• Title/Summary/Keyword: $N_f$ Line

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D-Methionine and 2-hydroxy-4-methylthiobutanoic acid i alter beta-casein, proteins and metabolites linked in milk protein synthesis in bovine mammary epithelial cells

  • Seung-Woo, Jeon;Jay Ronel V., Conejos;Jae-Sung, Lee;Sang-Hoon, Keum;Hong-Gu, Lee
    • Journal of Animal Science and Technology
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    • v.64 no.3
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    • pp.481-499
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    • 2022
  • This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/ F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.

Soft Plasma Flash X-ray Generator Utilizing a Vacuum Discharge Capillary

  • Sato, Eiichi;Hayasi, Yasuomi;Usuki, Tatsumi;Sato, Koetsu;Takayama, Kazuyoshi;Ido, Hideaki
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2002.09a
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    • pp.400-403
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    • 2002
  • The fundamental experiments for measuring soft x-ray characteristics from the vacuum capillary are described. These experiments were primarily performed in order to generate line spectra such as x-ray lasers. The generator consists of a high-voltage power supply, a polarity-inversion ignitron pulse generator, a turbo-molecular pump, and a radiation tube with a capillary. A high-voltage condenser of 200 nF in the pulse generator is charged up to 20 kV by the power supply, and the electric charges in the condenser are discharged to the capillary in the tube after closing the ignitron. During the discharge, weakly ionized plasma forms on the inner and outer sides of a capillary. In the present work, the pump evacuates air from the tube with a pressure of about 1 mPa, and a demountable capillary was developed in order to measure x-ray spectra according to changes in the capillary length. In this capillary, the anode (target) and cathode elements can be changed corresponding to the objectives. The capillary diameter is 2.0 mm, and the length is adjusted from 1 to 50 mm. When a capillary with aluminum anode and cathode electrodes was employed, both the cathode voltage and the discharge current almost displayed damped oscillations. The peak values of the voltage and current increased when the charging voltage was increased, and their maximum values were -10.8 kV and 4.7 kA, respectively. The x-ray durations observed by a 1.6 ${\mu}$m aluminum filter were less than 30 ${\mu}$s, and we detected the aluminum characteristic x-ray intensity using a 6.8 ${\mu}$m aluminum filter. In the spectrum measurement, two sets of aluminum and titanium electrodes were employed, and we observed multi-line spectra. The line photon energies seldom varied according to changes in the condenser charging voltage and to changes in the electrode element. In the case where the titanium electrode was employed, the line number decreased with corresponding decreases in the capillary length. Compared with incoherent visible light, these rays from the capillary were diffracted and diffused greatly after passing through two slits.

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Association of A/T Rich Microsatellites with Responses to Artificial Selection for Larval Developmental Duration in the Silkworm Bombyx mori

  • Pradeep, Appukuttan Nair Retnabhavan;Awasthi, Arvind Kumar;Urs, Raje Siddaraje
    • Molecules and Cells
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    • v.25 no.4
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    • pp.467-478
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    • 2008
  • Simple sequence repeats (SSRs) and interSSR (ISSR) marker systems were used in this study to reveal genetic changes induced by artificial selection for short/long larval duration in the tropical strain Nistari of the silkworm Bombyx mori. Artificial selection separated longer larval duration (LLD) ($29.428{\pm}0.723days$) and shorter larval duration (SLD) ($22.573{\pm}0.839days$) lines from a base, inbred population of Nistari (larval span of $23.143{\pm}0.35days$). SSR polymorphism was observed between the LLD and SLD lines at one microsatellite locus, Bmsat106 ($CA_7$) and at two loci of 1074 bp and 823 bp generated with the ISSR primer UBC873. Each of these loci was present only in the LLD line. The loci segregated in the third generation of selection and were fixed in opposite directions. In the $F_2$ generation of the $LLD{\times}SLD$ lines, the alleles of Bmsat106 and $UBC873_{1074bp}$ segregated in a 1:1 ratio and the loci were present only in the LLD individuals. $UBC873_{823bp}$ was homozygous. Single factor ANOVA showed a significant association between the segregating loci and longer larval duration. Together, the two alleles contributed to an 18% increase in larval duration. The nucleotide sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci had 67% A/T content and consisted of direct, reverse, complementary and palindromic repeats. The repeats appeared to be "nested" (59%) in larger repeats or as clustered elements adjacent to other repeats. Of 203 microsatellites identified, dinucleotides (67.8%) predominated and were rich in A/T and T/A motifs. The sequences of the $UBC873_{1074bp}$ and $UBC873_{823bp}$ loci showed similarity (E = 0.0) to contigs located in Scaffold 010774 and Scaffold 000139, respectively, of the B. mori genome. BLASTN analysis of the $UBC873_{1074bp}$ sequence showed significant homology of (nt.) 45-122 with upstream region of three exons from Bombyx. The complete sequence of this locus showed ~49% nucleotide conservation with transposon 412 of Drosophila melanogaster and the Ikirara insertions of Anopheles gambiae. The A + T richness and lack of coding potential of these small loci, and their absence in the SLD line, reflect the active process of genetic change associated with the switch to short larval duration as an adaptation to the tropics.

