• Title/Summary/Keyword: pH shock

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Effect of hyperthermia on cell viability, amino acid transfer, and milk protein synthesis in bovine mammary epithelial cells

  • Zhou, Jia;Yue, Shuangming;Xue, Benchu;Wang, Zhisheng;Wang, Lizhi;Peng, Quanhui;Hu, Rui;Xue, Bai
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.110-122
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    • 2022
  • The reduction of milk yield caused by heat stress in summer is the main condition restricting the economic benefits of dairy farms. To examine the impact of hyperthermia on bovine mammary epithelial (MAC-T) cells, we incubated the MAC-T cells at thermal-neutral (37℃, CON group) and hyperthermic (42℃, HS group) temperatures for 6 h. Subsequently, the cell viability and apoptotic rate of MAC-T cells, apoptosis-related genes expression, casein and amino acid transporter genes, and the expression of the apoptosis-related proteins were examined. Compared with the CON group, hyperthermia significantly decreased the cell viability (p < 0.05) and elevated the apoptotic rate (p < 0.05) of MAC-T cells. Moreover, the expression of heat shock protein (HSP)70, HSP90B1, Bcl-2-associated X protein (BAX), Caspase-9, and Caspase-3 genes was upregulated (p < 0.05). The expression of HSP70 and BAX (pro-apoptotic) proteins was upregulated (p < 0.05) while that of B-cell lymphoma (BCL)2 (antiapoptotic) protein was downregulated (p < 0.05) by hyperthermia. Decreased mRNA expression of mechanistic target of rapamycin (mTOR) signaling pathway-related genes, amino acid transporter genes (SLC7A5, SLC38A3, SLC38A2, and SLC38A9), and casein genes (CSNS1, CSN2, and CSN3) was found in the heat stress (HS) group (p < 0.05) in contrast with the CON group. These findings illustrated that hyperthermia promoted cell apoptosis and reduced the transport of amino acids into cells, which inhibited the milk proteins synthesis in MAC-T cells.

Transcriptional Analysis Responding to Propanol Stress in Escherichia coli (대장균에서 프로판올 스트레스에 관한 전사분석)

  • Park, Hye-Jin;Lee, Jin-Ho
    • Journal of Life Science
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    • v.22 no.3
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    • pp.417-427
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    • 2012
  • We compared the transcriptome in response to propanol stress in wild-type and propanol-resistant mutant Escherichia coli using the DNA microarray technique. The correlation value of RNA expression between the propanol-treated wild type and the untreated-one was about 0.949, and 50 genes were differentially expressed by more than twofold in both samples. The correlation value of RNA expression between the propanol-treated mutant and the untreated one was about 0.951, and 71 genes in two samples showed differential expression patterns. However, the values between the wild type and mutant, regardless of propanol addition, were 0.974-0.992 and only 1-2 genes were differentially expressed in the two strains. The representative characteristics among differentially expressed genes in W3110 or P19 treated with propanol compared to untreated samples were up-regulation of hest shock response genes and down-regulation of genes relating to ribosome biosynthesis. In addition, many genes were regulated by transcription regulation factors such as ArcA, CRP, FNR, H-NS, GatR, or PurR and overexpressed by sigma factor RpoH. We confirmed that RpoH mediated an important host defense function in propanol stress in E. coli W3110 and P19 by comparison of cell growth rate among the wild type, rpoH disruptant mutant, and rpoH-complemented strain.

Effects of low temperature-adapted Saccharomyces cerevisiae Y297 strain and fermentation temperature on the quality characteristics of Yakju (저온 적응성 효모와 발효온도에 따른 약주의 품질특성 변화)

  • Seo, Dong-Jun;Yeo, Soo-Hwan;Mun, Ji-Young;Baek, Seong Yeol
    • Food Science and Preservation
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    • v.23 no.5
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    • pp.666-672
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    • 2016
  • The objective of this study was to evaluate the effects of low temperature-adapted Saccharomyces cerevisiae Y297 and fermentation temperatures on the quality of Yakju brewed. Physicochemical properties of Yakju brewed were compared pH, total acidity, ethanol, free amino acid, organic acid contents, and volatile flavor compounds in S. cerevisiae Y297 with control treatment. Cooked non-glutinous rice and saccharogenic amylase in koji were mixed with ethanol-producing yeasts and then fermented at $15^{\circ}C$, $20^{\circ}C$, and $25^{\circ}C$ for 20 days. Yakju brewed using the Y297 treatment showed the highest ethanol yield (17.9%) at $20^{\circ}C$. Expression of heat shock protein (HSP) 104 was evaluated by immunoblotting as an indication of adaptation to low temperatures ($15^{\circ}C$); levels of the HSP104 protein were higher in the Y297 treatment than in the control. Organic acid analysis showed that the lactic acid content of Yakju brewed using the control was the highest at $25^{\circ}C$. Finally, free amino acid analysis showed that the Y297 treatment had a higher proportion of essential amino acids than the control. Overall, these results indicate that S. cerevisiae Y297 could be used as a suitable yeast for Yakju brewed under low temperature ($15^{\circ}C$) condition.

