• Title/Summary/Keyword: Outer membrane

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The Effects of Puerariae Radix Catechins Administration on Liver Function in Carbon Tetrachloride-Treated Rats (사염화탄소를 투여한 흰쥐에 있어서의 간 기능에 미치는 칡 카테킨의 효과)

  • 이치호;한석현;김종배;민상기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.5
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    • pp.713-719
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    • 1995
  • These studies were carried out to investigate the effects of Puerariae radix catechins(PRC) administration on the biochemical parameters of liver function in liver of carbon tetrachloride(CCl4)-treated rats. Thirty six healthy Sprague-Dawley rats weighing about 120g were used for this experiment and divided intot he following 3 groups : normal control group(NCON), $CCl_4$ control group(CCON), PRC treated group(PRC). Fifty percent $CCl_4$ in oil was administered(I.P.) by 2ml per kg body weight two times a week for 3 weeks. PRC treated groups were administered orally at the leaves of 1% per day in distilled water for 8 weeks. Lipid hydroperoxides were analyzed by using chemiluminescence-high performance liquid chromatography(CL-HPLC) method as a phosphatidylcholine hydroperoxide value(PCOOH) in liver tissues. $CCl_4$ treatment significantly(p<0.05) resulted in an increase in GPT & GOT activities and liver hydroperoxide values comparing with those of the untreated control, while administration of PRC to the $CCl_4-treated$ rats significantly(p<0.001) decreased GPT & GOT activities and liver hydroperoxide value. Their ultrastructual changes of hepatocellular organelles were shown to clarify the morphologic nature of protective effects of PRC on hepatocytic injuries. $CCl_4$ treatment observed to change the ultrastructual nature of outer membrane of hepatocytes. However, the hepatic changes on PRC treatment to $CCl_4$ group was not found. PRC administration may inhibit the formatiion of liver lipid hydroperoxides in vivo and were very effective in recovering the liver function in $CCl_4-treated$ rats.

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Extraction and verification of highly immunologenic antigen for diagnosis of bovine brucellosis (소 브루셀라병 진단용 고면역원성 항원의 추출과 검증)

  • Bae, Jae-Hyung;Jo, Sang-Rae;Jeong, Eun-Hui;Jang, Eun-Hui;Kim, Seong-Eun;Kwon, Hui-Nyeong;Park, Dong-Yeop;Lee, Kuk-Cheon
    • Korean Journal of Veterinary Service
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    • v.33 no.2
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    • pp.135-141
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    • 2010
  • Bovine brucellosis, an important zoonosis, is diagnosed with serological tests such as the RBT, TAT using inactivated whole bacterial cells or bacterial lipopolysaccharide (LPS) antigen in Korea. However, a strong cross-reaction between Brucella spp. and Yersinia enterocolitica O9 in these tests has seriously complicated the diagnosis of animal brucellosis because Brucella spp. shares common antigenic determinants with Y. enterocolitica O9 in the smooth LPS region. In this study, Brucella-field strains were isolated from Brucellapositive Hanwoo in Kimhae, Korea and outer membrane protein (omp) which has low cross-reaction with Y. enterocolitica O9 and high immunogenicity was extracted from the field strains Then we compared ELISA using the extract with RBT-TAT. Fifteen field strains were isolated from 47 supra-mammary-lymph nodes, which were collected from 18 farms. Isolation rate was 32%. Brucella-specific antigen was identified by performing SDS-PAGE or Western blotting on extracted omp with at 0.5% n-lauroylsarcosine One hundred and ninety-two serum-samples were used in the experiment: 142 negative and 50 positive samples verified by RBT-TAT. According to ELISA results, 127 samples were negative and 15 appeared positive among 142 negatives by RBT-TAT, while 42 samples were positive and 8 were negative among 50 positives by RBT-TAT. Therefore, it showed 89.4% of specificity and 84% of sensi-tivity. Through the current experiments, we could set up an ELISA based on the omp which has low cross-reaction and high immunogenicity and concluded that the omp could be a good material for accurate diagnosis of bovine brucellosis.

