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Sample records for aldose reductase ar

  1. Aldose reductase mediates retinal microglia activation.

    PubMed

    Chang, Kun-Che; Shieh, Biehuoy; Petrash, J Mark

    2016-04-29

    Retinal microglia (RMG) are one of the major immune cells in charge of surveillance of inflammatory responses in the eye. In the absence of an inflammatory stimulus, RMG reside predominately in the ganglion layer and inner or outer plexiform layers. However, under stress RMG become activated and migrate into the inner nuclear layer (INL) or outer nuclear layer (ONL). Activated RMG in cell culture secrete pro-inflammatory cytokines in a manner sensitive to downregulation by aldose reductase inhibitors. In this study, we utilized CX3CR1(GFP) mice carrying AR mutant alleles to evaluate the role of AR on RMG activation and migration in vivo. When tested on an AR(WT) background, IP injection of LPS induced RMG activation and migration into the INL and ONL. However, this phenomenon was largely prevented by AR inhibitors or in AR null mice, or was exacerbated in transgenic mice that over-express AR. LPS-induced increases in ocular levels of TNF-α and CX3CL-1 in WT mice were substantially lower in AR null mice or were reduced by AR inhibitor treatment. These studies demonstrate that AR expression in RMG may contribute to the proinflammatory phenotypes common to various eye diseases such as uveitis and diabetic retinopathy. PMID:27033597

  2. Aldose reductase mediates retinal microglia activation.

    PubMed

    Chang, Kun-Che; Shieh, Biehuoy; Petrash, J Mark

    2016-04-29

    Retinal microglia (RMG) are one of the major immune cells in charge of surveillance of inflammatory responses in the eye. In the absence of an inflammatory stimulus, RMG reside predominately in the ganglion layer and inner or outer plexiform layers. However, under stress RMG become activated and migrate into the inner nuclear layer (INL) or outer nuclear layer (ONL). Activated RMG in cell culture secrete pro-inflammatory cytokines in a manner sensitive to downregulation by aldose reductase inhibitors. In this study, we utilized CX3CR1(GFP) mice carrying AR mutant alleles to evaluate the role of AR on RMG activation and migration in vivo. When tested on an AR(WT) background, IP injection of LPS induced RMG activation and migration into the INL and ONL. However, this phenomenon was largely prevented by AR inhibitors or in AR null mice, or was exacerbated in transgenic mice that over-express AR. LPS-induced increases in ocular levels of TNF-α and CX3CL-1 in WT mice were substantially lower in AR null mice or were reduced by AR inhibitor treatment. These studies demonstrate that AR expression in RMG may contribute to the proinflammatory phenotypes common to various eye diseases such as uveitis and diabetic retinopathy.

  3. Aldose Reductase, Oxidative Stress, and Diabetic Mellitus

    PubMed Central

    Tang, Wai Ho; Martin, Kathleen A.; Hwa, John

    2012-01-01

    Diabetes mellitus (DM) is a complex metabolic disorder arising from lack of insulin production or insulin resistance (Diagnosis and classification of diabetes mellitus, 2007). DM is a leading cause of morbidity and mortality in the developed world, particularly from vascular complications such as atherothrombosis in the coronary vessels. Aldose reductase (AR; ALR2; EC 1.1.1.21), a key enzyme in the polyol pathway, catalyzes nicotinamide adenosine dinucleotide phosphate-dependent reduction of glucose to sorbitol, leading to excessive accumulation of intracellular reactive oxygen species (ROS) in various tissues of DM including the heart, vasculature, neurons, eyes, and kidneys. As an example, hyperglycemia through such polyol pathway induced oxidative stress, may have dual heart actions, on coronary blood vessel (atherothrombosis) and myocardium (heart failure) leading to severe morbidity and mortality (reviewed in Heather and Clarke, 2011). In cells cultured under high glucose conditions, many studies have demonstrated similar AR-dependent increases in ROS production, confirming AR as an important factor for the pathogenesis of many diabetic complications. Moreover, recent studies have shown that AR inhibitors may be able to prevent or delay the onset of cardiovascular complications such as ischemia/reperfusion injury, atherosclerosis, and atherothrombosis. In this review, we will focus on describing pivotal roles of AR in the pathogenesis of cardiovascular diseases as well as other diabetic complications, and the potential use of AR inhibitors as an emerging therapeutic strategy in preventing DM complications. PMID:22582044

  4. Aldose reductase, oxidative stress, and diabetic mellitus.

    PubMed

    Tang, Wai Ho; Martin, Kathleen A; Hwa, John

    2012-01-01

    Diabetes mellitus (DM) is a complex metabolic disorder arising from lack of insulin production or insulin resistance (Diagnosis and classification of diabetes mellitus, 2007). DM is a leading cause of morbidity and mortality in the developed world, particularly from vascular complications such as atherothrombosis in the coronary vessels. Aldose reductase (AR; ALR2; EC 1.1.1.21), a key enzyme in the polyol pathway, catalyzes nicotinamide adenosine dinucleotide phosphate-dependent reduction of glucose to sorbitol, leading to excessive accumulation of intracellular reactive oxygen species (ROS) in various tissues of DM including the heart, vasculature, neurons, eyes, and kidneys. As an example, hyperglycemia through such polyol pathway induced oxidative stress, may have dual heart actions, on coronary blood vessel (atherothrombosis) and myocardium (heart failure) leading to severe morbidity and mortality (reviewed in Heather and Clarke, 2011). In cells cultured under high glucose conditions, many studies have demonstrated similar AR-dependent increases in ROS production, confirming AR as an important factor for the pathogenesis of many diabetic complications. Moreover, recent studies have shown that AR inhibitors may be able to prevent or delay the onset of cardiovascular complications such as ischemia/reperfusion injury, atherosclerosis, and atherothrombosis. In this review, we will focus on describing pivotal roles of AR in the pathogenesis of cardiovascular diseases as well as other diabetic complications, and the potential use of AR inhibitors as an emerging therapeutic strategy in preventing DM complications. PMID:22582044

  5. Aldose reductase inhibitory activity of compounds from Zea mays L.

    PubMed

    Kim, Tae Hyeon; Kim, Jin Kyu; Kang, Young-Hee; Lee, Jae-Yong; Kang, Il Jun; Lim, Soon Sung

    2013-01-01

    Aldose reductase (AR) inhibitors have a considerable therapeutic potential against diabetes complications and do not increase the risk of hypoglycemia. Through bioassay-guided fractionation of an EtOH extract of the kernel from purple corn (Zea mays L.), 7 nonanthocyanin phenolic compounds (compound 1-7) and 5 anthocyanins (compound 8-12) were isolated. These compounds were investigated by rat lens aldose reductase (RLAR) inhibitory assays. Kinetic analyses of recombinant human aldose reductase (rhAR) were performed, and intracellular galactitol levels were measured. Hirsutrin, one of 12 isolated compounds, showed the most potent RLAR inhibitory activity (IC(50), 4.78 μ M). In the kinetic analyses using Lineweaver-Burk plots of 1/velocity and 1/substrate concentration, hirsutrin showed competitive inhibition against rhAR. Furthermore, hirsutrin inhibited galactitol formation in rat lens and erythrocytes sample incubated with a high concentration of galactose; this finding indicates that hirsutrin may effectively prevent osmotic stress in hyperglycemia. Therefore, hirsutrin derived from Zea mays L. may be a potential therapeutic agent against diabetes complications. PMID:23586057

  6. Structure of aldose reductase from Giardia lamblia

    PubMed Central

    Ferrell, M.; Abendroth, J.; Zhang, Y.; Sankaran, B.; Edwards, T. E.; Staker, B. L.; Van Voorhis, W. C.; Stewart, L. J.; Myler, P. J.

    2011-01-01

    Giardia lamblia is an anaerobic aerotolerant eukaryotic parasite of the intestines. It is believed to have diverged early from eukarya during evolution and is thus lacking in many of the typical eukaryotic organelles and biochemical pathways. Most conspicuously, mitochondria and the associated machinery of oxidative phosphorylation are absent; instead, energy is derived from substrate-level phosphorylation. Here, the 1.75 Å resolution crystal structure of G. lamblia aldose reductase heterologously expressed in Escherichia coli is reported. As in other oxidoreductases, G. lamblia aldose reductase adopts a TIM-barrel conformation with the NADP+-binding site located within the eight β-strands of the interior. PMID:21904059

  7. Studies on aldose reductase inhibitors from natural products. IV. Constituents and aldose reductase inhibitory effect of Chrysanthemum morifolium, Bixa orellana and Ipomoea batatas.

    PubMed

    Terashima, S; Shimizu, M; Horie, S; Morita, N

    1991-12-01

    The hot water extracts of Chrysanthemum morifolium, Bixa orellana and Ipomoea batatas, were found to have potent inhibitory activity towards lens aldose reductase (AR). Ellagic acid (4) was isolated from C. morifolium and I. batatas, isoscutellarein (7) from B. orellana and 3,5-dicaffeoylquinic acid (10) from I. batatas, respectively, as potent inhibitors. PMID:1814628

  8. B-factor Analysis and Conformational Rearrangement of Aldose Reductase.

    PubMed

    Balendiran, Ganesaratnam K; Pandian, J Rajendran; Drake, Evin; Vinayak, Anubhav; Verma, Malkhey; Cascio, Duilio

    2014-01-01

    The NADPH-dependent reduction of glucose reaction that is catalyzed by Aldose Reductase (AR) follows a sequential ordered kinetic mechanism in which the co-factor NADPH binds to the enzyme prior to the aldehyde substrate. The kinetic/structural experiments have found a conformational change involving a hinge-like movement of a surface loop (residues 213-224) which is anticipated to take place upon the binding of the diphosphate moiety of NADPH. The reorientation of this loop, expected to permit the release of NADP(+), represents the rate-limiting step of the catalytic mechanism. This study reveals: 1) The Translation/Libration/Screw (TLS) analysis of absolute B-factors of apo AR crystal structures indicates that the 212-224 loop might move as a rigid group. 2) Residues that make the flexible loop slide in the AR binary and ternary complexes. 3) The normalized B-factors separate this segment into three different clusters with fewer residues.

  9. Crystallization and preliminary X-ray diffraction analysis of maize aldose reductase

    SciTech Connect

    Kiyota, Eduardo; Sousa, Sylvia Morais de; Santos, Marcelo Leite dos; Costa Lima, Aline da; Menossi, Marcelo; Yunes, José Andrés; Aparicio, Ricardo

    2007-11-01

    Preliminary X-ray diffraction studies of apo maize aldose reductase at 2.0 Å resolution are reported. Maize aldose reductase (AR) is a member of the aldo-keto reductase superfamily. In contrast to human AR, maize AR seems to prefer the conversion of sorbitol into glucose. The apoenzyme was crystallized in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 47.2, b = 54.5, c = 100.6 Å and one molecule in the asymmetric unit. Synchrotron X-ray diffraction data were collected and a final resolution limit of 2.0 Å was obtained after data reduction. Phasing was carried out by an automated molecular-replacement procedure and structural refinement is currently in progress. The refined structure is expected to shed light on the functional/enzymatic mechanism and the unusual activities of maize AR.

  10. The inhibitory activity of aldose reductase in vitro by constituents of Garcinia mangostana Linn.

    PubMed

    Fatmawati, Sri; Ersam, Taslim; Shimizu, Kuniyoshi

    2015-01-15

    We investigated aldose reductase inhibition of Garcinia mangostana Linn. from Indonesia. Dichloromethane extract of the root bark of this tree was found to demonstrate an IC50 value of 11.98 µg/ml for human aldose reductase in vitro. From the dichloromethane fraction, prenylated xanthones were isolated as potent human aldose reductase inhibitors. We discovered 3-isomangostin to be most potent against aldose reductase, with an IC50 of 3.48 µM.

  11. Autoimmunity in Membranous Nephropathy Targets Aldose Reductase and SOD2

    PubMed Central

    Prunotto, Marco; Carnevali, Maria Luisa; Candiano, Giovanni; Murtas, Corrado; Bruschi, Maurizio; Corradini, Emilia; Trivelli, Antonella; Magnasco, Alberto; Petretto, Andrea; Santucci, Laura; Mattei, Silvia; Gatti, Rita; Scolari, Francesco; Kador, Peter; Allegri, Landino

    2010-01-01

    Glomerular targets of autoimmunity in human membranous nephropathy are poorly understood. Here, we used a combined proteomic approach to identify specific antibodies against podocyte proteins in both serum and glomeruli of patients with membranous nephropathy (MN). We detected specific anti–aldose reductase (AR) and anti–manganese superoxide dismutase (SOD2) IgG4 in sera of patients with MN. We also eluted high titers of anti-AR and anti-SOD2 IgG4 from microdissected glomeruli of three biopsies of MN kidneys but not from biopsies of other glomerulonephritides characterized by IgG deposition (five lupus nephritis and two membranoproliferative glomerulonephritis). We identified both antigens in MN biopsies but not in other renal pathologies or normal kidney. Confocal and immunoelectron microscopy (IEM) showed co-localization of anti-AR and anti-SOD2 with IgG4 and C5b-9 in electron-dense podocyte immune deposits. Preliminary in vitro experiments showed an increase of SOD2 expression on podocyte plasma membrane after treatment with hydrogen peroxide. In conclusion, our data support AR and SOD2 as renal antigens of human MN and suggest that oxidative stress may drive glomerular SOD2 expression. PMID:20150532

  12. Aldose reductase expression as a risk factor for cataract.

    PubMed

    Snow, Anson; Shieh, Biehuoy; Chang, Kun-Che; Pal, Arttatrana; Lenhart, Patricia; Ammar, David; Ruzycki, Philip; Palla, Suryanarayana; Reddy, G Bhanuprakesh; Petrash, J Mark

    2015-06-01

    Aldose reductase (AR) is thought to play a role in the pathogenesis of diabetic eye diseases, including cataract and retinopathy. However, not all diabetics develop ocular complications. Paradoxically, some diabetics with poor metabolic control appear to be protected against retinopathy, while others with a history of excellent metabolic control develop severe complications. These observations indicate that one or more risk factors may influence the likelihood that an individual with diabetes will develop cataracts and/or retinopathy. We hypothesize that an elevated level of AR gene expression could confer higher risk for development of diabetic eye disease. To investigate this hypothesis, we examined the onset and severity of diabetes-induced cataract in transgenic mice, designated AR-TG, that were either heterozygous or homozygous for the human AR (AKR1B1) transgene construct. AR-TG mice homozygous for the transgene demonstrated a conditional cataract phenotype, whereby they developed lens vacuoles and cataract-associated structural changes only after induction of experimental diabetes; no such changes were observed in AR-TG heterozygotes or nontransgenic mice with or without experimental diabetes induction. We observed that nondiabetic AR-TG mice did not show lens structural changes even though they had lenticular sorbitol levels almost as high as the diabetic AR-TG lenses that showed early signs of cataract. Over-expression of AR led to increases in the ratio of activated to total levels of extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal (JNK1/2), which are known to be involved in cell growth and apoptosis, respectively. After diabetes induction, AR-TG but not WT controls had decreased levels of phosphorylated as well as total ERK1/2 and JNK1/2 compared to their nondiabetic counterparts. These results indicate that high AR expression in the context of hyperglycemia and insulin deficiency may constitute a risk factor that could predispose the

  13. Aldose reductase expression as a risk factor for cataract.

    PubMed

    Snow, Anson; Shieh, Biehuoy; Chang, Kun-Che; Pal, Arttatrana; Lenhart, Patricia; Ammar, David; Ruzycki, Philip; Palla, Suryanarayana; Reddy, G Bhanuprakesh; Petrash, J Mark

    2015-06-01

    Aldose reductase (AR) is thought to play a role in the pathogenesis of diabetic eye diseases, including cataract and retinopathy. However, not all diabetics develop ocular complications. Paradoxically, some diabetics with poor metabolic control appear to be protected against retinopathy, while others with a history of excellent metabolic control develop severe complications. These observations indicate that one or more risk factors may influence the likelihood that an individual with diabetes will develop cataracts and/or retinopathy. We hypothesize that an elevated level of AR gene expression could confer higher risk for development of diabetic eye disease. To investigate this hypothesis, we examined the onset and severity of diabetes-induced cataract in transgenic mice, designated AR-TG, that were either heterozygous or homozygous for the human AR (AKR1B1) transgene construct. AR-TG mice homozygous for the transgene demonstrated a conditional cataract phenotype, whereby they developed lens vacuoles and cataract-associated structural changes only after induction of experimental diabetes; no such changes were observed in AR-TG heterozygotes or nontransgenic mice with or without experimental diabetes induction. We observed that nondiabetic AR-TG mice did not show lens structural changes even though they had lenticular sorbitol levels almost as high as the diabetic AR-TG lenses that showed early signs of cataract. Over-expression of AR led to increases in the ratio of activated to total levels of extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal (JNK1/2), which are known to be involved in cell growth and apoptosis, respectively. After diabetes induction, AR-TG but not WT controls had decreased levels of phosphorylated as well as total ERK1/2 and JNK1/2 compared to their nondiabetic counterparts. These results indicate that high AR expression in the context of hyperglycemia and insulin deficiency may constitute a risk factor that could predispose the

  14. Aldose reductase expression as a risk factor for cataract

    PubMed Central

    Snow, Anson; Shieh, Biehuoy; Chang, Kun-Che; Pal, Arttatrana; Lenhart, Patricia; Ammar, David; Ruzycki, Philip; Palla, Suryanarayana; Reddy, G. Bhanuprakesh; Petrash, J. Mark

    2015-01-01

    Aldose reductase (AR) is thought to play a role in the pathogenesis of diabetic eye diseases, including cataract and retinopathy. However, not all diabetics develop ocular complications. Paradoxically, some diabetics with poor metabolic control appear to be protected against retinopathy, while others with a history of excellent metabolic control develop severe complications. These observations indicate that one or more risk factors may influence the likelihood that an individual with diabetes will develop cataracts and/or retinopathy. We hypothesize that an elevated level of AR gene expression could confer higher risk for development of diabetic eye disease. To investigate this hypothesis, we examined the onset and severity of diabetes-induced cataract in transgenic mice, designated AR-TG, that were either heterozygous or homozygous for the human AR (AKR1B1) transgene construct. AR-TG mice homozygous for the transgene demonstrated a conditional cataract phenotype, whereby they developed lens vacuoles and cataract-associated structural changes only after induction of experimental diabetes; no such changes were observed in AR-TG heterozygotes or nontransgenic mice with or without experimental diabetes induction. We observed that nondiabetic AR-TG mice did not show lens structural changes even though they had lenticular sorbitol levels almost as high as the diabetic AR-TG lenses that showed early signs of cataract. Over-expression of AR led to increases in the ratio of activated to total levels of extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal (JNK1/2), which are known to be involved in cell growth and apoptosis respectively. After diabetes induction, AR-TG but not WT controls had decreased levels of phosphorylated as well as total ERK1/2 and JNK1/2 compared to their nondiabetic counterparts. These results indicate that high AR expression in the context of hyperglycemia and insulin deficiency may constitute a risk factor that could predispose the

  15. Isoquinoline alkaloids from Tinospora cordifolia inhibit rat lens aldose reductase.

    PubMed

    Patel, Mayurkumar B; Mishra, Shrihari

    2012-09-01

    The inhibitory activity of Tinospora cordifolia stem-derived alkaloids was evaluated against lens aldose reductase (AR) isolated from male Wistar rats. Anticataract potential of the alkaloids of T. cordifolia was evaluated in vitro in rat lenses, considering the activity of normal rat lenses as 100%. The biologically active constituents of T. cordifolia extract were characterized as the isoquinoline alkaloids, jatrorrhizine, palmatine and magnoflorine, by spectral analysis. The inhibitory effects varied with all chemicals and concentrations used. The inhibitory concentration (IC₅₀) values of jatrorrhizine, palmatine and magnoflorine are 3.23, 3.45 and 1.25 µg/mL respectively. The concentration of maximum activity was selected for its effect on galactose-induced polyol accumulation in vitro. The percentage inhibition of galactose-induced polyol accumulation was 62.6, 58.8 and 27.7% in the presence of jatrorrhizine, palmatine and magnoflorine, respectively. Magnoflorine may be useful as lead compounds and new agents for AR inhibition. PMID:22294283

  16. Aldose Reductase Inhibitory Activity of Compounds from  Zea mays L.

    PubMed Central

    Kim, Tae Hyeon; Kim, Jin Kyu; Kang, Young-Hee; Lee, Jae-Yong; Kang, Il Jun; Lim, Soon Sung

    2013-01-01

    Aldose reductase (AR) inhibitors have a considerable therapeutic potential against diabetes complications and do not increase the risk of hypoglycemia. Through bioassay-guided fractionation of an EtOH extract of the kernel from purple corn (Zea mays L.), 7 nonanthocyanin phenolic compounds (compound 1–7) and 5 anthocyanins (compound 8–12) were isolated. These compounds were investigated by rat lens aldose reductase (RLAR) inhibitory assays. Kinetic analyses of recombinant human aldose reductase (rhAR) were performed, and intracellular galactitol levels were measured. Hirsutrin, one of 12 isolated compounds, showed the most potent RLAR inhibitory activity (IC50, 4.78 μM). In the kinetic analyses using Lineweaver-Burk plots of 1/velocity and 1/substrate concentration, hirsutrin showed competitive inhibition against rhAR. Furthermore, hirsutrin inhibited galactitol formation in rat lens and erythrocytes sample incubated with a high concentration of galactose; this finding indicates that hirsutrin may effectively prevent osmotic stress in hyperglycemia. Therefore, hirsutrin derived from Zea mays L. may be a potential therapeutic agent against diabetes complications. PMID:23586057

  17. Aldose reductase inhibitors from the leaves of Myrciaria dubia (H. B. & K.) McVaugh.

    PubMed

    Ueda, H; Kuroiwa, E; Tachibana, Y; Kawanishi, K; Ayala, F; Moriyasu, M

    2004-11-01

    Ellagic acid (1) and its two derivatives, 4-O-methylellagic acid (2) and 4-(alpha-rhamnopyranosyl)ellagic acid (3) were isolated as inhibitors of aldose reductase (AR) from Myrciaria dubia (H. B. & K.) McVaugh. Compound 2 was the first isolated from the nature. Compound 3 showed the strongest inhibition against human recombinant AR (HRAR) and rat lens AR (RLAR). Inhibitory activity of compound 3 against HRAR (IC50 value = 4.1 x 10(-8) M) was 60 times more than that of quercetin (2.5 x 10(-6) M). The type of inhibition against HRAR was uncompetitive. PMID:15636180

  18. Rapid Identification of Aldose Reductase Inhibitory Compounds from Perilla frutescens

    PubMed Central

    Paek, Ji Hun; Shin, Kuk Hyun; Kang, Young-Hee; Lee, Jae-Yong; Lim, Soon Sung

    2013-01-01

    The ethyl acetate (EtOAc) soluble fraction of methanol extracts of Perilla frutescens (P. frutescens) inhibits aldose reductase (AR), the key enzyme in the polyol pathway. Our investigation of inhibitory compounds from the EtOAc soluble fraction of P. frutescens was followed by identification of the inhibitory compounds by a combination of HPLC microfractionation and a 96-well enzyme assay. This allowed the biological activities to be efficiently matched with selected HPLC peaks. Structural analyses of the active compounds were performed by LC-MSn. The main AR inhibiting compounds were tentatively identified as chlorogenic acid and rosmarinic acid by LC-MSn. A two-step high speed counter current chromatography (HSCCC) isolation method was developed with a solvent system of n-hexane-ethyl acetate-methanol-water at 1.5 : 5 : 1 : 5, v/v and 3 : 7 : 5 : 5, v/v. The chemical structures of the isolated compounds were determined by 1H- and 13C-nuclear magnetic resonance spectrometry (NMR). The main compounds inhibiting AR in the EtOAc fraction of methanol extracts of P. frutescens were identified as chlorogenic acid (2) (IC50 = 3.16 μM), rosmarinic acid (4) (IC50 = 2.77 μM), luteolin (5) (IC50 = 6.34 μM), and methyl rosmarinic acid (6) (IC50 = 4.03 μM). PMID:24308003

  19. Inhibition of aldose reductase by phenylethanoid glycoside isolated from the seeds of Paulownia coreana.

    PubMed

    Kim, Jin Kyu; Lee, Yeon Sil; Kim, Seon Ha; Bae, Young Soo; Lim, Soon Sung

    2011-01-01

    Aldose reductase (AR) inhibitors have considerable therapeutic potential against diabetic complications and do not increase the risk of hypoglycemia. Through bioassay-guided fractionation of the 70% acetone extract obtained from Paulownia coreana seeds, phenylpropanoid glycosides (compounds 1-4) and 5 phenolic compounds were isolated (compounds 5-9). Their structures were determined on the basis of spectroscopic analysis and comparison with reported data. All the isolates were subjected to in vitro bioassays to evaluate their inhibitory activities against recombinant human aldose reductase (rhAR) and sorbitol formation in human erythrocytes. Phenylethanoid glycosides showed more effective than the phenolic compounds in inhibiting rhAR. Among the compounds, isocampneoside II (3) was found to significantly inhibit rhAR with an IC(50) value of 9.72 µM. In kinetic analyses performed using Lineweaver-Burk plots of 1/velocity and 1/concentration of substrate, isocampneoside II (3) showed uncompetitive inhibition against rhAR. Furthermore, it inhibited sorbitol formation in a rat lens incubated with a high concentration of glucose; this finding indicated that isocampneoside II (3) may effectively prevent osmotic stress in hyperglycemia. Thus, the P. coreana-derived phenylethanoid glycoside isocampneoside II (3) may have a potential therapeutics against diabetic complications. PMID:21212537

  20. Synthesis of organic nitrates of luteolin as a novel class of potent aldose reductase inhibitors.

    PubMed

    Wang, Qi-Qin; Cheng, Ning; Zheng, Xiao-Wei; Peng, Sheng-Ming; Zou, Xiao-Qing

    2013-07-15

    Aldose reductase (AR) plays an important role in the design of drugs that prevent and treat diabetic complications. Aldose reductase inhibitors (ARIs) have received significant attentions as potent therapeutic drugs. Based on combination principles, three series of luteolin derivatives were synthesised and evaluated for their AR inhibitory activity and nitric oxide (NO)-releasing capacity in vitro. Eighteen compounds were found to be potent ARIs with IC50 values ranging from (0.099±0.008) μM to (2.833±0.102) μM. O(7)-Nitrooxyethyl-O(3'),O(4')-ethylidene luteolin (La1) showed the most potent AR inhibitory activity [IC50=(0.099±0.008) μM]. All organic nitrate derivatives released low concentrations of NO in the presence of l-cysteine. Structure-activity relationship studies suggested that introduction of an NO donor, protection of the catechol structure, and the ether chain of a 2-carbon spacer as a coupling chain on the luteolin scaffold all help increase the AR inhibitory activity of the resulting compound. This class of NO-donor luteolin derivatives as efficient ARIs offer a new concept for the development and design of new drug for preventive and therapeutic drugs for diabetic complications.

  1. 20(S)-Ginsenoside Rh2 as aldose reductase inhibitor from Panax ginseng.

    PubMed

    Fatmawati, Sri; Ersam, Taslim; Yu, Hongshan; Zhang, Chunzhi; Jin, Fengxie; Shimizu, Kuniyoshi

    2014-09-15

    The root of Panax ginseng C. A. Meyer (Araliaceae) is a well-known herbal medicine in East Asia. The major bioactive metabolites in this root are commonly identified as ginsenosides. A series of ginsenosides were determined for in vitro human recombinant aldose reductase. This Letter aims to clarify the structural requirement for aldose reductase inhibition. We discovered that only ginsenoside 20(S)-Rh2 showed potent against aldose reductase, with an IC50 of 147.3 μM. These results implied that the stereochemistry of the hydroxyl group at C-20 may play an important role in aldose reductase inhibition. An understanding of these requirements is considered necessary in order to develop a new type of aldose reductase inhibitor. Furthermore, P. ginseng might be an important herbal medicine in preventing diabetic complications.

  2. Aldose reductase catalysis and crystallography. Insights from recent advances in enzyme structure and function.

    PubMed

    Petrash, J M; Tarle, I; Wilson, D K; Quiocho, F A

    1994-08-01

    Enhanced metabolism of glucose via the polyol pathway may play an important role in the pathogenesis of diabetic retinopathy, neuropathy, and nephropathy. Aldose reductase catalyzes the NADPH-dependent conversion of glucose to sorbitol, the first step in the polyol pathway. Interruption of the polyol pathway by inhibition of aldose reductase holds considerable promise as a therapeutic measure to prevent or delay the onset and severity of these late complications of diabetes. Dramatic advances in our understanding of the molecular biology, enzymology, and three-dimensional structure of aldose reductase have occurred in recent years, providing new and challenging insights into the enzyme's catalytic mechanism. Recent developments in structure determination of aldose reductase and the implications for evaluation and development of aldose reductase inhibitors are summarized. PMID:8039602

  3. Aldose and aldehyde reductases : structure-function studies on the coenzyme and inhibitor-binding sites.

    SciTech Connect

    El-Kabbani, O.; Old, S. E.; Ginell, S. L.; Carper, D. A.; Biosciences Division; Monash Univ.; NIH

    1999-09-03

    PURPOSE: To identify the structural features responsible for the differences in coenzyme and inhibitor specificities of aldose and aldehyde reductases. METHODS: The crystal structure of porcine aldehyde reductase in complex with NADPH and the aldose reductase inhibitor sorbinil was determined. The contribution of each amino acid lining the coenzyme-binding site to the binding of NADPH was calculated using the Discover package. In human aldose reductase, the role of the non-conserved Pro 216 (Ser in aldehyde reductase) in the binding of coenzyme was examined by site-directed mutagenesis. RESULTS: Sorbinil binds to the active site of aldehyde reductase and is hydrogen-bonded to Trp 22, Tyr 50, His 113, and the non-conserved Arg 312. Unlike tolrestat, the binding of sorbinil does not induce a change in the side chain conformation of Arg 312. Mutation of Pro 216 to Ser in aldose reductase makes the binding of coenzyme more similar to that of aldehyde reductase. CONCLUSIONS: The participation of non-conserved active site residues in the binding of inhibitors and the differences in the structural changes required for the binding to occur are responsible for the differences in the potency of inhibition of aldose and aldehyde reductases. We report that the non-conserved Pro 216 in aldose reductase contributes to the tight binding of NADPH.

  4. Altered aldose reductase gene regulation in cultured human retinal pigment epithelial cells.

    PubMed Central

    Henry, D N; Del Monte, M; Greene, D A; Killen, P D

    1993-01-01

    Aldose reductase (AR2), a putative "hypertonicity stress protein" whose gene is induced by hyperosmolarity, protects renal medullary cells against the interstitial hyperosmolarity of antidiuresis by catalyzing the synthesis of millimolar concentrations of intracellular sorbitol from glucose. Although AR2 gene induction has been noted in a variety of renal and nonrenal cells subjected to hypertonic stress in vitro, the functional significance of AR2 gene expression in cells not normally exposed to a hyperosmolar milieu is not fully understood. The physiological impact of basal AR2 expression in such cells may be limited to hyperglycemic states in which AR2 promotes pathological polyol accumulation, a mechanism invoked in the pathogenesis of diabetic complications. Since AR2 overexpression in the retinal pigment epithelium has been associated with diabetic retinopathy, the regulation of AR2 gene expression and associated changes in sorbitol and myo-inositol were studied in human retinal pigment epithelial cells in culture. The relative abundance of aldehyde reductase (AR1) and AR2 mRNA was quantitated by filter hybridization of RNA from several human retinal pigment epithelial cell lines exposed to hyperglycemic and hyperosmolar conditions in vitro. AR2 but not AR1 mRNA was significantly increased some 11- to 18-fold by hyperosmolarity in several retinal pigment epithelial cell lines. A single cell line with a 15-fold higher basal level of AR2 mRNA than other cell lines tested demonstrated no significant increase in AR2 mRNA in response to hypertonic stress. This cell line demonstrated accelerated and exaggerated production of sorbitol and depletion of myo-inositol upon exposure to 20 mM glucose. Therefore, abnormal AR2 expression may enhance the sensitivity of cells to the biochemical consequences of hyperglycemia potentiating the development of diabetic complications. Images PMID:8349800

  5. Characterization of WY 14,643 and its Complex with Aldose Reductase

    PubMed Central

    Sawaya, Michael R.; Verma, Malkhey; Balendiran, Vaishnavi; Rath, Nigam P.; Cascio, Duilio; Balendiran, Ganesaratnam K.

    2016-01-01

    The peroxisome proliferator, WY 14,643 exhibits a pure non-competitive inhibition pattern in the aldehyde reduction and in alcohol oxidation activities of human Aldose reductase (hAR). Fluorescence emission measurements of the equilibrium dissociation constants, Kd, of oxidized (hAR•NADP+) and reduced (hAR•NADPH) holoenzyme complexes display a 2-fold difference between them. Kd values for the dissociation of WY 14,643 from the oxidized (hAR•NADP+•WY 14,643) and reduced (hAR•NADPH•WY 14,643) ternary complexes are comparable to each other. The ternary complex structure of hAR•NADP+•WY 14,643 reveals the first structural evidence of a fibrate class drug binding to hAR. These observations demonstrate how fibrate molecules such as WY 14,643, besides being valued as agonists for PPAR, also inhibit hAR. PMID:27721416

  6. Novel Role for Aldose Reductase in Mediating Acute Inflammatory Responses in the Lung1

    PubMed Central

    Ravindranath, Thyyar M.; Mong, Phyllus Y.; Ananthakrishnan, Radha; Li, Qing; Quadri, Nosirudeen; Schmidt, Ann Marie; Ramasamy, Ravichandran; Wang, Qin

    2011-01-01

    Exaggerated inflammatory responses and the resultant increases in alveolar-capillary permeability underlie the pathogenesis of acute lung injury during sepsis. This study examined the functions of aldose reductase (AR) in mediating acute lung inflammation. Transgenic mice expressing human AR (ARTg) were used to study the functions of AR since mice have low intrinsic AR activity. In a mild cecal ligation and puncture model, ARTg mice demonstrated an enhanced AR activity and a greater inflammatory response as evaluated by circulating cytokine levels, neutrophil accumulation in the lungs, and activation of Rho kinase in lung endothelial cells (ECs). Compared with WT lung cells, ARTg lung cells produced more IL-6 and showed augmented JNK activation in response to LPS stimulation ex vivo. In human neutrophils, AR activity was required for fMLP-included CD11b activation and up-regulation, respiratory burst, and shape changes. In human pulmonary microvascular ECs, AR activity was required for TNF-α-induced activation of the Rho kinase/MKK4/JNK pathway and IL-6 production, but not p38 activation or ICAM-1 expression. Importantly, AR activity in both human neutrophils and ECs was required for neutrophil adhesion to TNF-α-stimulated ECs. These data demonstrate a novel role for AR in regulating the signaling pathways leading to neutrophil-EC adhesion during acute lung inflammation. PMID:20007578

  7. Metabolism of the aldose reductase inhibitor ALO1567 in man.

    PubMed Central

    Park, Y H; Hudson, J E; Barker, R C; York, B M; Brazzell, R K

    1991-01-01

    1. The metabolism of the aldose reductase inhibitor, ALO1567, was studied in man. The major biotransformation pathway was aromatic hydroxylation followed by glucuronide conjugation. 2. Hydroxylation occurred at several positions on the fluorene ring. The major metabolite was identified as the 7-hydroxy analogue of ALO1567 and three minor metabolites were characterized as positional isomers of the 7-hydroxy metabolite. 3. Oxidative defluorination and metabolism on the hydantoin ring were also indicated as minor pathways. 4. The capacity of normal subjects to oxidize ALO1567 was indicated by the urinary ratio of the parent drug to the 7-hydroxy metabolite after daily oral administration of 100 mg and 200 mg of ALO1567. Most subjects having higher ALO1567 plasma concentrations showed higher ratios. PMID:1931471

  8. Diuretic and natriuretic effects of sorbinil, an aldose reductase inhibitor.

    PubMed

    Springate, J E; Feld, L G; Van Liew, J B; Fildes, R D; Acara, M A

    1991-04-01

    The renal effects of sorbinil, an aldose reductase inhibitor that interferes with the conversion of glucose to sorbitol, were studied in rats and rabbits before and after fluid deprivation. The intracellular osmolar solute, sorbitol, is found in increasing concentrations from cortex to medulla in the kidney and may be involved in the urinary concentrating mechanism. Oral administration of sorbinil in the rabbit resulted in significant increases in urine flow rate and sodium excretion with a tendency toward decreased urine osmolality and increased potassium excretion both before and after water deprivation. When fluid intake was controlled in the rat study, significant increases in urine flow rate and sodium and potassium excretion and a significant decrease in urine osmolality occurred only in response to fluid deprivation. Thus, sorbinil has diuretic and natriuretic properties and may prevent the normal concentration of urine in the antidiuretic animal.

  9. Effect of Polygonum hydropiper sulfated flavonoids on lens aldose reductase and related enzymes.

    PubMed

    Haraguchi, H; Ohmi, I; Sakai, S; Fukuda, A; Toihara, Y; Fujimoto, T; Okamura, N; Yagi, A

    1996-04-01

    The sulfated flavonoids in Polygonum hydropiper showed potent inhibiton against lens aldose reductase. Among these flavonoids isorhamnetin 3,7-disulfate (5) was most potent. Kinetic analysis showed that 5 exhibited noncompetitive inhibition against both dl-glyceraldehyde and NADPH.

  10. Studies on WF-3681, a novel aldose reductase inhibitor. I. Taxonomy, fermentation, isolation and characterization.

    PubMed

    Nishikawa, M; Tsurumi, Y; Namiki, T; Yoshida, K; Okuhara, M

    1987-10-01

    WF-3681 was isolated from a cultured filtrate of Chaetomella raphigera as a novel inhibitor of aldose reductase. It was extracted with ethyl acetate and then purified with silica gel chromatography. Its molecular formula was determined to be C13H12O5 by elemental analysis and high resolution electron impact mass spectrometry. IC50 of WF-3681 was 2.5 X 10(-7) M for partially purified aldose reductase of rabbit lens. PMID:3119547

  11. Aldose Reductase Mediates Endotoxin-Induced Production of Nitric oxide and Cytotoxicity in Murine Macrophages.

    PubMed Central

    Ramana, Kota V; Reddy, Aramati BM.; Tammali, Ravinder; Srivastava, Satish K.

    2007-01-01

    Aldose reductase (AR) is a ubiquitously expressed protein with pleiotrophic roles as an efficient catalyst for the reduction of toxic lipid aldehydes and mediator of hyperglycemia, cytokine and growth factor –induced redox sensitive signals that cause secondary diabetic complications. Although AR inhibition has been shown to be protective against oxidative stress signals, the role of AR in regulating nitric oxide (NO) synthesis and NO-mediated apoptosis has not been elucidated to date. We therefore investigated the role of AR in regulating lipopolysaccharide (LPS)-induced NO synthesis and apoptosis in RAW 264.7 macrophages. Inhibition or RNA interference ablation of AR suppressed LPS-stimulated production of NO and over-expression of iNOS mRNA. Inhibition or ablation of AR also prevented the LPS-induced apoptosis, cell cycle arrest, activation of caspase-3, p38-MAPK, JNK, NF-κB and AP1. In addition, AR inhibition prevented the LPS-induced down-regulation of Bcl-xl and up-regulation of Bax and Bak in macrophages. L-arginine increased and L-NAME decreased the severity of cell death caused by LPS and AR inhibitors prevented it. Furthermore, inhibition of AR prevents cell death caused by HNE and GS-HNE, but not GS-DHN. Our findings for the first time suggest that AR catalyzed lipid aldehyde-glutathione conjugates regulates the LPS-induced production of inflammatory marker NO and cytotoxicity in RAW 264.7 cells. Inhibition or ablation of AR activity may be potential therapeutic target in endotoximia and other inflammatory diseases. PMID:17382209

  12. Aldose Reductase Inhibition Alleviates Hyperglycemic Effects on Human Retinal Pigment Epithelial Cells

    PubMed Central

    Chang, Kun-Che; Snow, Anson; LaBarbera, Daniel V.; Petrash, J. Mark

    2014-01-01

    Chronic hyperglycemia is an important risk factor involved in the onset and progression of diabetic retinopathy (DR). Among other effectors, aldose reductase (AR) has been linked to the pathogenesis of this degenerative disease. The purpose of this study was to investigate whether the novel AR inhibitor, beta-glucogallin (BGG), can offer protection against various hyperglycemia-induced abnormalities in human adult retinal pigment epithelia (ARPE-19) cells. AR is an enzyme that contributes to cellular stress by production of reactive oxygen species (ROS) under high glucose conditions. A marked decrease in cell viability (from 100% to 78%) following long-term exposure (4 days) of RPE cells to high glucose (HG) was largely prevented by siRNA-mediated knockdown of AR gene expression (from 79% to 97%) or inhibition using sorbinil (from 66% to 86%). In HG, BGG decreased sorbitol accumulation (44%), ROS production (27%) as well as ER stress (22%). Additionally, we demonstrated that BGG prevented loss of mitochondrial membrane potential (MMP) under HG exposure. We also showed that AR inhibitor pretreatment reduced retinal microglia-induced apoptosis in APRE-19 cells. These results suggest that BGG may be useful as a therapeutic agent against retinal degeneration in the diabetic eye by preventing RPE cell death. PMID:25451566

  13. Aldose Reductase acts as a Selective Derepressor of PPARγ and Retinoic Acid Receptor

    PubMed Central

    Thiagarajan, Devi; Ananthakrishnan, Radha; Zhang, Jinghua; O’Shea, Karen M.; Quadri, Nosirudeen; Li, Qing; Sas, Kelli; Jing, Xiao; Rosario, Rosa; Pennathur, Subramaniam; Schmidt, Ann Marie; Ramasamy, Ravichandran

    2016-01-01

    Summary Histone deacetylase 3 (HDAC3), a chromatin modifying enzyme, requires association with the deacetylase containing domain (DAD) of the nuclear receptor co-repressors NCOR1 and SMRT for its stability and activity. Here we show that aldose reductase (AR), the rate-limiting enzyme of the polyol pathway, competes with HDAC3 to bind the NCOR1/SMRT DAD. Increased AR expression leads to HDAC3 degradation followed by increased PPARγ signaling resulting in lipid accumulation in the heart. AR also downregulates expression of nuclear corepressor complex cofactors including Gps2 and Tblr1, thus affecting activity of the nuclear corepressor complex itself. Though AR reduces HDAC3-corepressor complex formation, it specifically de-represses the retinoic acid receptor (RAR), but not other nuclear receptors such as the thyroid receptor (TR) and liver X receptor (LXR). In summary, this work defines a distinct role for AR in lipid and retinoid metabolism through HDAC3 regulation and consequent de-repression of PPARγ and RAR. PMID:27052179

  14. Anti-neuroinflammatory efficacy of the aldose reductase inhibitor FMHM via phospholipase C/protein kinase C-dependent NF-κB and MAPK pathways

    SciTech Connect

    Zeng, Ke-Wu; Li, Jun; Dong, Xin; Wang, Ying-Hong; Ma, Zhi-Zhong; Jiang, Yong; Jin, Hong-Wei; Tu, Peng-Fei

    2013-11-15

    Aldose reductase (AR) has a key role in several inflammatory diseases: diabetes, cancer and cardiovascular diseases. Therefore, AR inhibition seems to be a useful strategy for anti-inflammation therapy. In the central nervous system (CNS), microglial over-activation is considered to be a central event in neuroinflammation. However, the effects of AR inhibition in CNS inflammation and its underlying mechanism of action remain unknown. In the present study, we found that FMHM (a naturally derived AR inhibitor from the roots of Polygala tricornis Gagnep.) showed potent anti-neuroinflammatory effects in vivo and in vitro by inhibiting microglial activation and expression of inflammatory mediators. Mechanistic studies showed that FMHM suppressed the activity of AR-dependent phospholipase C/protein kinase C signaling, which further resulted in downstream inactivation of the IκB kinase/IκB/nuclear factor-kappa B (NF-κB) inflammatory pathway. Therefore, AR inhibition-dependent NF-κB inactivation negatively regulated the transcription and expression of various inflammatory genes. AR inhibition by FMHM exerted neuroprotective effects in lipopolysaccharide-induced neuron–microglia co-cultures. These findings suggested that AR is a potential target for neuroinflammation inhibition and that FMHM could be an effective agent for treating or preventing neuroinflammatory diseases. - Highlights: • FMHM is a natural-derived aldose reductase (AR) inhibitor. • FMHM inhibits various neuroinflammatory mediator productions in vitro and in vivo. • FMHM inhibits neuroinflammation via aldose reductase/PLC/PKC-dependent NF-κB pathway. • FMHM inhibits neuroinflammation via aldose reductase/PLC/PKC-dependent MAPK pathway. • FMHM protects neurons against inflammatory injury in microglia-neuron co-cultures.

