• Molecules and Cells
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• 제19권3호
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• pp.398-401
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• 2005

The bacterial enzyme UDP-N-acetylglucosamine enolpyruvyl transferase catalyzes the first committed step of peptidoglycan biosynthesis, i.e., transfer of enolpyruvate from phosphoenolpyruvate to UDP-N-acetyl-glucosamine. We have overexpressed the enzyme from Haemophilus influenzae in Escherichia coli and crystallized it in the apo-form, as well as in a complex with UDP-N-acetylglucosamine and fosfomycin using ammonium sulfate as the precipitant. X-ray diffraction data from a crystal of the apo-form were collected to $2.8{\AA}$ resolution at 293 K. The crystal quality was improved by co-crystallization with UDP-N-acetylglucosamine and fosfomycin. X-ray data to $2.2{\AA}$ have been collected at 100 K from a flash-frozen crystal of the complex. The complex crystals belong to the orthorhombic space group I222 (or $I2_12_12_1$) with unit-cell parameters of a = 63.7, b = 124.5, and $c=126.3{\AA}$. Assuming a monomer of the recombinant enzyme in the crystallographic asymmetric unit, the calculated Matthews parameter ($V_M$) is $2.71{\AA}^3Da^{-1}$ and solvent content is 54.6%.

• 한방재활의학과학회지
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• 제19권4호
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• pp.59-78
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• 2009

Objective : This study was designed to examine the effects of stem bark extracts of Cornus walteri Wanger on the lipid lowering, anti-oxidative activity and concentration of proinflammatory cytokines in hyperlipidemic rat. Methods : Male rats weighing $195.21{\pm}5.85g$ fed high fat diet for 8 weeks and 40 rats(above 400 g) were divided into 4 groups. Each groups were divided into a control group and 3 experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats Basal diet and administered an extract of Cornus walteri Wanger(100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflammatory cytokines, anti-oxidative activity and gene expression. Results : 1. Concentration of plasma free fatty acid, LDL-cholesterol showed a tendency to decrease in Cornus walteri Wanger ext. groups. Concentration of plasma triglyceride, total cholesterol showed a significantly decrement in all Cornus walteri Wanger ext. group than that of control group. HDL-cholesterol showed a significantly increment in the 300 mg/kg Cornus walteri Wanger ext. group. 2. Concentration of liver total cholesterol showed a tendence to decrease in Cornus walteri Wanger ext. groups. Concentration of triglyceride liver showed a significantly decrement in all Cornus walteri Wanger ext. group than that of control group. 3. Concentration of plasma and liver TBARS showed a tendence to decrease in Cornus walteri Wanger ext. groups. The values of GSH-Px, SOD and CAT activity showed a significantly increment in the 300 mg/kg Cornus walteri Wanger ext. group than that of control group. 4. The values of plasma AST and ALT activity showed no significantly different in all treatment groups. 5. Concentration of plasma $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ showed a tendency to decrease in the Cornus walteri Wanger ext. groups. However the concentration of IL-10 in the 200 and 300 mg/kg Cornus walteri Wanger ext. groups showed a significantly increment than that of control group. Concentration of liver $IL-1{\beta}$, $TNF-{\alpha}$ and IL-10 showed no significantly difference in all treatment groups. However concentration of IL-6 in the 200 and 300 mg/kg Cornus walteri Wanger ext. groups showed a significantly decrement than that of control group. 6. In the analysis of RT-PCR, gene expression of $TNF-{\alpha}$, Apo-B, Apo-E and leptin in the Cornus walteri Wanger ext. groups showed a lower expression than that of control group. 7. The ratio of $TNF-{\alpha}$, Apo-E and leptin expression per $\beta$-actin expression in the 200 and 300 mg/kg Cornus walteri Wanger ext. showed a significantly decrement than that of control group. The ratio of Apo-B expression per $\beta$-actin expression in the 300 mg/kg Cornus walteri Wanger ext. showed a significantly decrement than that of control group. Conclusions : According to above results, in lowering lipid effect, antioxidative activity and antiinflammatory effect, the Cornus walteri Wanger ext. gives positive effect.

