• Title/Summary/Keyword: Extracellular

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The Action and Pacemaker Potential in the Frog Truncus Arteriosus (개구리 대동맥의 활동전압 및 Pacemaker 전압에 관한 연구)

  • Earm, Yung-E;Sung, Ho-Kyung
    • The Korean Journal of Physiology
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    • v.16 no.2
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    • pp.119-128
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    • 1982
  • The frog truncus arterious were studied with conventional glass microelectrode technique in order to elucidate the underlying mechanism of spontaneous pacemaker activity. The analyses were focussed on the ionic nature of pacemaker current by changing the concentrations of extracellular $K^+$ and, $Na^+$, or by using blockers of K- and Ca-current and chronotropic transmitters. 1) The action potential of the spontaneously active truncus arteriosus has some characteristic feature of maximal distolic potential ranged from -65 to -75 mV, resting potential from -45 to -50 mV and overshoot voltage about +30 mV, respectively. Duration of the action potential taken from rapid upstroke to maximal diastolic potential was about 600 msec. Usual discharge rate was $25{\sim}30/min$ at room temperature $(18{\sim}20^{\circ}C)$. 2) The sensitivity of the resting membrane potential to change extracellular potassium concentrations $(0{\sim}12\;mM)$ was relatively low. Transient hyperpolarization was appeared in the 12 mM K Ringer after 10 min exposure to 0 mM K and it could be related to Na-pump reactivation by high potassium. 3) Reduction of extracellular sodium concetrations diminished the amplitude and frequency of the action potential. In Ringer solution containing 30% Na (substituted by equimolar Tris), spontaneous activity stopped but reappeared as very slow and small action potential. There was no spotaneous activity in zero Na Ringer solution. 4) Caesium(10 mM), K-current blocker decreased the frequency of the action potential and also pacemaker depolarization. Manganese (2 mM) known to be Ca-current antagonist, blocked spontaneous activity completely. 5) Adrenaline and acetylcholine had no chronotropic effect. But adrenaline increased the duration of plateau phase and the magnitude of the action potential in the follower cell. It is concluded that K-, Na-and Ca-current components are involved in the genesis of spontaneous activity of the frog truncus arteriosus like cardiac pacemaker tissues. But the insensitivity of truncus arteriosus to adrenaline and acetylcholine indicates that there are some different control mechanisms of spontaneous rhythm in two tissues.

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pH-mediated Regulation of Pacemaker Activity in Cultured Interstitial Cells of Cajal

  • Kim, Byung-Joo;Lee, Jae-Hwa;So, In-Suk;Kim, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.10 no.1
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    • pp.7-11
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    • 2006
  • Interstitial cells of Cajal (ICCs) are pacemakers in gastrointestinal tracts, regulating rhythmicity by activating nonselective cation channels (NSCCs). In the present study, we investigated the general characteristics and pH-mediated regulation of pacemaker activity in cultured interstitial cells of Cajal. Under voltage clamp mode and at the holding potential of -60 mV, the I-V relationships and difference current showed that there was no reversal potential and voltage-independent inward current. Also, when the holding potentials were changed from +20 mV to -80 mV with intervals of 20 mV, there was little difference in inward current. In pacemaker activity, the resting membrane potential (RMP) was depolarized (In pH 5.5, $23{\pm}1.5$ mV depolarized) and the amplitude was decreased by a decrease of the extracellular pH. However, in case of increase of extracellular pH, the RMP was slightly hyperpolarized and the amplitude was decreased a little. The melastatin type transient receptor potential (TRPM) channel 7 has been suggested to be required for intestinal pacemaking activity. TRPM7 produced large outward currents and small inward currents by voltage ramps, ranging from +100 to -100 mV from a holding potential of -60 mV. The inward current of TRPM7 was dramatically increased by a decrease in the extracellular pH. At pH 4.0, the average inward current amplitude measured at -100 mV was increased by about 7 fold, compared with the current amplitude at pH 7.4. Changes in the outward current (measured at +100 mV) were much smaller than those of the inward current. These results indicate that the resting membrane potential of pacemaking activity might be depolarized by external acidic pH through TRPM7 that is required for intestinal pacemaking activity.

Comparative Analysis of the Extracellular Matrix Composition in Proliferating and Involuted Infantile Hemangiomas

