• Title/Summary/Keyword: pH shock

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Studies on the Properties of Charolais Semen (Charolais의 정액 성상에 관한 연구)

  • 고광두;손봉환;변명대;김선환
    • Korean Journal of Animal Reproduction
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    • v.4 no.1
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    • pp.20-27
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    • 1980
  • This experiment was to study semen properties of Charolais for the a, pp.ication ot artificial insemination. The result obtained were summarized as follows: 1. In the preservation of liquid semen for 6 days, the survival rates of Charolais semen averaged 57.14% in skim milk solution and 58.17% in tris buffer solution. There were not differences. 2. Recovery of semen after thawing was vigorous in the semen that was diluted and frozen in 48 hrs. 3. The real rates of survival sperm for Charolais averaged 83% after living sperm was diluted and stained for 6 days. 4. Methylene blue reduction test diluted semen was fresh when it was diluted within 48 hrs. 5. If the diluted semen was preserved below 5$^{\circ}C$ in Charolais, the pH decreased by 0.2 in a day. 6. Diluted semen was more resistant to the cold shock than fresh semen. 7. In resistance against hot shock, sperm was almost dead in 20 minutes in 46.5$^{\circ}C$ in diluted semen, while it was dead in 30 minutes in 42.5$^{\circ}C$ in diluted semen. 8. In examination of morphological changes of sperm acrosome for 6 days, normal sperm in skim milk solution and tris buffer solution was 80% and 76.97% respectively, swelling sperm 12.8% and 15.27%, deficient sperm 0.6% and 0.97% abnormal staining 3.07% and 5.25%, immature sperm 0.28%, and 0.23%, whereas other abnormal sperm was 1.28% and 1.42%.

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Effect of Pretreatment Method on Lipid Extraction from Enteromorpha intestinalis (해조류 파래로부터 지질 추출에 미치는 전처리 방법의 영향)

  • Jeong, Gwi-Taek;Park, Don-Hee
    • KSBB Journal
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    • v.29 no.1
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    • pp.22-28
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    • 2014
  • In this study, we investigate the effect of pretreatment method on lipid extraction from Enteromorpha intestinalis using physical, thermo-chemical, and enzymatic process such as ultrasonication, high temperature treatment, freezing, microwave irradiation, osmotic shock, pH shock, homogenizing, and enzymatic treatment. In pretreatment with separated lipid extraction, the high extraction yield was obtained by high temperature treatment ($121^{\circ}C$ for 5 min) with 0.1 N HCl, which is 1.4 times higher than that of control. In pretreatment with direct lipid extraction, the high extraction yields were obtained by 0.1 N HCl pretreatment, microwave irradiation (700W, 1 min with twice), and 10% NaCl pretreatment, which is 1.45 times higher than that of control. In the result of enzymatic pretreatment with 17 kinds of enzymes, Cellic CTec II showed the high extraction yield of 5.3%, and which is 1.9 times higher than that of control. Moreover, the extraction yield was increased by the increase of enzyme amounts. In 10% enzyme amount, about 5.8% yield was obtained.

Overproduction and Purification of Ribose-Binding Proteins from the Wild-Type Mutant and Revertant Strains in Escherichia coli (리보스 결합단백질의 대량생산을 위한 야생형 수송결합변이, 복귀변이 유전자의 클로닝과 이들 단백질의 순수정제)

  • ;Randall Linda L.
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.291-297
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    • 1988
  • Three alleles of rbsB gene, rbsB, rbsB103, and rbsB106 from the wild type, the mutant and the revertant strain, respectively, were cloned for overproduction of proteins under the control of lambda $P_{L}$ promoter. Five different species of precursor and mature ribose-binding proteins were purified to homogeneity using DEAE-Sephadex column chromatography, osmotic shock pocedure, CM-Sephadex column chromatography, and Chromatofocusing column chromaography. pI of the precursor proteins and mature proteins were determined and found to be pH 8.0 and 7.5, respectively. The purified proteins were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing.

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Increased Viability of Sub-lethal Heat Shocked Salmonella Typhimurium on Acids and Oxidants (열충격 Salmonella Typhimurium의 산과 산화제에서 생존력 증가)

  • Moon, Bo-Youn;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.712-716
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    • 2008
  • In an effort to evaluate Salmonella food safety using combinations of preservation techniques, its viabilities when exposed to HCl, acetic acid, and the oxidative agents (hydrogen peroxide and butyl hydrogen peroxide), were analyzed using sub-lethal heat-shocked Salmonella Typhimurium at $56^{\circ}C$. 2D gel electrophoresis and MALDI-TOF MS analyses were also conducted to determine the expression and repression of proteins in heat-shocked cells. Heat-shocked S. Typhimurium evidenced a reduction of viable counts by 1-2 log CFU/mL. However, viality of non heat-shocked S. Typhimurium decreased markedly by 5-6 log CFU/mL at a pH 4 in response to acid and oxidative stresses. Sub-lethal heat treatment greatly increased the resistance of S. Typhimurium against acid and oxidant agents. As for 2D gel electrophoresis and protein identification via MALDI-TOF MS, 17 major proteins in non heat-shocked S. Typhimurium were detected, and only 13 proteins among these proteins were detected in heat-shocked S. Typhimurium. The heat shock proteins such as DnaK and small heat shock proteins were included, and may be associated with the resistance of S. typhimurium against exposure to acids and oxidants. Therefore, even though the promising hurdle technology using the combined mild treatments including heat was applied to S. Typhimurium, the proper heat treatment to reduce its crossprotection activity toward the following preservative agents might be considered.