  15. Inhibition of Aldose Reductase by Gentiana lutea Extracts

    PubMed Central

    Akileshwari, Chandrasekhar; Muthenna, Puppala; Nastasijević, Branislav; Joksić, Gordana; Petrash, J. Mark; Reddy, Geereddy Bhanuprakash

    2012-01-01

    Accumulation of intracellular sorbitol due to increased aldose reductase (ALR2) activity has been implicated in the development of various secondary complications of diabetes. Thus, ALR2 inhibition could be an effective strategy in the prevention or delay of certain diabetic complications. Gentiana lutea grows naturally in the central and southern areas of Europe. Its roots are commonly consumed as a beverage in some European countries and are also known to have medicinal properties. The water, ethanol, methanol, and ether extracts of the roots of G. lutea were subjected to in vitro bioassay to evaluate their inhibitory activity on the ALR2. While the ether and methanol extracts showed greater inhibitory activities against both rat lens and human ALR2, the water and ethanol extracts showed moderate inhibitory activities. Moreover, the ether and methanol extracts of G. lutea roots significantly and dose-dependently inhibited sorbitol accumulation in human erythrocytes under high glucose conditions. Molecular docking studies with the constituents commonly present in the roots of G. lutea indicate that a secoiridoid glycoside, amarogentin, may be a potential inhibitor of ALR2. This is the first paper that shows G. lutea extracts exhibit inhibitory activity towards ALR2 and these results suggest that Gentiana or its constituents might be useful to prevent or treat diabetic complications. PMID:22844269

  16. Inhibition of aldose reductase by Gentiana lutea extracts.

    PubMed

    Akileshwari, Chandrasekhar; Muthenna, Puppala; Nastasijević, Branislav; Joksić, Gordana; Petrash, J Mark; Reddy, Geereddy Bhanuprakash

    2012-01-01

    Accumulation of intracellular sorbitol due to increased aldose reductase (ALR2) activity has been implicated in the development of various secondary complications of diabetes. Thus, ALR2 inhibition could be an effective strategy in the prevention or delay of certain diabetic complications. Gentiana lutea grows naturally in the central and southern areas of Europe. Its roots are commonly consumed as a beverage in some European countries and are also known to have medicinal properties. The water, ethanol, methanol, and ether extracts of the roots of G. lutea were subjected to in vitro bioassay to evaluate their inhibitory activity on the ALR2. While the ether and methanol extracts showed greater inhibitory activities against both rat lens and human ALR2, the water and ethanol extracts showed moderate inhibitory activities. Moreover, the ether and methanol extracts of G. lutea roots significantly and dose-dependently inhibited sorbitol accumulation in human erythrocytes under high glucose conditions. Molecular docking studies with the constituents commonly present in the roots of G. lutea indicate that a secoiridoid glycoside, amarogentin, may be a potential inhibitor of ALR2. This is the first paper that shows G. lutea extracts exhibit inhibitory activity towards ALR2 and these results suggest that Gentiana or its constituents might be useful to prevent or treat diabetic complications.

  17. Overexpression of Aldose Reductase Render Mouse Hepatocytes More Sensitive to Acetaminophen Induced Oxidative Stress and Cell Death.

    PubMed

    Ahmed, Munzir M E; Al-Obosi, J A S; Osman, H M; Shayoub, M E

    2016-04-01

    Acetaminophen (APAP) a commonly used drug for decrease the fever and pain but is capable to induced hepatotoxicity at over dose. This study was carried out to investigate the effect of APAP on the expression of anti-apoptotic and antioxidative defense genes, and whether aldose reductase over-expressing plasmid capable to protect against APAP-induced oxidative stress and cell death. APAP treatment induced oxidative stress and hepatotoxicity, and significantly increased aldose reductase mRNA and protein expression in mouse hepatocyte (AML-12). Unexpectedly, AML-12 cells over-expressing aldose reductase augmented APAP-induced reduction in cell viability, reactive oxygen species (ROS) production, glutathione (GSH) depletion and glutathione S-transferase A2 expression. Moreover, over-expression of aldose reductase potentiated APAP induced reduction on proliferating cell nuclear antigen, B cell lymphoma-extra large (bcl-xL), catalase, glutathione peroxidase-1 (GPx-1) and abolished APAP-induced B-cell lymphoma 2 (bcl-2) inductions. Further, over-expression of aldose reductase significantly abolished AMP activated protein kinase (AMPK) activity in APAP-treated cells and induced p53 expression. This results demonstrate that APAP induced toxicity in AML-12, increased aldose reductase expression, and over-expression of aldose reductase render this cell more susceptible to APAP induced oxidative stress and cell death, this probably due to inhibition AMPK or bcl-2 activity, or may due to competition between aldose reductase and glutathione reductase for NADPH. PMID:27069324

  18. Identification of Novel Aldose Reductase Inhibitors from Spices: A Molecular Docking and Simulation Study.

    PubMed

    Antony, Priya; Vijayan, Ranjit

    2015-01-01

    Hyperglycemia in diabetic patients results in a diverse range of complications such as diabetic retinopathy, neuropathy, nephropathy and cardiovascular diseases. The role of aldose reductase (AR), the key enzyme in the polyol pathway, in these complications is well established. Due to notable side-effects of several drugs, phytochemicals as an alternative has gained considerable importance for the treatment of several ailments. In order to evaluate the inhibitory effects of dietary spices on AR, a collection of phytochemicals were identified from Zingiber officinale (ginger), Curcuma longa (turmeric) Allium sativum (garlic) and Trigonella foenum graecum (fenugreek). Molecular docking was performed for lead identification and molecular dynamics simulations were performed to study the dynamic behaviour of these protein-ligand interactions. Gingerenones A, B and C, lariciresinol, quercetin and calebin A from these spices exhibited high docking score, binding affinity and sustained protein-ligand interactions. Rescoring of protein ligand interactions at the end of MD simulations produced binding scores that were better than the initially docked conformations. Docking results, ligand interactions and ADMET properties of these molecules were significantly better than commercially available AR inhibitors like epalrestat, sorbinil and ranirestat. Thus, these natural molecules could be potent AR inhibitors. PMID:26384019

  19. Identification of Novel Aldose Reductase Inhibitors from Spices: A Molecular Docking and Simulation Study

    PubMed Central

    Antony, Priya; Vijayan, Ranjit

    2015-01-01

    Hyperglycemia in diabetic patients results in a diverse range of complications such as diabetic retinopathy, neuropathy, nephropathy and cardiovascular diseases. The role of aldose reductase (AR), the key enzyme in the polyol pathway, in these complications is well established. Due to notable side-effects of several drugs, phytochemicals as an alternative has gained considerable importance for the treatment of several ailments. In order to evaluate the inhibitory effects of dietary spices on AR, a collection of phytochemicals were identified from Zingiber officinale (ginger), Curcuma longa (turmeric) Allium sativum (garlic) and Trigonella foenum graecum (fenugreek). Molecular docking was performed for lead identification and molecular dynamics simulations were performed to study the dynamic behaviour of these protein-ligand interactions. Gingerenones A, B and C, lariciresinol, quercetin and calebin A from these spices exhibited high docking score, binding affinity and sustained protein-ligand interactions. Rescoring of protein ligand interactions at the end of MD simulations produced binding scores that were better than the initially docked conformations. Docking results, ligand interactions and ADMET properties of these molecules were significantly better than commercially available AR inhibitors like epalrestat, sorbinil and ranirestat. Thus, these natural molecules could be potent AR inhibitors. PMID:26384019

  20. Identification of Novel Aldose Reductase Inhibitors from Spices: A Molecular Docking and Simulation Study.

    PubMed

    Antony, Priya; Vijayan, Ranjit

    2015-01-01

    Hyperglycemia in diabetic patients results in a diverse range of complications such as diabetic retinopathy, neuropathy, nephropathy and cardiovascular diseases. The role of aldose reductase (AR), the key enzyme in the polyol pathway, in these complications is well established. Due to notable side-effects of several drugs, phytochemicals as an alternative has gained considerable importance for the treatment of several ailments. In order to evaluate the inhibitory effects of dietary spices on AR, a collection of phytochemicals were identified from Zingiber officinale (ginger), Curcuma longa (turmeric) Allium sativum (garlic) and Trigonella foenum graecum (fenugreek). Molecular docking was performed for lead identification and molecular dynamics simulations were performed to study the dynamic behaviour of these protein-ligand interactions. Gingerenones A, B and C, lariciresinol, quercetin and calebin A from these spices exhibited high docking score, binding affinity and sustained protein-ligand interactions. Rescoring of protein ligand interactions at the end of MD simulations produced binding scores that were better than the initially docked conformations. Docking results, ligand interactions and ADMET properties of these molecules were significantly better than commercially available AR inhibitors like epalrestat, sorbinil and ranirestat. Thus, these natural molecules could be potent AR inhibitors.

  1. Bioactive fraction of Saraca indica prevents diabetes induced cataractogenesis: An aldose reductase inhibitory activity

    PubMed Central

    Somani, Gauresh; Sathaye, Sadhana

    2015-01-01

    Background: The present study was designed to investigate the effect of Saraca indica (SI) flowers extract and different bioactive fraction on in vitro aldose reductase (AR) inhibitory activity, high glucose-induced cataract in goat lens and in vivo streptozotocin (STZ; 45 mg/kg, i.p) induced cataract in rats. Methods: Extract of flowers of SI tested for inhibition against rat lens AR. Furthermore, bioactive fraction was investigated against high glucose-induced opacification of the lens in vitro lens culture and STZ induced diabetic cataract in rats. Identification of the bioactive component was attempted through high-performance thin-layer chromatography, high-performance liquid chromatography and liquid chromatography-mass spectrometry analysis. Results: Ethyl acetate fraction of S. indica (EASI) produced maximum inhibition that may be due to high phenolic content. Goat lenses in media containing glucose developed a distinctly opaque ring in 72 h and treatment with EASI fraction lowered lens opacity in 72 h. Prolonged treatment with EASI to STZ-induced diabetic rats inhibited the AR activity and delayed cataract progression in a dose dependent manner. Conclusion: Ethyl acetate fraction of S. indica fraction has potential to inhibit rat lens AR enzyme and prevent cataractogenesis not only in goat lens model (in vitro), but also in STZ induced diabetic rats (in vivo). This study is suggestive of the anticataract activity of EASI fraction that could be attributed to the phytoconstituents present in the same. PMID:25709218

  2. Advance in dietary polyphenols as aldose reductases inhibitors: structure-activity relationship aspect.

    PubMed

    Xiao, Jianbo; Ni, Xiaoling; Kai, Guoyin; Chen, Xiaoqing

    2015-01-01

    The dietary polyphenols as aldose reductases inhibitors (ARIs) have attracted great interest among researchers. The aim of this review is to give an overview of the research reports on the structure-activity relationship of dietary polyphenols inhibiting aldose reductases (AR). The molecular structures influence the inhibition of the following: (1) The methylation and methoxylation of the hydroxyl group at C3, C3', and C4' of flavonoids decreased or little affected the inhibitory potency. However, the methylation and methoxylation of the hydroxyl group at C5, C6, and C8 significantly enhanced the inhibition. Moreover, the methylation and methoxylation of C7-OH influence the inhibitory activity depending on the substitutes on rings A and B of flavonoids. (2) The glycosylation on 3-OH of flavonoids significantly increased or little affected the inhibition. However, the glycosylation on 7-OH and 4'-OH of flavonoids significantly decreased the inhibition. (3) The hydroxylation on A-ring of flavones and isoflavones, especially at positions 5 and 7, significantly improved the inhibition and the hydroxylation on C3' and C4' of B-ring of flavonoids remarkably enhanced the inhibition; however, the hydroxylation on the ring C of flavones significantly weakened the inhibition. (4) The hydrogenation of the C2=C3 double bond of flavones reduced the inhibition. (5) The hydrogenation of α=β double bond of stilbenes hardly affected the inhibition and the hydroxylation on C3' of stilbenes decreased the inhibition. Moreover, the methylation of the hydroxyl group of stilbenes obviously reduced the activity. (6) The hydroxylation on C4 of chalcone significantly increased the inhibition and the methylation on C4 of chalcone remarkably weakened the inhibition.

  3. Cuminaldehyde: Aldose Reductase and alpha-Glucosidase Inhibitor Derived from Cuminum cyminum L. Seeds.

    PubMed

    Lee, Hoi-Seon

    2005-04-01

    The inhibitory activity of Cuminum cyminum seed-isolated component was evaluated against lens aldose reductase and alpha-glucosidase isolated from Sprague-Dawley male rats and compared to that of 11 commercially available components derived from C. cyminum seed oil, as well as quercitrin as an aldose reductase inhibitor and acarbose as an alpha-glucosidase inhibitor. The biologically active constituent of C. cyminum seed oil was characterized as cuminaldehyde by various spectral analyses. The IC(50) value of cuminaldehyde is 0.00085 mg/mL against aldose reductase and 0.5 mg/mL against alpha-glucosidase, respectively. Cuminaldehyde was about 1.8 and 1.6 times less in inhibitory activity than acarbose and quercitin, respectively. Nonetheless, cuminaldehyde may be useful as a lead compound and a new agent for antidiabetic therapeutics.

  4. [Progress in research of aldose reductase inhibitors in traditional medicinal herbs].

    PubMed

    Feng, Chang-Gen; Zhang, Lin-Xia; Liu, Xia

    2005-10-01

    The traditional medicinal herbs are natural product, and have no obviously toxic action and side effect, and their resources are extensive. The adverse effects produced by aldose reductase inhibitors in traditional medicinal herbs are less than those from chemical synthesis and micro-organism, they can effectively prevent and delay diabetic complication, such as diabetic nephropathy, vasculopathy, retinopathy, peripheral neuropathy, and so on. They will have a wonderful respect. Flavonoid compounds and their derivates from traditional medicinal herbs are active inhibitors to aldose reductase, such as quercetin, silymarin, puerarin, baicalim, berberine and so on. In addition, some compound preparations show more strongly activity in inhibiting aldose reductase and degrading sorbitol contents, such as Shendan in traditional medicinal herbs being active inhibitors and Jianyi capsule, Jinmaitong composita, Liuwei Di-huang pill, et al. The progresses definite functions of treating diabetes complications have been reviewed.

  5. Isolation, modification, and aldose reductase inhibitory activity of rosmarinic acid derivatives from the roots of Salvia grandifolia.

    PubMed

    Kang, Jie; Tang, Yanbo; Liu, Quan; Guo, Nan; Zhang, Jian; Xiao, Zhiyan; Chen, Ruoyun; Shen, Zhufang

    2016-07-01

    To find aldose reductase inhibitors, two previously unreported compounds, grandifolias H and I, and five known compounds, including rosmarinic acid and rosmarinic acid derivatives, were isolated from the roots of Salvia grandifolia. A series of rosmarinic acid derivatives was obtained from rosmarinic acid using simple synthetic methods. The aldose reductase inhibitory activity of the isolated and synthesized compounds was assessed. Seven of the tested compounds showed moderate aldose reductase inhibition (IC50=0.06-0.30μM). The structure-activity relationship of aldose reductase inhibitory activity of rosmarinic acid derivatives was discussed for the first time. This study provided useful information that will facilitate the development of aldose reductase inhibitors. PMID:27233987

  6. Aldose reductase regulates acrolein-induced cytotoxicity in human small airway epithelial cells.

    PubMed

    Yadav, Umesh C S; Ramana, K V; Srivastava, Satish K

    2013-12-01

    Aldose reductase (AR), a glucose-metabolizing enzyme, reduces lipid aldehydes and their glutathione conjugates with more than 1000-fold efficiency (Km aldehydes 5-30 µM) relative to glucose. Acrolein, a major endogenous lipid peroxidation product as well as a component of environmental pollutants and cigarette smoke, is known to be involved in various pathologies including atherosclerosis, airway inflammation, COPD, and age-related disorders, but the mechanism of acrolein-induced cytotoxicity is not clearly understood. We have investigated the role of AR in acrolein-induced cytotoxicity in primary human small airway epithelial cells (SAECs). Exposure of SAECs to varying concentrations of acrolein caused cell death in a concentration- and time-dependent manner. AR inhibition by fidarestat prevented the low-dose (5-10 µM) but not the high-dose (>10 µM) acrolein-induced SAEC death. AR inhibition protected SAECs from low-dose (5 µM) acrolein-induced cellular reactive oxygen species (ROS). Inhibition of acrolein-induced apoptosis by fidarestat was confirmed by decreased condensation of nuclear chromatin, DNA fragmentation, comet tail moment, and annexin V fluorescence. Further, fidarestat inhibited acrolein-induced translocation of the proapoptotic proteins Bax and Bad from the cytosol to the mitochondria and that of Bcl2 and BclXL from the mitochondria to the cytosol. Acrolein-induced cytochrome c release from mitochondria was also prevented by AR inhibition. The mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinases 1 and 2, stress-activated protein kinase/c-Jun NH2-terminal kinase, and p38MAPK, and c-Jun were transiently activated in airway epithelial cells by acrolein in a concentration- and time-dependent fashion, which was significantly prevented by AR inhibition. These results suggest that AR inhibitors could prevent acrolein-induced cytotoxicity in the lung epithelial cells.

  7. Affinity purifications of aldose reductase and xylitol dehydrogenase from the xylose-fermenting yeast Pachysolen tannophilus

    SciTech Connect

    Bolen, P.L.; Roth, K.A.; Freer, S.N.

    1986-10-01

    Although xylose is a major product of hydrolysis of lignocellulosic materials, few yeasts are able to convert it to ethanol. In Pachysolen tannophilus, one of the few xylose-fermenting yeasts found, aldose reductase and xylitol dehydrogenase were found to be key enzymes in the metabolic pathway for xylose fermentation. This paper presents a method for the rapid and simultaneous purification of both aldose reductase and xylitol dehydrogenase from P. tannophilus. Preliminary studies indicate that this method may be easily adapted to purify similar enzymes from other xylose-fermenting yeasts.

  8. Nrf2 regulates curcumin-induced aldose reductase expression indirectly via nuclear factor-kappaB.

    PubMed

    Kang, Eun Sil; Kim, Gil Hyeong; Kim, Hyo Jung; Woo, Im Sun; Ham, Sun Ah; Jin, Hana; Kim, Min Young; Kim, Hye Jung; Lee, Jae Heun; Chang, Ki Churl; Seo, Han Geuk; Hwang, Jin-Yong

    2008-07-01

    The osmotic response element (ORE) differs from the nuclear factor-kappaB (NF-kappaB) binding sequence by a single base pair; therefore, we investigated the involvement of NF-kappaB in the induction of aldose reductase (AR) by curcumin. Curcumin, an herb-derived polyphenolic compound, elicited an increase in the expression and promoter activity of the AR gene in a nuclear factor-erythroid 2-related factor 2 (Nrf2)-dependent manner. Small interfering RNA (siRNA) against p65 or BAY11-7082, an inhibitor of NF-kappaB, significantly suppressed the curcumin and/or Nrf2-induced increase in expression levels and promoter activity of the AR gene. BAY11-7082 or siRNA against p65 also attenuated the curcumin-induced increase in the promoter activity of the wild type AR-ORE(wt) gene, but not that of the mutated AR-ORE(mt), indicating that the ORE is essential for the response to NF-kappaB. The expression of p65, the promoter activity and DNA binding activity of NF-kappaB were enhanced in the presence of curcumin in cells that were transfected with Nrf2 compared to those treated with curcumin alone. Cells that had been preincubated with curcumin demonstrated resistance to reactive oxygen species-induced cell damage through the suppressive effects in the generation of reactive aldehydes. These effects were significantly attenuated in the presence of BAY11-7082, indicating the involvement of NF-kappaB in the cellular response of AR to oxidative stress and toxic aldehydes.

  9. Role of Aldose Reductase in the Metabolism and Detoxification of Carnosine-Acrolein Conjugates*

    PubMed Central

    Baba, Shahid P.; Hoetker, Joseph David; Merchant, Michael; Klein, Jon B.; Cai, Jian; Barski, Oleg A.; Conklin, Daniel J.; Bhatnagar, Aruni

    2013-01-01

    Oxidation of unsaturated lipids generates reactive aldehydes that accumulate in tissues during inflammation, ischemia, or aging. These aldehydes form covalent adducts with histidine-containing dipeptides such as carnosine and anserine, which are present in high concentration in skeletal muscle, heart, and brain. The metabolic pathways involved in the detoxification and elimination of these conjugates are, however, poorly defined, and their significance in regulating oxidative stress is unclear. Here we report that conjugates of carnosine with aldehydes such as acrolein are produced during normal metabolism and excreted in the urine of mice and adult human non-smokers as carnosine-propanols. Our studies show that the reduction of carnosine-propanals is catalyzed by the enzyme aldose reductase (AR). Carnosine-propanals were converted to carnosine-propanols in the lysates of heart, skeletal muscle, and brain tissue from wild-type (WT) but not AR-null mice. In comparison with WT mice, the urinary excretion of carnosine-propanols was decreased in AR-null mice. Carnosine-propanals formed covalent adducts with nucleophilic amino acids leading to the generation of carnosinylated proteins. Deletion of AR increased the abundance of proteins bound to carnosine in skeletal muscle, brain, and heart of aged mice and promoted the accumulation of carnosinylated proteins in hearts subjected to global ischemia ex vivo. Perfusion with carnosine promoted post-ischemic functional recovery in WT but not in AR-null mouse hearts. Collectively, these findings reveal a previously unknown metabolic pathway for the removal of carnosine-propanal conjugates and suggest a new role of AR as a critical regulator of protein carnosinylation and carnosine-mediated tissue protection. PMID:23928303

  10. High-resolution neutron protein crystallography with radically small crystal volumes: Application of perdeuteration to human aldose reductase

    SciTech Connect

    Hazemann, I.; Dauvergne, M. T.; Blakeley, M. P.; Meilleur, Flora; Haertlein, M.; Van Dorsselaer, A.; Mitschler, A.; Myles, Dean A A; Podjarny, A.

    2005-08-01

    Neutron diffraction data have been collected to 2.2 {angstrom} resolution from a small (0.15 mm{sup 3}) crystal of perdeuterated human aldose reductase (h-AR; MW = 36 kDa) in order to help to determine the protonation state of the enzyme. h-AR belongs to the aldo-keto reductase family and is implicated in diabetic complications. Its ternary complexes (h-AR-coenzyme NADPH-selected inhibitor) provide a good model to study both the enzymatic mechanism and inhibition. Here, the successful production of fully deuterated human aldose reductase [h-AR(D)], subsequent crystallization of the ternary complex h-AR(D)-NADPH-IDD594 and neutron Laue data collection at the LADI instrument at ILL using a crystal volume of just 0.15 mm{sup 3} are reported. Neutron data were recorded to 2 {angstrom} resolution, with subsequent data analysis using data to 2.2 {angstrom}. This is the first fully deuterated enzyme of this size (36 kDa) to be solved by neutron diffraction and represents a milestone in the field, as the crystal volume is at least one order of magnitude smaller than those usually required for other high-resolution neutron structures determined to date. This illustrates the significant increase in the signal-to-noise ratio of data collected from perdeuterated crystals and demonstrates that good-quality neutron data can now be collected from more typical protein crystal volumes. Indeed, the signal-to-noise ratio is then dominated by other sources of instrument background, the nature of which is under investigation. This is important for the design of future instruments, which should take maximum advantage of the reduction in the intrinsic diffraction pattern background from fully deuterated samples.

  11. Diallyl sulfide protects against N-nitrosodiethylamine-induced liver tumorigenesis: Role of aldose reductase

    PubMed Central

    Ibrahim, Safinaz S; Nassar, Noha N

    2008-01-01

    AIM: To evaluate the protective effect of diallyl sulfide (DAS) against N-nitrosodiethylamine (NDEA)-induced liver carcinogenesis. METHODS: Male Wistar rats received either NDEA or NDEA together with DAS as protection. Liver energy metabolism was assessed in terms of lactate, pyruvate, lactate/pyruvate, ATP levels, lactate dehydrogenase (LDH) and glucose-6-phosphate dehydrogenase (G6PD) activities. In addition, membrane disintegration of the liver cells was evaluated by measuring lipid-peroxidation products, measured as malondialdehyde (MDA); nitric oxide (NO) levels; glucose-6-phosphatase (G6Pase), catalase (CAT) and superoxide dismutase (SOD) activities. Liver DNA level, glutathione-S-transferase (GST) and cytochrome c oxidase activities were used as DNA fragmentation indices. Aldose reductase (AR) activity was measured as an index for cancer cells resistant to chemotherapy and histopathological examination was performed on liver sections from different groups. RESULTS: NDEA significantly disturbed liver functions and most of the aforementioned indices. Treatment with DAS significantly restored liver functions and hepatocellular integrity; improved parameters of energy metabolism and suppressed free-radical generation. CONCLUSION: We provide evidence that DAS exerts a protective role on liver functions and tissue integrity in face of enhanced tumorigenesis caused by NDEA, as well as improving cancer-cell sensitivity to chemotherapy. This is mediated through combating oxidative stress of free radicals, improving the energy metabolic state of the cell, and enhancing the activity of G6Pase, GST and AR enzymes. PMID:18985804

  12. In vitro antidiabetic effects of selected fruits and vegetables against glycosidase and aldose reductase.

    PubMed

    Wu, Tong; Luo, Jiaqiang; Xu, Baojun

    2015-11-01

    In vitro antidiabetic effect of fruits and vegetables with reports as folk remedies were investigated. The antidiabetic effects were evaluated by comparing the inhibitory properties of α-glycosidase, aldose reductase, and antioxidant activity. The results indicated that lychee extract exhibited the best dose-dependent inhibitory activity against α-glycosidase with IC 50 of 10.4 mg/mL, and lemon peel extract exhibited aldose reductase inhibitory potential with IC 50 value at 3.63 mg/mL. Besides, the result also showed that the inhibitory effects of blueberry and plum against α-glycosidase were strong among the fruits samples. Bitter gourd and eggplant demonstrated significant inhibitory potential against aldose reductase, with IC 50 values at 8.55 mg/mL and 8.06 mg/mL, respectively. The result from correlation analysis part showed that the antioxidant activities of selected fruits and vegetables were found related to their health beneficial effects, as there was positive correlations between total flavonoids content (TFC) and aldose reductase inhibitory activity (r (2) = 0.556). PMID:26788291

  13. Aldose reductases influence prostaglandin F2α levels and adipocyte differentiation in male mouse and human species.

    PubMed

    Pastel, Emilie; Pointud, Jean-Christophe; Loubeau, Gaëlle; Dani, Christian; Slim, Karem; Martin, Gwenaëlle; Volat, Fanny; Sahut-Barnola, Isabelle; Val, Pierre; Martinez, Antoine; Lefrançois-Martinez, Anne-Marie

    2015-05-01

    Aldose reductases (AKR1B) are widely expressed oxidoreductases whose physiological function remains elusive. Some isoforms are genuine prostaglandin F2α (PGF2α) synthases, suggesting they might influence adipose homeostasis because PGF2α inhibits adipogenesis. This was shown by Akr1b7 gene ablation in the mouse, which resulted in increased adiposity related to a lower PGF2α content in fat. Yet humans have no ortholog gene for Akr1b7, so the role of aldose reductases in human adipose homeostasis remains to be explored. We analyzed expression of genes encoding human and mouse aldose reductase isoforms in adipose tissues and differentiating adipocytes to assess conserved mechanisms regulating PGF2α synthesis and adipogenesis. The Akr1b3 gene encoded the most abundant isoform in mouse adipose tissue, whereas Akr1b7 encoded the only isoform enriched in the stromal vascular fraction. Most mouse aldose reductase gene expression peaked in early adipogenesis of 3T3-L1 cells and diminished with differentiation. In contrast with its mouse ortholog Akr1b3, AKR1B1 expression increased throughout differentiation of human multipotent adipose-derived stem cells, paralleling PGF2α release, whereas PGF2α receptor (FP) levels collapsed in early differentiation. Pharmacological inhibition of aldose reductase using Statil altered PGF2α production and enhanced human multipotent adipose-derived stem adipocyte differentiation. As expected, the adipogenic effects of Statil were counteracted by an FP agonist (cloprostenol). Thus, in both species aldose reductase-dependent PGF2α production could be important in early differentiation to restrict adipogenesis. PGF2α antiadipogenic signaling could then be toned down through the FP receptor or aldose reductases down-regulation in human and mouse cells, respectively. Our data suggest that aldose reductase inhibitors could have obesogenic potential.

  14. Structure of constituents isolated from the flower buds of Cananga odorata and their inhibitory effects on aldose reductase.

    PubMed

    Matsumoto, Takahiro; Nakamura, Seikou; Fujimoto, Katsuyoshi; Ohta, Tomoe; Ogawa, Keiko; Yoshikawa, Masayuki; Matsuda, Hisashi

    2014-10-01

    Three new terpenoid derivatives, canangaterpenes IV-VI, were isolated from the flower buds of Cananga odorata, cultivated in Thailand, together with eight known flavonoids. The chemical structures of the new compounds were elucidated on the basis of chemical and physicochemical evidence. The inhibitory effects of the isolated compounds on aldose reductase were also investigated. Several terpenoid derivatives and flavonoids were shown to inhibit aldose reductase. PMID:24816646

  15. Effects of aldose reductase inhibitor treatment in diabetic polyneuropathy - a clinical and neurophysiological study.

    PubMed Central

    Fagius, J; Jameson, S

    1981-01-01

    The efficacy of treatment with an aldose reductase inhibitor (1,3-dioxo-1 H-benz-de-isoquinoline-2(3H)-acetic acid, AY-22,284, Alrestatin) on peripheral nerve function in diabetic polyneuropathy was assessed. Thirty patients with long-standing diabetes and slight to moderate neuropathy participated in the double-blind placebo trial. Clinical examination, sensory threshold determinations for vibratory, tactile and thermal stimuli, conduction velocity measurements and studies of automatic function were performed to evaluate the treatment. Significant differences favouring Alrestatin over placebo were found for many of the measured variables, whereas no changes occurred on placebo. The apparent improvement of neuropathy occurred despite persisting hyperglycaemia. The results indicate that aldose reductase inhibitor treatment may be of value in diabetic polyneuropathy, and provide support for the sorbitol pathway hypothesis of diabetic polyneuropathy. PMID:6801211

  16. Melatonin Reduces Cataract Formation and Aldose Reductase Activity in Lenses of Streptozotocin-induced Diabetic Rat

    PubMed Central

    Khorsand, Marjan; Akmali, Masoumeh; Sharzad, Sahab; Beheshtitabar, Mojtaba

    2016-01-01

    Background: The relationship between the high activity of aldose reductase (AR) and diabetic cataract formation has been previously investigated. The purpose of the present study was to determine the preventing effect of melatonin on streptozotocin (STZ)-induced diabetic cataract in rats. Methods: 34 adult healthy male Sprague-Dawely rats were divided into four groups. Diabetic control and diabetic+melatonin received a single dose of STZ (50 mg/kg, intraperitoneally), whereas the normal control and normal+melatonin received vehicle. The melatonin groups were gavaged with melatonin (5 mg/kg) daily for a period of 8 weeks, whereas the rats in the normal control and diabetic control groups received only the vehicle. The rats’ eyes were examined every week and cataract formation scores (0-4) were determined by slit-lamp microscope. At the end of the eighth week, the rats were sacrificed and markers of the polyol pathway and antioxidative (Glutathione, GSH) in their lens were determined. The levels of blood glucose, HbA1c and plasma malondialdhyde (MDA), as a marker of lipid peroxidation, were also measured. Results: Melatonin prevented STZ-induced hyperglycemia by decreased blood glucose and HbA1c levels. Slit lamp examination indicated that melatonin delayed cataract progression in diabetic rats. The results revealed that melatonin feeding increased the GSH levels, decreased the activities of AR and sorbitol dehydrogenase (SDH) and sorbitol formation in catractous lenses as well as plasma MDA content. Conclusion: In summary, for the first time we demonstrated that melatonin delayed the formation and progression of cataract in diabetic rat lenses. PMID:27365552

  17. Inhibitory effects of Colocasia esculenta (L.) Schott constituents on aldose reductase.

    PubMed

    Li, Hong Mei; Hwang, Seung Hwan; Kang, Beom Goo; Hong, Jae Seung; Lim, Soon Sung

    2014-01-01

    The goal of this study was to determine the rat lens aldose reductase-inhibitory effects of 95% ethanol extracts from the leaves of C. esculenta and, its organic solvent soluble fractions, including the dichloromethane (CH2Cl2), ethyl acetate (EtOAc), n-butanol (BuOH) and water (H2O) layers, using dl-glyceraldehyde as a substrate. Ten compounds, namely tryptophan (1), orientin (2), isoorientin (3), vitexin (4), isovitexin (5), luteolin-7-O-glucoside (6), luteolin-7-O-rutinoside (7), rosmarinic acid (8), 1-O-feruloyl-d-glucoside (9) and 1-O-caffeoyl-d-glucoside (10) were isolated from the EtOAc and BuOH fractions of C. esculenta. The structures of compounds 1-10 were elucidated by spectroscopic methods and comparison with previous reports. All the isolates were subjected to an in vitro bioassay to evaluate their inhibitory activity against rat lens aldose reductase. Among tested compounds, compounds 2 and 3 significantly inhibited rat lens aldose reductase, with IC50 values of 1.65 and 1.92 μM, respectively. Notably, the inhibitory activity of orientin was 3.9 times greater than that of the positive control, quercetin (4.12 μM). However, the isolated compounds showed only moderate ABTS+ [2,29-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)] activity. These results suggest that flavonoid derivatives from Colocasia esculenta (L.) Schott represent potential compounds for the prevention and/or treatment of diabetic complications.

  18. Inhibitory effects of Colocasia esculenta (L.) Schott constituents on aldose reductase.

    PubMed

    Li, Hong Mei; Hwang, Seung Hwan; Kang, Beom Goo; Hong, Jae Seung; Lim, Soon Sung

    2014-01-01

    The goal of this study was to determine the rat lens aldose reductase-inhibitory effects of 95% ethanol extracts from the leaves of C. esculenta and, its organic solvent soluble fractions, including the dichloromethane (CH2Cl2), ethyl acetate (EtOAc), n-butanol (BuOH) and water (H2O) layers, using dl-glyceraldehyde as a substrate. Ten compounds, namely tryptophan (1), orientin (2), isoorientin (3), vitexin (4), isovitexin (5), luteolin-7-O-glucoside (6), luteolin-7-O-rutinoside (7), rosmarinic acid (8), 1-O-feruloyl-d-glucoside (9) and 1-O-caffeoyl-d-glucoside (10) were isolated from the EtOAc and BuOH fractions of C. esculenta. The structures of compounds 1-10 were elucidated by spectroscopic methods and comparison with previous reports. All the isolates were subjected to an in vitro bioassay to evaluate their inhibitory activity against rat lens aldose reductase. Among tested compounds, compounds 2 and 3 significantly inhibited rat lens aldose reductase, with IC50 values of 1.65 and 1.92 μM, respectively. Notably, the inhibitory activity of orientin was 3.9 times greater than that of the positive control, quercetin (4.12 μM). However, the isolated compounds showed only moderate ABTS+ [2,29-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)] activity. These results suggest that flavonoid derivatives from Colocasia esculenta (L.) Schott represent potential compounds for the prevention and/or treatment of diabetic complications. PMID:25255750

  19. Osmotic Stress, not Aldose Reductase Activity, Directly induces Growth Factors and MAPK Signaling changes during Sugar Cataract Formation

    PubMed Central

    Zhang, Peng; Xing, Kuiyi; Randazzo, James; Blessing, Karen; Lou, Marjorie F.; Kador, Peter F.

    2012-01-01

    In sugar cataract formation in rats, aldose reductase (AR) actitvity is not only linked to lenticular sorbitol (diabetic) or galactitol (galactosemic) formation but also to signal transduction changes, cytotoxic signals and activation of apoptosis. Using both in vitro and in vivo techniques, the interrelationship between AR activity, polyol (sorbitol and galactitol) formation, osmotic stress, growth factor induction, and cell signaling changes have been investigated. For in vitro studies, lenses from Sprague Dawley rats were cultured for up to 48 hrs in TC-199-bicarbonate media containing either 30 mM fructose (control), or 30 mM glucose or galctose with/without the aldose reductase inhibitors AL1576 or tolrestat, the sorbitol dehydrogenase inhibitor (SDI) CP-470,711, or 15 mM mannitol (osmotic-compensated media). For in vivo studies, lenses were obtained from streptozotocin-induced diabetic Sprague Dawley rats fed diet with/without the ARIs AL1576 or tolrestat for 10 weeks. As expected, lenses cultured in high glucose / galactose media or from untreated diabetic rats all showed a decrease in the GSH pool that was lessened by ARI treatment. Lenses either from diabetic rats or from glucose/galactose culture conditions showed increased expression of basic-FGF, TGF-β, and increased signaling through P-Akt, P-ERK1/2 and P-SAPK/JNK which were also normalized by ARIs to the expression levels observed in non-diabetic controls. Culturing rat lenses in osomotically compensated media containing 30 mM glucose or galactose did not lead to increased growth factor expression or altered signaling. These studies indicate that it is the biophysical response of the lens to osmotic stress that results in an increased intralenticular production of basic-FGF and TGF-β and the altered cytotoxic signaling that is observed during sugar cataract formation. PMID:22710095

  20. Esculetin, a Coumarin Derivative, Inhibits Aldose Reductase Activity in vitro and Cataractogenesis in Galactose-Fed Rats

    PubMed Central

    Kim, Chan-Sik; Kim, Junghyun; Lee, Yun Mi; Sohn, Eunjin; Kim, Jin Sook

    2016-01-01

    Naturally occurring coumarin compounds have received substantial attention due to their pharmaceutical effects. Esculetin is a coumarin derivative and a polyphenol compound that is used in a variety of therapeutic and pharmacological strategies. However, its effect on aldose reductase activity remains poorly understood. In this study, the potential beneficial effects of esculetin on lenticular aldose reductase were investigated in galactose-fed (GAL) rats, an animal model of sugar cataracts. Cataracts were induced in Sprague-Dawley (SD) rats via a 50% galactose diet for 2 weeks, and groups of GAL rats were orally treated with esculetin (10 or 50 mg/kg body weight). In vehicle-treated GAL rats, lens opacification was observed, and swelling and membrane rupture of the lens fiber cells were increased. Additionally, aldose reductase was highly expressed in the lens epithelium and superficial cortical fibers during cataract development in the GAL rats. Esculetin reduced rat lens aldose reductase (RLAR) activity in vitro, and esculetin treatment significantly inhibited lens opacity, as well as morphological alterations, such as swelling, vacuolation and liquefaction of lens fibers, via the inhibition of aldose reductase in the GAL rats. These results indicate that esculetin is a useful treatment for galactose-induced cataracts. PMID:26902086

  1. Esculetin, a Coumarin Derivative, Inhibits Aldose Reductase Activity in vitro and Cataractogenesis in Galactose-Fed Rats.

    PubMed

    Kim, Chan-Sik; Kim, Junghyun; Lee, Yun Mi; Sohn, Eunjin; Kim, Jin Sook

    2016-03-01

    Naturally occurring coumarin compounds have received substantial attention due to their pharmaceutical effects. Esculetin is a coumarin derivative and a polyphenol compound that is used in a variety of therapeutic and pharmacological strategies. However, its effect on aldose reductase activity remains poorly understood. In this study, the potential beneficialeffects of esculetin on lenticular aldose reductase were investigated in galactose-fed (GAL) rats, an animal model of sugar cataracts. Cataracts were induced in Sprague-Dawley (SD) rats via a 50% galactose diet for 2 weeks, and groups of GAL rats were orally treated with esculetin (10 or 50 mg/kg body weight). In vehicle-treated GAL rats, lens opacificationwas observed, and swelling and membrane rupture of the lens fibercells were increased. Additionally, aldose reductase was highly expressed in the lens epithelium and superficialcortical fibersduring cataract development in the GAL rats. Esculetin reduced rat lens aldose reductase (RLAR) activity in vitro, and esculetin treatment significanty inhibited lens opacity, as well as morphological alterations, such as swelling, vacuolation and liquefaction of lens fibers,via the inhibition of aldose reductase in the GAL rats. These results indicate that esculetin is a useful treatment for galactose-induced cataracts. PMID:26902086

  2. Inhibition of aldose reductase and anti-cataract action of trans-anethole isolated from Foeniculum vulgare Mill. fruits.

    PubMed

    Dongare, Vandana; Kulkarni, Chaitanya; Kondawar, Manish; Magdum, Chandrakant; Haldavnekar, Vivek; Arvindekar, Akalpita

    2012-05-01

    Foeniculum vulgare fruits are routinely consumed for their carminative and mouth freshening effect. The plant was evaluated for aldose reductase inhibition and anti-diabetic action. Bioguided fractionation using silica gel column chromatography, HPLC, and GC-MS analysis revealed trans-anethole as the bioactive constituent possessing potent aldose reductase inhibitory action, with an IC50 value of 3.8μg/ml. Prolonged treatment with the pet ether fraction of the F. vulgare distillate demonstrated improvement in blood glucose, lipid profile, glycated haemoglobin and other parameters in streptozotocin-induced diabetic rats. Trans-anethole could effectively show anti-cataract activity through the increase in soluble lens protein, reduced glutathione, catalase and SOD activity on in vitro incubation of the eye lens with 55mM glucose. Trans-anethole demonstrated noncompetitive to mixed type of inhibition of lens aldose reductase using Lineweaver Burk plot.

  3. Quantum Model of Catalysis Based on a Mobile Proton Revealed by Subatomic X-ray and Neutron Diffraction Studies of h-aldose Reductase

    SciTech Connect

    Blakeley, M. P.; Ruiz, Fredrico; Cachau, Raul; Hazemann, I.; Meilleur, Flora; Mitschler, A.; Ginell, Stephan; Afonine, Pavel; Ventura, Oscar; Cousido-Siah, Alexandra; Haertlein, M.; Joachimiak, Andrzej; Myles, Dean A A; Podjarny, A.

    2008-01-01

    We present results of combined studies of the enzyme human aldose reductase (h-AR, 36 kDa) using single-crystal x-ray data (0.66 Angstroms, 100K; 0.80 Angstroms, 15K; 1.75 Angstroms, 293K), neutron Laue data (2.2 Angstroms, 293K), and quantum mechanical modeling. These complementary techniques unveil the internal organization and mobility of the hydrogen bond network that defines the properties of the catalytic engine, explaining how this promiscuous enzyme overcomes the simultaneous requirements of efficiency and promiscuity offering a general mechanistic view for this class of enzymes.

