• Title/Summary/Keyword: vesicle

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Characterization of ATPase Activity of Free and Immobilized Chromatophore Membrane Vesicles of Rhodobacter sphaeroides

  • Kim, Hyeonjun;Tong, Xiaomeng;Choi, Sungyoung;Lee, Jeong K.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.12
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    • pp.2173-2179
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    • 2017
  • The intracytoplasmic membrane of Rhodobacter sphaeroides readily vesiculates when cells are lysed. The resulting chromatophore membrane vesicle (CMV) contains the photosynthetic machineries to synthesize ATP by ATPase. The light-dependent ATPase activity of CMV was lowered in the presence of $O_2$, but the activity increased to the level observed under anaerobic condition when the reaction mixture was supplemented with ascorbic acid (${\geq}0.5mM$). Cell lysis in the presence of biotinyl cap phospholipid (bcp) resulted in the incorporation of bcp into the membrane to form biotinylated CMV (bCMV), which binds to streptavidin resin at a ratio of approximately $24{\mu}g$ bacteriochlorophyll a/ml resin. The ATPase activity of CMV was not affected by biotinylation, but approximately 30% of the activity was lost by immobilization to resin. Interestingly, the remaining 70% of ATPase activity stayed constant during 7-day storage at $4^{\circ}C$. On the contrary, the ATPase activity of bCMV without immobilization gradually decreased to approximately 40% of the initial level in the same comparison. Thus, the ATPase activity of CMV is sustainable after immobilization, and the immobilized bCMV can be used repeatedly as an ATP generator.

Altered Complexin Expression in Psychiatric and Neurological Disorders: Cause or Consequence?

  • Brose, Nils
    • Molecules and Cells
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    • v.25 no.1
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    • pp.7-19
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    • 2008
  • Complexins play a critical role in the control of fast synchronous neurotransmitter release. They operate by binding to trimeric SNARE complexes consisting of the vesicle protein Synaptobrevin and the plasma membrane proteins Syntaxin and SNAP-25, which are key executors of membrane fusion reactions. SNARE complex binding by Complexins is thought to stabilize and clamp the SNARE complex in a highly fusogenic state, thereby providing a pool of readily releasable synaptic vesicles that can be released quickly and synchronously in response to an action potential and the concomitant increase in intra-synaptic $Ca^{2+}$ levels. Genetic elimination of Complexins from mammalian neurons causes a strong reduction in evoked neurotransmitter release, and altered Complexin expression levels with consequent deficits in synaptic transmission were suggested to contribute to the etiology or pathogenesis of schizophrenia, Huntington's disease, depression, bipolar disorder, Parkinson's disease, Alzheimer's disease, traumatic brain injury, Wernicke's encephalopathy, and fetal alcohol syndrome. In the present review I provide a summary of available data on the role of altered Complexin expression in brain diseases. On aggregate, the available information indicates that altered Complexin expression levels are unlikely to have a causal role in the etiology of the disorders that they have been implicated in, but that they may contribute to the corresponding symptoms.

Oocytes Maturation by Follicular Culture in Porcine (돼지난포의 배양에 의한 난자의 성숙)

  • 박춘근;임종민;박영국;이준희;이상영;정희태;양부근;김정익
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.117-122
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    • 1997
  • The aim of this study was to investigate the effect of the follicular culture from which the oocytes originate on their subsequent in vitro maturation ability. Ovarian follicles were isolated and cultured according to size(1~2mm, 2~6mm and 6~8mm) for 42~44 h. The rates of germinal vesicle breakdown(GVBD) in each groups were 87%(65/75), 82%(80/97) and 89%(47/53), but the oocytes maturation were su, pp.essed at anaphase-I stage. In spite of the adding porcine follicular fluid and/or hormones in maturation medium, maturation ability of oocytes from follicle cultured for 21~22 h were inhibited. When oocytes from follicle cultured for 4 h at various temperature were incubated for 38~40 h, the rates of oocytes maturation from follicle cultured at 2$0^{\circ}C$(51%, 26/51) and 39$^{\circ}C$(54%, 26/48) were significant higher(P<0.05) than group cultured at 4$^{\circ}C$(33%, 19/58). On the other hand, the GVBD were stared 2 h after culture of follicle of oocytes. To summairze, oocytes maturation by follicular culture were inhibited at anaphase-I stage in porcine. When the follicle cultured for 4 h, maturation were completed to metaphase-II stage. However, rates of GVBD in oocytes from follicular culture were higher than oocytes cultured in medium.

