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L-DOPA Synthesis Using Tyrosinase-immobilized on Electrode Surfaces

  • Rahman, Siti Fauziyah;Gobikhrisnan, Siramulu;Gozan, Misri;Jong, Gwi Taek;Park, Don-Hee
    • Korean Chemical Engineering Research
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    • v.54 no.6
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    • pp.817-821
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    • 2016
  • Levodopa or L-3,4-dihydroxyphenylalanine (L-DOPA) is the direct precursor of the neurotransmitter dopamine. L-DOPA is a well-known neuroprotective agent for the treatment of Parkinson's disease symptoms. L-DOPA was synthesized using the enzyme, tyrosinase, as a biocatalyst for the conversion of L-tyrosine to L-DOPA and an electrochemical method for reducing L-DOPAquinone, the product resulting from enzymatic synthesis, to L-DOPA. In this study, three electrode systems were used: A glassy carbon electrode (GCE) as working electrode, a platinum, and a Ag/AgCl electrode as auxiliary and reference electrodes, respectively. GCE has been modified using electropolymerization of pyrrole to facilitate the electron transfer process and immobilize tyrosinase. Optimum conditions for the electropolymerization modified electrode were a temperature of $30^{\circ}C$ and a pH of 7 producing L-DOPA concentration 0.315 mM. After 40 days, the relative activity of an enzyme for electropolymerization remained 38.6%, respectively.

Acrolein, the toxic endogenous aldehyde, induces neurofilament-L aggregation

  • Jeong, Moon-Sik;Kang, Jung-Hoon
    • BMB Reports
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    • v.41 no.9
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    • pp.635-639
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    • 2008
  • Acrolein is a highly reactive by product of lipid peroxidation and individuals with neurodegenerative disorders have been shown to contain elevated concentrations of this molecule in the brain. In the present study, we examined the pattern of neurofilament-L (NF-L) modification elicited by acrolein. When NF-L was incubated with acrolein, protein aggregation occurred in a acrolein concentration-dependent manner. Exposure of NF-L to acrolein also led to the generation of protein carbonyl compounds. Through the addition of free radical scavengers we observed a significant decrease in acrolein-mediated NF-L aggregation. These results indicate that free radicals may be involved in the modification of NF-L by acrolein. In addition, dityrosine crosslink formation was observed in acrolein-mediated NF-L aggregates and these aggregates displayed thioflavin T reactivity, reminiscent of amyloid. This study suggests that acrolein-mediated NF-L aggregation might be closely related to oxidative reactions, thus these reactions may play a critical role in neuro-degenerative diseases.

Conversion of D-$\alpha$-Amino-$\varepsilon$-Caprolactam into L-Lysine Using Cell-free Extracts of Alcaligenes eutrophus A52 (Alcaligenes eutrophus A52의 무세포 추출액에 의한 D-$\alpha$-Amino-$\varepsilon$-Caprolactam으로부터 L-Lysine으로의 전환)

  • 박희동;최선택;이인구
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.375-380
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    • 1987
  • D-$\alpha$-Amino-$\varepsilon$-carpolactam racemase (EC 5.1.1) and L-$\alpha$-amino-$\varepsilon$-caprolactam hydrolase (EC 3.5.2) were fractionated from cell-free extracts of Alcaligenes eutrophus A52 using ammonium sulfate precipitation and DEAE-cellulose ion exchange chromatography. It was made sure that D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-$\alpha$-amino-$\varepsilon$-caprolactam by racemase, and then hydrolyzed into L-lysine by hydrolase in Alcaligenes eutrophus A52. For the conversion of D-$\alpha$-amino-$\varepsilon$-caprolactam into L-lysine by cell-free extracts of Alcaligenes eutrophus A52, the optimum temperature and pH were 6$0^{\circ}C$ and 8.5 respectively. The results showed that 0.5% D-$\alpha$-amino-$\varepsilon$-caprolactam was converted to L-lysine at 55$^{\circ}C$ for 10 hr with a conversion rate of 98% by cell-free extracts containing 3.1mg of protein.

