagic condition, observed in the granulation tissues, disappeared on the 1 week after experiment in both groups, and more prominent in the experimental group. The granulation tissues of the experimental group had larger number of cells than those of the control group. 2. Osteoblastic differentiation in the margin of grafted material and adjacent bone was observed on the 1 week after experiment in both groups. Also, bone formation was observed in immature form on the 1 week after experiment. and more prominent in the experimental group. 3. In the polarizing microscopic examination, bone matrix was very loose on the 1 week after experiment, but increase in density with time, and more prominent in the experimental group. 4. In the microradiographic examination, newly formed bone was observed in the experimental group on the 2 weeks after experiment, and this was observed earlier than in the control group. Newly formed bone was increased with time and defected area was markedly decreased on the 4 weeks after experiment.

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Expression of vascular endothelial growth factor receptors in tumor and stromal cells of tongue squamous cell carcinoma

  • Park, Bong-Wook;Byun, June-Ho;Hah, Young-Sool;Kim, Deok-Ryong;Chung, In-Kyo;Kim, Jong-Ryoul;Kim, Uk-Kyu;Park, Bong-Soo;Kim, Gyoo-Cheon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.1
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    • pp.11-19
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    • 2007
  • This study was to evaluate the expression of vascular endothelial growth factor receptors (VEGFRs) in tumor and stromal cells of tougue squamous cell carcinoma (SCC). We also wanted to characterize the differences, from the angiogenic aspect, between cancer-associated stromal cells and non-malignant stromal cells. Paraffin-embedded tumor specimens from eleven patients with tongue SCCs were studied. Immunohistochemical staining for VEGFR-1,-2, and -3 was performed on the tumor cells, stromal fibroblasts and tumor-associated macrophages of the specimens. The expression of all 3 receptors was detected in the tumor cells themselves of the biopsy specimens. All 3 receptors were also expressed on stromal cells, except that VEGFR-2 was not expressed in stromal fibroblasts. In radical excision specimens, the staining intensity for VEGFR-1, -2 in the tumor cells and VEGFR-1,-3 in the tumor-associated macrophages was significantly lower than that in the biopsy specimens (P < 0.05). By using the general marker of fibroblast and macrophage, 5B5 and CD68, respectively, we performed double immunofluorescence staining for 5B5 and each VEGFR in the stromal fibroblasts and for CD68 and each VEGFR in the tumor-associated macrophages of the radical excision specimens. We used 4 cases of fibroma and 4 cases of chronic inflammation tissue as the controls. It was found that only each marker was expressed in the control group, however, 5B5/VEGFR-1 and 5B5/VEGFR-3 in the stromal fibroblasts, and CD68/VEGFR-1 and CD68/VEGFR-3 in the tumor-associated macrophages were double stained in the radical excision specimens. Although our study used small number of specimens, the results of our study showed that in tongue SCC, in association with the angiogenesis, the stromal cells showed the activated phenotype and this was different from the nonmalignant stromal cells.

EFFECTS OF ELECTRICAL STIMULATION ON THE NORMAL PERIODONTIUM (전기자극이 정상 치주조직에 미치는 영향)

  • Lim, Kyung-Seok;Kwon, Young-Hyuk;Lee, Man-sup;Park, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.32 no.1
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    • pp.89-112
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    • 2002
  • The earliest reports of the use of electrical energy to directly stimulate bone healing seem to be in 1853 from England, the techniques involved the introduction of direct current into the non-united fracture site percutaneously via metallic needles, with subsequent healing of the defect. One endpoint of the periodontal therapy is to generate structure lost by periodontal diseases. Several procedural advances may support regeneration of attachment, however, regeneration of alveolar bone does not occur consistently. Therefore, factors which stimulate bone repair are areas for research in periodontal reconstructive therapy. Effects of cytokines or growth factors on bone repair are examples of such areas. Another one is electrical current which occurs in bone naturally, so that such bone may be particularly susceptible to electrical therapy. The purposes of this study were to observe the effects of electrical stimulation on the normal periodontium, to determine whether the electricity is the useful means for periodontal regeneration or not. Forty rats weighted about 100 gram were used and divided into 4 groups, the first group, there was no electrical stimulation with the connection of electrodes only. In the second group, there was stimulated by the 10 mA during 10 minutes per a day, in the third group was stimulated by the 25 mA , and the fourth by the 50 mA. At 3, 5, 10 and 15 days post-appliance , two rats in each group were serially sacrificed. and the maxillae and the mandible processed to paraffin, and the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows : 1. There was the distinct reversal line on the lingual alveolar crest, whereas a little changes in the labial alveolarcrest to the duration and amount of currents. 2. In 50 mA group, the cells were highly concentrated at the apex of anterior teeth, and was observed the necrotic tissue. In posterior root apex, the hypercementosis was appeared, and newly formed cementum layer has been increased continuously with the time. 3. The periodontal ligament fiber and Sharpey's fiber were arranged in order, and the bone trabeculae were increased as the experiment proceeded by, relatively the bone marrows were decreased. 4. In the pulp tissue, the blood vessels were increased with blood congestion in the experimetal specimens remarkably, and the dentinal tubules were obstructed . 5. The osteoblasts in alveolar bone proper had been showed highly activity, and also observed the formation of bone trabeculea. In the conclusion, it was suggested that the electrical stimulation has influence on the periodontium and the pulp tissue. However, there might be the injurious effects.