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Purification of Alginate Lyase from Streptomyces violaceoruber and the Growth Activity of Intestinal Bacteria by Degree of Polymerization of Alginate Hydrolysates (Streptomyces violaceoruber 유래 Alginate Lyase의 정제 및 Sodium Alginate 가수분해 올리고당의 중합도별 Bifidobacterium spp.과 Lactobacillus spp.에 대한 생육활성)

  • Yoon, Min;Park, Young-Seo;Park, Gwi-Gun
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.103-109
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    • 2017
  • Alginate lyase from Streptomyces violaceoruber was purified by DEAE sephacel chromatography and SP sepharose chromatography. The specific activity of the purified enzyme was 14.6 units/mg protein, representing a 40.6-fold purification of the crude extract. The final preparation thus obtained showed a single band on Tricine-SDS polyacrylamide gel electrophoresis whose molecular weight was determined to be 23.3 kDa. The polyMG block of sodium alginate was hydrolyzed by the purified alginate lyase and then separated by activated carbon column chromatography and bio gel P-2 gel filtration. The main hydrolysates were composed of hetero type M/G-oligosaccharides with the degrees of polymerization (D.P.) being 6 and 8. To investigate the effects of hetero type M/G-oligosaccharides from the sodium alginate on the growth of some intestinal bacteria, cells were cultivated individually on the modified-MRS medium containing D.P. 6 and 8 M/G-oligosaccharides. B. longumgrew 4.25-fold and 6.44-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides compared with those of standard MRS medium. In addition, B. bifidumgrew 3.3-fold and 5.4-fold more effectively by the treatment of D.P. 6 and 8 M/G-oligosaccharides. In conclusion, D.P. 8 was more effective than D.P. 6 hetero M/G-oligosaccharides as regards the growth of Bifidobacteriumspp. and Lactobacillus spp.

Effects of Dietary Bee Venom on Serum Characteristic, Antioxidant Activity and Liver Fatty Acid Composition in Broiler Chickens (정제 봉독의 급여가 육계의 혈청성상, 항산화능 그리고 간 내 지방산 조성에 미치는 효과)

  • Kim, Da-hye;Han, Sang-mi;Choi, Yun-Sang;Kang, Hwan-Ku;Lee, Hong-Gu;Lee, Kyung-woo
    • Korean Journal of Poultry Science
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    • v.46 no.1
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    • pp.39-46
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    • 2019
  • This study was conducted to investigate the effect of dietary bee venom on serum characteristics, antioxidant activity, and hepatic fatty acid composition in broiler chickens. A group of 875 one-day-old feather-sexed male broiler chicks were randomly allocated to five treatments with seven replicates (25 birds/replicate) for three weeks. A corn-soybean meal-based diet was used as the basal diet. Five dietary treatments were compared: 1) basal diet, 2) basal diet containing $10{\mu}g/kg$ of bee venom powder, 3) basal diet containing $50{\mu}g/kg$ of bee venom powder, 4) basal diet containing $100{\mu}g/kg$ of bee venom powder, and 5) basal diet containing $500{\mu}g/kg$ of bee venom powder. At 21 days, one bird per pen was slaughtered by asphyxiation in $CO_2$ gas, and blood was collected to measure serum characteristics and antioxidant activity. In addition, the liver was excised to measure the concentration of malondialdehyde and determine fatty acid composition. Increasing dietary bee venom in the diet failed to affect most serum parameters except for triglyceride and non-esterified fatty acids. Dietary bee venom inclusion quadratically increased the concentration of stearic acid (P<0.05), but decreased palmitoleic acid, oleic acid, linoleic acid, mono-unsaturated fatty acids, and poly-unsaturated fatty acids. Finally, dietary bee venom tended to lower hepatic malondialdehyde contents quadratically (P=0.054). In conclusion, our study revealed that dietary bee venom improved antioxidant capacity and affected fatty acid metabolism in broiler chickens.

