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Effects of Chlorpromazine·HCl on the Structural Parameters of Bovine Brain Membranes

  • Jang, Hye-Ock;Jeong, Dong-Keun;Ahn, Shin-Ho;Yoon, Chang-Dae;Jeong, Soo-Cheol;Jin, Seong-Deok;Yun, Il
    • BMB Reports
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    • v.37 no.5
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    • pp.603-611
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    • 2004
  • Fluorescence probes located in different membrane regions were used to evaluate the effects of chlorpromazine HCl on structural parameters (transbilayer lateral mobility, annular lipid fluidity, protein distribution, and lipid bilayer thickness) of synaptosomal plasma membrane vesicles (SPMVs) isolated from bovine cerebral cortex. The experimental procedure was based on the selective quenching of 1,3-di(1-pyrenyl)propane (Py-3-Py) by trinitrophenyl groups, radiationless energy transfer from the tryptophan of membrane proteins to Py-3-Py, and energy transfer from Py-3-Py monomers to 1-anilinonaphthalene-8-sulfonic acid (ANS). In this study, chlorpromazine HCl decreased the lateral mobility of Py-3-Py in a concentration dependent-manner, showed a greater ordering effect on the inner monolayer than on the outer monolayer, decreased annular lipid fluidity in a dose dependent-manner, and contracted the membrane lipid bilayer. Furthermore, the drug was found to have a clustering effect on membrane proteins.

STRUCTURAL PERTURBATIONS INDUCED BY PHOTODYNAMIC ACTION OF PORPHYRIN AGGREGATES ON PLASMA MEMBRANE AND MICROSOMES OF GLIOBLASTOMA CELLS

  • Sreentvasan, Rajesh;Joshi, Preeti G.;Joshi, Nanda B.
    • Journal of Photoscience
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    • v.4 no.2
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    • pp.41-48
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    • 1997
  • The plasma membrane and microsomes, isolated from the cells treated with hematoporphyrm derivative (HpD) for 1 and 24 h, accumulated the aggregated porphyrin. The quantity of aggregated porphyrin was same in the plasma membrane and microsomes after isolating them from cells treated with HpD for 1 h whereas the microsomes accumulated higher quantity of aggregated porphyrin when cells were treated with HpD for 24 h. Photodynamic action of aggregated porphyrin on plasma membrane and microsomes was investigated using lipid specific fluorescent probes: 1,6-diphenyl-1,3,5-hexatrine (DPH) and 1-(4-trimethylammonium), 6-diphenyl-1,3,5-hexatrine(TMA-DPH). The time dependent anisotropy of these probes in the membranes was measured and the decay of anisotropy was analyzed using wobbling in cone model. Upon irradiation both the plasma membrane and the microsomes showed an increase in the limiting anis~)tropy and order parameter and a decrease in the cone angle of the lipid probes. The increase in the limiting anisotropy was pronounced in membranes isolated from the cells treated with HpD for 24 h. Photoinduced change in the limiting anisotropy was dependent on the duration of incubation of cells with HpD before isolating the membranes. In both the membranes. the membrane core was affected more as compared to the outer leaflet. In addition to the structural changes, a decrease in Na+-K+-ATPase and NADPH cyt c reductase activity was also observed upon irradiation of HpD treated cells. Inhibition in NADPH cyt c reductase was more when cells were treated with HpD for 24 h, however, Na+-K+-ATPase activity did not depend on the duration of the treatment of cells with HpD before irradiation. Our results suggest that the extent of photoinduced perturbations in the membranes varies as a function of duration of the treatment of cells with HpD and the membrane core is more susceptible to the photodynamic action of aggregated porphyrin.

