• Title/Summary/Keyword: denaturation

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Enzymatic Hydrolysis of Gelatin Layers of X-Ray Films and Release of Silver Particles Using Keratinolytic Serine Proteases from Purpureocillium lilacinum LPS # 876

  • Cavello, Ivana A.;Hours, Roque A.;Cavalitto, Sebastian F.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.8
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    • pp.1133-1139
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    • 2013
  • Enzymatic decomposition of gelatin layers on used X-ray films and repeated utilization of the enzyme for potential application in silver recovery were investigated using keratinolytic serine proteases from Purpureocillium lilacinum LPS # 876. At pH 9.0, the enzymatic reaction was enhanced by the increase of enzyme concentration or by the increase of the temperature up to $60^{\circ}C$. Under the conditions of 6.9 U/ml, $60^{\circ}C$, and pH 9.0, hydrolysis of the gelatin layers and the resulting release of silver particles were achieved within 6 min. The protective effect of polyols against thermal denaturation was investigated. The presence of glycerol and propylene glycol increased enzyme stability. When the reusability of the enzyme for gelatin hydrolysis was tested, it could be seen that it could be effectively reused for more cycles when glycerol was added, compared with the enzyme without protective agents. The results of these repeated treatments suggested that a continuous process of recycling silver from used X-ray is feasible. Keeping in mind that recycling is (at the present time) needed and imperative, it can be remarked that, in this research, three wastes were successfully used: hair waste in order to produce serine proteases; glycerol in order to enhance enzyme thermal stability; and used X-ray films in order to recover silver and PET films.

Sustained Release Injectable of Recombinant Bovine Somatotropin in Biodegradable Poly(D,L-lactide-co-glyceride) Microspheres (생분해성 폴리락티드/글리콜리드 미립구를 이용한 재조합 소 성장호르몬(rBST)의 지속성주사제 설계)

  • Jeon, Hong-Ryeol;Lee, Bong-Sang;Kown, Do-W;Yoon, Mi-Kyoung;Jeon, Hyun-Joo;Shin, Taek-Hwan;Choi, Young-Wook
    • Journal of Pharmaceutical Investigation
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    • v.32 no.3
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    • pp.199-207
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    • 2002
  • In order to develop a sustained release formulation of bovine somatotropin (BST), which has been used to increase the body weight of oxen or the milk production of dairy cows, poly(D,L-lactide-co-glyceride)(PLGA) microspheres were made by W/O/W multiple emulsification method and solvent extraction method. Physical properties including particle size, drug entrapment, drug release, protein denaturation, and in vivo body weight increase in rats were characterized. The size of the microspheres was increased as the molecular weight of PLGA increased. When Span 65 and stearic acid during preparation were added, the size was decreased but the amount of surface protein was increased, resulting in a high loading efficiency, with fast release of BST from the microspheres. Aggregation or fragmentation of BST by SDS-PAGE during microsphere preparation and drug release study was not observed. Body weight of Sprague-Dawley's male rats was significantly increased after subcutaneous administrations of BST-loaded PLGA microspheres. There was a good correlation between in vivo weight gain and in vitro release rate of microspheres. PLGA microspheres with a high surface protein ratio could be a good candidate for the sustained delivery of BST.

Thermally Induced Gelation of Alaska Pollack Meat Paste -1. Effects of NaCl and Starch on the Thermal Properties of Alaska Pollack Meat Paste- (가열에 의한 명태 연육의 Gel화에 관한 연구 -1. 명태 연육의 열특성에 미치는 식염과 전분의 영향-)

  • JUNG Woo-Jin;PARK Seong-Min;LEE Kang-Ho;LEE Keun-Tai
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.1
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    • pp.21-25
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    • 1993
  • In order to obtain the fundamental factors influencing on gelation of Alaska pollack meat paste during processing, thermograms of protein using differential scanning calorimetry (DSC) were investigated. The thermal transition temperatures of Alaska pollack meat paste due to protein denaturation were $38^{\circ}C,\;49^{\circ}C,\;55^{\circ}C\;and\;77^{\circ}C$, but those temperatures were changed to $35^{\circ}C,\;45^{\circ}C,\;50^{\circ}C\;and\;73^{\circ}C$ after adding salt($3\%$ NaCl). The starch did not affect the thermal transition of fish protein and its thermal properties were changed independently in starch-meat paste mixture system.

