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Saccharomyces cerevisiae Hsp30 is Necessary for Homeostasis of a Set of Thermal Stress Response Functions

  • Thakur, Suresh;Chakrabarti, Amitabha
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.403-409
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    • 2010
  • Saccharomyces cerevisiae Hsp30 is a plasma membrane heat shock protein that is induced by various environmental stress conditions. However, the functional role of Hsp30 during diverse environmental stressors is not presently known. To gain insight into its function during thermal stress, we have constructed and characterized a ${\Delta}hsp30$ strain during heat stress. $BY4741{\Delta}hsp30$ cells were found to be more sensitive compared with BY4741 cells, when exposed to a lethal heat stress at $50^{\circ}C$. When budding yeast is exposed to either heat shock or weak organic acid, it inhibits Pma1p activity. In this study, we measured the levels of Pma1p in mutant and Wt cells both during optimal temperature and heat shock temperature. We observed that $BY4741{\Delta}hsp30$ cells showed constitutive reduction of Pma1p. To gain further insights into the role of Hsp30 during heat stress, we compared the total protein profile by 2D gel electrophoresis followed by identification of differentially expressed spots by LC-MS. We observed that contrary to that expected from thermal-stress-induced changes in gene expression, the ${\Delta}hsp30$ mutant maintained elevated levels of Pdc1p, Trx1p, and Nbp35p and reduced levels of Atp2p and Sod1p during heat shock. In conclusion, Hsp30 is necessary during lethal heat stress, for the maintenance of Pma1p and a set of thermal stress response functions.

Suppression of Inflammation, Osteoclastogenesis and Bone Loss by PZRAS Extract

  • Li, Liang;Park, Young-Ran;Shrestha, Saroj Kumar;Cho, Hyoung-Kwon;Soh, Yunjo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1543-1551
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    • 2020
  • Panax ginseng has a wide range of activities including a neuroprotective effect, skin protective effects, enhanced DNA repairing, anti-diabetic activity, and protective effects against vascular inflammation. In the present study, we sought to discover the inhibitory effects of a mixture of natural products containing Panax ginseng, Ziziphus jujube, Rubi fructus, Artemisiae asiaticae and Scutellaria baicalensis (PZRAS) on osteoclastogenesis and bone remodeling, as neither the effects of a mixture containing Panax ginseng extract, nor its molecular mechanism on bone inflammation, have been clarified yet. PZRAS upregulated the levels of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GSH-R) and glutathione peroxidase (GSH-Px) and reduced malondialdehyde (MDA) in LPS-treated RAW264.7 cells. Moreover, treatment with PZRAS decreased the production of IL-1β and TNF-α. PZRAS also inhibited osteoclast differentiation through inhibiting osteoclastspecific genes like MMP-2, 9, cathepsin K, and TRAP in RANKL-treated RAW264.7 cells. Additionally, PZRAS has inhibitory functions on the RANKL-stimulated activation of ERK and JNK, which lead to a decrease in the expression of NFATc1 and c-Fos. In an in vivo study, bone resorption induced by LPS was recovered by treatment with PZRAS in bone volume per tissue volume (BV/TV) compared to control. Furthermore, the ratio of eroded bone surface of femurs was significantly increased in LPS-treated mice compared to vehicle group, but this ratio was significantly reversed in PZRAS-treated mice. These results suggest that PZRAS could prevent or treat disorders with abnormal bone loss.

Isolation of Flavonoids from Carthami Flos and their Antioxidative Activity (홍화의 플라보노이드 성분 분리 및 항산화 활성)