  • Park, Hyochun;Park, Hannara;Chung, Ho Yun;Teresa, MO;Waner, Milton
    • Archives of Plastic Surgery
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    • v.42 no.5
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    • pp.544-551
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    • 2015
  • Background Changes in the composition of the extracellular matrix (ECM) occur between the proliferating and involuted phases of infantile hemangiomas (IH), and are associated with angiogenic growth. We examined the composition of the ECM in proliferating and involuted IHs and assessed correlations between the composition of the ECM and whether the IH was in the proliferating or the involuted phase. Methods We evaluated IH samples from a cohort of patients who had five proliferating IHs and five involuted IHs. The following ECM molecules were analyzed using enzyme-linked immunosorbent assays and immunohistochemistry: laminin, fibronectin, collagen type I, collagen type II, and collagen type III. Results The involuted IHs had higher levels of deposition of collagen type III than the proliferating IHs. The median values (interquartile ranges) were 1.135 (0.946-1.486) and 1.008 (0.780- 1.166) (P=0.019), respectively. The level of laminin was higher in involuted IHs than in proliferating IHs, with median values (interquartile ranges) of 3.191 (2.945-3.191) and 2.479 (1.699- 3.284) (P=0.047), respectively. Abundant collagen type III staining was found in involuted IHs. Laminin ${\alpha}4$ chain staining was clearly present within the basement membrane adjacent to the blood vessels, and was significantly more intense in involuted IHs than in proliferative IHs. Conclusions Involuted hemangiomas showed extensive deposition of collagen III and laminin, suggesting that differences in the composition of the ECM reflect stages of the development of IHs. This pattern may be due to the rapid senescence of IHs.

Electron Microscopic Study on the Replcation of Hantaan Virus in Vero-E6 Cells (Vero-E6 세포에서 한탄바이러스의 증식에 관한 전자현미경적 연구)

  • Park, Kyung-Hee;Seong, In-Wha
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.201-209
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    • 1999
  • Results of the studies on the morphologic and molecular biologic characteristics of Hantaan virus (HTNV), one of the etiologic agents of Hemorrhagic fever with renal syndrome (HFRS), revealed that HTNV was a member of Family Bunyaviridae and its RNA divided into three segments. And the nucleotide sequences of these segments also were known and the differences in nucleotide sequences of HTNV from other members of genus Hantavirus were clearly evaluated. But the morphorgenesis, pathogenesis of HFRS and the replication time had not been clearly determined. In this study, to estimate the replication time of HTNV in Vero E-6 cells, Vero cells were infected with HTNV 76/118 strain, and cells were harvested from two hours post-infection up to 24 hours at two hours-intervals. Harvested cells were treated with ordinary techniques for electron microscopy and immune-electron microscopy. And then thin sections were observed under transmission electron microscope. HTNV particles were not found in the cytoplasm and in the extracellular space between $2{\sim}8$ hours after inoculation of virus, but virus particles were observed in extracellular space near the cell membrane of Vero-E6 cells 10 hours after infection. In immune electron microscopy, mature HTNV particles in extracellular spaces and immature virus labelled with gold particles in the cytoplasm of Vero E-6 cell 10 hours after infection of HTNV could be seen. This results suggest that the replication time of HTNV might be about 10 hours.

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Stimulation of the Extracellular Matrix Production in Dermal Fibroblasts by Areca catechu Extract (진피섬유모세포에서 대복피추출물의 세포외기질 합성 촉진 효과)

  • Lee, Min-Ho;Kim, Hyung-Jin;Jung, Hyun-Ah;Lee, Young-Keun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.4
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    • pp.1857-1862
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    • 2013
  • Dermal fibroblasts produce the many components of the extracellular matrix (ECM) that are needed to maintain connective tissue integrity and repair tissue injuries. This study investigated the effects of Areca catechu extract (ACE) on dermal fibroblast cell activation. Cultured human dermal fibroblasts were treated with ACE, and then ECM production was determined by ELISA, Western blot and RT-PCR. ACE significantly accelerated the production of type 1 collagen, fibronectin, and transforming growth factor (TGF)-${\beta}1$ by ELISA and type 1 collagen by Western blot assay. ACE also increased the gene expression of COL1A1, TGF-${\beta}1$, keratinocyte growth factor (KGF) and insulin growth factor (IGF)-1. These results suggest that ACE has the potential to stimulate ECM production and that it might be suitable for maintaining skin texture.

Optimal Condition for Mycelial Growth of Beauveria bassiana and Its Extracellular Enzyme Activity (백강균(Beauveria bassiana)의 균사체 최적 배양조건 및 효소활성)

  • 민응기;한영환
    • Korean Journal of Microbiology
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    • v.38 no.1
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    • pp.50-53
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    • 2002
  • The optimum temperature and pH for mycelial growth of B. bassiana DGUM 34001 were $24^{\circ}C$ and pH 7.0, respectively. Among the complex media used, mushroom complex medium (MCM) was the most favorable for mycelial growth. When Czapek-Dox medium was used as a minimal medium, glucose was an excellent source for carbon and energy. Soytone and sodium phosphate were favorable constituent for culture medium as a source of organic nitrogen and phosphorus, respectively. When the fungus was grown in MCM broth, the specific activity of extracellular enzyme of ${\alpha}$-amylase, lipase, chitinase, CMCase and pretease were 297.0, 0.058, 0.33, 0.21 and 22.8 units/mg protein, respectively. When various sources of organic nitrogen and chitin were supplemented to determine the production of enzymes, casein and soluble chitosan enhanced the production of extracellular protease and chitinase.