Enhanced Extraction of Bioactive Compounds from Bee Pollen by Wet-grinding Technology (벌 화분에서 습식 나노화 공정에 의한 유효성분의 추출)

  • Choi, Yun-Sik;Suh, Hwa-Jin;Chung, Il Kyung
    • Journal of Life Science
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    • v.26 no.6
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    • pp.651-656
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    • 2016
  • Bee pollen is produced by honeybees and is considered one of the most balanced and nourishing nutritional supplements available. Historically, bee pollen has been prescribed for its healing properties and consumed for its high-energy supply. Recent research has provided evidence that bee pollen has diverse biological activities, such as anti-oxidant, anti-inflammatory, anti-bacterial, and even anti-cancer effects. However, the outer membrane of the pollen grain, exine, is highly resistant to most acidic solutions, high pressure, and even digestive enzymes, and the resulting low bioavailability limits its nutritional and clinical applications. This study applied a wet-grinding method to destroy the exine effectively, and it then examined the pollen's enhanced biological activity. First, microscopic observations provided strong evidence that wet grinding destroyed the exine time-dependently. In addition, the content of polyphenols, well-known ingredients of bee pollen and used as internal standards for the quality control of commercial pollen preparations, increased up to 11-fold with wet grinding. Further, the anti-oxidant activity demonstrated on the ABTS anti-oxidant assay, as well as the DPPH radical scavenging assay, was also dramatically increased. Together, the results presented here support a new technology by which bee pollen can be used as a resource for medical, nutritional, and cosmetic applications.

The 18-kDa Translocator Protein Inhibits Vascular Cell Adhesion Molecule-1 Expression via Inhibition of Mitochondrial Reactive Oxygen Species

  • Joo, Hee Kyoung;Lee, Yu Ran;Kang, Gun;Choi, Sunga;Kim, Cuk-Seong;Ryoo, Sungwoo;Park, Jin Bong;Jeon, Byeong Hwa
    • Molecules and Cells
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    • v.38 no.12
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    • pp.1064-1070
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    • 2015
  • Translocator protein 18 kDa (TSPO) is a mitochondrial outer membrane protein and is abundantly expressed in a variety of organ and tissues. To date, the functional role of TSPO on vascular endothelial cell activation has yet to be fully elucidated. In the present study, the phorbol 12-myristate 13-acetate (PMA, 250 nM), an activator of protein kinase C (PKC), was used to induce vascular endothelial activation. Adenoviral TSPO overexpression (10-100 MOI) inhibited PMA-induced vascular cell adhesion molecule-1 (VCAM-1) and intracellular cell adhesion molecule-1 (ICAM-1) expression in a dose dependent manner. PMA-induced VCAM-1 expressions were inhibited by Mito-TEMPO ($0.1-0.5{\mu}m$), a specific mitochondrial antioxidants, and cyclosporin A ($1-5{\mu}m$), a mitochondrial permeability transition pore inhibitor, implying on an important role of mitochondrial reactive oxygen species (ROS) on the endothelial activation. Moreover, adenoviral TSPO overexpression inhibited mitochondrial ROS production and manganese superoxide dismutase expression. On contrasts, gene silencing of TSPO with siRNA increased PMA-induced VCAM-1 expression and mitochondrial ROS production. Midazolam ($1-50{\mu}m$), TSPO ligands, inhibited PMA-induced VCAM-1 and mitochondrial ROS production in endothelial cells. These results suggest that mitochondrial TSPO can inhibit PMA-induced endothelial inflammation via suppression of VCAM-1 and mitochondrial ROS production in endothelial cells.

Helicobacter pylori Chaperone-Like Protein CagT Plays an Essential Role in the Translocation of CagA into Host Cells

  • Ding, Honglei;Zeng, Hao;Huang, Linping;Dong, Yandong;Duan, Yijun;Mao, Xuhu;Guo, Gang;Zou, Quanming
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1343-1349
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    • 2012
  • Most of the Helicobacter pylori strains containing the cag pathogenicity island (PAI) have been associated with more severe gastric disease in infected humans. The cag PAI is composed of 27 proteins, and some of the components are required for CagA translocation into host cells as well as induction of proinflammatory cytokines, such as interleukin-8 (IL-8); however, the exact function of most of the components remains unknown or poorly characterized. In this study, we demonstrated that CagT (HP0532), which is an essential structural component of the cag PAI apparatus, plays an important role in the translocation of CagA into host epithelial cells. In addition to being located on the bacterial surface, CagT is also partially localized in the inner membrane, where it acts as a chaperone-like protein and promotes CagA translocation. However, CagT secretion was not detected by immunoprecipitation analysis of cell culture supernatants. Meanwhile, CagT was related to the introduction of IL-8 of the host cell. These results suggest that CagT is expressed on both the inner and outer bacterial membranes, where it serves as a unique type IV secretion system component that is involved in CagA secretion and cag PAI apparatus assembly.