  • Chung, Sung-Hee;Moon, Ye-Ji;Kim, Sung-Gun;Kim, Kyoung-Young;Lee, Kyoung-Tae;Kim, Ho-Kyoung;Whang, Wan-Kyunn
    • YAKHAK HOEJI
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    • v.52 no.4
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    • pp.241-251
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    • 2008
  • In this study, isolation of antioxidative compounds was performed for development of anti-oxidizing agent. $CHCl_{3}$, $H_{2}O$, 30%, 60% MeOH, MeOH fractions were examined antioxidative activity by DPPH method, TBARS assay, and SOD like activity. It was revealed that 30%, 60% MeOH fractions had significant antioxidative activity. From 30%, 60% MeOH fraction, nine compounds were isolated and elucidated kaempferol $3-O-{\alpha}-L-rhamnopyranosyl$ $(1{\rightarrow}6)-{\beta}-D-glucopyranoside$ (1), quercetin $7-O-{\beta}-D-glucopyranoside$ (II), quercetin $3-O-{\alpha}-L-rhamnopyranosyl$ $(1{\rightarrow}6)$ ${\beta}-D-glucopyranoside(rutin)$ (III), 6-hydroxykaempferol $3-O-{\beta}-D-glucopyranoside$ (lV), kaempferol $3-O-{\beta}-D-glucopyranosyl$ $(1{\rightarrow}2)$ ${\beta}-D-glucopyranoside$ (V), kaempferol $3-O-{\beta}-D-glucopyranoside$ (VI), luteolin (VII), quercetin $3-O-{\beta}-D-glucopyranoside$ (VIII), apigenin $7-O-{\beta}-D-glucuronopyranoside$ (IX) through physicochemical data and spectroscopic methods (Negative FAB-MS, $^1H-NMR$, $^{13}C-NMR$). Entirely, all compounds had similar antioxidative activity, but more OH group had more antioxidative activity.

A 43 kD Protein Isolated from the Herb Cajanus indicus L Attenuates Sodium Fluoride-induced Hepatic and Renal Disorders in Vivo

  • Manna, Prasenjit;Sinha, Mahua;Sil, Parames C.
    • BMB Reports
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    • v.40 no.3
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    • pp.382-395
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    • 2007
  • The herb, Cajanus indicus L, is well known for its hepatoprotective action. A 43 kD protein has been isolated, purified and partially sequenced from the leaves of this herb. A number of in vivo and in vitro studies carried out in our laboratory suggest that this protein might be a major component responsible for the hepatoprotective action of the herb. Our successive studies have been designed to evaluate the potential efficacy of this protein in protecting the hepatic as well as renal tissues from the sodium fluoride (NaF) induced oxidative stress. The experimental groups of mice were exposed to NaF at a dose of 600 ppm through drinking water for one week. This exposure significantly altered the activities of the antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione reductase (GR) and the cellular metabolites such as reduced glutathione (GSH), oxidized glutathione (GSSG), total thiols, lipid peroxidation end products in liver and kidney compared to the normal mice. Intraperitoneal administration of the protein at a dose of 2 mg/kg body weight for seven days followed by NaF treatment (600 ppm for next seven days) normalized the activities of the hepato-renal antioxidant enzymes, the level of cellular metabolites and lipid peroxidation end products. Post treatment with the protein for four days showed that it could help recovering the damages after NaF administration. Time-course study suggests that the protein could stimulate the recovery of both the organs faster than natural process. Effects of a known antioxidant, vitamin E, and a non-relevant protein, bovine serum albumin (BSA) have been included in the study to validate the experimental data. Combining all, result suggests that NaF could induce severe oxidative stress both in the liver and kidney tissues in mice and the protein possessed the ability to attenuate that hepato-renal toxic effect of NaF probably via its antioxidant activity.

Anti-amnesic and Antioxidant Effect of Yeongkyekamjotanggayonggolmoryo Aqueous Extracts on Scopolamine-induced Memory Impairment in Mice (Scopolamine 유발 기억력손상 흰 쥐에서 영계감조탕가용골모려(笭桂甘棗湯加龍骨牡蠣)의 인지기능개선과 항산화 효과)

  • Kim, Dae-eok;Han, Da-young;Kim, Sang-ho;Chung, Dae-kyoo
    • Journal of Oriental Neuropsychiatry
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    • v.29 no.3
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    • pp.121-134
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    • 2018
  • Objectives: The purpose of this in vivo study is to observe anti-amnesic effects of Yeongkyekamjotanggayonggolmoryo (YGYM), a novel mixed herbal prescription, Ossis Mastodi and Ostreae Testa added Yeongkyekamjo-tang, on scopolamine induced amnesia in C57BL/6 mice through acetylcholine (ACh) and acetylcholinesterase (AChE) activity, Choline acetyltransferase (ChAT) mRNA expression, and antioxidant effects. Methods: Six groups, total 20 intact or 100 Sco treated mice were used in this study after one week of acclimatization period. Half the animals were used for passive avoidance task tests and hippocampus ACh content, AChE activity, and ChAT mRNA expression were measured. The other half was subjected to an underwater maze test and then the cerebral cortex antioxidant defense system was measured. Results: In the passive avoidance experiment, there was significant decrease in residence time in the bright room and in the underwater maze test, escape latency to escape from the esophagus significantly increased compared with the normal control group. At the final sacrifice, ACh content and ChAT mRNA expression decreased, AChE activity increased, and cerebral cortical MDA increased GSH content, SOD and CAT activity in Sco control mice, as compared to intact vehicle control mice. However, these Sco treatment-related memory loss through AChE activation destroyed the cerebral cortex antioxidant defense system, and was inhibited dose-dependently by 28 days consecutive oral pretreatments of YGYM extracts 500, 250, 125 mg/kg. Conclusions: In the above results, YGYM extract that oral administration of YGYM extracts alleviates the antioxidant defense system, through preservation of ACh mediated by upregulation of ChAT mRNA expression, and increase of AChE inhibition and brain antioxidant defense systems.