Molecular Mechanism of the Antiproliferative Effect by Ginseng Panaxynol on a Human Malignant Melanoma Cell Line, SK-MEL-1 (인체 흑색종 세포주 SK-MEL-1에 대한 인삼 panaxynol의 항증식 효과 기전)

  • Cho Hongkeun;Yu Su-Jin;Roh Joo Young;;Hwang Woo-Ik;Sohn Jeongwon
    • Journal of Ginseng Research
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    • v.23 no.3 s.55
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    • pp.190-197
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    • 1999
  • In this study, the molecular mechanism of the growth inhibitory effect of panaxynol was investigated in a human malignant melanoma cell line, SK-MEL-l. In the cell cycle analysis, panaxynol arrested cell cycle progression of SK-MEL-I at the G1 phase. Immunoblot analysis demonstrated that panaxynol increased $p21^{WAF1}$ and decreased cdc2 expression. Protein levels of pl6, p27, E2F-1, Rb, and p53 were not changed. Thus, the changes in expression levels of $p21^{WAF1}$ and cdc2 apparently mediate the cell cycle arrest caused by panaxynol. In addition, cycloheximide (CHX) partially reversed the growth inhibition by panaxynol, which suggested that new protein synthesis was required. On the other hand, LLnL, a proteasome inhibitor, increased antiproliferative effect of panaxynol. This may be due to stabilization of the protein(s) responsible for the growth inhibition such as $p21^{WAF1}$. In summary, these results demonstrate that panaxynol inhibits proliferation of SK-MEL-I by inducing cell cycle arrest at the G1 phase and the inhibitory effect is mediated by the increased level of $p21^{WAF1}$ as well as decreased cdc2 expression.

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The Effect of Tidal Cycle and River Runoff on the Dynamic of Nutrients in Keum river estuary (금강하구역에서 영양염 거동에 대한 조석 및 담수유출의 영향)

  • Kim, Jong-Gu;Kang, Hoon
    • Journal of Environmental Science International
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    • v.11 no.6
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    • pp.519-528
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    • 2002
  • This study was to evaluate the impact of river runoff and salt intrusion by tide on nutrient balance of estuary during a complete tidal cycle. 24 hours time series survey was carried out during a spring tide July 2001 on a tidal estuary in the Keum river. Three stations(A,B,C) were set along a transect line of about 10km, which linked the lower part of estuary dyke to the subtidal zone. Surface water was sampled simultaneously at each station every hours f3r the determination of nutrients. Water temperature, pH and dissolved oxygen were measured in situ. Riverine input of silicate and nitrate during ebb tide significantly increased the concentration of all stations. Conversely, during high tide, nutrient concentration were lowered by the mixing of fresh water with sea water Ammonium nitrogen concentration were higher at intertidal zone(Stn.B) due to sewage inflow to Kyeongpo stream and ammonium release under anaerobic conditions. Also, these results was discussed as a biological component that influences the processes of nutrient regeneration within the estuary. Best correlations were found at lower part of estuary dyke(Stn.A) for salinity against DIN(Y=0.121 Sal.+4.97, r2=0.956) and silicate(Y=0.040 Sal.+2.62, r2=0.785). But no significant correlation was found between salinity and ammonium. Unbalanced elemental ratio(N/P, Si/N and Si/P) depended significantly on the import of nutrients (silicate & nitrate nitrogen) from river and stream. The effect of the tidal cycle and river runoff is important that in determining the extend of the variations in nutrient concentrations at all station.