• Journal of Periodontal and Implant Science
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• 제50권3호
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• pp.159-170
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• 2020

Purpose: Immunization with Porphyromonas gingivalis heat shock protein 60 (PgHSP60) may have an immunoregulatory effect on atherogenesis. The aim of this study was to determine whether nasal immunization with a PgHSP60 peptide could reduce atherosclerotic plaque formation in apolipoprotein E knockout (ApoE KO) mice. Methods: Seven-week-old male ApoE KO mice were assigned to receive a normal diet, a Western diet, a Western diet and challenge with PgHSP60-derived peptide 14 (Pep14) or peptide 19 (Pep19), or a Western diet and immunization with Pep14 or Pep19 before challenge with Pep14 or Pep19. Results: Atherosclerotic plaques were significantly smaller in mice that received a Western diet with Pep14 nasal immunization than in mice that received a Western diet and no Pep14 immunization with or without Pep14 challenge. An immunoblot profile failed to detect serum reactivity to Pep14 in any of the study groups. Stimulation by either Pep14 or Pep19 strongly promoted the induction of CD4⁺CD25⁺ forkhead box P3 (FoxP3)⁺ human regulatory T cells (Tregs) in vitro. However, the expression of mouse splenic CD4⁺CD25⁺FoxP3⁺ Tregs was lower in the Pep14-immunized mice than in the Pep14-challenged or Pep19-immunized mice. Levels of serum interferon gamma (IFN-γ) and transforming growth factor beta were higher and levels of interleukin (IL) 10 were lower in the Pep14-immunized mice than in the other groups. Induction of CD25^- IL-17⁺ T helper 17 (Th17) cells was attenuated in the Pep14-immunized mice. Conclusions: Nasal immunization with Pep14 may be a mechanism for attenuating atherogenesis by promoting the secretion of IFN-γ and/or suppressing Th17-mediated immunity.

• 한국미생물·생명공학회지
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• 제25권2호
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• pp.137-143
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• 1997

Porcine transforming growth factor-$\beta$1 (TGF-$\beta$1) was expressed in Escherichia coli using cDNA of TGF-$\beta$1 and glutathione S-transferase (GST) fusion vector pGEX-1$\lambda$T. An ApoI-Tth111I fragment of cDNA which correspond to the amino acid residues from 123 to 390 of the precursor TGF-$\beta$1 was inserted into EcoRI-Tth111I digested pGEM#-l$\lambda$T and the recombined plasmid was named pGET-12. Gene products from the cloned regions of the recombinant plasmids pGET-12 was not detected in soluble fraction of cell free extract but detected in insoluble fraction. The solubilization of insoluble gene product was achieved by the treatment of N-laurylsarcosine. Molecular weight of partially purified proteins determined by electrophoresis was same as expected from cloned fragment. The ELISA test results of the purified proteins showed that immunologically detectable epitope was preserved in recombinant protein.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2016년도 제50회 동계 정기학술대회 초록집
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• pp.372.1-372.1
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• 2016

Recently, there are a lot of diseases all around the world. Out of them, Atherosclerosis (AS) is the most common cause of stroke, cardiovascular mortality, and myocardial infarction. The macrophage-derived foam cell, which is formed by oxidized low-density lipoprotein (oxLDL), is the crucial marker for AS. In this study, we report a label-free capacitance imaging technique with multi-electrode array (MEA). The lipid-rich aorta arch lesions, which are derived from an apolipoprotein-E receptor-deficient (apoE-/-) mouse, exhibit higher capacitance than the lipid-free aorta arch, allowing the capacitance imaging of lipid region in atherosclerosis. To improve the contacts between MEA and tissue, polypyrrole(PPy)-coated multi walled carbon nanotubes (MWNTs) multi electrode array (PPy-MWNTs-MEA) was fabricated. Compared to TiN-MEA, PPy-MWNTs-MEA yielded lower contact impedance and better capacitance images. In addition, we have also developed a flexible MEA using single walled carbon nanotubes on a PET substrate. The lipid region could be discriminated in the capacitance images of the lipid-rich aorta arch lesions measured using flexible MEA, demonstrating a feasibility of in vivo applications.

