• Title/Summary/Keyword: protein heat treatment

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Effects of a mild heat treatment on mouse testicular gene expression and sperm quality

  • Zhao, Jun;Zhang, Ying;Hao, Linlin;Wang, Jia;Zhang, Jiabao;Liu, Songcai;Ren, Bingzhong
    • Animal cells and systems
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    • v.14 no.4
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    • pp.267-274
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    • 2010
  • The decrease in sperm quality under heat stress causes a great loss in animal husbandry production. In order to reveal the mechanism underlying the sperm quality decrease caused by heat stress, we first established a mild heat-treated mouse model. Then, the sperm quality was identified. Further, the testicular proteome profile was mapped and compared with the control using 2D electrophoresis and mass spectrometry. Finally, the differential expressed proteins involved in the heat stress response were identified by real-time PCR and Western blotting. The results showed that heat stress caused a significant reduction in mouse sperm quality (P<0.05). Further, 52 protein spots on the 2D gel were found to differ between the heat-shocked tissues and the control. Of these spots, some repair proteins which might provide some explanation for the influence on sperm quality were found. We then focused on Bag-1, Hsp40, Hsp60 and Hsp70, which were found to be differently expressed after heat shock (P<0.05). Further analysis in this heat-shocked model suggests numerous potential mechanisms for heat shock-induced spermatogenic disorders.

Rescue of Oxidative Stress by Molecular Chaperones in Yeast

  • Ueom Jeonghoon;Kang Sooim;Lee Kyunghee
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.76-78
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    • 2002
  • Heat shock proteins (HSPs) are induced in most living cells by mild heat treatment, ethanol, heavy metal ions and hypoxia. In yeast Saccharomyces cerevisiae, mild heat pretreatment strongly induces Hsp104 and thus provide acquired thermotolerance. The ability of hsp104 deleted mutant $({\triangle}hsp104)$ to acquire tolerance to extreme temperature is severely impaired. In providing thermotolerance, two ATP binding domains are indispensible, as demonstrated in ClpA and ClpB proteases of E. coli. The mechanisms by which Hsp104 protects cells from severe heat stress are not yet completely elucidated. We have investigated regulation of mitochondrial metabolic pathways controlled by the functional Hsp104 protein using $^{13}C_NMR$ spectroscopy and observed that the turnover rate of TCA cycle was enhanced in the absence of Hsp104. Production of ROS, which are toxic to kill cells radiply via oxidative stress, was also examined by fluorescence assay. Mitochondrial dysfunction was manifested in increased ROS levels and higher sensitivity for oxidative stress in the absence of Hsp104 protein expressed. Finally, we have identified mitochondrial complex I and Ferritin as binding protein(s) of Hsp104 by yeast two hybrid experiment. Based on these observations, we suggest that Hsp104 protein functions as a protector of oxidative stress via either keeping mitochondrial integrity, direct binding to mitochonrial components or regulating metal-catalyzed redox chemistry.

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Reduced Protein Denaturation in Thermotolerant Cells by Elevated Levels of HSP70 (열내성이 유도된 세포에서 HSP70 단백질 증가에 의한 단백질 변성 감소)

