• Title/Summary/Keyword: high-level synthesis

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Carnitine and Calmodulin N-Methylation in Rat Testis; Calmodulin May beInvolved in Carnitine Biosynthesis

  • Oh, Suk-Heung;Cha, Youn-Soo;Sohn, Hee-Sook
    • Preventive Nutrition and Food Science
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    • v.3 no.3
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    • pp.251-255
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    • 1998
  • Rat testis known to contain all of the enzymes required for carnitine biosynthesis also contains high concentration of calmodulin, a protein which may or may not contain trimethyllysine, the major substrate in carnitine biosynthesis. The purpose of this study was to investigate the levels of carnitine and the state of calmodulin N-methylation in rat testes, and to discuss the possibility of the involvement of calmodulin incarnitine biosynthesis. Nonesterified carnitine , acid soluble acyl carnitine, and acid insoluble acyl carnitine of ra tests were 273 nmole, 62nmole, and 4 nmole/g tissue, respectively. Total carnitine level was 339 nmole/g testes tissue. Calmodulin purified from rat tests was assayed for methylation potential using N-methyltransferase from the rat testes. Rat testes calmodulin showed no 3H-methyl incorporation indicating that the calmodulin was trimethylated already by endogenous calmodulin N-methyltransferase. Amino acid composition analysis revealed that the rat testes calmodulin containd one mole of trimethyllysine per mole of calmodulin. These data suggest that testes calmodulin could provide the trimethyllysine needed for the synthesis of carnitine in the rat tests.

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Free Radical Scavenging Activity and Inhibition of Linoleic Acid Peroxidation of Commercial Tocotrienol Fraction

  • Kim, Joo-Shin;Chung, Hau-Yin
    • Preventive Nutrition and Food Science
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    • v.12 no.3
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    • pp.177-180
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    • 2007
  • Tocotrienols (T3) are minor plant constituents found abundantly in rice bran, which provide a significant source of vitamin E in animal feeds. T3 was reported to have an intrinsic hypocholesterolemic effect by inhibiting HMG-Co A reductase. It has similar antioxidative properties as tocopherols in food and biological system due to their similar chemical structures. However, the antioxidant activity and mechanism of T3 to scavenge free radicals and to inhibit the peroxidation of linoleic acid are less understood. The purpose of this study was to investigate the scavenging effect of T3 on free radicals and its inhibition of peroxide formation. Free radical scavenging activity was monitored by the DPPH (1,1-diphenyl-2-picrylhydrazyl) method whereas inhibition of linoleic acid peroxidation was evaluated using the thiocyanate method. Thiobarbituric acid (TBA) test was used to determine malonaldehyde formation from linoleic acid peroxidation. Free radical scavenging activity increased with increasing concentration levels of T3. T3 exhibited 38.2, 78.6, 92.7 and 96.2% radical scavenging activity at concentrations of 2, 8, 32 and 128 ppm, respectively. At 128 ppm, it was highly effective in inhibiting linoleic acid peroxidation. The activity of T3 evaluated by the thiocyanate method showed low absorbance values indicating a high level of antioxidant activity. All treatments showed similar trends in antioxidant activity when evaluated by both the thiocyanate method and TBA test.

Subcellular Location of Spodpotera Cell-expressed Human HepG2-type Glucose Transport Protein

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.160-164
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    • 2012
  • The baculovirus/insect cell expression system is of great value for the large-scale production of normal and mutant mammalian passive glucose-transport proteins heterologously for structural and functional studies. In most mammalian cells that express HepG2, this transporter isoform is predominantly located at the cell surface. However, it had been reported that heterologous expression of other membrane proteins using the baculovirus system induced highly vacuolated cytoplasmic membranes. Therefore, how a cell responds to the synthesis of large amounts of a glycoprotein could be an interesting area for investigation. In order to examine the subcellular location of the human HepG2 transport proteins when expressed in insect cells, immunofluorescence studies were carried out. Insect cells were infected with the recombinant baculovirus AcNPVHIS-GT or with wild-type virus at a MOI of 5, or were not exposed to viral infection. A high level of fluorescence displayed in cells infected with the recombinant virus indicated that transporters are expressed abundantly and present on the surface of infected Sf21 cells. The evidence for the specificity of the immunostaining was strengthened by the negative results shown in the negative controls. Distribution of the transporter protein expressed in insect cells was further revealed by making a series of optical sections through an AcNPVHIS-GT-infected cell using a confocal microscope, which permits optical sectioning of cell sample. These sections displayed intense cytoplasmic immunofluorecence surrounding the region occupied by the enlarged nucleus, indicating that the expressed protein was present not only at the cell surface but also throughout the cytoplasmic membranous structures.