Anti-proliferative Properties of p-Coumaric Acid in SNU-16 Gastric Cancer Cells (SNU-16 위암 세포주에서 p-coumaric acid의 세포성장 억제 효과)

  • Jang, Mi Gyeong;Ko, Hee Chul;Kim, Se-Jae
    • Journal of Life Science
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    • v.29 no.7
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    • pp.809-816
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    • 2019
  • The ubiquitous plant metabolite p-coumaric acid (p-CA) has antioxidant and anti-inflammatory properties, but its anti-cancer activity has not been established in gastric cancer cell lines. In this study, we investigated the effects of p-CA on the proliferation and transcriptome profile of SNU16 gastric cancer cells. Treatment with p-CA induced apoptosis of the SNU-16 cells by regulating the expression of pro-apoptotic and anti-apoptotic proteins, such as Bcl-2, poly (ADP-ribose) polymerase (PARP), Bax, procaspase-3, and cleaved-caspase-3. The genes differentially expressed in response to p-CA treatment of the SNU-16 cells were identified by RNA sequencing analysis. Genes regulated by p-CA were involved mainly in the inflammatory response, apoptotic processes, cell cycle, and immune response. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that the phosphatidylinositol-3-kinase-Akt and cancer signaling pathways were altered by p-CA. Protein-protein interaction (PPI) network analysis also revealed that p-CA treatment was correlated with differential expression of genes associated with the inflammatory response and cancer. Collectively, these results suggest that p-CA has potential utility in gastric cancer prevention.

Antioxidant and Anti-inflammatory Effects of Ethanol Extract of Aster yomena in RAW 264.7 Macrophages (RAW 264.7 대식세포에서 쑥부쟁이 추출물의 항산화 및 항염증 효능에 관한 연구)

  • Kim, Sung Ok;Jeong, Ji-Suk;Choi, Yung Hyun
    • Journal of Life Science
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    • v.29 no.9
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    • pp.977-985
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    • 2019
  • Aster yomena (Kitam.) Honda is an edible vegetable and perennial herb belonging to the Asteraceae family, and has been used for a long time for the prevention and treatment of various diseases. Although leaf extracts of A. yomena are known to have antioxidant and anti-inflammatory effects, accurate efficacy assessments are still inadequate. In this study, we investigated whether the antioxidant efficacy of ethanol extract of A. yomena leaf (EEAY) is correlated with the anti-inflammatory effect in RAW 264.7 macrophages. The results showed that EEAY significantly inhibited the hydrogen peroxide ($H_2O_2$)-induced growth inhibition in RAW 264.7 cells, which was associated with increased expression of nuclear factor erythroid 2-related factor-2 (Nrf2) and heme oxygenase-1 (HO-1). EEAY pretreatment also effectively prevented $H_2O_2$-induced reactive oxygen species generation and apoptosis through inhibition of caspase-3 activation and poly (ADP-ribose) polymerase degradation. Additionally, EEAY significantly increased the expression and production of interleukin-10, a representative anti-inflammatory cytokine, which was associated with increased expression of toll-like receptor 4 and myeloid differentiation factor 88 at transcriptional and translational levels. Furthermore, the increased production of nitric oxide (NO) by lipopolysaccharide was markedly abolished under the condition of EEAY pretreatment, and the inhibitory effect of NO production by EEAY was further increased by hemin, an HO-1 inducer. Overall, our results suggest that EEAY is able to activate the Nrf2/HO-1 signaling pathway to protect RAW 264.7 macrophages from oxidative and inflammatory stress.

Ziyuglycoside II Attenuates Tumorigenesis in Experimental Colitis-associated Colon Cancer (AOM/DSS로 유도된 마우스 대장암 모델에서의 Ziyuglycoside-II의 항염증효과)

  • Cheon, Hye-Jin;Kim, Jin-Kyung
    • Journal of Life Science
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    • v.29 no.9
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    • pp.941-948
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    • 2019
  • Colorectal cancer is a major health problem in industrialized countries. Ziyuglycoside II ($3{\beta}-3-{\alpha}$-1- arabinopyranosyloxy-19-hydroxyurs-12-en-28-oicacid), a triterpenoid saponin isolated from the roots of Sanguisorba officinalis L., possesses antioxidant, antiangiogenic, and anticancer properties. However, the therapeutic function of ziyuglycoside II in colitis-associated colorectal carcinogenesis is undefined. In the present study, the effect of ziyuglycoside II on colitis-associated colon cancer induced in mice using azoxymethane (AOM)/dextran sulfate sodium (DSS) was explored. The AOM model recapitulates many features of human colon cancer, but it lacks an inflammatory component. DSS induces colitis and promotes AOM-induced colon cancer in mice. BALB/c mice were injected with AOM and administered 2% DSS in drinking water. The mice were given ziyuglycoside II (1 or 5 mg/kg) orally three times per week, and colonic tissue was collected at 64 days. Administration of ziyuglycoside II markedly diminished the formation of colonic tumors. Western blot and immunohistological analyses showed that ziyuglycoside II noticeably decreased nuclear factor kappa-B-positive cells and levels of inflammation-related proteins, such as inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-${\alpha}$, and interleukin-6 in colon tissue. It also prompted apoptosis. Ziyuglycoside II treatment augmented cleaved forms of caspase-3, caspase-7, and poly (ADP-ribose) polymerase in colonic tissues. In conclusion, ziyuglycoside II could defend against colitis-associated tumorigenesis in mice by inhibiting inflammation and inducing apoptosis. This shows a promising chemopreventive potential for its use in colitis-associated colon cancer.

