• Title/Summary/Keyword: $\alpha$-CD

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Immune Responses of Mice Intraduodenally Infected with Toxoplasma gondii KI-1 Tachyzoites

  • Shin, Eun-Hee;Chun, Yeoun-Sook;Kim, Won-Hee;Kim, Jae-Lip;Pyo, Kyoung-Ho;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.49 no.2
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    • pp.115-123
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    • 2011
  • Toxoplasma gondii Korean isolate (KI-1) tachyzoites were inoculated intraduodenally to BALB/c mice using a silicon tube, and the course of infection and immune responses of mice were studied. Whereas control mice, that were infected intraperitoneally, died within day 7 post-infection (PI), the intraduodenally infected mice survived until day 9 PI (infection with $1{\times}10^5$ tachyzoites) or day 11 PI (with $1{\times}10^6$ tachyzoites). Based on histopathologic (Giemsa stain) and PCR (B1 gene) studies, it was suggested that tachyzoites, after entering the small intestine, invaded into endothelial cells, divided there, and propagated to other organs. PCR appeared to be more sensitive than histopathology to detect infected organs and tissues. The organisms spread over multiple organs by day 6 PI. However, proliferative responses of splenocytes and mesenteric lymph node (MLN) cells in response to con A or Toxoplasma lysate antigen decreased significantly, suggesting immunosuppression. Splenic $CD4^+$ and $CD8^+$ T-Iymphocytes showed decreases in number until day 9 PI, whereas IFN-${\gamma}$ and IL-10 decreased slightly at day 6 PI and returned to normal levels by day 9 PI. No TNF-${\alpha}$ was detected throughout the experimental period. The results showed that intraduodenal infection with KI-1 tachyzoites was successful but did not elicit significant mucosal immunity in mice and allowed dissemination of T. gondii organisms to systemic organs. The immunosuppression of mice included reduced lymphoproliferative responses to splenocytes and MLN cells to mitogen and low production of cytokines, such as IFN-${\gamma}$, TNF-${\alpha}$, and IL-10, in response to T. gondii infection.

Development of quality of life with WHOQOL-HIV BREF Korean version among HIV patients in Korea (후천성면역결핍환자의 삶의 질 측정을 위한 한국판 WHOQOL-HIV BREF 개발)

  • Lee, Won Kee;Kim, Shin-Woo;Kim, Hye-In;Chang, Hyun-Ha;Lee, Jong-Myung;Kim, Yoon-Joo;Lee, Mi-Young
    • Journal of the Korean Data and Information Science Society
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    • v.25 no.2
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    • pp.337-347
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    • 2014
  • There is no known publication about assessment of quality of life (QOL) in Korean HIV patients. We aimed to assess the QOL of HIV patients. We developed Korean version of the WHOQOL-HIV BREF (short forms of WHOQOL-HIV, 31 questions with 6 domains). Survey data from 220 HIV-positive adults were obtained in 14 centers in South Korea. Male were dominant (202/220, 91.8%). Mean age was $40.6{\pm}12.1$. Mean CD4+ T-cell count was $414.9{\pm}226.6/ml$. Overall of WHOQOL-HIV BREF were $53.2{\pm}14.9$ (perfect score=100) (Cronbach's ${\alpha}$ = 0.942). It is similar score comparing to another country (Portugal: 54.75/100, measured by WHOQOL-HIV). Correlations of WHOQOL-HIV BREF score with patients' subjective QOL and with subjective satisfaction were 0.747 (p <0.01) and 0.651 (p <0.01), respectively. WHOQOL-HIV BREF have internal reliability. There is in need of monitoring for QOL of HIV patients in the clinical practice and trials. This survey tool could be used to assess the effect of intervention. Additionally, comparison across countries would be possible and promising.

Ameliorating Effects of Lactic Acid-fermented Garlic Extracts on Oleic Acid-induced Hepatic Steatosis (유산균 발효 마늘 추출물의 oleic acid로 유도된 비알코올성 지방간에 대한 개선 효과)

