• 제목/요약/키워드: intact cells

검색결과 444건 처리시간 0.029초

Correlation Between Electrical Activity of Type I Neuron and c-Fos Expression in the Medial Vestibular Nuclei Following Unilateral Labyrinthectomy in Rats

  • Park, Byung-Rim;Doh, Nam-Yong;Kim, Min-Sun;Chun, Sang-Woo;Lee, Moon-Young;Lee, Sung-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • 제1권5호
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    • pp.505-513
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    • 1997
  • To search the correlations between electrical activity and c-Fos expression in the process of vestibular compensation, we examined the changes of those two parameters in the medial vestibular nuclei (MVN) of unilaterally labyrinthectomized (ULX) rats. Spontaneous nystagmus with fast component toward the intact side disappeared gradually within 48 hours. Fourty eight hours after ULX, directional preponderance of the eye movement induced by sinusoidal rotation of the whole body which represents the symmetry of bilateral vestibular functions showed less than 20% by rotation of 0.1, 0.2, and 0.5 Hz, indicating the recovery of symmetry in bilateral vestibular functions. Six hours after ULX, spontaneous electrical activity of type I neurons resulted in asymmetry between bilateral MVN, however, the asymmetry of the electrical activity was decreased 48 hours after ULX. Immunocytochemical staining revealed that ULX produced dramatic induction of c-Fos positive cells in the MVN bilaterally. The number of c-Fos immunoreactive cells in the contralateral MVN was significantly higher than those in the ipsilateral MVN (p<0.0001) 2 hours after ULX. Thereafter, the number of c-Fos positive cells decreased bilaterally and was slightly, but not significantly higher in the ipsilateral MVN at 48 hours after ULX. The present results suggest that both electrical activity of type I neurons and c-Fos expression in MVN following ULX will reflect underlying mechanisms of recovery process of vestibular compensation.

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한우에서 거세에 의한 Testosterone 결핍이 사이토카인의 변화에 미치는 영향 연구 (The change aspects between testosterone and cytokines by castration in Hanwoo cattle)

  • 김성범;조아라;최창용;정영훈;류재규;김은주;도윤정
    • 한국동물위생학회지
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    • 제40권4호
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    • pp.227-233
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    • 2017
  • The purpose of this study is to determine the changes of cytokines and immune cells in blood in castrated Hanwoo. The cytokine production and the number of immune cells were determined by collecting blood from jugular vein before castration and on 1, 7, and 28 days then after. Results of the hematological test showed that the number of leukocyte tend to increase after castration, and it significantly decreased on day 7 and day 28 (P<0.05). Lymphocytes decreased significantly on day 1 and 7 (P<0.05), and then recovered as in neutrophils on day 28. The levels of serum testosterone, TNF-a, IL-6, and anti-inflammatory IL-10 significantly decreased (P<0.05) after castration. There was also a decrease in IL-2, $IFN-{\gamma}$, and IL-4 but showed no significant difference when compared to intact ones. These results suggest that testosterone-deficiency does not affect the number of immune cells in blood, but has a close relationship with cytokine production.

내피세포성 이완인자에 대한 세포외 $Ca^{2+}$$Ca^{2+}$-길항제의 작용기전 (Acting Mechanisms of Extracellular$Ca^{2+}$ and $Ca^{2+}$ - antagonists on Endothelium - Derived Relaxing Factor in Rabbit Aorta.)

  • 진성훈
    • Journal of Chest Surgery
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    • 제24권3호
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    • pp.229-244
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    • 1991
  • A bioassay technique and organ bath study were performed to analyze the effects of extracellular $Ca^{2+}$ and $Ca^{2+}$-antagonists on endothelium-derived relaxing factor[s][EDRF] released from the endothelial cells of rabbit aorta. Transverse strips with intact endothelium or damaged endothelium were used for the mechanical contraction experiment using organ bath. Long segment including thoracic and abdominal aorta with endothelium [EDRF donor aorta] was perfused with Tyrode solution which was aerated with 95% $O_2-5%$ $CO_2$ mixed gas and kept at 35oC. The perfusate was bioassayed with a transverse strip of thoracic aorta with damaged endothelium. The test strip was contracted with nor-epinephrine and acetylcholine was used to stimulate the release of EDRF from endothelial cells. The results obtained were as follows; 1] The endothelium-dependent relaxation[EDR] induced by acetylcholine was biphasic; an initial rapid relaxation followed by a slow relaxation. 2] EDR induced by acetylcholine was reduced gradually with the decrease in the concentration of extracellular $Ca^{2+}$. The effect of extracellular $Ca^{2+}$ on EDR was more prominent in the late slow relaxation phase. 3] EDR to acetylcholine was not altered by acute exposure to organic $Ca^{2+}$-antagonists. Pretreatment with verapamil to the EDRF donor aortic segment did not alter the magnitude of EDR. 4] Among the inorganic $Ca^{2+}$-antagonists $Mn^{2+}$ and $Cd^{2+}$ did not inhibit EDR, whereas $Co^{2+}$ and $La^{3+}$ inhibited EDR. 5] The inhibitory response of $Co^{2+}$ to EDR developed when infused directly on the test strip. That of $La^{3+}$, however, was evoked when added to solution perfusing the donor aortic segment. The above results suggest that $Ca^{2+}$-antagonists do not affect EDR and the inhibitory effect of $Ca^{2+}$ results from influencing the action of EDRF on vascular smooth muscle, whereas that of $La^{3+}$ results from its action on the release of EDRF from endothelial cells.