Augmentation of Thermotolerance in Primary Skin Fibroblasts from a Transgenic Pig Overexpressing the Porcine HSP70.2

  • Chen, Ming-Yu;Tu, Ching-Fu;Huang, San-Yuan;Lin, Jyh-Hung;Tzang, Bor-Show;Hseu, Tzong-Hsiung;Lee, Wen-Chuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.107-112
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    • 2005
  • A high environmental temperature affects the economic performance of pigs. Heat shock protein 70 (HSP70) has been reported to participate importantly in thermotolerance. This study aims to produce transgenic pigs overexpressing porcine HSP70.2, the highly inducible one of HSP70 members, and to prove the cellular thermotolerance in the primary fibroblasts from the transgenics. A recombinant plasmid in which the sequence that encodes the porcine HSP70.2 gene is fused to green fluorescence protein (GFP) was constructed under the control of cytomegalovirus (CMV) enhancer and promoter. Two transgenic pigs were produced by microinjecting pCMV-HSP70-GFP DNA into the pronucleus of fertilized eggs. Immunoblot assay revealed the varied overexpression level (6.4% and 1.4%) of HSP70-GFP in transgenic pigs. After heating at $45^{\circ}C$ for 3 h, the survival rate (78.1%) of the primary fibroblast cells from the highly expressing transgenic pig exceeded that from the non-transgenic pig (62.9%). This result showed that primary fibroblasts overexpressing HSP70-GFP confer cell thermotolerance. We suggest that transgenic pigs overexpressing HSP70 might improve their thermotolerance in summer and therefore reduce the economic loss in animal production.

Recovery of High-Purity Methane from Piggery Wastewater in the Phase-Separated Anaerobic Process (상분리 혐기성공정에 의한 양돈폐수로부터 고순도 메탄회수)

  • Jung, Jin-Young;Chung, Yun-Chul;Yoo, Chang-Bong
    • 한국신재생에너지학회:학술대회논문집
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    • 2008.05a
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    • pp.210-213
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    • 2008
  • The purpose of this study is to investigate the performances of organic removal and methane recovery in the full scale two-phase anaerobic system. The full scale two-phase anaerobic system was consists of an acidogenic ABR (Anaerobic Baffled Reactor) and a methanognic UASB (Upflow Anaerobic Sludge Blanket) reactor. The volume of acidogenic and methanogenic reactors is designed to 28.3 $m^3$ and 75.3 $m^3$. The two-phase anaerobic system represented 60-82% of COD removal efficiency when the influent COD concentration was in the range of 7,150 to 16,270 mg/L after screening (average concentration is 10,280 mg/L). After steady-state, the effluent COD concentration in the methanogenic reactor showed 2,740 $\pm$ 330 mg/L by representing average COD removal efficiency was 71.4 $\pm$ 8.1% when the operating temperature was in the range of 19-32$^{\circ}C$. The effluent SCOD concentration was in the range of 2,000-3,000 mg/L at the steady state while the volatile fatty concentration was not detected in the effluent. Meanwhile, the COD removal efficiency in the acidogenic reactor showed less than 5%. The acidogenic reactor played key roles to reduce a shock-loading when periodic shock loading was applied and to acidify influent organics. Due to the high concentration of alkalinity and high pH in the effluent of the methanogenic reactor, over 80% of methane in the biogas was produced consistently. More than 70 % of methane was recovered from theoretical methane production of TCOD removed in this research. The produced gas can be directly used as a heat source to increase the reactor temperature.