  • Han, Mi-Young;Park, Young-Mee
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.433-444
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    • 1996
  • We describe a novel approach to evaluate quantitatively the amounts of denatured proteins in cells upon heat exposure. A thiol compound, diamide [azodicarboxylic acid bis (dimethylamide)] causes protein cross-linking with exposed sulfyhydryl residues of denatured proteins. Since denatured proteins expose normally well-hidden sulfhydryl groups, these will be preferentially cross-linked by diamide. Thus diamide acts to 'trap' denatured proteins. We observed that protein aggregates (high molecular weight protein aggregates, HMA) appeared on SDS-polyacrylamide gels run under non-reducing conditions and that the amount of HMA can be quantified by scanning the gels using a gas flow counter. Heating cells followed by a fixed dose of diamide exposure resulted in HMA increases in a heat-dose dependent manner, demonstrating that the quantitation of HMA could serve as a measure of heat-denatured proteins. We compared thermotolerant and nontolerant cells and found decreased HMA in tolerant cells upon heat treatment. As an attempt to examine the kinetics of protein renaturation (or 'repair'), we measured the amounts of aggregates formed by the addition of diamide at various times after heat shock. Such experiments demonstrate an equally rapid disappearance of HMA in previously unheated and in thermotolerant cells. Levels of HMA in tolerant cells increased significantly after electroporation of HSP70 specific mAbs, suggesting an involvement of HSP70 in reducing HMA levels in thermotolerant cells upon heat exposure. Immunoprecipitation studies using anti-HSP70 antibody indicated an association of HSP70 with heat-denatured proteins. Our results suggest that heat induces protein denaturation, and that elevated level of HSP70 present in thermotolerant cells protects them by reducing the level of protein denaturation rather than by facilitating the 'repair' (or degradation) process.

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A study on the changes of the components in the steaming and roasting green tea after heat treatments according to time. (가열처리시간이 Steaming 및 Roasting green tea의 성분변화에 미치는 영향)

  • 고영수
    • Journal of the Korean Home Economics Association
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    • v.23 no.2
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    • pp.29-36
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    • 1985
  • This experiment was conducted to investigate changes of the components in steamed and roasted green tea after 10 min, 20 min and 30 min of heat treatment of heat treatment at $110^{\circ}C$. Vitamin C and tannin were determined by spetrophotometry. Total nitrogen, water-soluble nitrogen and water-soluble non protein nitrogen were determined by Kjeldahl analysis system. Aromas were identified by gas chromatography. After heat treatment, contents of most compounds decreased. Tannin and caffeine decreased a little bit, while vitamin C decreased considerably. There were both decreases and increases in aromas. Aromas which increased were geraniol, trans-2-hexenol, linalool, $\alpha$-ionone and $\beta$-ionone. From these results, it was concluded that the highest quality of green tea was one steam- treated for 10 min at $110^{\circ}C$.

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Effects of Protease Treatment and Animal Behavior on the Dissociative Culture of Aplysia Neurons

  • Lee, Nuribalhae;Rim, Young-Soo;Kaang, Bong-Kiun
    • Animal cells and systems
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    • v.13 no.3
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    • pp.267-274
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    • 2009
  • The dissociative culture technique of Aplysia neuron is one of the key methods that have been used for studies of cellular and molecular mechanisms of neuronal functioning. However, despite the advantages this method offers as an experimental model, its technical efficiency has had room for improvement. In this study, we examined certain putative factors that might affect the culture quality. The effects of neuronal damage induced by physical injuries, heat shock, and surface protein degradation were evaluated along with the correlation between the culture quality and animal behavior. As a result, we found that physical injury can be a critical factor that affects culture quality, whereas the heat shock and surface protein degradation had negligible effect on it. In addition, we discovered that siphon retraction time was not a good measurement for healthy neurons. Based on these findings, we suggest here an improved method in which the degree of physical injury is reduced by means of multiple protease treatment.

Effects of Cadmium on Heat Shock Protein Induction and on Clinical Indices in Rats (카드뮴이 랫드의 Heat Shock Protein 발현에 미치는 영향과 독성학적 변화에 관한 연구)