  • Lee, Hee-Seop;Lim, Won-Chul;Choi, Ji-Hwi;Yu, Heui-Jong;Kim, Ki-Ho;Lee, Seung-Hyun;Cho, Hong-Yon
    • Korean Journal of Food Science and Technology
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    • v.46 no.6
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    • pp.762-768
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    • 2014
  • In this study, the ameliorating effects of lactic acid-fermented garlic extract (LAFGE) on non-alcoholic fatty liver were investigated using oleic acid-induced steatotic HepG2 cells. The ameliorating mechanism was analyzed by RT-PCR and Western blot. Treatment with 1 mg/mL LAFGE decreased intracellular lipid accumulation approximately 1.5-fold, compared to that achieved with non-fermented garlic extract. LAFGE reduced fatty acid influx into hepatocytes through down-regulation of FAT/CD36 mRNA expression in the steatotic HepG2 cells. $PPAR{\alpha}$ and CPT-1 mRNA expression was significantly up-regulated by LAFGE treatment of HepG2 cells as a consequence of activation of beta oxidation. Additionally, the treatment with 1 mg/mL LAFGE highly down-regulated mRNA expression of SREBP-1c and FAS to 51% and 35%, respectively. LAFGE showed concentration-dependent down-regulation patterns in protein expression of SREBP-1c and FAS, as determined by Western blot. These results suggest that LAFGE treatment improves hepatic steatosis triggered by the imbalance of hepatic lipid metabolism owing to oleic acid treatment.

Immunomodulatory activities of crude polysaccharide fraction separated from Perilla frutescens Britton var. acuta Kudo (자소엽(Perilla frutescens Britton var. acuta Kudo) 조다당류 추출물의 면역활성 효과)

  • Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.49 no.5
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    • pp.559-566
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    • 2017
  • This aim of this study was to examine the immunomodulatory activities of crude polysaccharides from Perilla frutescens Britton var. acuta Kudo (PCP) in mouse bone marrow-derived dendritic cells (BMDC) and splenocytes. The immunomodulatory activity was determined by cell viability, nitric oxide (NO) production, cell surface marker expression (CD 80/86 and MHC class I/II), and cytokine production in BMDC, and cell viability, and cytokine production in splenocytes. Cell proliferation and cytokine production (tumor necrosis factor; TNF-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and IL-12) tested in BMDC were significantly increased by PCP treatment. Additionally, the cell surface markers (CD 80/86, MHC class I/II) were highly increased by PCP treatment. For cytokine production in splenocytes, PCP treatment significantly increased the production of Th 1 cytokines [IL-2 and interferon (IFN)-${\gamma}$], but not Th 2 cytokines (IL-4). Therefore, PCP can induce immune cell activation and is a potential candidate for the development of nutraceuticals to boost the immune system.

Antifreeze Activity of Dimerized Type I Antifreeze Protein Fragments (Type I 결빙방지 단백질 조각 이량체의 결빙방지 활성)

  • Kim, Hak Jun
    • Journal of Life Science
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    • v.27 no.5
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    • pp.584-590
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    • 2017
  • Antifreeze proteins (AFPs) bind to ice crystals and inhibit their growth. AFPs are essential for the survival of organisms living in subzero environments. Type I AFP (AFP37) isolated from winter flounder is an ${\alpha}$-helical peptide of 37 residues long. In this study, we attempted to develop short AFP fragments with higher activity and solubility. We designed and synthesized N-terminal 15 and 21 residue-long AFPs, designated AFP15 and 21. Also dimerized AFP15 and 21, designated dAFP15N and dAFP21N, respectively, were generated through disulfide bonds between peptides containing CGG residues added to the N-terminus of AFP15 and AFP21 (designated AFP15N and 21N). Their helical contents and antifreeze activities were assessed using circular dichroism (CD) spectroscopy and a nanoliter osmometer, respectively. The helical content of AFP15 AFP21, AFP15N, AFP21N, dAFP15N and dAFP21N was 47, 48, 23.8, 28, 49.1, and 52%, respectively compared to that of wild type AFP37; the antifreeze activity was 8.4, 9.3, 0.05, 5.6, 12.1, 11.2% respectively, compared to that of wild type AFP37. Contrary to our anticipation, the dimerized peptides showed almost the same antifreeze activity as their monomeric counterparts. These results indicate that the dimerized peptides behave as monomeric peptides due to the high rotational freedom of disulfide bonds connecting two monomeric peptides. The star-shaped ice crystals generated by the peptides also demonstrated weak interaction between ice and peptides.