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Heterologous Expression of Human $\beta$-Defensin-1 in Bacteriocin-Producing Laetoeoeeus lactis

  • CHOI HAK JONG;SEO MYUNG JI;LEE JUNG CHOUL;CHEIGH CHAN ICK;PARK HOON;AHN CHEOL;PYUN YU RYANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.330-336
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    • 2005
  • Lactococcus lactis A164 is a nisin Z-producing strain isolated from kimchi. Its antimicrobial spectrum has been found to be active against most Gram-positive bacteria tested, yet inactive against Gram-negative bacteria [3]. Accordingly, to overcome this drawback, the current study attempted to express human $\beta$-defensin-l (hBD-l), which kills both Gram-positive and Gram-negative bacteria in L. lactis AI64. When the hBD-l cDNA was introduced using a nisin Z-controlled expression cassette, the L. lactis A164 transformants grew very poorly, due to the bactericidal effect of the expressed hBD-l against the transformants. Therefore, a gene fusion system was designed to reduce the toxicity of the expressed heterologous protein against the host cells. As such, the hBD-l gene was fused to the DsbC- Tag of pET -40b(+), then introduced to L. lactis A 164. The transformants expressed an intracellular 35.6-kDa DsbC-hBD-l fusion protein that exhibited slight activity against the host cells, yet not enough to strongly inhibit the cell growth. To obtain the recombinant hBD-l, the DsbC-hBD-l fusion protein was purified by nickel-affinity column chromatography, and the DsbC-Tag removed by cleaving with enterokinase. The cleaved mature hBD-l exhibited strong bactericidal activity against E. coli JM109, indicating that the recombinant L. lactis A 164 produced a biologically active hBD-I. In addition, the recombinant L. lactis A 164 was also found to produce the same level of nisin Z as the wild-type.

On the Representations of Finite Distributive Lattices

  • Siggers, Mark
    • Kyungpook Mathematical Journal
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    • v.60 no.1
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    • pp.1-20
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    • 2020
  • A simple but elegant result of Rival states that every sublattice L of a finite distributive lattice 𝒫 can be constructed from 𝒫 by removing a particular family 𝒥L of its irreducible intervals. Applying this in the case that 𝒫 is a product of a finite set 𝒞 of chains, we get a one-to-one correspondence L ↦ 𝒟𝒫(L) between the sublattices of 𝒫 and the preorders spanned by a canonical sublattice 𝒞 of 𝒫. We then show that L is a tight sublattice of the product of chains 𝒫 if and only if 𝒟𝒫(L) is asymmetric. This yields a one-to-one correspondence between the tight sublattices of 𝒫 and the posets spanned by its poset J(𝒫) of non-zero join-irreducible elements. With this we recover and extend, among other classical results, the correspondence derived from results of Birkhoff and Dilworth, between the tight embeddings of a finite distributive lattice L into products of chains, and the chain decompositions of its poset J(L) of non-zero join-irreducible elements.

Effects of Diospyros kaki L. Radix or Diospyros kaki L. Folium on Arterial Contraction Induced by Phenylephrine in Rabbit (시근과 시엽이 Phenylephrine으로 유발된 가토의 수축혈관에 미치는 영향)

