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Hepatoprotective Effect of Bacillus subtilis-fermented Silkworm (Bombyx mori L.) Extract on Non-alcoholic Fatty Liver in Rats (고초균 발효누에 추출물이 비알코올성 지방간 유발 흰쥐에 미치는 간 기능 개선 효과)

  • Kim, Tae-Hoon;Ahn, Hee-Young;Kim, Young-Wan;Sim, So-Yeon;Cho, Hyun-Dong;Kim, Man-Do;Lee, You-Jung;Cho, Young-Su
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1031-1039
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    • 2017
  • The aim of this study was to investigate the potential effects of extracts from silkworm Bombyx mori L. fermented with Bacillus subtilis KACC 91157 at levels of 5%(v/w) and 10%(v/w) in Sprague-Dawley rats intoxicated with 1%(w/w) orotic acid (OA) for 10 days. The rats were divided into a normal group (N), a control group (C: OA), and treatment groups (SP10: OA + 10% extracts from B. mori L.; BSP5: OA + 5% extracts from B. mori L. fermented with B. subtilis KACC 91157; BSP10: OA + 10% extracts from B. mori L. fermented with B. subtilis KACC 91157). Serum activities of aspartate aminotransferase (AST), alanine transferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) increased following OA feeding, but the rise was slightly reduced by administration of BSP10. The total lipid, free fatty acid, phospholipid, total cholesterol, and triglyceride contents in serum were significantly lower in the OA treatment groups than in the N group. However, the contents slightly increased following the administration of BSP10. Glutathione concentrations in liver and serum were reduced in the OA-induced fatty liver, but they increased following the administration of BSP10. Hepatocytes in the OA-induced fatty liver contained numerous large droplets. However, SP10, BSP5, and BSP10 feeding prevented OA-induced lipid droplet accumulation in hepatocytes. Accordingly, extracts from silkworm powder fermented with B. subtilis could be an ideal material as a dietary supplement in healthy functional foods to improve the effects of fatty liver.

An Empirical Study of the Recovery Experiment in Clinical Chemistry (임상화학검사실에서 회수율 실험의 실증적 연구)

  • Chang, Sang-Wu;Lee, Sang-Gon;Song, Eun-Young;Park, Yong-Won;Park, Byong-Ok
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.184-188
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    • 2006
  • The purpose of the recovery experiment in clinical chemistry is performed to estimate proportional systematic error. We must know all measurements have some error margin in measuring analytical performance. Proportional systematic error is the type of error whose magnitude increases as the concentration of analyte increases. This error is often caused by a substance in the sample matrix that reacts with the sought for analyte and therefore competes with the analytical reagent. Recovery experiments, therefore, are used rather selectively and do not have a high priority when another analytical method is available for comparison purposes. They may still be useful to help understand the nature of any bias revealed in the comparison of kit experiments. Recovery should be expressed as a percentage because the experimental objective is to estimate proportional systematic error, which is a percentage type of error. Good recovery is 100.0%. The difference between 100 and the observed recovery(in percent) is the proportional systematic error. We calculated the amount of analyte added by multiplying the concentration of the analyte added solution by the dilution factor(mL standard)/(mL standard + mL specimen) and took the difference between the sample with addition and the sample with dilution. When making judgments on method performance, the observed that the errors should be compared to the defined allowable error. The average recovery needs to be converted to proportional error(100%/Recovery) and then compared to an analytical quality requirement expressed in percent. The results of recovery experiments were total protein(101.4%), albumin(97.4%), total bilirubin(104%), alkaline phosphatase(89.1%), aspartate aminotransferase(102.8), alanine aminotransferase(103.2), gamma glutamyl transpeptidase(97.6%), creatine kinase(105.4%), lactate dehydrogenase(95.9%), creatinine(103.1%), blood urea nitrogen(102.9%), uric acid(106.4%), total cholesterol(108.5), triglycerides(89.6%), glucose(93%), amylase(109.8), calcium(102.8), inorganic phosphorus(106.3%). We then compared the observed error to the amount of error allowable for the test. There were no items beyond the CLIA criterion for acceptable performance.

