• Title/Summary/Keyword: Hsp 70

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Gene Expression Analyses of Mutant Flammulina velutipes (Enokitake Mushroom) with Clogging Phenomenon

  • Ju-Ri Woo;Doo-Ho Choi;Muhammed Taofiq Hamza;Kyung-Oh Doh;Chang-Yoon Lee;Yeon-Sik Choo;Sangman Lee;Jong-Guk Kim;Heeyoun Bunch;Young-Bae Seu
    • Mycobiology
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    • v.50 no.5
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    • pp.366-373
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    • 2022
  • Regulation of proper gene expression is important for cellular and organismal survival, maintenance, and growth. Abnormal gene expression, even for a single critical gene, can thwart cellular integrity and normal physiology to cause diseases, aging, and death. Therefore, gene expression profiling serves as a powerful tool to understand the pathology of diseases and to cure them. In this study, the difference in gene expression in Flammulina velutipes was compared between the wild type (WT) mushroom and the mutant one with clogging phenomenon. Differentially expressed transcripts were screened to identify the candidate genes responsible for the mutant phenotype using the DNA microarray analysis. A total of 88 genes including 60 upregulated and 28 downregulated genes were validated using the real-time quantitative PCR analysis. In addition, proteomic differences between the WT and mutant mushroom were analyzed using two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF). Interestingly, the genes identified by these genomic and proteomic analyses were involved in stress response, translation, and energy/sugar metabolism, including HSP70, elongation factor 2, and pyruvate kinase. Together, our data suggest that the aberrant expression of these genes attributes to the mutant clogging phenotype. We propose that these genes can be targeted to foster normal growth in F. velutipes.

Suboptimal Mitochondrial Activity Facilitates Nuclear Heat Shock Responses for Proteostasis and Genome Stability

  • Dongkeun Park;Youngim Yu;Ji-hyung Kim;Jongbin Lee;Jongmin Park;Kido Hong;Jeong-Kon Seo;Chunghun Lim;Kyung-Tai Min
    • Molecules and Cells
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    • v.46 no.6
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    • pp.374-386
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    • 2023
  • Thermal stress induces dynamic changes in nuclear proteins and relevant physiology as a part of the heat shock response (HSR). However, how the nuclear HSR is fine-tuned for cellular homeostasis remains elusive. Here, we show that mitochondrial activity plays an important role in nuclear proteostasis and genome stability through two distinct HSR pathways. Mitochondrial ribosomal protein (MRP) depletion enhanced the nucleolar granule formation of HSP70 and ubiquitin during HSR while facilitating the recovery of damaged nuclear proteins and impaired nucleocytoplasmic transport. Treatment of the mitochondrial proton gradient uncoupler masked MRP-depletion effects, implicating oxidative phosphorylation in these nuclear HSRs. On the other hand, MRP depletion and a reactive oxygen species (ROS) scavenger non-additively decreased mitochondrial ROS generation during HSR, thereby protecting the nuclear genome from DNA damage. These results suggest that suboptimal mitochondrial activity sustains nuclear homeostasis under cellular stress, providing plausible evidence for optimal endosymbiotic evolution via mitochondria-to-nuclear communication.

Dietary Exogenous α-Amylase Modulates the Nutrient Digestibility, Digestive Enzyme Activity, Growth-Related Gene Expression, and Diet Degradation Rate of Olive Flounder (Paralichthys olivaceus)

  • Md. Tawheed Hasan;Hyeon Jong Kim;Sang-Woo Hur;Seong-Mok Jeong;Kang-Woong Kim;Seunghan Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1390-1401
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    • 2023
  • In this study, a 12-week feeding experiment was conducted to characterize the effects of exogenous α-amylase on the growth, feed utilization, digestibility, plasma α-amylase activity, feed degradation rate, and fecal particle size of olive flounder (Paralichthys olivaceus). Diet was supplemented with 0 (AA0; control), 100 (AA100), 200 (AA200), or 400 (AA400) mg/kg of α-amylase, respectively. Fish (273.1 ± 2.3 g) were stocked into 12 tanks (25 fish/1,000-L tank) and 3 tanks were randomly selected for each diet group. As a result, α-amylase was found to have no significant effects (p ≥ 0.05) on the growth, feed utilization parameters, and whole-body proximate compositions. α-Amylase-treated fish exhibited only a significant increase in the apparent digestibility coefficient of carbohydrates compared to the controls. In addition, in vitro analyses revealed that α-amylase dose-dependently increased (p < 0.05) the feed degradation rate, while photographs of the intestinal content after 2, 4, and 8 h of feeding demonstrated an improved degradation rate in the α-amylase-treated groups. Plasma α-amylase content was higher in the AA200 and AA400 groups, whereas the control group produced significantly larger-sized fecal particles (90% size class) than these two groups. In the intestine, no changes were observed in the expression levels of the immune-related TNF-α, IL-1β, IL-2, immunoglobulin-M, HSP-70, lysozyme, and amylase alpha-2A. However, growth-related genes IGF-1, IGF-2, TGF-β3, and growth hormone genes were upregulated in muscle tissues. Collectively, exogenous α-amylase has positive roles in the modulation of the digestibility coefficient, blood α-amylase concentration, growth-related gene expression, and diet degradation for improved digestion in olive flounder.