Present Status and Future Prospect of Quantum Dot Technology (양자점 (Quantum dot) 기술의 현재와 미래)

  • Hong, H.S.;Park, K.S.;Lee, C.G.;Kim, B.S.;Kang, L.S.;Jin, Y.H.
    • Journal of Powder Materials
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    • v.19 no.6
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    • pp.451-457
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    • 2012
  • Nowadays, research and development on quantum dot have been intensively and comprehensively pursued worldwide in proportion to concurrent breakthrough in the field of nanotechnology. At present, quantum dot technology forms the main interdisciplinary basis of energy, biological and photoelectric devices. More specifically, quantum dot semiconductor is quite noteworthy for its sub-micro size and possibility of photonic frequency modulation capability by controlling its size, which has not been possible with conventionally fabricated bulk or thin film devices. This could lead to realization of novel high performance devices. To further understand related background knowledge of semiconductor quantum dot at somewhat extensive level, a review paper is presently drafted to introduce basics of (semiconductor) quantum dot, its properties, applications, and present and future market trend and prospect.

INTRACELLULAR AMINO ACID PROFILE OF RUMEN BACTERIA AS INFLUENCED BY UREA FEEDING AND ITS DURATION

  • Kobayashi, Y.;Wakita, M.;Hoshino, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.619-622
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    • 1993
  • Rumen bacterial amino acids in sheep on urea diet were monitored to assess a possible change in amino acid synthesis as a long term response to high rumen ammonia environment. A sheep was fed a semipurified diet with soybean meal, followed by a diet with urea as a main nitrogen source. Mixed rumen bacteria were harvested from ruminal fluid taken 3 h after feeding (twice in soybean meal feeding and 6 times in urea feeding) and fractionated as cell wall, proteins and protein-free cell supernatant of monitor amino acids in each fraction. Ruminal ammonia concentration at the sampling ranged from 5.7 to 39.5 mgN/dl. Cell wall and protein fractions of mixed rumen bacteria were stable in their amino acid composition regardless of nitrogen sources of diet and the feeding duration. However, protein-free cell supernatant fraction showed a higher alanine proportion with urea feeding (18.6 and 28.2 molar % of alanine for samples from sheep fed soybean meal and urea, respectively) and its duration (20.6 and 32.9 molar % for samples from sheep on urea diet for 1 and 65 days, respectively). Total free amino acid level of bacteria was depressed in the initial period of urea feeding but restored on 65th day of the feeding. These results suggest that an alanine synthesizing system may develop in rumen bacteria as urea feeding becomes longer.

MR304-1, A Melanin Synthesis Inhibitor Produced by Trichoderma harzianum (Trichoderma harzianum이 생산하는 melanin 생성 저해물질 MR304-1)

  • Lee, Choong-Hwan;Chung, Myung-Chul;Lee, Ho-Jae;Lee, Ke-Ho;Kho, Yung-Hee
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.641-646
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    • 1995
  • During the screening of inhibitors of melanin biosynthesis from microbial secondary metabolites, a fungal strain MR304 which was capable of producing high level of an inhibitor was selected. Based on taxonomic studies, this fungus could be classified as Trichoderma harzianum. The active compound (MR304-1) was purified from culture broth by Diaion HP-20 column chromatography, ethylacetate extraction, Sephadex LH-20 column chromatographv and HPLC. The inhibitor was identified as 3-(1,5-dihvdroxy-3-isocyanocyclopent-(E)-3-envl)prop-2-enoate by spectroscopic methods of UV, ESIMS, $^{1}$H-NMR, $^{13}$C-NMR, NOE, HMQC and HMBC. MR304-1 showed strong mushroom tyrosinase inhibitory activity with IC$_{50}$ value of 0.25 $\mu $g/ml. It inhibited melanin biosynthesis with 15 mm inhibition zone at 30 $\mu $g/paper disc in Streptomyces bikiniensis, a bacterium used as an indicator organism in this work. It also inhibited melanin biosynthesis in B16 melanoma cells with a niinimum inhibitory concentration of 0.05 $\mu $g/ml.