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Three-dimensional analysis of the arrangement of microtubules of the outer segment in the ciliary-type photoreceptor cell in the Onchidium dorsal eye

  • Katagiri, Nobuko;Shimatani, Yuichi;Katagiri, Yasuo
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.284-286
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    • 2002
  • The inverted retina of the Onchidium dorsal eye (DE) is composed only of ciliary-type photoreceptor cells (CC's). The outer segment (OS) of the CC is a concentric lamellar structure consisting of many modified ciliary membranes and stains positively with anti-β-tubulin antibody. Near the base of the OS there are about 30 basal bodies each connecting individually to a cilium. The cilia are rod-shaped at the base, progressing upwards to a flattened sheet-like shape with increasing surface area. Three-dimensional analysis on serial sections demonstrates the ladle-shape of a modified cilium. Many modified cilia wrap around each other like the leaves of a cabbage. Nine pairs of microtubules (MT's) are located regularly in a ring at the base of the cilium, gradually losing their regular arrangement towards the periphery, where they separate into two subgroups that are contained within two swollen portions of a modified cilium. Within the CC of the Onchidium DE, MT's in the modified cilium exist as two poles extending longitudinally in a thin expanded ciliary membrane. This arrangement may support the photoreceptive OS and serve to maintain its structural integrity.

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Morphogenesis of the Eye of Brown Croaker (Miichthys miiuy)

  • Park, In-Seok;Seol, Dong-Won;Cho, Sung-Hwoan;Song, Young-Chae;Choi, Hee-Jung;Noh, Choong-Hwan;Myoung, Jung-Goo;Kim, Jong-Man
    • Ocean and Polar Research
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    • v.28 no.3
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    • pp.287-290
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    • 2006
  • Eye growth and lens diameter of brown croaker Miichthys miiuy were positively allometric between hatching and 180 days post-hatch (d.p.h.). Eye growth in relation to head length and head height was nearly isometric. Eyes were formed completely at 14 d.p.h. At this age, the eye has a crystalline lens, an optic nerve fiber layer, a ganglion cell layer, an inner plexiform layer, an inner nuclear layer, an outer plexiform layer, an outer nuclear layer, an outer limiting membrane, and a pigment epithelium. The essential demands that must be met by the retina in this species pertain to light sensitivity and spatial resolution.

Light and electron microscopic morphology of the fertilized egg and fertilized egg envelope of Poropanchax normani, Poeciliidae, Teleostei

  • Dong Heui Kim
    • Applied Microscopy
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    • v.52
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    • pp.6.1-6.5
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    • 2022
  • We examined the morphology of the fertilized egg and the fine structure of fertilized egg envelopes of Poropanchax normani belonging to the family Poeciliidae, also known as Norman's lampeye using light and electron microscopes. The fertilized eggs with narrow perivitelline space were found to be spherical and demersal, additionally containing small oil droplets in the vitelline membrane. Further, a bundle of adhesive filaments was observed to be present on one side of the fertilized egg. These filaments possessed remarkably high elasticity and were approximately 1-3mm in length. The size of the fertilized egg was determined to be about 1.49 ± 0.07mm (n=30). The outer surface appeared smooth, and adhesive filaments originating at different location of the surface of the envelope were found to be distributed around the egg envelope and were joined together to form a single long bundle in scanning electron microscopic observation. A peak-like structure formed of several straight wrinkles was observed around the micropyle. However, the complete structure of the micropyle could not be studied due to the depth at which it was located. Additionally, the total thickness of the egg envelope was ascertained to be approximately12.5-14.5㎛. The egg envelope consisted of two distinct layers, an outer electron dense layer and an inner lamellar layer, further consisting of 10 sublayers of varying thicknesses. Collectively, it was observed that the morphological characteristics of the fertilized egg, fine structures surrounding the micropyle, outer surface, adhesive structure consisting adhesive filaments, and sections of fertilized egg envelope displayed species specificity.

Purification and Comparison of NADH-Cytochrome b5 Reductase from Mitochondrial Outer Membrane of Bovine Heart and Turnip

  • 이재양;김영호;이상직
    • Bulletin of the Korean Chemical Society
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    • v.19 no.2
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    • pp.160-164
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    • 1998
  • The NADH-cytochrome b5 reductase (NCBR), a mitochondrial external electron carrier, was purified from bovine heart and turnip and their properties were examined. The mitochondrial outer membranes separated were subjected to NCBR isolation through DEAE-Cellulose ion exchange, DEAE-Sephadex gel chromatography, and hydroxyapatite adsorption chromatography. These processes yielded the purification folds of 88 and 42 and the recovery percentages of 0.2%, 5.67% for turnip and bovine heart, respectively. The molecular weight of the NCBR from the two sources was estimated to be 35,000 using SDS polyacrylamide gel electrophoresis. The Michaelis constant Km and maximum velocity Vmax were determined by measuring the NADH-ferricyanide redox system as well as the NADPH-ferricyanide redox system. The kinetics showed that both NCBRs had higher affinities for NADH than artificial electron-acceptor substrate ferricyanide. Although NADPH had a lower affinity for the enzymes than NADH, this study showed the 2'-phosphate dinucleotide could be used as a substrate.