Statistical Analysis on Process Variables in Linear Roll-CMP (선형 Roll-CMP에서 공정변수에 관한 통계적 분석)

  • Wang, Han;Lee, Hyunseop;Jeong, Haedo
    • Tribology and Lubricants
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    • v.30 no.3
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    • pp.139-145
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    • 2014
  • Nowadays, most micro-patterns are manufactured during flow line production. However, a conventional rotary chemical mechanical polishing (CMP) system has a limited throughput for the fabrication of large and flexible electronics. To overcome this problem, we propose a novel linear roll-CMP system for the planarization of large-area electronics. In this paper, we present a statistical analysis on the linear roll-CMP process of copper-clad laminate (CCL) to determine the impacts of process parameters on the material removal rate (MRR) and its non-uniformity (NU). In the linear roll-CMP process, process parameters such as the slurry flow rate, roll speed, table feed rate, and down force affect the MRR and NU. To determine the polishing characteristics of roll-CMP, we use Taguchi's orthogonal array L16 (44) for the experimental design and F-values obtained by the analysis of variance (ANOVA). We investigate the signal-to-noise (S/N) ratio to identify the prominent control parameters. The "higher is better" for the MRR and "lower is better" for the NU were selected for obtaining optimum CMP performance characteristics. The experimental and statistical results indicate that the down force and roll speed mainly affect the MRR and the down force and table feed rate determine the NU in the linear roll-CMP process. However, over 186.3 N of down force deteriorates the NU because of the bending of substrate. Roll speed has little relationship to the NU and the table feed rate does not impact on the MRR. This study provides information on the design parameter of roll-CMP machine and process optimization.

Identification and Characterization of a Novel Antioxidant Peptide from Bovine Skim Milk Fermented by Lactococcus lactis SL6

  • Kim, Sang Hoon;Lee, Ji Yoon;Balolong, Marilen P.;Kim, Jin-Eung;Paik, Hyun-Dong;Kang, Dae-Kyung
    • Food Science of Animal Resources
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    • v.37 no.3
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    • pp.402-409
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    • 2017
  • A novel peptide having free radical scavenging activity was separated, using an on-line high-performance liquid chromatography (HPLC) - ABTS screening method, from bovine skim milk fermented by Lactococcus lactis SL6 (KCTC 11865BP). It was further purified using reverse phase-HPLC (RP-HPLC) and sequenced by RP-HPLC-tandem mass spectrometry. The amino acid sequence of the identified peptide was determined to be Phe-Ser-Asp-Ile-Pro-Asn-Pro-Ile-Gly-Ser-Glu-Asn-Ser-Glu-Lys-Thr-Thr-Met-Pro-Leu-Trp (2,362 Da), which is corresponding to the C-terminal fragment of bovine ${\alpha}_{s1}$-casein (f179-199). The hydroxyl radicals scavenging activity ($IC_{50}$ $28.25{\pm}0.96{\mu}M$) of the peptide chemically synthesized based on the MS/MS data showed a slightly lower than that of the natural antioxidant Trolox ($IC_{50}$ $15.37{\pm}0.52{\mu}M$). Furthermore, derivatives of the antioxidant peptide were synthesized. The antioxidative activity of the derivatives whose all three proline residues replaced by alanine significantly decreased, whereas replacement of two proline residues in N-terminal region did not affect its antioxidative activity, indicating that $3^{rd}$ proline in C-terminal region is critical for the antioxidative activity of the peptide identified in this study. In addition, N-terminal region of the antioxidant peptide did not show its activity, whereas C-terminal region maintained antioxidative activity, suggesting that C-terminal region of the peptide is important for antioxidative activity.

A Study of New Technique Development for Creep Evaluation of Heat Resistant Steel Weldment(I) (내열강 용접부의 크리프 평가 신기술 개발에 관한 연구)

  • 유효선;백승세;권일현;이송인
    • Journal of Welding and Joining
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    • v.20 no.6
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    • pp.30-30
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    • 2002
  • It has been reported that the creep characteristics on weldment which is composed of weld metal(W.M), fusion line(F.L), heat-affected zone(HAZ), and base meta(B.M) could be unpredictably changed in severe service conditions such as high temperature and high pressure. However, the studies done on creep damage in power plant components have been mostly conducted on B.M and not the creep properties of the localized microstructures in weldment have been thoroughly investigated yet. In this paper, it is investigated the creep characteristics for three microstructures like coarse-grain HAZ(CGHAZ), W.M, and B.M in X20CrMoV121 steel weldment by the small punch-creep-(SP-Creep) test using miniaturized specimen(l0×10×0.5mm). The W.M microstructure possesses the higher creep resistance and shows lower creep strain rate than the B.M and CGHAZ. In the lower creep load the highest creep strain rate is exhibited in CGHAZ, whereas in the higher creep load the B.M represents the high creep strain rate. The power law correlation for all microstructures exists between creep rate and creep load at 600℃. The values of creep load index (n) based on creep strain rate for B.M, CGHAZ, and W.M are 7.54, 4.23, and 5.06, respectively and CGHAZ which shows coarse grains owing to high welding heat has the lowest creep loade index. In all creep loads, the creep life for W.M shows the highest value.

