• Title/Summary/Keyword: intact membrane

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In Vivo Wound Healing Activity of Crocodile (Crocodylus siamensis) Hemoglobin and Evaluation of Antibacterial and Antioxidant Properties of Hemoglobin and Hemoglobin Hydrolysate

  • Pakdeesuwan, Anawat;Araki, Tomohiro;Daduang, Sakda;Payoungkiattikun, Wisarut;Jangpromma, Nisachon;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.1
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    • pp.26-35
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    • 2017
  • The hydrolysis of proteins constitutes an invaluable tool, granting access to a variety of peptide fragments with potentially interesting biological properties. Therefore, a hemoglobin (Hb) hydrolysate of Crocodylus siamensis was generated by digestion under acidic conditions. The antibacterial and antioxidant activities of the Hb hydrolysate were assessed in comparison with intact Hb. A disc diffusion assay revealed that the Hb hydrolysate exhibited antibacterial activity against eight strains of gram-positive bacteria and showed a higher efficacy than intact Hb. Moreover, the antioxidant activity of intact Hb and its hydrolysate was evaluated using ABTS and DPPH radical scavenging assays. The Hb hydrolysate exhibited free radical scavenging rates of 6-32%, whereas intact Hb showed a slightly higher activity. In addition, non-toxicity to human erythrocytes was observed after treatment with quantities of Hb hydrolysate up to $10{\mu}g$. Moreover, active fragmented Hb (P3) was obtained after purifying the Hb hydrolysate by reversed-phase HPLC. Scanning electron microscopy demonstrated the induction of bacterial cell membrane abnormalities after exposure to P3. Antibacterial and antioxidant activities play crucial roles for supporting the wound healing activity. Consequently, an in vivo mice excisional skin wound healing assay was carried out to investigate the effects of intact Hb treatment on wound healing in more detail. The results clearly demonstrate that intact Hb is capable of promoting 75% wound closure within 6 days. These findings imply that intact Hb of C. siamensis and its acid hydrolysate may serve as valuable precursors for food supplementary products benefitting human health.

Multiple Binding Affinities for Muscarinic Acetylcholine Receptors in Rat Brain (흰쥐 뇌내(腦內)의 무수카린성 콜린 수용체의 이질성(異質性))

  • Lee, Jong-Hwa;El-Fakahany, Esam E.
    • The Korean Journal of Pharmacology
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    • v.23 no.2
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    • pp.101-111
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    • 1987
  • We investigated the binding properties of $(^3H)$ QNB and $(^3H)$ NMS to mAchR to elucidate the characterstics of mAchR in rat brain by using two different preparations (homogemates & intact brain cell aggregates). The binding properties of both ligands demonstrated high affinity and saturability in both experiments, however $(^3H)$ QNB showed a significantly higher maximal binding capacity than tha ot $(^3H)$ NMS 1. In rat brain homogenates; Displacement of both lignands with several mAchR antagonists resulted in competition curves in accoradnce with the law of massaction for QNB, atropine & scopolamine in thie preparation, also a similar profile was found for the quaternary ammonium analogs of atropine & scopolamine (methyl atropine & methylscopolamine) when $(^3H)$ NMS was used to label the receptors in rat brain. But when these hydrophillic antagonists were used to displace $(^3H)$ QNB, they showed interaction with high- and low-affinity binding sites in brain homogenates. Pirenzepine, the nonclassical mAchR antagonist, was able to displace both ligands from binding sites in this preparation. 2. In intact rat brain cell aggregates; Intact bain cell aggregates were used to elucidate the binding characteristics of $(^3H)$ NMS to mAchR in rat. The magnitude of binding of this ligand was related linearly to the amount of cell protein in the binding assay with a high ratio of total to nonspecific binding. mAchR antagonists displaced specific $(^3H)$ NMS binding according to the law of mass-action, while it was possible to resolve displacement curves using mAchR agonist into high-& low-affinity component. 3. Our results indicate that more hydrophilic receptor ligand $(^3H)$ QNB, displacement experiments in both tissues demonstrated that the lipid solubility of a particulr mAchR ligand might play an important role in determining its profile of binding to the mAchR, and the concentrations of mAchR in rat brain are both on the cell surface (membrane-bound receptor) and in the intracelluar membrane (intermembrane-bound receptor). 4. The results are discussed in terms of the usefulness of dissociated intact rat brain cells in studying mAchR in central nervous system.