Comparison of Biological Activities on Rehmanniae Radix and Fermented Rehmanniae Radix (지황(地黃)과 발효(醱酵) 지황(地黃)의 생리활성 비교 연구)

  • Kim, Eun-Hyu;Kim, Kyoung-Shin;Chae, Suhn-Kee;Kim, Byoung-Soo;Kang, Jung-Soo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.3
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    • pp.306-313
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    • 2012
  • Herbal medicines are medicinal products containing a single or a mixture of two or more different herbal substances or herbal preparations as active principles. Recently, much attention has been paid to developing various kinds of fermented herbal extracts, a new type of traditional herbal medicine in the field of Korean traditional medicine. The fermentation of medicinal herbs is intended to exert a favorable influence on bioestability, bioavaliablilty and pharmacological activity of herbal extract in the gastrointestinal tract as well as intensifying the nutritional and pharmacological aspects of the medicinal herbs. The purpose of this study was to investigate biological activities of fermented Rehmanniae Radix by lactic acid bacteria at $30^{\circ}C$ for 3 days in comparison with those for Rehmanniae Radix The fermented Rehmanniae Radix exhibited different chemical profile to Rehmanniae Radix generated with HPLC, indicating production of new ingredients during fermentation. Rehmanniae Radix served as good nutritional sources for the growth of lactic acid bacteria showing increased number of bacteria during fermentation. Toxic effect of the fermented Rehmanniae Radix to cells were not seen judged by the MTT assay. The fermented Rehmanniae Radix exhibited better antioxidant effect than non-fermented Rehmanniae Radix analyzed by a SOD-likely assay. Both hypoglycemic and hypotensive effects of the fermented Rehmanniae Radix were also detected and better than those for Rehmanniae Radix in showing dose-dependent inhibitory effects on alpha-glucosidase and ACE, respectively. In conclusion, fermented Rehmanniae Radix appears to have more biological activities than non-feremented Rehmanniae Radix showing not only antioxidant effect but also cardiovascular protection.

Lonchocarpine Increases Nrf2/ARE-Mediated Antioxidant Enzyme Expression by Modulating AMPK and MAPK Signaling in Brain Astrocytes

  • Jeong, Yeon-Hui;Park, Jin-Sun;Kim, Dong-Hyun;Kim, Hee-Sun
    • Biomolecules & Therapeutics
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    • v.24 no.6
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    • pp.581-588
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    • 2016
  • Lonchocarpine is a phenylpropanoid compound isolated from Abrus precatorius that has anti-bacterial, anti-inflammatory, antiproliferative, and antiepileptic activities. In the present study, we investigated the antioxidant effects of lonchocarpine in brain glial cells and analyzed its molecular mechanisms. We found that lonchocarpine suppressed reactive oxygen species (ROS) production and cell death in hydrogen peroxide-treated primary astrocytes. In addition, lonchocarpine increased the expression of anti-oxidant enzymes, such as heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO1), and manganese superoxide dismutase (MnSOD), which are all under the control of Nrf2/antioxidant response element (ARE) signaling. Further, mechanistic studies showed that lonchocarpine increases the nuclear translocation and DNA binding of Nrf2 to ARE as well as ARE-mediated transcriptional activities. Moreover, lonchocarpine increased the phosphorylation of AMP-activated protein kinase (AMPK) and three types of mitogen-activated protein kinases (MAPKs). By treating astrocytes with each signaling pathway-specific inhibitor, AMPK, c-jun N-terminal protein kinase (JNK), and p38 MAPK were identified to be involved in lonchocarpine-induced HO-1 expression and ARE-mediated transcriptional activities. Therefore, lonchocarpine may be a potential therapeutic agent for neurode-generative diseases that are associated with oxidative stress.

