• Title/Summary/Keyword: NO dimer

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Alkaline Sizing of Mechanical Pulp

  • Kim, Bong-Yong;Akira Isogai
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.32 no.5
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    • pp.1-7
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    • 2000
  • Alkaline Sizing behavior and mechanism of handsheets, which were prepared from thermomechanical pulp (TMP) with alkylketene dimer (AKD), were studied in terms of the conditions of the handsheet-making. AKD content in the TMP handsheets was increased with increasing of AKD addition level and the addition of a polyamideamine-epichlorohydrin resin (PAE) clearly enhanced AKD retention as well as the resultant sizing performance of TMP handsheets. Although drying of the AKD sized TMP webs at $20^{\circ}C$ led to no or quite low sizing level, but TMP handsheets sized with AKD had higher sizing degrees with increasing of the temperature of heat treatment. Scanning electron microscopic observations of the AKD-sized TMP handsheets showed that AKD emulsion particles were present on pulp fiber surfaces independently without coagulation in the TMP handsheets dried at $20^{\circ}C$. Heat treatment of the AKD-sized handsheets resulted in disappearance of the AKD emulsion particles because of their melting and spreading. The addition of calcium carbonate filler to the TMP suspensions did not influence on AKD content in the TMP handsheets. Nevertheless, their sizing degrees clearly increased by the addition of $CaCO_3$filler. Probably, AKD molecules adsorbed on the $CaCO_3$filler particles contribute to the enhancement of sizing performance. Thus, AKD can give sizing features effectively to the TMP handsheets, when they are made under suitable conditions.

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DNA Sequencing and Expression of the Circumsporozoite Protein of Plasmodium vivax Korean Isolate in Escherichia coli

  • Lee, Hyeong-Woo;Lee, Jong-Soo;Lee, Won-Ja;Lee, Ho-Sa
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.234-242
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    • 1999
  • To obtain the recombinant circumsporozoite (CS) protein for the diagnosis of patients and seroepidemiology of Plasmodium vivax malaria which have been prevalent in northern part of Kyonggido, the CS protein gene was amplified by the polymerase chain reaction (PCR) from genomic DNA of the Korean vivax malaria patient. The gene consists of 1,123 nucleotides except signal peptide sequences and had an uninterrupted reading frame encoding a protein of 374 amino acids with a central region of 20 tandem repeats of the nonapeptide. The CS protein gene was expressed in Escherichia coli and purified, the molecular weight of recombinant CS protein was about 44 kDa (monomer) under denaturing purification and about 65 kDa (dimer) under native purification by SDS-PAGE. The purified recombinant CS protein which has antigenicity to malaria patients in Western blot analysis and Enzyme-linked immunosorbent assay, reacted only with the serum of P. vivax (PV210) infected malaria patients with no cross reaction to the P. falciparum malaria patient. The recombinant CS protein purified in this study will serve as a useful antigen to support the diagnosis of malaria patients and seroepidemiology.

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Crystal Structure of β-Carbonic Anhydrase CafA from the Fungal Pathogen Aspergillus fumigatus

  • Kim, Subin;Yeon, Jungyoon;Sung, Jongmin;Jin, Mi Sun
    • Molecules and Cells
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    • v.43 no.9
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    • pp.831-840
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    • 2020
  • The β-class of carbonic anhydrases (β-CAs) are zinc metalloenzymes widely distributed in the fungal kingdom that play essential roles in growth, survival, differentiation, and virulence by catalyzing the reversible interconversion of carbon dioxide (CO2) and bicarbonate (HCO3-). Herein, we report the biochemical and crystallographic characterization of the β-CA CafA from the fungal pathogen Aspergillus fumigatus, the main causative agent of invasive aspergillosis. CafA exhibited apparent in vitro CO2 hydration activity in neutral to weak alkaline conditions, but little activity at acidic pH. The high-resolution crystal structure of CafA revealed a tetramer comprising a dimer of dimers, in which the catalytic zinc ion is tetrahedrally coordinated by three conserved residues (C119, H175, C178) and an acetate anion presumably acquired from the crystallization solution, indicating a freely accessible "open" conformation. Furthermore, knowledge of the structure of CafA in complex with the potent inhibitor acetazolamide, together with its functional intolerance of nitrate (NO3-) ions, could be exploited to develop new antifungal agents for the treatment of invasive aspergillosis.