  • 김희철;남창규;김호현;성현제
    • The Journal of Korean Medicine
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    • v.24 no.1
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    • pp.141-154
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    • 2003
  • Objectives : This study was undertaken to define the effect of Diospyros kaki L. Radix or Diospyros kaki L. Folium on phenylephrine-induced arterial contraction and the mechanism of Diospyros kaki L. Radix or Diospyros kaki L. Foliuminduced relaxation. Methods : In order to investigate the effect of Diospyros kaki L. Radix or Diospyros kaki L. Folium on contracted rabbit carotid arterial strips, transverse strips with intact or damaged endothelium were used for the experiment using organ bath. Diospyros kaki L. Radix or Diospyros kaki L. Folium extract was infused into contracted arterial strips induced by phenylephrine. To analyze the mechanism of Diospyros kaki L. Radix or Diospyros kaki L. Folium-induced relaxation, Diospyros kaki L. Radix or Diospyros kaki L. Folium extract was infused into contracted arterial strips induced by phenylephrine after treatment with indomethacin, $N{\omega}-nitro-L-arginine$, methylene blue or tetraethylammonium chloride, and $Ca^{2+}$ was infused into contracted arterial strips induced by phenylephrine after treatment of Diospyros kaki L. Radix or Diospyros kaki L. Folium in a $Ca^{2+}$-free solution. Results : Diospyros kaki L. Radix or Diospyros kaki L. Folium showed relaxation effect on arterial strip with endothelium contracted by phenylephrine, but in the strips without endothelium, Diospyros kaki L. Radix or Diospyros kaki L. Folium-induced relaxation was significantly inhibited. The endothelium-dependent relaxation induced by Diospyros kaki L. Radix or Diospyros kaki L. Folium was decreased by pretreatment with $N{\omega}-nitro-L-arginine$ or methylene blue but it was not observed in the strips pretreated with indomethacin or tetraethylammonium chloride. When $Ca^{2+}$ was applied to the strips which were contracted by phenylephrine in a $Ca^{2+}$-free solution, arterial contraction was increased. However, pretreatment with Diospyros kaki L. Radix or Diospyros kaki L. Folium inhibited contractile response to $Ca^{2+}$. Conclusions : Diospyros kaki L. Radix or Diospyros kaki L. Folium may suppress influx of extra- cellular $Ca^{2+}$ through the formation of nitric oxide in the vascular endothelial cells.

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Study on Myocardial Protective Effect of L-arginine (L-Arginine의 흰쥐 적출심근보호 효과에 관한 연구)

  • 이인성;이헌재
    • Journal of Chest Surgery
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    • v.29 no.10
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    • pp.1076-1080
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    • 1996
  • The results about the myocardial protection of recta of the nitric oxide precursor L-arginine upon reperrusion injury after ischemia are diverse. These diversities may be model dependent. Experiments were designed and performed to investigate myocardial protection effects according to the concentration of L-arginine. The Isolated rat hearts were subjected in a 30 minutes oi normothermic ischemia and reperfused for 30 minutes with reperfusate containing 0, 1, 2, 3, 4 moil L-arginine. After 30 minutes of reperfusion, group with 1 and 2 mM/L L-arginine showed a trend of better recovery in left ventricular systolic function(left ventricular developed pressure, positive maximum dpfdt), diastolic function(negative maximum dpfdt) and coronary flow compared to control group(reperfusate no L-arginine). Recovery was impaired with a higher concentration, and at 4 moil L-arginine r covery was worse than control(p (0, 05). These results suggest that optimal concentration of L-arginine Is Important or the recovery of myocardial and endothelial function after ischemia and reperfusion.

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A Study on Growth-inhibiting Protein of Human Cancer Cells Secreted from 373-L1 Cell-line (3T3-L1 세포주해서 분비하는 인체 암세포 성장억제 단백질에 대한 연구)

  • Eun, Jae-Soon;Kweon, Jin
    • Biomolecules & Therapeutics
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    • v.4 no.1
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    • pp.46-50
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    • 1996
  • Inhibition of the growth of human cancer cells by proteins secreted from 373-L1 cells was investigated in the present study. The growth of human cancer cells was inhibited by co-culture with 373-L1 cells under 10% FBS and DME, DME, GIT and serumless medium, respectively. The conditioned medium of cultured 373-L1 cells under serumless medium was concentrated 100-fold through an ultrafiltration cell with a 10,000 molecular weight cutoff at 4$^{\circ}C$ under positive pressure using nitrogen(373-L1 EM). 373-L1 EM inhibited the growth of HeLa, Hep G 2, KHOS-Np, A43l and MCF-7 cells. 3T3-L1 EM was purified with FPLC, DEAE-ion exchange chromatography and phenyl-sepharose chromatography. The major protein of 373-L1 EM has a molecular weight of 66,000-68,000 in SDS-PAGE analysis. The results suggest that the inhibitory activity of 373-L1 EM appears to be due to some protein(m.w.66,000-68,000) secreted by 373-L1 cells.