Evaluation of Construction Operation and Design Properties of CLSM for Corrugated Pipe in Underground (파형강관을 이용한 지하매설물용 뒤채움재 설계 및 시공성 평가)

  • Lee Kwan-Ho;Park Jae-Heon
    • International Journal of Highway Engineering
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    • v.8 no.2 s.28
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    • pp.63-74
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    • 2006
  • During the construction of circular underground pipe, the non-proper compaction along the pipe and the decrease of compaction efficiency have been the main problems to induce the failure of underground pipe or facility. The use of CLSM (controlled low strength materials) should be one of the possible applications to overcome those problems. In this research, the full-scaled field test and the numeric analysis using PENTAGON-3D FEM program were carried out for three different cases on the change of backfill materials, including the common sand, the soil from construction site, and the CLSM. From the full-scaled test in field, the use of in-situ CLSM as backfill materials reduced the vertical and lateral deformation of the pipe, as well as the deformation of the ground surface. The main reason for reducing the deformation would be the characteristics of the CLSM, especially self-leveling and self-hardening properties. The measured earth pressure at the surround of the corrugated pipe using the CLSM backfills was the smaller than the other cases, and the absolute value was almost zero. Judging from the full-scaled field test and FEM analysis, the use of CLSM as backfill materials should be one of the best choices reducing the failure of the underground pipes.

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Effect of Heat-Killed Enterococcus faecalis, EF-2001 on C2C12 Myoblast Damage Induced by Oxidative Stress and Muscle Volume Decreased by Sciatic Denervation in C57BL/6 Mice (산화스트레스에 의해 유도된 C2C12 근세포 손상과, 신경절제에 의해 근감소가 유도된 C57BL/6 마우스에서 열처리 사균체 엔테로코커스 패칼리스 EF-2001의 효과)

  • Chang, Sang-Jin;Lee, Myung-Hun;Kim, Wan-Joong;Chae, Yuri;Iwasa, Masahiro;Han, Kwon-Il;Kim, Wan-Jae;Kim, Tack-Joong
    • Journal of Life Science
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    • v.29 no.2
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    • pp.215-222
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    • 2019
  • Muscle dysfunction may arise from skeletal muscle atrophy caused by aging, injury, oxidative stress, and hereditary disease. Powdered heat-killed Enterococcus faecalis (EF-2001) has anti-allergy, anti-inflammatory, and anti-tumor effects. However, its antioxidant and anti-atrophy effects are poorly characterized. In this study, we examined the effects of EF-2001 on muscle atrophy. To determine the protective effect of EF-2001 on oxidative stress, C2C12 myoblasts were treated with $H_2O_2$ to induce oxidative stress. This induced cell damage, which was reduced by treatment with EF-2001. The mechanism of EF-2001's effect was examined in response to oxidative stress. Treatment with EF-2001 reversed the expression of HSP70 and SOD1 proteins. Also, mRNA levels of Atrogin-1/MAFbx and MuRF1 increased under oxidative stress conditions but decreased following EF-2001 treatment. To evaluate muscle volume, two and three dimensional models of the muscles were analyzed using micro-CT. As expected, muscle volume decreased after sciatic denervation and recovered after oral administration of EF-2001. Therefore, EF-2001 is a candidate for the treatment of muscular atrophy, and future discovery of the additional effects of EF-2001 may yield further applications as a functional food with useful activities in various fields.

Reduction effects of N-acetyl-L-cysteine, L-glutathione, and indole-3-acetic acid on phytotoxicity generated by methyl bromide fumigation- in a model plant Arabidopsis thaliana (모델식물 애기장대에 대한 훈증제 메틸브로마이드의 약해발생 및 N-acetyl-L-cysteine, L-glutathione, indole-3-acetic acid의 약해억제 효과)

