• Title/Summary/Keyword: pyrophosphate

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Upregulation of thiamine (vitamin B1) biosynthesis gene upon stress application in Anabaena sp. and Nannochloropsis oculata

  • Fern, Lee Li;Abidin, Aisamuddin Ardi Zainal;Yusof, Zetty Norhana Balia
    • Journal of Plant Biotechnology
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    • v.44 no.4
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    • pp.462-471
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    • 2017
  • Thiamine pyrophosphate (TPP), the active form of thiamine is a cofactor for enzymes involved in central metabolism pathways. However, it is also known to have a role as a stress signaling molecule in response to environmental changes. Anabaena sp. and N. oculata are microorganisms which are abundantly found in Malaysia's freshwater and marine ecosystem. However, not much studies have been done especially in regards to thiamine biosynthesis. This work aimed to amplify of gene transcripts coding for thiamine biosynthesis enzymes besides looking at the expression of thiamine biosynthesis genes upon stress application. Various stress inducers were applied to the cultures and RNA was extracted at different time points. The first two genes, ThiC and ThiG/Thi4 encoding enzymes of the pyrimidine and thiazole branch respectively in the thiamine biosynthesis pathway were identified and amplified. The expression of the genes were analysed via RT-PCR and the intensity of bands were analysed using ImageJ software. The results showed up to 4-fold increase in the expression of ThiC and ThiG gene transcript as compared to control sample in Anabaena sp. ThiC gene in N. oculata showed an expression of 6-fold higher as compared to control sample. In conclusion, stresses induced the expression of the gene coding for one of the most important enzymes in thiamine biosynthesis pathway. This is an agreement with the hypothesis that overexpression of thiamine is crucial in assisting plants to combat abiotic stresses.

Characteristics of Thiamine Uptake by the BeWo Human Trophoblast Cell Line

  • Keating, Elisa;Lemos, Clara;Azevedo, Isabel;Martel, Fatima
    • BMB Reports
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    • v.39 no.4
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    • pp.383-393
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    • 2006
  • Little is known concerning the mechanisms responsible for the transplacental transfer of thiamine. So, the aim of this work was to characterize the placental uptake of thiamine from the maternal circulation, by determining the characteristics of $^3H$-thiamine uptake by a human trophoblast cell line (BeWo). Uptake of $^3H$-thiamine (50-100 nM) by BeWo cells was: 1) temperature-dependent and energy-independent; 2) pH-dependent (uptake increased as the extracellular medium pH decreased); 3) $Na^+$-dependent and $Cl^-$-independent; 4) not inhibited by the thiamine structural analogs amprolium, oxythiamine and thiamine pyrophosphate; 5) inhibited by the unrelated organic cations guanidine, N-methylnicotinamide, tetraethylammonium, clonidine and cimetidine; 6) inhibited by the organic cation serotonin, and by two selective inhibitors of the serotonin plasmalemmal transporter (hSERT), fluoxetine and desipramine. We conclude that $^3H$-thiamine uptake by BeWo cells seems to occur through a process distinct from thiamine transporter-1 (hThTr-1) and thiamine transporter-2 (hThTr-2). Rather, it seems to involve hSERT. Moreover, chronic (48 h) exposure of cells to caffeine ($1\;{\mu}M$) stimulated and chronic exposure to xanthohumol and iso-xanthohumol (1 and $0.1\;{\mu}M$, respectively) inhibited $^3H$-thiamine uptake, these effects being not mediated through modulation of the expression levels of either hThTr-1 or hSERT mRNA.

Studies on the Stability of $Methylenediphosphonate-^{99m}Tc$ (Methylenediphosphonate $(MDP)-^{99m}Tc$의 안정도(安定度)에 관(關)한 연구(硏究))

  • Kim, Jae-Rok;Awh, Ok-Doo;Park, Kwung-Bae;Koo, Hyeon-Sook
    • The Korean Journal of Nuclear Medicine
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    • v.16 no.2
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    • pp.81-84
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    • 1982
  • To evaluate the in-vivo and in-vitro stability of methylene diphosphonate $(MDP)-^{99m}Tc$, relative ligand exchange rates in phosphate buffer between $MDP-^{99m}Tc$ and human serum albumin (HSA), and between pyrophosphate$(PYP)-^{99m}Tc$ and HSA have been measured. Gel permeation chromatography has also been carried out to estimate relative rates of ligand exchange between polysaccharide and the $^{99m}Tc-bone$ agents. The in-vitro stability was further correlated with its specific radioactivity. The results indicated that the $MDP-^{99m}Tc$ is more stable than the $PYP-^{99m}Tc$ but less stable than $MIDA-^{99m}Tc$, $HIDA-^{99m}Tc$, and $DTPA-^{99m}Tc$ etc. in refering to other data. In stability point of view, $MDP-^{99m}Tc$ is considered to be a better bone agent than $PYP-^{99m}Tc$ since $MDP-^{99m}Tc$ can be accumulated at bone by a smooth transfer of $^{99m}Tc$ to hydroxyapatite, whereas in case of $PYP-^{99m}Tc$, most of the $^{99m}Tc$ is transferred to HSA before arriving to the hydroxyapatite.