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Effects of Systemic and Focal Hypoxia on the Activities of Rostral Ventrolateral Medullary Neurons in Cats

  • Yan, Hai-Dun;Kim, Charn;Kim, Ji-Mok;Lim, Won-Il;Kim, Sang-Jeong;Kim, Jun
    • The Korean Journal of Physiology
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    • v.30 no.1
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    • pp.105-116
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    • 1996
  • Rendering the brain ischemic would evoke the cerebral ischemic reflex which is characterized by an arterial pressor response, apnea and bradycardia. Since the rostral ventrolateral medulla (RVLM) is known to play a key role in the maintenance of normal cardiopulmonary activity, during the cerebral ischemic reflex some cardiac related cells should be excited and respiration related cells inhibited. In this context, the responses of RVLM neurons to systemie and focal hypoxia were analyzed in the present study. Twenty-five adult cats of either sex were anesthetized with ${\alpha}-chloralose$ and the single neuronal activities were identified from RVLM area. For the induction of focal hypoxia in the recording site, sodium cyanide was applied iontophoretically and for systemic hypoxia the animal was ventilated with nitrogen gas for a twenty-second period. Cellular activities were analyzed in terms of their discharge pattern and responses to the hypoxia by using post-stimulus time and single-pass time histograms. Of eighteen cardiac related cells recorded from the RVLM area, twelve cells were excited by iontophoresed sodium cyanide and of twenty-five respiration related cells, fourteen cells were excited by iontophoresed sodium cyanide. Remaining cells were either inhibited or unaffected. Eight of fifteen cells tested with iontophoresed sodium lactate were excited and remaining seven cells were inhibited. Systemic hypoxia induced by nitrogen gas inhalation elevated the arterial blood pressure, but excited, inhibited or unaffected the single neuronal activities. Some cells showed initial excitation followed by inhibition during the systemic hypoxia. Bilateral vagotomy resulted in a decrease of arterial pressor response to the systemic hypoxia, and a slight decrease in the rhythmicity related to cardiac and/or respiratory rhythms. The single neuronal responses to either systemic or focal hypoxia were not affected qualitatively by vagotomy. From the above results, it was concluded that the majority of the cardiac- and respiration- related neurons in the rostral ventrolateral medulla be excited by hypoxia, not through the mediation of peripheral chemoreceptors, and along with the remaining inhibited cells, all these cells be involved in the mediation of cerebral ischemic reflex.

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Effect of pH on Organic Acid Production by Anaerobiospirillum succiniciproducens (Anaerobiospirillum succiniciproducens에 의한 유기산 생성에 미치는 pH의 영향)

  • 강귀현;류화원장호남
    • KSBB Journal
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    • v.10 no.5
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    • pp.568-574
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    • 1995
  • To investigate the effect of pH on organic acid production by Anaerobiospirillum succiniciproducens, an anaerobic fermentation was carried out by maintaining the pH of the fermentation broth at 5.8, 6.0, 6.4, 6.8, and 7.2. At various pHs, the concentrations of cell were $1.0∼1.9g/\ell$ which were two to three times higher than those of the other worker's results, and the maximum was obtained at pH 5.8. Substrate consumption was increased by increasing the pH in the range of pH 6.0 to 6.8, while the sugar consumption rate at both pH 5.8 and 7.2 was very slow. The total amount of 2M $Na_2C0_3$ added for adjustment of pH change due to organic acid production was maximum at pH 6.8. Changes of conductivity of the fermentation broth was very simillar to those of 2M $Na_2C0_3$ added at various pHs. Therefore, it is suggested that determination of the amount of organic acid in a broth can be possible by measuring the conductivity. The maximum production yield of lactate based on glucose was 64% for pH 7.2 and 32% for pH 6.8, respectively.

