• Title/Summary/Keyword: protein tissues

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Immunotoxicological Effects of Aripiprazole: In vivo and In vitro Studies

  • Baek, Kwang-Soo;Ahn, Shinbyoung;Lee, Jaehwi;Kim, Ji Hye;Kim, Han Gyung;Kim, Eunji;Kim, Jun Ho;Sung, Nak Yoon;Yang, Sungjae;Kim, Mi Seon;Hong, Sungyoul;Kim, Jong-Hoon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.4
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    • pp.365-372
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    • 2015
  • Aripiprazole (ARI) is a commonly prescribed medication used to treat schizophrenia and bipolar disorder. To date, there have been no studies regarding the molecular pathological and immunotoxicological profiling of aripiprazole. Thus, in the present study, we prepared two different formulas of aripiprazole [Free base crystal of aripiprazole (ARPGCB) and cocrystal of aripiprazole (GCB3004)], and explored their effects on the patterns of survival and apoptosis-regulatory proteins under acute toxicity and cytotoxicity test conditions. Furthermore, we also evaluated the modulatory activity of the different formulations on the immunological responses in macrophages primed by various stimulators such as lipopolysaccharide (LPS), pam3CSK, and poly(I:C) via toll-like receptor 4 (TLR4), TLR2, and TLR3 pathways, respectively. In liver, both ARPGCB and GCB3004 produced similar toxicity profiles. In particular, these two formulas exhibited similar phospho-protein profiling of p65/nuclear factor $(NF)-{\kappa}B$, c-Jun/activator protein (AP)-1, ERK, JNK, p38, caspase 3, and bcl-2 in brain. In contrast, the patterns of these phospho-proteins were variable in other tissues. Moreover, these two formulas did not exhibit any cytotoxicity in C6 glioma cells. Finally, the two formulations at available in vivo concentrations did not block nitric oxide (NO) production from activated macrophage-like RAW264.7 cells stimulated with LPS, pam3CSK, or poly(I:C), nor did they alter the morphological changes of the activated macrophages. Taken together, our present work, as a comparative study of two different formulas of aripiprazole, suggests that these two formulas can be used to achieve similar functional activation of brain proteins related to cell survival and apoptosis and immunotoxicological activities of macrophages.

Restriction of Ca2+ deficiency-like symptoms by co-expressing a Ca2+ transporter and a Ca2+-binding protein in tomato (토마토에서 칼슘수송체와 칼슘결합단백질 공동발현에 의한 칼슘결핍유사증상의 완화)

  • Han, Jeung-Sul;Kang, Ho-Ju;Kim, Chang-Kil
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.549-555
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    • 2010
  • Here we focused on tip-burn and blossom-end rot (BER) symptoms in the tomato plants expressing the constitutively active form of $Ca^{2+}/H^+$ antiporter (sCAX1) and/or a Ca-binding protein (calreticulin, CRT) genes during their whole growth period. Conclusively we demonstrated that CRT is able to suppress the tip-burn and BER symptoms that were induced by sCAX1. Under poor nutrition condition, tomato plants overexpressing sCAX1 showed severe necrotic collapses in both roots and shoot polar tissues, which are in accordance with $Ca^{2+}$ deficient symptoms frequently observed in an agricultural cultivation of tomato. Reciprocal grafting trials using sCAX1 and wild type plants revealed that the tip-burn symptom by sCAX1 overexpression is not caused by hindrance of $Ca^{2+}$ uptake from soil. We constructed CRT overexpressing transgenic tomatoes, and crossed them with sCAX1 transgenic plants to investigate the effects of CRT on the symptoms of sCAX1 transgenic plants. Co-expression of sCAX1 and CRT significantly suppressed the $Ca^{2+}$ deficient symptoms of sCAX1 transgenic plants. Those results suggest the model that $Ca^{2+}$ homeostasis disturbed by the overexpression of sCAX1 may be suppressed by the co-expression of CRT.

