• Title/Summary/Keyword: 생체 외 실험

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Localization of Dendroaspis natriuretic peptide (DNP) in the rat salivary glands (흰쥐 타액선내에서의 DENDROASPIS NATRIURETIC PEPTIDE(DNP)의 분포)

  • Kim, Jae-Gon;Lee, Young-Soo;Baik, Byeong-Ju;Park, Byung-Keon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.3
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    • pp.447-463
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    • 2001
  • Dendroaspis natriuretic peptide (DNP), a fourth member of the natriuretic peptide isolated from the venom of the Dendroaspis angusticeps snake, has been reported to be present in human plasma and atrial myocardium and caused vasorelaxation and diuresis in experimental animals. However, it is uncertain whether they are present in peripheral organs other than the heart and its further physiological roles also remains to be clarified. To assess the possible physiological role of DNP in the salivary glands, I investigated the localization of DNP peptide in the rat salivary glands by immunohistochemistry and the binding sites for radiolabelled DNP in the rat salivary glands and oral mucosa using in vitro autoradiography. DNP immunoreactivity was widely distributed in the submandibular, sublingual and parotid glands, particularly in the ducts such as the intercalated and striated ducts, where atrial natriuretic peptide (ANP) was colocalized in consecutive sections, but not in acini. High density $^{125}I-DNP$ binding sites were localized in the epithelia of the tongue and hard palate, while low density binding sites for $^{125}I-DNP$ were also distributed in the submandibular, sublingual, and parotid glands. In the hard palate and tongue, the precise location of this binding was revealed on the basal and parabasal cells of the epithelia by emulsion microautoradiography. These results suggest that DNP may not only have a role in the salivary glands but also play a role in the regulation of growth in the oral epithelium, particularly in the hard palate and tongue.

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Effect of Thyroid Hormone on the Ischemia-Reperfusion Injury in the Canine Lung (갑상선 호르몬이 잡견 폐장의 허혈-재관류 손상에 미치는 영향)

  • 김영태;성숙환
    • Journal of Chest Surgery
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    • v.32 no.7
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    • pp.637-647
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    • 1999
  • Background: Ischemia-reperfusion injury is one of the major contributing causes of early graft failure in lung transplantation. It has been suggested that triiodothyronine (T3) may ameliorate ischemia-reperfusion injury to various organs in vivo and in vitro. Predicting its beneficial effect for ischemic lung injury, we set out to demonstrate it by administering T3 into the in situ canine ischemia-reperfusion model. Material and Method: Sixteen adult mongrel dogs were randomly allocated into group A and B. T3 $(3.6\mug/kg)$ was administered before the initiation of single lung ischemia in group B, whereas the same amount of saline was administered in group A. Ischemia was induced in the left lung by clamping the left hilum for 100 minutes. After reperfusion, various hemodynamic parameters and blood gases were analyzed for 4 hours while intermittently clamping the right hilum in order to allow observation of the injured left lung function. Result: Arterial oxygen partial pressure $(PaO_2)$ decreased 30 minutes after reperfusion and recovered gradually thereafter in both groups. In group B the decrease of $PaO_2$ was less marked than in group A. The recovery of $PaO_2$ was faster in group B than in group A. The differences between the two groups were statistically significant from 30 minutes after reperfusion $(125\pm34$ mmHg and $252\pm44$ mmHg, p<0.05) until the end of the experiment $(178\pm42$mmHg and $330\pm37$ mmHg, p<0.05). The differences in the arterial carbon dioxide pressure, airway pressure and lung compliance showed no statistical significance. The malondialdehyde (MDA) level, measured from the tissue obtained 240 minutes after reperfusion, was lower in group B $(0.40\pm0.04\mu$M) than in group A $(0.53\pm0.05\mu$M, p<0.05). The ATP level of group B $(0.69\pm0.07\mu$M/g) was significantly higher than that of group A $(0.48\pm0.07\mu$M/g, p<0.05). The microscopic exami nation revealed varying degrees of injury such as perivascular neutrophil infiltration, capillary hemorrhage and interstitial congestion. There were no differences in the microscopic findings between the two groups. CONCLUSION T3 has beneficial effects on the ischemic canine lung injury including preservation of oxygenation capacity, less production of lipid peroxidation products and a higher level of tissue ATP. These results suggest that T3 is effective in pulmonary allograft preservation.