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Pulmonary Toxicity Assessment of Aluminum Oxide Nanoparticles via Nasal Instillation Exposure (비강내 점적 노출을 통한 산화 알루미늄 나노입자의 폐독성 평가)

  • Kwon, Jung-Taek;Seo, Gyun-Baek;Lee, Mimi;Kim, Hyun-Mi;Shim, Ilseob;Jo, Eunhye;Kim, Pilje;Choi, Kyunghee
    • Journal of Environmental Health Sciences
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    • v.39 no.1
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    • pp.48-55
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    • 2013
  • Objective: The use of nanoparticle products is expected to present a potential harmful effect on consumers. Also, the lack of information regarding inhaled nanoparticles may pose a serious problem. In this study, we addressed this issue by studying pulmonary toxicity after nasal instillation of Al-NPs in SD rats. Methods: The animals were exposed to Al-NPs at 1 mg/kg body weight (low dose), 20 mg/kg body weight (medium dose) and 40 mg/kg body weight (high dose). To determine pulmonary toxicity, bronchoalveolar lavage (ts.AnBAL) fluid analysis and histopathological examination were conducted in rats. In addition, cell viability was investigated at 24 hours after the treatment with Al-NPs. Results: BAL fluid analysis showed that total cells (TC) count and total protein (TP) concentrations increased significantly in all treatment groups, approximately two to three times. Also, lactate dehydrogenase (LDH) and cytokines such as TNF-alpha and IL-6 dose-dependently increased following nasal instillation of Al-NPs. However, polymorphonuclear leukocytes (PMNs) levels showed no significant changes in a dose dependant manner in BAL fluid. In the cytotoxicity analysis, the treatment of Al-NPs significantly and dose-dependently induced cell viability loss (20 to 30%) and damage of cell membrane (5 to 10%) in rat normal lung epithelial cells (L2). Conclusions: Our results suggest that inhaled Al-NPs in the lungs may be removed quickly by alveolar macrophages with minimal inflammatory reaction, but Al-NPs have the potential to affect lung permeability. Therefore, extensive toxicity evaluations of Al-NPs are required prior to their practical application as consumer products.

Protective Effect of 3,5-Dicaffeoylquinic Acid Isolated from Ligularia fischeri against Oxidative Damage in HepG2 Cells (HepG2 세포에서 산화적 손상에 대한 곰취 유래 3,5-Dicaffeoylquinic Acid의 보호 효과)

  • Park, Sun-Young;Kim, Gur-Yoo;Jhoo, Jin-Woo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.11
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    • pp.1286-1292
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    • 2017
  • This study was conducted to investigate the hepatoprotective effects of 3,5-dicaffeoylquinic acid (3,5-DCQA) isolated from Ligularia fischeri against hydrogen peroxide-induced oxidative stress in HepG2 cells. Antioxidative effects of 3,5-DCQA were determined by measuring antioxidant enzyme [superoxide dismutase (SOD), catalase (CAT) glutathione peroxidase (GPx)] expression levels against hydrogen peroxide-induced oxidative stress using real-time PCR analysis. 3,5-DCQA treatment significantly increased gene expression levels of SOD, CAT, and GPx in a dose-dependent manner ($10{\sim}30{\mu}g/mL$) in HepG2 cells. Hepatoprotective effects were analyzed by measuring glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT) activities using a biochemistry analyzer in hydrogen peroxide-treated HepG2 cells. 3,5-DCQA treatment significantly reduced GOT, LDH, and GGT activities in a dose-dependent manner ($10{\sim}30{\mu}g/mL$) against increased liver function index enzyme activities induced by hydrogen peroxide oxidative stress in HepG2 cells. The results reveal that 3,5-DCQA compound isolated from Ligularia fischeri can be useful for the development of an effective hepatoprotective agent.