Comparisons Among the Fishes of Genus Liobagrus in Korea by Their Morphology and Electrophoretic Patterns of Proteins (形態 및 蛋白質 電氣泳動像에 依한 韓國産 퉁가리屬 魚類의 比較)

  • 손영목;최의열;안태인
    • The Korean Journal of Zoology
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    • v.27 no.1
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    • pp.25-34
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    • 1984
  • Two species of fishes of Genus Liobagrus that had been collected from three separate river systems in Korea were compared by their morphology and gel electrophoretic patterns of water soluble proteins. In morphology L. andersoni from Han River was distinctly different from L. mediadiposalis collected from both Gum River and Nagdong River. But L. andersoni collected from Gum River not only showed partial similarity for both groups of fishes above in their classifying characteristics but also had a unique ratio of body width/standard length. Comparable similarity and difference among the three groups were also noted in their protein patterns of SDS polyacrylamide gel electrophoresis of various tissues. The difference was clearer and more distinct in the gel run with muscle proteins. A couple of more distinctly different low molecular polypeptides were detected by two dimensional gel electrophoresis. Since the protein patterns shown in this study are not only agreeable with the morphological results but also provide detailed comparisons, fishes of Liobagrus from various water sources can be classified reliably by gel electrophoresis. On the bases of tghe findings above, L. andersoni like fishes collected from Gum River should no longer be classified as L. andersoni. Before naming them as an independent species, the possibility of natural hybrid between the two defined species, or of a varient of L. andersoni by geograpic isolation should be tested.

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Inflammatory Effect of Light-Emitting Diodes Curing Light Irradiation on Raw264.7 Macrophage

  • Jeong, Moon-Jin;Kil, Ki-Sung;Lee, Myoung-Hwa;Lee, Seung-Yeon;Lee, Hye-Jin;Lim, Do-Seon;Jeong, Soon-Jeong
    • Journal of dental hygiene science
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    • v.19 no.2
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    • pp.133-140
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    • 2019
  • Background: The light-emitting diode (LED) curing light used is presumed to be safe. However, the scientific basis for this is unclear, and the safety of LED curing light is still controversial. The purpose of this study was to investigate the effect of LED curing light irradiation according to the conditions applied for the polymerization of composite resins in dental clinic on the cell viability and inflammatory response in Raw264.7 macrophages and to confirm the stability of LED curing light. Methods: Cell viability and cell morphology of Raw264.7 macrophages treated with 100 ng/ml of lipopolysaccharide (LPS) or/and LED curing light with a wavelength of 440~490 nm for 20 seconds were confirmed by methylthiazolydiphenyl-tetrazolium bromide assay and microscopic observation. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was confirmed by NO assay and $PGE_2$ enzyme-linked immunosorbent assay kit. Expression of interleukin $(IL)-1{\beta}$ and tumor necrosis factor $(TNF)-{\alpha}$ in total RNA and protein was confirmed by reverse transcription polymerase chain reaction and Western blot analysis. Results: The LED curing light did not affect the viability and morphology of normal Raw264.7 cells but affected the cell viability and induced cytotoxicity in the inflammation-induced Raw264.7 cells by LPS. The irradiation of the LED curing light did not progress to the inflammatory state in the inflammation-induced Raw264.7 macrophage. However, LED curing light irradiation in normal Raw264.7 cells induced an increase in NO and $PGE_2$ production and mRNA and protein expression of $(IL)-1{\beta}$ and $(TNF)-{\alpha}$, indicating that it is possible to induce the inflammatory state. Conclusion: The irradiation of LED curing light in RAW264.7 macrophage may induce an excessive inflammatory reaction and damage oral tissues. Therefore, it is necessary to limit the long-term irradiation which is inappropriate when applying LED curing light in a dental clinic.