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몇 가지 PBTs (Persistent, Bioaccumulative, Toxic Chemicals)가 생태계 곤충에 미치는 영향

  • Lee Seun Yeong;Kim Yong Gyun
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2002.11a
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    • pp.123-126
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    • 2002
  • Pollutants that are persistent, bioaccurnulative, and toxic have been linked to numerous adverse effects in human and animals, PBTs include heavy metals, polychlorinated biphenyls (PCBs), dioxins, polycyclic aromatic compounds (PACs) in addition to pesticides. This study focuses on toxic effects of the PBTs except pesticides on insects. Eight PBTs were selected from subgroups: three heavy metals (Pb, Hg, and Cd), two PCB mixtures (Aroclor mixtures 1 and 2), 2,3,7,8-tetrachlorodibenzo-p-dioxin, two monophenols (4-octylphenol and 4-nonylphenol), and tetrabutyltin, Beet armyworm, Spodoptera exigua, was used as test target insect species. Three physiological markers (metamorphosis, immune reaction, and follicle patency) were assessed in each exposure to different doses of the PCBs. Heat-shock proteins as molecular markers were also analyzed in response to the PCBs. All tested PBTs were toxic to metamorphosis from larvae to pupae when they were applied with diet. Two PCB mixtures were the most toxic compounds in this assay by giving significant toxicity at 0.005 ppm, while others had from 10 to 1000 ppm. Dioxin (0.1 ppb), tetrabutyltin (0.1 ppb), Pb (10 ppb), and Hg (0,01 ppb) were potent to inhibit immune reactions analyzed by inducing phenoloxidase activity and blocked phospholipase $A_2$ enzyme, Tetrabutyltin and dioxin significantly induced follicle cell patency, but their effects were lower than that of endogenous juvenile hormone, Dioxin, Pb, Hg, and Cd could induce the expression of heat shock proteins that were detected by immunoblotting against human HSP70 monoclonal antibody. HSP78 and HSP80 were upregulated in response to the PBTs. This expression was detected from the fat body and epidermis at as fast as 4h after injection. All these results clearly suggest that PBTs give significant ecotoxicity to insects that are valuable organisms in our environment.

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Methane Recovery and Performances of Full-scale Two-stage Anaerobic Process Treating Piggery Wastewater (양돈폐수처리시 실규모 이단 혐기성공정의 성능 및 메탄회수)

  • Jung, Jin-young;Chung, Yun-chul;Kang, Shin-hyun;Chung, Hyung-sook
    • Journal of Korean Society on Water Environment
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    • v.21 no.3
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    • pp.256-262
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    • 2005
  • The purpose of this study is to investigate the performances of organic removal and methane recovery by using a full scale two-phase anaerobic system. The full scale two-phase anaerobic process was consists of an acidogenic anaerobic baffled reactor (ABR) and a methanognic upflow anaerobic sludge blanket (UASB) reactor. The volumes of acidogenic and methanogenic reactors were designed to $28.3m^3$ and $75.3m^3$. The two-phase anaerobic system represented 60-82% of COD removal efficiency when the influent COD concentration was in the range of 7,150 to 16,270 mg/L after screening (average concentration is 10,280 mg/L). After steady-state, the effluent COD concentration in the methanogenic reactor showed $2,740{\pm}330 mg/L$ by representing average COD removal efficiency was $71.4{\pm}8.1%$ when the operating temperature was in the range of $19-32^{\circ}C$. The effluent SCOD concentration was in the range of 2,000-3,000 mg/L at the steady state while the volatile fatty acid concentration was not detected in the effluent. Meanwhile, the COD removal efficiency in the acidogenic reactor showed less than 5%. The acidogenic reactor played key roles to reduce a shock-loading when periodic shock loading was applied and to acidify influent organics. Due to the high concentration of alkalinity and high pH in the effluent of the methanogenic reactor, over 80% of methane in the biogas was produced consistently. More than 70% of methane was recovered from theoretical methane production of TCOD removed in this research. The produced gas can be directly used as a heat source to increase the reactor temperature.

The Molecular Biomarker Genes Expressions of Rearing Species Chironomus riparious and Field Species Chironomus plumosus Exposure to Heavy Metals (실내종 Chironomus riparious와 야외종 Chironomus plumosus의 중금속 노출에 따른 분자지표 유전자 발현)

  • Kim, Won-Seok;Kim, Rosa;Park, Kiyun;Chamilani, Nikapitiya;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.48 no.2
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    • pp.86-94
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    • 2015
  • Chironomous is aquatic insect belonging to order Diptera, family Chironomidae. Their larval stage can be found mainly in aquatic benthic environment, hence good model organism to study environmental toxicology assessments and consider as useful bio indicators of contamination of the aquatic environment. In this study, Chironomus Heat Shock Proteins, Cytochrome 450, Glutathione S-transferase, Serine-type endopeptidase gene expressions were compared between polluted field areas (Chironomus plumosus) and under laboratory conditions (Chironomus riparious) to investigate molecular indicators for environmental contaminant stress assessment. Heavy metal (Al, Fe, Mn, Cu, Cr, Zn, Se, Pb, As, Cd) concentrations in sediments collected from three study areas exceeded the reference values. Moreover, HSPs, CYP450 and GST gene expression except SP for C. plumosus showed higher expression than C. riparious gene expression. Similar gene expression pattern was observed in C. riparious that exposed environment waters up to 96 h when compared to C. plumosus exposed to waters that grown in lab conditions. In summary, this comparative gene expression analysis in Chironomous between field and laboratory condition gave useful information to select candidate molecular indicators in heavy metal contaminations in the environment.