Neuronal Cell Protective Effects of Methanol Extract from Cheonggukjang Using in vitro System (In vitro system에서 청국장 메탄올 추출물의 신경세포 보호효과)

  • Jeong, Chang-Ho;Kwak, Ji-Hyun;Kim, Ji-Hye;Choi, Gwi-Nam;Jeong, Hee-Rok;Heo, Ho-Jin
    • Korean Journal of Food Science and Technology
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    • v.42 no.6
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    • pp.768-772
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    • 2010
  • In this study, the neuronal cell protective effects of methanol extract from cheonggukjang were evaluated. The proximate composition and total phenolics of the methanol extract were 40.95% crude protein, 22.49% crude fat, 15.99% nitrogen free extract, 7.91% moisture, 6.74% crude ash, 5.92% crude fiber, and 28.43 mg/g of total phenolics. Intracellular ROS accumulation resulting from $H_2O_2$ treatment of PC12 cells was significantly reduced when methanol extract was present in the media compared to PC12 cells treated with $H_2O_2$ only. In a cell viability assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium-bromide (MTT), the methanol extract showed protective effects against $H_2O_2$-induced neurotoxicity, and lactate dehydrogenase (LDH) release into the medium was also inhibited. Furthermore, the inhibitory effect of the methanol extract against acetylcholinesterase was dose-dependent.

PC12 Cell Protective Effects of Broccoli (Brassica oleracea var. italica) Leaf Fraction against H2O2-induced Oxidative Stress (H2O2로 유발된 산화적 스트레스에 대한 브로콜리(Brassica oleracea var. italica) 잎 분획물의 PC12 cell 보호 효과)

  • Park, Seon Kyeong;Jin, Dong Eun;Park, Chang Hyeon;Seung, Tae Wan;Choi, Sung-Gil;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.46 no.4
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    • pp.483-488
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    • 2014
  • To examine the physiological effects of broccoli (Brassica oleracea var. italica) leaf, the bioavailable compounds in broccoli leaf extract, and its in vitro neuroprotective effects against $H_2O_2$-induced oxidative stress were examined in this study. The chloroform fraction of broccoli leaf extract had the highest total phenolic content of all the fraction than others, and the highest 2,2"-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical-scavenging activity and malondialdehyde (MDA) inhibitory effect. Intracellular reactive oxygen species (ROS) accumulation resulting in $H_2O_2$-treated in PC12 cells was significantly lower when the chloroform fraction was present in the medium compared to that in PC12 cells treated with $H_2O_2$ alone. In a cell viability assay performed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), the chloroform fraction showed protective effects against $H_2O_2$-induced neurotoxicity and inhibited lactate dehydrogenase (LDH) release into the medium. High-performance liquid chromatography (HPLC) analysis showed that ferulic acid was the predominant phenolic compound in chloroform fraction of broccoli leaf.

Anti-inflammatory Effect of Artemisia Capillaris Thunberg in Lipopolysaccharide-exposed Rats (인진호(茵蔯蒿)가 LPS 염증유발 흰쥐의 전염증성 cytokine 생산 및 혈액성상에 미치는 영향)