Effect of Fish Meal Replacement on Insulin-like Growth Factor-I Expression in the Liver and Muscle and Implications for the Growth of Olive Flounder Paralichthys olivaceus (사료의 어분함량대체가 넙치(Paralichthys olivaceus)의 간과 근육 내 인슐린유사성장인자의 발현과 체성장에 미치는 영향)

  • Park, Su-Jin;Moon, Ji-Sung;Seo, Jin-Song;Nam, Taek-Jeong;Lee, Kyeong-Jun;Lim, Sang-Gu;Kim, Kang-Woong;Lee, Bong-Joo;Hur, Sang-Woo;Choi, Youn Hee
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.2
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    • pp.141-148
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    • 2019
  • This study examined the effect of insulin-like growth factor (IGF)-I expression in the liver and muscle on the growth of Paralichthys olivaceus fed diets low in fish meal. A feeding experiment was conducted at Jeju National University, Jeju Island, Korea. Groups of P. olivaceus (total initial weight: 200 g) were maintained for 20 weeks on one of five experimental diets containing different proportions of fish meal. Diets containing 0%, 20%, 30%, 40%, and 50% fish meal were labeled FM0, FM20, FM30, FM40, and FM50, respectively. Fish growth was observed every 4 weeks during the feeding experiment, and plasma and liver and muscle tissues were sampled. Plasma IGF-I levels were analyzed using an ELISA kit. The mechanism of IGF-I receptor signaling was examined using immunoblotting and reverse transcription-polymerase chain reaction. The greatest total weight increase was observed in the FM30 group. In parallel, plasma levels of IGF-I and IGF-binding protein were highest in the FM30 group, and mRNA and protein expression were also significantly higher in this group. The first step in the IGF-I signaling pathway, tyrosine-phosphorylation checking, occurred smoothly until 20 weeks. These results suggest that a dietary ratio of 30% fish meal best promotes growth in this species. The IGF-I signaling pathway in the liver and muscle is associated with growth in P. olivaceus.

Lethal (2) Essential for Life [l(2)efl] Gene in the Two-spotted Cricket, Gryllus bimaculatus (Orthoptera: Gryllidae) (쌍별귀뚜라미(Gryllus bimaculatus)의 l(2)efl cDNA 클로닝과 발현분석)

  • Kwon, Kisang;Lee, Nuri;Kwon, O-Yu
    • Journal of Life Science
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    • v.31 no.7
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    • pp.671-676
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    • 2021
  • A cDNA encoding the protein lethal (2) essential for life [l(2)efl] was cloned from Gryllus bimaculatus and named GBl(2)efl. This protein is composed of 189 amino acids, including an N-glycosylation site and 15 phosphorylation sites. Its predicted molecular mass is 21.19 kDa, with a theoretical isoelectric point of 6.2. The secondary structure of GBl(2)efl was predicted from the identification of random coils (56.08%), alpha helices (22.22%), extended strands (17.99%), and beta turns (3.7%) through sequence analyses. A homology analysis revealed that GBl(2)efl exhibited a high similarity with other species at the amino acid level, ranging from 52% to 69%. While GBl(2)efl mRNA expression was higher in the dorsal longitudinal flight muscle following a three-day starvation and in the Malpighian tubules following a one-day starvation, no changes in expression were detected in other tissues. Furthermore, tunicamycin-induced endoplasmic reticulum (ER) stress resulted in an approximately 1.8-fold higher expression in the fat body compared with the wild type.

Effect of serotonin on the cell viability of the bovine mammary alveolar cell-T (MAC-T) cell line