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Effect of Inflammatory Responses to PLGA Films Incorporated Hesperidin: In vitro and In vivo Results (PLGA/헤스페리딘 함량별 필름에서 염증 완화 효과: In vitro, In vivo 결과)

  • Song, Jeong Eun;Shim, Cho Rok;Lee, Yujung;Ko, Hyun Ah;Yoon, Hyeon;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.37 no.3
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    • pp.323-331
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    • 2013
  • Hesperidin (Hes) has known to having some functions like protection of blood circulatory system, anti-tumor effect, antioxidant effect and anti-inflammatory effect. The goal of this study is to demonstrate the relationship between Hes and inflammatory through in vitro and in vivo studies using poly(lactic-co-glycolic acid) (PLGA) film including Hes as a tissue engineered scaffold. To confirm the proliferation of cells on fabricated scaffold, cells (RAW 264.7 and NIH/3T3) were seeded on PLGA/Hes film then analyzed with MTT and SEM at 1 and 3 days after seeding. The results from ELISA, RT-PCR, and FACS for anti-oxident and anti-inflammatory effect showed that inflammatory response of PLGA/Hes film decreased more than that of PLGA film. Also, in vivo result confirmed that inflammatory response by implanted PLGA/Hes film decreased more comparing with PLGA film. This is because of anti-inflammatory effect of Hes reducing induced inflammatory cell and accumulation of fibrous capsule. The results showed that PLGA/Hes film's capacity on reducing inflammatory is better than PLGA film because of Hes.

Interaction between Invertebrate Grazers and Seaweeds in the East Coast of Korea (동해안 조식성 무척추동물과 해조류 간 상호작용)

  • Yoo, J.W.;Kim, H.J.;Lee, H.J.;Lee, C.G.;Kim, C.S.;Hong, J.S.;Hong, J.P.;Kim, D.S.
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.12 no.3
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    • pp.125-132
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    • 2007
  • We estimated the distribution of predator-prey interaction strengths for 12 species of herbivores (including amphipods, isopods, gastropods, and sea urchins) and made a regression model that may be applicable to other species. Laboratory experiments were used to determine per capita grazing rate (PCGR; g seaweeds/individual/day). Relationship between the biomass of individual grazers and fourth-root transformed PCGR was fitted to power curve ($y=0.2310x^{0.3290}$, r=0.8864). This finding supported that the grazing efficiency was not even as individual grazers increase in size (biomass). Therefore, the biomass-normalized PCGR was estimated and revealed that smaller size herbivores were more effective grazers. Grazing impact considering density of each taxon was calculated. The sea hare Aplysia kurodai had greatest grazing impact on the seaweed bed and the sea urchin Strongylocentrotus nudus and S. intermedius were ranked in descending order of the impact. The amount of seaweed grazed by the amphipod Elasmopus sp. (>4,000 $ind./m^2$) and Jassa falcata (>2,000 $ind./m^2$) were 3.435 and $1.697mg/m^2/day$ respectively. The combined grazing amount of herbivores was $5,045mg/m^2/day$ in the seaweed bed. Although sea hare and sea urchin had strong impacts on seaweeds, the effects of dense, smaller species could not be seen as negligible. Surprisingly, the calculated grazing potential of sea urchins with a mean density of 3 $ind./m^2$ exceeded the mean production of seaweed cultured in domestic coastal waters in Korea (ca., 5 ton/ha). Small crustaceans were also expected to consume up to 16% of the seaweed production if their densities were rising under weak predation conditions. Considering that the population density of herbivores are strongly controlled by fish, human interference like overfishing may have strong negative effects on persistence of seaweeds communities.

The Role of c-Jun N-terminal Kinase in the Radiation-Induced Lung Fibrosis (방사선에 의한 폐 섬유화증에서 c-Jun N-terminal Kinase(JNK)의 역할)