Expression of Heat Shock Protein HspA2 in Human Tissues (인간 조직에서 Heat Shock Protein A2 (HspA2) 단백질의 발현)

  • Son, W.Y.;Hwang, S.H.;Han, C.T.;Lee, J.H.;Choi, Y.J.;Kim, S.;Kim, Y.C.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.225-230
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    • 1999
  • In mouse, the heat shock protein 70-2 (hsp70-2) is found to have special function in spermatogenesis. Based on the observation, the hypothesis that human hspA2 (human gene; 98.2% amino acid homology with hsp70-2) might have important function in spermatogenesis in human testes was proposed. To test the hypothesis, we examined the expression of hspA2 in human tissues. Expression vector pDMC4 for expression of the human hspA2 protein using pTricHisB (invitrogen, USA) was constructed and the expressed hspA2 protein was cross-reacted with antiserum 2A raised against mouse hsp70-2 protein. Based on the cross-reactivity, we determined the expression level of hspA2 protein in human tissues by western blot analysis using the antiserum 2A. We demonstrated that antiserum 2A antibodies detected human hspA2 protein with specificity which was produced in the E.coli expression system. On Western blot analyses, significant hspA2 expression was observed in testes with normal spermatogenesis, whereas a low level of hspA2 was expressed in testis with Sertoli-cell only syndrome. Also, a small amount of hspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. These results demonstrate that the hspA2 protein is highly expressed in male specific germ cells, which in turn suggests that hspA2 protein might playa specific role during meiosis in human testes as suggested in the murine model. However, further studies should be attempted to determine the function of hspA2 protein in human spermatogenesis.

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Studies on the Biological Action of Hyperthermia on Tumor Cell Mortality (1) (腫瘍細胞가 致死에 미치는 溫熱處理의 生物學的 作用에 관한 硏究(1))

  • Kang, Man-Sik;Lee, Chung-Choo
    • The Korean Journal of Zoology
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    • v.26 no.2
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    • pp.69-81
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    • 1983
  • The biological effect of hyperthermia on the SCK tumor cells in vitro were analyzed in several respects. A comparision of the survival curve of SCK tumor cells in vitro and in vivo following hyperthermia demonstrated that the cytocidal effect of heating is far greater on the cells in vivo than on the cells in vitro. The pH change in the SCK tumor upon being heated at $43.5^\\circC$ started out at 7.05 and increased to 7.18 during the first 7 min of heating and then rapidly declined to 6.67 by 30 min. Contrary to the decrease in pH in the heated tumors, the pH in the muscle increased significantly when heated to $43.5-45.0^\\circC$. Following hyperthermia at $43.5^\\circC$ for 30 min, a maximum increase in the lactic acid content in the tumor and liver was observed at 1 hr and 3 hr, respectively. The increase in the tumor was followed by a gradual decrease below the control level, whereas the increase in the liver was maintained at quite a steady level for 24 hr. The hyperthermia at $43.5^\\circC$ for 1 hr exhibited a general tendency that high molecular proteins decrease markedly, whereas most of low molecular proteins increase. The most prominent change was that the heat shock protein 70K increased significantly along with other low molecular proteins in heat shocked cells.

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Purification and Structural Characterization of Cold Shock Protein from Listeria monocytogenes

  • Lee, Ju-Ho;Jeong, Ki-Woong;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.33 no.8
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    • pp.2508-2512
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    • 2012
  • Cold shock proteins (CSPs) are a family of proteins induced at low temperatures. CSPs bind to single-stranded nucleic acids through the ribonucleoprotein 1 and 2 (RNP 1 and 2) binding motifs. CSPs play an essential role in cold adaptation by regulating transcription and translation via molecular chaperones. The solution nuclear magnetic resonance (NMR) or X-ray crystal structures of several CSPs from various microorganisms have been determined, but structural characteristics of psychrophilic CSPs have not been studied. Therefore, we optimized the purification process to obtain highly pure Lm-Csp and determined the three-dimensional structure model of Lm-Csp by comparative homology modeling using MODELLER on the basis of the solution NMR structure of Bs-CspB. Lm-Csp consists of a ${\beta}$-barrel structure, which includes antiparallel ${\beta}$ strands (G4-N10, F15-I18, V26-H29, A46-D50, and P58-Q64). The template protein, Bs-CspB, shares a similar ${\beta}$ sheet structure and an identical chain fold to Lm-Csp. However, the sheets in Lm-Csp were much shorter than those of Bs-CspB. The Lm-Csp side chains, E2 and R20 form a salt bridge, thus, stabilizing the Lm-Csp structure. To evaluate the contribution of this ionic interaction as well as that of the hydrophobic patch on protein stability, we investigated the secondary structures of wild type and mutant protein (W8, F15, and R20) of Lm-Csp using circular dichroism (CD) spectroscopy. The results showed that solvent-exposed aromatic side chains as well as residues participating in ionic interactions are very important for structural stability. Further studies on the three-dimensional structure and dynamics of Lm-Csp using NMR spectroscopy are required.