  • Xusheng, Dong;Chen, Liu;Jialin, Miao;Xueyan, Lin;Yun, Wang;Zhonghua, Wang;Qiuling, Hou
    • Journal of Animal Science and Technology
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    • v.64 no.5
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    • pp.922-936
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    • 2022
  • 5-Hydroxytryptamine (5-HT), a monoamine, as a local regulator in the mammary gland is a chemical signal produced by the mammary epithelium cell. In cows, studies have shown that 5-HT is associated with epithelial cell apoptosis during the degenerative phase of the mammary gland. However, studies in other tissues have shown that 5-HT can effectively promote cell viability. Whether 5-HT could have an effect on mammary cell viability in dairy cows is still unknown. The purpose of this study was to determine: (1) effect of 5-HT on the viability of bovine mammary epithelial cells and its related signaling pathways, (2) interaction between prolactin (PRL) and 5-HT on the cell viability. The bovine mammary alveolar cell-T (MAC-T) were cultured with different concentrations of 5-HT for 12, 24, 48 or 72 hours, and then were assayed using cell counting kit-8, polymerase chain reaction (PCR) and immunobloting. The results suggested that 20 μM 5-HT treatment for 12 or 24 h promote cell viability, which was mainly induced by the activation of 5-HT receptor (5-HTR) 1B and 4, because the increase caused by 5-HT vanished when 5-HTR 1B and 4 was blocked by SB224289 and SB204070. And protein expression of mammalian target of rapamycin (mTOR), eukaryotic translation elongation factor 2 (eEF2), janus kinase 2 (JAK2) and signal transducer and activator of transcription 5 (STAT5) were decreased after blocking 5-HT 1B and 4 receptors. When MAC-T cells were treated with 5-HT and PRL simultaneously for 24 h, both the cell viability and the level of mTOR protein were significantly higher than that cultured with 5-HT or PRL alone. In conclusion, our study suggested that 5-HT promotes the viability of MAC-T cells by 5-HTR 1B and/or 4. Furthermore, there is a reciprocal relationship between PRL and 5-HT.

Studies on Antioxidant and Anti-hyperlipidemic Effects in Induced Hypercholesterolemic Rats of Steamed Achyranthis Bidentatae Radix (우슬(牛膝)의 증숙(蒸熟)에 따른 항산화 효능 및 고지혈증 유발 흰쥐에 대한 고지혈증 개선 효능에 대한 연구)

  • Kim, Jeong-Soo;Park, MyungJae;Kim, SuJi;Shin, Mi-Rae;Lee, AhReum;Park, Hae-Jin;Roh, Seong-Soo;Seo, Young Bae
    • The Korea Journal of Herbology
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    • v.33 no.6
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    • pp.43-60
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    • 2018
  • Objectives : This study was conducted to compare the effect of Achyranthis Bidentatae Radix on antioxidant changes and antihyperlipidemics in hypercholesterolemic rats according to changes in the number of steams. Methods : The experimental animals were divided into six groups: normal (Nor), high cholesterol diet (Veh), high cholesterol diet with Simvastatin 5 mg/kg (Sim), high cholesterol diet with AR0 extract 200 mg/kg (AR0), high cholesterol diet with AR6 extract 200 mg/kg (AR6) and high cholesterol diet plus AR9 extract 200 mg/kg (AR9). I measured HDL, LDL TC, TG contents and ROS from each serums and compared them. I analyzed antioxidant-related protein expressions and cholesterol-related protein expressions in each liver tissues. I also performed optical microscopic analysis through liver tissue staining. Results : The Antioxidant activity of Achyranthis Bidentatae Radix was increased by steaming processes. In vivo, AR0 decreased liver weight in hyperlipidemic rats. LDL TC, TG, AI and CRF were decreased but HDL was increased in AR0. AST, ALT, creatinine, and BUN were decreased in AR0. ROS was decreased in AR0, AR6, and AR9. SOD, catalase and GPx were decreased by steaming processes. p-AMPK was decreased but SREBP-2, p-ACC, and HMGCR were increased by steaming processes. Histologic analysis showed that AR0 decreased liver fat accumulation. Conclusion : In conclusion, AR0 can be developed as a treatment for hyperlipemia. And it is required to research on antioxidant effect changes among different times steamed Achyranthis Bidentatae Radix.