  4. Amelioration of Acute Kidney Injury in Lipopolysaccharide-Induced Systemic Inflammatory Response Syndrome by an Aldose Reductase Inhibitor, Fidarestat

    PubMed Central

    Takahashi, Kazunori; Mizukami, Hiroki; Kamata, Kosuke; Inaba, Wataru; Kato, Noriaki; Hibi, Chihiro; Yagihashi, Soroku

    2012-01-01

    Background Systemic inflammatory response syndrome is a fatal disease because of multiple organ failure. Acute kidney injury is a serious complication of systemic inflammatory response syndrome and its genesis is still unclear posing a difficulty for an effective treatment. Aldose reductase (AR) inhibitor is recently found to suppress lipopolysaccharide (LPS)-induced cardiac failure and its lethality. We studied the effects of AR inhibitor on LPS-induced acute kidney injury and its mechanism. Methods Mice were injected with LPS and the effects of AR inhibitor (Fidarestat 32 mg/kg) before or after LPS injection were examined for the mortality, severity of renal failure and kidney pathology. Serum concentrations of cytokines (interleukin-1β, interleukin-6, monocyte chemotactic protein-1 and tumor necrosis factor-α) and their mRNA expressions in the lung, liver, spleen and kidney were measured. We also evaluated polyol metabolites in the kidney. Results Mortality rate within 72 hours was significantly less in LPS-injected mice treated with AR inhibitor both before (29%) and after LPS injection (40%) than untreated mice (90%). LPS-injected mice showed marked increases in blood urea nitrogen, creatinine and cytokines, and AR inhibitor treatment suppressed the changes. LPS-induced acute kidney injury was associated with vacuolar degeneration and apoptosis of renal tubular cells as well as infiltration of neutrophils and macrophages. With improvement of such pathological findings, AR inhibitor treatment suppressed the elevation of cytokine mRNA levels in multiple organs and renal sorbitol accumulation. Conclusion AR inhibitor treatment ameliorated LPS-induced acute kidney injury, resulting in the lowered mortality. PMID:22253906

  5. Aldose reductase (-106) C/T gene polymorphism and possibility of macrovascular complications in Egyptian type 2 diabetic patients

    PubMed Central

    Nomair, Azhar Mohamed; ElDeeb, Mona Mohamed Kamal; Maharem, Dalia Aly

    2016-01-01

    Introduction: Over the past three decades, the number of people with diabetes mellitus (DM) has more than doubled globally, making it one of the most important public health challenges to all nations. Aldose reductase (AR) is a rate-limiting enzyme in the polyol pathway, which has been implicated in the pathogenesis of diabetic microvascular complications; however, the association of the AR gene with diabetic macrovascular complications has rarely been investigated. Aim: The study aimed to identify the possible association between C(-106) T polymorphism of the AR gene and diabetic macroangiopathy in a cohort of Egyptian patients with type 2 DM. Settings and Design: This study was conducted on 100 Egyptian subjects, the control group (n = 20) and the patient group (n = 80) with type 2 diabetes which were further subdivided into two subgroups with (n = 48) and without macroangiopathic complications (n = 32) as evidenced by carotid intima-media thickness, electrocardiography (ECG) ischemic changes, cerebrovascular insufficiency, and peripheral vascular insufficiency. Subjects and Methods: All studied subjects were subjected to detailed history taking, clinical examination, ECG, carotid ultrasonography, routine laboratory investigations, and molecular studies including the detection of AR C(-106) T gene polymorphisms using the polymerase chain reaction (PCR)/restriction fragment length polymorphism technique. Results: The genotype distribution and allele frequency of AR C(-106) T showed no statistical significance also the genotypes were not associated with any of the different studied parameters. Conclusions: The results suggest that the C(-106) T polymorphism in the AR gene is not involved in the pathogenesis of macroangiopathy in type 2 diabetes. PMID:27730075

  6. GP-1447, an inhibitor of aldose reductase, prevents the progression of diabetic cataract in rats.

    PubMed

    Kawakubo, Ken; Mori, Asami; Sakamoto, Kenji; Nakahara, Tsutomu; Ishii, Kunio

    2012-01-01

    We examined the effects of GP-1447 (3-[(4,5,7-trifluorobenzothiazol-2-yl)methyl]-5-methylphenyl acetic acid) on existing cataracts and sorbitol content in the lens in rats with streptozotocin-induced diabetes. GP-1447 is an inhibitor of aldose reductase, which is the first enzyme in the polyol pathway. Cataracts in the central region of the lens were observed in 7 of 14 eyes (50%) by the fifth week after induction of diabetes, and development of mature cataracts was observed in most lenses by the ninth week. In diabetic rats that received GP-1447 treatment beginning in the fifth week after induction of diabetes, progression of cataracts was observed for 1 week after initiation of treatment. Thereafter, the severity of cataracts did not change substantially. Sorbitol levels in the lens peaked during the first week of diabetes, and this increase was maintained during the 9-week observation period. Elevated sorbitol levels in the lenses of diabetic rats gradually declined after GP-1447 treatment was started on the fifth week after induction of diabetes. Cataracts and sorbitol elevation were not observed in the lenses of controls or diabetic rats treated with GP-1447 immediately after induction of diabetes. These results suggest that the polyol pathway plays an important role in both the appearance and progression of cataracts in diabetic rats. Inhibition of aldose reductase could significantly prevent progression of existing cataracts. PMID:22687477

  7. Phytochemical analysis with the antioxidant and aldose reductase inhibitory capacities of Tephrosia humilis aerial parts' extracts.

    PubMed

    Plioukas, Michael; Gabrieli, Chrysi; Lazari, Diamanto; Kokkalou, Eugene

    2016-06-01

    The aerial parts of Tephrosia humilis were tested about their antioxidant potential, their ability to inhibit the aldose/aldehyde reductase enzymes and their phenolic content. The plant material was exhaustively extracted with petroleum ether, dichloromethane and methanol, consecutively. The concentrated methanol extract was re-extracted, successively, with diethyl ether, ethyl acetate and n-butanol. All extracts showed significant antioxidant capacity, but the most effective was the ethyl acetate extract. As about the aldose reductase inhibition, all fractions, except the aqueous, were strong inhibitors of the enzyme, with the n-butanolic and ethyl acetate fractions to inhibit the enzyme above 75%. These findings provide support to the ethnopharmacological usage of the plant as antioxidant and validate its potential to act against the long-term diabetic complications. The phytochemical analysis showed the presence of 1,4-dihydroxy-3,4-(epoxyethano)-5-cyclohexene(1), cleroindicin E(2), lupeol(3), methyl p-coumarate(4), methyl 4-hydroxybenzoate(5), prunin(6), 5,7,2',5'-tetrahydroxyflavanone 7-rutinoside(7), protocatechuic acid(8), luteolin 7-glucoside(9), apigenin(10), naringin(11), rhoifolin(12) and luteolin 7-glucuronate(13).

  8. Phytochemical analysis with the antioxidant and aldose reductase inhibitory capacities of Tephrosia humilis aerial parts' extracts.

    PubMed

    Plioukas, Michael; Gabrieli, Chrysi; Lazari, Diamanto; Kokkalou, Eugene

    2016-06-01

    The aerial parts of Tephrosia humilis were tested about their antioxidant potential, their ability to inhibit the aldose/aldehyde reductase enzymes and their phenolic content. The plant material was exhaustively extracted with petroleum ether, dichloromethane and methanol, consecutively. The concentrated methanol extract was re-extracted, successively, with diethyl ether, ethyl acetate and n-butanol. All extracts showed significant antioxidant capacity, but the most effective was the ethyl acetate extract. As about the aldose reductase inhibition, all fractions, except the aqueous, were strong inhibitors of the enzyme, with the n-butanolic and ethyl acetate fractions to inhibit the enzyme above 75%. These findings provide support to the ethnopharmacological usage of the plant as antioxidant and validate its potential to act against the long-term diabetic complications. The phytochemical analysis showed the presence of 1,4-dihydroxy-3,4-(epoxyethano)-5-cyclohexene(1), cleroindicin E(2), lupeol(3), methyl p-coumarate(4), methyl 4-hydroxybenzoate(5), prunin(6), 5,7,2',5'-tetrahydroxyflavanone 7-rutinoside(7), protocatechuic acid(8), luteolin 7-glucoside(9), apigenin(10), naringin(11), rhoifolin(12) and luteolin 7-glucuronate(13). PMID:26209262

  9. Inhibitory effects of Zingiber officinale Roscoe derived components on aldose reductase activity in vitro and in vivo.

    PubMed

    Kato, Atsushi; Higuchi, Yasuko; Goto, Hirozo; Kizu, Haruhisa; Okamoto, Tadashi; Asano, Naoki; Hollinshead, Jackie; Nash, Robert J; Adachi, Isao

    2006-09-01

    Ginger (Zingiber officinale Roscoe) continues to be used as an important cooking spice and herbal medicine around the world. Scientific research has gradually verified the antidiabetic effects of ginger. Especially gingerols, which are the major components of ginger, are known to improve diabetes including the effect of enhancement against insulin-sensitivity. Aldose reductase inhibitors have considerable potential for the treatment of diabetes, without increased risk of hypoglycemia. The assay for aldose reductase inhibitors in ginger led to the isolation of five active compounds including 2-(4-hydroxy-3-methoxyphenyl)ethanol (2) and 2-(4-hydroxy-3-methoxyphenyl)ethanoic acid (3). Compounds 2 and 3 were good inhibitors of recombinant human aldose reductase, with IC50 values of 19.2 +/- 1.9 and 18.5 +/- 1.1 microM, respectively. Furthermore, these compounds significantly suppressed not only sorbitol accumulation in human erythrocytes but also lens galactitol accumulation in 30% of galactose-fed cataract rat model. A structure-activity relationship study revealed that the applicable side alkyl chain length and the presence of a C3 OCH3 group in the aromatic ring are essential features for enzyme recognition and binding. These results suggested that it would contribute to the protection against or improvement of diabetic complications for a dietary supplement of ginger or its extract containing aldose reductase inhibitors. PMID:16939321

  10. Model of the catalytic mechanism of human aldose reductase based on quantum chemical calculations.

    SciTech Connect

    Cachau, R. C.; Howard, E. H.; Barth, P. B.; Mitschler, A. M.; Chevrier, B. C.; Lamour, V.; Joachimiak, A.; Sanishvili, R.; Van Zandt, M.; Sibley, E.; Moras, D.; Podjarny, A.; UPR de Biologie Structurale; National Cancer Inst.; Univ. Louis Pasteur; Inst. for Diabetes Discovery, Inc.

    2000-01-01

    Aldose Reductase is an enzyme involved in diabetic complications, thoroughly studied for the purpose of inhibitor development. The structure of an enzyme-inhibitor complex solved at sub-atomic resolution has been used to develop a model for the catalytic mechanism. This model has been refined using a combination of Molecular Dynamics and Quantum calculations. It shows that the proton donation, the subject of previous controversies, is the combined effect of three residues: Lys 77, Tyr 48 and His 110. Lys 77 polarises the Tyr 48 OH group, which donates the proton to His 110, which becomes doubly protonated. His 110 then moves and donates the proton to the substrate. The key information from the sub-atomic resolution structure is the orientation of the ring and the single protonafion of the His 110 in the enzyme-inhibitor complex. This model is in full agreement with all available experimental data.

  11. SIRT6 Is a Positive Regulator of Aldose Reductase Expression in U937 and HeLa cells under Osmotic Stress: In Vitro and In Silico Insights

    PubMed Central

    Timucin, Ahmet Can; Basaga, Huveyda

    2016-01-01

    SIRT6 is a protein deacetylase, involved in various intracellular processes including suppression of glycolysis and DNA repair. Aldose Reductase (AR), first enzyme of polyol pathway, was proposed to be indirectly associated to these SIRT6 linked processes. Despite these associations, presence of SIRT6 based regulation of AR still remains ambiguous. Thus, regulation of AR expression by SIRT6 was investigated under hyperosmotic stress. A unique model of osmotic stress in U937 cells was used to demonstrate the presence of a potential link between SIRT6 and AR expression. By overexpressing SIRT6 in HeLa cells under hyperosmotic stress, its role on upregulation of AR was revealed. In parallel, increased SIRT6 activity was shown to upregulate AR in U937 cells under hyperosmotic milieu by using pharmacological modulators. Since these modulators also target SIRT1, binding of the inhibitor, Ex-527, specifically to SIRT6 was analyzed in silico. Computational observations indicated that Ex-527 may also target SIRT6 active site residues under high salt concentration, thus, validating in vitro findings. Based on these evidences, a novel regulatory step by SIRT6, modifying AR expression under hyperosmotic stress was presented and its possible interactions with intracellular machinery was discussed. PMID:27536992

  12. X-ray structure of the V301L aldo-keto reductase 1B10 complexed with NADP(+) and the potent aldose reductase inhibitor fidarestat: implications for inhibitor binding and selectivity.

    PubMed

    Ruiz, Francesc Xavier; Cousido-Siah, Alexandra; Mitschler, André; Farrés, Jaume; Parés, Xavier; Podjarny, Alberto

    2013-02-25

    Only one crystal structure is currently available for tumor marker AKR1B10, complexed with NADP(+) and tolrestat, which is an aldose reductase inhibitor (ARI) of the carboxylic acid type. Here, the X-ray structure of the complex of the V301L substituted AKR1B10 holoenzyme with fidarestat, an ARI of the cyclic imide type, was obtained at 1.60Å resolution by replacement soaking of crystals containing tolrestat. Previously, fidarestat was found to be safe in phase III trials for diabetic neuropathy and, consistent with its low in vivo side effects, was highly selective for aldose reductase (AR or AKR1B1) versus aldehyde reductase (AKR1A1). Now, inhibition studies showed that fidarestat was indeed 1300-fold more selective for AR as compared to AKR1B10, while the change of Val to Leu (found in AR) caused a 20-fold decrease in the IC50 value with fidarestat. Structural analysis of the V301L AKR1B10-fidarestat complex displayed enzyme-inhibitor interactions similar to those of the AR-fidarestat complex. However, a close inspection of both the new crystal structure and a computer model of the wild-type AKR1B10 complex with fidarestat revealed subtle changes that could affect fidarestat binding. In the crystal structure, a significant motion of loop A was observed between AR and V301L AKR1B10, linked to a Phe-122/Phe-123 side chain displacement. This was due to the presence of the more voluminous Gln-303 side chain (Ser-302 in AR) and of a water molecule buried in a subpocket located at the base of flexible loop A. In the wild-type AKR1B10 model, a short contact was predicted between the Val-301 side chain and fidarestat, but would not be present in AR or in V301L AKR1B10. Overall, these changes could contribute to the difference in inhibitory potency of fidarestat between AR and AKR1B10.

  13. Construction of an Indonesian herbal constituents database and its use in Random Forest modelling in a search for inhibitors of aldose reductase.

    PubMed

    Naeem, Sadaf; Hylands, Peter; Barlow, David

    2012-02-01

    Data on phytochemical constituents of plants commonly used in traditional Indonesian medicine have been compiled as a computer database. This database (the Indonesian Herbal constituents database, IHD) currently contains details on ∼1,000 compounds found in 33 different plants. For each entry, the IHD gives details of chemical structure, trivial and systematic name, CAS registry number, pharmacology (where known), toxicology (LD(50)), botanical species, the part(s) of the plant(s) where the compounds are found, typical dosage(s) and reference(s). A second database has been also been compiled for plant-derived compounds with known activity against the enzyme, aldose reductase (AR). This database (the aldose reductase inhibitors database, ARID) contains the same details as the IHD, and currently comprises information on 120 different AR inhibitors. Virtual screening of all compounds in the IHD has been performed using Random Forest (RF) modelling, in a search for novel leads active against AR-to provide for new forms of symptomatic relief in diabetic patients. For the RF modelling, a set of simple 2D chemical descriptors were employed to classify all compounds in the combined ARID and IHD databases as either active or inactive as AR inhibitors. The resulting RF models (which gave misclassification rates of 21%) were used to identify putative new AR inhibitors in the IHD, with such compounds being identified as those giving RF scores >0.5 (in each of the three different RF models developed). In vitro assays were subsequently performed for four of the compounds obtained as hits in this in silico screening, to determine their inhibitory activity against human recombinant AR. The two compounds having the highest RF scores (prunetin and ononin) were shown to have the highest activities experimentally (giving ∼58% and ∼52% inhibition at a concentration of 15μM, respectively), while the compounds with lowest RF scores (vanillic acid and cinnamic acid) showed the

  14. Structure-activity relationships and molecular modelling of new 5-arylidene-4-thiazolidinone derivatives as aldose reductase inhibitors and potential anti-inflammatory agents.

    PubMed

    Maccari, Rosanna; Vitale, Rosa Maria; Ottanà, Rosaria; Rocchiccioli, Marco; Marrazzo, Agostino; Cardile, Venera; Graziano, Adriana Carol Eleonora; Amodeo, Pietro; Mura, Umberto; Del Corso, Antonella

    2014-06-23

    A series of 5-(carbamoylmethoxy)benzylidene-2-oxo/thioxo-4-thiazolidinone derivatives (6-9) were synthesized as inhibitors of aldose reductase (AR), enzyme which plays a crucial role in the development of diabetes complications as well as in the inflammatory processes associated both to diabetes mellitus and to other pathologies. In vitro inhibitory activity indicated that compounds 6-9a-d were generally good AR inhibitors. Acetic acid derivatives 8a-d and 9a-d were shown to be the best enzyme inhibitors among the tested compounds endowed with significant inhibitory ability levels reaching submicromolar IC50 values. Moreover, some representative AR inhibitors (7a, 7c, 9a, 9c, 9d) were assayed in cultures of human keratinocytes in order to evaluate their capability to reduce NF-kB activation and iNOS expression. Compound 9c proved to be the best derivative endowed with both interesting AR inhibitory effectiveness and ability to reduce NF-kB activation and iNOS expression. Molecular docking and molecular dynamics simulations were undertaken to investigate the binding modes of selected compounds into the active site of AR in order to rationalize the inhibitory effectiveness of these derivatives.

  15. Stress tolerance of transgenic barley accumulating the alfalfa aldose reductase in the cytoplasm and the chloroplast.

    PubMed

    Nagy, Bettina; Majer, Petra; Mihály, Róbert; Pauk, János; Horváth, Gábor V

    2016-09-01

    Barley represents one of the major crops grown worldwide; its genetic transformation provides an important tool for the improvement of crop quality and tolerance to environmental stress factors. Biotic and abiotic stresses produce reactive oxygen species in the plant cells that can directly oxidize the cellular components including lipid membranes; resulting in lipid peroxidation and subsequently the accumulation of reactive carbonyl compounds. In order to protect barley plants from the effects of stress-produced reactive carbonyls, an Agrobacterium-mediated transformation was carried out using the Medicago sativa aldose reductase (MsALR) gene. In certain transgenic lines the produced MsALR enzyme was targeted to the chloroplasts to evaluate its protective effect in these organelles. The dual fluorescent protein-based method was used for the evaluation of tolerance of young seedlings to diverse stresses; our results demonstrated that this technique could be reliably applied for the detection of cellular stress in a variety of conditions. The chlorophyll and carotenoid content measurements also supported the results of the fluorescent protein-based method and the stress-protective effect of the MsALR enzyme. Targeting of MsALR into the chloroplast has also resulted in increased stress tolerance, similarly to the observed effect of the cytosolic MsALR accumulation. The results of the DsRed/GFP fluorescent protein-based method indicated that both the cytosol and chloroplast accumulation of MsALR can increase the abiotic stress tolerance of transgenic barley lines. PMID:27469099

  16. Design and synthesis of potent and multifunctional aldose reductase inhibitors based on quinoxalinones.

    PubMed

    Qin, Xiangyu; Hao, Xin; Han, Hui; Zhu, Shaojuan; Yang, Yanchun; Wu, Bobin; Hussain, Saghir; Parveen, Shagufta; Jing, Chaojun; Ma, Bing; Zhu, Changjin

    2015-02-12

    Quinoxalin-2(1H)-one based design and synthesis produced several series of aldose reductase (ALR2) inhibitor candidates. In particular, phenolic structure was installed in the compounds for the combination of antioxidant activity and strengthening the ability to fight against diabetic complications. Most of the series 6 showed potent and selective effects on ALR2 inhibition with IC50 values in the range of 0.032-0.468 μM, and 2-(3-(2,4-dihydroxyphenyl)-7-fluoro-2-oxoquinoxalin-1(2H)-yl)acetic acid (6e) was the most active. More significantly, most of the series 8 revealed not only good activity in the ALR2 inhibition but also potent antioxidant activity, and 2-(3-(3-methoxy-4-hydroxystyryl)-2-oxoquinoxalin-1(2H)-yl)acetic acid (8d) was even as strong as the well-known antioxidant Trolox at a concentration of 100 μM, verifying the C3 p-hydroxystyryl side chain as the key structure for alleviating oxidative stress. These results therefore suggest an achievement of multifunctional ALR2 inhibitors having both potency for ALR2 inhibition and as antioxidants. PMID:25602762

  17. Glucose and collagen regulate human platelet activity through aldose reductase induction of thromboxane.

    PubMed

    Tang, Wai Ho; Stitham, Jeremiah; Gleim, Scott; Di Febbo, Concetta; Porreca, Ettore; Fava, Cristiano; Tacconelli, Stefania; Capone, Marta; Evangelista, Virgilio; Levantesi, Giacomo; Wen, Li; Martin, Kathleen; Minuz, Pietro; Rade, Jeffrey; Patrignani, Paola; Hwa, John

    2011-11-01

    Diabetes mellitus is associated with platelet hyperactivity, which leads to increased morbidity and mortality from cardiovascular disease. This is coupled with enhanced levels of thromboxane (TX), an eicosanoid that facilitates platelet aggregation. Although intensely studied, the mechanism underlying the relationship among hyperglycemia, TX generation, and platelet hyperactivity remains unclear. We sought to identify key signaling components that connect high levels of glucose to TX generation and to examine their clinical relevance. In human platelets, aldose reductase synergistically modulated platelet response to both hyperglycemia and collagen exposure through a pathway involving ROS/PLCγ2/PKC/p38α MAPK. In clinical patients with platelet activation (deep vein thrombosis; saphenous vein graft occlusion after coronary bypass surgery), and particularly those with diabetes, urinary levels of a major enzymatic metabolite of TX (11-dehydro-TXB2 [TX-M]) were substantially increased. Elevated TX-M persisted in diabetic patients taking low-dose aspirin (acetylsalicylic acid, ASA), suggesting that such patients may have underlying endothelial damage, collagen exposure, and thrombovascular disease. Thus, our study has identified multiple potential signaling targets for designing combination chemotherapies that could inhibit the synergistic activation of platelets by hyperglycemia and collagen exposure. PMID:22005299

  18. Aldose Reductase Regulates Microglia/Macrophages Polarization Through the cAMP Response Element-Binding Protein After Spinal Cord Injury in Mice.

    PubMed

    Zhang, Qian; Bian, Ganlan; Chen, Peng; Liu, Ling; Yu, Caiyong; Liu, Fangfang; Xue, Qian; Chung, Sookja K; Song, Bing; Ju, Gong; Wang, Jian

    2016-01-01

    Inflammatory reactions are the most critical pathological processes occurring after spinal cord injury (SCI). Activated microglia/macrophages have either detrimental or beneficial effects on neural regeneration based on their functional polarized M1/M2 subsets. However, the mechanism of microglia/macrophage polarization to M1/M2 at the injured spinal cord environment remains unknown. In this study, wild-type (WT) or aldose reductase (AR)-knockout (KO) mice were subjected to SCI by a spinal crush injury model. The expression pattern of AR, behavior tests for locomotor activity, and lesion size were assessed at between 4 h and 28 days after SCI. We found that the expression of AR is upregulated in microglia/macrophages after SCI in WT mice. In AR KO mice, SCI led to smaller injury lesion areas compared to WT. AR deficiency-induced microglia/macrophages induce the M2 rather than the M1 response and promote locomotion recovery after SCI in mice. In the in vitro experiments, microglia cell lines (N9 or BV2) were treated with the AR inhibitor (ARI) fidarestat. AR inhibition caused 4-hydroxynonenal (HNE) accumulation, which induced the phosphorylation of the cAMP response element-binding protein (CREB) to promote Arg1 expression. KG501, the specific inhibitor of phosphorylated CREB, could cancel the upregulation of Arg1 by ARI or HNE stimulation. Our results suggest that AR works as a switch which can regulate microglia by polarizing cells to either the M1 or the M2 phenotype under M1 stimulation based on its states of activity. We suggest that inhibiting AR may be a promising therapeutic method for SCI in the future.

  19. Prospecting for Novel Plant-Derived Molecules of Rauvolfia serpentina as Inhibitors of Aldose Reductase, a Potent Drug Target for Diabetes and Its Complications

    PubMed Central

    Pathania, Shivalika; Randhawa, Vinay; Bagler, Ganesh

    2013-01-01

    Aldose Reductase (AR) is implicated in the development of secondary complications of diabetes, providing an interesting target for therapeutic intervention. Extracts of Rauvolfia serpentina, a medicinal plant endemic to the Himalayan mountain range, have been known to be effective in alleviating diabetes and its complications. In this study, we aim to prospect for novel plant-derived inhibitors from R. serpentina and to understand structural basis of their interactions. An extensive library of R. serpentina molecules was compiled and computationally screened for inhibitory action against AR. The stability of complexes, with docked leads, was verified using molecular dynamics simulations. Two structurally distinct plant-derived leads were identified as inhibitors: indobine and indobinine. Further, using these two leads as templates, 16 more leads were identified through ligand-based screening of their structural analogs, from a small molecules database. Thus, we obtained plant-derived indole alkaloids, and their structural analogs, as potential AR inhibitors from a manually curated dataset of R. serpentina molecules. Indole alkaloids reported herein, as a novel structural class unreported hitherto, may provide better insights for designing potential AR inhibitors with improved efficacy and fewer side effects. PMID:23613832

  20. Probenecid Treatment Enhances Retinal and Brain Delivery of N-4-Benzoylaminophenylsulfonylglycine, An Anionic Aldose Reductase Inhibitor

    PubMed Central

    Sunkara, Gangadhar; Ayalasomayajula, Surya P.; DeRuiter, Jack; Kompella, Uday B.

    2009-01-01

    Anion efflux transporters are expected to minimize target tissue delivery of N-[4-(benzoylaminophenyl)sulfonyl]glycine (BAPSG), a novel carboxylic acid aldose reductase inhibitor, which exists as a monocarboxylate anion at physiological conditions. Therefore, the objective of this study was to determine whether BAPSG delivery to various eye tissues including the retina and the brain can be enhanced by probenecid, a competitive inhibitor of anion transporters. To determine the influence of probenecid on eye and brain distribution of BAPSG, probenecid was administered intraperitoneally (120 mg/kg body weight; i.p.) 20 minutes prior to BAPSG (50 mg/kg; i.p.) administration. Drug disposition in various eye tissues including the retina and the brain was determined at 15 min, 1, 2 and 4 hr after BAPSG dose in male Sprauge-Dawley rats. To determine whether probenecid alters plasma clearance of BAPSG, influence of probenecid (120 mg/kg; i.p.) on the plasma pharmacokinetics of intravenously administered BAPSG (15 mg/kg) was studied as well. Finally, the effect of probenecid co-administration on the ocular tissue distribution of BAPSG was assessed in rabbits following topical (eye drop) administration. Following pretreatment with probenecid in the rat study, retinal delivery at 1 hr was increased by about 11 fold (2580 vs 244 ng/gm; p<0.05). Further, following probenecid pretreatment, significant BAPSG levels were detectable in the brain (45 ± 20 ng/gm) at 1 hr, unlike controls where the drug was not detectable. Plasma concentrations, plasma elimination half-life, and total body clearance of intravenously administered BAPSG were not altered by i.p. probenecid pretreatment. In the topical dosing study, a significant decline in BAPSG delivery was observed in the iris-ciliary body but no significant changes were observed in other tissues of the anterior segment of the eye including tears. Thus, inhibition of anion transporters is a useful approach to elevate retinal and brain

  1. Tonicity-responsive enhancer binding protein regulates the expression of aldose reductase and protein kinase C δ in a mouse model of diabetic retinopathy.

    PubMed

    Park, Jeongsook; Kim, Hwajin; Park, So Yun; Lim, Sun Woo; Kim, Yoon Sook; Lee, Dong Hoon; Roh, Gu Seob; Kim, Hyun Joon; Kang, Sang Soo; Cho, Gyeong Jae; Jeong, Bo-Young; Kwon, H Moo; Choi, Wan Sung

    2014-05-01

    Recent studies revealed that Tonicity-responsive enhancer binding protein (TonEBP) directly regulates the transcription of aldose reductase (AR), which catalyzes the first step of the polyol pathway of glucose metabolism. Activation of protein kinase C δ (PKCδ) is dependent on AR and it has been linked to diabetic complications. However, whether TonEBP affects expressions of AR and PKCδ in diabetic retinopathy was not clearly shown. In this study, we used TonEBP heterozygote mice to study the role of TonEBP in streptozotocin (STZ)-induced diabetic retinopathy. We performed immunofluorescence staining and found that retinal expressions of AR and PKCδ were significantly reduced in the heterozygotes compared to wild type littermates, particularly in ganglion cell layer. To examine further the effect of TonEBP reduction in retinal tissues, we performed intravitreal injection of TonEBP siRNA and confirmed the decrease in AR and PKCδ levels. In addition, we found that a proapoptotic factor, Bax level was reduced and a survival factor, Bcl2 level was increased after injection of TonEBP siRNA, indicating that TonEBP mediates apoptotic cell death. In parallel, TonEBP siRNA was applied to the in vitro human retinal pigment epithelial (ARPE-19) cells cultured in high glucose media. We have consistently found the decrease in AR and PKCδ levels and changes in apoptotic factors for survival. Together, these results clearly demonstrated that hyperglycemia-induced TonEBP plays a crucial role in increasing AR and PKCδ levels and leading to apoptotic death. Our findings suggest that TonEBP reduction is an effective therapeutic strategy for diabetic retinopathy. PMID:24631337

  2. Kinetic and molecular docking studies of loganin and 7-O-galloyl-D-sedoheptulose from Corni Fructus as therapeutic agents for diabetic complications through inhibition of aldose reductase.

    PubMed

    Lee, Chan Mee; Jung, Hyun Ah; Oh, Sang Ho; Park, Chan Hum; Tanaka, Takashi; Yokozawa, Takako; Choi, Jae Sue

    2015-06-01

    Aldose reductase (AR) is a key enzyme in the polyol pathway that is strongly implicated in the pathogenesis of diabetic complications. AR inhibitors have been proposed as therapeutic agents for diabetic complications through suppression of sorbitol formation and accumulation. In this study, we evaluated whether two major compounds of Corni Fructus, loganin and 7-O-galloyl-D-sedoheptulose, had an inhibitory effect on diabetic complications through AR inhibition. Because the iridoid glycoside loganin and the low-molecular-weight polyphenol 7-O-galloyl-D-sedoheptulose showed marginal inhibitory activities against rat lens AR (RLAR) and human recombinant AR (HRAR) in inhibition assays, we performed enzyme kinetic analyses and molecular simulation of the interaction of these two compounds with AR to further investigate their potential as inhibitors of diabetic complications. In kinetic analysis using Lineweaver-Burk plots and Dixon plots, loganin and 7-O-galloyl-D-sedoheptulose were both mixed inhibitors of RLAR with inhibition constants (K i) of 27.99 and 128.68 μΜ, respectively. Moreover, molecular docking simulation of both compounds demonstrated negative binding energies (Autodock 4.0 = -6.7; -7.5 kcal/mol; Fred 2.0 = -59.4; -63.2 kcal/mol) indicating a high affinity and tight binding capacity for the active site of the enzyme. Iridoid nucleus and aromatic ring systems and glycoside and sedoheptulose moieties were found to bind tightly to the specificity pocket and the anion binding pocket in RLAR through Phe123, His111, Trp21, Tyr49, His111, and Trp112 residues. Our results clearly indicate that loganin and 7-O-galloyl-D-sedoheptulose have great promise for the treatment of diabetic complications through inhibition of AR. PMID:25315636

  3. Kinetic and molecular docking studies of loganin and 7-O-galloyl-D-sedoheptulose from Corni Fructus as therapeutic agents for diabetic complications through inhibition of aldose reductase.

    PubMed

    Lee, Chan Mee; Jung, Hyun Ah; Oh, Sang Ho; Park, Chan Hum; Tanaka, Takashi; Yokozawa, Takako; Choi, Jae Sue

    2015-06-01

    Aldose reductase (AR) is a key enzyme in the polyol pathway that is strongly implicated in the pathogenesis of diabetic complications. AR inhibitors have been proposed as therapeutic agents for diabetic complications through suppression of sorbitol formation and accumulation. In this study, we evaluated whether two major compounds of Corni Fructus, loganin and 7-O-galloyl-D-sedoheptulose, had an inhibitory effect on diabetic complications through AR inhibition. Because the iridoid glycoside loganin and the low-molecular-weight polyphenol 7-O-galloyl-D-sedoheptulose showed marginal inhibitory activities against rat lens AR (RLAR) and human recombinant AR (HRAR) in inhibition assays, we performed enzyme kinetic analyses and molecular simulation of the interaction of these two compounds with AR to further investigate their potential as inhibitors of diabetic complications. In kinetic analysis using Lineweaver-Burk plots and Dixon plots, loganin and 7-O-galloyl-D-sedoheptulose were both mixed inhibitors of RLAR with inhibition constants (K i) of 27.99 and 128.68 μΜ, respectively. Moreover, molecular docking simulation of both compounds demonstrated negative binding energies (Autodock 4.0 = -6.7; -7.5 kcal/mol; Fred 2.0 = -59.4; -63.2 kcal/mol) indicating a high affinity and tight binding capacity for the active site of the enzyme. Iridoid nucleus and aromatic ring systems and glycoside and sedoheptulose moieties were found to bind tightly to the specificity pocket and the anion binding pocket in RLAR through Phe123, His111, Trp21, Tyr49, His111, and Trp112 residues. Our results clearly indicate that loganin and 7-O-galloyl-D-sedoheptulose have great promise for the treatment of diabetic complications through inhibition of AR.

  4. Chemical constituents from the aerial parts of Aster koraiensis with protein glycation and aldose reductase inhibitory activities.

    PubMed

    Lee, Jun; Lee, Yun Mi; Lee, Byong Won; Kim, Joo-Hwan; Kim, Jin Sook

    2012-02-24

    Two new eudesmane-type sesquiterpene glucosides, 9β-O-(E-p-hydroxycinnamoyl)-1β,6β-dihydroxy-trans-eudesm-3-en-6-O-β-D-glucopyranoside (1) and 9α-O-(E-p-hydroxycinnamoyl)-1α,6α-11-trihydroxy-trans-eudesm-3-en-6-O-β-D-glucopyranoside (2), were isolated by the activity-guidedfractionation of an EtOAc-soluble fraction from the aerial parts of Aster koraiensis. A new dihydrobenzofuran glucoside, (2R,3S)-6-acetyl-2-[1-O-(β-D-glucopyranosyl)-2-propenyl]-5-hydroxy-3-methoxy-2,3-dihydrobenzofuran (3), was also isolated, in addition to 15 known compounds. The structures of 1-3 were determined by spectroscopic data interpretation. All of the isolates were evaluated for in vitro inhibitory activity against the formation of advanced glycation end-products and rat lens aldose reductase.

  5. Coumarin-thiazole and -oxadiazole derivatives: Synthesis, bioactivity and docking studies for aldose/aldehyde reductase inhibitors.

    PubMed

    Ibrar, Aliya; Tehseen, Yildiz; Khan, Imtiaz; Hameed, Abdul; Saeed, Aamer; Furtmann, Norbert; Bajorath, Jürgen; Iqbal, Jamshed

    2016-10-01

    In continuation of our previous efforts directed towards the development of potent and selective inhibitors of aldose reductase (ALR2), and to control the diabetes mellitus (DM), a chronic metabolic disease, we synthesized novel coumarin-thiazole 6(a-o) and coumarin-oxadiazole 11(a-h) hybrids and screened for their inhibitory activity against aldose reductase (ALR2), for the selectivity against aldehyde reductase (ALR1). Compounds were also screened against ALR1. Among the newly designed compounds, 6c, 11d, and 11g were selective inhibitors of ALR2. Whereas, (E)-3-(2-(2-(2-bromobenzylidene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one 6c yielded the lowest IC50 value of 0.16±0.06μM for ALR2. Moreover, compounds (E)-3-(2-(2-benzylidenehydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6a; IC50=2.94±1.23μM for ARL1 and 0.12±0.05μM for ARL2) and (E)-3-(2-(2-(1-(4-bromophenyl)ethylidene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6e; IC50=1.71±0.01μM for ARL1 and 0.11±0.001μM for ARL2) were confirmed as dual inhibitors. Furthermore, compounds 6i, 6k, 6m, and 11b were found to be selective inhibitors for ALR1, among which (E)-3-(2-(2-((2-amino-4-chlorophenyl)(phenyl)methylene)hydrazinyl)thiazol-4-yl)-2H-chromen-2-one (6m) was most potent (IC50=0.459±0.001μM). Docking studies performed using X-ray structures of ALR1 and ALR2 with the given synthesized inhibitors showed that coumarinyl thiazole series lacks the carboxylate function that could interact with the anionic binding site being a common ALR1/ALR2 inhibitors trait. Molecular docking study with dual inhibitor 6e also suggested plausible binding modes for the ALR1 and ALR2 enzymes. Hence, the results of this study revealed that coumarinyl thiazole and oxadiazole derivatives could act as potential ALR1/ALR2 inhibitors.

  6. Bioactivity Focus of α-Cyano-4-hydroxycinnamic acid (CHCA) Leads to Effective Multifunctional Aldose Reductase Inhibitors

    PubMed Central

    Zhang, Laitao; Li, Yi-Fang; Yuan, Sheng; Zhang, Shijie; Zheng, Huanhuan; Liu, Jie; Sun, Pinghua; Gu, Yijun; Kurihara, Hiroshi; He, Rong-Rong; Chen, Heru

    2016-01-01

    Bioactivity focus on α-cyano-4-hydroxycinnamic acid (CHCA) scaffold results in a small library of novel multifunctional aldose reductase (ALR2) inhibitors. All the entities displayed good to excellent inhibition with IC50 72–405 nM. (R,E)-N-(3-(2-acetamido-3-(benzyloxy)propanamido)propyl)-2-cyano-3-(4-hydroxy phenyl)acrylamide (5f) was confirmed as the most active inhibitor (IC50 72.7 ± 1.6 nM), and the best antioxidant. 5f bound to ALR2 with new mode without affecting the aldehyde reductase (ALR1) activity, implicating high selectivity to ALR2. 5f was demonstrated as both an effective ALR2 inhibitor (ARI) and antioxidant in a chick embryo model of hyperglycemia. It attenuated hyperglycemia-induced incidence of neural tube defects (NTD) and death rate, and significantly improved the body weight and morphology of the embryos. 5f restored the expression of paired box type 3 transcription factor (Pax3), and reduced the hyperglycemia-induced increase of ALR2 activity, sorbitol accumulation, and the generation of ROS and MDA to normal levels. All the evidences support that 5f may be a potential agent to treat diabetic complications. PMID:27109517

  7. A novel zwitterionic inhibitor of aldose reductase interferes with polyol pathway in ex vivo and in vivo models of diabetic complications.

    PubMed

    Karasova, M Juskova; Prnova, M Soltesova; Stefek, M

    2014-10-01

    Recently a zwitterionic principle has been suggested as an alternative to bioisosteric replacement for increasing low bioavailability of aldose reductase inhibitors bearing an acidic function. In the present work we studied the effect of a novel zwitterionic inhibitor of aldose reductase [(2-benzyl-2,3,4,5-tetrahydro-1 H-pyrido[4,3-b]indole-8-yl)-acetic acid, compound 1] on sorbitol accumulation in ex vivo and in vivo models of diabetic complications. The effect of 1 on sorbitol accumulation in isolated rat eye lenses incubated with high glucose and in selected organs of streptozotocin-induced diabetic rats was evaluated. Significantly increased sorbitol levels were recorded in the lenses incubated with 50 mM glucose in comparison with controls. Sorbitol production was inhibited by 1 at concentrations of 25 and 100 μM. Under in vivo conditions in diabetic rats, significant elevation of sorbitol levels in selected organs was recorded. Compound 1 administered i.g. for five consecutive days (twice a day 25 mg/kg) inhibited sorbitol accumulation in erythrocytes and the sciatic nerve, yet it was without effect in eye lenses. A similar picture of inhibition was observed after i.p. administration of 1. To conclude, the results suggest that the zwitterionic principle may represent a practicable way of improving bioavailability of aldose reductase inhibitors bearing an acidic function.

  8. Galloyl glucoses from the seeds of Cornus officinalis with inhibitory activity against protein glycation, aldose reductase, and cataractogenesis ex vivo.

    PubMed

    Lee, Jun; Jang, Dae Sik; Kim, Nan Hee; Lee, Yun Mi; Kim, Junghyun; Kim, Jin Sook

    2011-01-01

    In an ongoing project directed toward the discovery of novel treatments for diabetic complications from traditional herbal medicines, six galloyl glucoses, 1,2,3-tri-O-galloyl-β-D-glucose (1), 1,2,6-tri-O-galloyl-β-D-glucose (2), 1,2,3,6-tetra-O-galloyl-β-D-glucose (3), 1,2,4,6-tetra-O-galloyl-β-D-glucose (4), 1,2,3,4,6-penta-O-galloyl-β-D-glucose (5), and tellimagrandin II (6), and two phenolic acids, gallic acid 4-O-β-D-glucoside (7) and gallic acid 4-O-β-D-(6'-O-galloyl)-glucoside (8), were isolated from an EtOAc-soluble fraction of the seeds of Cornus officinalis (Cornaceae). The structures of the compounds were identified using physical and spectroscopic methods, as well as by comparison of their data with values reported in the literature. All the isolates were evaluated in vitro for inhibitory activity against the formation of advanced glycation end-products (AGEs) and rat lens aldose reductase (RLAR). Compounds 1-6 were subjected to further bioassay to examine their inhibitory effects on AGE cross-linking. The opacity of lenses was significantly prevented when treated with 3 in an ex vivo experiment.

  9. Scopoletin from the flower buds of Magnolia fargesii inhibits protein glycation, aldose reductase, and cataractogenesis ex vivo.

    PubMed

    Lee, Jun; Kim, Nan Hee; Nam, Joo Won; Lee, Yun Mi; Jang, Dae Sik; Kim, Young Sook; Nam, Sang Hae; Seo, Eun-Kyoung; Yang, Min Suk; Kim, Jin Sook

    2010-09-01

    Five compounds previously known structures, scopoletin (1), northalifoline (2), stigmast-4-en-3-one (3), tiliroside (4), and oplopanone (5) were obtained from the flower buds of Magnolia fargesii using chromatographic separation methods. The structures of 1-5 were identified by the interpretation of their spectroscopic data including 1D- and 2D-NMR as well as by comparison with reported values. Three compounds 1-3 were found from M. fargesii for the first time in this study. All the isolates (1-5) were subjected to in vitro bioassays to evaluate the inhibitory activity on advanced glycation end products formation and rat lens aldose reductase (RLAR). Compound 1 showed a remarkable inhibitory activity on advanced glycation end products formation with IC(50) value of 2.93 μM (aminoguanidine: 961 μM), and showed a significant RLAR inhibitory activity with IC(50) value of 22.5 μM (3.3-tetramethyleneglutaric acid: 28.7 μM). Compound 4 exhibited potent inhibitory activity against RLAR (IC(50) = 14.9 μM). In the further experiment ex vivo, cataractogenesis of rat lenses induced with xylose was significantly inhibited by compound 1 treatment.

  10. Quantum mechanical calculation of electric fields and vibrational Stark shifts at active site of human aldose reductase

    SciTech Connect

    Wang, Xianwei; Zhang, John Z. H.; He, Xiao

    2015-11-14

    Recent advance in biophysics has made it possible to directly measure site-specific electric field at internal sites of proteins using molecular probes with C = O or C≡N groups in the context of vibrational Stark effect. These measurements directly probe changes of electric field at specific protein sites due to, e.g., mutation and are very useful in protein design. Computational simulation of the Stark effect based on force fields such as AMBER and OPLS, while providing good insight, shows large errors in comparison to experimental measurement due to inherent difficulties associated with point charge based representation of force fields. In this study, quantum mechanical calculation of protein’s internal electrostatic properties and vibrational Stark shifts was carried out by using electrostatically embedded generalized molecular fractionation with conjugate caps method. Quantum calculated change of mutation-induced electric field and vibrational Stark shift is reported at the internal probing site of enzyme human aldose reductase. The quantum result is in much better agreement with experimental data than those predicted by force fields, underscoring the deficiency of traditional point charge models describing intra-protein electrostatic properties.