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Effects of Calcium Lactate and Chungkukjang on Calcium Status in Rat

  • Lee, Ye-Kyung;Lee, Myung-Ye;Kim, Mee-Kyung;Choe, Won-Kyung;Kim, Soon-Dong
    • Preventive Nutrition and Food Science
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    • v.9 no.1
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    • pp.45-52
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    • 2004
  • Effects of dietary calcium lactate (CaL-A) and Chungkukjang (Korean native fermented soybean) on bone mass, calcium status, body weight, serum glucose and cholesterol levels in young male rats were investigated. Chungkukjang was fermented by mixing 4 types of Bacillus sp., and then dried at 45$^{\circ}C$. Calcium lactate was prepared from the ash of black snail. The rats were fed a commercial rat diet for 1 week and then the experimental diets for 4 weeks. Animals were divided into four dietary groups: one calcium-deficient diet (Ca-De) and one of three with calcium supplemented diets (5 g calcium/kg diet) with either calcium phosphate (Ca-P), CaL-A, or CaL-A + Chuntkukjang (CaL-AC). Calcium supplemented diets contained 39 g Ca-P/kg diet and 28 g/kg of calcium lactate in the CaL-A and CaL-AC diets. Body weight gains during the 4 weeks in the Ca-P, CaL-A, CaL-AC and Ca-De groups were 130.45 g,112.50 g, 143.40 g and 10.20 g, respectively. Feed consumption of the groups from high to low was CaL-AC > Ca-P > CaL-A > Ca-De. The Ca-De group had low femur weights and low serum calcium concentrations, while they were comparatively high in CaL-AC, Ca-P and CaL-A groups. The Ca-De groups excreted less calcium in urine than did the other rats, probably due to increased absorption of the mineral in Ca-P, CaL-A and CaL-AC groups. Microscopic observations revealed that there were many regularly spaced holes in the femur of Ca-De group, while there were much smaller regularly spaced holes in Ca-P group. However, no holes in femur were observed in the CaL-A and CaL-AC groups. Bone surfaces were especially smooth and clean in the CaL-AC group. Serum concentrations of glucose and total cholesterol were remarkably lower in the CaL-AC group than in the other supplemented groups. These results suggest that calcium from CaL-A has higher bioavailability than from Ca-P, and dietary Chungkukjang may have a beneficial effect on calcium metabolism.

Couette-Poiseuille flow based non-linear flow over a square cylinder near plane wall

  • Bhatt, Rajesh;Maiti, Dilip K.;Alam, Md. Mahbub;Rehman, S.
    • Wind and Structures
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    • v.26 no.5
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    • pp.331-341
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    • 2018
  • A numerical study on the flow over a square cylinder in the vicinity of a wall is conducted for different Couette-Poiseuille-based non-uniform flow with the non-dimensional pressure gradient P varying from 0 to 5. The non-dimensional gap ratio L (=$H^{\ast}/a^{\ast}$) is changed from 0.1 to 2, where $H^{\ast}$ is gap height between the cylinder and wall, and $a^{\ast}$ is the cylinder width. The governing equations are solved numerically through finite volume method based on SIMPLE algorithm on a staggered grid system. Both P and L have a substantial influence on the flow structure, time-mean drag coefficient ${\bar{C}}_D$, fluctuating (rms) lift coefficient ($C_L{^{\prime}}$), and Strouhal number St. The changes in P and L leads to four distinct flow regimes (I, II, III and IV). Following the flow structure change, the ${\bar{C}}_D$, $C_L{^{\prime}}$, and St all vary greatly with the change in L and/or P. The ${\bar{C}}_D$ and $C_L{^{\prime}}$ both grow with increasing P and/or L. The St increases with P for a given L, being less sensitive to L for a smaller P (< 2) and more sensitive to L for a larger P (> 2). A strong relationship is observed between the flow regimes and the values of ${\bar{C}}_D$, $C_L{^{\prime}}$ and St. An increase in P affects the pressure distribution more on the top surface than on bottom surface while an increase in L does the opposite.