Activation of ATM/Akt/CREB/eNOS Signaling Axis by Aphidicolin Increases NO Production and Vessel Relaxation in Endothelial Cells and Rat Aortas

  • Park, Jung-Hyun;Cho, Du-Hyong;Hwang, Yun-Jin;Lee, Jee Young;Lee, Hyeon-Ju;Jo, Inho
    • Biomolecules & Therapeutics
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    • v.28 no.6
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    • pp.549-560
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    • 2020
  • Although DNA damage responses (DDRs) are reported to be involved in nitric oxide (NO) production in response to genotoxic stresses, the precise mechanism of DDR-mediated NO production has not been fully understood. Using a genotoxic agent aphidicolin, we investigated how DDRs regulate NO production in bovine aortic endothelial cells. Prolonged (over 24 h) treatment with aphidicolin increased NO production and endothelial NO synthase (eNOS) protein expression, which was accompanied by increased eNOS dimer/monomer ratio, tetrahydrobiopterin levels, and eNOS mRNA expression. A promoter assay using 5'-serially deleted eNOS promoters revealed that Tax-responsive element site, located at -962 to -873 of the eNOS promoter, was responsible for aphidicolin-stimulated eNOS gene expression. Aphidicolin increased CREB activity and ectopic expression of dominant-negative inhibitor of CREB, A-CREB, repressed the stimulatory effects of aphidicolin on eNOS gene expression and its promoter activity. Co-treatment with LY294002 decreased the aphidicolin-stimulated increase in p-CREB-Ser133 level, eNOS expression, and NO production. Furthermore, ectopic expression of dominant-negative Akt construct attenuated aphidicolin-stimulated NO production. Aphidicolin increased p-ATM-Ser1981 and the knockdown of ATM using siRNA attenuated all stimulatory effects of aphidicolin on p-Akt-Ser473, p-CREB-Ser133, eNOS expression, and NO production. Additionally, these stimulatory effects of aphidicolin were similarly observed in human umbilical vein endothelial cells. Lastly, aphidicolin increased acetylcholine-induced vessel relaxation in rat aortas, which was accompanied by increased p-ATM-Ser1981, p-Akt-Ser473, p-CREB-Ser133, and eNOS expression. In conclusion, our results demonstrate that in response to aphidicolin, activation of ATM/Akt/CREB/eNOS signaling cascade mediates increase of NO production and vessel relaxation in endothelial cells and rat aortas.

Isolation of Polyphenol Compounds from the Leaves of Korean Persimmon (Diospyrus kaki L. Folium) (한국산 감잎로부터 Polyphenol계 생리활성물질 분리)

  • An, Bong-Jeun;Bae, Man-Jong;Choi, Hee-Jin;Zhang, Yun-Bin;Sung, Tae-Soo;Choi, Cheong
    • Applied Biological Chemistry
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    • v.45 no.4
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    • pp.212-217
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    • 2002
  • We purified polyphenols from persimmon leaf and tested their biological activity. The 60% acetone extract was lyophilized and applied to test enzyme inhibition of glucosyltransferase and tyrosinase. GTase was 82.4% inhibited at $1.8{\times}10^{-1}$ mg/ml and tyrosinase 21.7% inhibited at 0.8 mg/ml. The acetone extract was fractionated into F-1, 2, 3, 4, 5 by Sephadex Q-50 gel filtration and the fraction-1 and 2 showed higher enzyme inhibition activity than the other fractions. To the Proteinase K treatment and autoclaving of the two fractions had no effect on the enzyme activity, but these results suggested that active fraction was not protein but phenol ring completed compounds. By Sephadex LH-20, MCI-gel and Bondapak $C_{18}$ column chromatographies, compouds 1, 2, 3 and 4 from F-1 fraction, compounds 5 and 6 from F-2 fraction and compounds 7 , 8 from F-3 fraction were purified and re-crystallized. The purified compounds was assumed to be condensed tannins of frame flavan-3-ol frame on the basis of color reagent reaction and to be a mixture of monomer, dimer and trimer according to TLC analysis.