  • 김판기
    • Journal of Environmental Health Sciences
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    • v.22 no.4
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    • pp.91-101
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    • 1996
  • Exposure indices are important tools which enable scientists to reliably predict and detect exposures to xenobiotics and resultant cell injury. Since the de novo synthesis of stress proteins can be detected early after exposure to some agents, analysis of toxicant-induced changes in gene expression, i.e. alterations in patterns of protein synthesis, may be useful to develop as biomarkers of exposure and toxicity. The acute and chronic effects of cadmium(Cd, $CdCl_2$ 20 mg/kg) on Wistar male rats were evaluated concerning cadmium contents, tissues enzyme activity, HSP expression. The results of the study were as follows: 1. Less cadmium was absorbed through the digestive tracts, but the ratio of contents in renal to hepatic cadmium was higher at 8 weeks after treatment. 2. ALT(alanine aminotransferase), AST(aspartate aminotransferase), glucose, BUN(blood urea nitrogen), creatinine, the key indices of the clinical changes in hepatic and renal function were significantly changed by the cadmium treatment after 1 week in liver, after 4 weeks in kidney. 3. Enhanced synthesis of 70 KDa relative molecular mass proteins were detected in 2 hours after cadmium exposure, with maximum activity occurring at 8~48 hours. Induction of $HSP_{70}$ was evident at proximal tubules and glomeruli in kidney. Testicular cells produced enough HSP to be detected normally. From the above results, it could be concluded that $HSP_{70}$ induction by the cadmium treatment was a rapid reaction to indicate the exposure of xenobiotics.

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흰쥐 적출대동맥의 수축력에 미치는 열과 Nacl의 영향

  • Park Tae Gyu;Kim Jong Il;Seong Yu Jin;Kim In Gyeom;Kim Jung Yeong
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2003.11a
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    • pp.86-91
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    • 2003
  • In this study, in order to examine whether salt and heat shock stress would alter or not contraction and relaxation of isolated rat aorta. Under anesthesia with sodium pentobarbital(50 mg Kg$^{-1}$ i.p.), male Sprague Dawley rats weighing 300-330 g were subjected to 0, heat shock combined salt stress, where as the sham group was left at modified Krebs-bicarbonate solution. To measure contractile response of vascular ring preparation isolated from rat was determined in organ bath and was recorded on physiograph connected to isometric transducer. And the strip was checked for expression of heat shock protein(Hsps) by means of western blotting. The combination group of heat and 50 mM NaCl group increased vascular contractility, and the heat and 150 mM NaCl group decreased vascular contractility for 5 hours, and then recovered for 8 hours compared to that of control. Expressin of Hsp 70 of vascular muscle of rat aorta more increased by combination of heat and NaCl treatment than those of single treatment of heat or NaCl treatment, and vascular Hsp 70 showed a little decrease at 8 hours compared at 5 hours. These result indicate that mixed environmental stress either increased or decreased in vascular contractility by combination of heat and NaCl concentration.

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Development of Leaf Protein Concentrates I. Studies on the Isolation of Leaf Protein Concentrates (잎 단백질(蛋白質)(Leaf Protein Concentrates)의 개발(開發)에 관한 연구(硏究) -I. 잎 단백질(蛋白質)의 추출조건(抽出條件)에 대한 검토(檢討)-)

  • Choe, Sang;Kim, Ceon-Chee;Chun, Myung-Hi;Kim, Kil-Hwan
    • Korean Journal of Food Science and Technology
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    • v.2 no.2
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    • pp.8-16
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    • 1970
  • Exploitation of leaf protein concentrates for human consumption is very important. Leaf protein concentrates can be easily prepared by mechanically mincing leaves material and press it for getting the juice. Crude protein can be separated from the juice by aging, adjusting the pH, or heating to $75-80^{\circ}C$ etc. This report deals with the extractability of total-N from 69 species of fresh leaves by mechanical process, and then compared the recovery of leaf protein concentrates from leaf extracts by treating with TCA, pH adjustment and heating. Results are summarized as follows. 1. In general, the greater the content of total-N of leaves the greater the percentage extraction. Extraction of the juice from leaves is needed at least two times. The simple equations are constituted between the total-N (T; %) and the first and second extractability ($E_1,\;E_2;\;%$) of the total-N of leaves, as follows: $E_1=0.8168T\;E_2=0.1830T$ 2. The optimum pH value for coagulating protein from extracts is considered to be 3.5 to 4.5. However, the products of leaf protein concentrate by the pH adjustment of extracts are generally dull in color with rich elasticity. 3. Recoveries of the leaf protein concentrate from extracts by treating methods were in the following order of TCA treatment> pH 4 treatment> pH 3 treatment> heat treatment. The yield of leaf protein concentrates decreased bout 10% with pH 4 treatment, 11.4% with pH 3 treatment, and 14.8% with heat treatment compared with the TCA treatment. 4. The heat treatment is the most benifitial method for the production of leaf protein concentrates with regard to properties of texture, color and yield of products and easiness of the treatment method.