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Effects of Caponization on Bone Characteristics and Histological Structure in Chickens

  • Chen, Kuo-Lung;Chang, Ming-Huang;Tsay, Shiow-Min;Hurng, Huaang-Youh;Chiou, Peter Wen-Shyg
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.2
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    • pp.245-251
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    • 2006
  • The aim of this study was to investigate the effects of caponization on the bone characteristics, biomechanical property and histology in Taiwan country chickens fed to market age of 26 wks. Male Taiwan country chickens $D{\times}L_2$ were caponized or sham-operated at 8 wks of age, and selected healthy sham-operated and completely caponized chickens (prominent degenerated comb) were selected at 16 wks old and fed to 26 wks old for the trials. Fifteen intact male chickens (Intact), sham-operated chickens (Sham) and caponized chickens (Capon) were assigned for trial 1, and sixteen Intact and Capon were assigned for trial 2. Results in trial 1 showed that the abdominal fat and relative abdominal fat weights of Capon were significantly heavier than Intact and Sham (p<0.05), while the tibia weight and relative weight were the lightest (p<0.05). The tibia breaking strength, bending moment and stress of Capon were the poorest among groups (p<0.05). The trial 2 produced the similar observation that Capon were significantly lighter than Intact (p<0.05) in the tibia weight, relative tibia weight and their biomechanical properties. On histological determinations, Capon showed a thinner cartilage end and fewer chondrocytes (about 50%) and trabecular, and bigger marrow cavity; while decreased hemopoietic cells number with increased adipocytes than Intact observed by H&E stain and at low magnification. At high magnification, Capon showed a decrease in the chondrocyte size by 33 to 50%, with smaller nucleus located near the cell membrane, and exhibited monocellular form chondrocytes. Capon also showed a less strongly acidic sulfated mucosubstance with weaker dyeing property within cartilage zone, and smaller chondrocytes size by Alcian blue stain.

Study of the ageing of hollow fibers in an industrial module for drinking water production

  • Wang, S.;Wyart, Y.;Perot, J.;Nauleau, F.;Moulin, P.
    • Membrane and Water Treatment
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    • v.4 no.1
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    • pp.53-67
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    • 2013
  • In this study, ageing characteristics of an industrial hollow-fiber membrane module were investigated after 50 months of drinking water production. For this purpose, the industrial module was opened to make 18 smaller modules with hollow-fibers taken from different parts of the industrial module. These modules were probed by the use of a magnetic nanoparticle (NP) challenge test based on magnetic susceptibility (K) measurement of permeate. No magnetic susceptibility was detected in permeate when the challenge test was performed on an intact membrane module, indicating the complete retention of nanoparticles by the membrane. The compromised membrane module can be successfully detected by means of magnetic susceptibility measurement in permeate. So, this study clearly demonstrates that ageing of ultrafiltration membranes can be monitored by measuring the magnetic susceptibility of permeate from an ultrafiltration membrane module. These results showed that the hollow fibers in the center zones of the bundle would age faster than those in the outer zones around the bundle. This result is in agreement with numerical simulation (Daurelle et al. 2011).

Expression of Latent P-Type ATPases and Their Presumptive Roles in Cell Membrane of Helicobacter pylori

  • YUN, SOON-KYU;SE-YOUNG HWANG
    • Journal of Microbiology and Biotechnology
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    • v.7 no.6
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    • pp.378-385
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    • 1997
  • Cation motive ATPases on cell membranes of Helicobacter pylori were investigated using everted membrane vesicles. Latent ATPases could be ascertained from aggregated vesicle using N, N-dimethylformamide (DMF) and Triton X-100. By contrast, ultrasonication or chloroform treatments caused membranes to be disrupted, resulting in an alteration of sensitivities against azide or vanadate. Considerable amounts of vanadate-sensitive enzymes were identified from vesicle micelles, prepared by the dilution method. These were activated in the presence of either $Ni^{2+}\;or\;NH_4^+$. From studies employing H. pylori intact cell systems, we found that ATPase expression of this bacterium was markedly dependent upon air composition. It was interesting that cellular expression of $Ni^{2+}$- or $NH_4^{+}$-motive ATPases was significantly affected by extracellular pH, suggesting that these unique enzymes may physiologically be involved in cellular $Ni^2$ import and $NH_4^+$ export, respectively.

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Transdermal Delivery of Ethinylestradiol UsingEthylene-vinyl Acetate Membrane

  • Shin, Sang-Chul;Byun, Soo-Young
    • Journal of Pharmaceutical Investigation
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    • v.25 no.3
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    • pp.53-59
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    • 1995
  • Ethinylestradiol (EE)-containing matrix was fabricated with ethylene-vinyl acetate(EVA) copolymer to control the release of the drug, Effect of addition of PEG 400 as receptor solution, the stripping of skin and Azone pretreatment on skin on the permeation of EE through the excised mouse skin was also studied. The permeation rate of EE through the excised mouse skin was affected by the PEG 400 volume fraction. The Azone pretreatment on skin didn't affect on the steady state flux, however, the lag time was shortened. The permeation rate of EE through the stripped skin was much larger than that through the whole skin. It showed that the stratum corneum acts as a barrier of skin permeation. The fact that there is little difference in EE permeation between the intact skin and the stripped skin with EVA membrane shows the permeation of EE through the mouse skin is mainly controlled by the membrane.