Development and Evaluation of a SYBR Green Real-time PCR Assay for Canine Cytokine Gene Expression (SYBR Green 실시간 역전사 중합효소연쇄반응을 이용한 개 싸이토카인 유전자 발현의 정량)

  • Yu, Do-Hyeon;Ihn, Dong-Chul;Park, Chul;Park, Jin-Ho
    • Journal of Veterinary Clinics
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    • v.27 no.5
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    • pp.508-513
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    • 2010
  • Cytokines are important mediators of the immune response, and quantitating cytokine mRNA is a highly sensitive and attractive method for measuring cytokine production. The objective of the current study was to develop and validate a SYBR green quantitative real-time reverse transcriptase PCR (qRT-PCR) assay for measuring canine cytokine mRNA. The optimal annealing temperatures ($T_a$) of the designed primers were $62^{\circ}C$ for interleukin (IL)-$1{\beta}$, IL-6 and IL-10; $60^{\circ}C$ for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and tumor necrosis factor (TNF)-${\alpha}$; and $58^{\circ}C$ for high mobility group box 1 (HMGB1). Primer efficiencies of all primers calculated for standard curve samples were between 97.1% and 102.6%. No evidence of secondary structure or primer-dimer formation was seen via melt-curve analysis or gel electrophoresis. The developed qRT-PCR assays are highly specific and sensitive and can be used to quantify gene expression levels of canine cytokines.

Behavior of AKD in AKD-sized Paper by Heating Treatment During Ageing (AKD 사이즈 처리한 종이의 저장중 가열처리에 의한 AKD의 거동)

  • Shin, Young-Doo;Seo, Won-Sung;Cho, Nam-Seok
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.39 no.1 s.119
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    • pp.16-24
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    • 2007
  • This study was performed to investigate the reaction between alkylketene dimer(AKD) and cellulose molecules in AKD-sized paper sheet. AKD was added to highly beaten($80{\pm}3^{\circ}SR$) SwBKP(ca. 0.8% on pulp) in order to have much AKD retention in the paper sheet. This AKD-sized paper sheet was aged at different temperatures, $60^{\circ}C,\;80^{\circ}C,\;105^{\circ}C\;and\;125^{\circ}C$. Changes in FT-IR spectra of AKD in paper sheet during the ageing were measured. In addition, sizing degrees of the AKD-sized paper sheet pretreated for 30 sec. at $105^{\circ}C$ were measured by HST size tester during the storage at different temperature. IR spectra of AKD-sized paper sheet preheated at $105^{\circ}C$ for 30 sec. showed unchanged spectra two absorption bands at $1849cm^{-1}\;and\;1722cm^{-1}$ which refer to the typical AKD IR bands. However, these typical AKD bands were gradually reduced with increasing ageing, completely disappeared after 6 hrs. and formed new absorption band at $1706cm^{-1}$, which refers to carbonyl stretching vibration of dialkylktone. Eventually the AKD molecule was hydrolyzed to diakylketone without formation of ${\beta}$-ketoester with cellulose in paper sheet. After 6 days ageing, a little amount of ${\beta}$-ketoester bands was identified in 6 or 7 days ageing, because of the absence of water due to long-term heating. The same tendency was observed at different ageing conditions. At the practical papermaking process, AKD reacts prevailing with water, and mostly seems to be hydrolyzed to dialkylketene. Concerned to the sizing development, AKD-sized paper sheet was shown no sizing development at the initial stage of ageing at $60^{\circ}C$ after heating treatment at $105^{\circ}C$ for 30 sec., and gradually increased the sizing degree with increasing ageing, such as Hercules Sizing Tester (HST) 130 see for 12 hr, HST 300 sec. for 3 days and HST 400 sec. for 5 days. It was concluded that hydrolyzed AKD could contributed to the sizing development of the paper sheet.

The Effect of Hyulbuchuko-tang on a Case with Deep Vein Thrombosis (DVT) and Intracranial Hemorrhage (ICH) (뇌출혈로 항응고제 사용이 불가능한 심부 정맥혈전증에 대한 혈부축어탕 치험례)

  • Kang, Ji-Suck;Park, Sung-Hwan;Song, Moon-Koo;Ahn, Young-Min;Ahn, Se-Young;Lee, Byung-Cheol
    • The Journal of Internal Korean Medicine
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    • v.30 no.2
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    • pp.438-449
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    • 2009
  • Objectives: Deep vein thrombosis (DVT) is a common complication among stroke patients. The implication of DVT progressing into a fatal pulmonary embolism is one of the main reasons treatment cannot be delayed. However, when there is a contradiction for anticoagulants, such intracranial hemorrhage (ICH), it is difficult to determine the course of treatment. Our team reports a case with both acute DVT and ICH who improved with herbal medicine Hyulbuchuko-tang. Methods : A patient with a variety of thrombosis risk factors (atrial fibrillation, DVT, Cb-inf with intracranial hemorrhage due to thrombolytic complications) showed classic symptoms of DVT (pain, edema, discoloration), disorientation and chest discomfort. The patient was administered Hyulbuchuko-tang three times a day for 24 days without any anticoagulants. Conservative therapy including elastic stocking and leg elevation was co-administered. Laboratory tests and extremity vascular Doppler sonography were carried out 3 times during the treatment period. Results : After our treatment period, both popliteal vein DVT and calf vein DVT were not discovered by sonography, and thrombosis derived factors (eg. D-dimer, fibrinogen) decreased. There was no sign of edema or discoloration after treatment, and the patient no longer complained of leg pain, disorientation or chest discomfort. Conclusion : From these results, we suggest that there is a positive effect of Hyulbuchuko-tang on DVT. Hyulbuchuko-tang should be considered as a treatment option when western medical procedures are unavailable.