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Effects of Kanagawa Hemolysin on Blood Pressure and Arterial Tone in Rats

  • Kim, Young-Moon
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.4
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    • pp.225-233
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    • 2002
  • Kanagawa hemolysin (KH), an exotoxin produced from Kanagawa phenomenon-positive Vibrio parahemolyticus, has been shown to possess various biological activities including hemolysis, enterotoxicity, cytotoxicity, and cardiotoxicity. The aim of this study was to investigate the effect of KH on the cardiovascular system and its mechanism, employing in vivo and in vitro experiments of the rat. Intracerebroventricular (icv) administration of 100 mHU KH produced a marked and continuous pressor effect (icv KH-pressor effect), and the icv pressor effect was not repeatable. However, intravenous (iv) injection of the same dose of KH induced a prominent depressor effect (iv KH-depressor effect). The icv KH-pressor effect was inhibited by acid-denaturation, while the iv KH-depressor effect was not. Simultaneous icv administration of the three agents (ouabain, diltiazem, or bumetanide: $10{\mu}g/kg$ each) significantly reduced the pressor effect. The icv KH-pressor effect was inhibited by treatment with iv phentolamine or chlorisondamine, but was not affected by iv candesartan. The iv KH-depressor effect was repeatable and was attenuated by treatment with iv NAME or methylene blue. In vitro experiments using isolated thoracic aorta, $10^{-6}$ M phenylephrine (PE) and 50 mM KCl produced a sustained contraction. In rings contracted with either agents, KH showed relaxant responses in a concentration- dependent fashion and the relaxation (KH-vasorelaxation) was not dependent on the existence of the endothelium. The KH-vasorelaxation in the endothelium-intact rings contracted by PE was abolished by methylene blue treatment. In summary, the present findings suggest that in the icv KH-pressor effect the cation leak-inducing action of KH is implicated, which leads to the increased central sympathetic tone, that the iv KH-depressor effect results from the vasorelaxation via NO-guanylate cyclase system, and that the KH-vasorelaxation is independent of the endothelium and the guanylate cyclase system is involved in it. In conclusion, the mechanism of KH producing the icv pressor effect may not be identical to that of KH producing the iv depressor effect.

Roles of YehZ, a Putative Osmoprotectant Transporter, in Tempering Growth of Salmonella enterica serovar Typhimurium

  • Kim, Seul I;Ryu, Sangryeol;Yoon, Hyunjin
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1560-1568
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    • 2013
  • Salmonella, a main cause of foodborne diseases, encounters a variety of environmental stresses and overcomes the stresses by multiple resistance strategies. One of the general responses to hyperosmotic stress is to import or produce compatible solutes so that cells maintain fluid balance and protect proteins and lipids from denaturation. The ProP and ProU systems are the main transport systems for compatible solutes. The OsmU system, recently identified as a third osmoprotectant transport system, debilitates excessive growth as well by reducing production of trehalose. We studied a fourth putative osmoprotectant transport system, YehZYXW, with high sequence similarity with the OsmU system. A Salmonella strain lacking YehZ, a predicted substrate-binding protein, did not suffer from hyperosmolarity but rather grew more rapidly than the wild type regardless of glycine betaine, an osmoprotectant, suggesting that the YehZYXW system controls bacterial growth irrespective of transporting glycine betaine. However, the growth advantage of ${\Delta}yehZ$ was not attributable to an increase in OtsBA-mediated trehalose production, which is responsible for the outcompetition of the ${\Delta}osmU$ strain. Overexpressed YehZ in trans was capable of deaccelerating bacterial growth vice versa, supporting a role of YehZ in dampening growth. The expression of yehZ was increased in response to nutrient starvation, acidic pH, and the presence of glycine betaine under hyperosmotic stress. Identifying substrates for YehZ will help decipher the role of the YehZYXW system in regulating bacterial growth in response to environmental cues.

Conformational Study of Human Serum Albumin in Pre-denaturation Temperatures by Differential Scanning Calorimetry, Circular Dichroism and UV Spectroscopy

  • Rezaei-Tavirani, Mostafa;Moghaddamnia, Seyed Hassan;Ranjbar, Bijan;Amani, Mojtaba;Marashi, Sayed-Amir
    • BMB Reports
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    • v.39 no.5
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    • pp.530-536
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    • 2006
  • Thermal conformational changes of human serum albumin (HSA) in phosphate buffer, 10 mM at pH = 7 are investigated using differential scanning calorimetric (DSC), circular dichroism (CD) and UV spectroscopic methods. The results indicate that temperature increment from $25^{\circ}C$ to $55^{\circ}C$ induces reversible conformational changes in the structure of HSA. Conformational change of HSA are shown to be a three-step process. Interestingly, melting temperature of the last domain is equal to the maximum value of fever in pathological conditions, i.e. $42^{\circ}C$. These conformational alterations are accompanied by a mild alteration of secondary structures. Study of HSA-SDS (sodium dodecyl sulphate) interaction at $45^{\circ}C$ and $35^{\circ}C$ reveals that SDS affects the HSA structure at least in three steps: the first two steps result in more stabilization and compactness of HSA structure, while the last one induces the unfolding of HSA. Since HSA has a more affinity for SDS at $45^{\circ}C$ compared to $35^{\circ}C$, It is suggested that the net negative charge of HSA is decreased in fever, which results in the decrease of HSA-associated cations and plasma osmolarity, and consequently, heat removal via the increase in urine volume.