  • Seo, Yong-Seok;Lee, Eun;Cha, Yun-Yeop
    • Journal of Korean Medicine Rehabilitation
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    • v.20 no.3
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    • pp.27-35
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    • 2010
  • Objectives : The present study investigated anti-inflammatory effect of Artemisia Capilaris Thunberg in lipopolysaccharide-exposed rats. Methods : We divided lipopolysaccharide-exposed Sprague-Dawley rats into 4 groups. They were normal group, feed with 100 mg/kg Artemisia Capillaris Thunberg group, feed with 200 mg/kg Artemisia Capillaris Thunberg group and feed with 300 mg/kg Artemisia capilaris Thunberg group. They were administered for 6 weeks. We measured counts of red blood cell(RBC), the values of hemoglobin(Hb) and packed cell volume(PCV), plasma total protein concentration, albumin concentration, the ratio of albumin/globulin, the activities of plasma glutamic oxaloacetic transaminase(GOT), glutamic pyruvic transaminase(GPT), lactate dehydrogenase(LDH), the counts of white blood cell(WBC), the ratio of neutrophils, lymphocytes, monocytes, basophils, eosinophils, the concentration of plasma interleukin-$1{\beta}$($IL-1{\beta}$), plasma interleukin-6(IL-6), plasma tumor necrosis factor-$\alpha$($TNF-{\alpha}$), plasma interleukin-10(IL-10), the concentration of liver $IL-1{\beta}$ and IL-6, $TNF-{\alpha}$, IL-10. Results : Counts of RBC and the values of Hb and PCV, plasma total protein concentration and albumin concentration, the activities of plasma GOT, GPT and LDH showed no significant difference in the treatment groups. and the ratio of albumin/globulin was increased in Artemisia Capillaris Thunberg groups. The counts of WBC showed lower values in Artemisia Capillaris Thunberg groups than those of control group, In the ratio of neutrophils Thunberg groups. The ration of monocytes, basophils and eosinophils were below 5%, and showed no characteristic trend. The concentration of plasma interleukin-$1{\beta}$($IL-1{\beta}$), plasma inerleukin-6(IL-6) and plasma tumor necrosis factor-$\alpha$($TNF-{\alpha}$) showed a lower values in the Artemisia Capillaris Thunberg groups than those of control group, and the concentration of plasma interleukin-10(IL-10) showed no significant difference in the treatment groups. The concentration of liver $IL-1{\beta}$ and IL-6 showed a lower values in the Artemisia Capillaris Thunberg groups than those of control group, however the concentration of liver $TNF-{\alpha}$ and IL-10 showed no significant difference in the treatment groups. Conclusions : The Artemisia Capillaris Thunberg groups gives positive results of anti-inflammatory response by lipopolysaccharide(LPS) derivation.

Protective effects of perilla oil and alpha linolenic acid on SH-SY5Y neuronal cell death induced by hydrogen peroxide

  • Lee, Ah Young;Choi, Ji Myung;Lee, Myoung Hee;Lee, Jaemin;Lee, Sanghyun;Cho, Eun Ju
    • Nutrition Research and Practice
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    • v.12 no.2
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    • pp.93-100
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    • 2018
  • BACKGROUND/OBJECTIVE: Oxidative stress plays a key role in neuronal cell damage, which is associated with neurodegenerative disease. The aim of present study was to investigate the neuroprotective effects of perilla oil (PO) and its active component, alpha-linolenic acid (ALA), against hydrogen peroxide $(H_2O_2)$-induced oxidative stress in SH-SY5Y neuronal cells. MATERIALS/METHODS: The SH-SY5Y human neuroblastoma cells exposed to $250{\mu}M$ $H_2O_2$ for 24 h were treated with different concentrations of PO (25, 125, 250 and $500{\mu}g/mL$) and its major fatty acid, ALA (1, 2.5, 5 and $25{\mu}g/mL$). We examined the effects of PO and ALA on $H_2O_2$-induced cell viability, lactate dehydrogenase (LDH) release, and nuclear condensation. Moreover, we determined whether PO and ALA regulated the apoptosis-related protein expressions, such as cleaved-poly ADP ribose polymerase (PARP), cleaved caspase-9 and -3, BCL-2 and BAX. RESULTS: Treatment of $H_2O_2$ resulted in decreased cell viability, increased LDH release, and increase in the nuclei condensation as indicated by Hoechst 33342 staining. However, PO and ALA treatment significantly attenuated the neuronal cell death, indicating that PO and ALA potently blocked the $H_2O_2$-induced neuronal apoptosis. Furthermore, cleaved-PARP, cleaved caspase-9 and -3 activations were significantly decreased in the presence of PO and ALA, and the $H_2O_2$-induced up-regulated BAX/BCL-2 ratio was blocked after treatment with PO and ALA. CONCLUSIONS: PO and its main fatty acid, ALA, exerted the protective activity from neuronal oxidative stress induced by $H_2O_2$. They regulated apoptotic pathway in neuronal cell death by alleviation of BAX/BCL-2 ratio, and down-regulation of cleaved-PARP and cleaved caspase-9 and -3. Although further studies are required to verify the protective mechanisms of PO and ALA from neuronal damage, PO and ALA are the promising agent against oxidative stress-induced apoptotic neuronal cell death.