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The Functional Role of Maturation Promoting Factor in the Two-cell Embryos (생쥐 2-세포기 배아에서 성숙유도물질의 기능적 역할)

  • 강해묵;이대기
    • The Korean Journal of Zoology
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    • v.36 no.2
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    • pp.277-284
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    • 1993
  • A possible involvement of maturation promoting factor (nfPF) in the two-cell block phenomenon was studied by fusion experiments. Germinal vesicle (GlF) ooeyte was fused with a blastomore from late or blocked 2-cell mouse embryos. and germinal vesicle breakdoum (GVBD) of fused GV oocvtes in the presence of dbcAMP (100$\mu$g/ml) was scored as an index of MPF aniviD. GnD was induced approximately 30% by fusion of a blastomere derived from late 2-cell embryos, but not from blocked 2-cell embryos. The rate of GVBD was changed when GV oocyte was fused with a blastomere from late 2-cell embryos which were treated with u-amanitin, puromvcin or colcemid before and after hsion: Treatment of late 2-cell embryos with puromycin (50 Is/mll but not with u-amanitin (100 Is/ml) clearly inhibited GVBD, indicating that do novo protein synthesis maw be required for the appearance of MPF activity in late 2-cell embryos. Treatment of late 2-cell embryos w기h colcemid (0.1 Is/mll doubled GVBD, presumably due to the maintenance of metaphase or mitotic phase. SDS-PAGE and twoiimensional electrophoresis revealed that there was no difference in protein synthetic pattern in late and blocked 2-cell embryos, but three phosphoproteins with 27, 35 and 46 M)a, presumsblv M-phase components were phosphorylated in late 2-cell embryos but not in blocked 2-cell embryos. It seems then that MPF activity is closely related to phosphorylstion of M-phase components in late 2-cell embryos.

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The Effect of Laser Irridation on the Ultrastructure of Retina (Laser 조사가 망막의 미세구조에 미치는 영향)

  • Kim, Douk Hoon;Mun, Jung Hak
    • Journal of Korean Ophthalmic Optics Society
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    • v.1 no.1
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    • pp.15-22
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    • 1996
  • The fine structure of retinal tissue was studied to investigate on effect of Laser irridation on the ICR mouse with electron microscope. The results obtained were as follows: 1. At the normal groups, the most retinal layers were a complex structure, consisting of several specific cells and nerve fiver. 2. In the increasing time of Laser irridation, each cell layer of retina was not uniform of the structure and band. The visual cells were severely heterochromatin swelling of cytoplasm, irregular shape & heterochromatin of nuclear, and disappear of some cytoplasm. The nucleus and nerve fiber of retinal layer was a very irregular shape, formation of vesicle, not identify of each intercellular boundary. The pigment epithelial cells were not an uniform, a large vesicle formation of cytoplasm, and a condensation & very irregular shape of nucleus.

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Affinity between TBC1D4 (AS160) phosphotyrosine-binding domain and insulin-regulated aminopeptidase cytoplasmic domain measured by isothermal titration calorimetry

  • Park, Sang-Youn;Kim, Keon-Young;Kim, Sun-Min;Yu, Young-Seok
    • BMB Reports
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    • v.45 no.6
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    • pp.360-364
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    • 2012
  • Uptake of circulating glucose into the cells happens via the insulin-mediated signalling pathway, which translocates the glucose transporter 4 (GLUT4) vesicles from the intracellular compartment to the plasma membrane. Rab GTPases are involved in this vesicle trafficking, where Rab GTPases-activating proteins (RabGAP) enhance the GTP to GDP hydrolysis. TBC1D4 (AS160) and TBC1D1 are functional RabGAPs in the adipocytes and the skeletonal myocytes, respectively. These proteins contain two phosphotyrosine-binding domains (PTBs) at the amino-terminus of the catalytic RabGAP domain. The second PTB has been shown to interact with the cytoplasmic region of the insulin-regulated aminopeptidase (IRAP) of the GLUT4 vesicle. In this study, we quantitatively measured the ${\sim}{\mu}M$ affinity ($K_D$) between TBC1D4 PTB and IRAP using isothermal titration calorimetry, and further showed that IRAP residues 1-49 are the major region mediating this interaction. We also demonstrated that the IRAP residues 1-15 are necessary but not sufficient for the PTB interaction.