  11. Evaluation of in vitro aldose reductase inhibitory potential of different fraction of Hybanthus enneaspermus Linn F. Muell

    PubMed Central

    Patel, DK; Kumar, R; Kumar, M; Sairam, K; Hemalatha, S

    2012-01-01

    Objective To evaluate the aldose reductase inhibitory (ARI) activity of different fractions of Hybanthus enneaspermus for potential use in diabetic cataract. Methods Total phenol and flavonoid content of different fractions was determined. ARI activity of different fractions in rat lens was investigated in vitro. Results The results showed significant level of phenolic and flavonoid content in ethyl acetate fraction [total phenol (212.15±0.79 mg/g), total flavonoid (39.11±2.27 mg/g)] and aqueous fraction [total phenol (140.62±0.57 mg/g), total flavonoid (26.07±1.49 mg/g)] as compared with the chloroform fraction [total phenol (68.56±0.51 mg/g), total flavonoid (13.41±0.82 mg/g)] and petrolium ether fraction [total phenol (36.68±0.43 mg/g), total flavonoid (11.55±1.06 mg/g)]. There was a significant difference in the ARI activity of each fraction, and it was found to be the highest in ethyl acetate fraction [IC50 (49.26±1.76 µg/mL)] followed by aqueous extract [IC50 (70.83±2.82 µg/mL)] and it was least in the petroleum ether fraction [IC50 (118.89±0.71 µg/mL)]. Chloroform fraction showed moderate activity [IC50 (98.52±1.80 µg/mL)]. Conclusions Different fractions showed significanct amount of ARI activity, where in ethyl acetate fraction it was found to be maximum which may be due to its high phenolic and flavonoid content. The extract after further evaluation may be used in the treatment of diabetic cataract. PMID:23569883

  12. [Role of heat shock proteins, aldose reductase, Bcl-2 protein and microRNA in the mechanism of delayed preconditioning of heart].

    PubMed

    Lishmanov, Iu B; Maslov, L N; Khaliulin, I G; Zhang, Y; Pei, J -M

    2010-05-01

    Analysis of published data allows affirming that heat shock proteins (HSP) play an important role in the mechanism of cardioprotective effect of delayed preconditioning. However, HSP in all probability are non-end effectors but mediators of preconditioning because a peak of their levels in myocardium does not concur with maximal elevation of cardiac tolerance to impact of ischemia and reperfusion. There are bases to think that aldose reductase and Bcl-2 protein are claimants to the role of end-effectors of delayed preconditioning but microRNAs serve as mediators of forming increased cardiac tolerance to ischemia-reperfusion. PMID:20583571

  13. Design of an Amide N-glycoside Derivative of β-Glucogallin: A Stable, Potent, and Specific Inhibitor of Aldose Reductase

    PubMed Central

    Li, Linfeng; Chang, Kun-Che; Zhou, Yaming; Shieh, Biehuoy; Ponder, Jessica; Abraham, Adedoyin D.; Ali, Hadi; Snow, Anson; Petrash, J. Mark; LaBarbera, Daniel V.

    2014-01-01

    β-glucogallin (BGG), a major component of the Emblica officinalis medicinal plant, is a potent and selective inhibitor of aldose-reductase (AKR1B1). New linkages (ether/triazole/amide) were introduced via high yielding, efficient syntheses to replace the labile ester, and an original 2-step (90%) preparation of BGG was developed. Inhibition of AKR1B1was assessed in vitro and using transgenic lens organ cultures, which identified the amide linked glucoside (BGA) as a stable, potent and selective lead therapeutic toward the treatment of diabetic eye disease. PMID:24341381

  14. Gedunin abrogates aldose reductase, PI3K/Akt/mToR, and NF-κB signaling pathways to inhibit angiogenesis in a hamster model of oral carcinogenesis.

    PubMed

    Kishore T, Kranthi Kiran; Ganugula, Raghu; Gade, Deepak Reddy; Reddy, Geereddy Bhanuprakash; Nagini, Siddavaram

    2016-02-01

    Aberrant activation of oncogenic signaling pathways plays a central role in tumor development and progression. The aim of this present study was to investigate the chemopreventive effects of the neem limonoid gedunin in the hamster model of oral cancer based on its ability to modulate aldose reductase (AR), phosphatidyl inositol-3-kinase (PI3K)/Akt, and nuclear factor kappa B (NF-κB) pathways to block angiogenesis. Administration of gedunin suppressed the development of HBP carcinomas by inhibiting PI3K/Akt and NF-κB pathways through the inactivation of Akt and inhibitory kappa B kinase (IKK), respectively. Immunoblot and molecular docking interactions revealed that inhibition of these signaling pathways may be mediated via inactivation of AR by gedunin. Gedunin blocked angiogenesis by downregulating the expression of miR-21 and the pro-angiogenic factors vascular endothelial growth factor and hypoxia inducible factor-1 alpha (HIF-1α). In conclusion, the results of the present study provide compelling evidence that gedunin prevents progression of hamster buccal pouch (HBP) carcinomas via inhibition of the kinases Akt, IKK, and AR, and the oncogenic transcription factors NF-κB and HIF-1α to block angiogenesis. PMID:26342697

  15. Design and synthesis of pyridazinone-substituted benzenesulphonylurea derivatives as anti-hyperglycaemic agents and inhibitors of aldose reductase - an enzyme embroiled in diabetic complications.

    PubMed

    Yaseen, Raed; Pushpalatha, H; Reddy, G Bhanuprakash; Ismael, Ameer; Ahmed, Ayad; Dheyaa, Alhamza; Ovais, Syed; Rathore, Pooja; Samim, Mohammed; Akthar, Mymoona; Sharma, Kalicharan; Shafi, Syed; Singh, Surender; Javed, Kalim

    2016-12-01

    Thirty new aryl-pyridazinone-substituted benzenesulphonylurea derivatives (I-XXX) were synthesized and evaluated for their anti-hyperglycaemic activity in glucose-fed hyperglycaemic normal rats. Twenty-three compounds (III-XI, XIV-XVII, XIX-XXIV, XXVI and XXVIII-XXX) showed more or comparable area under the curve (AUC) reduction percentage (ranging from 21.9% to 35.5%) as compared to the standard drug gliclazide (22.0%). On the basis of docking results, 18 compounds were screened for their in vitro ability to inhibit rat lens aldose reductase. Ten compounds (III-VI, XII, XVI-XVIII, XXI and XXVII) showed ARI activity with IC50 ranging from 34 to 242 μM. Out of these, two compounds IV and V showed best ARI activity which is comparable with that of quercetin. As a result, two compounds (IV and V) possessing significant dual action (anti-hyperglycaemic and aldose reductase inhibition) were identified and may be used as lead compounds for developing new drugs. PMID:26879420

  16. Phenolic Compounds from the Leaves and Twigs of Osteomeles schwerinae That Inhibit Rat Lens Aldose Reductase and Vessel Dilation in Zebrafish Larvae.

    PubMed

    Lee, Ik-Soo; Jung, Seung-Hyun; Lee, Yun Mi; Choi, So-Jin; Sun, Hang; Kim, Jin Sook

    2015-09-25

    Three new phenolic biphenyl derivatives (1-3) and one new lignan glycoside (4) were isolated from the leaves and twigs of Osteomeles schwerinae. The structures of the new compounds were established by spectroscopic data interpretation. The inhibitory effects of 1-4 on rat lens aldose reductase in vitro were examined, and compounds 1-3 markedly inhibited the enzyme with IC50 values of 3.8 to 13.8 μM. In addition, the effects of these isolates on the dilation of hyaloid-retinal vessels induced by high glucose (HG) in zebrafish larvae were investigated. Compound 1 was the most effective in reducing HG-induced dilation of hyaloid-retinal vessels. PMID:26331986

  17. Detoxifying Enzymes at the Cross-Roads of Inflammation, Oxidative Stress, and Drug Hypersensitivity: Role of Glutathione Transferase P1-1 and Aldose Reductase.

    PubMed

    Sánchez-Gómez, Francisco J; Díez-Dacal, Beatriz; García-Martín, Elena; Agúndez, José A G; Pajares, María A; Pérez-Sala, Dolores

    2016-01-01

    Phase I and II enzymes are involved in the metabolism of endogenous reactive compounds as well as xenobiotics, including toxicants and drugs. Genotyping studies have established several drug metabolizing enzymes as markers for risk of drug hypersensitivity. However, other candidates are emerging that are involved in drug metabolism but also in the generation of danger or costimulatory signals. Enzymes such as aldo-keto reductases (AKR) and glutathione transferases (GST) metabolize prostaglandins and reactive aldehydes with proinflammatory activity, as well as drugs and/or their reactive metabolites. In addition, their metabolic activity can have important consequences for the cellular redox status, and impacts the inflammatory response as well as the balance of inflammatory mediators, which can modulate epigenetic factors and cooperate or interfere with drug-adduct formation. These enzymes are, in turn, targets for covalent modification and regulation by oxidative stress, inflammatory mediators, and drugs. Therefore, they constitute a platform for a complex set of interactions involving drug metabolism, protein haptenation, modulation of the inflammatory response, and/or generation of danger signals with implications in drug hypersensitivity reactions. Moreover, increasing evidence supports their involvement in allergic processes. Here, we will focus on GSTP1-1 and aldose reductase (AKR1B1) and provide a perspective for their involvement in drug hypersensitivity. PMID:27540362

  18. Detoxifying Enzymes at the Cross-Roads of Inflammation, Oxidative Stress, and Drug Hypersensitivity: Role of Glutathione Transferase P1-1 and Aldose Reductase

    PubMed Central

    Sánchez-Gómez, Francisco J.; Díez-Dacal, Beatriz; García-Martín, Elena; Agúndez, José A. G.; Pajares, María A.; Pérez-Sala, Dolores

    2016-01-01

    Phase I and II enzymes are involved in the metabolism of endogenous reactive compounds as well as xenobiotics, including toxicants and drugs. Genotyping studies have established several drug metabolizing enzymes as markers for risk of drug hypersensitivity. However, other candidates are emerging that are involved in drug metabolism but also in the generation of danger or costimulatory signals. Enzymes such as aldo-keto reductases (AKR) and glutathione transferases (GST) metabolize prostaglandins and reactive aldehydes with proinflammatory activity, as well as drugs and/or their reactive metabolites. In addition, their metabolic activity can have important consequences for the cellular redox status, and impacts the inflammatory response as well as the balance of inflammatory mediators, which can modulate epigenetic factors and cooperate or interfere with drug-adduct formation. These enzymes are, in turn, targets for covalent modification and regulation by oxidative stress, inflammatory mediators, and drugs. Therefore, they constitute a platform for a complex set of interactions involving drug metabolism, protein haptenation, modulation of the inflammatory response, and/or generation of danger signals with implications in drug hypersensitivity reactions. Moreover, increasing evidence supports their involvement in allergic processes. Here, we will focus on GSTP1-1 and aldose reductase (AKR1B1) and provide a perspective for their involvement in drug hypersensitivity. PMID:27540362

  19. Long-term clinical effects of epalrestat, an aldose reductase inhibitor, on progression of diabetic neuropathy and other microvascular complications: multivariate epidemiological analysis based on patient background factors and severity of diabetic neuropathy

    PubMed Central

    Hotta, N; Kawamori, R; Fukuda, M; Shigeta, Y

    2012-01-01

    Abstract Aims The goal of the study was to evaluate the efficacy of epalrestat, an aldose reductase inhibitor, on diabetic retinopathy and diabetic nephropathy, based on analysis of the results of the Aldose Reductase Inhibitor–Diabetes Complications Trial, a 3-year multicentre comparative clinical trial of conventional therapy (control group) and epalrestat therapy (epalrestat group) in Japanese patients with mild diabetic neuropathy. Methods The subjects of the study were patients enrolled in the Aldose Reductase Inhibitor–Diabetes Complications Trial for whom data for major patient characteristics, severity of diabetic neuropathy at the end of the study and time-courses of diabetic retinopathy and diabetic nephropathy were available (57 and 52 patients from the control and epalrestat groups, respectively). Progression of diabetic retinopathy/nephropathy (a primary endpoint) in relation to major patient characteristics, severity of diabetic neuropathy at the end of the study (assessed from the mean of z-scores in four neurological function tests) and epalrestat treatment were analysed using univariate analysis and multiple logistic regression analysis. Results Progression of diabetic retinopathy/nephropathy was significantly inhibited in the epalrestat group compared with the control group (odds ratio = 0.323, P = 0.014) and was dependent on the severity of diabetic neuropathy at the end of the study (odds ratio = 2.131, P = 0.025). Conclusions Epalrestat prevented progression of diabetic neuropathy and retinopathy/nephropathy. The effect on diabetic retinopathy/nephropathy may have occurred indirectly because of the prevention of progression of diabetic neuropathy, in addition to the inhibitory action of epalrestat on aldose reductase. PMID:22507139

  20. The Prostaglandin F Synthase Activity of the Human Aldose Reductase AKR1B1 Brings New Lenses to Look at Pathologic Conditions

    PubMed Central

    Bresson, Eva; Lacroix-Pépin, Nicolas; Boucher-Kovalik, Sofia; Chapdelaine, Pierre; Fortier, Michel A.

    2012-01-01

    Prostaglandins are important regulators of female reproductive functions to which aldose reductases exhibiting hydroxysteroid dehydrogenase activity also contribute. Our work on the regulation of reproductive function by prostaglandins (PGs), lead us to the discovery that AKR1B5 and later AKR1B1were highly efficient and physiologically relevant PGF synthases. PGE2 and PGF2α are the main prostanoids produced in the human endometrium and proper balance in their relative production is important for normal menstruation and optimal fertility. Recent evidence suggests that PGE2/EP2 and PGF2α/FP may constitute a functional dyad with physiological relevance comparable to the prostacyclin-thromboxane dyad in the vascular system. We have recently reported that AKR1B1 was expressed and modulated in association with PGF2α production in response to IL-1β in the human endometrium. In the present study, we show that the human AKR1B1 (gene ID: 231) also known as ALDR1 or ALR2 is a functional PGF2α synthase in different models of living cells and tissues. Using human endometrial cells, prostate, and vascular smooth muscle cells, cardiomyocytes and endothelial cells we demonstrate that IL-1β is able to up regulate COX-2 and AKR1B1 proteins as well as PGF2α production under normal glucose concentrations. We show that the promoter activity of AKR1B1 gene is increased by IL-1β particularly around the multiple stress response region containing two putative antioxidant response elements adjacent to TonE and AP1. We also show that AKR1B1 is able to regulate PGE2 production through PGF2α acting on its FP receptor and that aldose reductase inhibitors like alrestatin, Statil (ponalrestat), and EBPC exhibit distinct and characteristic inhibition of PGF2α production in different cell models. The PGF synthase activity of AKR1B1 represents a new and important target to regulate ischemic and inflammatory responses associated with several human pathologies. PMID:22654757

  1. Tissue-specific effects of aldose reductase inhibition on fluorescence and cross-linking of extracellular matrix in chronic galactosemia. Relationship to pentosidine cross-links.

    PubMed

    Richard, S; Tamas, C; Sell, D R; Monnier, V M

    1991-08-01

    Chronic experimental hyperglycemia mediated by galactose has been shown to induce browning and cross-linking of rat tail tendon collagen that could be duplicated in vitro by nonenzymatic galactosylation. To investigate the nature of these changes, Sprague-Dawley rats were placed on a 33% galactose diet without and with sorbinil for 6 and 12 mo. Collagen-linked fluorescence and pentosidine cross-links increased with age and galactosemia in tail tendons (P less than 0.001) and skin but were essentially unresponsive to aldose reductase inhibition (ARI). In contrast, tendon breaking time in urea, a likely parameter of cross-linking, was markedly improved (P less than 0.001) by ARI. Fluorescence that was inhibited by sorbinil treatment was increased in pepsin and proteinase K digest of aortic tissue from galactosemic rats (P less than 0.001), but impaired enzymatic digestibility was not observed. Systolic blood pressure as potential consequence of aortic stiffening was not increased in galactosemia. These data suggest that fluorescence in skin and tendon might be in part due to advanced glycosylation and pentosidine formation because these were not decreased by ARI. However, they also suggest that nonfluorescent cross-links may also be forming because, in contrast to fluorescence, tail tendon breaking time was partly corrected by ARI. Thus, it appears that extracellular matrix changes in chronic galactosemia are complex, being partly attributable to advanced glycosylation and partly to polyol-pathway activation.

  2. Pressure-induced inhibition of fast axonal transport of proteins in the rabbit vagus nerve in galactose neuropathy: prevention by an aldose reductase inhibitor.

    PubMed

    McLean, W G

    1988-07-01

    Fast and slow anterograde axonal transport and retrograde axonal transport of proteins were studied in the mainly non-myelinated sensory fibres of the vagus nerve of rabbits fed a diet of 50% galactose over a period of 29 days. Galactose feeding had no effect on the rate or protein composition of slow transport nor on the amount of retrogradely transported proteins. There was a slight retardation of fast transported proteins although their composition was unchanged. The galactose feeding led to a significant increase (p less than 0.005) in nerve water content and nerve galactitol but no significant change in myo-inositol. When 20 mm Hg pressure was applied locally to the cervical vagus nerve, fast transported proteins accumulated proximal to the compression zone in the galactose-fed but not in control rabbits. Administration of the aldose reductase inhibitor Statil (ICI 128436) throughout the experiment prevented the increased susceptibility to pressure and the increase in nerve galactitol and water content. The effects of pressure are similar to those found in the streptozotocin-diabetic rat although the underlying mechanisms may differ.

  3. Effects of Long-Term Treatment with Ranirestat, a Potent Aldose Reductase Inhibitor, on Diabetic Cataract and Neuropathy in Spontaneously Diabetic Torii Rats

    PubMed Central

    Ota, Ayumi; Kakehashi, Akihiro; Toyoda, Fumihiko; Kinoshita, Nozomi; Shinmura, Machiko; Takano, Hiroko; Obata, Hiroto; Matsumoto, Takafumi; Tsuji, Junichi; Dobashi, Yoh; Fujimoto, Wilfred Y.; Kawakami, Masanobu; Kanazawa, Yasunori

    2013-01-01

    We evaluated ranirestat, an aldose reductase inhibitor, in diabetic cataract and neuropathy (DN) in spontaneously diabetic Torii (SDT) rats compared with epalrestat, the positive control. Animals were divided into groups and treated once daily with oral ranirestat (0.1, 1.0, 10 mg/kg) or epalrestat (100 mg/kg) for 40 weeks, normal Sprague-Dawley rats, and untreated SDT rats. Lens opacification was scored from 0 (normal) to 3 (mature cataract). The combined scores (0–6) from both lenses represented the total for each animal. DN was assessed by measuring the motor nerve conduction velocity (MNCV) in the sciatic nerve. Sorbitol and fructose levels were measured in the lens and sciatic nerve 40 weeks after diabetes onset. Cataracts developed more in untreated rats than normal rats (P < 0.01). Ranirestat significantly (P < 0.01) inhibited rapid cataract development; epalrestat did not. Ranirestat significantly reversed the MNCV decrease (40.7 ± 0.6 m/s) in SDT rats dose-dependently (P < 0.01). Epalrestat also reversed the prevented MNCV decrease (P < 0.05). Sorbitol levels in the sciatic nerve increased significantly in SDT rats (2.05 ± 0.10 nmol/g), which ranirestat significantly suppressed dose-dependently, (P < 0.05, <0.01, and <0.01); epalrestat did not. Ranirestat prevents DN and cataract; epalrestat prevents DN only. PMID:23671855

  4. Relation of Na+, K(+)-ATPase to delayed motor nerve conduction velocity: effect of aldose reductase inhibitor, ADN-138, on Na+, K(+)-ATPase activity.

    PubMed

    Hirata, Y; Okada, K

    1990-06-01

    The role of sorbitol, myo-inositol, and Na+, K(+)-adenosine triphosphatase (ATPase) activity on motor nerve conduction velocity (MNCV) in streptozotocin (STZ)-diabetic rats was studied. Reduction of MNCV and Na+, K(+)-ATPase in caudal nerves appeared after 3 weeks of diabetes, and at this time treatment with aldose reductase inhibitor (ARI), ADN-138 and 1% myo-inositol supplement was begun. One percent myo-inositol supplement for 3 weeks resulted in a significant increase in myo-inositol levels in diabetic nerves, but left MNCV and sorbitol levels unchanged. In contrast, treatment with ADN-138 for 3 weeks reduced sorbitol levels in diabetic nerves and resulted in significant increases in MNCV and Na+, K(+)-ATPase in the nerves. Since ADN-138 did not restore myo-inositol levels, the increase in Na+, K(+)-ATPase levels by ADN-138 treatment was independent of myo-inositol levels. Also, nerve Na+ levels in ADN-138-treated rats were reduced and the ratio of K+ to Na+ was raised, while 1% myo-inositol supplement did not affect them. These results suggest that treatment with ADN-138 elevates MNCV through a series of processes: ARI----reduction of sorbitol level----increase in Na+, K(+)-ATPase activity----correction of K+, Na+ imbalance----increase in MNCV.

  5. Triple aldose reductase/α-glucosidase/radical scavenging high-resolution profiling combined with high-performance liquid chromatography-high-resolution mass spectrometry-solid-phase extraction-nuclear magnetic resonance spectroscopy for identification of antidiabetic constituents in crude extract of Radix Scutellariae.

    PubMed

    Tahtah, Yousof; Kongstad, Kenneth T; Wubshet, Sileshi G; Nyberg, Nils T; Jønsson, Louise H; Jäger, Anna K; Qinglei, Sun; Staerk, Dan

    2015-08-21

    In this work, development of a new microplate-based high-resolution profiling assay using recombinant human aldose reductase is presented. Used together with high-resolution radical scavenging and high-resolution α-glucosidase assays, it provided the first report of a triple aldose reductase/α-glucosidase/radical scavenging high-resolution inhibition profile - allowing proof of concept with Radix Scutellariae crude extract as a polypharmacological herbal drug. The triple bioactivity high-resolution profiles were used to pinpoint bioactive compounds, and subsequent structure elucidation was performed with hyphenated high-performance liquid chromatography-high-resolution mass spectrometry-solid-phase extraction-nuclear magnetic resonance spectroscopy. The only α-glucosidase inhibitor was baicalein, whereas main aldose reductase inhibitors in the crude extract were baicalein and skullcapflavone II, and main radical scavengers were ganhuangemin, viscidulin III, baicalin, oroxylin A 7-O-glucuronide, wogonoside, baicalein, wogonin, and skullcapflavone II.

  6. Diabetic neuropathy in db/db mice develops independently of changes in ATPase and aldose reductase. A biochemical and immunohistochemical study.

    PubMed

    Bianchi, R; Marelli, C; Marini, P; Fabris, M; Triban, C; Fiori, M G

    1990-03-01

    ATPase activity was investigated in sciatic and optic nerves of female mutant diabetic C57Bl/Ks (db/db) mice and age-matched control mice (db/m and m/m). Nerves from animals aged 50, 70, 125, 180 and 280 days were assayed in vitro for ATPase activity in the presence or absence of ouabain: the ouabain-sensitive fraction contained Na+,K(+)-ATPase. Enzymatic activity was compared within and between age-matched groups. No significant difference in Na+,K(+)-ATPase activity was detected between the diabetic and control mice, whether expressed as mumol Pi/h-1 formed per gramme wet weight or per nerve (protein content). The activity decreased by about 25% in both the sciatic and optic nerves of the oldest animals. These results were strikingly similar in all groups, regardless of the type of nerve examined, confirming that the development of neuropathy in this animal model is unrelated to the postulated derangement of Na+,K(+)-ATPase activity. Among possible explanations, a lack of polyol pathway activation was investigated by staining the sciatic nerves of animals from all groups with the peroxidase-antiperoxidase procedure using a polyclonal antiserum raised against the enzyme aldose reductase. Histological sections of all nerves were consistently negative, suggesting that these animals actually lack the enzyme involved in activating the self-perpetuating metabolic cycle leading to deranged nerve function. The db/db mouse appears to present particular biochemical changes which merit attention with a view to clarifying the pathogenesis of diabetic neuropathy.

  7. Effects of the New Aldose Reductase Inhibitor Benzofuroxane Derivative BF-5m on High Glucose Induced Prolongation of Cardiac QT Interval and Increase of Coronary Perfusion Pressure

    PubMed Central

    Di Filippo, C.; Ferraro, B.; Maisto, R.; Trotta, M. C.; Di Carluccio, N.; Sartini, S.; La Motta, C.; Ferraraccio, F.; Rossi, F.; D'Amico, M.

    2016-01-01

    This study investigated the effects of the new aldose reductase inhibitor benzofuroxane derivative 5(6)-(benzo[d]thiazol-2-ylmethoxy)benzofuroxane (BF-5m) on the prolongation of cardiac QT interval and increase of coronary perfusion pressure (CPP) in isolated, high glucose (33.3 mM D-glucose) perfused rat hearts. BF-5m was dissolved in the Krebs solution at a final concentration of 0.01 μM, 0.05 μM, and 0.1 μM. 33.3 mM D-glucose caused a prolongation of the QT interval and increase of CPP up to values of 190 ± 12 ms and 110 ± 8 mmHg with respect to the values of hearts perfused with standard Krebs solution (11.1 mM D-glucose). The QT prolongation was reduced by 10%, 32%, and 41%, respectively, for the concentration of BF-5m 0.01 μM, 0.05 μM, and 0.1 μM. Similarly, the CPP was reduced by 20% for BF-5m 0.05 μM and by 32% for BF-5m 0.1 μM. BF-5m also increased the expression levels of sirtuin 1, MnSOD, eNOS, and FOXO-1, into the heart. The beneficial actions of BF-5m were partly abolished by the pretreatment of the rats with the inhibitor of the sirtuin 1 activity EX527 (10 mg/kg/day/7 days i.p.) prior to perfusion of the hearts with high glucose + BF-5m (0.1 μM). Therefore, BF-5m supplies cardioprotection from the high glucose induced QT prolongation and increase of CPP. PMID:26839893

  8. A meta-analysis of trials on aldose reductase inhibitors in diabetic peripheral neuropathy. The Italian Study Group. The St. Vincent Declaration.

    PubMed

    Nicolucci, A; Carinci, F; Cavaliere, D; Scorpiglione, N; Belfiglio, M; Labbrozzi, D; Mari, E; Benedetti, M M; Tognoni, G; Liberati, A

    1996-12-01

    Peripheral neuropathy is one of the most common and disabling long-term sequelae of diabetes mellitus. Aldose reductase inhibitors (ARIs) have been proposed and are increasingly used in many countries for the prevention and treatment of diabetic neuropathy. The aim of this study was to review existing evidence on the effectiveness of ARIs in the treatment of peripheral diabetic neuropathy, with particular reference to the type and clinical relevance of the end point used and to the consistency of results across studies. Thirteen randomized clinical trials (RTCs) comparing ARIs with placebo, published between 1981 and 1993 were included in the meta-analysis. Nerve conduction velocity (NCV) was the only end point reported in all trials. Treatment effect was thus evaluated in terms of NCV mean difference in four different nerves: median motor, median sensory, peroneal motor, and sural sensory. A statistically significant reduction in decline of median motor NCV was present in the treated group as compared to the control group (mean 0.91 ms-1; 95% CI 0.41-1.42 ms-1). For peroneal motor, median sensory, and sural sensory nerves results did not show any clear benefit for patients treated with ARIs. When the analysis was limited to trials with at least 1-year treatment duration, a significant effect was present for peroneal motor NCV (mean 1.24 ms-1; 95% CI 0.32-2.15 ms-1) and a benefit of borderline statistical significance was also present for median motor NCV (mean 0.69 ms-1; 95% CI-0.07-1.45 ms-1). A heterogeneous picture emerged when looking at the results of different studies and serious inconsistencies were also present in the direction of treatment effects among nerves in the same studies. Although the results of 1-year treatment on motor NCV seem encouraging, the uncertainty about the reliability of the end-point employed and the short treatment duration do not allow any clear conclusion about the efficacy of ARIs in the treatment of peripheral diabetic

  9. Functional genomic studies of aldo-keto reductases.

    PubMed

    Petrash, J M; Murthy, B S; Young, M; Morris, K; Rikimaru, L; Griest, T A; Harter, T

    2001-01-30

    Aldose reductase (AR) is considered a potential mediator of diabetic complications and is a drug target for inhibitors of diabetic retinopathy and neuropathy in clinical trials. However, the physiological role of this enzyme still has not been established. Since effective inhibition of diabetic complications will require early intervention, it is important to delineate whether AR fulfills a physiological role that cannot be compensated by an alternate aldo-keto reductase. Functional genomics provides a variety of powerful new tools to probe the physiological roles of individual genes, especially those comprising gene families. Several eucaryotic genomes have been sequenced and annotated, including yeast, nematode and fly. To probe the function of AR, we have chosen to utilize the budding yeast Saccharomyces cerevisiae as a potential model system. Unlike Caenorhabditis elegans and D. melanogaster, yeast provides a more desirable system for our studies because its genome is manipulated more readily and is able to sustain multiple gene deletions in the presence of either drug or auxotrophic selectable markers. Using BLAST searches against the human AR gene sequence, we identified six genes in the complete S. cerevisiae genome with strong homology to AR. In all cases, amino acids thought to play important catalytic roles in human AR are conserved in the yeast AR-like genes. All six yeast AR-like open reading frames (ORFs) have been cloned into plasmid expression vectors. Substrate and AR inhibitor specificities have been surveyed on four of the enzyme forms to identify, which are the most functionally similar to human AR. Our data reveal that two of the enzymes (YDR368Wp and YHR104Wp) are notable for their similarity to human AR in terms of activity with aldoses and substituted aromatic aldehydes. Ongoing studies are aimed at characterizing the phenotypes of yeast strains containing single and multiple knockouts of the AR-like genes. PMID:11306085

  10. Characterization, real-time quantification and in silico modeling of arsenate reductase (arsC) genes in arsenic-resistant Herbaspirillum sp. GW103.

    PubMed

    Govarthanan, Muthusamy; Lee, Sang-Myeong; Kamala-Kannan, Seralathan; Oh, Byung-Taek

    2015-04-01

    This study investigated the mechanism of arsenic resistance in the diazotrophic bacterium Herbaspirillum sp. GW103 isolated from rhizosphere soil of Phragmites austrails. The isolate Herbaspirillum sp. GW103 exhibited maximum tolerance to arsenic (550 mg/L). Four different arsenate reductase (arsC) genes (arsC1, arsC2, arsC3 and arsC4) were located in the genome of the isolate Herbaspirillum sp. GW103. The expression pattern of the arsC1 differed from other genes. All four types of arsC genes had different protein secondary structures and stereochemical properties. Molecular modeling and structural analysis of arsC genes revealed close structural homology with arsC family proteins from Escherichia coli (PDB ID: 1I9D) and Pseudomonas aeruginosa (PDB ID: 1RW1). PMID:25744778

  11. Effect of a new aldose reductase inhibitor, 8'-chloro-2',3'-dihydrospiro [pyrrolidine-3,6'(5'H)-pyrrolo[1,2,3-de] [1,4]benzoxazine]-2,5,5'- trione (ADN-138), on delayed motor nerve conduction velocity in streptozotocin-diabetic rats.

    PubMed

    Hirata, Y; Fujimori, S; Okada, K

    1988-02-01

    The effects of a chemically new type of aldose reductase inhibitor, ADN-138, on delayed motor nerve conduction velocity (MNCV) and sciatic nerve sorbitol, fructose and myo-inositol levels were studied in streptozotocin-diabetic rats. MNCV in rats was significantly delayed after 3 weeks of diabetes and ADN-138 treatment was started at this point. Treatment of diabetics with ADN-138 at 5 and 20 but not 1 mg/kg/d for 3 weeks resulted in a significant increase in MNCV and reduced sorbitol levels to or below those of nondiabetic controls. However, fructose, though decreased in a dose-dependent manner, was not normalized. The reference drug, Sorbinil, showed similar effects on them. After the 3 weeks of ADN-138(20 mg/kg/d) treatment, diabetics were left on ADN-138 or continued further to be treated with it for 3 weeks. The withdrawal of ADN-138 prevented a further increase in MNCV and restored sorbitol and fructose to nontreated diabetic levels, and myo-inositol levels declined. In contrast, the ADN-138-continued group kept improving its MNCV and normalized sorbitol and myo-inositol. These results suggest that polyol accumulation is responsible for delayed MNCV and that the action of ADN-138 on MNCV reflected reversibility of metabolic function in diabetics.

  12. Structure and function of Caulobacter crescentus aldose-aldose oxidoreductase.

    PubMed

    Taberman, Helena; Andberg, Martina; Koivula, Anu; Hakulinen, Nina; Penttilä, Merja; Rouvinen, Juha; Parkkinen, Tarja

    2015-12-15

    Aldose-aldose oxidoreductase (Cc AAOR) is a recently characterized enzyme from the bacterial strain Caulobacter crescentus CB15 belonging to the glucose-fructose oxidoreductase/inositol dehydrogenase/rhizopine catabolism protein (Gfo/Idh/MocA) family. Cc AAOR catalyses the oxidation and reduction of a panel of aldose monosaccharides using a tightly bound NADP(H) cofactor that is regenerated in the catalytic cycle. Furthermore, Cc AAOR can also oxidize 1,4-linked oligosaccharides. In the present study, we present novel crystal structures of the dimeric Cc AAOR in complex with the cofactor and glycerol, D-xylose, D-glucose, maltotriose and D-sorbitol determined to resolutions of 2.0, 1.8, 1.7, 1.9 and 1.8 Å (1 Å=0.1 nm), respectively. These complex structures allowed for a detailed analysis of the ligand-binding interactions. The structures showed that the C1 carbon of a substrate, which is either reduced or oxidized, is close to the reactive C4 carbon of the nicotinamide ring of NADP(H). In addition, the O1 hydroxy group of the substrate, which is either protonated or deprotonated, is unexpectedly close to both Lys(104) and Tyr(189), which may both act as a proton donor or acceptor. This led us to hypothesize that this intriguing feature could be beneficial for Cc AAOR to catalyse the reduction of a linear form of a monosaccharide substrate and the oxidation of a pyranose form of the same substrate in a reaction cycle, during which the bound cofactor is regenerated.

  13. Effect of thermal stability on protein adsorption to silica using homologous aldo-keto reductases

    PubMed Central

    Felsovalyi, Flora; Patel, Tushar; Mangiagalli, Paolo; Kumar, Sanat K; Banta, Scott

    2012-01-01

    Gaining more insight into the mechanisms governing the behavior of proteins at solid/liquid interfaces is particularly relevant in the interaction of high-value biologics with storage and delivery device surfaces, where adsorption-induced conformational changes may dramatically affect biocompatibility. The impact of structural stability on interfacial behavior has been previously investigated by engineering nonwild-type stability mutants. Potential shortcomings of such approaches include only modest changes in thermostability, and the introduction of changes in the topology of the proteins when disulfide bonds are incorporated. Here we employ two members of the aldo-keto reductase superfamily (alcohol dehydrogenase, AdhD and human aldose reductase, hAR) to gain a new perspective on the role of naturally occurring thermostability on adsorbed protein arrangement and its subsequent impact on desorption. Unexpectedly, we find that during initial adsorption events, both proteins have similar affinity to the substrate and undergo nearly identical levels of structural perturbation. Interesting differences between AdhD and hAR occur during desorption and both proteins exhibit some level of activity loss and irreversible conformational change upon desorption. Although such surface-induced denaturation is expected for the less stable hAR, it is remarkable that the extremely thermostable AdhD is similarly affected by adsorption-induced events. These results question the role of thermal stability as a predictor of protein adsorption/desorption behavior. PMID:22619179

  14. Aldo-Keto Reductases 1B in Endocrinology and Metabolism

    PubMed Central

    Pastel, Emilie; Pointud, Jean-Christophe; Volat, Fanny; Martinez, Antoine; Lefrançois-Martinez, Anne-Marie

    2012-01-01

    The aldose reductase (AR; human AKR1B1/mouse Akr1b3) has been the focus of many research because of its role in diabetic complications. The starting point of these alterations is the massive entry of glucose in polyol pathway where it is converted into sorbitol by this enzyme. However, the issue of AR function in non-diabetic condition remains unresolved. AR-like enzymes (AKR1B10, Akr1b7, and Akr1b8) are highly related isoforms often co-expressed with bona fide AR, making functional analysis of one or the other isoform a challenging task. AKR1B/Akr1b members share at least 65% protein identity and the general ability to reduce many redundant substrates such as aldehydes provided from lipid peroxidation, steroids and their by-products, and xenobiotics in vitro. Based on these properties, AKR1B/Akr1b are generally considered as detoxifying enzymes. Considering that divergences should be more informative than similarities to help understanding their physiological functions, we chose to review specific hallmarks of each human/mouse isoforms by focusing on tissue distribution and specific mechanisms of gene regulation. Indeed, although the AR shows ubiquitous expression, AR-like proteins exhibit tissue-specific patterns of expression. We focused on three organs where certain isoforms are enriched, the adrenal gland, enterohepatic, and adipose tissues and tried to connect recent enzymatic and regulation data with endocrine and metabolic functions of these organs. We presented recent mouse models showing unsuspected physiological functions in the regulation of glucido-lipidic metabolism and adipose tissue homeostasis. Beyond the widely accepted idea that AKR1B/Akr1b are detoxification enzymes, these recent reports provide growing evidences that they are able to modify or generate signal molecules. This conceptually shifts this class of enzymes from unenviable status of scavenger to upper class of messengers. PMID:22876234

  15. Aldo-Keto Reductases 1B in Endocrinology and Metabolism.

    PubMed

    Pastel, Emilie; Pointud, Jean-Christophe; Volat, Fanny; Martinez, Antoine; Lefrançois-Martinez, Anne-Marie

    2012-01-01

    The aldose reductase (AR; human AKR1B1/mouse Akr1b3) has been the focus of many research because of its role in diabetic complications. The starting point of these alterations is the massive entry of glucose in polyol pathway where it is converted into sorbitol by this enzyme. However, the issue of AR function in non-diabetic condition remains unresolved. AR-like enzymes (AKR1B10, Akr1b7, and Akr1b8) are highly related isoforms often co-expressed with bona fide AR, making functional analysis of one or the other isoform a challenging task. AKR1B/Akr1b members share at least 65% protein identity and the general ability to reduce many redundant substrates such as aldehydes provided from lipid peroxidation, steroids and their by-products, and xenobiotics in vitro. Based on these properties, AKR1B/Akr1b are generally considered as detoxifying enzymes. Considering that divergences should be more informative than similarities to help understanding their physiological functions, we chose to review specific hallmarks of each human/mouse isoforms by focusing on tissue distribution and specific mechanisms of gene regulation. Indeed, although the AR shows ubiquitous expression, AR-like proteins exhibit tissue-specific patterns of expression. We focused on three organs where certain isoforms are enriched, the adrenal gland, enterohepatic, and adipose tissues and tried to connect recent enzymatic and regulation data with endocrine and metabolic functions of these organs. We presented recent mouse models showing unsuspected physiological functions in the regulation of glucido-lipidic metabolism and adipose tissue homeostasis. Beyond the widely accepted idea that AKR1B/Akr1b are detoxification enzymes, these recent reports provide growing evidences that they are able to modify or generate signal molecules. This conceptually shifts this class of enzymes from unenviable status of scavenger to upper class of messengers.

  16. Functional studies of aldo-keto reductases in Saccharomyces cerevisiae.

    PubMed

    Chang, Qing; Griest, Terry A; Harter, Theresa M; Petrash, J Mark

    2007-03-01

    We utilized the budding yeast Saccharomyces cerevisiae as a model to systematically explore physiological roles for yeast and mammalian aldo-keto reductases. Six open reading frames encoding putative aldo-keto reductases were identified when the yeast genome was queried against the sequence for human aldose reductase, the prototypical mammalian aldo-keto reductase. Recombinant proteins produced from five of these yeast open reading frames demonstrated NADPH-dependent reductase activity with a variety of aldehyde and ketone substrates. A triple aldo-keto reductase null mutant strain demonstrated a glucose-dependent heat shock phenotype which could be rescued by ectopic expression of human aldose reductase. Catalytically-inactive mutants of human or yeast aldo-keto reductases failed to effect a rescue of the heat shock phenotype, suggesting that the phenotype results from either an accumulation of one or more unmetabolized aldo-keto reductase substrates or a synthetic deficiency of aldo-keto reductase products generated in response to heat shock stress. These results suggest that multiple aldo-keto reductases fulfill functionally redundant roles in the stress response in yeast. PMID:17140678

  17. Functional studies of aldo-keto reductases in Saccharomyces cerevisiae*

    PubMed Central

    Chang, Qing; Griest, Terry A.; Harter, Theresa M.; Petrash, J. Mark

    2007-01-01

    SUMMARY We utilized the budding yeast Saccharomyces cerevisiae as a model to systematically explore physiological roles for yeast and mammalian aldo-keto reductases. Six open reading frames encoding putative aldo-keto reductases were identified when the yeast genome was queried against the sequence for human aldose reductase, the prototypical mammalian aldo-keto reductase. Recombinant proteins produced from five of these yeast open reading frames demonstrated NADPH-dependent reductase activity with a variety of aldehyde and ketone substrates. A triple aldo-keto reductase null mutant strain demonstrated a glucose-dependent heat shock phenotype which could be rescued by ectopic expression of human aldose reductase. Catalytically-inactive mutants of human or yeast aldo-keto reductases failed to effect a rescue of the heat shock phenotype, suggesting that the phenotype results from either an accumulation of one or more unmetabolized aldo-keto reductase substrates or a synthetic deficiency of aldo-keto reductase products generated in response to heat shock stress. These results suggest that multiple aldo-keto reductases fulfill functionally redundant roles in the stress response in yeast. PMID:17140678

  18. Extreme ultraviolet photoionization of aldoses and ketoses

    NASA Astrophysics Data System (ADS)

    Shin, Joong-Won; Dong, Feng; Grisham, Michael E.; Rocca, Jorge J.; Bernstein, Elliot R.

    2011-04-01

    Gas phase monosaccharides (2-deoxyribose, ribose, arabinose, xylose, lyxose, glucose galactose, fructose, and tagatose), generated by laser desorption of solid sample pellets, are ionized with extreme ultraviolet photons (EUV, 46.9 nm, 26.44 eV). The resulting fragment ions are analyzed using a time of flight mass spectrometer. All aldoses yield identical fragment ions regardless of size, and ketoses, while also generating same ions as aldoses, yields additional features. Extensive fragmentation of the monosaccharides is the result the EUV photons ionizing various inner valence orbitals. The observed fragmentation patterns are not dependent upon hydrogen bonding structure or OH group orientation.

  19. Catalytic anomeric aminoalkynylation of unprotected aldoses.

    PubMed

    Kimura, Yasuaki; Ito, Soichi; Shimizu, Yohei; Kanai, Motomu

    2013-08-16

    A copper(I)-catalyzed anomeric aminoalkynylation reaction of unprotected aldoses was realized. Use of an electron-deficient phosphine ligand, boric acid to stabilize the iminium intermediate, and a protic additive (IPA) to presumably enhance reversible carbohydrate-boron complexation were all essential for efficient conversion. The reaction proceeded well even with a natural disaccharide substrate, suggesting that the developed catalytic reaction could be useful for the synthesis of glycoconjugates with minimum use of protecting groups. PMID:23901780

  20. The aldo-keto reductase superfamily homepage.

    PubMed

    Hyndman, David; Bauman, David R; Heredia, Vladi V; Penning, Trevor M

    2003-02-01

    The aldo-keto reductases (AKRs) are one of the three enzyme superfamilies that perform oxidoreduction on a wide variety of natural and foreign substrates. A systematic nomenclature for the AKR superfamily was adopted in 1996 and was updated in September 2000 (visit www.med.upenn.edu/akr). Investigators have been diligent in submitting sequences of functional proteins to the Web site. With the new additions, the superfamily contains 114 proteins expressed in prokaryotes and eukaryotes that are distributed over 14 families (AKR1-AKR14). The AKR1 family contains the aldose reductases, the aldehyde reductases, the hydroxysteroid dehydrogenases and steroid 5beta-reductases, and is the largest. Other families of interest include AKR6, which includes potassium channel beta-subunits, and AKR7 the aflatoxin aldehyde reductases. Two new families include AKR13 (yeast aldose reductase) and AKR14 (Escherichia coli aldehyde reductase). Crystal structures of many AKRs and their complexes with ligands are available in the PDB and accessible through the Web site. Each structure has the characteristic (alpha/beta)(8)-barrel motif of the superfamily, a conserved cofactor binding site and a catalytic tetrad, and variable loop structures that define substrate specificity. Although the majority of AKRs are monomeric proteins of about 320 amino acids in length, the AKR2, AKR6 and AKR7 family may form multimers. To expand the nomenclature to accommodate multimers, we recommend that the composition and stoichiometry be listed. For example, AKR7A1:AKR7A4 (1:3) would designate a tetramer of the composition indicated. The current nomenclature is recognized by the Human Genome Project (HUGO) and the Web site provides a link to genomic information including chromosomal localization, gene boundaries, human ESTs and SNPs and much more.