CMnAl TRIP Steel Surface Modification During CGL Processing

  • Gong, Y.F.;Lee, Y.R.;Kim,, Han-S.;Cooman, B.C.De
    • Corrosion Science and Technology
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    • v.9 no.2
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    • pp.81-86
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    • 2010
  • The mechanisms of selective oxidation of intercritically annealed CMnAl TRIP steels in a Continuous Galvanizing Line (GCL) were studied by cross-sectional observation of the surface and sub-surface regions by means of High Resolution Transmission Electron Microscopy (HR-TEM). The selective oxidation and nitriding of an intercritically annealed CMnAl TRIP steel in a controlled dew point 10%$H_2+N_2$ atmosphere resulted in the formation of c-xMnO.$MnO_2$ (1${\leq}$x<3) and c-xMnO.$Al_2O_3$ ($x{\geq}1$) particles on the steel surface. Single crystal c-xMnO.$SiO_2$ ($2{\leq}x{\leq}4$) oxide particles were also observed on the surface. A thin film of crystalline c-xMnO.$SiO_2$ (2${\leq}$x<3) and c-xMnO.$Al_2O_3$ ($x{\geq}1$) was present between these particles. In the sub-surface region, internal oxidation, nitriding and intermetallic compound formation were observed. In the first region, large crystalline c-xMnO.$SiO_2$ ($1{\geq}x{\geq}2$) and c-xMnO.$Al_2O_3$ ($x{\geq}1$) oxides particles were present. In the second region, c-AlN particles were observed, and in a third region, small $MnAl_x$ (x>1) intermetallic compound particles were observed.

On the Genesis of Okbang Tungsten Deposits (옥방(玉房) 중석광상(重石鑛床)의 성인(成因)에 관(關)한 연구(硏究) -특(特)히 남부광체(南部鑛體)에 대(對)하여-)

  • Youn, Jeung Su
    • Economic and Environmental Geology
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    • v.12 no.4
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    • pp.181-195
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    • 1979
  • The Nambu orebodies of the Okbang tungsten mine are hosted in the Precambrian amphibolite and Weonnam formation. These orebodies can be classified into two types; The scheelite-bearing ore vein occurring in the amphibolite (the Nambu 1, 2 adits) and tungsten-bearing quartz vein along the contact between the amphibolite and the Weonnam formation (the Young-ho, -1, -2, -3 levels). The scheelite-bearing ore vein in the amphilbolite is discontinuous, narrow, and highly irregular in geometry, occurring only within the amphibolite with which of the vein is graduational. Based on these feature of the mode of occurrence, the origin of this ore type might be attributed to a potential segregation of tungsten ore fluid in situ from hornblenditic basic magma of the host rock. Tungsten-bearing quartz vein, however, is considered to have deposited along the N30-60E trending fractures as a later hypothermal vein after the hornblendite was emplaced. The principal ore mineral is scheelite with minor amount of wolframite, and the gangue minerals are quartz, and small amounts of fluorite, pyrrhotite, chalcopyrite and calcite. Fluid inclusion study of minerals from the Nambu orebody reveals that the fluids in fluorite of the scheelite-bearning ore vein attained a temperature range of $208{\sim}256^{\circ}C$ and those in quartz from the tungsten-bearing quartz vein a temperature range of $220{\sim}357^{\circ}C$. The real formation temperatures can be somewhat higher than filling temperatures, if pressure correction is made. Chemical analysis of 8 amphibolitc samples on major and some trace elements indicate that the amphibolite is igneous origin. On a Niggli diagram (al-alk)versus c, the analytical values are plotted on an igneous field, and on a Niggli diagram mg versus c they follow a karroo igneous trend line. According to the Ba, Cr, and Ni versus Niggli mg plots suggested by Leake (1964), Okbang amphibolite fall outside a pelitic field and compare favorably with his plots form ortho-amphibolites. Analitical values of $MoO_3$ of 8 samples of scheelite minerals from the Nambu orebody indicate that the tungsten-bearing quartz vein (type n) of Nambu orebody shows a range from 1. 69% to 4.38% which is higher than 0.94%~3.25% $MoO_3$ for the scheelite-bearing ore vein (type I). This fact indicates that the type II was deposited in a lower $fO_2/higher$ $fO_2$ environment and under lower temperature than the type I. Analysis of major components $WO_3$, MnO, and FeO of 6 samples of wolframite from the type II veins revealed that they contain 73.35~76.2% $WO_3$, 7.94~11.63% MnO, and 10.53~14.82% FeO. MnO/FeO ratios of wolframite shows the range of 0.85~1.17 which suggests a slightly higher temperature type of deposits than other major tungsten deposits in the country.

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