Mixed infection of bovine papillomavirus and parapoxvirus in dairy cattle with vesidle-like lesions (유우에서 발생한 수포양 병변에서 bovine papillomavirus 와 parapoxvirus 혼합감염)

  • Lyoo, Young S;Kang, Yung-bai;Chang, Chung-ho;Moon, Oun-gyong;Choi, Sang-ho;Park, Joong-won
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.353-358
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    • 1998
  • A mysterious disease with vesicle-like nodules in dairy cattle udder has been drawn attention to the foreign animal disease expert at the National Veterinary Research Institute. Immediately dispatched foreign animal disease expert, pathologist and regional veterinary officials executed quarantine nearby affected farm area to prevent transmission of the pathogen which was possibly contagious vesicular disease agents in domestic animals such as FMD or SVD. Proper samples were collected for the laboratory examination. Vesicle-like lesions were only detected in lactating dairy cattle and no distinct clinical signs have been observed in affected animals. Parapox and papillomavirus particles have been demonstrated on electromicroscopical examination from nodular samples of udder lesions of the dairy cattle. Characteristic papilloma virus particles with 55nm in diameter and parapoxvirus with 150nm in diameter and 350nm long oval shape particles were detected and confirmed by embryonated chicken egg inoculation.

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Effect of Trehalose on Biological Membranes with Respect to Phase of the Membranes

  • Park, Jin-Won
    • KSBB Journal
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    • v.32 no.2
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    • pp.103-107
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    • 2017
  • The effect of the trehalose incorporation on the biological membranes was investigated with respect to the phase of the membranes using the fluorescence intensity change. Spherical phospholipid bilayers, vesicles, were prepared only with the variation in the phase of each layer via a double emulsion technique. In the aqueous inside of the vesicles, 8-Aminonaphthalene-1,3,6-trisulfonic acid disodium salt(ANTS) was encapsulated. As a quencher, p-Xylene-bis(N-pyridinium bromide)(DPX) was included in the buffer where the vesicles were dispersed. The fluorescence scale was calibrated with the fluorescence of ANTS vesicles in p-Xylene-bis(N-pyridinium bromide)(DPX)-included-buffer taken as 100% fluorescence and the mixture of ANTS and DPX in the buffer as 0% fluorescence. Trehalose injection into the vesicle solution led the distortion of the membrane. It was found that the distortion was related to the phase of each layer the vesicle up on the ratio of trehalose to lipid. In the identical measurements at glucose, the behavior of the distortion was completely different from that of trehalose. These results seem to depend on the stability of the vesicles, due to the osmotic and volumetric effects on the headgroup packing disruption.

Reduction of Twin Pregnancy by Transvaginal Ultrasound-guided Aspiration in a Mare

  • Lee, Eun-bee;Song, Mingeun;Park, Chull-gyu;Hwang, Jun-seok;Chun, Yong-woo;Lee, Seung-hwan;Cheong, Jongtae;Lee, Joomyoung;Kang, Tae-Young;Seo, Jong-pil
    • Journal of Veterinary Clinics
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    • v.34 no.4
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    • pp.304-306
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    • 2017
  • A 3-year-old Thoroughbred mare was referred to J&C Equine Hospital with gestation day 30 twin pregnancy. On transrectal ultrasonographic examination, two similar sized (28 mm) embryonic vesicles, unilaterally fixed in the uterine horn, were detected. Transvaginal ultrasound-guided aspiration (TUGA) was performed for reduction of one embryonic vesicle. Yolk sac and allantoic cavity fluids from one embryonic vesicle were aspirated by a needle guided by using transvaginal ultrasound. The mare continued normal singleton pregnancy after twin reduction and delivered a foal successfully. This is the first case described the clinical use of TUGA in Korea. Clinical use of TUGA in twin reduction after embryonic fixation is recommended for equine clinicians.

Effect of Co-Culture with Mammalian Spermatozoa on In Vitro Maturation of Porcine Cumulus-Enclosed Germinal Vesicle Oocytes

  • Kim Byung Ki
    • Reproductive and Developmental Biology
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    • v.28 no.4
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    • pp.235-240
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    • 2004
  • In vitro maturation of denuded porcine immature oocytes can be enhanced by co-incubation with spermatozoa even before fertilization. This study was to determine whether the addition of spermatozoa into the culture medium could influence the nuclear maturation of porcine cumulus-enclosed germinal vesicle (GV) oocytes. Cumulus-oocyte complexes (COCs) were collected from follicles of 3- to 5-mm diameter. Porcine COCs were cultured in tissue culture medium containing spermatozoa. After 48 h culture, oocytes were examined for evidence of GV breakdown, metaphase I, anaphase-telophase I, and metaphase II. The proportion of oocytes reaching at metaphase II was significantly (P < 0.05) increased in the oocytes cultured in media containing spermatozoa compared to those in media without spermatozoa (52.3% vs 12.5%). No difference in the percentage of metaphase II was observed among the different periods of spermatozoa exposure and among the spermatozoa from different species. The proportion of oocytes reaching metaphase II was significantly different between high and low concentrations of spermatozoa. The present study suggests that manunalian spermatozoa contain a substance(s) that improves nuclear in vitro maturation of porcine cumulus-enclosed GV oocytes. Enhancing effect of spermatozoa for in vitro maturation of oocytes is a highly dose-dependent.