Recyclable Polymeric Composite with High Thermal Conductivity (재활용 가능한 고방열 고분자 복합소재 개발)

  • Shin, Haeun;Kim, Chae Bin;Ahn, Seokhoon;Kim, Doohun;Lim, Jong Kuk;Goh, Munju
    • Composites Research
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    • v.32 no.6
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    • pp.319-326
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    • 2019
  • To address tremendous needs for developing efficiently heat dissipating material with lightweights, a new class of polymer possessing recyclable and malleable characteristics was synthesized for incorporating model functional hexagonal boron nitride (h-BN) filler. A good interfacial affinity between the polymer matrix and the filler along with shear force generated upon manufacturing the composite yielded the final product bearing highly aligned filler via simple hot pressing method. For this reason, the composite exhibited a high thermal conductivity of 13.8 W/mK. Moreover, it was possible to recover the h-BN from the composite without physical/chemical denaturation of the filler by chemically depolymerizing the matrix, thus the recovered filler can be re-used in the future. We believe this polymer could be beneficial as matrix for incorporating many other functional fillers, thus they may find applications in various polymeric composite related fields.

Development of Human-Head-Mimicking Phantom for Brain Treatment Using Focused Ultrasound (집속 초음파 뇌 질환 치료를 위한 두부 유사 팬텀의 개발)

  • Min, Jeonghwa;Kim, Juyoung;Noh, Sicheol;Choi, Heungho
    • Journal of the Korean Society of Radiology
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    • v.7 no.6
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    • pp.433-439
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    • 2013
  • In this study, human head-mimicking phantom was developed for brain disease treatment study using focused ultrasound. Acoustic parameters of skin, skull and brain were investigated through literature investigation and adequate substitutes according to each tissue were suggested. In the case of skin phantom, construction ratio of glycerol-based TMM phantom was controlled to mimic real skin. The suitability of skull substitutes was evaluated through measurement of acoustic parameters. In the case of brain phantom, transparent egg white phantom was used to observe thermal properties of focused ultrasound. Combined human-head-mimicking phantom using each substitutes was fabricated for development of brain disease treatment protocol. Denaturation of brain phantom according to ultrasonic condition was observed for validation.

Viscosity of Yam Suspension by Drying Methods and Additives (건조방법과 첨가물에 따른 마 현탁액의 점도)

  • Kim, In-Hyun;Son, Hyun-Ju;Chung, Koo-Min
    • Korean Journal of Food Science and Technology
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    • v.38 no.3
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    • pp.444-447
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    • 2006
  • Yam powders were prepared by freeze, fan, hot-air, and coal-heat drying. The viscosities of their suspensions and supernatants and the viscosity changes with addition of sugar, salt, and citric acid were investigated. Viscosity (43 mPa s) of 7.5% suspension of fan-dried yam powder was lower than that of freeze dried yam (58.1 mPa s), but much higher than that of the conventional, hot-air dried yam (17.2 mP s). Coal-heat dried yam had a viscosity of only 4.5 mPa s. The viscosity was related to the protein denaturation induced by heat and acid. Addition of sugar to yam powder increased the viscosity of the suspension but no changes were evident with salt addition. Viscosities decreased when 0.5% citric acid was added (pH 3.4-3.5).

Cloning of Bovine Macrophage Colony-stimulating Factor

  • Kim, Tae-Yung;Kim, Cheol-Ho;Lee, Sang-Gil;Kang, Chung-Boo
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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    • pp.892-897
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    • 2005
  • Macrophage colony-stimulating factor (M-CSF) is a growth factor required for growth and differentiation of mononuclear phagocyte lineage. Total and 16 poly (A) mRNA of bovine M-CSF were isolated from healthy bovine peripheral mononuclear cells stimulated by phobol 12-myristste 13-acetate (TPA). The more compatible cultured mononuclear cells were 5${\times}$10/ml for RNA isolation. TPA-activated mononuclear cells increased the level of M-CSF-mRNA more than concanavalin A (Con A) and lipopolysaccharide (LPS). The optimal analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) for14 Macrophage colonystimulating factor (M-CSF) as a growth factor required for bovine M-CSF was denaturation at 94$^{\circ}C$ for 1 minute, annealing at 57$^{\circ}C$ for 1 minute, extension at 72$^{\circ}C$ for 1 minute for 30 cycles. The size of cDNA of bovine M-CSF by RT-PCR was 774 base pairs. A 774 base pairs cDNA encoding bovine M-CSF was synthesized by reverse transcriptase polymerase chain reaction (RT-PCR). Ligated cDNA was transformed to competent cells and then plasmid isolation and digestion was performed. Molecular cloning and sequencing were performed for cDNA of bovine M-CSF. The size of cloned cDNA of bovine M-CSF was 774base pairs. The homology of base sequence and amino acid sequence was 88% and 86% compared with known human M-CSF, respectively. From a high degree of sequence similarity, the obtained cDNA of bovine M-CSF is thought be a specific gene of bovine M-CSF.