Identification of Muscle Proteins Related to Objective Meat Quality in Korean Native Black Pig

  • Hwang, I.H.;Park, B.Y.;Cho, S.H.;Kim, J.H.;Choi, Y.S.;Lee, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.11
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    • pp.1599-1607
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    • 2004
  • This study examined the effects of pre-slaughter fasting, chasing stress and chiller ageing on objective meat quality, and their relations to the proteome profile of longissimus muscle using 20 male Korean native black pigs. Treatments were composed of two levels of pre-slaughter feed withdrawal, two levels of pre-slaughter stress and four chiller ageing times. A 15 min chasing stress immediately prior to slaughter significantly (p<0.05) decreased detectable levels of $\mu$-calpain activity during rigor development and chiller ageing, but did not have any direct effect on objective meat quality. On the other hand, pigs fed until the morning of slaughter resulted in significantly (p<0.05) higher hunter L* value and cooking loss than those which received an 18 h feed withdrawal prior to slaughter. Cooking loss and hunter L* value were constant during 7 d of chiller ageing, followed by significant increases at 14 d. The fed animals showed a significantly (p<0.05) higher hunter a* value at both 3 and 7 d, while the other group maintained a stable redness for 7 d. WB-shear force was not affected by the pre-slaughter treatments, but had significant (p<0.05) linear reduction from 1 to 7 d. A gelbased proteome analysis was performed on selected animals for low and high hunter L* values at 1 d. Ten and five spots had greater than two-fold spot densities for the low and high hunter L* groups, respectively. The ten spots included chain A, deoxyribounclease I complex with actin, heat shock protein 27 kDa, a protein similar to cardiac $Ca^{2+}$ release channel, and myosin heavy chain, while the five spots included chain A aldehyde dehydrogenase, glycerol-3 phosphate dehydrogenase, and hemoglobin alpha chain. In general, feeding until the morning of slaughter resulted in more desirable meat color, but appeared to reduce palatability due to increased cooking loss. Proteome analysis demonstrated that various proteins were concomitantly involved in the determination of final meat color. The most noticeable observation in the current study was that various isoforms for a particular protein differed in degradation and/or expression rate depending on meat quality.

Neuroprotective Effects of Pinelliae Rhizoma Water-Extract by Suppression of Reactive Oxygen Species and Mitochondrial Membrane Potential Loss in a Hypoxic Model of Cultured Rat Cortical Cells. (배양대뇌신경세포 저산소증모델에서 유해산소생성억제 및 사립체막전위 소실방지에 의한 반하(半夏)의 신경세포사 억제 효능)

  • Kwon, Gun-Rok;Moon, Il-Soo;Lee, Won-Chul
    • Journal of Life Science
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    • v.19 no.5
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    • pp.598-606
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    • 2009
  • Oxidative stress by free radicals is a major cause of neuronal cell death. Excitotoxicity in hypoxia/ischemia causes an increase in reactive oxygen species (ROS) and a loss of mitochondrial membrane potential (MMP), resulting in dysfunction of the mitochondria and cell death. Pinelliae Rhizoma (PR) is a traditional medicine for incipient stroke. We investigated the effects of PR Water-Extract on the modulation of ROS and MMP in a hypoxic model using cultured rat cortical cells. PR Water-Extract was added to the culture medium at various concentrations (0.25${\sim}$5, 5.0 ${\mu}g/ml$) on day in vitro 12(DIV12), given a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 3 hr), and cell viability was assessed on DIV15 by Lactate Dehydrogenase Assay (LDH assays). PR Water-Extract showed a statistically significant effect on neuroprotection (10${\sim}$15% increase in viability; p<0.01) at 1.0 and 2.5 ${\mu}g/ml$ in normoxia and hypoxia. Measurement of ROS production by $H_2DCF-DA$ stainings showed that PR Water-Extract efficiently reduced the number and intensity of ROS-producing neurons, especially at 1 hr post shock and DIV15. When MMP was measured by JC-1 stainings, PR Water-Extract efficiently maintained high-energy charged mitochondria. These results indicate that PR Water-Extract protects neurons in hypoxia by preventing ROS production and preserving the cellular energy level.