  1. Characterization of a unique Caulobacter crescentus aldose-aldose oxidoreductase having dual activities.

    PubMed

    Andberg, Martina; Maaheimo, Hannu; Kumpula, Esa-Pekka; Boer, Harry; Toivari, Mervi; Penttilä, Merja; Koivula, Anu

    2016-01-01

    We describe here the characterization of a novel enzyme called aldose-aldose oxidoreductase (Cc AAOR; EC 1.1.99) from Caulobacter crescentus. The Cc AAOR exists in solution as a dimer, belongs to the Gfo/Idh/MocA family and shows homology with the glucose-fructose oxidoreductase from Zymomonas mobilis. However, unlike other known members of this protein family, Cc AAOR is specific for aldose sugars and can be in the same catalytic cycle both oxidise and reduce a panel of monosaccharides at the C1 position, producing in each case the corresponding aldonolactone and alditol, respectively. Cc AAOR contains a tightly-bound nicotinamide cofactor, which is regenerated in this oxidation-reduction cycle. The highest oxidation activity was detected on D-glucose but significant activity was also observed on D-xylose, L-arabinose and D-galactose, revealing that both hexose and pentose sugars are accepted as substrates by Cc AAOR. The configuration at the C2 and C3 positions of the saccharides was shown to be especially important for the substrate binding. Interestingly, besides monosaccharides, Cc AAOR can also oxidise a range of 1,4-linked oligosaccharides having aldose unit at the reducing end, such as lactose, malto- and cello-oligosaccharides as well as xylotetraose. (1)H NMR used to monitor the oxidation and reduction reaction simultaneously, demonstrated that although D-glucose has the highest affinity and is also oxidised most efficiently by Cc AAOR, the reduction of D-glucose is clearly not as efficient. For the overall reaction catalysed by Cc AAOR, the L-arabinose, D-xylose and D-galactose were the most potent substrates.

  2. Substrate specificity and catalytic efficiency of aldo-keto reductases with phospholipid aldehydes.

    PubMed

    Spite, Matthew; Baba, Shahid P; Ahmed, Yonis; Barski, Oleg A; Nijhawan, Kanchan; Petrash, J Mark; Bhatnagar, Aruni; Srivastava, Sanjay

    2007-07-01

    Phospholipid oxidation generates several bioactive aldehydes that remain esterified to the glycerol backbone ('core' aldehydes). These aldehydes induce endothelial cells to produce monocyte chemotactic factors and enhance monocyte-endothelium adhesion. They also serve as ligands of scavenger receptors for the uptake of oxidized lipoproteins or apoptotic cells. The biochemical pathways involved in phospholipid aldehyde metabolism, however, remain largely unknown. In the present study, we have examined the efficacy of the three mammalian AKR (aldo-keto reductase) families in catalysing the reduction of phospholipid aldehydes. The model phospholipid aldehyde POVPC [1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine] was efficiently reduced by members of the AKR1, but not by the AKR6 or the ARK7 family. In the AKR1 family, POVPC reductase activity was limited to AKR1A and B. No significant activity was observed with AKR1C enzymes. Among the active proteins, human AR (aldose reductase) (AKR1B1) showed the highest catalytic activity. The catalytic efficiency of human small intestinal AR (AKR1B10) was comparable with the murine AKR1B proteins 1B3 and 1B8. Among the murine proteins AKR1A4 and AKR1B7 showed appreciably lower catalytic activity as compared with 1B3 and 1B8. The human AKRs, 1B1 and 1B10, and the murine proteins, 1B3 and 1B8, also reduced C-7 and C-9 sn-2 aldehydes as well as POVPE [1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphoethanolamine]. AKR1A4, B1, B7 and B8 catalysed the reduction of aldehydes generated in oxidized C(16:0-20:4) phosphatidylcholine with acyl, plasmenyl or alkyl linkage at the sn-1 position or C(16:0-20:4) phosphatidylglycerol or phosphatidic acid. AKR1B1 displayed the highest activity with phosphatidic acids; AKR1A4 was more efficient with long-chain aldehydes such as 5-hydroxy-8-oxo-6-octenoyl derivatives, whereas AKR1B8 preferred phosphatidylglycerol. These results suggest that proteins of the AKR1A and B families are

  3. Catalytic Isomerization of Biomass‐Derived Aldoses: A Review

    PubMed Central

    Delidovich, Irina

    2016-01-01

    Abstract Selected aldohexoses (d‐glucose, d‐mannose, and d‐galactose) and aldopentoses (d‐xylose, l‐arabinose, and d‐ribose) are readily available components of biopolymers. Isomerization reactions of these substances are very attractive as carbon‐efficient processes to broaden the portfolio of abundant monosaccharides. This review focuses on the chemocatalytic isomerization of aldoses into the corresponding ketoses as well as epimerization of aldoses at C2. Recent advances in the fields of catalysis by bases and Lewis acids are considered. The emphasis is laid on newly uncovered catalytic systems and mechanisms of carbohydrate transformations. PMID:26948404

  4. Catalytic Isomerization of Biomass-Derived Aldoses: A Review.

    PubMed

    Delidovich, Irina; Palkovits, Regina

    2016-03-21

    Selected aldohexoses (D-glucose, D-mannose, and D-galactose) and aldopentoses (D-xylose, L-arabinose, and D-ribose) are readily available components of biopolymers. Isomerization reactions of these substances are very attractive as carbon-efficient processes to broaden the portfolio of abundant monosaccharides. This review focuses on the chemocatalytic isomerization of aldoses into the corresponding ketoses as well as epimerization of aldoses at C2. Recent advances in the fields of catalysis by bases and Lewis acids are considered. The emphasis is laid on newly uncovered catalytic systems and mechanisms of carbohydrate transformations.

  5. Dehydration of different ketoses and aldoses to 5-hydroxymethylfurfural.

    PubMed

    van Putten, Robert-Jan; Soetedjo, Jenny N M; Pidko, Evgeny A; van der Waal, Jan C; Hensen, Emiel J M; de Jong, Ed; Heeres, Hero J

    2013-09-01

    5-Hydroxymethylfurfural (HMF) is considered an important building block for future bio-based chemicals. Here, we present an experimental study using different ketoses (fructose, sorbose, tagatose) and aldoses (glucose, mannose, galactose) under aqueous acidic conditions (65 g L(-1) substrate, 100-160 °C, 33-300 mM H2 SO4 ) to gain insights into reaction pathways for hexose dehydration to HMF. Both reaction rates and HMF selectivities were significantly higher for ketoses than for aldoses, which is in line with literature. Screening and kinetic experiments showed that the reactivity of the different ketoses is a function of the hydroxyl group orientation at the C3 and C4 positions. These results, in combination with DFT calculations, point to a dehydration mechanism involving cyclic intermediates. For aldoses, no influence of the hydroxyl group orientation was observed, indicating a different rate-determining step. The combination of the knowledge from the literature and the findings in this work indicates that aldoses require an isomerization to ketose prior to dehydration to obtain high HMF yields. PMID:24039165

  6. Aldo-Keto Reductases 1B in Adrenal Cortex Physiology.

    PubMed

    Pastel, Emilie; Pointud, Jean-Christophe; Martinez, Antoine; Lefrançois-Martinez, A Marie

    2016-01-01

    Aldose reductase (AKR1B) proteins are monomeric enzymes, belonging to the aldo-keto reductase (AKR) superfamily. They perform oxidoreduction of carbonyl groups from a wide variety of substrates, such as aliphatic and aromatic aldehydes or ketones. Due to the involvement of human aldose reductases in pathologies, such as diabetic complications and cancer, AKR1B subgroup enzymatic properties have been extensively characterized. However, the issue of AKR1B function in non-pathologic conditions remains poorly resolved. Adrenal activities generated large amount of harmful aldehydes from lipid peroxidation and steroidogenesis, including 4-hydroxynonenal (4-HNE) and isocaproaldehyde (4-methylpentanal), which can both be reduced by AKR1B proteins. More recently, some AKR1B isoforms have been shown to be endowed with prostaglandin F synthase (PGFS) activity, suggesting that, in addition to possible scavenger function, they could instigate paracrine signals. Interestingly, the adrenal gland is one of the major sites for human and murine AKR1B expression, suggesting that their detoxifying/signaling activity could be specifically required for the correct handling of adrenal function. Moreover, chronic effects of ACTH result in a coordinated regulation of genes encoding the steroidogenic enzymes and some AKR1B isoforms. This review presents the molecular mechanisms accounting for the adrenal-specific expression of some AKR1B genes. Using data from recent mouse genetic models, we will try to connect their enzymatic properties and regulation with adrenal functions.

  7. Aldo-Keto Reductases 1B in Adrenal Cortex Physiology

    PubMed Central

    Pastel, Emilie; Pointud, Jean-Christophe; Martinez, Antoine; Lefrançois-Martinez, A. Marie

    2016-01-01

    Aldose reductase (AKR1B) proteins are monomeric enzymes, belonging to the aldo-keto reductase (AKR) superfamily. They perform oxidoreduction of carbonyl groups from a wide variety of substrates, such as aliphatic and aromatic aldehydes or ketones. Due to the involvement of human aldose reductases in pathologies, such as diabetic complications and cancer, AKR1B subgroup enzymatic properties have been extensively characterized. However, the issue of AKR1B function in non-pathologic conditions remains poorly resolved. Adrenal activities generated large amount of harmful aldehydes from lipid peroxidation and steroidogenesis, including 4-hydroxynonenal (4-HNE) and isocaproaldehyde (4-methylpentanal), which can both be reduced by AKR1B proteins. More recently, some AKR1B isoforms have been shown to be endowed with prostaglandin F synthase (PGFS) activity, suggesting that, in addition to possible scavenger function, they could instigate paracrine signals. Interestingly, the adrenal gland is one of the major sites for human and murine AKR1B expression, suggesting that their detoxifying/signaling activity could be specifically required for the correct handling of adrenal function. Moreover, chronic effects of ACTH result in a coordinated regulation of genes encoding the steroidogenic enzymes and some AKR1B isoforms. This review presents the molecular mechanisms accounting for the adrenal-specific expression of some AKR1B genes. Using data from recent mouse genetic models, we will try to connect their enzymatic properties and regulation with adrenal functions. PMID:27499746

  8. Aldo-Keto Reductases 1B in Adrenal Cortex Physiology.

    PubMed

    Pastel, Emilie; Pointud, Jean-Christophe; Martinez, Antoine; Lefrançois-Martinez, A Marie

    2016-01-01

    Aldose reductase (AKR1B) proteins are monomeric enzymes, belonging to the aldo-keto reductase (AKR) superfamily. They perform oxidoreduction of carbonyl groups from a wide variety of substrates, such as aliphatic and aromatic aldehydes or ketones. Due to the involvement of human aldose reductases in pathologies, such as diabetic complications and cancer, AKR1B subgroup enzymatic properties have been extensively characterized. However, the issue of AKR1B function in non-pathologic conditions remains poorly resolved. Adrenal activities generated large amount of harmful aldehydes from lipid peroxidation and steroidogenesis, including 4-hydroxynonenal (4-HNE) and isocaproaldehyde (4-methylpentanal), which can both be reduced by AKR1B proteins. More recently, some AKR1B isoforms have been shown to be endowed with prostaglandin F synthase (PGFS) activity, suggesting that, in addition to possible scavenger function, they could instigate paracrine signals. Interestingly, the adrenal gland is one of the major sites for human and murine AKR1B expression, suggesting that their detoxifying/signaling activity could be specifically required for the correct handling of adrenal function. Moreover, chronic effects of ACTH result in a coordinated regulation of genes encoding the steroidogenic enzymes and some AKR1B isoforms. This review presents the molecular mechanisms accounting for the adrenal-specific expression of some AKR1B genes. Using data from recent mouse genetic models, we will try to connect their enzymatic properties and regulation with adrenal functions. PMID:27499746

  9. A novel aldose-aldose oxidoreductase for co-production of D-xylonate and xylitol from D-xylose with Saccharomyces cerevisiae.

    PubMed

    Wiebe, Marilyn G; Nygård, Yvonne; Oja, Merja; Andberg, Martina; Ruohonen, Laura; Koivula, Anu; Penttilä, Merja; Toivari, Mervi

    2015-11-01

    An open reading frame CC1225 from the Caulobacter crescentus CB15 genome sequence belongs to the Gfo/Idh/MocA protein family and has 47 % amino acid sequence identity with the glucose-fructose oxidoreductase from Zymomonas mobilis (Zm GFOR). We expressed the ORF CC1225 in the yeast Saccharomyces cerevisiae and used a yeast strain expressing the gene coding for Zm GFOR as a reference. Cell extracts of strains overexpressing CC1225 (renamed as Cc aaor) showed some Zm GFOR type of activity, producing D-gluconate and D-sorbitol when a mixture of D-glucose and D-fructose was used as substrate. However, the activity in Cc aaor expressing strain was >100-fold lower compared to strains expressing Zm gfor. Interestingly, C. crescentus AAOR was clearly more efficient than the Zm GFOR in converting in vitro a single sugar substrate D-xylose (10 mM) to xylitol without an added cofactor, whereas this type of activity was very low with Zm GFOR. Furthermore, when cultured in the presence of D-xylose, the S. cerevisiae strain expressing Cc aaor produced nearly equal concentrations of D-xylonate and xylitol (12.5 g D-xylonate l(-1) and 11.5 g D-xylitol l(-1) from 26 g D-xylose l(-1)), whereas the control strain and strain expressing Zm gfor produced only D-xylitol (5 g l(-1)). Deletion of the gene encoding the major aldose reductase, Gre3p, did not affect xylitol production in the strain expressing Cc aaor, but decreased xylitol production in the strain expressing Zm gfor. In addition, expression of Cc aaor together with the D-xylonolactone lactonase encoding the gene xylC from C. crescentus slightly increased the final concentration and initial volumetric production rate of both D-xylonate and D-xylitol. These results suggest that C. crescentus AAOR is a novel type of oxidoreductase able to convert the single aldose substrate D-xylose to both its oxidized and reduced product.

  10. Comparative anatomy of the aldo-keto reductase superfamily.

    PubMed Central

    Jez, J M; Bennett, M J; Schlegel, B P; Lewis, M; Penning, T M

    1997-01-01

    The aldo-keto reductases metabolize a wide range of substrates and are potential drug targets. This protein superfamily includes aldose reductases, aldehyde reductases, hydroxysteroid dehydrogenases and dihydrodiol dehydrogenases. By combining multiple sequence alignments with known three-dimensional structures and the results of site-directed mutagenesis studies, we have developed a structure/function analysis of this superfamily. Our studies suggest that the (alpha/beta)8-barrel fold provides a common scaffold for an NAD(P)(H)-dependent catalytic activity, with substrate specificity determined by variation of loops on the C-terminal side of the barrel. All the aldo-keto reductases are dependent on nicotinamide cofactors for catalysis and retain a similar cofactor binding site, even among proteins with less than 30% amino acid sequence identity. Likewise, the aldo-keto reductase active site is highly conserved. However, our alignments indicate that variation ofa single residue in the active site may alter the reaction mechanism from carbonyl oxidoreduction to carbon-carbon double-bond reduction, as in the 3-oxo-5beta-steroid 4-dehydrogenases (Delta4-3-ketosteroid 5beta-reductases) of the superfamily. Comparison of the proposed substrate binding pocket suggests residues 54 and 118, near the active site, as possible discriminators between sugar and steroid substrates. In addition, sequence alignment and subsequent homology modelling of mouse liver 17beta-hydroxysteroid dehydrogenase and rat ovary 20alpha-hydroxysteroid dehydrogenase indicate that three loops on the C-terminal side of the barrel play potential roles in determining the positional and stereo-specificity of the hydroxysteroid dehydrogenases. Finally, we propose that the aldo-keto reductase superfamily may represent an example of divergent evolution from an ancestral multifunctional oxidoreductase and an example of convergent evolution to the same active-site constellation as the short

  11. Some physical and immunological properties of ox kidney biliverdin reductase.

    PubMed Central

    Rigney, E M; Phillips, O; Mantle, T J

    1988-01-01

    The liver, kidney and spleen of the mouse and rat and the kidney and spleen of the ox express a monomeric form of biliverdin reductase (Mr 34,000), which in the case of the ox kidney enzyme exists in two forms (pI 5.4 and 5.2) that are probably charge isomers. The livers of the mouse and rats express, in addition, a protein (Mr 46,000) that cross-reacts with antibodies raised against the ox kidney enzyme and may be related to form 2 described by Frydman, Tomaro, Awruch & Frydman [(1983) Biochim. Biophys. Acta 759, 257-263]. Higher-Mr forms appear to exist in the guinea pig and hamster. The ox kidney enzyme has three thiol groups, of which two are accessible to 5,5'-dithiobis-(2-nitrobenzoate) in the native enzyme. Immunocytochemical analysis reveals that biliverdin reductase is localized in proximal tubules of the inner cortex of the rat kidney. Biliverdin reductase antiserum also stains proximal tubules in human and ox kidney. The staining of podocytes in glomeruli of ox kidney with antiserum to aldose reductase is particularly prominent. The localization of biliverdin reductase in the inner cortical zone of rat kidney is similar to that described for glutathione S-transferase YfYf, and it is suggested that one function of this 'intracellular binding protein' may be to maintain a low free concentration of biliverdin to allow biliverdin reductase to operate efficiently. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. PMID:3060109

  12. The C-terminal loop of aldehyde reductase determines the substrate and inhibitor specificity.

    PubMed

    Barski, O A; Gabbay, K H; Bohren, K M

    1996-11-12

    Human aldehyde reductase has a preference for carboxyl group-containing negatively charged substrates. It belongs to the NADPH-dependent aldo-keto reductase superfamily whose members are in part distinguished by unique C-terminal loops. To probe the role of the C-terminal loops in determining substrate specificities in these enzymes, two arginine residues, Arg308 and Arg311, located in the C-terminal loop of aldehyde reductase, and not found in any other C-terminal loop, were replaced with alanine residues. The catalytic efficiency of the R311A mutant for aldehydes containing a carboxyl group is reduced 150-250-fold in comparison to that of the wild-type enzyme, while substrates not containing a negative charge are unaffected. The R311A mutant is also significantly less sensitive to inhibition by dicarboxylic acids, indicating that Arg311 interacts with one of the carboxyl groups. The inhibition pattern indicates that the other carboxyl group binds to the anion binding site formed by Tyr49, His112, and the nicotinamide moiety of NADP+. The correlation between inhibitor potency and the length of the dicarboxylic acid molecules suggests a distance of approximately 10 A between the amino group of Arg311 and the anion binding site in the aldehyde reductase molecule. The sensitivity of inhibition of the R311A mutant by several commercially available aldose reductase inhibitors (ARIs) was variable, with tolrestat and zopolrestat becoming more potent inhibitors (30- and 5-fold, respectively), while others remained the same or became less potent. The catalytic properties, substrate specificity, and susceptibility to inhibition of the R308A mutant remained similar to that of the wild-type enzyme. The data provide direct evidence for C-terminal loop participation in determining substrate and inhibitor specificity of aldo-keto reductases and specifically identifies Arg311 as the basis for the carboxyl-containing substrate preference of aldehyde reductase. PMID:8916913

  13. arsRBOCT Arsenic Resistance System Encoded by Linear Plasmid pHZ227 in Streptomyces sp. Strain FR-008

    PubMed Central

    Wang, Lianrong; Chen, Shi; Xiao, Xiang; Huang, Xi; You, Delin; Zhou, Xiufen; Deng, Zixin

    2006-01-01

    In the arsenic resistance gene cluster from the large linear plasmid pHZ227, two novel genes, arsO (for a putative flavin-binding monooxygenase) and arsT (for a putative thioredoxin reductase), were coactivated and cotranscribed with arsR1-arsB and arsC, respectively. Deletion of the ars gene cluster on pHZ227 in Streptomyces sp. strain FR-008 resulted in sensitivity to arsenic, and heterologous expression of the ars gene cluster in the arsenic-sensitive Streptomyces strains conferred resistance on the new hosts. The pHZ227 ArsB protein showed homology to the yeast arsenite transporter Acr3p. The pHZ227 ArsC appears to be a bacterial thioredoxin-dependent ArsC-type arsenate reductase with four conserved cysteine thioredoxin-requiring motifs. PMID:16672525

  14. Quinone Reductase 2 Is a Catechol Quinone Reductase

    SciTech Connect

    Fu, Yue; Buryanovskyy, Leonid; Zhang, Zhongtao

    2008-09-05

    The functions of quinone reductase 2 have eluded researchers for decades even though a genetic polymorphism is associated with various neurological disorders. Employing enzymatic studies using adrenochrome as a substrate, we show that quinone reductase 2 is specific for the reduction of adrenochrome, whereas quinone reductase 1 shows no activity. We also solved the crystal structure of quinone reductase 2 in complexes with dopamine and adrenochrome, two compounds that are structurally related to catecholamine quinones. Detailed structural analyses delineate the mechanism of quinone reductase 2 specificity toward catechol quinones in comparison with quinone reductase 1; a side-chain rotational difference between quinone reductase 1 and quinone reductase 2 of a single residue, phenylalanine 106, determines the specificity of enzymatic activities. These results infer functional differences between two homologous enzymes and indicate that quinone reductase 2 could play important roles in the regulation of catecholamine oxidation processes that may be involved in the etiology of Parkinson disease.

  15. Base catalysed isomerisation of aldoses of the arabino and lyxo series in the presence of aluminate.

    PubMed

    Ekeberg, Dag; Morgenlie, Svein; Stenstrøm, Yngve

    2002-04-30

    Base-catalysed isomerisation of aldoses of the arabino and lyxo series in aluminate solution has been investigated. L-Arabinose and D-galactose give L-erythro-2-pentulose (L-ribulose) and D-lyxo-2-hexulose (D-tagatose), respectively, in good yields, whereas lower reactivity is observed for 6-deoxy-D-galactose (D-fucose). From D-lyxose, D-mannose and 6-deoxy-L-mannose (L-rhamnose) are obtained mixtures of ketoses and C-2 epimeric aldoses. Small amounts of the 3-epimers of the ketoses were also formed. 6-Deoxy-L-arabino-2-hexulose (6-deoxy-L-fructose) and 6-deoxy-L-glucose (L-quinovose) were formed in low yields from 6-deoxy-L-mannose and isolated as their O-isopropylidene derivatives. Explanations of the differences in reactivity and course of the reaction have been suggested on the basis of steric effects.

  16. Ar-Ar_Redux: rigorous error propagation of 40Ar/39Ar data, including covariances

    NASA Astrophysics Data System (ADS)

    Vermeesch, P.

    2015-12-01

    Rigorous data reduction and error propagation algorithms are needed to realise Earthtime's objective to improve the interlaboratory accuracy of 40Ar/39Ar dating to better than 1% and thereby facilitate the comparison and combination of the K-Ar and U-Pb chronometers. Ar-Ar_Redux is a new data reduction protocol and software program for 40Ar/39Ar geochronology which takes into account two previously underappreciated aspects of the method: 1. 40Ar/39Ar measurements are compositional dataIn its simplest form, the 40Ar/39Ar age equation can be written as: t = log(1+J [40Ar/39Ar-298.5636Ar/39Ar])/λ = log(1 + JR)/λ Where λ is the 40K decay constant and J is the irradiation parameter. The age t does not depend on the absolute abundances of the three argon isotopes but only on their relative ratios. Thus, the 36Ar, 39Ar and 40Ar abundances can be normalised to unity and plotted on a ternary diagram or 'simplex'. Argon isotopic data are therefore subject to the peculiar mathematics of 'compositional data', sensu Aitchison (1986, The Statistical Analysis of Compositional Data, Chapman & Hall). 2. Correlated errors are pervasive throughout the 40Ar/39Ar methodCurrent data reduction protocols for 40Ar/39Ar geochronology propagate the age uncertainty as follows: σ2(t) = [J2 σ2(R) + R2 σ2(J)] / [λ2 (1 + R J)], which implies zero covariance between R and J. In reality, however, significant error correlations are found in every step of the 40Ar/39Ar data acquisition and processing, in both single and multi collector instruments, during blank, interference and decay corrections, age calculation etc. Ar-Ar_Redux revisits every aspect of the 40Ar/39Ar method by casting the raw mass spectrometer data into a contingency table of logratios, which automatically keeps track of all covariances in a compositional context. Application of the method to real data reveals strong correlations (r2 of up to 0.9) between age measurements within a single irradiation batch. Propertly taking

  17. Prokaryotic arsenate reductase enhances arsenate resistance in Mammalian cells.

    PubMed

    Wu, Dan; Tao, Xuanyu; Wu, Gaofeng; Li, Xiangkai; Liu, Pu

    2014-01-01

    Arsenic is a well-known heavy metal toxicant in the environment. Bioremediation of heavy metals has been proposed as a low-cost and eco-friendly method. This article described some of recent patents on transgenic plants with enhanced heavy metal resistance. Further, to test whether genetic modification of mammalian cells could render higher arsenic resistance, a prokaryotic arsenic reductase gene arsC was transfected into human liver cancer cell HepG2. In the stably transfected cells, the expression level of arsC gene was determined by quantitative real-time PCR. Results showed that arsC was expressed in HepG2 cells and the expression was upregulated by 3 folds upon arsenate induction. To further test whether arsC has function in HepG2 cells, the viability of HepG2-pCI-ArsC cells exposed to arsenite or arsenate was compared to that of HepG2-pCI cells without arsC gene. The results indicated that arsC increased the viability of HepG2 cells by 25% in arsenate, but not in arsenite. And the test of reducing ability of stably transfected cells revealed that the concentration of accumulated trivalent arsenic increased by 25% in HepG2-pCI-ArsC cells. To determine the intracellular localization of ArsC, a fusion vector with fluorescent marker pEGFP-N1-ArsC was constructed and transfected into.HepG2. Laser confocal microscopy showed that EGFP-ArsC fusion protein was distributed throughout the cells. Taken together, these results demonstrated that prokaryotic arsenic resistant gene arsC integrated successfully into HepG2 genome and enhanced arsenate resistance of HepG2, which brought new insights of arsenic detoxification in mammalian cells.

  18. Differential expression of 5-alpha reductase isozymes in the prostate and its clinical implications

    PubMed Central

    Wang, Kai; Fan, Dong-Dong; Jin, Song; Xing, Nian-Zeng; Niu, Yi-Nong

    2014-01-01

    The development of human benign or malignant prostatic diseases is closely associated with androgens, primarily testosterone (T) and dihydrotestosterone (DHT). T is converted to DHT by 5-alpha reductase (5-AR) isozymes. Differential expression of 5-AR isozymes is observed in both human benign and malignant prostatic tissues. 5-AR inhibitors (5-ARI) are commonly used for the treatment of benign prostatic hyperplasia (BPH) and were once promoted as chemopreventive agents for prostate cancer (PCa). This review discusses the role of the differential expression of 5-AR in the normal development of the human prostate and in the pathogenesis and progression of BPH and PCa. PMID:24457841

  19. An ethoxyquin-inducible aldehyde reductase from rat liver that metabolizes aflatoxin B1 defines a subfamily of aldo-keto reductases.

    PubMed

    Ellis, E M; Judah, D J; Neal, G E; Hayes, J D

    1993-11-01

    Protection of liver against the toxic and carcinogenic effects of aflatoxin B1 (AFB1) can be achieved through the induction of detoxification enzymes by chemoprotectors such as the phenolic antioxidant ethoxyquin. We have cloned and sequenced a cDNA encoding an aldehyde reductase (AFB1-AR), which is expressed in rat liver in response to dietary ethoxyquin. Expression of the cDNA in Escherichia coli and purification of the recombinant enzyme reveals that the protein exhibits aldehyde reductase activity and is capable of converting the protein-binding dialdehyde form of AFB1-dihydrodiol to the nonbinding dialcohol metabolite. We show that the mRNA encoding this enzyme is markedly elevated in the liver of rats fed an ethoxyquin-containing diet, correlating with acquisition of resistance to AFB1. AFB1-AR represents the only carcinogen-metabolizing aldehyde reductase identified to date that is induced by a chemoprotector. Alignment of the amino acid sequence of AFB1-AR with other known and putative aldehyde reductases shows that it defines a subfamily within the aldo-keto reductase superfamily. PMID:8234296

  20. A Novel Aldo-Keto Reductase, HdRed, from the Pacific Abalone Haliotis discus hannai, Which Reduces Alginate-derived 4-Deoxy-L-erythro-5-hexoseulose Uronic Acid to 2-Keto-3-deoxy-D-gluconate.

    PubMed

    Mochizuki, Shogo; Nishiyama, Ryuji; Inoue, Akira; Ojima, Takao

    2015-12-25

    Abalone feeds on brown seaweeds and digests seaweeds' alginate with alginate lyases (EC 4.2.2.3). However, it has been unclear whether the end product of alginate lyases (i.e. unsaturated monouronate-derived 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH)) is assimilated by abalone itself, because DEH cannot be metabolized via the Embden-Meyerhof pathway of animals. Under these circumstances, we recently noticed the occurrence of an NADPH-dependent reductase, which reduced DEH to 2-keto-3-deoxy-D-gluconate, in hepatopancreas extract of the pacific abalone Haliotis discus hannai. In the present study, we characterized this enzyme to some extent. The DEH reductase, named HdRed in the present study, could be purified from the acetone-dried powder of hepatopancreas by ammonium sulfate fractionation followed by conventional column chromatographies. HdRed showed a single band of ∼ 40 kDa on SDS-PAGE and reduced DEH to 2-keto-3-deoxy-D-gluconate with an optimal temperature and pH at around 50 °C and 7.0, respectively. HdRed exhibited no appreciable activity toward 28 authentic compounds, including aldehyde, aldose, ketose, α-keto-acid, uronic acid, deoxy sugar, sugar alcohol, carboxylic acid, ketone, and ester. The amino acid sequence of 371 residues of HdRed deduced from the cDNA showed 18-60% identities to those of aldo-keto reductase (AKR) superfamily enzymes, such as human aldose reductase, halophilic bacterium reductase, and sea hare norsolorinic acid (a polyketide derivative) reductase-like protein. Catalytic residues and cofactor binding residues known in AKR superfamily enzymes were fairly well conserved in HdRed. Phylogenetic analysis for HdRed and AKR superfamily enzymes indicated that HdRed is an AKR belonging to a novel family. PMID:26555267

  1. A Novel Aldo-Keto Reductase, HdRed, from the Pacific Abalone Haliotis discus hannai, Which Reduces Alginate-derived 4-Deoxy-L-erythro-5-hexoseulose Uronic Acid to 2-Keto-3-deoxy-D-gluconate.

    PubMed

    Mochizuki, Shogo; Nishiyama, Ryuji; Inoue, Akira; Ojima, Takao

    2015-12-25

    Abalone feeds on brown seaweeds and digests seaweeds' alginate with alginate lyases (EC 4.2.2.3). However, it has been unclear whether the end product of alginate lyases (i.e. unsaturated monouronate-derived 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH)) is assimilated by abalone itself, because DEH cannot be metabolized via the Embden-Meyerhof pathway of animals. Under these circumstances, we recently noticed the occurrence of an NADPH-dependent reductase, which reduced DEH to 2-keto-3-deoxy-D-gluconate, in hepatopancreas extract of the pacific abalone Haliotis discus hannai. In the present study, we characterized this enzyme to some extent. The DEH reductase, named HdRed in the present study, could be purified from the acetone-dried powder of hepatopancreas by ammonium sulfate fractionation followed by conventional column chromatographies. HdRed showed a single band of ∼ 40 kDa on SDS-PAGE and reduced DEH to 2-keto-3-deoxy-D-gluconate with an optimal temperature and pH at around 50 °C and 7.0, respectively. HdRed exhibited no appreciable activity toward 28 authentic compounds, including aldehyde, aldose, ketose, α-keto-acid, uronic acid, deoxy sugar, sugar alcohol, carboxylic acid, ketone, and ester. The amino acid sequence of 371 residues of HdRed deduced from the cDNA showed 18-60% identities to those of aldo-keto reductase (AKR) superfamily enzymes, such as human aldose reductase, halophilic bacterium reductase, and sea hare norsolorinic acid (a polyketide derivative) reductase-like protein. Catalytic residues and cofactor binding residues known in AKR superfamily enzymes were fairly well conserved in HdRed. Phylogenetic analysis for HdRed and AKR superfamily enzymes indicated that HdRed is an AKR belonging to a novel family.

  2. Sequence analysis of frog rho-crystallin by cDNA cloning and sequencing: a member of the aldo-keto reductase family.

    PubMed

    Lu, S F; Pan, F M; Chiou, S H

    1995-09-25

    rho-Crystallin is a major enzyme crystallin present in the lenses of amphibian species with a blocked amino terminus. In order to facilitate the determination of the primary sequence of this taxon-specific crystallin, cDNA mixture was synthesized from the poly(A)+mRNA of bullfrog eye lenses. cDNAs encoding rho-crystallin were then amplified by polymerase chain reaction (PCR) using a new protocol of Rapid Amplification of cDNA Ends (RACE). PCR-amplified product corresponding to rho-crystallin was obtained, which was then subcloned into pUC18 vector and then transformed into E. coli strain JM109. Plasmids purified from the positive clones were prepared for nucleotide sequencing by the automatic fluorescence-based dideoxynucleotide chain-termination method. Sequencing more than 15 clones containing DNA inserts coding for rho-crystallin constructed only one unique and complete full-length reading frame of 975 base pairs covering a deduced protein sequence of 324 amino acids including the universal initiating methionine. It shows 96, 59, 46 and 37 percent sequence similarity to another rho-crystallin from European common frog, bovine prostaglandin-F synthase, human aldose reductase and human aldehyde reductase, respectively, revealing the close relationship between rho-crystallins from related amphibian species and its possible evolutionary relatedness with various aldo-keto reductases. In this study a phylogenetic tree for rho-crystallin and related enzymes is constructed based on multiple-sequence alignment program using a combination of distance matrix and approximate parsimony methods. We have thus established the remote phylogenetic relationship between rho-crystallin and some aldehyde/aldose reductases, which may provide a possible link for the recruitment of this crystallin from detoxification-related enzymes and its physiological role in maintaining a transparent and clear lens.

  3. An Expeditious Synthesis of Sialic Acid Derivatives by Copper(I)-Catalyzed Stereodivergent Propargylation of Unprotected Aldoses

    PubMed Central

    2016-01-01

    We developed a copper(I)-catalyzed stereodivergent anomeric propargylation of unprotected aldoses as a facile synthetic pathway to a broad variety of sialic acid derivatives. The soft allenylcopper(I) species, catalytically generated from stable allenylboronic acid pinacolate (2), is unusually inert to protonolysis by the multiple hydroxy groups of the substrates and thereby functions as a carbon nucleophile. The key additive B(OMe)3 facilitated ring-opening of the nonelectrophilic cyclic hemiacetal forms of aldoses to the reactive aldehyde forms. The chirality of the catalyst, and not the internal stereogenic centers of substrates, predominantly controlled the stereochemistry of the propargylation step; i.e., the diastereoselectivity was switched simply by changing the catalyst chirality. This is the first nonenzyme catalyst-controlled stereodivergent C–C bond elongation at the anomeric center of unprotected aldoses, which contain multiple protic functional groups and stereogenic centers. The propargylation products can be expeditiously transformed into naturally occurring and synthetic sialic acid derivatives in a simple three-step sequence. This synthetic method, which requires no protecting groups, can be performed on a gram-scale and thus offers general and practical access to various sialic acid derivatives from unprotected aldoses. PMID:27163022

  4. An Expeditious Synthesis of Sialic Acid Derivatives by Copper(I)-Catalyzed Stereodivergent Propargylation of Unprotected Aldoses.

    PubMed

    Wei, Xiao-Feng; Shimizu, Yohei; Kanai, Motomu

    2016-01-27

    We developed a copper(I)-catalyzed stereodivergent anomeric propargylation of unprotected aldoses as a facile synthetic pathway to a broad variety of sialic acid derivatives. The soft allenylcopper(I) species, catalytically generated from stable allenylboronic acid pinacolate (2), is unusually inert to protonolysis by the multiple hydroxy groups of the substrates and thereby functions as a carbon nucleophile. The key additive B(OMe)3 facilitated ring-opening of the nonelectrophilic cyclic hemiacetal forms of aldoses to the reactive aldehyde forms. The chirality of the catalyst, and not the internal stereogenic centers of substrates, predominantly controlled the stereochemistry of the propargylation step; i.e., the diastereoselectivity was switched simply by changing the catalyst chirality. This is the first nonenzyme catalyst-controlled stereodivergent C-C bond elongation at the anomeric center of unprotected aldoses, which contain multiple protic functional groups and stereogenic centers. The propargylation products can be expeditiously transformed into naturally occurring and synthetic sialic acid derivatives in a simple three-step sequence. This synthetic method, which requires no protecting groups, can be performed on a gram-scale and thus offers general and practical access to various sialic acid derivatives from unprotected aldoses. PMID:27163022

  5. Nitrate and periplasmic nitrate reductases

    PubMed Central

    Sparacino-Watkins, Courtney; Stolz, John F.; Basu, Partha

    2014-01-01

    The nitrate anion is a simple, abundant and relatively stable species, yet plays a significant role in global cycling of nitrogen, global climate change, and human health. Although it has been known for quite some time that nitrate is an important species environmentally, recent studies have identified potential medical applications. In this respect the nitrate anion remains an enigmatic species that promises to offer exciting science in years to come. Many bacteria readily reduce nitrate to nitrite via nitrate reductases. Classified into three distinct types – periplasmic nitrate reductase (Nap), respiratory nitrate reductase (Nar) and assimilatory nitrate reductase (Nas), they are defined by their cellular location, operon organization and active site structure. Of these, Nap proteins are the focus of this review. Despite similarities in the catalytic and spectroscopic properties Nap from different Proteobacteria are phylogenetically distinct. This review has two major sections: in the first section, nitrate in the nitrogen cycle and human health, taxonomy of nitrate reductases, assimilatory and dissimilatory nitrate reduction, cellular locations of nitrate reductases, structural and redox chemistry are discussed. The second section focuses on the features of periplasmic nitrate reductase where the catalytic subunit of the Nap and its kinetic properties, auxiliary Nap proteins, operon structure and phylogenetic relationships are discussed. PMID:24141308

  6. Investigation of the functional role of aldose 1-epimerase in engineered cellobiose utilization.

    PubMed

    Li, Sijin; Ha, Suk-Jin; Kim, Hee Jin; Galazka, Jonathan M; Cate, Jamie H D; Jin, Yong-Su; Zhao, Huimin

    2013-10-10

    Functional expression of a cellodextrin transporter and an intracellular β-glucosidase from Neurospora crassa in Saccharomyces cerevisiae enables simultaneous co-fermentation of cellobiose and non-glucose sugars such as xylose. Here we investigate the functional role of aldose 1-epimerase (AEP) in engineered cellobiose utilization. One AEP (Gal10) and two putative AEPs (Yhr210c and Ynr071c sharing 50.6% and 51.0% amino acid identity with Gal10, respectively) were selected. Deletion of GAL10 led to complete loss of both AEP activity and cell growth on cellobiose, while GAL10 complementation restored the AEP activity and cell growth. In addition, deletion of YHR210C or YNR071C resulted in improved cellobiose utilization. These results suggest that the intracellular mutarotation of β-glucose to α-glucose might be a rate controlling step and Gal10 play a crucial role in cellobiose fermentation by engineered S. cerevisiae. PMID:23954547

  7. Asymmetric assembly of aldose carbohydrates from formaldehyde and glycolaldehyde by tandem biocatalytic aldol reactions

    NASA Astrophysics Data System (ADS)

    Szekrenyi, Anna; Garrabou, Xavier; Parella, Teodor; Joglar, Jesús; Bujons, Jordi; Clapés, Pere

    2015-09-01

    The preparation of multifunctional chiral molecules can be greatly simplified by adopting a route via the sequential catalytic assembly of achiral building blocks. The catalytic aldol assembly of prebiotic compounds into stereodefined pentoses and hexoses is an as yet unmet challenge. Such a process would be of remarkable synthetic utility and highly significant with regard to the origin of life. Pursuing an expedient enzymatic approach, here we use engineered D-fructose-6-phosphate aldolase from Escherichia coli to prepare a series of three- to six-carbon aldoses by sequential one-pot additions of glycolaldehyde. Notably, the pertinent selection of the aldolase variant provides control of the sugar size. The stereochemical outcome of the addition was also altered to allow the synthesis of L-glucose and related derivatives. Such engineered biocatalysts may offer new routes for the straightforward synthesis of natural molecules and their analogues that circumvent the intricate enzymatic pathways forged by evolution.

  8. Human and rodent aldo-keto reductases from the AKR1B subfamily and their specificity with retinaldehyde

    PubMed Central

    Ruiz, F. Xavier; Moro, Armando; Gallego, Oriol; Ardèvol, Albert; Rovira, Carme; Petrash, J. Mark; Parés, Xavier; Farrés, Jaume

    2011-01-01

    NADP(H)-dependent cytosolic aldo-keto reductases (AKR) are mostly monomeric enzymes which fold into a typical (α/β)8-barrel structure. Substrate specificity and inhibitor selectivity are determined by interaction with residues located in three highly variable loops (A, B, and C). Based on sequence identity, AKR have been grouped into families, namely AKR1–AKR15, containing multiple subfamilies. Two human enzymes from the AKR1B subfamily (AKR1B1 and AKR1B10) are of special interest. AKR1B1 (aldose reductase) is related to secondary diabetic complications, while AKR1B10 is induced in cancer cells and is highly active with all-trans-retinaldehyde. Residues interacting with all-trans-retinaldehyde and differing between AKR1B1 and AKR1B10 are Leu125Lys and Val131Ala (loop A), Leu301Val, Ser303Gln, and Cys304Ser (loop C). Recently, we demonstrated the importance of Lys125 as a determinant of AKR1B10 specificity for retinoids. Residues 301 and 304 are also involved in interactions with substrates or inhibitors, and thus we checked their contribution to retinoid specificity. We also extended our study with retinoids to rodent members of the AKR1B subfamily: AKR1B3 (aldose reductase), AKR1B7 (mouse vas deferens protein), AKR1B8 (fibroblast-growth factor 1-regulated protein), and AKR1B9 (Chinese hamster ovary reductase), which were tested against all-trans isomers of retinaldehyde and retinol. All enzymes were active with retinaldehyde, but with kcat values (0.02–0.52 min−1) much lower than that of AKR1B10 (27 min−1). None of the enzymes showed oxidizing activity with retinol. Since these enzymes (except AKR1B3) have Lys125, other residues should account for retinaldehyde specificity. Here, by using site-directed mutagenesis and molecular modeling, we further delineate the contribution of residues 301 and 304. We demonstrate that besides Lys125, Ser304 is a major structural determinant for all-trans-retinaldehyde specificity of AKR1B10. PMID:21329680

  9. Isolated menthone reductase and nucleic acid molecules encoding same

    DOEpatents

    Croteau, Rodney B; Davis, Edward M; Ringer, Kerry L

    2013-04-23

    The present invention provides isolated menthone reductase proteins, isolated nucleic acid molecules encoding menthone reductase proteins, methods for expressing and isolating menthone reductase proteins, and transgenic plants expressing elevated levels of menthone reductase protein.

  10. Molecular distribution and degradation status of combined aldoses in sinking particulate organic matter

    NASA Astrophysics Data System (ADS)

    Panagiotopoulos, C.; Sempéré, R.