Antitumor Activity and Antioxidant Role of Ichnocarpus frutescens Against Ehrlich Ascites Carcinoma in Swiss Albino Mice

  • Dash, Deepak K;Nayak, Siva S;Samanta, Soma;Ghosh, Tirtha;Jha, Tarun;Maiti, Bhim C;Maity, Tapan K
    • Natural Product Sciences
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    • v.13 no.1
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    • pp.54-60
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    • 2007
  • The plant Ichnocarpus frutescens (Linn) R.Br. (Family-Apocynaceae) has been indicated for the treatment of various diseases, one amongst it is cancer. The purpose of this study was to investigate experimentally the possible antitumor activity and antioxidant role of Ichnocarpus frutescens in the mice transplanted with Ehrlich ascites carcinoma (EAC). The chloroform and methanol extract of whole plant of Ichnocarpus frutescens (CEIF and MEIF) were administered intraperitoneally at the dose of 150 mg/kg and 300 mg/kg, body weight per day for 7 days after 24 h of tumor inoculation in mice. Treatment with CEIF at the dose of 150 mg/kg and 300 mg/kg remarkably decreased the tumor volume, packed cell volume, viable cell count and increased the nonviable cell count of EAC tumor bearing mice when compared to e effect of MEIF at 150 mg/kg and 300 mg/kg. Further the EAC mice treated with CEIF and MEIF showed significant decrease in the level of lipid peroxidation and significant increase in the level of antioxidant enzymes such as glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT), however the decreasing and increasing capacity of CEIF was less in both doses as compared to MEIF. Based on these results, it can be concluded that the chloroform and methanol extact of Ichnocarpus frutescens exhibit significant antitumor and antioxidant activity in EAC bearing mice.

Survival Rates and Physiological Response of Pacific Abalone Haliotis discus hannai Held in Live Fish Containers (적정 수송 조건하의 활어용 컨테이너 내 참전복(Haliotis discus hannai)의 생존 및 생리 변화 구명)

  • Yang, Sung Jin;Jun, Je-Cheon;Kang, Hee Woong;Park, Noh Back;Min, Byung Hwa
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.3
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    • pp.238-247
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    • 2018
  • This study investigated survival rates and physiological responses in Pacific abalone (Haliotis discus hannai) to 18 days of containment in live fish containers ($8^{\circ}C$, 34 psu). The investigation was divided into three periods: before, during, and recovery after transportation. The overall survival rate was greater than 99%. Glucose, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) rose immediately on the first day of containment, but then gradually returned to normal levels. $NH_3$ continued to rise after the first day, but during the recovery period it decreased to a level not significantly different from that of the control group. $Na^+$ and osmolality did not show any abnormal changes. After recovery, superoxide dismutase (SOD) was not significantly different from control. Abalone in the experimental group had lower glutathione reductase (GR) than control. The hyalinocyte ratio fell immediately after confinement, but then gradually increased until it reached a normal level. The ratios of apoptotic and necrotic cells indicated no specific variations in hemocyte viability. Histological changes in the epidermal layer and muscle layer of the foot were not significantly different from those seen in the control group. The experimental data obtained in this study suggest that live fish containers may be used for transport of Pacific abalone without significantly impacting their physiology or survival rates.

Comparisons of Chicken Muscles between Layer and Broiler Breeds Using Proteomics

  • Jung, K. C.;Jung, W. Y.;Lee, Y. J.;Yu, S. L.;Choi, K. D.;Jang, B. G.;Jeon, J. T.;Lee, J. H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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    • pp.307-312
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    • 2007
  • The present study was carried out to investigate differentially expressed chicken muscle proteins using proteomics approach. More than 300 protein spots were investigated for the muscle samples in 2DE gels and the differentially expressed protein spots between pectoralis and peroneus longus muscles from Cornish and White Leghorn breeds were characterized by MALDI-TOF. In pectoralis muscles, PGAM1 protein was detected as differentially expressed between White Leghorn and Cornish breeds. On the other hand, 4 protein spots (SP22, nxf-2, SOD1, TNNI2) were differentially expressed between White Leghorn and Cornish breeds in peroneus longus muscles. These proteins assumed to be related with muscle development, growth, stress, and movements in chicken. In this experimental process, 2D reference map of the chicken muscle proteins was needed and 25 proteins, which were commonly expressed in both pectoralis and peroneus longus muscles in both breeds, were selected and characterized. Upon finishing the exact roles of the differentially expressed proteins, the identified 5 proteins will be used as valuable information for the fundamental mechanisms of muscle biology and underline genetics.