Effects of chlorhexidine digluconate on thickness of outer membranes isolated from Cultured Porphyromonas gingivalis

  • Jang, Hye-Ock;Ahn, Ki-Weon;Shin, Sang-Hun;Chung, In-Kyo;Yun, Il
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.93.2-93.2
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    • 2003
  • To get a better insight into the biophysical mechanism of action of chlorhexidine digluconate, we examined the effect of chlorhexidine digluconate on the thickness of outer membranes isolated from cultured Porphyromonas gingivalis using energy transfer between the membrane surface fluorescent probe (l-anilinonaphthalene-8-sulfonic acid) and the hydrophobic fluorescent probe [1,3-di(l-pyrenyl)propane]. 1-Anilinonaphthalene-8-sulfonic acid quenches the monomer fluorescence of 1,3-di(1-pyrenyl)propane. (omitted)

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DEVELOPMENT OF A BIOACTIVE CELLULOSE MEMBRANE FROM SEA SQUIRT SKIN FOR BONE REGENERATION - A PRELIMINARY RESEARCH (멍게와 미더덕 피부의 천연 셀룰로오스 각질을 이용한 골재생 효능을 가진 생활성막의 개발 - 예비 연구)

  • Kim, Soung-Min;Lee, Jong-Ho;Jo, Joung-Ae;Lee, Seung-Cheol;Lee, Suk-Keun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.5
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    • pp.440-453
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    • 2005
  • Objectives : To develop a bioactive membrane for guided bone regeneration (GBR), the biocompatibility and bone regenerating capacity of the cellulose membrane obtained from the Ascidians squirt skin were evaluated. Materials and methods : After processing the pure cellulose membrane from the squirt skin, the morphological study, amino acid analysis and the immunoreactivity of the cellulose membrane were tested. Total eighteen male Spraque-Dawley rats (12 weeks, weighing 250 to 300g) were divided into two control (n=8) and another two experimental groups (n=10). In the first experimental group (n=5), the cellulose membrane was applicated to the 8.0 mm sized calvarial bone defect and the same sized defect was left without cellulose membrane in the first control group (n=4). In the another experimental group (n=5), the cellulose membrane was applicated to the same sized calvarial bone defect after femoral bone graft and the same sized defect with bone graft was left without cellulose membrane in the another control group (n=4). Each group was sacrificed after 6 weeks, the histological study with H&E and Masson trichrome stain was done, and immunohistochemical stainings of angiogenin and VEGF were also carried out. Results : The squirt skin cellulose showed the bio-inductive effect on the bone and mesenchymal tissues in the periosteum of rat calvarial bone. This phenomenon was found only in the inner surface of the cellulose membrane after 6 weeks contrast to the outer surface. Bone defect covered with the bioactive cellulose membrane showed significantly greater bone formation compared with control groups. Mesenchymal cells beneath the inner surface of the bioactive cellulose membrane were positive to the angiogenin and VEGF antibodies. Conclusion : We suppose that there still remains extremely little amount of peptide fragment derived from the basement membrane matrix proteins of squirt skin, which is a kind of anchoring protein composed of glycocalyx. This composition could prevent the adverse immunological hypersensitivity and also induce bioactive properties of cellulose membrane. These properties induced the effective angiogenesis with rapid osteogenesis beneath the inner surface of cellulose membrane, and so the possibilities of clinical application in dental field as a GBR material will be able to be suggested.