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Time-Lapse Video Microscopy of Wound Recovery and Reproduction in the Siphonous Green Alga Derbesia tenuissima

  • ;;김광훈
    • ALGAE
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    • 제21권1호
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    • pp.109-124
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    • 2006
  • Responses to various types of mechanically induced wounding were followed in the giant-celled Caulerpalean species, Derbesia tenuissima, using time-lapse video-microscopy. Gametophyte vesicle cells. Puncture wounding: the gametophyte cell seals the puncture in 5 min. This is followed by cycles of ruptures and sealing, ending with full recovery in 24 hrs. Cut wounding: the protoplast immediately retracts away from the wall and reforms an intact, deflated protoplast that expands to fill the original cell within 21 hrs. Crush wounding (internal). When retained within the cell wall many protoplast fragments condense, round up, and coalesce; the reconstituted protoplast expands until it attains complete recovery, filling the original cell shape in 12 hrs. Crush wounding (external). Protoplast fragments extruded from the crushed cell are more numerous and smaller taking longer to recover. Most fragments become spherical, transforming into small viable cells capable of reproduction in several days. Sporophyte filaments. Crush wounding creates many small fragments that initially condense, coalesce and then expand within the wall to restore a complete filament with normal cytoplasmic streaming within 5 hrs. Reproduction: gametophyte. Our culture isolates produce more females than males (30:1). Gametangia develop one day before discharge that occurs explosively (1/6 sec) at first morning light. The vesicle cell forms successive gametangia every 14 days. Sporophyte. Each sporangium develops on a lateral branch that becomes isolated by the creation of successive basal plugs. After cytoplasmic cleavage and differentiation the stephanokont spores are discharged. The spores settle quickly and germinate forming gametophyte cells.

고구마 배양세포에서 Peroxiredoxin cDNA의 분리 및 발현 특성 (Molecular Cloning and Characterization of a Peroxiredoxin cDNA from Cell Cultures of Sweetpotato)

  • 박수영;류선화;권석윤;김종국;곽상수
    • Journal of Plant Biotechnology
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    • 제30권2호
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    • pp.135-141
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    • 2003
  • Peroxiredoxin(Pix) are large family of peroxidases that reduce alkyl hydroperoxides and hydrogen peroxide. A cDNA clone (referred to as swPrxl) encoding Pix was from a sweetpotato cDNA library constructed from suspension-sultured cells, and its expression was investigated in terms of stress. The swPrxl contained an open reading frame (ORF) encoding mature protein of 193 amino acids with calculated molecular mass of 20.8kDa. The predicted amino acid sequence of swPrxl has two conserved cysteines that are essential resicues for the reduction of peroxides. It showed high amino acid sequence homology ot PixIIF of Arabidopsis (77%) and putative Prx of rice(72%). RNA gel-blot analysis showed that swPrxl gene was expressed dominantly in leave among intact tissues, and also highly detect in suspension-cultured cells. Interestingly, the level of swPrxl transcripts was almost the same regardless of the growth stage in suspension culture. Furthermore, the transcription level of swPrxl gene was not significantly changed in response to various stress treatments such as wounding, extreme temperature and stress-related chemicals RT-PCR analyses.