  • Kim, Kyeongnam;Kim, Chaeeun;Park, Jungeun;Yoo, Jinsung;Kim, Woosung;Jeon, Hwang-Ju;Kim, Jun-Ran;Lee, Sung-Eun
    • Korean Journal of Environmental Biology
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    • v.39 no.3
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    • pp.354-361
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    • 2021
  • Understanding the phytotoxic mechanism of methyl bromide (MB), an essential fumigant during the quarantine and pre-shipment process, is urgently needed to ensure its proper use and reduce international economic losses. In a previous study, two main MB-induced toxic mechanisms such as reactive oxygen species (ROS) and auxin distribution were selected by analyzing transcriptomic analysis. In the study, a 3-week-old A. thaliana was supplied with 1 mM ROS scavengers [N-acetyl-L-cysteine (NAC) or L-glutathione (GSH)] and 1µM indole-3-acetic acid(IAA) three times every 12 h, and visual and gene expression assessments were performed to evaluate the reduction in phytotoxicity by supplements. Phytotoxic effects on the MB-4h exposed group were decreased with GSH application compared to the other single supplements and a combination of supplements at 7 days post fumigation. Among these supplements, GSH at a concentration of 1, 2, and 5mM was suppled to A. thaliana with MB-fumigation. During a long-term observation of 2 weeks after the fumigation, 5 mM GSH application was the most effective in minimizing MB-induced phytotoxic effects with up-regulation of HSP70 expression and increase in main stem length. These results indicated that ROS was a main key factor of MB-induced phytotoxicity and that GSH can be used as a supplement to reduce the phytotoxicity of MB.

The Effect of Heat Co-treatment on Acute Lung Injury of the Rat Induced by Intratracheal Lipopolysaccharide (내독소 투여 직후 가해진 열충격이 백서의 급성폐손상에 미치는 영향)

  • Na, Joo Ock;Shim, Tae Sun;Lim, Chae-Man;Lee, Sang Do;Kim, Woo Sung;Kim, Dong Soon;Kim, Won Dong;Koh, Younsuck
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.4
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    • pp.355-366
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    • 2002
  • Background : The heat shock protein (HSP) 70 families are known to protect cells against the irreversible tissue injury induced by stress and to induce the recovery of cell function during stress. Heat pretreatment was reported to decrease the acute lung injury (ALI) of rats induced by lipopolysaccharide (LPS). However, the role of heat shock with LPS co-treatmenton ALI is unclear. The purpose of this study was to investigate the effect of heat treatment, which was given immediately after the beginning of ALI induced by LPS intratracheally administered in rats. Methods : Either saline (saline group) or LPS was intratracheally instilled without heat treatment (LPS group). In addition, heat was conducted 18 hours prior to the instillation of LPS (pre-treatment group) and conducted immediately after instillation of LPS (co-treatment group). Six hours after the LPS or saline treatment, blood, bronchoalveolar lavage (BAL) fluid and lung tissue samples were obtained. The myeloperoxidase (MPO) activity and the heat shock protein expression in the lung tissue, the differential counts of the polymorphonuclear leukocytes (PMN) in the BAL fluids, and the LDH, protein, $IL-1{\beta}$, $TNF-{\alpha}$ and IL-10 levels in BAL fluid and serum were measured. Results : 1) The MPO activity, the differential PMN counts in the BAL fluid, BAL fluid and serum cytokines were higher in the LPS, the heat pre-treatment and co-treatment group than those of the saline group (p value <0.05). 2) The MPO activity and the protein level in the BAL fluid from the heat co-treatment group were similar to those of the LPS group. 3) The serum $TNF-{\alpha}$ level of the heat co-treatment group was significantly higher than that of the LPS group (p=0.01). Conclusion : Heat shock response administered immediately after a LPS instillation did not attenuate the ALI in this model.

Characteristics of Soybean Soaking Water after Heat Treatment (대두 열처리 담금 용액의 특성)

  • Yoon, Hye-Hyun;Jeon, Eun-Jae
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1098-1103
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    • 2003
  • Soybeans released proteins when immersed in water at $50{\sim}60^{\sim}C$. We investigated the changes in the characteristics of soybean when soaked in water at different temperatures and studied the electrophoretic properties of soy proteins in recommended Korean soybean varieties after heat treatment. Soybean seeds were heated in soaking water at temperatures of 30, 40, 50, 60, $70^{\circ}C$ for 90 min, and also from 10 to 150min at $60^{\circ}C$. The pH value of the water decreased with heating time at $60^{\circ}C$, and the amount of soluble solids increased with temperature and heating time. The protein concentration of the solution increased with temperature and time. From SDS-PAGE of the proteins in soaking water, we detected two new bands of 16 kDa- and 31 kDa-proteins from the Korean soybean varieties on heat treatment.