Influence of various serum supplement on in vitro culture for goat embryos (다양한 혈청 물질의 첨가가 염소 수정란의 체외배양에 미치는 영향)

  • Kim, Kwan-Woo;Jeon, Dayeon;Lee, Jinwook;Lee, Sung-Soo;Kim, Seungchang;Kim, Chan-Lan;Lee, Sang-Hoon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.9
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    • pp.510-516
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    • 2019
  • This study examined the effects of fetal bovine serum (FBS), goat blood serum (gBS), and poly-vinyl alcohol (PVA) on the in vitro development and embryo quality of goats for an improvement of embryo production. For the experiment, an in vitro fertilized embryo culture medium was supplemented with 10% FBS, 10% gBS, and 10% PVA to determine their effects on the embryo development efficiency and blastocyst quality. The results showed that the non-serum supplementation group showed significantly lower cleavage rate and blastocyst formation. On the other hand, the gBS and PVA supplementation groups showed a significant increase in the cleavage rate and better blastocyst formation than the control and FBS supplementation group. Furthermore, a TUNEL assay performed to confirm the blastocyst quality showed the same pattern as the embryo development experiment. These results showed that the supplemented gBS or PVA was more efficient in enhancing the in vitro development efficiency of goats than the supplementation of FBS or non-serum. On the other hand, considering the risk of an unidentified factor in gBS, PVA appears to be safer and more efficient in the in vitro development of goat embryos.

Molecular Characterization and Expression Analysis of Nucleoporin 210 (Nup210) in Chicken

  • Ndimukaga, Marc;Bigirwa, Godfrey;Lee, Seokhyun;Lee, Raham;Oh, Jae-Don
    • Korean Journal of Poultry Science
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    • v.46 no.3
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    • pp.185-191
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    • 2019
  • Nucleoporin 210 (Nup210) is associated with several physiological processes including muscle and neural cell differentiation, autoimmune diseases, and peripheral T cell homeostasis. Chicken Nup210 (chNup210) gene was originally identified as one of the differentially expressed genes (DEGs) in the kidney tissues of chicken. To elucidate the role of Nup210 in metabolic disease of chicken, we studied the molecular characteristics of chNup210 and analyzed its gene expression under the stimulation of Toll-like receptor 3 (TLR3) ligands. The Nup210 genomic DNA and amino acid sequences of various species including fowls, fishes, and mammals were retrieved from the Ensemble database and subjected to bioinformatics analyses. The expression of Nup210 from several chicken tissues was probed through qRT-PCR, and chicken fibroblast DF-1 cell line was used to determine the change in expression of chNup210 after stimulation with TLR3 ligand, polyinosinic-polycytidylic acid (poly (I:C)). The chNup210 gene was highly expressed in chicken lung and spleen tissues. Although highly conserved among the species, chNup210 was evolutionary clustered in the same clade as that of duck compared to other mammals. Furthermore, this study revealed that chNup210 is expressed in TLR3 signaling pathway and provides fundamental information on Nup210 expression in chicken. Future studies that offer insight into the involvement of chNup210 in the chicken innate immune response against viral infection are recommended.

Protective Effects of Quercetin-3-O-glucuronide against 1-methyl-4-phenylpyridinium-induced Neurotoxicity (1-methyl-4-phenylpyridinium으로 유도된 신경 손상에 대한 quercetin-3-O-glucuronide의 보호 효과)