A study on the electrodeposition of uranium using a liquid cadmium cathode at 440℃ and 500℃ (440℃와 500℃에서 액체카드뮴음극을 이용한 우라늄 전착에 관한 연구)

  • Yoon, Jong-Ho;Kim, Si-Hyung;Kim, Gha-Young;Kim, Tack-Jin;Ahn, Do-Hee;Paek, Seungwoo
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.11 no.3
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    • pp.199-206
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    • 2013
  • Electrowinning process in pyroprocessing recovers U (uranium) and TRU (Trans Uranium) elements simultaneously from spent fuels using a liquid cadmium cathode (LCC). When the solubility limit of U deposits over 2.35wt% in Cd, U dendrites were formed on the LCC surface during the electrodeposition at $500^{\circ}C$. Due to the high surface area of dendritic U, the deposits were not submerged into the liquid cadmium pool but grow out of the LCC crucible. Since the U dendrites act as a solid cathode, it prevents the co-deposition of U and TRUs. In this study, the electrodeposition of U onto a LCC was carried out at 440 and $500^{\circ}C$ to compare the morphology and component of U deposits. The U deposits at $440^{\circ}C$ have a specific shape and were stacked regularly at the center of the LCC pool, while the U dendrites (i.e., ${\alpha}$-phase) at $500^{\circ}C$ were grow out of the LCC crucible. Through the microscopic observation and XRD analysis, the electrodeposits at $440^{\circ}C$, which have a round shape, were identified as an intermetallic compound such as $UCd_{11}$. It can be concluded that the LCC electrowinning operation at $440^{\circ}C$ achieves the co-recovery of U and TRU without the formation of U dendrites.

Purification and Biological Characterization of Wild-type and Mutants of a Levan Fructotransferase from Microbacterium sp. AL-210 (Microbacterium sp. A-210이 생성하는 Levan fructotransferase의 정제 및 생물학적 특성에 관한 연구)

  • Hwang, Eun-Young;Jeong, Mi-Suk;Cha, Jae-Ho;Jang, Se-Bok
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1218-1225
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    • 2009
  • Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

Anti-Obese Activity of HPJ Extract on High Fat Diet-Induced Obese Mice (고지방 식이로 유도된 비만 쥐에서 HPJ 추출물의 항비만 효과)

  • Yuan, Hai-Dan;Quan, Hai-Yan;Zhang, Ya;Kim, Sung-Jib;Shin, Dae-Hee;Lim, Bang-Ho;Chung, Sung-Hyun
    • YAKHAK HOEJI
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    • v.53 no.5
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    • pp.286-292
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    • 2009
  • In this study, we investigated the anti-obese activity of HPJ extract in C57BL/6J mice. The C57BL/6J mice were randomly divided into five groups: normal control group (Con), high fat diet control group (HFD), treatment groups with HPJ at 125 mg/kg (HPJ125), 250 mg/kg (HPJ250), or 500 mg/kg (HPJ500). To induce an obesity, mice were fed by a high fat diet for 6 weeks, and mice were administered with HPJ extract once a day for 8 weeks. At the end of treatment, we examined the effect of HPJ extract on body weight, plasma lipid, and lipogenic enzymes. HPJ extract was found to lower whole body and epididymal adipose tissue weights and lowered plasma levels of glucose, insulin, triglyceride (TG), total cholesterol (TC), non-esterified fatty acid (NEFA) and leptin, compared to those in HFD group. Histological analyses of the liver and fat tissues of mice treated with HPJ extract revealed significantly decreased number of lipid droplets and decreased size of adipocytes compared to the HFD group. In addition, HPJ extract preserved the morphological integrity of pancreatic islets. To elucidate an action mechanism of HPJ extract, Western blot and RT-PCR were performed using epididymal adipose tissues. HPJ extract up-regulated the levels of phosphorylated adenosine monophosphate-activated protein kinase (AMPK) and its substrate, acetyl-CoA carboxylasse (ACC). HPJ extract also attenuated lipogenic gene expressions of sterol regulatory element-binding protein $1{\alpha}$ (SREBP$1{\alpha}$), fatty acid synthase (FAS), sterol-CoA desaturase 1 (SCD1) and glycerol-3-phosphate acyltransferase (GPAT) in dose-dependent manners. In contrast, expressions of lipolytic genes such as peroxisome proliferator-activated receptor-$\alpha$ (PPAR-${\alpha}$) and CD36, and fatty acid $\beta$-oxidation gene, carnitine palmitoyltransferase-1 (CPT-1) were increased. These results suggest that HPJ extract ameliorates obesity through inhibiting synthesis of lipogenic enzymes as well as stimulating fatty acid oxidation resulting from activation of AMPK, and HPJ extract could be developed as a potential therapeutic agent for obese patients.