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Preparation and Behavior of High Performance Alkaline Cleaning Agents for Steel (철강용 고성능 알칼리세정제의 제조 및 세정거동)

  • Park, Hong-Soo;Jo, Hye-Jin;Shim, Il-Woo;You, Hyuk-Jae;Kim, Myung-Soo;Hahm, Hyun-Sik
    • Journal of the Korean Applied Science and Technology
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    • v.22 no.1
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    • pp.15-20
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    • 2005
  • $Na_2CO_3$. Sodium orthosilicate (Na-OSi), Tetronix T-701 (T-701), Na-dioctyl sulfosuccinate (303C), Newpol PE-68 (PE-68), MJU-100A, and tetrasodium pyrophosphate were blended to prepare high performance alkaline cleaning agents (ACASs). The results of cleaning test with steel specimen showed that ACAS-6 ($Na_2CO_3$ 50g/Na-OSi 35g/T-701 20g/303C 18g/PE-68 17g/MJU-100A 10g/TSPP 20g/ water 180g mixture) had a good cleaning power. The cleaning power for press-rust preventing oil was 98% and 99% degreasing at 4wt%, $70^{\circ}C$ and $90^{\circ}C$, respectively ; for quenching oil, the cleaning power of ACAS-6 was 91% degreasing at 4wt% and $70^{\circ}C$. The foam heights measured immediately after foaming by Ross & Miles method and Ross & Clark method at 6wt%, $60^{\circ}C$ were 18mm and 65mm, respectively. It was concluded that ACAS-6 had a good low foaming cleaning agent.

Influence of High Light and Nitrate Deprivation on the Carotenoid Biosynthesis in Haematococcus pluvialis (고광도와 질소 결핍이 Haematococcus pluvialis의 색소 생합성에 미치는 영향)

  • Yun, Ji-Hyun;Kwak, In-Kyu;Jin, Eon-Seon
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.292-297
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    • 2007
  • The unicellular green alga, Haematococcus pluvialis used as a biological production system for astaxanthin. It accumulates large amounts of the red ketocarotenoid astaxanthin when exposed to various environmental stress such as active oxygen species and high light intensities. To induce astaxanthin biosynthesis of H. pluvialis, cells were incubated in either nitrate free at $25^{\circ}C$ under continuous high light intensity ($1,000\;{\mu}mol$ photons $m^{-2}s^{-1}$) for 2 days or high light stress only. Expressions of astaxanthin biosynthetic genes such as carotenoid hydroxylase, IPP isomerase and ${\beta}$-carotene ketolase were monitored under different culture conditions by using real time RT-PCR. All the subjected genes increased their expression under highlight and N-deprivation condition where a large amount of astaxanthin was accumulated.

A Protein Tyrosine Phosphatase Inhibitor, Pervanadate, Inhibits Angiotensin II-Induced β-Arrestin Cleavage

  • Jang, Sei-Heon;Hwang, Si Ae;Kim, Mijin;Yun, Sung-Hae;Kim, Moon-Sook;Karnik, Sadashiva S.;Lee, ChangWoo
    • Molecules and Cells
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    • v.28 no.1
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    • pp.25-30
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    • 2009
  • ${\beta}$-Arrestins turn off G protein-mediated signals and initiate distinct G protein-independent signaling pathways. We previously demonstrated that angiotensin $AT_1$ receptorbound ${\beta}$-arrestin 1 is cleaved after $Phe^{388}$ upon angiotensin II stimulation. The mechanism and signaling pathway of angiotensin II-induced ${\beta}$-arrestin cleavage remain largely unknown. Here, we show that protein Tyr phosphatase activity is involved in the regulation of ${\beta}$-arrestin 1 cleavage. Tagging of green fluorescent protein (GFP) either to the N-terminus or C-terminus of ${\beta}$-arrestin 1 induced conformational changes and the cleavage of ${\beta}$-arrestin 1 without angiotensin $AT_1$ receptor activation. Orthovanadate and molybdate, inhibitors of protein Tyr phosphatase, attenuated the cleavage of C-terminal GFP-tagged ${\beta}$-arrestin 1 in vitro. The inhibitory effects of okadaic acid and pyrophosphate, which are inhibitors of protein Ser/Thr phosphatase, were less than those of protein Tyr phosphatase inhibitors. Cell-permeable pervanadate inhibited angiotensin II-induced cleavage of ${\beta}$-arrestin 1 in COS-1 cells. Our findings suggest that Tyr phosphorylation signaling is involved in the regulation of angiotensin II-induced ${\beta}$-arrestin cleavage.