Effect of Dietary Inclusion of Various Sources of Green Tea on Immune System and Challenging Test of Juvenile Olive Flounder Paralichthys olivaceus (사료내 녹차 첨가가 넙치 유어기의 면역성 및 세균 공격성에 미치는 영향)

  • Cho Sung-Hwoan;Lee Sang-Mok;Park Byum-Hee;Ji Sung-Choon;Kwon Mun-Gyeong;Kim Yi-Cheong;Lee Jong-Ha;Park Sagn-Eun;Han Hyoung-Kyun
    • Journal of Aquaculture
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    • v.19 no.2
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    • pp.84-89
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    • 2006
  • Effect of dietary inclusion of various sources of green tea on growth, immune system and challenging test of juvenile olive flounder Paralichthys olivaceus was investigated. Five experimental diets with triplicates were prepared: control, raw leaves, dry leaves, by-product and extract. Twenty five (an initial body weight of 52.5 g) were randomly distributed into 15 of 180 L flow-through tanks. Nutrient requirements of the experimental diets satisfied growth of juvenile olive flounder. The feeding trial lasted for 7 weeks. After 7-week feeding trial, blood were sampled from three randomly chosen fish for serum analysis of Iysozyme and bactericidal activity, and ten fish were infected with Edwardsiella tarda for challenging test from each tank. Weight gain (g/fish) of fish fed the diet containing extract and control diet was significantly higher than that of fish fed the other diets. Feed efficiency ratio for fish fed the diet containing extract and control diet was significantly higher than that for fish fed the diets containing raw leaves and by-product, but not significantly different from that for fish fed the diet containing dry leaves. Serum Iysozyme activity (units/ml) of fish fed the diets containing dry leaves and extract was significantly higher than that of fish fed the diets containing raw leaves and by-product, but not significantly different from that of fish fed the control diet. Serum bactericidal activity (${\times}10^6$ bacteria/ml) of fish fed the diet containing dry leaves and extract was significantly lower than that of fish fed the diets containing raw leaves, by-product and control diet in 3 hour. However, serum bactericidal activity of fish fed the diet containing extract was significantly lower than that of fish fed the other diets in 6 hour. And serum bacterial activity was low in fish fed the diets containing dry and raw leaves, by-product, and control in 6 hour in order. Accumulative mortality (%) of fish fed the control diet was low compared to that of fish fed the diets containing raw leaves and by-product, but high compared to that of fish fed the diets containing dry leaves and extract although no significant difference was found among treatments. In considering above results, dietary inclusion of extract and dry leaves of green tea seemed to be highly effective to improve immune system and endurance against E. tarda infection of juvenile olive flounder.

Effects of Short-term Acute Heat Stress on Physiological Responses and Heat Shock Proteins of Hanwoo Steer (Korean Cattle)

  • Baek, Youl-Chang;Kim, Minseok;Jeong, Jin-Young;Oh, Young-Kyoon;Lee, Sung-Dae;Lee, Yoo-Kyung;Ji, Sang-Yun;Choi, Hyuck
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.173-182
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    • 2019
  • This study was performed to evaluate the effect of heat stress on the status of physiological responses, blood parameter, serum T3 and cortisol, and heat shock proteins (HSP 27, 70, and 90) of Hanwoo cattle. Six Hanwoo steers (242.8 ± 7.2 kg of BW) were housed in the climate-controlled respiration chambers. The experiment consisted of 7 days (control; 0 day) at thermoneutral (air temperature (Ta) of 15℃ and relative humidity (RH) of 60%; temperature-humidity index (THI) = 64), and by 3 and 6 days (treatment groups) at heat stress (Ta of 35℃ and RH of 60%; THI = 87). Body temperature of each parts (frank, rump, perineum and foot) and rectal temperature elevated in heat stress groups (3 days and 6 days) than the control group (0 day). Respiration rates increased in 3 days and 6 days (88.5 ± 0.96 bpm and 86.3 ± 0.63 bpm, respectively) from 0 days (39.5 ± 0.65 bpm). Feed intake significantly decreased in heat stress groups (3 days and 6 days, 3.7 ± 0.14 kg and 4.0 ± 0.15 kg, respectively) than the control group (0 day, 5.0 ± 0.00 kg). In addition, final BW significantly decreased in heat stress groups (3 days and 6 days, 211.8 ± 4.75 kg and 215.5 ± 3.50 kg, respectively) than the control group (0 day, 240.0 ± 25.00 kg). However, heat stress has no significant effect on blood parameter, serum T3 and cortisol. Nevertheless, heat stress increased HSPs mRNA expression in liver tissue, and serum concentration of HSPs. Despite Hanwoo cattle may have high adaptive ability to heat stress, our results suggested that heat stress directly effect on body temperature and respiration rate as well as serum and tissue HSPs. Therefore, we are recommended that HSPs could be the most appropriate indicators of Hanwoo cattle response to heat stress.