Relationships between in-vitro virulence-associated characteristics, plasmid-bearing and production of Outer Membrane Protein(OMP) of Yersinia enterocolitica isolated from pigs (Yersinia enterocolitica의 시험관내 병원성 성상, plasmid 보유 및 외막 단백질(OMP) 생산간의 관계)

  • Park, Seog-gee;Choi, Chul-soon;Jeon, Yun-seong
    • Korean Journal of Veterinary Research
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    • v.32 no.2
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    • pp.181-194
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    • 1992
  • Two hundred and eighty nine strains of Yersinia enterocolitica isolated from healthy pigs were tested for the presence of 40~50 Megadalton virulence-associated plasmids and plasmidmediated in vitro virulence-associated properties, i.e., congo red uptake, calcium dependency, autoagglutination, CRMOX reaction, crystal violet binding and pyrazinamidase reaction. The correlationships between in vitro virulence-associated properties and the presence of 220 Kdalton outer membrane protein(OMP) were examined in strains with or without virulence-associated plasmids. The correlationships between the presence of plasmids on the production of the OMP and the expression of in vitro virulence-associated properties were studied with CRMOX+ strains and acridine orangecured CRMOX mutants. The results were as follows : 1. Of the in vitro virulence-associated tests with 289 strains of Y enterocolitica, 275 strains (95.2%) were positive for pyrazinamidase test, and followed by in order of crystal violet binding test, 226 (79.2% ) ; CRMOX test, 190 (65.7%) ; autoagglutination test, 1.85(64.0%) : calcium dependency test, 86 (29.8%) and congo red uptake test, 47(16.3%). 2. The correlationship between autoagglutination and CRMOX test(r=0.90) was highly significant (p<0.01). 3. In 190 strains(65.7%) bearing the virulence-associated plasmids(MW 40~50 Mdalton), the correlation between the presence of plasmids and their in vitro virulence-associated properties were highest with CRMOX test(r=0.93) and followed by in orders of AAG test(0.81), CV test(0.46), PYZ test(0.37) and CD test(0.18), but no correlationship between the presence of plasmids and CR test(-0.11). 4. The CRMOX+ strains produced the 220 Kdalton OMP when they were cultured at 37C, but not at 26C. The presence of 220 Kdalton OMP was correlated significantly with in vitro virulence properties and the presence of virulence-associated plasmid, respectively. 5. In the isogenic CRMOX mutant strains, of which plasmid were cured by treatment with acridine orange not only in vitro virulence-associated properties(CR 100%, CD 100%, AAG 82.6%, CV 58.3%) disappeared but also 220 Kdalton OMP(100%) was not produced. These results indicate that the positive CRMOX reaction is plasmid-mediated and the CRMOX test is potential as an in vitro virulence tests with Y enterocolitica.

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Efficient Delivery of Toxoid Antigens using Micro/Nano-carriers (마이크로/나노-운반체를 이용한 톡소이드 항원의 효과적인 전달 방법)

  • Park, Ga-Young;Ahn, Gna;Lee, Se Hee;Kim, Sang Bum;Kim, Yang-Hoon;Ahn, Ji-Young
    • Journal of Life Science
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    • v.28 no.4
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    • pp.496-507
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    • 2018
  • Immunization has been performed for centuries and is generally accepted as a sustainable method of controlling bacteria, viruses, and mediated and infectious diseases. Despite many studies having been performed on animal subjects to demonstrate the importance of toxin immunity, the use of toxoid vaccines in humans and animals has been limited for a long time. Recently, the development of the toxoid antigen delivery system has been facilitated using novel nano-medicinal technology. The micro/nano-carrier has been used to improve vaccination coverage as well as reduce vaccine costs. A micro/nano-carrier is a micro/nano-sized material that delivers immune cargo, including recombinant or peptide toxoid antigens. These toxoid antigens are either encapsulated in the interior or displayed on the surface of micro/nano-carriers as a way to protect them from the cellular machinery. In particular, the combination of toxoid antigens and micro/nano-carriers can induce phagocytosis through the specific interactions between GCs and macrophages; thus, the toxoid antigens can be delivered easily into the macrophages. This paper reviews recent achievements of micro/nano-carriers in the field of vaccine delivery systems such as microbial ghost cells (GCs, Bacterial ghost cells and Yeast ghost cells), gene-manipulated outer membrane vesicles (OMVs) and biocompatible, polymer-based nanoparticles (NPs, NP-Carrier and NP-Cage). Finally, this review shows various aspects in terms of the hosts' immune responses.