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Effects of Light on the Pigment Production and Chloroplast Development of Ginseng Hairy Roots (인삼 모상근의 색소 생성 및 엽록체 발달에 미치는 광의 효과)

  • 양덕조;최혜연
    • Journal of Ginseng Research
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    • v.21 no.1
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    • pp.28-34
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    • 1997
  • The effects of light on the pigment production and chloroplast development were examined on ginseng hairy roots cultured in 1/2MS liquid medium. The chlorophyll and carotenoid production were increased from 1,000 to 3,500 lux condition, but decreased drastically in 7,000 lux condition. The anthocyanin production was significantly increased with increment light intensity(1,000∼7,000 lux). The thylakoid membrane of chloroplast was proplastid in dark condition and it began to develop into thylakoid membrane in 1,000 lux condition and then intact thylakoid membrane was developed in 3,500 lux condition. However, the development of thylakoid membrane in 7,000 lux condition was inhibited comparing to 3,500 lux condition. The total chlorophyll production in blue light condition were high comparing to other wavelength and same as 40% of total chlorophyll on white light(3,500 lux) condition. The chlorophyll and carotenoid production by sucrose concentration were high in 3% sucrose condition and anthocyanin production was high in 4% condition. The production of chlorophyll and carotenoid by light periods was high when explants were cultured in dark condition for 1 week and then transferred to light condition for 4 weeks. Our results suggest that pigment production and chloroplast development could be accelerated by light Intensity of specific wavelength in cultures of ginseng hairy root.

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Enterobacter cloacae MG82에 의한Triphenylmethane흡수 특성과 탈색효소의 세포내 위치

  • Jeong, Min-Seon;Kwak, Soon-Jun;Kim, Byung-Hong;Chung, Young-Gun;Kang, Sa-Ouk;Min, Kyung-Hee
    • Microbiology and Biotechnology Letters
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    • v.25 no.1
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    • pp.37-43
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    • 1997
  • Triphenylmethane was decolorized rapidly by enterbacter cloacae MG 82 at initial reaction time. The spheroplast showed higher activity of triphenylmentane decolorization than that of intact cell suspension. The outer part of the bacterial cell envelope and the peptidoglycan are important for the function of transport barrier of triphenylmethane. In intact cell, decolorization activity was higher at 37$\circ $C than at $\circ $C, indicating that triphenylmethane decolorization is due to the enzyme reaction. Culture filtrate showed no decolorization activity, while cell-free extract appeared high activity of 1.45 units, clearly showing that decolorization activity was due to the cell-free extract. Comparing decolorization activities of cell fractions, it was found that decolorization activity was located at the compartment of cytoplasmic membrane. The enzyme activity was also shown to be Mg$^{++}$-dependent. The optimum pH and temperature of enzyme activity were 7.0 and 50$\circ $C, respectively. The thermostability of this enzyme at 35$\circ $C was kept to 58% for 3 hours.

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Inhibitory Action of the Paraquat on Superoxide Dismutase of Excherichia coli (Paraquat에 의한 Escherichia coli의 Superoxide Dismutase 활성저해)

  • 김미림;최경호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.5
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    • pp.849-855
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    • 1994
  • Actively growin Excherichia coli(KCTC 1039) cells were treated with paraquat (1, 1'-dimethyl-4, 4'-bipyridili-um dichloride) by cultivating them in the presence of 1.0mM paraquat. The treatment was carried out with or without shaking to understand the effect of oxygen on paraquat action to thebacterial superoxide dismutase (SOd). By the treatment with vigorous shaking , population growth of the organism almostly stopped and specific activities of SOD of the cells drastically decreased. On contrast ot it, the herbicide showed only l limited inhibitory action on bacterial growth and SOD activity by stationary treatment. Proteins prepared from parquat-treated cells divided into two peaks by Sephacryl column chormatogrpahy, while proteins from the intact cells formed a single peak. Cytoplasmic proteins and plasma membrane proteins of intact cells formed separated three peaks by Sephadex G-75 column chormatography. respectively. Among them the second peak disappeared by paraquat treatment , while the third peak became more apparent. Fractions from the first and the third peak showed SOD activity. Paraquat was detected from the same fractions.

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Effects of Barbiturates on the Rotational Relaxation Time of 1, 6-Diphenyl-1, 3, 5-hexatriene in Native and Model Membranes

  • Chung, Yong-Za;Shin, Yong-Hee;Choi, Chang-Hwa;Park, Hyung-Sook;Koh, Yeong-Sim;Yun, Il
    • Archives of Pharmacal Research
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    • v.15 no.4
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    • pp.298-303
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    • 1992
  • Synaptosomal plasma membrane vesicles (SPMV) were isolated from fresh bovine cerebral cortex. The effects of barbiturates on the rotational relaxation time of 1.6-diphenyl-1, 3, 5-hexatriene (DPH) in intact SPMV and model membranes of total lipids (SPMVTL) and phosphlipids (SPMVPL) extracted from SPMV were examined. Barbiturates decreased the rotational relaxation time of DPH in intact SPMV in a dose-dependent manner. In contrast, they did not affect the rotational relaxation time of DPH in SPMVTL and even dose-dependently increased the rotational relaxation time of DPH in SPMVPL.

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