Transmission electron microscopic ultastructure of the tegument of Fibricola seoulenis (Fibricola seouenis 표피의 투과전자현미경적 미세구조)

  • 손운목;이순형
    • Parasites, Hosts and Diseases
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    • v.31 no.4
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    • pp.301-314
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    • 1993
  • An electron microscopic study was performed to observe the ultrastructure of the tegument of U seoulensis. The outer surface of the tegument was covered with a tnlaminated plasma membrane. The electron-dense cytoplasmic layer was $2.5{\;}\mu\textrm{m}$ wide In the anterior portion and contained numerous vacuoles, mitochondriae and granular materials in its matrix. The basement layer was 330 nm wade or so, and Its numerous extensions protruded into the cytoplasmic layer. The sensory organ was composed of a small vesicle of $1.7{\;}{\times}{\;}1.1{\;}\mu\textrm{m}$ in dimensions, which possessed a cilium of $1.2{\;}{\times}{\;}0.19{\;}\mu\textrm{m}$ in size. The pharynx was composed of the epithelial layer of about $0.5{\;}\mu\textrm{m}$ wide, well developed muscle layer and basement layer. The tegument of the oral sucker was composed of a cytoplasmic layer of $0.4-0.5{\;}\mu\textrm{m}$ width, a narrow basement layer, a well developed muscle layer and tegumental cells. Some kinds of secretory granules that seemed to be originated from the cells of the oral sucker were observed In the parenchymal portions of the adjacent cells. The tribocytic organ consisted of numerous microvilli. The microvilli were 5 nm wide and heptalaminated. Two types of secretory granules originated from the gland cells of tribocytic organ were observed In the tegument and parenchyme. The tegumental cells were irregular in shape, and of which nuclei were multifarious.

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Role of FAK Phosphorylation in Cobalt Chloride-Induced Epithelial-to-Mesenchymal-Like Transition (Cobalt chloride에 의해 유도되는 상피-중간엽 이행에서의 국소부착 단백질의 인산화의 역할 규명)

  • Nam, Ju-Ock
    • Journal of Life Science
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    • v.21 no.2
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    • pp.286-291
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    • 2011
  • Hypoxia is a common condition found in a wide range of solid tumors and is often associated with metastasis and poor clinical outcomes. In the present study, we found that HIF-$1{\alpha}$ was induced by cobalt chloride (500 ${\mu}M$) treatment on human lung cancer cells, A549 and H460, for 24 hr. However, cobalt chloride (500 ${\mu}M$) did not affect cell proliferation of A549 and H460 in 48 hr. Cobalt chloride (500 ${\mu}M$) additionally induced epithelial-to-mesenchymal-like transition (EMT) such as reduced E-cadherin expression and increased ${\alpha}$-SMA expression. These results were confirmed by immunofluorecence experiment in H460 cells. E-cadherin was localized on the outer cell membrane. However, when the cells were treated with 500 ${\mu}M$ cobalt chloride for 24 hr, diffuse E-cadherin staining was observed, characteristic of a migratory mesenchymal phenotype. We also found that cobalt chloride induced integrin ${\beta}3$ expression and FAK phosphorylation in human lung cancer cells using western blotting and FACS anlaysis. Our data suggest that integrin ${\beta}3$-induced FAK phosphorylation may be developed into target molecules for blocking tumor metastasis.

Reproductive Cycle of the Goldeye Rockfish, Sebastes thompsoni (Teleostei: Scorpaenidae) (불볼락(Sebastes thompsoni)의 생식주기)

  • LEE Jung Sick;AN Cheul Min;HUH Sung-Hoi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.1
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    • pp.8-16
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    • 1998
  • Sexual maturation and reproductive cycle of the goldeye rockfish, Sebastes thompsoni were investigated under photomicroscopy. Samples were collected monthly in the coastal water of Samcheonpo ($34^{\circ}55'N$ ), Korea from November 1995 to October 1996, The ovary consists of several ovarian lamellae originated from ovarian outer membrane. Oogonia which are originated from the inner surface of the ovarian lamella protrude to the ovarian cavity in oocyte stage, and they ave suspended by the egg stalk. The testis is seminiferous tubule type in internal structure. Seminiferous tubule consists of many testicular cysts which contain numerous germ cells in same developmental stage. Biological minimum size of female and male were 19.5 cm and 21.5 cm in total length, respectively. Gonadosomatic index (GSI) of female was the highest (9.56) in March and the lowest (0.15) in August. GSI of male was the highest (0.25) in February and the lowest (0.04) in July. Reproductive cycle was classified into the following successive stages: in female, growing (October and November), maturation ( $December\~February$), gestation (March), parturition and recovery ($April\~June$) and resting ($July\~September$), and in male, growing ($September\~November$), maturation ( December and January), ripe and copulation ( February and March) and degeneration and resting ($April\~August$).