• Toxicological Research
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• 제17권
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• pp.289-292
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• 2001

Gene targeting allows precise, predetermined changes to be made in a chosen gene in the mouse genome. To date, targeting has been used most often for generation of animals completely lacking the product of a gene of interest. Models of essential hypertension have been produced by mutated genes relating renin angiotensin system. The most significant contribution to understanding the genetic etiology of essential hypertension is probably the demonstration that discrete alterations in the expression of a variety of different genes can individually cause changes in the blood pressures of mice, even when the mice have all their compensatory mechanisms intact. These effects are readily detected in animals having moderate decreases in gene function due to heterozygosity for gene disruptions or modest increases due to gene duplication. As a species the mouse is highly resistant to atherosclerosis. However. through induced mutations it has been possible to develop lines oj mice that are deficient in apolipoprotein E, a ligand important in lipoprotein clearance, develop atherosclerotic lesions resembling those observed in humans. The atherosclerotic lesions in apoE-deficient mice have been well characterized, and they resemble human lesions in their sites of predilection and progression to the fibroproliferative stage. Other promising models are mice that are deficient in the low-density lipoprotein receptor. Considerable work still remains to be done in dissecting out in a rigorous manner the effects of alterations in single genes on the induction or progression of atherosclerosis and on the control of blood pressures. Perhaps even more exciting is the opportunity now becoming available to breed animals in which the effects oj precise differences in more than one gene can be studied in combination.

• Journal of Microbiology and Biotechnology
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• 제19권10호
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• pp.1109-1121
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• 2009

In an attempt to discover novel biomarker proteins in type 2 diabetes prognosis, we investigated the influence of hypoglycemic extracellular polysaccharides (EPS) obtained from the macrofungus Tremella fuciformis on the differential levels of plasma proteins in ob/ob mice using two-dimensional gel electrophoresis (2-DE). The 2-DE analysis demonstrated that 92 spots from about 900 visualized spots were differentially regulated, of which 40 spots were identified as principal diabetes-associated proteins. By comparing control with EPS-fed mice, we found that at least six proteins were significantly altered in ob/ob mice, including Apo A-I, IV, C-III, E, retinol-binding protein 4, and transferrin, and their levels were interestingly normalized after EPS treatment. Western blot analysis revealed that the altered levels of the two regulatory molecules highlighted in diabetes and obesity (e.g., resistin and adiponectin) were also normalized in response to EPS. The Mouse Diabetes PCR Array profiles showed that the expression of 84 genes related to the onset, development, and progression of diabetes were significantly downregulated in liver, adipocyte, and muscle of ob/ob mice. EPS might act as a potent regulator of gene expression for a wide variety of genes in ob/ob mice, particularly in obesity, insulin resistance, and complications from diabetes mellitus.

• The Korean Journal of Physiology and Pharmacology
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• 제22권2호
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• pp.127-134
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• 2018

Myofibrillogenesis regulator-1 (MR-1) is a novel protein involved in cellular proliferation, migration, inflammatory reaction and signal transduction. However, little information is available on the relationship between MR-1 expression and the progression of atherosclerosis. Here we report atheroprotective effects of silencing MR-1 in a model of Ang II-accelerated atherosclerosis, characterized by suppression focal adhesion kinase (FAK) and nuclear factor kappaB ($NF-{\kappa}B$) signaling pathway, and atherosclerotic lesion macrophage content. In this model, administration of the siRNA-MR-1 substantially attenuated Ang II-accelerated atherosclerosis with stabilization of atherosclerotic plaques and inhibited FAK, Akt, mammalian target of rapamycin (mTOR) and NF-kB activation, which was associated with suppression of inflammatory factor and atherogenic gene expression in the artery. In vitro studies demonstrated similar changes in Ang II-treated vascular smooth muscle cells (VSMCs) and macrophages: siRNA-MR-1 inhibited the expression levels of proinflammatory factor. These studies uncover crucial proinflammatory mechanisms of Ang II and highlight actions of silencing MR-1 to inhibit Ang II signaling, which is atheroprotective.