Effect of Ginsenoside Rg3 on COX-2 Expression in Brain Tissue of Lipopolysaccharide-Treated Mice (Ginsenoside Rg3이 Lipopolysaccharide에 의한 생쥐 뇌조직의 Cyclooxygenase-2 발현에 미치는 영향)

  • Choi, Wonik;Cho, Yong-Deok;Lee, Joon-Seok;Shin, Jung-Won;Kim, Seong-Joon;Sohn, Nak-Won
    • The Korea Journal of Herbology
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    • v.27 no.6
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    • pp.131-137
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    • 2012
  • Objectives : Cyclooxygenase (COX) plays a central role in the inflammatory cascade by converting arachidonic acid into prostaglandin. COX-2 is typically induced by inflammatory stimuli in the majority of tissues, it is responsible for propagating the inflammatory response and thus, considered as the best target for anti-inflammatory drugs. The present study investigated the modulatory effect of ginsenoside Rg3, a principle active ingredient in Panax ginseng, on COX-2 expression in the brain tissue induced by systemic lipopolysaccharide (LPS) treatment in C57BL/6 mice. Methods : Because systemic LPS treatment induces COX-2 expression immediately in the brain, ginsenoside Rg3 was treated orally with doses of 10, 20, and 30 mg/kg at 1 hour before the LPS (3 mg/kg, i.p.) injection. At 4 hours after the LPS injection, COX-2 mRNA was measured by real-time polymerase chain reaction method, COX-2 protein levels were measured by Western blotting. In addition, COX-2 expressions in brain tissue were observed with immunohistochemistry and double immunofluoresence labeling. Results : Ginsenoside Rg3 (20 and 30 mg/kg) significantly attenuates up-regulation of COX-2 mRNA and protein expression in brain tissue at 4 hours after the LPS injection. Moreover, ginsenoside Rg3 (20 mg/kg) significantly reduced the number of COX-2 positive neurons in the cerebral cortex and amygdala. Conclusion : These results indicate that ginsenoside Rg3 plays a modulatory role in neuroinflammation through the inhibition of COX-2 expression in the brain and suggest that ginsenoside Rg3 and ginseng may be effective on neurodegenerative diseases caused by neuroinflammation.

Antioxidant and Anti-Obesity Effect of SM17 in High-Fat Diet Induced C57BL/6 Mice (고지방식이로 비만을 유도한 C57BL/6 마우스에서 SM17의 항산화 및 항비만 효과)

  • Kim, Soo Hyun;Kim, Su Ji;Kim, Kyeong Jo;Lee, Ah Reum;Roh, Seong-Soo;Lee, Young Cheol
    • The Korea Journal of Herbology
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    • v.32 no.5
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    • pp.47-55
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    • 2017
  • Objectives : Obesity is caused by the excess accumulation of fat in the body due to energy imbalance, and it causes various diseases. The aim of this study was to investigate an anti-obesity efficacy and an antioxidant activity of water from herbal mixture extract (SM17). Methods : The antioxidant activities were evaluated through radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radicals. To evaluated anti-obesity effect of SM17, we used a high fat diet fed mouse model. The SM17 (150 mg/kg body weight/day, p.o.) was treated every day for 6 weeks to C57BL/6 mice. Body weight and food intake were measured every day. The changes of reactive oxygen species (ROS), alanine aminotransferanse (ALT), aspartate aminotransferase (AST), triglycerids (TG) and total cholesterol (TC) in serum were analyzed after experiment. Also, expression of lipid metabolism related proteins were investigated by western blot analysis. Results : It was effective in antioxidant measurements, SM17 administration inhibited the biomarkers of lipid metaboism in serum and tissues. The administration of SM17 showed a significant reduction of body and tissue weight. Morever, it decreased ROS, ALT, AST, TG and TC in serum, compared with those of the obese mice. Adipogenesis-related protein expressions increased in obese mice compared to normal mice. However, SM17 group exhibited the down-expression of these proteins. Conclusion : A SM17 aqueous extract has a great effect on the stimulation (AMPK) activation, and may have a benefit to reduce a fatty acid metabolism through inhibition of lipid accumulation.