Charaterization of GroEL Homolog from Streptococcus pneumoniae (폐렴구균 열충격 단백질 GroEL의 특성)

  • Choi, In hwa;Kim, Su Nam;Kim, Seung Hwan;Kweon, Chang Hee;Pyo, Suhk Neung;Rhee, Dong Kwon
    • Korean Journal of Microbiology
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    • v.34 no.1_2
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    • pp.43-50
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    • 1998
  • Induction and purification of the GroEL homolog from Streptococcus pneumolliae were characterized. The stress conditions were determined by temperature, ethanol, NaCI, $H_2O_2$ methyl methanesulfonate, and ethyl methanesulfonate. And stress induced proteins were analyzed using [$^{35}S$]-methionine labeling method. Heat shock induced the synthesis of a set of about 3 heat shock proteins (hsps) (65, 73, and 84-kDa). Of those 3 hsps, a 65 kDa protein, hsp65, was purified by DEAE-Sephacel and ATP-agarose column chromatography, and used for antibody preparation. Immunoblot analysis employing antisera raised against pneumococcus hsp65 demonstrated cross-reactivity with a 60 kDa protein in Eschericha coli. Also cross reaction of the purified p65 with anti-Escherichia coli GroEL monoclonal antibody demonstrated that pneumococcal hsp65 is the GroEL homolog.

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Overexpression of the Small Heat Shock Protein, PtsHSP19.3 from Marine Red Algae, Pyropia tenera (Bangiales, Rhodophyta) Enhances Abiotic Stress Tolerance in Chlamydomonas

  • Jin, Yujin;Yang, Sungwhan;Im, Sungoh;Jeong, Won-Joong;Park, EunJeong;Choi, Dong-Woog
    • Journal of Plant Biotechnology
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    • v.44 no.3
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    • pp.287-295
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    • 2017
  • Water temperature is one of the major factors that impacts the growth and life cycle of Pyropia tenera, one of the most valuable and cultivated marine red algae belonging to Bangiales (Rhodophytes). We analyzed transcriptome from gametophyte of P. tenera under normal and high temperature conditions, and identified four small heat shock proteins (sHSPs). They have no significant amino acid sequence homology with known proteins in public databases except PhsHSP22 from Pyropia haitanensis. PtsHSP19.3 gene responded to high temperature but slightly or not to desiccation, freezing or high salt condition. When the PtsHSP19.3 gene was overexpressed in Chlamydomonas reinhardtii, transformed Chlamydomonas lines revealed much higher growth rate than that of control cells under heat stress condition. Transformed cells also grew well in those of the control cell onto the medium containing high salt or $H_2O_2$. When the PtsHSP19.3 was fused to GFP and introduced into tobacco protoplast, fluorescence was detected at several spots. Results indicate that PtsHSP19.3 may form super-molecular assembles and be involved in tolerance to heat stress.

Disinfection of Water by Ultrasonic Irradiation (초음파 검사에 의한 수중의 살균처리)

  • 손종렬;유병성
    • Journal of environmental and Sanitary engineering
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    • v.14 no.1
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    • pp.54-61
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    • 1999
  • The industrial techniques of ultrasound have been used in the various fields, such as cleaning, medical surgery, emulsification, cell disruption etc. Especially the application of cell disruption was interested in the field of disinfection process in water by ultrasonic irradiation. It has been recognized that the ultrasounds are irradiated in aqueous solution, cavitation bubbles are generated and shock waves of high temperature and pressure are emitted as the bubbles are developed and finally broken, which function as a energy source to promote reaction efficiencies of various kinds of chemical reactions such as disinfection reaction in water. Therefore, this study was performed to apply the ultrasound for the disinfection method of infected drinking raw water and to discuss the limiting factors such as pH, sample volume and reaction temperature influenced on the removal efficiency of E. coli from experimental analysis of the results obtained in bench-scale plant. For the experiments to measure the influence of reaction parameters in the ultrasonic disinfection process, escalated reactivity of aqueous solutions was excellent when pH in aqueous solution was low, and sample volume was small. On the contrary, the reactivity of disinfection became elevated when reaction temperature was high. It was found that the rate constant of disinfection reaction was applied by Chick's law, reaction kinetics of Chick's law was irreversible and pseudo-first order at all the tested conditions.As a conclusion it appeared that the technology using ultrasonic irradiation can be applied to the treatment of disinfection in infected water which are difficult to be treated by conventional methods.

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