• 한국식품영양과학회지
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• 제43권4호
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• pp.491-497
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• 2014

본 연구는 고콜레스테롤 식이를 급여한 마우스의 지질대사에 알긴산 올리고당이 미치는 영향을 조사하였다. 실험동물은 4주령 된 수컷 apoE 마우스를 사용하였으며, 정상군, 고콜레스테롤 식이대조군, 알긴산 올리고당 5%와 10% 첨가군으로 나누어 6주간 급여하였다. 실험식이는 AIN-93G 식이를 기본으로 하여 콜레스테롤(1%, w/w)을 혼합하였으며, 알긴산 올리고당을 각각 5%와 10% 수준으로 혼합하였다. 실험기간 동안 증체량은 고콜레스테롤 식이대조군에 비해 알긴산 올리고당 10% 첨가군이 약 7.6% 유의적으로 감소하였다(P<0.05). 장기조직 중 신장 주변지방과 부고환지방의 무게는 고콜레스테롤 식이대조군이 정상군에 비해 각각 42.6%, 27.6% 유의적으로 증가하였다(P<0.05). 반면 알긴산 올리고당 첨가군은 신장 주변지방과 부고환지방 무게가 고콜레스테롤 식이대조군보다 2배 이상 유의적으로 감소하였다(P<0.05). 고콜레스테롤 섭취는 혈청 중 총 콜레스테롤, 중성지방, LDL-콜레스테롤 수준을 정상군보다 각각 16%, 12%, 47.1% 유의적으로 증가시켰으며, 반면에 알긴산 올리고당 10%를 섭취한 경우 고콜레스테롤 식이대조군보다 각각 57.5%, 51.4%, 82.9% 유의적으로 감소시켰다 (P<0.05). 혈청 HDL-콜레스테롤 함량은 실험군간 유의적 차이는 크게 나타나지 않았으나, 전체 콜레스테롤에 대한 HDL-콜레스테롤 비율에서 알긴산 올리고당 첨가군이 유의적으로 높았다(P<0.05). 간 조직 내 지질 수준은 고콜레스테롤을 섭취했을 경우 정상군에 비해 유의적으로 증가하였으나, 알긴산 올리고당 10%를 섭취했을 때 고콜레스테롤 식이대조군보다 총 콜레스테롤 함량은 3배 이상, 중성지방 함량은 33.5% 유의적으로 감소하였다(P<0.05). 고콜레스테롤 식이군에서 나타난 분변 내 총 콜레스테롤 함량은 알긴산 올리고당 10% 첨가군에서 정상군보다 21% 유의적으로 증가하였다(P<0.05). 이상의 실험 결과 고콜레스테롤 식이와 알긴산 올리고당을 함께 섭취한 마우스는 체중 감량과 장기 내 지방의 중량을 감소시켜 비만 예방효과가 있고, 혈액, 간지질 및 분변 지질 수준의 개선 효과가 높아 지질대사와 관련된 만성질환 예방에 효과가 있을 것으로 기대된다.

• Molecules and Cells
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• 제27권1호
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• pp.99-103
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• 2009

Ribose-5-phosphate isomerase A (RpiA) plays an important role in interconverting between ribose-5-phosphate (R5P) and ribulose-5-phosphate in the pentose phosphate pathway and the Calvin cycle. We have determined the crystal structures of the open form RpiA from Vibrio vulnificus YJ106 (VvRpiA) in complex with the R5P and the closed form with arabinose-5-phosphate (A5P) in parallel with the apo VvRpiA at $2.0{\AA}$ resolution. VvRpiA is highly similar to Escherichia coli RpiA, and the VvRpiA-R5P complex strongly resembles the E. coli RpiA-A5P complex. Interestingly, unlike the E. coli RpiA-A5P complex, the position of A5P in the VvRpiA-A5P complex reveals a different position than the R5P binding mode. VvRpiA-A5P has a sugar ring inside the binding pocket and a phosphate group outside the binding pocket: By contrast, the sugar ring of A5P interacts with the Asp4, Lys7, Ser30, Asp118, and Lys121 residues; the phosphate group of A5P interacts with two water molecules, W51 and W82.