    2003-04-01

    Particulate samples were collected by using floating sediment traps (50--300 m) and in situ pumps (30 and 200 m) in the Southern Indian Ocean (Polar Front Zone (PFZ) and Sub-Tropical Zone (STZ)), Mediterranean Sea (Ligurian and Ionian Seas) and Atlantic Ocean (Upwelling (UPW) of Agadir-Morocco). They were studied for monosaccharide composition after acid hydrolysis (HCl 0.09 M, 20 h, 100^oC) by using High Performance Anion Exchange Chromatography followed by Pulsed Amperometric Detection (HPAEC-PAD). Our results indicated that higher PCHO yields (calculated as PCHO-C/POC ratios) were associated to higher C:N ratios (Med. Sea sample, PCHO yields = 12.7 ± 7.7%; C:N ratios = 8.3 ± 1.6; n = 12) whether the opposite trend was found for Southern Ocean samples (PCHO yields = 3.3 ± 0.75%; C:N ratios = 5.7 ± 0.59, n = 5) indicating significant variability in the sugar content of particles which might be due to the degradation degree of the particles as well as to the initial chemical composition of plankton. Alternatively, other processes such as high production of extracellular polysaccharides (type transparent exopolymer polysaccharides (TEP)) due to phosphorus limitation of some phytoplanktonic species may increase the sugar content in Mediterranean particles and the C/N ratio. In any case, glucose appeared to be the most abundant monosaccharide in Mediterranean Sea or UPW samples (range 23--59 wt% of the total aldoses) whereas ribose (17--39 wt%) and galactose (range 10--28 wt%) were the predominant aldoses in Southern Indian Ocean. These sugars (glucose + ribose) exhibited a strong negative relationship with C:N (r = -0.53, p >0.01; n = 30) in sediment traps (data from this study) and sediment (data from literature) particulate material which further indicates that these two monosaccharides are selectively extracted from the carbohydrate pool in sediment. In vitro biodegradation experiments performed with large particles (>60 μm) sampled using in situ pumps in

  11. Zeatin reductase in Phaseolus embryos

    SciTech Connect

    Martin, R.C.; Mok, David, W.S.; Mok, M.C. )

    1989-04-01

    Zeatin was converted to O-xylosylzeatin in embryos of Phaseolus vulgaris . O-xylosyldihydrozeatin was also identified as a zeatin metabolite. Incubation of embryo extracts with {sup 14}C-zeatin and {sup 14}C-O-xylosylzeatin revealed that reduction preceeds the O-xylosylation of zeatin. An enzyme responsible for reducing the N{sup 6}-side chain was isolated and partially purified using ammonium sulfate fractionation and affinity, gel filtration and anion exchange chromatography. The NADPH dependent reductase was zeatin specific and did not recognize cis-zeatin, ribosylzeatin, i{sup 6}Ade or i{sup 6}Ado. Two forms of the reductase could be separated by either gel filtration or anion exchange HPLC. The HMW isozyme (Mr. 55,000) eluted from the anion exchange column later than the LMW isozyme (Mr. 25,000). Interspecific differences in zeatin reductase activity were also detected.

  12. Genetics Home Reference: 5-alpha reductase deficiency

    MedlinePlus

    ... gene provides instructions for making an enzyme called steroid 5-alpha reductase 2. This enzyme is involved ... external genitalia. Mutations in the SRD5A2 gene prevent steroid 5-alpha reductase 2 from effectively converting testosterone ...

  13. Ar-Ar ages and thermal histories of enstatite meteorites

    NASA Astrophysics Data System (ADS)

    Bogard, Donald D.; Dixon, Eleanor T.; Garrison, Daniel H.

    2010-05-01

    Compared with ordinary chondrites, there is a relative paucity of chronological and other data to define the early thermal histories of enstatite parent bodies. In this study, we report 39Ar-40Ar dating results for five EL chondrites: Khairpur, Pillistfer, Hvittis, Blithfield, and Forrest; five EH chondrites: Parsa, Saint Marks, Indarch, Bethune, and Reckling Peak 80259; three igneous-textured enstatite meteorites that represent impact melts on enstatite chondrite parent bodies: Zaklodzie, Queen Alexandra Range 97348, and Queen Alexandra Range 97289; and three aubrites, Norton County, Bishopville, and Cumberland Falls Several Ar-Ar age spectra show unusual 39Ar recoil effects, possibly the result of some of the K residing in unusual sulfide minerals, such as djerfisherite and rodderite, and other age spectra show 40Ar diffusion loss. Few additional Ar-Ar ages for enstatite meteorites are available in the literature. When all available Ar-Ar data on enstatite meteorites are considered, preferred ages of nine chondrites and one aubrite show a range of 4.50-4.54Ga, whereas five other meteorites show only lower age limits over 4.35-4.46Ga. Ar-Ar ages of several enstatite chondrites are as old or older as the oldest Ar-Ar ages of ordinary chondrites, which suggests that enstatite chondrites may have derived from somewhat smaller parent bodies, or were metamorphosed to lower temperatures compared to other chondrite types. Many enstatite meteorites are brecciated and/or shocked, and some of the younger Ar-Ar ages may record these impact events. Although impact heating of ordinary chondrites within the last 1Ga is relatively common for ordinary chondrites, only Bethune gives any significant evidence for such a young event.

  14. The binding sites on human heme oxygenase-1 for cytochrome p450 reductase and biliverdin reductase.

    PubMed

    Wang, Jinling; de Montellano, Paul R Ortiz

    2003-05-30

    Human heme oxygenase-1 (hHO-1) catalyzes the NADPH-cytochrome P450 reductase-dependent oxidation of heme to biliverdin, CO, and free iron. The biliverdin is subsequently reduced to bilirubin by biliverdin reductase. Earlier kinetic studies suggested that biliverdin reductase facilitates the release of biliverdin from hHO-1 (Liu, Y., and Ortiz de Montellano, P. R. (2000) J. Biol. Chem. 275, 5297-5307). We have investigated the binding of P450 reductase and biliverdin reductase to truncated, soluble hHO-1 by fluorescence resonance energy transfer and site-specific mutagenesis. P450 reductase and biliverdin reductase bind to truncated hHO-1 with Kd = 0.4 +/- 0.1 and 0.2 +/- 0.1 microm, respectively. FRET experiments indicate that biliverdin reductase and P450 reductase compete for binding to truncated hHO-1. Mutation of surface ionic residues shows that hHO-1 residues Lys18, Lys22, Lys179, Arg183, Arg198, Glu19, Glu127, and Glu190 contribute to the binding of cytochrome P450 reductase. The mutagenesis results and a computational analysis of the protein surfaces partially define the binding site for P450 reductase. An overlapping binding site including Lys18, Lys22, Lys179, Arg183, and Arg185 is similarly defined for biliverdin reductase. These results confirm the binding of biliverdin reductase to hHO-1 and define binding sites of the two reductases.

  15. Ar/Ar Dating Independent of Monitor Standard Ages

    NASA Astrophysics Data System (ADS)

    Boswell, S.; Hemming, S. R.

    2015-12-01

    Because the reported age of an analyzed sample is dependent on the age of the co-irradiated monitor standard(s), Ar/Ar dating is a relative dating technique. There is disagreement at the 1% scale in the age of commonly used monitor standards, and there is a great need to improve the inter-laboratory calibrations. Additionally, new approaches and insights are needed to meet the challenge of bringing the Ar/Ar chronometer to the highest possible precision and accuracy. In this spirit, we present a conceptual framework for Ar/Ar dating that does not depend on the age of monitor standards, but only on the K content of a solid standard. The concept is demonstrated by introducing a re-expressed irradiation parameter (JK) that depends on the ratio of 39ArK to 40Ar* rather than the 40Ar*/39ArK ratio. JK is equivalent to the traditional irradiation parameter J and is defined as JK = (39Ar/40K) • (λ/λe). The ultimate precision and accuracy of the method will depend on how precisely and accurately the 39Ar and 40K can be estimated, and will require isotope dilution measurements of both from the same aliquot. We are testing the workability of our technique at the 1% level by measuring weighed and irradiated hornblende and biotite monitor standards using GLO-1 glauconite to define a calibration curve for argon signals versus abundance.

  16. 40Ar/39Ar Dating of Volcanic Glass

    NASA Astrophysics Data System (ADS)

    Morgan, L. E.; Renne, P. R.; Watkins, J. M.

    2007-12-01

    Application of the 40Ar/39Ar method to volcanic glasses has been somewhat stigmatized following several studies demonstrating secondary mobility of K and Ar. Much of the stigma is unwarranted, however, since most studies only impugned the reliability of the K-Ar and 40Ar/39Ar techniques when applied to glass shards rather than obsidian clasts with low surface area to volume ratios. We provide further evidence for problematic K loss and/or 39Ar recoil ejection from glass shards in 40Ar/39Ar step heating results for comagmatic feldspars and shards. In an extreme case, the plateau age of the feldspars (0.17 ± 0.03 Ma at 2σ) is significantly younger than the plateau age of the glass (0.85 ± 0.05 Ma at 2σ). If the feldspar age is reasonably interpreted as the eruption age of the ash, it is likely that the glass shards experienced K and/or 39Ar loss. Electron microprobe analyses of the glass shards have low totals (~93%) and no systematic lateral variability (i.e., diffusion gradients) in K, suggesting that the lengthscale of the glass shards is smaller than the lengthscale of K diffusion. Obsidian clasts should not be as susceptible to K loss since any hydrated (K-depleted) volume represents a small fraction of the total material and can often be physically removed prior to analysis. Samples described here are detrital obsidian clasts from the Afar region of Ethiopia. Evidence from Fourier Transform Infrared Spectroscopy (FTIR), and previous work by Anovitz (1999), confirm that the scale of water and potassium mobility are often small in comparison to the size of obsidian clasts but large enough to effect the bulk composition of glass shards. This expectation is confirmed in another tuff wherein comagmatic obsidian clasts and sanidine phenocrysts yield indistinguishable 40Ar/39Ar ages of 4.4 Ma High abundances of non-radiogenic 40Ar, and kinetic fractionation of Ar isotopes during quenching and/or laboratory degassing resulting in incomplete equilibration between

  17. Ribonucleotide Reductase-- a Radical Enzyme

    NASA Astrophysics Data System (ADS)

    Reichard, Peter; Ehrenberg, Anders

    1983-08-01

    Ribonucleotide reductases catalyze the enzymatic formation of deoxyribonucleotides, an obligatory step in DNA synthesis. The native form of the enzyme from Escherichia coli or from mammalian sources contains as part of its polypeptide structure a free tyrosyl radical, stabilized by an iron center. The radical participates in all probability in the catalytic process during the substitution of the hydroxyl group at C-2 of ribose by a hydrogen atom. A second, inactive form of the E. coli reductase lacks the tyrosyl radical. Extracts from E. coli contain activities that interconvert the two forms. The tyrosyl radical is introduced in the presence of oxygen, while anaerobiosis favors its removal, suggesting a regulatory role in DNA synthesis for oxygen.

  18. Nitrate reductase from Rhodopseudomonas sphaeroides.

    PubMed Central

    Kerber, N L; Cardenas, J

    1982-01-01

    The facultative phototroph Rhodopseudomonas sphaeroides DSM158 was incapable of either assimilating or dissimilating nitrate, although the organism could reduce it enzymatically to nitrite either anaerobically in the light or aerobically in the dark. Reduction of nitrate was mediated by a nitrate reductase bound to chromatophores that could be easily solubilized and functioned with chemically reduced viologens or photochemically reduced flavins as electron donors. The enzyme was solubilized, and some of its kinetic and molecular parameters were determined. It seemed to be nonadaptive, ammonia did not repress its synthesis, and its activity underwent a rapid decline when the cells entered the stationary growth phase. Studies with inhibitors and with metal antagonists indicated that molybdenum and possibly iron participate in the enzymatic reduction of nitrate. The conjectural significance of this nitrate reductase in phototrophic bacteria is discussed. PMID:6978883

  19. LC-MS-MS Characterization of Forced Degradation Products of Fidarestat, a Novel Aldose Reductase Inhibitor: Development and Validation of a Stability-Indicating RP-HPLC Method.

    PubMed

    Talluri, M V N Kumar; Khatoon, Lubna; Kalariya, Pradipbhai D; Chavan, Balasaheb B; Ragampeta, Srinivas

    2015-10-01

    An accurate, precise, robust and selective stability-indicating liquid chromatographic (LC) method has been developed for the monitoring of fidarestat in the presence of its forced degradants. The drug was subjected to hydrolysis (acid, alkali and neutral degradation), oxidation, photolysis and thermal stress conditions. The drug degraded significantly under hydrolytic (basic, acidic and neutral) and oxidative stress conditions, whereas it was found to be stable in photolytic and thermal conditions. The chromatographic separation was achieved on a Grace C18, (250 mm × 4.6 mm × 5 μm) column using gradient mobile phase system consisting of 10 mM of ammonium acetate buffer at pH 4 and acetonitrile at a flow rate of 1 mL/min with UV detection at 283 nm. The developed method was extended to liquid chromatography quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS-MS) for characterization of all the degradation products. A total of five new degradation products were identified and characterized by LC-QTOF-MS-MS. The developed LC method was validated as per ICH guideline Q2 (R1). The proposed method was found to be successively applied for the quality control of fidarestat in bulk drug analysis.

  20. Targeting 5α-reductase for prostate cancer prevention and treatment

    PubMed Central

    Nacusi, Lucas P.; Tindall, Donald J.

    2014-01-01

    Testosterone is the most abundant circulating androgen, and can be converted to dihydrotestosterone (DHT), a more potent androgen, by the 5α-reductase enzymes in target tissues. Current treatments for prostate cancer consist of reducing androgen levels by chemical or surgical castration or pure antiandrogen therapy that directly targets the androgen receptor (AR). Although these therapies reduce tumor burden and AR activity, the cancer inevitably recurs within 18–30 months. An approach targeting the androgen–AR axis at different levels could, therefore, improve the efficacy of prostate cancer therapy. Inhibition of 5α-reductase is one such approach; however, the two largest trials to investigate the use of the 5α-reductase inhibitors (5ARIs) finasteride and dutasteride in patients with prostate cancer have shown that, although the incidence of cancer was reduced by 5ARI treatment, those cancers that were detected were more aggressive than in patients treated with placebo. Thus, the best practice for using these drugs to prevent and treat prostate cancer remains unclear. PMID:21629218

  1. Targeting 5α-reductase for prostate cancer prevention and treatment.

    PubMed

    Nacusi, Lucas P; Tindall, Donald J

    2011-07-01

    Testosterone is the most abundant circulating androgen, and can be converted to dihydrotestosterone (DHT), a more potent androgen, by the 5α-reductase enzymes in target tissues. Current treatments for prostate cancer consist of reducing androgen levels by chemical or surgical castration or pure antiandrogen therapy that directly targets the androgen receptor (AR). Although these therapies reduce tumor burden and AR activity, the cancer inevitably recurs within 18-30 months. An approach targeting the androgen-AR axis at different levels could, therefore, improve the efficacy of prostate cancer therapy. Inhibition of 5α-reductase is one such approach; however, the two largest trials to investigate the use of the 5α-reductase inhibitors (5ARIs) finasteride and dutasteride in patients with prostate cancer have shown that, although the incidence of cancer was reduced by 5ARI treatment, those cancers that were detected were more aggressive than in patients treated with placebo. Thus, the best practice for using these drugs to prevent and treat prostate cancer remains unclear. PMID:21629218

  2. 40Ar/36Ar analyses of historic lava flows

    USGS Publications Warehouse

    Dalrymple, G.B.

    1969-01-01

    The ratio 40Ar/36Ar was measured for 26 subaerial historic lava flows. Approximately one-third of the samples had 40Ar/36Ar ratios either higher or lower than the atmospheric value of 295.5 at the 95% confidence level. Excess radiogenic 40Ar in five flows ranged from about 1 ?? 10-13 to 1.5 ?? 10-12 mol/g. Possible excess 36Ar in three flows was on the order of 10-16 to 10-15 mol/g. Upper 95% confidence limits for excess 40Ar in samples with normal 40Ar/36Ar ratios are generally less than 3 ?? 10-13 mol/g. The origin of the excess 36Ar is unknown but it may be due either to the incorporation of primitive argon that has been stored in the mantle in very low potassium environments or to enrichment in 36Ar as atmospheric argon diffuses into the rocks after they cool. ?? 1969.

  3. Evidence that biliverdin-IX beta reductase and flavin reductase are identical.

    PubMed Central

    Shalloe, F; Elliott, G; Ennis, O; Mantle, T J

    1996-01-01

    A search of the database shows that human biliverdin-IX beta reductase and flavin reductase are identical. We have isolated flavin reductase from bovine erythrocytes and show that the activity co-elutes with biliverdin-IX beta reductase. Preparations of the enzyme that are electrophoretically homogeneous exhibit both flavin reductase and biliverdin-IX beta reductase activities; however, they are not capable of catalysing the reduction of biliverdin-IX alpha. Although there is little obvious sequence identity between biliverdin-IX alpha reductase (BVR-A) and biliverdin-IX beta reductase (BVR-B), they do show weak immunological cross-reactivity. Both enzymes bind to 2',5'-ADP-Sepharose. PMID:8687377

  4. Fatty acyl-CoA reductase

    SciTech Connect

    Reiser, Steven E.; Somerville, Chris R.

    1998-12-01

    The present invention relates to bacterial enzymes, in particular to an acyl-CoA reductase and a gene encoding an acyl-CoA reductase, the amino acid and nucleic acid sequences corresponding to the reductase polypeptide and gene, respectively, and to methods of obtaining such enzymes, amino acid sequences and nucleic acid sequences. The invention also relates to the use of such sequences to provide transgenic host cells capable of producing fatty alcohols and fatty aldehydes.

  5. Synthesis and biological evaluation of novel gigantol derivatives as potential agents in prevention of diabetic cataract

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As a continuation of our efforts directed towards the development of natural anti-diabetic cataract agents, gigantol was isolated from Herba dendrobii and was found to inhibit both aldose reductase (AR) and inducible nitric oxide synthase (iNOS) activity, which play a significant role in the develop...

  6. Inhibitory Activities of Phenolic Compounds Isolated from Adina rubella Leaves Against 5α-Reductase Associated with Benign Prostatic Hypertrophy.

    PubMed

    Yin, Jun; Heo, Jun Hyeok; Hwang, Yoon Jeong; Le, Thi Tam; Lee, Min Won

    2016-01-01

    Adina rubella Hance (AR), a plant native to Korea, has been used as traditional medicine for dysentery, eczema, intoxication, and external hemorrhages. Previous phytochemical studies of AR have reported several components, including terpenoids, phenolics, and alkaloids. The current study evaluated the anti-oxidative and anti-inflammatory activities and 5α-reductase inhibition of isolated compounds of AR leaves to find a potential therapeutic agent for benign prostatic hypertrophy (BPH). Repeated chromatographic isolation of an 80% acetone extract of AR leaves yielded seven phenolic compounds: caffeic acid (1), chlorogenic acid (2), methyl chlorogenate (3), quercetin-3-rutinoside (4), kaempferol-3-O-α-l-rhamnopyranosyl-(1→6)-β-d-glucopyranoside (5), hyperoside (6), and grandifloroside (7). Compound 7 is a novel compound in AR. Caffeoyl derivatives 1-3 and 7 showed good anti-oxidative activities. In particular, caffeic acid (1) and grandifloroside (7) showed potent anti-inflammatory activities, and 7 also exhibited potent inhibitory activity against TNF-α and 5α-reductase. Our results show that the extract and grandifloroside (7) from leaves of AR might be developed as a source of potent anti-oxidative and anti-inflammatory agents and therapeutic agent for BPH. PMID:27399661

  7. Nitrate Reductase Regulates Expression of Nitrite Uptake and Nitrite Reductase Activities in Chlamydomonas reinhardtii 1

    PubMed Central

    Galván, Aurora; Cárdenas, Jacobo; Fernández, Emilio

    1992-01-01

    In Chlamydomonas reinhardtii mutants defective at the structural locus for nitrate reductase (nit-1) or at loci for biosynthesis of the molybdopterin cofactor (nit-3, nit-4, or nit-5 and nit-6), both nitrite uptake and nitrite reductase activities were repressed in ammonium-grown cells and expressed at high amounts in nitrogen-free media or in media containing nitrate or nitrite. In contrast, wild-type cells required nitrate induction for expression of high levels of both activities. In mutants defective at the regulatory locus for nitrate reductase (nit-2), very low levels of nitrite uptake and nitrite reductase activities were expressed even in the presence of nitrate or nitrite. Both restoration of nitrate reductase activity in mutants defective at nit-1, nit-3, and nit-4 by isolating diploid strains among them and transformation of a structural mutant upon integration of the wild-type nit-1 gene gave rise to the wild-type expression pattern for nitrite uptake and nitrite reductase activities. Conversely, inactivation of nitrate reductase by tungstate treatment in nitrate, nitrite, or nitrogen-free media made wild-type cells respond like nitrate reductase-deficient mutants with respect to the expression of nitrite uptake and nitrite reductase activities. Our results indicate that nit-2 is a regulatory locus for both the nitrite uptake system and nitrite reductase, and that the nitrate reductase enzyme plays an important role in the regulation of the expression of both enzyme activities. PMID:16668656

  8. Ar-39-Ar-40 ages of four ureilites

    NASA Technical Reports Server (NTRS)

    Bogard, D. D.; Garrison, D. H.

    1994-01-01

    Ureilites Novo Urei, Havero, and Kenna show strong evidence of one or more Ar-40 degassing events in the time period of 3.3-4.1 Ga ago. These ages may be compared to current interpretations of ureilite chronology. These include the suggestion of metasomatic activity on the parent body 3.7 Ga ago that reset some Sm-Nd ages and the suggestion that ureilites have experienced terrestrial contamination of several trace elements (including Pb and LREE), which makes suspect ages younger than approximately 4.5 Ga. Because the K-Ar chronometer can be sensitive to metamorphic events, we made Ar-39-Ar-40 determinations on bulk samples (0.12-0.14 g each) of four ureilites. The Ar-39-Ar-40 age spectra and K/Ca ratios as a function of cumulative Ar release from stepwise temperature extractions for the four ureilites analyzed are shown. Because Ar-39-Ar-40 ages shown by low and high temperature extractions may be suspect, we examined the intermediate temperature extractions. Although interpretation of these spectra is obviously uncertain, we believe that the most recent times of Ar degassing can be roughly inferred. These times are approximately 3.3 Ga for Havero, 3.3-3.7 Ga for Novo Urei, and approximately 4.1 Ga for Kenna, for which Ar degassing may not have been complete. The indication of Ar-39-Ar-40 degassing ages of 3.3-4.1 Ga for three ureilites that also contain an enhanced LREE component and (excepting Havero) produce a 3.74 Ga Sm-Nd age, suggests that both chronometers may have responded to the same parent body event. On the other hand, it is also possible that the Ar data reflect one or more separate events that did not strongly affect the Sm-Nd system, a situation that commonly occurs in eucrites. Thus the existence of reset Ar ages does not require similarly reset Sm-Nd ages.

  9. AR-39-AR-40 "Age" of Basaltic Shergottite NWA-3171

    NASA Technical Reports Server (NTRS)

    Bogard, Donald D.; Park, Jisun

    2007-01-01

    North-West-Africa 3171 is a 506 g, relatively fresh appearing, basaltic shergottite with similarities to Zagami and Shergotty, but not obviously paired with any of the other known African basaltic shergottites. Its exposure age has the range of 2.5-3.1 Myr , similar to those of Zagami and Shergotty. We made AR-39-AR-40 analyses of a "plagioclase" (now shock-converted to maskelynite) separate and of a glass hand-picked from a vein connected to shock melt pockets.. Plagioclase was separated using its low magnetic susceptibility and then heavy liquid with density of <2.85 g/cm(exp 3). The AR-39-AR-40 age spectrum of NWA-317 1 plag displays a rise in age over 20-100% of the 39Ar release, from 0.24 Gyr to 0.27 Gy.

  10. A metrological approach to measuring 40Ar* concentrations in K-Ar and 40Ar/39Ar mineral standards

    NASA Astrophysics Data System (ADS)

    Morgan, Leah E.; Postma, Onno; Kuiper, Klaudia F.; Mark, Darren F.; van der Plas, Wim; Davidson, Stuart; Perkin, Michael; Villa, Igor M.; Wijbrans, Jan R.

    2011-10-01

    In geochronology, isotopic ages are determined from the ratio of parent and daughter nuclide concentrations in minerals. For dating of geological material using the K-Ar system, the simultaneous determination of 40Ar and 40K concentrations on the same aliquot is not possible. Therefore, a widely used variant, the 40Ar/39Ar technique, involves the production of 39Ar from 39K by neutron bombardment and the reliance on indirect natural calibrators of the neutron flux, referred to as "mineral standards." Many mineral standards still in use rely on decades-old determinations of 40Ar concentrations; resulting uncertainties, both systematic and analytical, impede the determination of higher accuracy ages using the K-Ar decay system. We discuss the theoretical approach and technical design of a gas delivery system which emits metrologically traceable amounts of 40Ar and will allow for the sensitivity calibration of noble gas mass spectrometers. The design of this system is based on a rigorous assessment of the uncertainty budget and detailed tests of a prototype system. A number of obstacles and proposed resolutions are discussed along with the selection of components and their integration into a pipette system.

  11. Biliverdin reductase isozymes in metabolism.

    PubMed

    O'Brien, Luke; Hosick, Peter A; John, Kezia; Stec, David E; Hinds, Terry D

    2015-04-01

    The biliverdin reductase (BVR) isozymes BVRA and BVRB are cell surface membrane receptors with pleiotropic functions. This review compares, for the first time, the structural and functional differences between the isozymes. They reduce biliverdin, a byproduct of heme catabolism, to bilirubin, display kinase activity, and BVRA, but not BVRB, can act as a transcription factor. The binding motifs present in the BVR isozymes allow a wide range of interactions with components of metabolically important signaling pathways such as the insulin receptor kinase cascades, protein kinases (PKs), and inflammatory mediators. In addition, serum bilirubin levels have been negatively associated with abdominal obesity and hypertriglyceridemia. We discuss the roles of the BVR isozymes in metabolism and their potential as therapeutic targets. PMID:25726384

  12. An electrogenic nitric oxide reductase.

    PubMed

    Al-Attar, Sinan; de Vries, Simon

    2015-07-22

    Nitric oxide reductases (Nors) are members of the heme-copper oxidase superfamily that reduce nitric oxide (NO) to nitrous oxide (N₂O). In contrast to the proton-pumping cytochrome oxidases, Nors studied so far have neither been implicated in proton pumping nor have they been experimentally established as electrogenic. The copper-A-dependent Nor from Bacillus azotoformans uses cytochrome c₅₅₁ as electron donor but lacks menaquinol activity, in contrast to our earlier report (Suharti et al., 2001). Employing reduced phenazine ethosulfate (PESH) as electron donor, the main NO reduction pathway catalyzed by Cu(A)Nor reconstituted in liposomes involves transmembrane cycling of the PES radical. We show that Cu(A)Nor reconstituted in liposomes generates a proton electrochemical gradient across the membrane similar in magnitude to cytochrome aa₃, highlighting that bacilli using Cu(A)Nor can exploit NO reduction for increased cellular ATP production compared to organisms using cNor. PMID:26149211

  13. 5alpha-reductase: history and clinical importance.

    PubMed

    Marks, Leonard S

    2004-01-01

    The treatment of men with symptomatic benign prostatic hyperplasia (BPH) has shifted dramatically from surgery to drug therapy over the past decade. The revolution in BPH treatment began with the discovery of congenital 5alpha-reductase (5AR) deficiency, leading to the appreciation of 2 different androgenic hormones: testosterone, which mediates overt masculinization in the adult male, and dihydrotestosterone (DHT), which mediates prostatic growth, acne, facial beard, and male pattern baldness. Inhibition of DHT in adults results in prostatic shrinkage and symptomatic relief in many men, without the side effects seen with conventional androgen-deprivation therapy. The 5AR inhibitor drugs (finasteride and the dual inhibitor, dutasteride) are able to ablate the accumulation of intraprostatic DHT, the mechanism most responsible for prostate growth and maintenance. Not only may these drugs relieve symptoms, but they may also alter the natural history of the BPH process. Future indications for the 5ARI drugs could include chemoprevention of prostate cancer, prophylaxis of BPH-related complications, and treatment of BPH-associated hematuria. PMID:16985920

  14. 5 alpha-reductase deficiency without hypospadias.

    PubMed Central

    Ng, W K; Taylor, N F; Hughes, I A; Taylor, J; Ransley, P G; Grant, D B

    1990-01-01

    A boy aged 4 with penoscrotal hypospadias and his brother aged 12 with micropenis had typical changes of homozygous 5 alpha-reductase deficiency. After three injections of chorionic gonadotrophin there was a trivial rise in plasma dihydrotestosterone with a normal increase in plasma testosterone. Urine steroid chromatography showed abnormally high 5 beta: 5 alpha ratios and 5 alpha-reductase activity was appreciably reduced in genital skin fibroblasts. The results indicate that 5 alpha-reductase deficiency is not invariably associated with genital ambiguity. PMID:2248513

  15. Comparison of conventional K-Ar and 40Ar/39Ar dating of young mafic volcanic rocks

    USGS Publications Warehouse

    Lanphere, M.A.

    2000-01-01

    K-Ar and 40Ar/39Ar ages have been measured on nine mafic volcanic rocks younger than 1 myr from the Snake River Plain (Idaho), Mount Adams (Washington), and Crater Lake (Oregon). The K-Ar ages were calculated from Ar measurements made by isotope dilution and K2O measurements by flame photometry. The 40Ar/39Ar ages are incremental-heating experiments using a low-blank resistance-heated furnace. The results indicate that high-quality ages can be measured on young, mafic volcanic rocks using either the K-Ar or the 40Ar/39Ar technique. The precision of an 40Ar/39Ar plateau age generally is better than the precision of a K-Ar age because the plateau age is calculated by pooling the ages of several gas increments. The precision of a plateau age generally is better than the precision of an isotope correlation (isochron) age for the same sample. For one sample the intercept of the isochron yielded an 40Ar/36Ar value significantly different from the atmospheric value of 295.5. Recalculation of increment ages using the isochron intercept for the composition of nonradiogenic Ar in the sample resulted in much better agreement of ages for this sample. The results of this study also indicate that, given suitable material and modern equipment, precise K-Ar and 40Ar/39Ar ages can be measured on volcanic rocks as young as the latest Pleistocene, and perhaps even the Holocene.

  16. Ascorbate synthesis pathway, dual role of ascorbate in bone homeostasis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using mouse gene knock-out models, we identify aldehyde reductase (EC 1.1.1.2, Akr1a4 (GR)) and aldose reductase (EC 1.1.1.21, Akr1b3 (AR)) as the enzymes responsible for conversion of D-glucuronate to L-gulonate, a key step in the ascorbate (ASC) synthesis pathway in mice. The gene knock-out (KO) m...

  17. Genetics Home Reference: sepiapterin reductase deficiency

    MedlinePlus

    ... reductase enzyme. This enzyme is involved in the production of a molecule called tetrahydrobiopterin (also known as ... is responsible for the last step in the production of tetrahydrobiopterin. Tetrahydrobiopterin helps process several building blocks ...

  18. A dissimilatory nitrite reductase in Paracoccus halodenitrificans

    NASA Technical Reports Server (NTRS)

    Grant, M. A.; Hochstein, L. I.

    1984-01-01

    Paracoccus halodenitrificans produced a membrane-associated nitrite reductase. Spectrophotometric analysis showed it to be associated with a cd-cytochrome and located on the inner side of the cytoplasmic membrane. When supplied with nitrite, membrane preparations produced nitrous oxide and nitric oxide in different ratios depending on the electron donor employed. The nitrite reductase was maximally active at relatively low concentrations of sodium chloride and remained attached to the membranes at 100 mM sodium chloride.

  19. Neutron-hole states in 45Ar from 1H(46Ar, d) 45Ar reactions

    NASA Astrophysics Data System (ADS)

    Lu, F.; Lee, Jenny; Tsang, M. B.; Bazin, D.; Coupland, D.; Henzl, V.; Henzlova, D.; Kilburn, M.; Lynch, W. G.; Rogers, A. M.; Sanetullaev, A.; Sun, Z. Y.; Youngs, M.; Charity, R. J.; Sobotka, L. G.; Famiano, M.; Hudan, S.; Horoi, M.; Ye, Y. L.

    2013-07-01

    To improve the effective interactions in the pf shell, it is important to measure the single-particle and single-hole states near the N = 28 shell gap. In this paper, the neutron spectroscopic factors of hole states from the unstable neutron-rich 45Ar (Z = 18,N = 27) nucleus have been studied using the 1H(46Ar,d) 45Ar transfer reaction in inverse kinematics. Comparison of our results with the particle states of 45Ar produced in 2H(44Ar, p) 45Ar reaction shows that the two reactions populate states with different angular momenta. Using the angular distributions, we are able to confirm the spin assignments of four low-lying states of 45Ar. These are the ground state (f7/2), the first-excited state (p3/2), and the s1/2 and d3/2 states. While large basis shell-model predictions describe spectroscopic properties of the ground and p3/2 states very well, they fail to describe the s1/2 and d3/2 hole states.

  20. Pulsed discharge production Ar* metastables

    NASA Astrophysics Data System (ADS)

    Han, Jiande; Heaven, Michael C.; Emmons, Daniel; Perram, Glen P.; Weeks, David E.; Bailey, William F.

    2016-03-01

    The production of relatively high densities of Ar* metastables (>1012 cm-3) in Ar/He mixtures, at total pressures close to 1 atm, is essential for the efficient operation of an optically pumped Ar* laser. We have used emission spectroscopy and diode laser absorption spectroscopy measurements to observe the production and decay of Ar* in a parallel plate pulsed discharge. With discharge pulses of 1 μs duration we find that metastable production is dominated by processes occurring within the first 100 ns of the gas break-down. Application of multiple, closely spaced discharge pulses yields insights concerning conditions that favor metastable production. This information has been combined with time-resolved measurements of voltage and current. The experimental results and preliminary modeling of the discharge kinetics are presented.

  1. cAMP post-transcriptionally diminishes the abundance of adrenodoxin reductase mRNA.

    PubMed Central

    Brentano, S T; Black, S M; Lin, D; Miller, W L

    1992-01-01

    Adrenodoxin reductase (AR; ferridoxin: NADP+ oxidoreductase, EC 1.18.1.2) is a flavoprotein that mediates electron transport from NADPH to all known mitochondrial forms of cytochrome P450. AR mRNA was found in all human adult and fetal tissues examined; however, it was vastly more abundant in tissues that synthesize steroid hormones. The ratio of the 18- form of mRNA lacking 18 alternately spliced bases to the 18+ form was approximately 100:1 and remained constant irrespective of the tissue or hormonal manipulation, indicating that the alternate splicing is a passive nonregulated event. AR protein was unchanged by forskolin treatment of human JEG-3 cytotrophoblast cells for 24 h, but the mRNA diminished. Phorbol 12-myristate 13-acetate and cycloheximide had no effect, even though these agents had the expected effects on P450scc and adrenodoxin mRNAs. cAMP decreased the abundance of AR mRNA expressed from both transfected plasmids and the endogenous gene, indicating the effect was post-transcriptional. AR gene transcription in JEG-3 cells and promoter-chloramphenicol acetyltransferase constructs transfected into JEG-3 cells were unresponsive to forskolin. Powerful basal transcription elements were identified between -46 and -214 bases from the principal transcriptional initiation site, a region containing six elements closely resembling the binding site for transcription factor SP1. Images PMID:1315050

  2. Thioredoxin Reductase and its Inhibitors

    PubMed Central

    Saccoccia, Fulvio; Angelucci, Francesco; Boumis, Giovanna; Carotti, Daniela; Desiato, Gianni; Miele, Adriana E; Bellelli, Andrea

    2014-01-01

    Thioredoxin plays a crucial role in a wide number of physiological processes, which span from reduction of nucleotides to deoxyriboucleotides to the detoxification from xenobiotics, oxidants and radicals. The redox function of Thioredoxin is critically dependent on the enzyme Thioredoxin NADPH Reductase (TrxR). In view of its indirect involvement in the above mentioned physio/pathological processes, inhibition of TrxR is an important clinical goal. As a general rule, the affinities and mechanisms of binding of TrxR inhibitors to the target enzyme are known with scarce precision and conflicting results abound in the literature. A relevant analysis of published results as well as the experimental procedures is therefore needed, also in view of the critical interest of TrxR inhibitors. We review the inhibitors of TrxR and related flavoreductases and the classical treatment of reversible, competitive, non competitive and uncompetitive inhibition with respect to TrxR, and in some cases we are able to reconcile contradictory results generated by oversimplified data analysis. PMID:24875642

  3. Ar-39-Ar-40 Ages of Two Nakhlites, MIL03346 and Y000593: A Detailed Analysis

    NASA Technical Reports Server (NTRS)

    Park, Jisun; Garrison, Daniel; Bogard, Donald

    2007-01-01

    Radiometric dating of martian nakhlites by several techniques have given similar ages of approx.1.2-1.4 Ga [e.g. 1, 2]. Unlike the case with shergottites, where the presence of martian atmosphere and inherited radiogenic Ar-40 produce apparent Ar-39-Ar-40 ages older than other radiometric ages, Ar-Ar ages of nakhlites are similar to ages derived by other techniques. However, even in some nakhlites the presence of trapped martian Ar produces some uncertainty in the Ar-Ar age. We present here an analysis of such Ar-Ar ages from the MIL03346 and Y000593 nakhlites.

  4. An Astronomically Dated Standard in 40Ar/39Ar Geochronology?

    NASA Astrophysics Data System (ADS)

    Kuiper, K.; Hilgen, F.; Krijgsman, W.; Wijbrans, J.

    2003-12-01

    The standard geological time scale of Berggren et al. (1995) and Cande and Kent (1995) is calibrated with different absolute dating techniques, i.e. the Plio - Pleistocene relies on astronomical tuning, and older parts of the time scale are based on radio-isotopic (40Ar/39Ar and U/Pb) calibration methods. In the new edition of the standard geological timescale (Lourens et al., to be published in 2004) the entire Neogene will rely on astronomical dating. Therefore, it is of crucial importance that all dating methods produce equivalent absolute ages when the same geological event is dated. The Mediterranean Neogene provides an excellent opportunity to compare different dating methods by isotopic dating (40Ar/39Ar, U/Pb) of volcanic ash layers intercalated in astronomically dated sediments. Here we will show that in spite of potential errors in all methods, we succeeded to intercalibrate the 40Ar/39Ar and astronomical methods, arriving at astronomically calibrated age of 28.24 +/- 0.01 Ma for the in 40Ar/39Ar geochronology commonly used standard FCT sanidine. The advantage of an astronomically calibrated FCT above a K/Ar calibrated standard is a smaller error in the absolute age due to the lack of uncertainties related to 40K and radiogenic 40Ar contents in the primary standard and a decreasing influence of errors in the decay constant (branching ratio is not required). In addition to an astronomically calibrated FCT age we propose to introduce an astronomically dated standard. A direct astronomically dated standard can be regarded as a "primary" standard and does not require intercalibration with other standards, thus reducing analytical (and geological) uncertainties. Ash layers intercalated in sedimentary sequences in the Melilla Basin, Morocco appear to be the most suitable for this purpose. A reliable astronomical time control is available and intercalated ash layers contain sanidine phenocrysts up to 2 mm. Four ash layers are not or barely affected by

  5. Structural and mechanistic insights on nitrate reductases.

    PubMed

    Coelho, Catarina; Romão, Maria João

    2015-12-01

    Nitrate reductases (NR) belong to the DMSO reductase family of Mo-containing enzymes and perform key roles in the metabolism of the nitrogen cycle, reducing nitrate to nitrite. Due to variable cell location, structure and function, they have been divided into periplasmic (Nap), cytoplasmic, and membrane-bound (Nar) nitrate reductases. The first crystal structure obtained for a NR was that of the monomeric NapA from Desulfovibrio desulfuricans in 1999. Since then several new crystal structures were solved providing novel insights that led to the revision of the commonly accepted reaction mechanism for periplasmic nitrate reductases. The two crystal structures available for the NarGHI protein are from the same organism (Escherichia coli) and the combination with electrochemical and spectroscopic studies also lead to the proposal of a reaction mechanism for this group of enzymes. Here we present an overview on the current advances in structural and functional aspects of bacterial nitrate reductases, focusing on the mechanistic implications drawn from the crystallographic data. PMID:26362109

  6. Respiratory arsenate reductase as a bidirectional enzyme

    USGS Publications Warehouse

    Richey, C.; Chovanec, P.; Hoeft, S.E.; Oremland, R.S.; Basu, P.; Stolz, J.F.

    2009-01-01

    The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe–S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

  7. Respiratory arsenate reductase as a bidirectional enzyme

    SciTech Connect

    Richey, Christine; Chovanec, Peter; Hoeft, Shelley E.; Oremland, Ronald S.; Basu, Partha; Stolz, John F.

    2009-05-01

    The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe-S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

  8. Evolution of plant defense mechanisms. Relationships of phenylcoumaran benzylic ether reductases to pinoresinol-lariciresinol and isoflavone reductases.

    PubMed

    Gang, D R; Kasahara, H; Xia, Z Q; Vander Mijnsbrugge, K; Bauw, G; Boerjan, W; Van Montagu, M; Davin, L B; Lewis, N G

    1999-03-12

    Pinoresinol-lariciresinol and isoflavone reductase classes are phylogenetically related, as is a third, the so-called "isoflavone reductase homologs." This study establishes the first known catalytic function for the latter, as being able to engender the NADPH-dependent reduction of phenylcoumaran benzylic ethers. Accordingly, all three reductase classes are involved in the biosynthesis of important and related phenylpropanoid-derived plant defense compounds. In this investigation, the phenylcoumaran benzylic ether reductase from the gymnosperm, Pinus taeda, was cloned, with the recombinant protein heterologously expressed in Escherichia coli. The purified enzyme reduces the benzylic ether functionalities of both dehydrodiconiferyl alcohol and dihydrodehydrodiconiferyl alcohol, with a higher affinity for the former, as measured by apparent Km and Vmax values and observed kinetic 3H-isotope effects. It abstracts the 4R-hydride of the required NADPH cofactor in a manner analogous to that of the pinoresinol-lariciresinol reductases and isoflavone reductases. A similar catalytic function was observed for the corresponding recombinant reductase whose gene was cloned from the angiosperm, Populus trichocarpa. Interestingly, both pinoresinol-lariciresinol reductases and isoflavone reductases catalyze enantiospecific conversions, whereas the phenylcoumaran benzylic ether reductase only shows regiospecific discrimination. A possible evolutionary relationship among the three reductase classes is proposed, based on the supposition that phenylcoumaran benzylic ether reductases represent the progenitors of pinoresinol-lariciresinol and isoflavone reductases.

  9. Evaluation of nitrate reductase activity in Rhizobium japonicum

    SciTech Connect

    Streeter, J.G.; DeVine, P.J.

    1983-08-01

    Nitrate reductase activity was evaluated by four approaches, using four strains of Rhizobium japonicum and 11 chlorate-resistant mutants of the four strains. It was concluded that in vitro assays with bacteria or bacteroids provide the most simple and reliable assessment of the presence or absence of nitrate reductase. Nitrite reductase activity with methyl viologen and dithionite was found, but the enzyme activity does not confound the assay of nitrate reductase. 18 references

  10. 39Ar-40Ar "ages" and origin of excess 40Ar in Martian shergottites

    NASA Astrophysics Data System (ADS)

    Bogard, Donald; Park, Jisun; Garrison, Daniel

    2009-06-01

    We report new 39Ar-40Ar measurements on 15 plagioclase, pyroxene, and/or whole rock samples of 8 Martian shergottites. All age spectra suggest ages older than the meteorite formation ages, as defined by Sm-Nd and Rb-Sr isochrons. Employing isochron plots, only Los Angeles plagioclase and possibly Northwest Africa (NWA) 3171 plagioclase give ages in agreement with their formation ages. Isochrons for all shergottite samples reveal the presence of trapped Martian 40Ar (40Arxs), which exists in variable amounts in different lattice locations. Some 40Arxs is uniformly distributed throughout the lattice, resulting in a positive isochron intercept, and other 40Arxs occurs in association with K-bearing minerals and increases the isochron slope. These samples demonstrate situations where linear Ar isochrons give false ages that are too old. After subtracting 40Ar*that would accumulate by 40K decay since meteorite formation and small amounts of terrestrial 40Ar, all young age samples give similar 40Arxs concentrations of ˜1-2 × 10-6cm3/g, but a variation in K content by a factor of ˜80. Previously reported NASA Johnson Space Center data for Zagami, Shergotty, Yamato (Y-) 000097, Y-793605, and Queen Alexandra Range (QUE) 94201 shergottites show similar concentrations of 40Arxs to the new meteorite data reported here. Similar 40Arxs in different minerals and meteorites cannot be explained as arising from Martian atmosphere carried in strongly shocked phases such as melt veins. We invoke the explanation given by Bogard and Park (2008) for Zagami, that this 40Arxs in shergottites was acquired from the magma. Similarity in 40Arxs among shergottites may reveal common magma sources and/or similar magma generation and emplacement processes.