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Scientific exploration on physiological basis of Svedana Karma (Sudation): A clinical application of heat stress.

  • Yadav, Saurabh;Verma, Vandana;Abhinav, Abhinav
    • CELLMED
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    • v.9 no.3
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    • pp.4.1-4.8
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    • 2019
  • Now researchers have focused attention on exploring the mechanism of acute responses of heat stress given in heat therapy that ultimately promotes the long term health benefits. Heat therapy is not a new idea rather it was practiced since thousands years back in the form of hot bath, sauna bath, steam room. Similarly in Ayurveda there is very comprehensive description of heat therapy in the form of Svedan karma (Sudation therapy). Svedan is a process to induce sweating artificially in a patient who had already undergone Snehan. Svedan is applied for purification of body, as well as in management of various disorders originated due to vitiation of Vata, Kapha Dosha, Meda Dhatu and musculoskeletal disorders. It produces various beneficial effects by augmenting the Agni like clears the channels, liquefies the deposited Dosha, regulates Vata Dosha, helps in removal and pacification of Dosha, augments metabolism (Agni Deepan), increases appetite, flexibility in body parts, softness and shining of skin, removes coldness, stiffness, drowsiness, improves joint motility. However, Svedana karma is vastly used by Ayurveda Physicians in treatment of various disorders but the mechanisms of beneficial effects produced by Svedan Karma are yet not completely explored on scientific basis. In this article, we will discuss and try to establish a possible mechanism of action of Svedana karma in relation to heat stress, mitochondrial adaptation, heat shock protein (HSP) and glucocorticoids as these are secreted under stressful conditions.

A Nutritional Evaluation on Whole Cottonseed Removed Germination Ability by Heat-treatment

  • Hahm, Sahng-Wook;Son, Heyin;Baek, Seong-Gwang;Kwon, Hyeok;Kim, Wook;Oh, Young-Kyoon;Son, Yong-Suk
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.1
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    • pp.39-44
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    • 2013
  • In Korea, wide spread use of whole cottonseed, which is primarily a GMO plant imported from foreign countries and being fed to animals as raw state, has aroused concern that it may disturb the existing ecology of the country unless dispersion of the seed is under proper control. The objective of this study was to elucidate the changes in various nutritive parameters due to heat treatment and to determine the effective condition for removing germination ability of whole cottonseed (WCS). Of the various temperatures applied (76, 78, 80, 85, $100^{\circ}C$/30 min) $85^{\circ}C$ for 30 min was confirmed to be the lowest temperature treatment which resulted in a complete removal of the germination ability of WCS. Therefore, based on the determined temperature condition ($85^{\circ}C$ 30 min) we tried to examine the changes of various nutritional parameters, including nutrient composition, in vitro digestibilities and ruminal protein degradabilities, comparing raw whole cotton seed (RWCS) and heated whole cotton seed (HWCS). Some changes in amino acid composition were observed with heat treatment of WCS, but these were regarded to originate from the variation in plant quality and seed morphology, which are usually affected by different environmental factors during the vegetation period. As for fatty acid composition, no significant differences were observed to occur during heat treatment. However, WCS heated at $85^{\circ}C$ for 30 min in a circulating oven showed a significant decrease (p<0.05) of in situ rumen degradability in both dry matter (DM) and crude protein (CP), as compared to raw WCS. Overall results obtained in the study indicate that the heating condition used in this study, which was proven to be the most appropriate and economic to remove germination ability of WCS, may also improve the nutritional value of the ruminant with regard to reducing its protein degradability within the rumen.