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The outcome of short-term low-dose aspirin treatment in Kawasaki disease based on inflammatory markers

  • Yoo, Jae Won;Kim, Ji Mok;Kil, Hong Ryang
    • Clinical and Experimental Pediatrics
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    • v.60 no.1
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    • pp.24-29
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    • 2017
  • Purpose: Previously, Kawasaki disease (KD) treatment with low-dose aspirin was administered for 6-8 weeks after the acute phase. However, inflammatory marker levels normalize before 6-8 weeks. In this study, we aimed to investigate the clinical outcome of short-term low-dose aspirin treatment based on inflammatory and thrombotic marker levels. Methods: We performed a retrospective review of the medical records of patients with KD who were hospitalized at Chungnam National University Hospital between September 2012 and May 2014. When fever subsided, low-dose aspirin treatment was started. Inflammatory (white blood cell count, erythrocyte sedimentation rate, and C-reactive protein) and thrombotic markers (D-dimer) were monitored at follow-ups conducted in 1- to 2-week intervals. The low-dose aspirin administration was terminated when both markers were normalized and no cardiovascular complications were observed. Results: Eighty-four patients with KD (complete KD, n=49; incomplete KD, n=35) were enrolled. The inflammatory and thrombotic marker levels were normalized within 3-4 weeks on average. At the beginning the low-dose aspirin treatment, 9 patients had coronary artery lesions but 75 did not. When the low-dose aspirin administration was terminated at the time the inflammatory marker levels were normalized, no new CALs developed during the follow-up at 6-8 weeks. Conclusion: Most of the inflammatory marker levels were normalized within 3-4 weeks after the acute phase of KD. New cardiovascular complications did not develop during the course of the short-term aspirin treatment based on the inflammatory marker levels, clinical findings, and echocardiography.

Reduced Regional Cerebral Blood Flow in Patients with Traumatic Brain Injury Who Had No Structural Abnormalities on Magnetic Resonance Imaging : A Quantitative Evaluation of Tc-99m-ECD SPECT Findings (정상 MRI 소견을 보이는 외상성 뇌손상 환자에서 국소뇌혈류량의 이상)

  • Kim, Nam-Hee;Chung, Young-Ki
    • Korean Journal of Biological Psychiatry
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    • v.9 no.2
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    • pp.152-158
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    • 2002
  • Background & Purpose:Neuropsychological disorders after traumatic brain injury(TBI) are poorly correlated with structural lesions detected by structural neuroimaging techniques such as computed tomography(CT) scan or magnetic resonance imaging(MRI). It is well known that patients with TBI have cognitive and behavioral disorders even in the absence of structural lesions of the brain. This study investigated whether there are abnormalities of regional cerebral blood flow(rCBF) in TBI patients without structural abnormality on MRI, using technetium 99m ethyl cysteinate dimer(Tc-99m-ECD) single photon emission computed tomography(SPECT) scans. Materials and Methods:Twenty-eight TBI patients without structural abnormality on MRI(mild, n=13/moderate, n=9/severe, n=6) and fifteen normal controls were scanned by SPECT. A voxel-based analysis using statistical parametric mapping(SPM) was performed to compare the patients with the normal controls. Results:rCBF was reduced in the right uncus and the right lateral orbitofrontal gyrus in the TBI patients. However, no increase of rCBF was noted in the patients in comparison to the normal controls. Conclusions:These results suggest that the TBI patients, even in the absence of structural lesion of the brain, may have dysfunction of the brain, particularly of the orbitofrontal and anterior pole of the temporal cortex. They also suggest that SPECT can be a useful method to identify brain dysfunctions in combination with structural brain imaging and neuropsychological tests.

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