  • Pariyar, Ramesh;Bastola, Tonking;Seo, Jungwon
    • Journal of Life Science
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    • v.29 no.2
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    • pp.191-197
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    • 2019
  • Parkinson's disease (PD) is a progressive neurodegenerative disease that mainly affects motor system with clinical features such as bradykinesia, rigidity, tremor and abnormal posture. PD is characterized by the death of dopaminergic neurons in the substantia nigra pars compacta, which is associated with accumulation of oxidative stress and dysregulation of intracellular signaling pathway. Quercetin-3-O-glucuronide (Q3GA), a major metabolite of quercetin, has been reported to have neuroprotective effects. In this study, we examined the neuroprotective effect of Q3GA against 1-methyl-4-phenyl pyridinium ($MPP^+$)-induced neurotoxicity of PD and the underlying molecular mechanisms in SH-SY5Y cells. MTT and LDH assay showed that Q3GA significantly decreased $MPP^+$-induced cell death, which is accompanied by a reduction in poly (ADP-ribose) polymerase (PARP) cleavage. Furthermore, it attenuated $MPP^+$-induced intracellular reactive oxygen species (ROS) with the reduction of Bax/ Bcl-2 ratio. Moreover, Q3GA significantly increased the phosphorylation of Akt and cAMP response element binding protein (CREB), but it has no effects on the phosphorylation of extracellular signal-regulated kinase (ERK). Taken together, these results demonstrate that Q3GA significantly attenuates $MPP^+$-induced neurotoxicity through ROS reduction and Akt/CREB signaling pathway in SH-SY5Y cells. Our findings suggest that Q3GA might be one of the potential candidates for the prevention and/or treatment of PD.

Cell-Based Screen Using Amyloid Mimic β23 Expression Identifies Peucedanocoumarin III as a Novel Inhibitor of α-Synuclein and Huntingtin Aggregates

  • Ham, Sangwoo;Kim, Hyojung;Hwang, Seojin;Kang, Hyunook;Yun, Seung Pil;Kim, Sangjune;Kim, Donghoon;Kwon, Hyun Sook;Lee, Yun-Song;Cho, MyoungLae;Shin, Heung-Mook;Choi, Heejung;Chung, Ka Young;Ko, Han Seok;Lee, Gum Hwa;Lee, Yunjong
    • Molecules and Cells
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    • v.42 no.6
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    • pp.480-494
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    • 2019
  • Aggregates of disease-causing proteins dysregulate cellular functions, thereby causing neuronal cell loss in diverse neurodegenerative diseases. Although many in vitro or in vivo studies of protein aggregate inhibitors have been performed, a therapeutic strategy to control aggregate toxicity has not been earnestly pursued, partly due to the limitations of available aggregate models. In this study, we established a tetracycline (Tet)-inducible nuclear aggregate (${\beta}23$) expression model to screen potential lead compounds inhibiting ${\beta}23$-induced toxicity. High-throughput screening identified several natural compounds as nuclear ${\beta}23$ inhibitors, including peucedanocoumarin III (PCIII). Interestingly, PCIII accelerates disaggregation and proteasomal clearance of both nuclear and cytosolic ${\beta}23$ aggregates and protects SH-SY5Y cells from toxicity induced by ${\beta}23$ expression. Of translational relevance, PCIII disassembled fibrils and enhanced clearance of cytosolic and nuclear protein aggregates in cellular models of huntingtin and ${\alpha}$-synuclein aggregation. Moreover, cellular toxicity was diminished with PCIII treatment for polyglutamine (PolyQ)-huntingtin expression and ${\alpha}$-synuclein expression in conjunction with 6-hydroxydopamine (6-OHDA) treatment. Importantly, PCIII not only inhibited ${\alpha}$-synuclein aggregation but also disaggregated preformed ${\alpha}$-synuclein fibrils in vitro. Taken together, our results suggest that a Tet-Off ${\beta}23$ cell model could serve as a robust platform for screening effective lead compounds inhibiting nuclear or cytosolic protein aggregates. Brain-permeable PCIII or its derivatives could be beneficial for eliminating established protein aggregates.

A case of maxilla implant overdenture using Pekkton telescopic attachment with severe alveolar bone resorption (심한 치조골 소실이 있는 상악 무치악 환자에서 Pekkton telescopic attachment를 이용한 임플란트 피개의치 증례)

  • Park, Ha Eun;Lee, Won Sup;Lee, Cheol Won;Lee, Su Young
    • The Journal of Korean Academy of Prosthodontics
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    • v.57 no.2
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    • pp.189-194
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    • 2019
  • It is necessary to set the correct occlusal plane and to distribute the occlusal force uniformly considering the state of the opposing dentition during the prosthetic of the single edentulous patient with severe alveolar bone resorption. Implant supported overdenture is superior to complete denture in terms of maintenance and stability, and limited implants are used in fully edentulous patients with high alveolar bone resorption. Telescopic attachments using a newly introduced material based on poly-aryl-ether-ketone (PAEK) have the advantages of typical telescopic copping, excellent abrasion resistance, and are lighter and more economical than conventional implant overdentures. In this case, we restored maxillary arch with a implant retained overdenture using the telescopic attachment made of Pekktonand the mandible was restored with fixed implant prosthesis. Through these procedures esthetic aspects and functional outcomes were satisfactorily achieved.