  • Uh, Soo-Taek;Hong, Ki-Young;Lee, Young-Mok;Kim, Ki-Up;Kim, Do-Jin;Moon, Seung-Hyuk;Kim, Yong-Hoon;Park, Choon-Sik;Yeom, Uk;Kim, Eun-Suk;Choi, Doo-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.4
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    • pp.450-461
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    • 2001
  • Background : The underlying pathogenesis of radiation-induced lung fibrosis (RTLF) has not been very well defined. However, the role of TGF-$\beta$ in the generation of RTLF has been a major focus because there is an increase in the expression of both the TGF-${\beta}m$-RNA and its protein preceding RTLF lesions. The down stream signal after a TGF-$\beta$ stimulated lung fibrosis includes the activation of many mediators such as Smad and c-Jun N-terminal kinase (JNK) through TAK1. It is we hypothesized that JNK activation may play a pivotal role in RTLF pathogenesis through increased transcription of the fibrogenic cytokines. The present study evaluates JNK activity in alveolar macrophages after irradiation and the relationship between JNK activity and the amount of collagen in the lung tissues. Methods : C57BL/6 mice(20-25 gr, males) received chlorotetracycline(2g/L) in their drinking water 1 week prior to irradiation and continuously there after. The mice were irradiated once with 1400 cGy of $60CO{\gamma}$-ray over the whole chest. The cellular composition of the whole lung bronchoalveoalr lavage fluids(BALF), elastin expression in the lung tissues, the level of hydroxyproline in lung tissues, and an in vitro JNK assay was measured before irradiation and one, four, and eight weeks after irradiation (RT). Results : The volumes of BALF retrieved from instilled 4 mL of saline with 2% heparin were 3.7-3.8 mL for each group. The cell numbers were similar before($4.1{\times}10^4{\pm}0.5{\times}10^4/mL$) and 1 week($3.1{\times}10^4{\pm}0.5{\times}10^4/mL$) after RT. At four and eight weeks after RT, the cell number reached to $14.0{\times}10^4{\pm}1.5{\times}10^4mL$ and $10.0{\times}10^4{\pm}1.3{\times}10^4/mL$, respectively. There we no changes in the lymphocytes and neutrophils population observed in the BALF after RT. The H-E stain of the lung tissues did not show any structural and fibrotic change in the lung tissues at 4 and 8 weeks after RT. In addition, the amount of elastin and collagen were not different on Verhoeff staining of the lung tissues before RT to eight weeks after RT. The hydroxyproine content was measured with the left lung dissected from the left main bronchus. The lung were homogenized and hydrolyzed with 6 N Hel for 12 hours at $110^{\circ}C$ then measured as previously described. The content of hydroxyproline, standardized with a lung protein concentration, reached a peak 4 weeks after RT, and thereafter showed a plateau. AnIn vitro JNK assay using c-$Jun_{1-79}$-GST sepharose beads were performed with the alveolar macrophages obtained from the BAL. JNK activity was not detected prior to RT, However, the JNK activity increased from one week after RT and reached a peak four weeks after RT. Conclusion : JNK may be involved in the pathogenesis because the JNK activity showed similar pattern observed with the hydroxyproine content. However, it is necessary to clarify that the JNK increases the transcription of fibrogenic cyiokines through the transcription factor.

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The effect of denture adhesives on the retention of complete denture (의치 접착제가 총의치의 유지력에 미치는 영향에 관한 연구)

  • Yoon, Yu-Mi;Choi, Yu-Sung;Lee, Joon-Seok
    • The Journal of Korean Academy of Prosthodontics
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    • v.48 no.2
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    • pp.87-93
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    • 2010
  • Purpose: The purpose of this study was to compare the retention of the two denture adhesives with a silicone edentulous model and a resin denture base in vitro study. Material and methods: The experimental groups were used two denture adhesives and classified into 5 subgroups each. Subgroups were divided by the number of times the saliva sprayed. The control group was used synthetic saliva only. Dislodging tensile strengths were applied to the resin denture base using Instron in 3 directions ; vertical, lateral, and anterior-posterior. Results: The retention of two denture adhesives was increased significantly than saliva alone (P <.05). In each denture adhesives, the retention of saliva sprayed first was decreased than denture adhesive alone, but it didn't have significant differences. Whenever saliva sprayed repeated, the retention was decreased significantly than saliva sprayed first (P <.05). In each denture adhesives, vertical retentive force was highest than lateral and anterior-posterior retentive forces, and anterior-posterior retentive force was higher than lateral retentive force. This results were significantly different (P < .05). Significant differences of the retentive ability among two denture adhesives were not observed. Conclusion: From the results, use of the denture adhesives resulted in improved retentive ability of denture. Especially retentive force was highest in vertical direction.

Preparation and Release Behavior of Ipriflavone-Loaded PLGA Microsphere for Tissue Engineered Bone (이프리플라본을 함유한 생분해성 PLGA 미립구의 제조 및 조직공학적 골재생을 위한 영향평가)

  • So, Jung-Won;Jang, Ji-Wook;Kim, Soon-Hee;Kim, Geun-Ah;Choi, Jin-Hee;Rhee, John-M.;Son, Young-Suk;Min, Byoung-Hyun;Khang, Gil-Son
    • Polymer(Korea)
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    • v.33 no.1
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    • pp.26-32
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    • 2009
  • The aim of this research was to prepare microparticulate systems based on poly (lactide-co-glycolide)(PLGA) for the local release of ipriflavone in order to reduce bone loss. We developed the IP loaded PLGA microspheres using relatively simple oil-in-water(O/W) solvent evaporation method. HPLC was used to perform the in vitro release test of IP and morphology of cell attached on the micro-spheres was investigated using SEM. Cytotoxicity was assayed by cell counting kit-8 (CCK-8) test. Osteogenic differential cells were analyzed by ALP activity. Through RT-PCR analysis, we observed osteocalcin, ALP, and Type I collagen mRNA expression. The release of IP in vitro was more prolonged over 42 days and IP/PLGA microspheres showed the improvement on the cell proliferation, ALP activity and RT-PCR comparing with control (only PLGA). This initial research will be used to direct future work involved in developing this composite injectable bone tissue engineering system.