Secretion of the iron containing superoxide dismutase of Streptomyces subrutilus P5 (Streptomyces subrutilus P5가 생산하는 철 함유 superoxide dismutase의 분비)

  • Park, Jae-seung;Kim, Jae-heon
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.108-114
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    • 2015
  • We tried to analyze the growth time for secretion of the iron containing superoxide dismutase by comparing the intra-and extracellular enzyme activity from Streptomyces subrutilus P5 and analyze possible genetic information for this enzyme secretion. The mycelial dry weights and glucose concentrations in culture filtrates were determined during growth. Glucose was consumed rapidly during logarithmic growth phase and almost exhausted at 24 h of cultivation. While the intracellular activity of iron containing superoxide dismutase was first appeared at three hours, the extracellular activity of this enzyme appeared from 7.5 h of cultivation, early logarithmic growth phase. This early presence of the superoxide dismutase might not be the result of cell lysis but active secretion pathway. There was no information for signal peptide responsible for the enzyme secretion in sodF. However, we found a type three secretion box in the promoter region of sodF that has been known for the genes of type III secreted proteins in other bacteria. This is the first report on the possible existence of type III secretion in Streptomyces.

Isolation and Charcterization of Micrococcus sp. HJ-19 Secreting Extracellular Protease (단백질 분해효소를 분비하는 Micrococcus sp. HJ19의 분리 및 특성)

  • Cha, In-Tae;Oh, Yong-Sik;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.222-226
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    • 2007
  • Proteases are degradative enzymes which hydrolyze a peptide bond between amino acids and they are abundantly applied to commercial field. In order to screen new source of pretense, bacteria secreting extracellular pretense were isolated by enrichment culture from deep sea water samples of East Sea, Korea. A bacterium, named as HJ19, showed the best growth and the largest clear zone in plates supplemented skim milk at $30^{\circ}C$. The partial DNA sequence analysis of the 16S rRNA gene, phenotypic tests and morphology identified that this strain was In genus Micrococcus. The strain HJ19 could not grow at $10^{\circ}C$ but it started growth and showed pretense activity at $20^{\circ}C$. The optimal growth was at $37^{\circ}C$ and the maximal protease activity at $30^{\circ}C$ was about 480unit/ml.

Study on the Optimum Cultural Conditions for the production of Extracellular Polysaccharide by Lactic Acid Bacteria (유산균이 생성하는 세포외 다당류의 최적 생성조건에 관한 연구)

  • 강국희;정후길
    • Journal of Food Hygiene and Safety
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    • v.5 no.1
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    • pp.27-34
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    • 1990
  • As a link in the studies on the extracellular polysaccharide by lactic acid bacteria, the experiment was conducted to investigate the viscosity variations of 10% reconstituted skim milk and optimum cultural conditions for the production of extracellular polysaccharide. 1. The viscosity of 10% reconstituted skim milk cultured by 8tr. thermophilu8 510 reached 5,000 CP at $41^{\circ}C$ and on 5 days. Because it was the highest value among the microorganisms tested, Str. thermophilu8 510 was chosen as a main test microoganism and studied on factors affecting maximum viscosity depending upon production of exopolysaccharide. 2. Absorbance of synthetic culture medium was highest at $41^{\circ}C$. Sucrose, in a member of carbohydrates, was the most effective carbon source for the production of exopolysaccharide within 5% concentration. 3. The production of exopplysaccharide was stimulated by $Mg^{++}$.

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Antiapoptotic Effect of Aurintricarboxylic Acid; Extracellular Action versus Inhibition of Cytosolic Protein Tyrosine Phosphatases

  • Lee, Dong-Yoon;Kim, Mee-Kyung;Kim, Mi-Jeong;Bhattarai, Bharatraj;Kafle, Bhooshan;Lee, Keun-Hyeung;Kang, Jae-Seung;Cho, Hyeong-Jin
    • Bulletin of the Korean Chemical Society
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    • v.29 no.2
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    • pp.342-346
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    • 2008
  • Aurintricarboxylic acid (ATA) prevents apoptosis in a wide range of cell types, including PC12 cells. ATA is known to increase the phosphorylation level of IGF-1 receptor (IGF-1R) and downstream signaling proteins. ATA can translocate across the plasma membrane of PC12 cells and inhibit protein tyrosine phosphatases (PTPs) and, therefore, it is not clear whether ATA exerted its antiapoptotic effect through activation of IGF-1R or by inhibition of cytosolic PTPs. When PC12 cells, deprived of serum, were treated with Fab fragment of anti-IGF-1R antibody to prevent the binding of ATA to the extracellular domain of IGF-1R, ATA was found to penetrate into the cytosolic space of the cells. Under these conditions, the survival-promoting effects of ATA were abolished, and the increase of phosphorylation and characteristic cleavage of IGF-1R were not observed. These results indicate that the antiapoptotic effect of ATA in PC12 cells is due to the binding of ATA to the extracellular domain of IGF-1R and subsequent activation of the IGF-1R, not inhibition of cytosolic PTP(s).