Nucleotide Sequence, Structural Investigation and Homology Modeling Studies of a Ca2+-independent α-amylase with Acidic pH-profile

  • Sajedi, Reza Hassan;Taghdir, Majid;Naderi-Manesh, Hossein;Khajeh, Khosro;Ranjbar, Bijan
    • BMB Reports
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    • v.40 no.3
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    • pp.315-324
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    • 2007
  • The novel $\alpha$-amylase purified from locally isolated strain, Bacillus sp. KR-8104, (KRA) (Enzyme Microb Technol; 2005; 36: 666-671) is active in a wide range of pH. The enzyme maximum activity is at pH 4.0 and it retains 90% of activity at pH 3.5. The irreversible thermoinactivation patterns of KRA and the enzyme activity are not changed in the presence and absence of $Ca^{2+}$ and EDTA. Therefore, KRA acts as a $Ca^{2+}$-independent enzyme. Based on circular dichroism (CD) data from thermal unfolding of the enzyme recorded at 222 nm, addition of $Ca^{2+}$ and EDTA similar to its irreversible thermoinactivation, does not influence the thermal denaturation of the enzyme and its Tm. The amino acid sequence of KRA was obtained from the nucleotide sequencing of PCR products of encoding gene. The deduced amino acid sequence of the enzyme revealed a very high sequence homology to Bacillus amyloliquefaciens (BAA) (85% identity, 90% similarity) and Bacillus licheniformis $\alpha$-amylases (BLA) (81% identity, 88% similarity). To elucidate and understand these characteristics of the $\alpha$-amylase, a model of 3D structure of KRA was constructed using the crystal structure of the mutant of BLA as the platform and refined with a molecular dynamics (MD) simulation program. Interestingly enough, there is only one amino acid substitution for KRA in comparison with BLA and BAA in the region involved in the calcium-binding sites. On the other hand, there are many amino acid differences between BLA and KRA at the interface of A and B domains and around the metal triad and active site area. These alterations could have a role in stabilizing the native structure of the loop in the active site cleft and maintenance and stabilization of the putative metal triad-binding site. The amino acid differences at the active site cleft and around the catalytic residues might affect their pKa values and consequently shift its pH profile. In addition, the intrinsic fluorescence intensity of the enzyme at 350 nm does not show considerable change at pH 3.5-7.0.

A Co-inhibitory Molecule, B7-H4, Synergistically Potentiates Oral Tolerance by Inducing CD4+CD25+FoxP3+ T Cells

  • Wen, Lanying;Yang, Sung-Yeun;Choi, Jae-Kyoung;Kim, Young-Hee;Kwon, Eun-Hee;Lee, Hyun-Ji;Jeoung, Hae-Young;Hwang, Du-Hyeon;Hwang, Dong-Jin;Choi, In-Hak
    • IMMUNE NETWORK
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    • v.8 no.1
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    • pp.21-28
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    • 2008
  • Background: A co-inhibitory molecule, B7-H4, is believed to negatively regulate T cell immunity by suppressing T cell proliferation and inhibiting cytokine production. However, the mechanism behind B7-H4-mediated tolerance remains unclear. Methods: Balb/c $(H-2^d)$ mice were fed with dendritic cell line, DC2.4 $(H-2^d)$ every day for 10 days. Meantime, mice were hydrodynamically injected with recombinant plasmid expressing B7-H4 fusion protein (B7-H4.hFc) or hFc via tail vein. One day after last feeding, mice were immunized with allogeneic B6 spleen cells. 14 days following immunization, mice were challenged with B6 spleen cells to ear back and the ear swelling was determined the next day. Subsequently, a mixed lymphocyte reaction (MLR) was also performed and cytokines profiles from the reaction were examined by sandwich ELISA. Frequency of immunosuppressive cell population was assayed with flow cytometry and mRNA for FoxP3 was determined by RT-PCR. Results: Tolerant mice given plasmid expressing B7-H4.hFc showed a significant reduction in ear swelling compared to control mice. In addition, T cells from mice given B7-H4.hFc plasmid revealed a significant hyporesponsiveness of T cells against allogeneic spleen cells and showed a significant decrease in Th1 and Th2 cytokines such as IFN-${\gamma}$, IL-5, and TNF-${\alpha}$. Interestingly, flow cytometric analysis showed that the frequency of CD4+CD25+FoxP3+ Tregs in spleen was increased in tolerant mice given recombinant B7-H4.hFc plasmid compared to control group. Conclusion: Our results demonstrate that B7-H4 synergistically potentiates oral tolerance induced by allogeneic cells by increasing the frequency of FoxP3+ CD4+CD25+ Treg and reducing Th1 and Th2 cytokine production.