INHIBITORY EFFECT OF Er:YAG LASER ON THE GROWTH OF STREPTOCOCCUS MUTANS (Er:YAG 레이저 조사가 Streptococcus mutans의 증식억제에 미치는 효과)

  • Song, Gwang-Chul;Lee, Chang-Seop;Lee, Sang-Ho;Lee, Nan-Young
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.1
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    • pp.15-24
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    • 2003
  • The purpose of this study is to investigate the sterilization effect of Er:YAG laser against the intraoral acid producing bacterium, S. mutans, by irradiating the culture solution containing S. mutans KCTC 3065 with Er:YAG laser having a $650{\mu}m$ diameter beam through the non-contact method. We obtained the following results after examining the temperature changes of the culture solution, numbers of bacterial colonies, and acid-producing ability and attaching ability on teeth by measuring the amount of extracellular polysaccharide produced by S. mutans. The number of bacterial colony was decreased in $10{\mu}l$ culture solution irradiated with laser in overall compared to the control solution. The number decreased as the irradiation intensity and pulse repetition rate were larger and as the exposure time was increased. However, it did not change significantly in $100{\mu}l$ culture solution compared to the control solution. Although the acid-producing ability of S. mutans was inhibited for a certain duration after laser irradiation in 10r1 bacterial culture solution, it did not change in $100{\mu}m$ solution compared with the control solution. The amount of extracellular polysaccharide synthesized by S. mutans was partially decreased through laser irradiation in $10{\mu}m$ culture solution but did not change in $100{\mu}m$ culture solution. Based on these findings, we concluded that Er:YAG laser has an sterilization effect on S. mutans in which we presume that the mechanism is through the heat effect rather than the mechanical effect from development of ultrasound.

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Tissue Engineered Cartilage Formation on Various PLGA Scaffolds (PLGA 종류와 담체의 형성 방법에 따른 인간의 조직공학적 연골형성)

  • 김유미;임종옥;정호윤;박태인;백운이
    • Journal of Biomedical Engineering Research
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    • v.23 no.2
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    • pp.147-153
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    • 2002
  • The purpose of this study was to evacuate the effect of different types of Poly(lactic-co-glycolic acid) (PLGA) scaffolds on the formation of human auricular and septal cartilages. All of the scaffolds were formed in a tubular shape for potential application for artificial trachea or esophagus with either 110,000 g/mol PLGA. 220,000 g/mol PLGA. or a combination of both. In order to maintain the tubular shape in vivo, two methods were used. One method was inserting polyethylene tube at the center of scaffolds made of 110,000 g/mol PLGA. The other method involved combination of the two different molecular weight PLGA's. The inner surface of tubular shaped scaffold made with 110,000 g/mol PLGA was coated with 220,000 9/mol PLGA to give more mechanical rigidity. Elastic cartilage was taken from the ear of a patient aged under 20 nears old and hyaline cartilage was taken from the nasal septum. The chondrocytes were then isolated. After second passage, the chondrocytes were seeded on the PLGA scaffolds followed by in vitro culture for one week. The cells-PLGA scaffold complex were implanted subcutaneously on the back of nude mice for 8 weeks. The tissue engineered cartilages were separated from nude mice and examined histologically after staining with the Hematoxylin Eosin. The morphology of the scaffolds were examined by scanning electron microscopy. The pores were well formed and uniformly distributed in the various PLGA scaffolds. After 8 weeks in vivo culture, cartilage was well formed with 110,000 g/mol PLGA. however lumen had collapsed. In contrast. a minimal amount of neocartilage was formed with 220,000 g/mol PLGA, while the architecture of scaffold and lumen were well preserved. Elastic cartilage formed more neocartilage than hyaline. Hyaline and elastic neocartilage were well formed on 110,000 g/mol PLGA with the polyethylene tube, exhibiting mature chondrocytes and preservation of the tubular shape. It was found that 110,000 g/mol PLGA was more appropriate for cartilage formation but higher molecular weight polymer was necessary to maintain the three dimensional shape of the scaffold.

Differentiation and Apoptosis of the Mammalian Embryo and Embryonic Stem Cells(ESC): I. Establishment of Mouse ESC and Induction of Differentiation by Reproductive Hormones (포유동물의 배아 및 기간세포의 분화와 세포사멸 기작: I. 생쥐 배아줄기세포의 확립과 분화유도에 미치는 생식호르몬의 영향)

  • 성지혜;윤현수;이종수;김철근;김문규;윤용달
    • Development and Reproduction
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    • v.6 no.1
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    • pp.55-66
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    • 2002
  • Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

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