Ultrastructures of the Loaves of Cucumber Plane Treated with DL-3-Aminobutyric Acid at the Vascular Bundle and the Penetration Sites after Inoculation with Colletotrichum orbiculare

  • Jeun, Y.C.;Park, E.W.
    • The Plant Pathology Journal
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    • 제19권2호
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    • pp.85-91
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    • 2003
  • Pre-treatment with DL-3-aminobutyric acid (BABA) in the cucumber plants caused the decrease of disease severity after inoculation with anthracnose pathogen Colletotrichum orbiculare. In this study, ultrastructures of the vascular bundle and the infection structures in the leaves of BABA-treated as well as untreated cucumber plants were observed after inoculation with the anthracnose pathogen by electron microscopy. The ultrastructures of vascular bundle in the leaves of BABA-treated plants were similar to those of the untreated plants except plasmodesmata. In the BABA-treated plants, the plasmodesmata were more numerous than in the untreated plants, suggesting that the BABA treatment may cause the active transfer of metabolites through the vascular bundle. In the leaves of untreated plants, the fungal hyphae were spread widely in the plant tissues at 5 days after pathogen inoculation. Most cellular organelles in the hyphae were intact, indicating a compatible interaction between the plant and the parasite. In contrast, in the leaves of BABA pre-treated plants the growth of most hyphae was restricted to the epidermal cell layer at 5 days after inoculation. Most hyphae cytoplasm and nucleoplasm was electron dense or the intracellular organelles were degenerated. The cell walls of some plant cells became thick at the site adjacent to the intercellular hyphae, indicating a mechanical defense reaction of the plant cells against the fungal attack. Furthermore, hypersensitive reaction (HR) of the epidermal cells was often observed, in which the intracellular hyphae were degenerated. Based on these results it is suggested that BABA causes the enhancement of defense mechanisms in the cucumber plants such as cell wall apposition or HR against the invasion of C. orbiculare.

Expression and Activation of Transforming Growth Factor-Beta 2 in Cultured Bone Cells

  • Lee, Chang-Ho
    • Animal cells and systems
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    • 제4권3호
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    • pp.273-278
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    • 2000
  • Transforming growth factor-$\beta$ (IGF-$\beta$)s are multifunctional small polypeptides synthesized in most cell types. TGF-$\beta$ exerts pivotal effects on both bone formation and resorption. In addition, increasing lines of evidence implicate TGF-$\beta$ as a potential coupling factor between these two processes during bone remodeling. In the present study, the expression form and the activation mechanism of latent-TGF-$\beta$ were investigated using specific antibodies for each isoform. TGF-$\beta$s were observed to be synthesized and accumulated in a large amount in cultured osteoblastic cells. The estimated molecular weights of intracellular TGF-$\beta$2 and -$\beta$3 were 49 and 55 kDa, respectively. Based on proteolytic digestion study and immunofluorescence observation, these precursor forms seemed to be accumulated in distinct intracellular compartments. To examine whether the internal pool of TGF-$\beta$ was possiblely regulated by external signals, their biological activites were examined in a conditioned media of this cell. Although the intact conditioned media did not contain detectable TGF-$\beta$ activity, heat-treatment or acid-activation of the conditioned media revealed significant TGF-$\beta$ activity. Furthermore, in the presence of estrogen, this activity was dramatically diminished. It is known that activation of latent TGF-$\beta$ can be achieved by different chemical and enzymatic treatments, or by incubation with certain cell types. This extracellular activation was suggested as a key step in the regulation of TGF-$\beta$ activity. In addition to these extracellular activation, this study suggests that the synthesis and intracellular processing are important regulation steps for TGF-$\beta$ action. In addition, this regulation Is specific for TGF-$\beta$ type 2, because the change was not observed in TGF-$\beta$3 in osteoblastic cell line.

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Inhibitory Effects of Verapamil and TMB-8 on Tonic Contraction Are Accompanied by Inhibition of Phospholipase C Activity in Intact Gastric Smooth Muscle Cells

  • Sim, Sang-Soo;Yoon, Shin-Hee;Hahn, Sang-June;Rhie, Duck-Joo;Jo, Yang-Hyeok;Kim, Myung-Suk
    • The Korean Journal of Physiology
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    • 제29권1호
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    • pp.29-37
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    • 1995
  • Gastric smooth muscle of guinea pigs was used to investigate whether the inhibitory effect of calcium antagonists on tonic contraction was accompanied by inhibition of phospholipase C activity. Tonic contraction and $[^{3}H]$ inositol phosphate (IP) formation in response to acetylcholine were measured after pretreatment with verapamil, nifedipine, 8-(N,N-diethylamino)octyl 3,4,5-trimethoxy-benzoate (TMB-8) or EGTA. Verapamil $(10\;{\mu}M)$, TMB-8 $(10\;{\mu}M)$ or EGTA (2 mM) significantly inhibited acetylcholine $(1\;{\mu}M)$-stimulated tonic contraction but nifedipine (100 nM) did not. Acetylcholine dose-dependently increased the formation of $[^{3}H]IP$. This effect was not observed in the presence of 2 mM EGTA. Both verapamil and TMB-8 significantly inhibited $[^{3}H]IP$ formation induced by $10\;{\mu}M$ acetylcholine, whereas nifedipine did not. In a subsequent study, we measured phospholipase C activity in gastric muscle cell homogenate and in permeabilized cells to determine whether calcium antagonists could inhibit the activity directly. The calcium antagonists did not change the phospholipase C activity of the cell homogenate or the permeabilized cells. But EGTA decreased phospholipase C activity by 50%. These results suggest that the inhibitory effects of verapamil and TMB-8 on acetylcholine-stimulated tonic contraction may be accompanied by inhibition of phospholipase C activity.