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Planar microchip-based lactate biosensor (평면 소자형 락테이트 바이오센서)

  • Ha, Jeonghan;Huh, Hwang;Kang, Tae Young;Lee, Yong Seok;Yoon, Soon Ho;Shin, Jungwon;Nam, Hakhyun;Cha, Geun Sig
    • Analytical Science and Technology
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    • v.19 no.6
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    • pp.482-489
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    • 2006
  • Two electrode-based lactate biosensor was prepared by immobilizing lactate oxidase (LOD) obtained from pediococcus species in a poly(vinyl alcohol). Hydrogen peroxide ($H_2O_2$) produced by the reaction of lactate and LOD was detected on the Pt-black that was electrochemically deposited on the Au electrode. Sensors fabricated with Pt-black deposited Au electrode provided a high current of $H_2O_2$ oxidation at a substantially lowered applied potential (+300 mV vs. Ag/AgCl), resulting in reduced interferences from easily oxidizable species such as ascorbic acid, acetaminophen, and uric acid. An outer membrane is formulated by adjusting water uptake of hydrophilic polyurethane (HPU). The sensor performance was evaluated in vitro with both flow-through arrangement and static mode. The sensor showed a linear range from 0.1 mM to about 9.0 mM in 0.05 M phosphate buffer (pH 7.6) containing 0.05 M NaCl. Storing the sensors prepared in this work at $4^{\circ}C$ buffer solution while not in use, they provided same electrochemical performance for more than 25 days.

Immunocytochemical Localization of Parvalbumin and Calbindin-D 28K in Monkey Dorsal Lateral Geniculate Nucleus (원숭이 외측슬상체배측핵에서 칼슘결합단백 Parvalbumin과 Calbindin-D 28K의 분포)

  • Ko, Seung-Hee;Bae, Choon-Sang;Park, Sung-Sik
    • Applied Microscopy
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    • v.24 no.4
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    • pp.61-77
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    • 1994
  • The calcium-binding proteins (CaBP), parvalbumin (PV) and calbindin-D 28K (calbindin) are particularly abundant and specific in their distribution, and present in different subsets of neurons in many brain regions. Although their physiological roles in the neurons have not been elucidated, they are valuable markers of neuronal subpopulations for anatomical and developmental studies. This study is designed to characterize dorsal lateral geniculate nucleus (dLGN) neurons and axon terminals in terms of differential expression of immunoreactivity (IR) for two well-known CaBPs, PV and calbindin. The experiments were carried out on 6 adult monkeys. Monkeys were perfused under deep Nembutal anesthesia with 2% paraformaldehyde and 0.2% glutaraldehyde in 0.1M phosphate buffer. After removal, the brains were postfixed for 6-8 hr in 2% paraformaldehyde at $4^{\circ}C$ and infiltrated with 30% sucrose at $4^{\circ}C$. Thereafter, they were frozen in dry ice. Serial sections of the thalamus, at $20{\mu}m$, were made in the frontal plane with a sliding microtome. The sections were stained for PV and calbindin with indirect immunocytochemical methods. For electron microscopy, after infiltration with 30% sucrose the blocks of thalamus were serially sectioned at $50{\mu}m$ with a Vibratome in the coronal plane and stained immediately by indirect ABC methods without Triton X-100 in incubation medium. Stained sections were postfixed in 0.2% osmium tetroxide, dehydrated and flat-embedded in Spurr resin. The block was then trimmed to contain only a selected lamina or interlaminar space. The dLGN proper showed strong PV IR in fibers in all laminae and interlaminar zones. Particularly dense staining was noted in layers 1 and 2 that contain many stained fibers from optic tract. Neuronal cell body stained with PV was concentrated only in the laminae. In these laminae staining was moderate in cell bodies of all large and medium-sized neurons, and was strong in cell bodies of some small neurons together with their processes. Calbindin IR was marked in the neuronal cell body and neuropil in the S layers and interlaminar zones whereas moderate in the neuropil throughout the nucleus. Regional difference in distribution of PV and calbindin IR cell is distinct; the former is only in the laminae and the latter in both the S layer and interlaminar space. The CaBP-IR elements were confined to about $10{\mu}m$ in depth of Vibratome section. The IR product for CaBP was mainly associated with synaptic vesicle, pre- and post-synaptic membrane, and outer mitochondrial membrane and along microtubule. PV-IR was noted in various neuronal elements such as neuronal soma, dendrite, RLP, F, PSD and some myelinated or unmyelinated axons, and was not seen in the RSD and glial cells. Only a few neuronal components in dLGN was IR for calbindin and its reaction product was less dense than that of PV, and scattered throughout cytoplasm of soma of some relay neurons, and was also persent in some dendrite, myelinated axons and RLP. The RSD, F, PSD and glial elements were always non-IR for calbindin. Calbindin labelled RLP were presynaptic to unlabeled dendrite or dendritic spine and PSD. Calbindin-labeled dendrite of various sizes were always postsynaptic to unlabeled RSD, RLP or F. From this study it is suggested that dLGN cells of different functional systems and their differential projection to the visual cortex can be distinguished by differential expression of PV and calbindin.

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