  11. Alteration of organic matter during infaunal polychaete gut passage and links to sediment organic geochemistry. Part II: Fatty acids and aldoses

    NASA Astrophysics Data System (ADS)

    Woulds, Clare; Middelburg, Jack J.; Cowie, Greg L.

    2014-07-01

    The activities of sediment-dwelling fauna are known to influence the rates of and pathways through which organic matter is cycled in marine sediments, and thus to influence eventual organic carbon burial or decay. However, due to methodological constraints, the role of faunal gut passage in determining the subsequent composition and thus degradability of organic matter is relatively little studied. Previous studies of organic matter digestion by benthic fauna have been unable to detect uptake and retention of specific biochemicals in faunal tissues, and have been of durations too short to fit digestion into the context of longer-term sedimentary degradation processes. Therefore this study aimed to investigate the aldose and fatty acid compositional alterations occurring to organic matter during gut passage by the abundant and ubiquitous polychaetes Hediste diversicolor and Arenicola marina, and to link these to longer-term changes typically observed during organic matter decay. This aim was approached through microcosm experiments in which selected polychaetes were fed with 13C-labelled algal detritus, and organisms, sediments, and faecal pellets were sampled at three timepoints over ∼6 weeks. Samples were analysed for their 13C-labelled aldose and fatty acid contents using GC-MS and GC-IRMS. Compound-selective net accumulation of biochemicals in polychaete tissues was observed for both aldoses and fatty acids, and the patterns of this were taxon-specific. The dominant patterns included an overall loss of glucose and polyunsaturated fatty acids; and preferential preservation or production of arabinose, microbial compounds (rhamnose, fucose and microbial fatty acids), and animal-synthesised fatty acids. These patterns may have been driven by fatty acid essentiality, preferential metabolism of glucose, and A. marina grazing on bacteria. Fatty acid suites in sediments from faunated microcosms showed greater proportions of saturated fatty acids and bacterial markers

  12. Structural prototypes for an extended family of flavoprotein reductases: comparison of phthalate dioxygenase reductase with ferredoxin reductase and ferredoxin.

    PubMed Central

    Correll, C. C.; Ludwig, M. L.; Bruns, C. M.; Karplus, P. A.

    1993-01-01

    The structure of phthalate dioxygenase reductase (PDR), a monomeric iron-sulfur flavoprotein that delivers electrons from NADH to phthalate dioxygenase, is compared to ferredoxin-NADP+ reductase (FNR) and ferredoxin, the proteins that reduce NADP+ in the final reaction of photosystem I. The folding patterns of the domains that bind flavin, NAD(P), and [2Fe-2S] are very similar in the two systems. Alignment of the X-ray structures of PDR and FNR substantiates the assignment of features that characterize a family of flavoprotein reductases whose members include cytochrome P-450 reductase, sulfite and nitrate reductases, and nitric oxide synthase. Hallmarks of this subfamily of flavoproteins, here termed the FNR family, are an antiparallel beta-barrel that binds the flavin prosthetic group, and a characteristic variant of the classic pyridine nucleotide-binding fold. Despite the similarities between FNR and PDR, attempts to model the structure of a dissociable FNR:ferredoxin complex by analogy with PDR reveal features that are at odds with chemical crosslinking studies (Zanetti, G., Morelli, D., Ronchi, S., Negri, A., Aliverti, A., & Curti, B., 1988, Biochemistry 27, 3753-3759). Differences in the binding sites for flavin and pyridine nucleotides determine the nucleotide specificities of FNR and PDR. The specificity of FNR for NADP+ arises primarily from substitutions in FNR that favor interactions with the 2' phosphate of NADP+. Variations in the conformation and sequences of the loop adjoining the flavin phosphate affect the selectivity for FAD versus FMN. The midpoint potentials for reduction of the flavin and [2Fe-2S] groups in PDR are higher than their counterparts in FNR and spinach ferredoxin, by about 120 mV and 260 mV, respectively. Comparisons of the structure of PDR with spinach FNR and with ferredoxin from Anabaena 7120, along with calculations of electrostatic potentials, suggest that local interactions, including hydrogen bonds, are the dominant

  13. Promiscuity and diversity in 3-ketosteroid reductases

    PubMed Central

    Penning, Trevor M.; Chen, Mo; Jin, Yi

    2014-01-01

    Many steroid hormones contain a Δ4-3-ketosteroid functionality that undergoes sequential reduction by 5α- or 5β- steroid reductases to produce 5α- or 5β-dihydrosteroids; and a subsequent 3-keto-reduction to produce a series of isomeric tetrahydrosteroids. Apart from steroid 5α-reductase all the remaining enzymes involved in the two step reduction process in humans belong to the aldo-keto reductase (AKR) superfamily. The enzymes involved in 3-ketosteroid reduction are AKR1C1–AKR1C4. These enzymes are promiscuous and also catalyze 20-keto- and 17-keto-steroid reduction. Interest in these reactions exist since they regulate steroid hormone metabolism in the liver, and in steroid target tissues, they may regulate steroid hormone receptor occupancy. In addition many of the dihydrosteroids are not biologically inert. The same enzymes are also involved in the metabolism of synthetic steroids e.g., hormone replacement therapeutics, contraceptive agents and inhaled glucocorticoids, and may regulate drug efficacy at their cognate receptors. This article reviews these reactions and the structural basis for substrate diversity in AKR1C1–AKR1C4, ketosteroid reductases. This article is part of a Special Issue entitled ‘Steroid/Sterol signaling’. PMID:25500069

  14. Promiscuity and diversity in 3-ketosteroid reductases.

    PubMed

    Penning, Trevor M; Chen, Mo; Jin, Yi

    2015-07-01

    Many steroid hormones contain a Δ(4)-3-ketosteroid functionality that undergoes sequential reduction by 5α- or 5β- steroid reductases to produce 5α- or 5β-dihydrosteroids; and a subsequent 3-keto-reduction to produce a series of isomeric tetrahydrosteroids. Apart from steroid 5α-reductase all the remaining enzymes involved in the two step reduction process in humans belong to the aldo-keto reductase (AKR) superfamily. The enzymes involved in 3-ketosteroid reduction are AKR1C1-AKR1C4. These enzymes are promiscuous and also catalyze 20-keto- and 17-keto-steroid reduction. Interest in these reactions exist since they regulate steroid hormone metabolism in the liver, and in steroid target tissues, they may regulate steroid hormone receptor occupancy. In addition many of the dihydrosteroids are not biologically inert. The same enzymes are also involved in the metabolism of synthetic steroids e.g., hormone replacement therapeutics, contraceptive agents and inhaled glucocorticoids, and may regulate drug efficacy at their cognate receptors. This article reviews these reactions and the structural basis for substrate diversity in AKR1C1-AKR1C4, ketosteroid reductases. This article is part of a Special Issue entitled 'Steroid/Sterol signaling'.

  15. Post-translational Regulation of Nitrate Reductase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nitrate reductase (NR) catalyzes the reduction of nitrate to nitrite, which is the first step in the nitrate assimilation pathway, but can also reduce nitrite to nitric oxide (NO), an important signaling molecule that is thought to mediate a wide array of of developmental and physiological processes...

  16. Synthesis of Nitrate Reductase in Chlorella

    PubMed Central

    Funkhouser, Edward A.; Shen, Teh-Chien; Ackermann, Renate

    1980-01-01

    Synthesis of nitrate reductase (EC 1.6.6.1) in Chlorella vulgaris was studied under inducing conditions, i.e. with cells grown on ammonia and then transferred to nitrate medium. Cycloheximide (but not chloramphenicol) completely inhibited synthesis of the enzyme, but only if it was added at the start (i.e. at the time of nitrate addition) of the induction period. Cycloheximide inhibition became less effective as induction by nitrate proceeded. Enzyme from small quantities of culture (1 to 3 milliliters of packed cells) was purified to homogeneity with the aid of blue dextran-Sepharose chromatography. Incorporation of radioactivity from labeled arginine into nitrate reductase was measured in the presence and absence of cycloheximide. Conditions were found under which the inhibitor completely blocked the incorporation of labeled amino acid, but only slightly decreased the increase in nitrate reductase activity. The results indicate that synthesis of nitrate reductase from amino acids proceeds by way of a protein precursor which is inactive enzymically. PMID:16661310

  17. Armenian Astronomical Society (ArAS) activities

    NASA Astrophysics Data System (ADS)

    Mickaelian, A. M.

    2016-09-01

    A review on the activities and achievements of Armenian Astronomical Society (ArAS) and Armenian astronomy in general during the last years is given. ArAS membership, ArAS electronic newsletters (ArASNews), ArAS webpage, Annual Meetings, Annual Prize for Young Astronomers (Yervant Terzian Prize) and other awards, international relations, presence in international organizations, local and international summer schools, science camps, astronomical Olympiads and other events, matters related to astronomical education, astronomical heritage, amateur astronomy, astronomy outreach and ArAS further projects are described and discussed.

  18. Production of ArN+ ions in the reactions Ar++N2 and N2++Ar

    NASA Astrophysics Data System (ADS)

    Tosi, Paolo; Correale, Raffaele; Lu, Wenyun; Bassi, Davide

    1999-03-01

    We have studied the production of ArN+ starting from the two symmetric charge-state reactants Ar++N2 and N2++Ar. For both reactions we measured the total cross sections as a function of the relative energy, in the energy range from 5 to 45 eV. Estimates of the reaction thresholds and symmetry considerations suggest that the reaction Ar++N2 produces ArN+ essentially in its first excited state A 3Π, while the reaction N2++Ar produces ground state ArN+ ions. This fact explains the discrepancies between different estimates of the dissociation energy of ArN+ obtained in previous studies.

  19. Resolvable miscalibration of the 40Ar/39Ar geochronometer

    NASA Astrophysics Data System (ADS)

    Mundil, R.; Renne, P. R.; Min, K. K.; Ludwig, K. R.

    2006-12-01

    U/Pb and 40Ar/39Ar isotopic dating techniques are the most widely applied geochronometers, both capable of 0.1% internal precision. A robust intercalibration between the two isotopic systems is fundamental for reconstructing short term processes and events in geologic time. However, whereas the U decay constants are known precisely (to ca 0.1%), the currently used 40K decay constant (5.543×10^{-10}/yr, (1)) is associated with an unstated uncertainty that is about an order of magnitude larger than the former, making high-resolution comparisons of ages from the two isotopic systems impossible. We present an indirect calibration by comparing radio-isotopic ages derived from both isotopic systems of rapidly cooled volcanic rocks in order to minimize effects from protracted cooling history. Eleven data pairs of 206Pb/238U and conventional 40Ar/39Ar ages exhibit a bias between the two isotopic systems ranging from >-1.5% for young rocks to ca -0.5% for rocks as old as 2 Ga (possibly even smaller for rocks >2 Ga), with the 40Ar/39Ar ages being consistently younger. All Mesozoic and Paleozoic samples display a bias of about -1%. Most of this bias is probably the result of miscalibration of the electron capture decay constant of 404→ 40Ar (λ40Kec) by ca -1%, in combination with a miscalibration of smaller magnitude and opposite sense of the β- decay constant (λ40Kβ-) of 40K→ 40Ca. Bias greater than 1% for younger Cenozoic samples probably reflects pre-eruptive zircon saturation (magma residence time) whose effects become proportionately negligible beyond ca. 200 Ma. Whereas the currently used decay constant for 40K (see above) is based on an arguably arbitrary selection from counting experiments associated with large and sometimes incomprehensible uncertainties (mostly from experiments conducted in the 1940s to 1960s) two recent recalibrations of λ40Ktotal using liquid scintillation counting techniques suggest precise and mutually consistent values of 5.553 ± 0

  20. Control of dihydrofolate reductase messenger ribonucleic acid production

    SciTech Connect

    Leys, E.J.; Kellems, R.E.

    1981-11-01

    The authors used methotrexate-resistant mouse cells in which dihydrofolate reductase levels are approximately 500 times normal to study the effect of growth stimulation on dihydrofolate reductase gene expression. As a result of growth stimulation, the relative rate of dihydrofolate reductase protein synthesis increased threefold, reaching a maximum between 25 and 30 h after stimulation. The relative rate of dihydrofolate reductase messenger ribonucleic acid production (i.e., the appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm) increased threefold after growth stimulation and was accompanied by a corresponding increase in the relative steady-state level of dihydrofolate reductase ribonucleic acid in the nucleus. However, the increase in the nuclear level of dihydrofolate reductase ribonucleic acid was not accompanied by a significant increase in the relative rate of transcription of the dihydrofolate reductase genes. These data indicated that the relative rate of appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm depends on the relative stability of the dihydrofolate reductase ribonucleic acid sequences in the nucleus and is not dependent on the relative rate of transcription of the dihydrofolate reductase genes.

  1. Augmentation of CFTR maturation by S-nitrosoglutathione reductase.

    PubMed

    Zaman, Khalequz; Sawczak, Victoria; Zaidi, Atiya; Butler, Maya; Bennett, Deric; Getsy, Paulina; Zeinomar, Maryam; Greenberg, Zivi; Forbes, Michael; Rehman, Shagufta; Jyothikumar, Vinod; DeRonde, Kim; Sattar, Abdus; Smith, Laura; Corey, Deborah; Straub, Adam; Sun, Fei; Palmer, Lisa; Periasamy, Ammasi; Randell, Scott; Kelley, Thomas J; Lewis, Stephen J; Gaston, Benjamin

    2016-02-01

    S-nitrosoglutathione (GSNO) reductase regulates novel endogenous S-nitrosothiol signaling pathways, and mice deficient in GSNO reductase are protected from airways hyperreactivity. S-nitrosothiols are present in the airway, and patients with cystic fibrosis (CF) tend to have low S-nitrosothiol levels that may be attributed to upregulation of GSNO reductase activity. The present study demonstrates that 1) GSNO reductase activity is increased in the cystic fibrosis bronchial epithelial (CFBE41o(-)) cells expressing mutant F508del-cystic fibrosis transmembrane regulator (CFTR) compared with the wild-type CFBE41o(-) cells, 2) GSNO reductase expression level is increased in the primary human bronchial epithelial cells expressing mutant F508del-CFTR compared with the wild-type cells, 3) GSNO reductase colocalizes with cochaperone Hsp70/Hsp90 organizing protein (Hop; Stip1) in human airway epithelial cells, 4) GSNO reductase knockdown with siRNA increases the expression and maturation of CFTR and decreases Stip1 expression in human airway epithelial cells, 5) increased levels of GSNO reductase cause a decrease in maturation of CFTR, and 6) a GSNO reductase inhibitor effectively reverses the effects of GSNO reductase on CFTR maturation. These studies provide a novel approach to define the subcellular location of the interactions between Stip1 and GSNO reductase and the role of S-nitrosothiols in these interactions.

  2. Retrieving the ars moriendi tradition.

    PubMed

    Leget, Carlo

    2007-09-01

    North Atlantic culture lacks a commonly shared view on dying well that helps the dying, their social environment and caregivers to determine their place and role, interpret death and deal with the process of ethical deliberation. What is lacking nowadays, however, has been part of Western culture in medieval times and was known as the ars moriendi (art of dying well) tradition. In this paper an updated version of this tradition is presented that meets the demands of present day secularized and multiform society. Five themes are central to the new art of dying: autonomy and the self, pain control and medical intervention, attachment and relations, life balance and guilt, death and afterlife. The importance of retrieving the ancient ars moriendi outreaches the boundaries of palliative medicine, since it deals with issues that play a central role in every context of medical intervention and treatment.

  3. Partial cloning, tissue distribution and effects of epigallocatechin gallate on hepatic 3-hydroxy-3-methylglutaryl-CoA reductase mRNA transcripts in goldfish (Carassius auratus).

    PubMed

    Cocci, Paolo; Mosconi, Gilberto; Palermo, Francesco Alessandro

    2014-07-25

    Epigallocatechin gallate (EGCG), the major active component of the green tea, has recently been found to inhibit 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCoAR) activity in vitro and to modulate lipogenesis in vivo. In this study we have evaluated the effects of short-term in vivo exposure to EGCG (6 μg g(-1) BW or 9 μg g(-1) BW) on hepatic HMGCoAR gene expression of goldfish (Carassius auratus). We initially characterized a partial sequence of goldfish HMGCoAR suggesting that the obtained fragment shares high similarity (>92%) with other fish HMGCoAR sequences. Further, the HMGCoAR transcript was detected in all goldfish tissues (except muscle) but primarily in liver, brain and gonads; on the contrary, low expression levels were found in intestine, heart, gill, and kidney. Both EGCG doses significantly decreased hepatic HMGCoAR mRNA levels 180 min post-injection. HMGCoAR was also significantly down-regulated at 90 min after injection in fish treated with the highest dose of EGCG. Our results demonstrate that hepatic HMGCoAR gene expression is acutely responsive to short-term EGCG exposure in goldfish. This finding suggests a potential role of EGCG in transcriptional regulation of the rate-limiting enzyme in cholesterol synthesis.

  4. arXiv.org and Physics Education

    ERIC Educational Resources Information Center

    Ramlo, Susan

    2007-01-01

    The website arXiv.org (pronounced "archive") is a free online resource for full-text articles in the fields of physics, mathematics, computer science, nonlinear science, and quantitative biology that has existed for about 15 years. Available directly at http://www.arXiv.org, this e-print archive is searchable. As of Jan. 3, 2007, arXiv had open…

  5. Synthesis and biological evaluation of 3-tetrazolo steroidal analogs: Novel class of 5α-reductase inhibitors.

    PubMed

    Aggarwal, Saurabh; Mahapatra, Manoj Kumar; Kumar, Rajnish; Bhardwaj, Tilak R; Hartmann, Rolf W; Haupenthal, Jörg; Kumar, Manoj

    2016-02-15

    In the present study, a series of steroidal tetrazole derivatives of androstane and pregnane have been prepared in which the tetrazole moiety was appended at C-3 and 17a-aza locations. 3-Tetrazolo-3,5-androstadien-17-one (6), 3-tetrazolo-19-nor-3,5-androstadien-17-one (10), 3-tetrazolo-3,5-pregnadien-20-one (14), 17a-substituted 3-tetrazolo-17a-aza-D-homo-3,5-androstadien-17-one (26-31) and 3-(2-acetyltetrazolo)-17a-aza-d-homo-3,5-androstadien-17-one (32) were synthesized from dehydroepiandrosterone acetate (1) through multiple synthetic steps. Some of the synthesized compounds were evaluated for their in vitro 5α-reductase (5AR) inhibitory activity by measuring the conversion of [(3)H] androstenedione in human embryonic kidney (HEK) cells. In vivo 5α-reductase inhibitory activity also showed a significant reduction (p <0.05) in rat prostate weight. The most potent compound 14 showed 5AR-2 inhibition with IC50 being 15.6nM as compared to clinically used drug finasteride (40nM). There was also a significant inhibition of 5AR-1 with IC50 547nM compared to finasteride (453nM). PMID:26780831

  6. Steroid 5α-reductase 2 deficiency.

    PubMed

    Mendonca, Berenice B; Batista, Rafael Loch; Domenice, Sorahia; Costa, Elaine M F; Arnhold, Ivo J P; Russell, David W; Wilson, Jean D

    2016-10-01

    Dihydrotestosterone is a potent androgen metabolite formed from testosterone by action of 5α-reductase isoenzymes. Mutations in the type 2 isoenzyme cause a disorder of 46,XY sex development, termed 5α-reductase type 2 deficiency and that was described forty years ago. Many mutations in the encoding gene have been reported in different ethnic groups. In affected 46,XY individuals, female external genitalia are common, but Mullerian ducts regress, and the internal urogenital tract is male. Most affected males are raised as females, but virilization occurs at puberty, and male social sex develops thereafter with high frequency. Fertility can be achieved in some affected males with assisted reproduction techniques, and adults with male social sex report a more satisfactory sex life and quality of life as compared to affected individuals with female social sex. PMID:27224879

  7. Discovery of pinoresinol reductase genes in sphingomonads.

    PubMed

    Fukuhara, Y; Kamimura, N; Nakajima, M; Hishiyama, S; Hara, H; Kasai, D; Tsuji, Y; Narita-Yamada, S; Nakamura, S; Katano, Y; Fujita, N; Katayama, Y; Fukuda, M; Kajita, S; Masai, E

    2013-01-10

    Bacterial genes for the degradation of major dilignols produced in lignifying xylem are expected to be useful tools for the structural modification of lignin in plants. For this purpose, we isolated pinZ involved in the conversion of pinoresinol from Sphingobium sp. strain SYK-6. pinZ showed 43-77% identity at amino acid level with bacterial NmrA-like proteins of unknown function, a subgroup of atypical short chain dehydrogenases/reductases, but revealed only 15-21% identity with plant pinoresinol/lariciresinol reductases. PinZ completely converted racemic pinoresinol to lariciresinol, showing a specific activity of 46±3 U/mg in the presence of NADPH at 30°C. In contrast, the activity for lariciresinol was negligible. This substrate preference is similar to a pinoresinol reductase, AtPrR1, of Arabidopsis thaliana; however, the specific activity of PinZ toward (±)-pinoresinol was significantly higher than that of AtPrR1. The role of pinZ and a pinZ ortholog of Novosphingobium aromaticivorans DSM 12444 were also characterized.

  8. Cinnamomi Cortex (Cinnamomum verum) Suppresses Testosterone-induced Benign Prostatic Hyperplasia by Regulating 5α-reductase

    PubMed Central

    Choi, Hyun-Myung; Jung, Yunu; Park, Jinbong; Kim, Hye-Lin; Youn, Dong-Hyun; Kang, JongWook; Jeong, Mi-Young; Lee, Jong-Hyun; Yang, Woong Mo; Lee, Seok-Geun; Ahn, Kwang Seok; Um, Jae-Young

    2016-01-01

    Cinnamomi cortex (dried bark of Cinnamomum verum) is an important drug in Traditional Korean Medicine used to improve blood circulation and Yang Qi. Benign prostatic hyperplasia (BPH) is a common chronic disease in aging men. This study was conducted to determine the effect of Cinnamomi cortex water extract (CC) on BPH. BPH was induced by a pre-4-week daily injection of testosterone propionate (TP). Six weeks of further injection with (a) vehicle, (b) TP, (c) TP + CC, (d) TP + finasteride (Fi) was carried on. As a result, the prostate weight and prostatic index of the CC treatment group were reduced. Histological changes including epithelial thickness and lumen area were recovered as normal by CC treatment. The protein expressions of prostate specific antigen, estrogen receptor α (ERα), androgen receptor (AR), 5α-reductase (5AR), and steroid receptor coactivator 1 were suppressed by treatment of CC. Immunohistochemical assays supported the western blot results, as the expressions of AR and ERα were down-regulated by CC treatment as well. Further in vitro experiments showed CC was able to inhibit proliferation of RWPE-1 cells by suppressing 5AR and AR. These results all together suggest CC as a potential treatment for BPH. PMID:27549514

  9. Cinnamomi Cortex (Cinnamomum verum) Suppresses Testosterone-induced Benign Prostatic Hyperplasia by Regulating 5α-reductase.

    PubMed

    Choi, Hyun-Myung; Jung, Yunu; Park, Jinbong; Kim, Hye-Lin; Youn, Dong-Hyun; Kang, JongWook; Jeong, Mi-Young; Lee, Jong-Hyun; Yang, Woong Mo; Lee, Seok-Geun; Ahn, Kwang Seok; Um, Jae-Young

    2016-01-01

    Cinnamomi cortex (dried bark of Cinnamomum verum) is an important drug in Traditional Korean Medicine used to improve blood circulation and Yang Qi. Benign prostatic hyperplasia (BPH) is a common chronic disease in aging men. This study was conducted to determine the effect of Cinnamomi cortex water extract (CC) on BPH. BPH was induced by a pre-4-week daily injection of testosterone propionate (TP). Six weeks of further injection with (a) vehicle, (b) TP, (c) TP + CC, (d) TP + finasteride (Fi) was carried on. As a result, the prostate weight and prostatic index of the CC treatment group were reduced. Histological changes including epithelial thickness and lumen area were recovered as normal by CC treatment. The protein expressions of prostate specific antigen, estrogen receptor α (ERα), androgen receptor (AR), 5α-reductase (5AR), and steroid receptor coactivator 1 were suppressed by treatment of CC. Immunohistochemical assays supported the western blot results, as the expressions of AR and ERα were down-regulated by CC treatment as well. Further in vitro experiments showed CC was able to inhibit proliferation of RWPE-1 cells by suppressing 5AR and AR. These results all together suggest CC as a potential treatment for BPH. PMID:27549514

  10. Carbohydrate utilization in Streptococcus thermophilus: characterization of the genes for aldose 1-epimerase (mutarotase) and UDPglucose 4-epimerase.

    PubMed Central

    Poolman, B; Royer, T J; Mainzer, S E; Schmidt, B F

    1990-01-01

    The complete nucleotide sequences of the genes encoding aldose 1-epimerase (mutarotase) (galM) and UDPglucose 4-epimerase (galE) and flanking regions of Streptococcus thermophilus have been determined. Both genes are located immediately upstream of the S. thermophilus lac operon. To facilitate the isolation of galE, a special polymerase chain reaction-based technique was used to amplify the region upstream of galM prior to cloning. The galM protein was homologous to the mutarotase of Acinetobacter calcoaceticus, whereas the galE protein was homologous to UDPglucose 4-epimerase of Escherichia coli and Streptomyces lividans. The amino acid sequences of galM and galE proteins also showed significant similarity with the carboxy-terminal and amino-terminal domains, respectively, of UDPglucose 4-epimerase from Kluyveromyces lactis and Saccharomyces cerevisiae, suggesting that the yeast enzymes contain an additional, yet unidentified (mutarotase) activity. In accordance with the open reading frames of the structural genes, galM and galE were expressed as polypeptides with apparent molecular masses of 39 and 37 kilodaltons, respectively. Significant activities of mutarotase and UDPglucose 4-epimerase were detected in lysates of E. coli cells containing plasmids encoding galM and galE. Expression of galE in E. coli was increased 300-fold when the gene was placed downstream of the tac promoter. The gene order for the gal-lac gene cluster of S. thermophilus is galE-galM-lacS-lacZ. The flanking regions of these genes were searched for consensus promoter sequences and further characterized by primer extension analysis. Analysis of mRNA levels for the gal and lac genes in S. thermophilus showed a strong reduction upon growth in medium containing glucose instead of lactose. The activities of the lac (lactose transport and beta-galactosidase) and gal (UDPglucose 4-epimerase) proteins of lactose- and glucose-grown S. thermophilus cells matched the mRNA levels. Images PMID:1694527

  11. Current status of 5α-reductase inhibitors in prostate disease management.

    PubMed

    Kang, Dong Il; Chung, Jae Il

    2013-04-01

    The key enzyme in the androgen synthesis and androgen receptor pathways is 5α-reductase (5-AR), which occurs as three isoenzymes. Types I and II 5-ARs the most important clinically, and two different 5-AR inhibitors (5-ARIs), finasteride and dutasteride, have been developed. Several urology associations have recommended and upgraded the use of 5-ARIs for an enlarged prostate with lower urinary tract symptoms. In the Prostate Cancer Prevention Trial and the Reduction by Dutasteride of Prostate Cancer Events Trial, 5-ARIs reduced the incidence of low-grade prostate cancer. However, despite the documented reductions in the overall incidence of prostate cancer, 5-ARIs are at the center of a dispute. The American Society of Clinical Oncology (ASCO) and the American Urology Association (AUA) presented clinical guidelines for the use of 5-ARIs for chemoprevention of prostate cancer in 2008. However, ASCO/AUA has eliminated these from the main "Clinical Guidelines" in 2012, because the U.S. Food and Drug Administration denied a supplemental New Drug Application for the use of dutasteride for prostate cancer chemoprevention. The 5-ARIs can also be used to manage hemospermia and prostatic hematuria, and to prevent intraoperative bleeding, although there is insufficient evidence for a standard strategy. This review summarizes the current use of 5-ARIs for prostate disease, including benign prostate hyperplasia, prostate cancer, prostate-related bleeding, and hemospermia. PMID:23614056

  12. Tumor-specific HMG-CoA reductase expression in primary premenopausal breast cancer predicts response to tamoxifen

    PubMed Central

    2011-01-01

    Introduction We previously reported an association between tumor-specific 3-hydroxy-3-methylglutharyl-coenzyme A reductase (HMG-CoAR) expression and a good prognosis in breast cancer. Here, the predictive value of HMG-CoAR expression in relation to tamoxifen response was examined. Methods HMG-CoAR protein and RNA expression was analyzed in a cell line model of tamoxifen resistance using western blotting and PCR. HMG-CoAR mRNA expression was examined in 155 tamoxifen-treated breast tumors obtained from a previously published gene expression study (Cohort I). HMG-CoAR protein expression was examined in 422 stage II premenopausal breast cancer patients, who had previously participated in a randomized control trial comparing 2 years of tamoxifen with no systemic adjuvant treatment (Cohort II). Kaplan-Meier analysis and Cox proportional hazards modeling were used to estimate the risk of recurrence-free survival (RFS) and the effect of HMG-CoAR expression on tamoxifen response. Results HMG-CoAR protein and RNA expression were decreased in tamoxifen-resistant MCF7-LCC9 cells compared with their tamoxifen-sensitive parental cell line. HMG-CoAR mRNA expression was decreased in tumors that recurred following tamoxifen treatment (P < 0.001) and was an independent predictor of RFS in Cohort I (hazard ratio = 0.63, P = 0.009). In Cohort II, adjuvant tamoxifen increased RFS in HMG-CoAR-positive tumors (P = 0.008). Multivariate Cox regression analysis demonstrated that HMG-CoAR was an independent predictor of improved RFS in Cohort II (hazard ratio = 0.67, P = 0.010), and subset analysis revealed that this was maintained in estrogen receptor (ER)-positive patients (hazard ratio = 0.65, P = 0.029). Multivariate interaction analysis demonstrated a difference in tamoxifen efficacy relative to HMG-CoAR expression (P = 0.05). Analysis of tamoxifen response revealed that patients with ER-positive/HMG-CoAR tumors had a significant response to tamoxifen (P = 0.010) as well as patients

  13. A Ferredoxin Disulfide Reductase Delivers Electrons to the Methanosarcina barkeri Class III Ribonucleotide Reductase.

    PubMed

    Wei, Yifeng; Li, Bin; Prakash, Divya; Ferry, James G; Elliott, Sean J; Stubbe, JoAnne

    2015-12-01

    Two subtypes of class III anaerobic ribonucleotide reductases (RNRs) studied so far couple the reduction of ribonucleotides to the oxidation of formate, or the oxidation of NADPH via thioredoxin and thioredoxin reductase. Certain methanogenic archaea contain a phylogenetically distinct third subtype of class III RNR, with distinct active-site residues. Here we report the cloning and recombinant expression of the Methanosarcina barkeri class III RNR and show that the electrons required for ribonucleotide reduction can be delivered by a [4Fe-4S] protein ferredoxin disulfide reductase, and a conserved thioredoxin-like protein NrdH present in the RNR operon. The diversity of class III RNRs reflects the diversity of electron carriers used in anaerobic metabolism.

  14. Role of the Dinitrogenase Reductase Arginine 101 Residue in Dinitrogenase Reductase ADP-Ribosyltransferase Binding, NAD Binding, and Cleavage

    PubMed Central

    Ma, Yan; Ludden, Paul W.

    2001-01-01

    Dinitrogenase reductase is posttranslationally regulated by dinitrogenase reductase ADP-ribosyltransferase (DRAT) via ADP-ribosylation of the arginine 101 residue in some bacteria. Rhodospirillum rubrum strains in which the arginine 101 of dinitrogenase reductase was replaced by tyrosine, phenylalanine, or leucine were constructed by site-directed mutagenesis of the nifH gene. The strain containing the R101F form of dinitrogenase reductase retains 91%, the strain containing the R101Y form retains 72%, and the strain containing the R101L form retains only 28% of in vivo nitrogenase activity of the strain containing the dinitrogenase reductase with arginine at position 101. In vivo acetylene reduction assays, immunoblotting with anti-dinitrogenase reductase antibody, and [adenylate-32P]NAD labeling experiments showed that no switch-off of nitrogenase activity occurred in any of the three mutants and no ADP-ribosylation of altered dinitrogenase reductases occurred either in vivo or in vitro. Altered dinitrogenase reductases from strains UR629 (R101Y) and UR630 (R101F) were purified to homogeneity. The R101F and R101Y forms of dinitrogenase reductase were able to form a complex with DRAT that could be chemically cross-linked by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide. The R101F form of dinitrogenase reductase and DRAT together were not able to cleave NAD. This suggests that arginine 101 is not critical for the binding of DRAT to dinitrogenase reductase but that the availability of arginine 101 is important for NAD cleavage. Both DRAT and dinitrogenase reductase can be labeled by [carbonyl-14C]NAD individually upon UV irradiation, but most 14C label is incorporated into DRAT when both proteins are present. The ability of R101F dinitrogenase reductase to be labeled by [carbonyl-14C]NAD suggested that Arg 101 is not absolutely required for NAD binding. PMID:11114923

  15. Part II. Evaluation of 40Ar- 39Ar quartz ages: Implications for fluid inclusion retentivity and determination of initial 40Ar/ 36Ar values in Proterozoic samples

    NASA Astrophysics Data System (ADS)

    Kendrick, M. A.; Miller, J. McL.; Phillips, D.

    2006-05-01

    The argon isotope systematics of vein-quartz samples with two different K-reservoirs have been evaluated in detail. Potassium is hosted by ultra-high-salinity fluid inclusions in quartz samples from the Eloise and Osborne iron-oxide-copper-gold (IOCG) deposits of the Mt Isa Inlier, Australia. In contrast, K is hosted by accidentally trapped mica within lower-salinity fluid inclusions of a sample selected from the Railway Fault, 13 km south of the Mt Isa copper mine, Australia. Imprecise apparent ages have been obtained for all of the samples studied and conclusively demonstrate that quartz fluid inclusions are retentive to Ar and have not leaked over billions of years. IOCG samples that host K in fluid inclusions only, have K/Cl values of <1 and the ages obtained represent the maximum ages for mineralization. In contrast, the Railway Fault samples that include accidentally trapped mica have K/Cl values of ≫1. Excess 40Ar E plus Cl hosted by fluid inclusions, and radiogenic 40Ar R plus K, are strongly correlated in these samples and define a plane in 3D 40Ar- 36Ar-K-Cl space. In this case, the plane yields an 'excess 40Ar E' corrected age of ˜1030 Ma that is 100's of Ma younger than nearby Cu-mineralization at Mt Isa. The age is interpreted to reflect 40Ar-loss from the accidentally trapped mica into the surrounding fluid inclusions, and is not related to the samples' age of formation. The initial 40Ar/ 36Ar value of fluid inclusions is widely used to provide information on fluid origin. For the IOCG samples that host K in fluid inclusions only, the initial 40Ar/ 36Ar values are close to the measured values at every temperature of stepped heating experiments. For samples that include accidentally trapped mica, the correction for post-entrapment radiogenic 40Ar R production is significant. Furthermore, because 39Ar K present in accidentally trapped mica crystals is released at different temperatures to radiogenic 40Ar R lost to the surrounding fluid inclusions

  16. Age measurements of potassium-bearing sulfide minerals by the 40Ar/39Ar technique

    USGS Publications Warehouse

    Czamanske, G.K.; Lanphere, M.A.; Erd, Richard C.; Blake, M.C., Jr.

    1978-01-01

    K-Ar ages have been determined for sulfide minerals for the first time. The occurrence of adequate amounts of potassium-bearing sulfides with ideal compositions K3Fe10S14 (???10 wt.% K) and KFe2S3 (???16 wt.% K) in samples from a mafic alkalic diatreme at Coyote Peak, California, prompted an attempt to date these materials. K3Fe10S14, a massive mineral with conchoidal fracture, gives an age of 29.4 ?? 0.5 m.y. (40Ar/39Ar), indistinguishable from the 28.3 ?? 0.4 m.y. (40Ar/39Ar) and 30.2 ?? 1.0 m.y.8 (conventional K-Ar) ages obtained for associated phlogopite (8.7 wt.% K). KFe2S3, a bladed, fibrous sulfide, gives a younger age, 26.5 ?? 0.5 m.y. (40Ar/39Ar), presumably owing to Ar loss. ?? 1978.

  17. Interpretation of discordant 40Ar/39Ar age-spectra of mesozoic tholeiites from antarctica

    USGS Publications Warehouse

    Fleck, R.J.; Sutter, J.F.; Elliot, D.H.

    1977-01-01

    Conventional K-Ar ages of tholeiitic basalts of the Ferrar Group in the central Transantarctic Mountains indicate significant loss of radiogenic 40Ar from this unit over much of its outcrop area. Argon loss varies inversely with amount of devitrified matrix in the basalts, which have not been thermally or tectonically disturbed since extrusion. 40Ar/19Ar age-spectra of these tholeiites are generally discordant and indicate significant inhomogeneity in the distribution of radiogenic 40Ar with respect to 39Ar, but are distinctly different from release patterns of thermally disturbed samples. Amounts of argon redistribution vary directly with amounts of devitrification and are reflected in progressive modification of the age spectra. A model of redistribution of radiogenic 40Ar by devitrification of originally glassy matrix is suggested that is consistent with disturbance of the conventional K-Ar systematics as well as the 40Ar/39Ar age-spectra. Samples with substantial redistribution but minor loss of radiogenic argon yield age spectra whose apparent ages decrease from low-temperature to high-temperature steps, similar to those reported for some lunar basalts, breccias, and soils. Modification of all the age spectra is attributed to redistribution of radiogenic 40Ar during progressive devitrification, although 39Ar-recoil effects suggested by Turner and Cadogan (1974) may be a factor in some cases. Where devitrification involves most potassium sites within the basalt, 40Ar/39Ar age-plateaux may be formed that have no geologic significance. ?? 1977.

  18. Ar-40-Ar-39 and Rb-Sr age determinations on Quaternary volcanic rocks

    NASA Technical Reports Server (NTRS)

    Radicati Di Brozolo, F.; Huneke, J. C.; Papanastassiou, D. A.; Wasserburg, G. J.

    1981-01-01

    Ages of leucite and biotite separates from samples of the potassic volcanics of the Roman Comagmatic region are derived by the stepwise degassing variant of the Ar-39-Ar-40 dating method and compared with those derived from Rb-Sr dating in order to evaluate the abilities of the methods to date Quaternary geological events. Six of the leucite separates are found to contain Ar with very high bulk 40/36 ratios and to have well correlated Ar-40 and Ar-39 contents, yielding ages of approximately 338,000 years. Two leucites observed to contain Ar with lower bulk 40/36 ratios and Ar-40/Ar-36 ratios significantly lower than atmospheric are found to have ages in substantial agreement with those of the other leucites despite the uncertainty in the composition of the trapped component. Ages obtained for the biotites are not as precise as those of the leucites, due to difficulties in obtaining a good separation of in situ radiogenic Ar-40 from trapped Ar-40. Ages determined from Rb-Sr measurements for selected tuff samples are found to be in good agreement with the Ar-40-Ar-39 ages of the leucites. Results demonstrate the possibility of attaining precisions of better than 5% in the dating of rocks 350,000 years old by both the Ar-40-Ar-39 and the Rb-Sr methods.

  19. A multiscale, mechanism-driven, dynamic model for the effects of 5α-reductase inhibition on prostate maintenance.

    PubMed

    Zager, Michael G; Barton, Hugh A

    2012-01-01

    A systems-level mathematical model is presented that describes the effects of inhibiting the enzyme 5α-reductase (5aR) on the ventral prostate of the adult male rat under chronic administration of the 5aR inhibitor, finasteride. 5aR is essential for androgen regulation in males, both in normal conditions and disease states. The hormone kinetics and downstream effects on reproductive organs associated with perturbing androgen regulation are complex and not necessarily intuitive. Inhibition of 5aR decreases the metabolism of testosterone (T) to the potent androgen 5α-dihydrotestosterone (DHT). This results in decreased cell proliferation, fluid production and 5aR expression as well as increased apoptosis in the ventral prostate. These regulatory changes collectively result in decreased prostate size and function, which can be beneficial to men suffering from benign prostatic hyperplasia (BPH) and could play a role in prostate cancer. There are two distinct isoforms of 5aR in male humans and rats, and thus developing a 5aR inhibitor is a challenging pursuit. Several inhibitors are on the market for treatment of BPH, including finasteride and dutasteride. In this effort, comparisons of simulated vs. experimental T and DHT levels and prostate size are depicted, demonstrating the model accurately described an approximate 77% decrease in prostate size and nearly complete depletion of prostatic DHT following 21 days of daily finasteride dosing in rats. This implies T alone is not capable of maintaining a normal prostate size. Further model analysis suggests the possibility of alternative dosing strategies resulting in similar or greater effects on prostate size, due to complex kinetics between T, DHT and gene occupancy. With appropriate scaling and parameterization for humans, this model provides a multiscale modeling platform for drug discovery teams to test and generate hypotheses about drugging strategies for indications like BPH and prostate cancer, such as compound

  20. Metrologically-Calibrated 40Ar Concentrations and Ages of Mineral Standards Used in 40Ar/39Ar Geochronology

    NASA Astrophysics Data System (ADS)

    Morgan, L. E.; Davidheiser-Kroll, B.; Kuiper, K.; Wijbrans, J. R.; Mark, D. F.

    2015-12-01

    In geochronology, isotopic ages are determined from the ratio of parent and daughter nuclide concentrations in minerals. For dating of geological material using the K-Ar system, the simultaneous determination of 40Ar and 40K concentrations on the same aliquot is not possible. Therefore, a widely used variant, the 40Ar/39Ar technique, involves the production of 39Ar from 39K by neutron bombardment and the reliance on indirect natural calibrators of the neutron flux, referred to as "mineral standards." Many mineral standards still in use rely on decades-old determinations of 40Ar concentrations; resulting uncertainties, both systematic and analytical, impede the determination of higher accuracy ages using the K-Ar decay system. We present results for the 40Ar concentrations and ages of mineral standards determined based on a modern gas delivery system (Morgan et al. 2011), which delivers metrologically-traceable amounts of 40Ar and thus allows for the sensitivity calibration of noble gas mass spectrometers.

  1. Ockham's razor and selective androgen receptor modulators (SARMs): are we overlooking the role of 5alpha-reductase?

    PubMed

    Gao, Wenqing; Dalton, James T

    2007-02-01

    Selective Androgen Receptor Modulators (SARMs) are a novel class of AR ligands that possess tissue-selective pharmacological activities. SARMs of various chemical structures have been discovered and characterized, and lead compounds with much improved specificity for AR, in vivo pharmacokinetic profiles, and higher degree of tissue selectivity have entered clinical development, and are expected to dramatically expand the clinical applications of androgens. With the rapid progress in SARM discovery and increasing demand for mechanism-based drug design, more and more research efforts have been devoted to the mechanisms of action of the observed tissue selectivity of SARMs. There is increasing enthusiasm in adapting the molecular mechanisms of action from SERM research to the SARM field; however, is the SARM story really so complicated? The tissue-specific expression of 5alpha-reductase might provide a simple explanation for this puzzle.

  2. Structure and function of NADPH-cytochrome P450 reductase and nitric oxide synthase reductase domain

    SciTech Connect

    Iyanagi, Takashi . E-mail: iyanagi@spring8.or.jp

    2005-12-09

    NADPH-cytochrome P450 reductase (CPR) and the nitric oxide synthase (NOS) reductase domains are members of the FAD-FMN family of proteins. The FAD accepts two reducing equivalents from NADPH (dehydrogenase flavin) and FMN acts as a one-electron carrier (flavodoxin-type flavin) for the transfer from NADPH to the heme protein, in which the FMNH {sup {center_dot}}/FMNH{sub 2} couple donates electrons to cytochrome P450 at constant oxidation-reduction potential. Although the interflavin electron transfer between FAD and FMN is not strictly regulated in CPR, electron transfer is activated in neuronal NOS reductase domain upon binding calmodulin (CaM), in which the CaM-bound activated form can function by a similar mechanism to that of CPR. The oxygenated form and spin state of substrate-bound cytochrome P450 in perfused rat liver are also discussed in terms of stepwise one-electron transfer from CPR. This review provides a historical perspective of the microsomal mixed-function oxidases including CPR and P450. In addition, a new model for the redox-linked conformational changes during the catalytic cycle for both CPR and NOS reductase domain is also discussed.