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Di-acetyl-nor-aporphines: Novel molecules and a novel mechanism to inhibit melanogenesis

  • Lintner, Karl;Peschard, Olivier;Leroux, Richard;Mondon, Philippe;Lamy, Francois
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.268-284
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    • 2003
  • Nor-aporphine derivatives have been discovered which interfere with the flux of Calcium into and out of the cell interior. It has been shown that adrenergic antagonists that block the Calcium exchange lead to an inhibition of Protein kinase C activity, thus blocking tyrosinase activation. Di-acetyl-dimethoxy-methyl-nor-aporphine is a semi-synthetic molecule of natural origin with very high potency. On B16 melanocytes as well as in normal human melanocytes the decrease in melanin synthesis reaches -50% at a level of 40 ppm in the culture medium. On a molar concentration basis, this is 50 to 70 times stronger than Kojic acid inhibition. Yet, the cell viability is not affected. Reversibility studies show that after washing out of the active compound, melanogenesis returns to normal levels. Possible mechanisms of the activity are discussed. Tests carried out on SkinEthic(R) three-dimensional models of the epidermis and in vivo clinical studies on Asian population confirm the strong inhibition of melanogenesis. Safety evaluation of these molecules, on the other hand, demonstrates good skin tolerance and absence of toxicity.

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Synthesis and Characterization of Low Molecular Weight Poly(methyl acrylate)-b-Polystyrene by a Combination of ATRP and Click Coupling Method

  • Hasneen, Aleya;Kim, Su-Jeong;Paik, Hyun-Jong
    • Macromolecular Research
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    • v.15 no.6
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    • pp.541-546
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    • 2007
  • The combination of atom transfer radical polymerization (ATRP) and click chemistry was employed for the efficient preparation of well-defined block copolymers. Bromo terminated poly(methyl acrylate) (pMA-Br) was prepared by an ATRP initiator, ethyl-2-bromoisobutyrate (EBiB). Subsequently, the bromine chain end of pMA-Br was converted to an azide group by simple nucleophilic substitution reaction. ${\alpha}-Alkyn-{\omega}-bromo-functionalized$ polystyrene was also synthesized by ATRP using the alkyn-functionalized initiator, propargyl-2-bromoisobutyrate (PgBiB). In both cases, the conversion was limited to a low level to ensure a high degree of chain end functionality. Then the coupling reaction between the azide end group in $pMA-N_3$ and alkyn-functionalized PgBiB-pSt was performed by Cu(I)catalysis. This coupling reaction was monitored by gel permeation chromatography (GPC). The synthesized block copolymer was characterized by FT-IR, $^1H-NMR$ spectroscopy and $^1H-^1H$ COSY correlation spectroscopy.

A Study on Constructing the System-on-Chip based on Embedded Systems (임베디드시스템에 기반을 둔 시스템온칩 구성에 관한 연구)

  • Park, Chun-Myoung
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2015.05a
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    • pp.888-889
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    • 2015
  • This paper presents a method of constructing the system-on-chip(SoC) based on embedded systems. The proposed method is more compact and effectiveness than former methods. The requirements generation start high level performance simulation and then passes to an executable specification suitable for implementation using a hardware/software co-design tool. The reuse of pre-exiting components is supported, as well as synthesis of the system interface, but only after much work is done to program the hardware/software co-design tool. The actual design flow described allows feedback among all design levels, e.g. from implementation up to requirements, throughout the process. In the future, it is necessary to development the advanced method of constructing system-on-chip based on embedded systems.

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A Rapid and Simple Method for Construction and Expression of a Synthetic Human Growth Hormone Gene in Escherichia coli

  • Roytrakul, Sittiruk;Eurwilaichitr, Lily;Suprasongsin, Chittiwat;Panyim, Sakol
    • BMB Reports
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    • v.34 no.6
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    • pp.502-508
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    • 2001
  • A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

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