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Nanoscale imaging of rat atrial myocytes by scanning ion conductance microscopy reveals heterogeneity of T-tubule openings and ultrastructure of the cell membrane

  • Park, Sun Hwa;Kim, Ami;An, Jieun;Cho, Hyun Sung;Kang, Tong Mook
    • The Korean Journal of Physiology and Pharmacology
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    • 제24권6호
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    • pp.529-543
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    • 2020
  • In contrast to ventricular myocytes, the structural and functional importance of atrial transverse tubules (T-tubules) is not fully understood. Therefore, we investigated the ultrastructure of T-tubules of living rat atrial myocytes in comparison with ventricular myocytes. Nanoscale cell surface imaging by scanning ion conductance microscopy (SICM) was accompanied by confocal imaging of intracellular T-tubule network, and the effect of removal of T-tubules on atrial excitation-contraction coupling (EC-coupling) was observed. By SICM imaging, we classified atrial cell surface into 4 subtypes. About 38% of atrial myocytes had smooth cell surface with no clear T-tubule openings and intracellular T-tubules (smooth-type). In 33% of cells, we found a novel membrane nanostructure running in the direction of cell length and named it 'longitudinal fissures' (LFs-type). Interestingly, T-tubule openings were often found inside the LFs. About 17% of atrial cells resembled ventricular myocytes, but they had smaller T-tubule openings and a lower Z-groove ratio than the ventricle (ventricular-type). The remaining 12% of cells showed a mixed structure of each subtype (mixed-type). The LFs-, ventricular-, and mixed-type had an appreciable amount of reticular form of intracellular T-tubules. Formamide-induced detubulation effectively removed atrial T-tubules, which was confirmed by both confocal images and decreased cell capacitance. However, the LFs remained intact after detubulation. Detubulation reduced action potential duration and L-type Ca2+ channel (LTCC) density, and prolonged relaxation time of the myocytes. Taken together, we observed heterogeneity of rat atrial T-tubules and membranous ultrastructure, and the alteration of atrial EC-coupling by disruption of T-tubules.

Ginsenosides repair UVB-induced skin barrier damage in BALB/c hairless mice and HaCaT keratinocytes

  • Li, Zhenzhuo;Jiang, Rui;Wang, Manying;Zhai, Lu;Liu, Jianzeng;Xu, Xiaohao;Sun, Liwei;Zhao, Daqing
    • Journal of Ginseng Research
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    • 제46권1호
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    • pp.115-125
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    • 2022
  • Background: Ginsenosides (GS) have potential value as cosmetic additives for prevention of skin photoaging. However, their protective mechanisms against skin barrier damage and their active monomeric constituents are unknown. Methods: GS monomer types and their relative proportions were identified. A UVB-irradiated BALB/c hairless mouse model was used to assess protective effects of GS components on skin epidermal thickness and transepidermal water loss (TEWL). Skin barrier function, reflected by filaggrin (FLG), involucrin (IVL), claudin-1 (Cldn-1), and aquaporin 3 (AQP3) levels and MAPK phosphorylation patterns, were analyzed in UVB-irradiated hairless mice or HaCaT cells. Results: Total GS monomeric content detected by UPLC was 85.45% and was largely attributed to 17 main monomers that included Re (16.73%), Rd (13.36%), and Rg1 (13.38%). In hairless mice, GS ameliorated UVB-induced epidermal barrier dysfunction manifesting as increased epidermal thickness, increased TEWL, and decreased stratum corneum water content without weight change. Furthermore, GS treatment of UVB-irradiated mice restored protein expression levels and epidermal tissue distributions of FLG, IVL, Cldn-1, and AQP3, with consistent mRNA and protein expression results obtained in UVB-irradiated HaCaT cells (except for unchanging Cldn-1 expression). Mechanistically, GS inhibited JNK, p38, and ERK phosphorylation in UVB-irradiated HaCaT cells, with a mixture of Rg2, Rg3, Rk3, F2, Rd, and Rb3 providing the same protective MAPK pathway inhibition-associated upregulation of IVL and AQP3 expression as provided by intact GS treatment. Conclusion: GS protection against UVB-irradiated skin barrier damage depends on activities of six ginsenoside monomeric constituents that inhibit the MAPK signaling pathway.