Optimization of the Processing Conditions for the Production of Cooked Pork Sausage as a Ready-to-Serve Product

  • Ohba K.;Livera J.R.J.;Seneviratne R.W.;Serjmyadag D.;Shimada K.;Fukushima M.;Han Kyu-Ho;Lee Chi-Ho;Sekikawa M.
    • Food Science of Animal Resources
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    • v.26 no.1
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    • pp.15-19
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    • 2006
  • The aim of this study was to determine the best processing conditions for producing of dried lean pork as a ready-to-serve product without using large-scale machines. Lean pork sausage was produced using 1.27% sodium chloride, 0.075% sodium polyphosphate, 0.06% sodium ascorbate, 0.075% sodium pyrophosphate, 0.009% sodium nitrite, 0.009% dextrin, 0.11% sodium glutamate and 1.4% spice mixture. The most appropriate slice thickness for drying was examined by slicing the sausage at a 0.5, 1 and 2 cm thickness. The drying temperatures were determined by drying the sausage slices at 35, 48 and $68^{\circ}$. The total drying period was for 12 hr, In order to examine the ability of this process to sterilize the pork, the raw meat materials were inoculated with Escherichia coli (E. coli). The optimal conditions for producing lean pork sausages were a 2 cm slice thickness and drying temperature of $68^{\circ}C$ for 12 hr, The moisture content water activity, color, hardness and pH were measured in the dried product. The product had a moisture content of 47.5% and a water activity of 0.93. There was a 47.7% percentage reduction in moisture. The dried product tested negative for E. coli even though the raw meat materials been inoculated with E. coli.

Effects of Some Sodium Phosphates as Auxiliary Agents for Softening Hard Water to Degum Silk (인산염이 견사련용수의 경수연화에 미치는 영향)

  • 신봉섭;남중희
    • Journal of Sericultural and Entomological Science
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    • v.36 no.1
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    • pp.37-43
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    • 1994
  • The effects of some sodium phosphates as auxiliary agents were studied on the blockade of hardness for silk degumming. In this work, four kinds of sodium phosphates were tested and the results were obtained through masking effects of metallic ions, difference of pH value and boil-off ratio. The degumming of calcium ingredient was analyzed by means of atomic absorption spectrophotometer and degumming test of cocoon shell was performed in the presence of calcium ingredient and sodium phosphates added to soap solution. In the view of the effects of sodium phosphates on calcium hardness, tetrasodium pyrophosphate(TSPP) and sodium phosphate dibasic(SPD) masked calcium ions more than sodium phosphate monobasic(SPM) and sodium hexametaphosphate(SHP). SHP and TSPP have excellent abilities of masking ferrous ions. The pH values of TSPP solution is higher than others, but lower than soap solution. The pH values were differently measured one another among the sodium phosphates but the boil-off ratio was increased in case of sodium phosphate with high pH value.

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Immunochemical Studies of Starfish Gangliosides: Production of Monoclonal Antibody against AG-2, the Major Ganglioside of Starfish Acanthaster planci, and Detecting Its Distribution in Tissues by TLC Immunostaining

  • Miyamoto, Tomofumi;Yamamoto, Atsushi;Sakai, Maki;Tanaka, Hiroyuki;Shoyama, Yukihiro;Higuchi, Ryuichi
    • Journal of Marine Bioscience and Biotechnology
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    • v.1 no.4
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    • pp.298-304
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    • 2006
  • In this study, we establish a thin-layer chromatography (TLC) immunostaining method for detecting starfish gangliosides. A new monoclonal antibody (MAb) against AG-2, the major gangliosides molecular species of Acanthaster planci, was produced by fusing hybridoma with splenocytes immunized to liposomal AG-2. BALB/c male mice were injected with liposomal AG-2 antigen, and immunized. Their splenocytos were isolated and fused with hypoxanthine-aminopterine-thimidine (HAT)-sensitive mouse myeloma cells. Hybridomas producing MAb reactive to AG-2 were cloned using the limited dilution method. Established hybridomas were cultured in eRDF medium. Crude MAb produced from clone 8D4 was purified with a magnesium pyrophosphate column. Enzyme immunoassay and TLC immunostaining of AG-2 were performed using the purified MAb. Structurally related gangliosides did not cross-react with anti-AG-2 antibodies. The detection limit of TLC immunostaining was 50 ng of AG-2. The newly established immunostaining method was further developed for detecting AG-2 distribution and qualitative analysis in tissues and/or organs. Our results show that the majority of AG-2 is present in the stomach of male A. planci, while AG-2 is distributed not only in the stomach but also in the the pyloric caeca of female A. planci.

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Effects of Treatment Method and Environmental Factors on the Bacteriostatic Activity Condensed Phosphates (처리조건이나 환경요인이 중합인산염의 항균력에 미치는 영향)

  • CHANG Dong-Suck;LEE Tai-Seek
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.23 no.5
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    • pp.394-400
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    • 1990
  • In the previous paper, we reported that the bacteriostatic effect of condensed phosphate. The present study was intended to observe influence of various environmental factors on the bacteriostatic effect of condensed phosphates in the laboratory media, in order to get the information on the possibility to use the phosphate as food preservative. Bacteriostatic effect of sodium polyphosphate was not reduced by the heating at 100 for 1 hour, but it was considerably decreased by heating at $121^{\circ}C$ for 15 min and the phosphate sensitivity of bacteria was increased by freezing and heating. On the other hand, the strong bacteriostatic activity of condensed phosphate was observed below pH 6.5 in nutrient broth culture, and the activity was decreased by the addition of $CaCl_2$, KCl and $MgSO_4$.

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