  3. Androgen receptor (AR) in cardiovascular diseases.

    PubMed

    Huang, Chiung-Kuei; Lee, Soo Ok; Chang, Eugene; Pang, Haiyan; Chang, Chawnshang

    2016-04-01

    Cardiovascular diseases (CVDs) are still the highest leading cause of death worldwide. Several risk factors have been linked to CVDs, including smoking, diabetes, hyperlipidemia, and gender among others. Sex hormones, especially the androgen and its receptor, androgen receptor (AR), have been linked to many diseases with a clear gender difference. Here, we summarize the effects of androgen/AR on CVDs, including hypertension, stroke, atherosclerosis, abdominal aortic aneurysm (AAA), myocardial hypertrophy, and heart failure, as well as the metabolic syndrome/diabetes and their impacts on CVDs. Androgen/AR signaling exacerbates hypertension, and anti-androgens may suppress hypertension. Androgen/AR signaling plays dual roles in strokes, depending on different kinds of factors; however, generally males have a higher incidence of strokes than females. Androgen and AR differentially modulate atherosclerosis. Androgen deficiency causes elevated lipid accumulation to enhance atherosclerosis; however, targeting AR in selective cells without altering serum androgen levels would suppress atherosclerosis progression. Androgen/AR signaling is crucial in AAA development and progression, and targeting androgen/AR profoundly restricts AAA progression. Men have increased cardiac hypertrophy compared with age-matched women that may be due to androgens. Finally, androgen/AR plays important roles in contributing to obesity and insulin/leptin resistance to increase the metabolic syndrome.

  4. Radical scavengers as ribonucleotide reductase inhibitors.

    PubMed

    Basu, Arijit; Sinha, Barij Nayan

    2012-01-01

    This paper compiled all the previous reports on radical scavengers, an interesting class of ribonucleotide reductase inhibitors. We have highlighted three key research areas: chemical classification of radical scavengers, structural and functional aspects of the radical site, and progress in drug designing for radical scavengers. Under the chemical classification section, we have recorded the discovery of hydroxyurea followed by discussions on hydroxamic acids, amidoximes, hydroxyguanidines, and phenolic compounds. In the next section, we have compiled the structural information for the radical site obtained from different crystallographic and theoretical studies. Finally, we have included the reported ligand based and structure based drug-designing studies.

  5. 40Ar/39Ar Interlaboratory Calibration into the Holocene.

    NASA Astrophysics Data System (ADS)

    Heizler, M. T.; Jicha, B.; Koppers, A. A. P.; Miggins, D. P.

    2015-12-01

    Advances in 40Ar/39Ar analytical precision for very young rocks requires collaborative efforts amongst argon geochronology labs to demonstrate age reproducibility commensurate with high precision. NM Tech (NMT), the University of Wisconsin (UW) and Oregon State University (OSU) have each dated Quaternary flux monitor standard AC-2 sanidine (~1.185 Ma), a blind sanidine described as being 50-100 ka (BS) and sanidine from the Qixiangshan (QIX) flow (~10 ka), Changbaishan volcano, China. The samples were irradiated in a single package with FC-2 sanidine (28.201 Ma) as the flux monitor and the irradiated material was distributed amongst the labs. Heizler was present during analysis at both OSU and UW and Jicha attended OSU during analysis. Physical presence was key towards gaining understanding of individual protocols and prompted valuable discussions. Analyses were carried out on single crystals using total fusion and/or step heating approaches. Age agreement was achieved within 2s uncertainty that ranged between (0.03-0.3%, 0.13-0.37% and 1.8-2.6%) for AC-2, BS and QIX, respectively. Each lab found AC-2 to vary somewhat beyond a normal distribution and to yield an age relative to FC-2 of ~1.185 Ma that is ~1.3% (~5-10 sigma) lower than some published estimates. A key cause of the variation between this study and previous results may be variable gas pressure equilibration times between extraction line and mass spectrometer coupled with variable choices to estimate time zero by other laboratories. The majority of our efforts concentrated on the QIX sanidine where prior data obtained by our labs revealed a factor of two spread in age (~11 and 23 ka) based on experiments carried out by total fusion and bulk incremental heating. By conducting single crystal age spectrum analysis we were able to mitigate effects of melt inclusion hosted excess argon and xenocrystic contamination towards obtaining analytical agreement with apparent ages near 10 ka. However, philosophical

  6. A mediated glucose/oxygen enzymatic fuel cell based on printed carbon inks containing aldose dehydrogenase and laccase as anode and cathode.

    PubMed

    Jenkins, Peter; Tuurala, Saara; Vaari, Anu; Valkiainen, Matti; Smolander, Maria; Leech, Dónal

    2012-03-10

    Enzyme electrodes show great potential for many applications, as biosensors and more recently as anodes and cathodes in biocatalytic fuel cells for power generation. Enzymes have advantages over metal catalysts, as they provide high specificity and reaction rates, while operating under mild conditions. Here we report on studies related to development of mass-producible, completely enzymatic printed glucose/oxygen biofuel cells. The cells are based on filter paper coated with conducting carbon inks containing mediators and laccase, for reduction of oxygen, or aldose dehydrogenase, for oxidation of glucose. Mediator performance in these printed formats is compared to relative rate constants for the enzyme-mediator reaction in solution, for a range of anode and cathode mediators. The power output and stability of fuels cells using an acidophilic laccase isolated from Trametes hirsuta is greater, at pH 5, than that for cells based on Melanocarpus albomyces laccase, that shows optimal activity closer to neutral pH, at pH 6. Highest power output, although of limited stability, was observed for ThL/ABTS cathodes, providing a maximum power density of 3.5 μWcm(-2) at 0.34 V, when coupled to an ALDH glucose anode mediated by an osmium complex. The stability of cell voltage above a threshold of 200 mV under a moderate 75 kΩ load is used to benchmark printed fuel cell performance. Highest stability was obtained for a printed fuel cell using osmium complexes as mediators of glucose oxidation by aldose dehydrogenase, and oxygen reduction by T. hirsuta laccase, maintaining cell voltage above 200 mV for 137 h at pH 5. These results provide promising directions for further development of mass-producible, completely enzymatic, printed biofuel cells. PMID:22305173

  7. Ar-Ar Ages of Brachinite and Brachinite-Like Achondrites

    NASA Astrophysics Data System (ADS)

    Beard, S. P.; Swindle, T. D.; Isachsen, C.

    2016-08-01

    There are few chronology studies on brachinite and brachinite-like achondrites. This work presents the first data of the study of Ar-Ar on a suite of achondrites, including NWA 595, NWA 1500, and NWA 6077.

  8. First-principles calibration of 40Ar/39Ar mineral standards and complete extraction of 40Ar* from sanidine

    NASA Astrophysics Data System (ADS)

    Morgan, L. E.; Kuiper, K.; Mark, D.; Postma, O.; Villa, I. M.; Wijbrans, J. R.

    2010-12-01

    40Ar/39Ar geochronology relies on comparing argon isotopic data for unknowns to those for knowns. Mineral standards used as neutron fluence monitors must be dated by the K-Ar method (or at least referenced to a mineral of known K-Ar age). The commonly used age of 28.02 ± 0.28 Ma for the Fish Canyon sanidine (FCs) (Renne et al., 1998) is based upon measurements of radiogenic 40Ar in GA1550 biotite (McDougall and Roksandic, 1974), but underlying full data were not published (these measurements were never intended for use as an international standard), so uncertainties are difficult to assess. Recent developments by Kuiper et al. (2008) and Renne et al. (2010) are limited by their reliance on the accuracy of other systems. Modern technology should allow for more precise and accurate calibration of primary K-Ar and 40Ar/39Ar standards. From the ideal gas law, the number of moles of 40Ar in a system can be calculated from measurements of pressure, volume, and temperature. Thus we have designed and are proceeding to build a pipette system to introduce well-determined amounts of 40Ar into noble gas extraction lines and mass spectrometers. This system relies on components with calibrations traceable to SI unit prototypes, including a diaphragm pressure gauge (MKS Instruments), thermocouples, and a “slug” of an accurately determined volume to be inserted into the reservoir for volume determinations of the reservoir and pipette. The system will be renewable, with a lifetime of ca. 1 month for gas in the reservoir, and portable, to permit interlaboratory calibrations. The quantitative extraction of 40Ar* from the mineral standard is of highest importance; for sanidine standards this is complicated by high melt viscosity during heating. Experiments adding basaltic “zero age glass” (ZAG) to decrease melt viscosity are underway. This has previously been explored by McDowell (1983) with a resistance furnace, but has not been quantitatively addressed with laser heating

  9. 40Ar/39Ar age spectra of some undisturbed terrestrial samples

    USGS Publications Warehouse

    Brent, Dalrymple G.; Lanphere, M.A.

    1974-01-01

    40Ar/39Ar age spectra and 40Ar/36Ar vs 39Ar/36Ar isochrons were determined by incremental heating for 11 terrestrial rocks and minerals whose geology indicates that they represent essentially undisturbed systems. The samples include muscovite, biotite, hornblende, sanidine, plagioclase, dacite, diabase and basalt and range in age from 40 to 1700 m.y. For each sample, the 40Ar/39Ar ratios, corrected for atmospheric and neutron-generated argon isotopes, are the same for most of the gas fractions released and the age spectra, which show pronounced plateaus, thus are consistent with models previously proposed for undisturbed samples. Plateau ages and isochron ages calculated using plateau age fractions are concordant and appear to be meaningful estimates of the crystallization and cooling ages of these samples. Seemingly anomalous age spectrum points can be attributed entirely to small amounts of previously unrecognized argon loss and to gas fractions that contain too small (less than 2 per cent) a proportion of the 39Ar released to be geologically significant. The use of a quantitative abscissa for age spectrum diagrams is recommended so that the size of each gas fraction is readily apparent. Increments containing less than about 4-5 per cent of the total 39Ar released should be interpreted cautiously. Both the age spectrum and isochron methods of data reduction for incremental heating experiments are worthwhile, as each gives slightly different but complementary information about the sample from the same basic data. Use of a least-squares fit that allows for correlated errors is recommended for 40Ar/36Ar vs 39Ar/36Ar isochrons. The results indicate that the 40Ar/39Ar incremental heating technique can be used to distinguish disturbed from undisturbed rock and mineral systems and will be a valuable geochronological tool in geologically complex terranes. ?? 1994.

  10. Diffusion of sup 40 Ar and sup 39 Ar in irradiated orthoclase

    SciTech Connect

    Foland, K.A.; Xu, Yuping )

    1990-11-01

    The important concerns of whether neutron irradiation affects Ar diffusion behavior in minerals and whether the diffusivities of natural radiogenic {sup 40}Ar and induced {sup 39}Ar are identical are considered. Both issues are addressed with isothermal and incremental-heating experiments on natural, homogeneous orthoclase from Benson Mines which was subjected to irradiation similar to typical {sup 40}Ar/{sup 39}Ar measurements. Previous study of this feldspar shows that laboratory {sup 40}Ar loss occurs by volume diffusion with physical grain sizes as the effective transport dimensions following a single Arrhenius relation. Isothermal-heating experiments on irradiated feldspar show the same loss and apparent {sup 40}Ar diffusion coefficients, within uncertainty, as unirradiated sample. For these heating times and temperatures, the experiments indicatet that the defects accompanying irradiation have only very minor, if any, effects on Ar behavior with regard to both diffusion kinetics and effective transport domains and that {sup 39}Ar and {sup 40}Ar diffusivities do not differ radically. Step-heating experiments yield essential flat spectra but with minor yet significant apparent discordance which is the same for different grain sizes. The spectra show high ages for small, initial low-temperature fractions and a slight increase in age with progressive Ar release. While these variations are potentially explicable by sample heterogeneities, the preferred explanation is that they result from {sup 39}Ar recoil loss and a slightly higher diffusivity of {sup 39}Ar relative to {sup 40}Ar. The results have important implications for diffusion processes in feldspar and the application of the {sup 40}Ar/{sup 39}Ar method.

  11. Re-Evaluation of Ar-39 - Ar-40 Ages for Apollo Lunar Rocks 15415 and 60015

    NASA Technical Reports Server (NTRS)

    Park, J.; Nyquist, L. E.; Bogard, D. D.; Garrison, D. H.; Shih, C.-Y.

    2010-01-01

    We re-analyzed 39Ar-40Ar ages of Apollo lunar highland samples 15415 and 60015, two ferroan anorthosites analyzed previously in the 1970 s, with a more detailed approach and with revised decay constants. From these samples we carefully prepared 100-200 mesh mineral separates for analysis at the Noble Gas Laboratory at NASA-Johnson Space Center. The Ar-39-Ar-40 age spectra for 15415 yielded an age of 3851 +/- 38 Ma with 33-99% of Ar39 release, roughly in agreement with previously reported Ar-Ar ages. For 60015, we obtained an age of 3584 +/- 152 Ma in 23-98% of Ar39 release, also in agreement with previously reported Ar-Ar ages of approximately 3.5 Ga. Highland anorthosites like these are believed by many to be the original crust of the moon, formed by plagioclase floatation atop a magma ocean, however the Ar-Ar ages of 15415 and 60015 are considerably younger than lunar crust formation. By contrast, recently recovered lunar anorthosites such as Dhofar 489, Dhofar 908, and Yamato 86032 yield older Ar-Ar ages, up to 4.35 Ga, much closer to time of formation of the lunar crust. It follows that the Ar-Ar ages of the Apollo samples must have been reset by secondary heating, and that this heating affected highland anorthosites at both the Apollo 15 and Apollo 16 landing sites but did not affect lunar highland meteorites. One obvious consideration is that while the Apollo samples were collected from the near side of the moon, these lunar meteorites are thought to have originated from the lunar far side

  12. The role of 5-alpha reductase inhibitors in prostate pathophysiology: Is there an additional advantage to inhibition of type 1 isoenzyme?

    PubMed

    Goldenberg, Larry; So, Alan; Fleshner, Neil; Rendon, Ricardo; Drachenberg, Darrel; Elhilali, Mostafa

    2009-06-01

    Normal growth and function of the prostate are contingent on the reduction of testosterone to dihydrotestosterone (DHT) by 5-alpha reductase (5-AR) enzymes types 1 and 2. It has been theorized that an overabundance of DHT may be implicated in the pathogenesis of both benign prostatic hyperplasia (BPH) and prostate cancer. Inhibitors of 5-AR such as dutasteride and finasteride may therefore have an important role in the prevention and treatment of BPH and prostate cancer. Dutasteride provides greater suppression of DHT than finasteride, thereby underlying the hypothesis that inhibition of both type 1 and type 2 would provide correspondingly greater protection than inhibition of type 2 alone. We review the potential significance of the 5-AR inhibitors in reducing the risk of prostate cancer according to the basic biology of prostate disease. PMID:19543428

  13. Structure of an integral membrane sterol reductase from Methylomicrobium alcaliphilum

    PubMed Central

    Li, Xiaochun; Roberti, Rita; Blobel, Günter

    2014-01-01

    Sterols are essential biological molecules in the majority of life forms. Sterol reductases1 including Delta-14 sterol reductase (C14SR), 7-dehydrocholesterol reductase (DHCR7) and 24-dehydrocholesterol reductase (DHCR24) reduce specific carbon-carbon double bonds of the sterol moiety using a reducing cofactor during sterol biosynthesis. Lamin B Receptor2 (LBR), an integral inner nuclear membrane protein, also contains a functional C14SR domain. Here we report the crystal structure of a Delta-14 sterol reductase (maSR1) from the methanotrophic bacterium Methylomicrobium alcaliphilum 20Z, a homolog of human C14SR, LBR, and DHCR7, with the cofactor NADPH. The enzyme contains 10 transmembrane segments (TM). Its catalytic domain comprises the C-terminal half (containing TM6-10) and envelops two interconnected pockets, one of which faces the cytoplasm and houses NADPH, while the other one is accessible from the lipid bilayer. Comparison with a soluble steroid 5β-reductase structure3 suggests that the reducing end of NADPH meets the sterol substrate at the juncture of the two pockets. A sterol reductase activity assay proves maSR1 can reduce the double bond of a cholesterol biosynthetic intermediate demonstrating functional conservation to human C14SR. Therefore, our structure as a prototype of integral membrane sterol reductases provides molecular insight into mutations in DHCR7 and LBR for inborn human diseases. PMID:25307054

  14. Biliverdin reductase: a target for cancer therapy?

    PubMed Central

    Gibbs, Peter E. M.; Miralem, Tihomir; Maines, Mahin D.

    2015-01-01

    Biliverdin reductase (BVR) is a multifunctional protein that is the primary source of the potent antioxidant, bilirubin. BVR regulates activities/functions in the insulin/IGF-1/IRK/PI3K/MAPK pathways. Activation of certain kinases in these pathways is/are hallmark(s) of cancerous cells. The protein is a scaffold/bridge and intracellular transporter of kinases that regulate growth and proliferation of cells, including PKCs, ERK and Akt, and their targets including NF-κB, Elk1, HO-1, and iNOS. The scaffold and transport functions enable activated BVR to relocate from the cytosol to the nucleus or to the plasma membrane, depending on the activating stimulus. This enables the reductase to function in diverse signaling pathways. And, its expression at the transcript and protein levels are increased in human tumors and the infiltrating T-cells, monocytes and circulating lymphocytes, as well as the circulating and infiltrating macrophages. These functions suggest that the cytoprotective role of BVR may be permissive for cancer/tumor growth. In this review, we summarize the recent developments that define the pro-growth activities of BVR, particularly with respect to its input into the MAPK signaling pathway and present evidence that BVR-based peptides inhibit activation of protein kinases, including MEK, PKCδ, and ERK as well as downstream targets including Elk1 and iNOS, and thus offers a credible novel approach to reduce cancer cell proliferation. PMID:26089799

  15. Ageing of glutathione reductase in the lens.

    PubMed

    Zhang, W Z; Augusteyn, R C

    1994-07-01

    The distribution of glutathione reductase activity in concentric layers from the lens has been determined as a function of age for 16 species. Primate lenses have almost ten times the level of glutathione reductase found in other species. Comparison with the activity of hexokinase revealed that this is not due to a higher overall rate of metabolism in these lenses. By contrast, the higher activity found in bird and fish lenses reflects a higher metabolic activity in these tissues. In all species, a gradient of activity was observed with the highest specific activity in the outermost cortical fibres, decreasing to virtually no activity in the inner parts of the tissue. No alterations were found in this gradient with increasing age, other than an increase in the amount of nuclear tissue essentially devoid of activity. The maximum activity in the outer cortical fibres was the same, regardless of the age of the lens. The time taken, in different species, for the specific activity to decrease by half, was estimated from the rate of protein accumulation. This time was found to vary from a few days to several years, indicating that the decrease in activity is not due to ageing but rather, it is related to the maturation of fibre cells. These observations are discussed in terms of current concepts of lens ageing and cataract formation. PMID:7835401

  16. Ar-Ar Age of NWA-1460 and Evidence For Young Formation Ages of the Shergottites

    NASA Technical Reports Server (NTRS)

    Bogard, Donald D.; Park, Jisun

    2006-01-01

    Agreement of Ar-Ar, Sm-Nd, and Rb-Sr ages for NWA1460, and the inconsistency between a low shock-heating temperature for Zagami and the proposition that a 4.0 Gyr-old Zagami lost most of its Ar-40 are inconsistent with ancient formation ages for these shergottites, but are consistent with relatively young igneous formation ages.

  17. AR function in promoting metastatic prostate cancer

    PubMed Central

    Augello, Michael A.; Den, Robert B.

    2015-01-01

    Prostate cancer (PCa) remains a leading cause of cancer-related death in the USA. While localized lesions are effectively treated through radical prostatectomy and/or radiation therapy, treatment for metastatic disease leverages the addiction of these tumors on the androgen receptor (AR) signaling axis for growth and disease progression. Though initially effective, tumors resistant to AR-directed therapeutics ultimately arise (a stage of the disease known as castration-resistant prostate cancer) and are responsible for PCa-specific mortality. Importantly, an abundance of clinical and preclinical evidence strongly implicates AR signaling cascades in the development of metastatic disease in both early and late stages, and thus a concerted effort has been made to delineate the AR-specific programs that facilitate progression to metastatic PCa. A multitude of downstream AR targets as well as critical AR cofactors have been identified which impinge upon both the AR pathway as well as associated metastatic phenotypes. This review will highlight the functional significance of these pathways to disseminated disease and define the molecular underpinnings behind these unique, AR-driven, metastatic signatures. PMID:24425228

  18. USDA/ARS Organic Production Research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    For much of its history, USDA/ARS had little to do with research on organic agriculture, however research in organic systems has made considerable gains at the agency over the past decade. In the 1980's and 1990's, as the organic food industry was taking off, ARS researchers who wanted to serve orga...

  19. 244-AR Vault Interim Stabilization Project Plan

    SciTech Connect

    LANEY, T.

    2000-03-24

    The 244-AR Vault Facility, constructed between 1966 and 1968, was designed to provide lag storage and treatment for the Plutonium-Uranium Extraction Facility (PUREX) tank farm sludges. Tank farm personnel transferred the waste from the 244-AR Vault Facility to B Plant for recovery of cesium and strontium. B Plant personnel then transferred the treatment residuals back to the tank farms for storage of the sludge and liquids. The last process operations, which transferred waste supporting the cesium/strontium recovery mission, occurred in April 1978. After the final transfer in 1978, the 244-AR facility underwent a cleanout. However, 2,271 L (600 gal) of sludge were left in Tank 004AR from an earlier transfer from Tank 241-AX-104. When the cleanout was completed, the facility was placed in a standby status. The sludge had been transferred to Tank 004AR to support Pacific Northwest National Laboratory [PNNL] vitrification work. Documentation of waste transfers suggests that a portion of the sludge may have been moved from Tank 004AR to Tank 002AR in preparation for transfer back to the AX Tank Farm; however, quantities of the sludge that were moved to Tank 002AR from that transfer must be estimated.

  20. Revised error propagation of 40Ar/39Ar data, including covariances

    NASA Astrophysics Data System (ADS)

    Vermeesch, Pieter

    2015-12-01

    The main advantage of the 40Ar/39Ar method over conventional K-Ar dating is that it does not depend on any absolute abundance or concentration measurements, but only uses the relative ratios between five isotopes of the same element -argon- which can be measured with great precision on a noble gas mass spectrometer. The relative abundances of the argon isotopes are subject to a constant sum constraint, which imposes a covariant structure on the data: the relative amount of any of the five isotopes can always be obtained from that of the other four. Thus, the 40Ar/39Ar method is a classic example of a 'compositional data problem'. In addition to the constant sum constraint, covariances are introduced by a host of other processes, including data acquisition, blank correction, detector calibration, mass fractionation, decay correction, interference correction, atmospheric argon correction, interpolation of the irradiation parameter, and age calculation. The myriad of correlated errors arising during the data reduction are best handled by casting the 40Ar/39Ar data reduction protocol in a matrix form. The completely revised workflow presented in this paper is implemented in a new software platform, Ar-Ar_Redux, which takes raw mass spectrometer data as input and generates accurate 40Ar/39Ar ages and their (co-)variances as output. Ar-Ar_Redux accounts for all sources of analytical uncertainty, including those associated with decay constants and the air ratio. Knowing the covariance matrix of the ages removes the need to consider 'internal' and 'external' uncertainties separately when calculating (weighted) mean ages. Ar-Ar_Redux is built on the same principles as its sibling program in the U-Pb community (U-Pb_Redux), thus improving the intercomparability of the two methods with tangible benefits to the accuracy of the geologic time scale. The program can be downloaded free of charge from

  1. 40Ar/39Ar and K-Ar data bearing on the metamorphic and tectonic history of western New England.

    USGS Publications Warehouse

    Sutter, J.F.; Ratcliffe, N.M.; Mukasa, S.B.

    1985-01-01

    40Ar/39Ar ages of coexisting biotite and hornblende from Proterozoic Y gneisses of the Berkshire and Green Mt massifs, as well as 40Ar/39Ar and K/Ar mineral and whole-rock ages from Palaeozoic metamorphic rocks, suggest that the thermal peaks for the dominant metamorphic recrystallization in western New England occurred 465 + or - 5 m.y. (Taconian). 40Ar/39Ar age data from a poorly-defined terrain along the eastern strip of the area suggests that the area has been retrograded during a metamorphism that peaked at least 376 + or - 5 m.y. (Acadian). Available age and petrological data from western New England indicate the presence of at least three separate metamorphic-structure domains of Taconic age: 1) a small area of relict high-P and low-T metamorphism, 2) a broad area of normal Barrovian metamorphism from chlorite to garnet grade characterized by a gentle metamorphic gradient and, 3) a rather narrow belt of steep-gradient, Barrovian series metamorphic rocks. Areas of maximum metamorphic intensity within the last domain coincide with areas of maximum crustal thickening in the later stage of Taconic orogeny. -L.di H

  2. ³⁹Ar/Ar measurements using ultra-low background proportional counters.

    PubMed

    Hall, Jeter; Aalseth, Craig E; Bonicalzi, Ricco M; Brandenberger, Jill M; Day, Anthony R; Humble, Paul H; Mace, Emily K; Panisko, Mark E; Seifert, Allen

    2016-01-01

    Age-dating groundwater and seawater using the (39)Ar/Ar ratio is an important tool to understand water mass-flow rates and mean residence time. Low-background proportional counters developed at Pacific Northwest National Laboratory use mixtures of argon and methane as counting gas. We demonstrate sensitivity to (39)Ar by comparing geological (ancient) argon recovered from a carbon dioxide gas well and commercial argon. The demonstrated sensitivity to the (39)Ar/Ar ratio is sufficient to date water masses as old as 1000 years. PMID:26516993

  3. ³⁹Ar/Ar measurements using ultra-low background proportional counters.

    PubMed

    Hall, Jeter; Aalseth, Craig E; Bonicalzi, Ricco M; Brandenberger, Jill M; Day, Anthony R; Humble, Paul H; Mace, Emily K; Panisko, Mark E; Seifert, Allen

    2016-01-01

    Age-dating groundwater and seawater using the (39)Ar/Ar ratio is an important tool to understand water mass-flow rates and mean residence time. Low-background proportional counters developed at Pacific Northwest National Laboratory use mixtures of argon and methane as counting gas. We demonstrate sensitivity to (39)Ar by comparing geological (ancient) argon recovered from a carbon dioxide gas well and commercial argon. The demonstrated sensitivity to the (39)Ar/Ar ratio is sufficient to date water masses as old as 1000 years.

  4. Spatial evaluation of Ar-systematics in rocks from the British Channel Islands: a UV laserprobe Ar/Ar study of excess 40Ar

    NASA Astrophysics Data System (ADS)

    Schwenzer, S. P.; Sherlock, S.; Kelley, S. P.

    2010-12-01

    The Ar-Ar method is a powerful tool for constraining thermal histories of metamorphic and plutonic rocks, most commonly undertaken on potassium rich mineral separates rather than whole rocks. While this approach usually yields reasonable thermal histories, it is rarely as precise as dating volcanics, and such rocks are also frequently contaminated by excess argon, artificially elevating the Ar-Ar ages. Understanding the evolution of excess argon represents a challenge; whilst it is possible to discern the ‘sink’ as the host mineral now contaminated by excess argon, examining the ‘source’ and ‘transport’ mechanism is more challenging. The approach we have taken here is to combine measurements of potassium rich and potassium poor minerals to understand the argon reservoirs and argon transfer between minerals, grain boundaries and fluids. Considering the system as a whole provides a method for understanding the complete history of the rock and thus assessing any interactions which may impact on the interpretation of ages and thermal history [Kelley, 2002, Chem. Geol. 188]. Here we have studied a series of plutonic and metamorphic basement samples from the British Channel Islands with different ages and post-emplacement histories, namely Icart Gneiss, Perelle Quartz Diorite, L’Ancresse Granodiorite, and Bordeaux Diorite. The formation age of Icart Gneiss is ~2000 Ma [D’Lemos et al., 1990, Geol. Soc. Spec. Pub. 51]. Ar-Ar ages of hornblendes and biotites from quartz diorites on Guernsey and Sark range between 606 and 596 Ma [Dallmeyer et al. 1991, J. Geol. Soc. London 148], whilst U-Pb zircon ages are in the range 710 to 613 Ma [Dallmeyer et al. 1991; Samson & D’Lemos 1998, J. Geol. Soc. London 155; Miller et al. 1999, Tectonophysics 132]. Detailed petrograpic (microscope, electron microprobe) investigations established the crystallization and deformation history of the samples, and revealed that post-magmatic alteration is unevenly distributed. This

  5. The aldo-keto reductases (AKRs): Overview.

    PubMed

    Penning, Trevor M

    2015-06-01

    The aldo-keto reductase (AKR) protein superfamily contains >190 members that fall into 16 families and are found in all phyla. These enzymes reduce carbonyl substrates such as: sugar aldehydes; keto-steroids, keto-prostaglandins, retinals, quinones, and lipid peroxidation by-products. Exceptions include the reduction of steroid double bonds catalyzed by AKR1D enzymes (5β-reductases); and the oxidation of proximate carcinogen trans-dihydrodiol polycyclic aromatic hydrocarbons; while the β-subunits of potassium gated ion channels (AKR6 family) control Kv channel opening. AKRs are usually 37kDa monomers, have an (α/β)8-barrel motif, display large loops at the back of the barrel which govern substrate specificity, and have a conserved cofactor binding domain. AKRs catalyze an ordered bi bi kinetic mechanism in which NAD(P)H cofactor binds first and leaves last. In enzymes that favor NADPH, the rate of release of NADP(+) is governed by a slow isomerization step which places an upper limit on kcat. AKRs retain a conserved catalytic tetrad consisting of Tyr55, Asp50, Lys84, and His117 (AKR1C9 numbering). There is conservation of the catalytic mechanism with short-chain dehydrogenases/reductases (SDRs) even though they show different protein folds. There are 15 human AKRs of these AKR1B1, AKR1C1-1C3, AKR1D1, and AKR1B10 have been implicated in diabetic complications, steroid hormone dependent malignancies, bile acid deficiency and defects in retinoic acid signaling, respectively. Inhibitor programs exist world-wide to target each of these enzymes to treat the aforementioned disorders. Inherited mutations in AKR1C and AKR1D1 enzymes are implicated in defects in the development of male genitalia and bile acid deficiency, respectively, and occur in evolutionarily conserved amino acids. The human AKRs have a large number of nsSNPs and splice variants, but in many instances functional genomics is lacking. AKRs and their variants are now poised to be interrogated using

  6. 40Ar/39Ar Ages of Carbonaceous Xenoliths in 2 HED Meteorites

    NASA Technical Reports Server (NTRS)

    Turrin, B.; Lindsay, F. N.; Park, J.; Herzog, G. F.; Delaney, J. S.; Swisher, C. C., III; Johnson, J.; Zolensky, M.

    2016-01-01

    The generally young K/Ar and 40Ar/39Ar ages of CM chondrites made us wonder whether carbonaceous xenoliths (CMX) entombed in Howardite–Eucrite–Diogenite (HED) meteorites might retain more radiogenic 40Ar than do ‘free-range’ CM-chondrites. To find out, we selected two HED breccias with carbonaceous inclusions in order to compare the 40Ar/39Ar release patterns and ages of the inclusions with those of nearby HED material. Carbonaceous inclusions (CMXs) in two HED meteorites lost a greater fraction of radiogenic 40Ar than did surrounding host material, but a smaller fraction of it than did free-range CM-chondrites such as Murchison or more heavily altered ones. Importantly, however, the siting of the CMXs in HED matrix did not prevent the 40Ar loss of about 40 percent of the radiogenic 40Ar, even from phases that degas at high laboratory temperatures. We infer that carbonaceous asteroids with perihelia of 1 astronomical unit probably experience losses of at least this size. The usefulness of 40Ar/39Ar dating for samples returned from C-type asteroids may hinge, therefore, on identifying and analyzing separately small quantities of the most retentive phases of carbonaceous chondrites.

  7. Docking and molecular dynamics studies at trypanothione reductase and glutathione reductase active sites.

    PubMed

    Iribarne, Federico; Paulino, Margot; Aguilera, Sara; Murphy, Miguel; Tapia, Orlando

    2002-05-01

    A theoretical docking study on the active sites of trypanothione reductase (TR) and glutathione reductase (GR) with the corresponding natural substrates, trypanothione disulfide (T[S]2) and glutathione disulfide (GSSG), is reported. Molecular dynamics simulations were carried out in order to check the robustness of the docking results. The energetic results are in agreement with previous experimental findings and show the crossed complexes have lower stabilization energies than the natural ones. To test DOCK3.5, four nitro furanic compounds, previously designed as potentially active anti-chagasic molecules, were docked at the GR and TR active sites with the DOCK3.5 procedure. A good correlation was found between differential inhibitory activity and relative interaction energy (affinity). The results provide a validation test for the use of DOCK3.5 in connection with the design of anti-chagasic drugs.

  8. Transcripts of anthocyanidin reductase and leucoanthocyanidin reductase and measurement of catechin and epicatechin in tartary buckwheat.

    PubMed

    Kim, Yeon Bok; Thwe, Aye Aye; Kim, Yeji; Li, Xiaohua; Cho, Jin Woong; Park, Phun Bum; Valan Arasu, Mariadhas; Abdullah Al-Dhabi, Naif; Kim, Sun-Ju; Suzuki, Tastsuro; Hyun Jho, Kwang; Park, Sang Un

    2014-01-01

    Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs) such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions.

  9. Transcripts of Anthocyanidin Reductase and Leucoanthocyanidin Reductase and Measurement of Catechin and Epicatechin in Tartary Buckwheat

    PubMed Central

    Kim, Yeon Bok; Thwe, Aye Aye; Kim, YeJi; Li, Xiaohua; Cho, Jin Woong; Park, Phun Bum; Valan Arasu, Mariadhas; Abdullah Al-Dhabi, Naif; Kim, Sun-Ju; Suzuki, Tastsuro; Hyun Jho, Kwang; Park, Sang Un

    2014-01-01

    Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs) such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions. PMID:24605062

  10. Transcripts of anthocyanidin reductase and leucoanthocyanidin reductase and measurement of catechin and epicatechin in tartary buckwheat.

    PubMed

    Kim, Yeon Bok; Thwe, Aye Aye; Kim, Yeji; Li, Xiaohua; Cho, Jin Woong; Park, Phun Bum; Valan Arasu, Mariadhas; Abdullah Al-Dhabi, Naif; Kim, Sun-Ju; Suzuki, Tastsuro; Hyun Jho, Kwang; Park, Sang Un

    2014-01-01

    Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) play an important role in the monomeric units biosynthesis of proanthocyanidins (PAs) such as catechin and epicatechin in several plants. The aim of this study was to clone ANR and LAR genes involved in PAs biosynthesis and examine the expression of these two genes in different organs under different growth conditions in two tartary buckwheat cultivars, Hokkai T8 and T10. Gene expression was carried out by quantitative real-time RT-PCR, and catechin and epicatechin content was analyzed by high performance liquid chromatography. The expression pattern of ANR and LAR did not match the accumulation pattern of PAs in different organs of two cultivars. Epicatechin content was the highest in the flowers of both cultivars and it was affected by light in only Hokkai T8 sprouts. ANR and LAR levels in tartary buckwheat might be regulated by different mechanisms for catechin and epicatechin biosynthesis under light and dark conditions. PMID:24605062

  11. Ar-40/Ar-39 laser-probe dating of diamond inclusions from the Premier kimberlite

    NASA Technical Reports Server (NTRS)

    Phillips, D.; Onstott, T. C.; Harris, J. W.

    1989-01-01

    The results of Ar-40/Ar-39 laser-probe analyses of individual eclogitic clinopyroxene inclusions from Premier diamonds are reported which yield a mean age of 1198 + or - 14 Myr. This age agrees well with Sm-Nd and Ar-40/Ar-39 analyses on similar Premier inclusions and is indistinguishable from the inferred time of emplacement of the host kimberlite, which implies that diamond formation was essentially synchronous with kimberlite generation. The extrapolated nonradiogenic Ar-40/Ar-36 ratio of 334 + or - 102 is similar to the present-day atmospheric composition. This value is inconsistent with Sr and Nd isotopic signatures from Premier eclogite inclusions, which suggest a depleted mantle source. Preentrapment equilibration of the inclusions with an Ar-36-rich fluid is the most probable explanation for the low nonradiogenic composition.

  12. Ribonucleotide reductases: essential enzymes for bacterial life

    PubMed Central

    Torrents, Eduard

    2014-01-01

    Ribonucleotide reductase (RNR) is a key enzyme that mediates the synthesis of deoxyribonucleotides, the DNA precursors, for DNA synthesis in every living cell. This enzyme converts ribonucleotides to deoxyribonucleotides, the building blocks for DNA replication, and repair. Clearly, RNR enzymes have contributed to the appearance of genetic material that exists today, being essential for the evolution of all organisms on Earth. The strict control of RNR activity and dNTP pool sizes is important, as pool imbalances increase mutation rates, replication anomalies, and genome instability. Thus, RNR activity should be finely regulated allosterically and at the transcriptional level. In this review we examine the distribution, the evolution, and the genetic regulation of bacterial RNRs. Moreover, this enzyme can be considered an ideal target for anti-proliferative compounds designed to inhibit cell replication in eukaryotic cells (cancer cells), parasites, viruses, and bacteria. PMID:24809024

  13. Ribonucleotide reductases: essential enzymes for bacterial life.

    PubMed

    Torrents, Eduard

    2014-01-01

    Ribonucleotide reductase (RNR) is a key enzyme that mediates the synthesis of deoxyribonucleotides, the DNA precursors, for DNA synthesis in every living cell. This enzyme converts ribonucleotides to deoxyribonucleotides, the building blocks for DNA replication, and repair. Clearly, RNR enzymes have contributed to the appearance of genetic material that exists today, being essential for the evolution of all organisms on Earth. The strict control of RNR activity and dNTP pool sizes is important, as pool imbalances increase mutation rates, replication anomalies, and genome instability. Thus, RNR activity should be finely regulated allosterically and at the transcriptional level. In this review we examine the distribution, the evolution, and the genetic regulation of bacterial RNRs. Moreover, this enzyme can be considered an ideal target for anti-proliferative compounds designed to inhibit cell replication in eukaryotic cells (cancer cells), parasites, viruses, and bacteria. PMID:24809024

  14. Ribonucleotide reductases: essential enzymes for bacterial life.

    PubMed

    Torrents, Eduard

    2014-01-01

    Ribonucleotide reductase (RNR) is a key enzyme that mediates the synthesis of deoxyribonucleotides, the DNA precursors, for DNA synthesis in every living cell. This enzyme converts ribonucleotides to deoxyribonucleotides, the building blocks for DNA replication, and repair. Clearly, RNR enzymes have contributed to the appearance of genetic material that exists today, being essential for the evolution of all organisms on Earth. The strict control of RNR activity and dNTP pool sizes is important, as pool imbalances increase mutation rates, replication anomalies, and genome instability. Thus, RNR activity should be finely regulated allosterically and at the transcriptional level. In this review we examine the distribution, the evolution, and the genetic regulation of bacterial RNRs. Moreover, this enzyme can be considered an ideal target for anti-proliferative compounds designed to inhibit cell replication in eukaryotic cells (cancer cells), parasites, viruses, and bacteria.

  15. Monodehydroascorbate reductase mediates TNT toxicity in plants.

    PubMed

    Johnston, Emily J; Rylott, Elizabeth L; Beynon, Emily; Lorenz, Astrid; Chechik, Victor; Bruce, Neil C

    2015-09-01

    The explosive 2,4,6-trinitrotoluene (TNT) is a highly toxic and persistent environmental pollutant. Due to the scale of affected areas, one of the most cost-effective and environmentally friendly means of removing explosives pollution could be the use of plants. However, mechanisms of TNT phytotoxicity have been elusive. Here, we reveal that phytotoxicity is caused by reduction of TNT in the mitochondria, forming a nitro radical that reacts with atmospheric oxygen, generating reactive superoxide. The reaction is catalyzed by monodehydroascorbate reductase 6 (MDHAR6), with Arabidopsis deficient in MDHAR6 displaying enhanced TNT tolerance. This discovery will contribute toward the remediation of contaminated sites. Moreover, in an environment of increasing herbicide resistance, with a shortage in new herbicide classes, our findings reveal MDHAR6 as a valuable plant-specific target.

  16. GRS Measurements of Ar in Mars' Atmosphere

    NASA Astrophysics Data System (ADS)

    Sprague, A. L.; Boynton, W. V.; Kerry, K. E.; Janes, D. M.; Kelly, N. J.; Crombie, M. K.; Hunten, D. M.; Nelli, S. M.; Murphy, J. R.; Reedy, R. C.; Metzger, A. E.

    2005-08-01

    One and one half Mars years of atmospheric argon (Ar) measurements are described in the context of understanding how Ar, a minor constituent of Mars atmosphere that does not condense at Mars temperatures, can be used to study martian circulation and dynamics. There is a repeated factor of 6 enhancement of Ar measured over south polar latitudes. The maximum in Ar abundance occurs near the onset of southern winter. There is no similar strong enhancement of Ar over north-polar regions during northern winter; only modest evidence for an enhancement peak is present. Part of this difference is explained by the global topographic dichotomy and the fact that the duration of northern autumn and winter is shorter than southern autumn and winter. Rapid seasonal fluctuations in Ar abundance may indicate evidence for wave activity at the perimeter of the southern seasonal polar cap. The apparent lack of coincidence of Ar enhancement with the relatively cold, cryptic terrain or relatively warm, bright albedo regions, indirectly supports the conclusion that the low temperatures measured over the south polar region by IRTM are probably caused by the combination of low CO2 abundance over south polar night and low emissivity regions on the surface associated with small grain size.

  17. arXiv.org and Physics Education

    NASA Astrophysics Data System (ADS)

    Ramlo, Susan

    2007-09-01

    The website arXiv.org (pronounced archive) is a free online resource for full-text articles in the fields of physics, mathematics, computer science, nonlinear science, and quantitative biology that has existed for about 15 years. Available directly at http://www.arXiv.org, this e-print archive is searchable. As of Jan. 3, 2007, arXiv had open access to 401,226 e-prints in the topic areas. Those who sign up for an ID and password can also sign up for daily submission abstract emails for specific subject classes of arXiv, including physics education, physics and society, and history of physics. Founded and developed by Paul Ginsparg when he was at Los Alamos National Laboratory, arXiv's original name was the LANL preprint archive or xxx.lanl.gov. The location and name changed after Ginsparg moved to the physics department at Cornell University. Today, arXiv is hosted and operated by Cornell University library. Mirror sites for arXiv exist worldwide.2

  18. The cytochrome bd respiratory oxygen reductases

    PubMed Central

    Borisov, Vitaliy B.; Gennis, Robert B.; Hemp, James; Verkhovsky, Michael I.

    2011-01-01

    Summary Cytochrome bd is a respiratory quinol:O2 oxidoreductase found in many prokaryotes, including a number of pathogens. The main bioenergetic function of the enzyme is the production of a proton motive force by the vectorial charge transfer of protons. The sequences of cytochromes bd are not homologous to those of the other respiratory oxygen reductases, i.e., the heme-copper oxygen reductases or alternative oxidases (AOX). Generally, cytochromes bd are noteworthy for their high affinity for O2 and resistance to inhibition by cyanide. In E. coli, for example, cytochrome bd (specifically, cytochrome bd-I) is expressed under O2-limited conditions. Among the members of the bd-family are the so-called cyanide-insensitive quinol oxidases (CIO) which often have a low content of the eponymous heme d but, instead, have heme b in place of heme d in at least a majority of the enzyme population. However, at this point, no sequence motif has been identified to distinguish cytochrome bd (with a stoichiometric complement of heme d) from an enzyme designated as CIO. Members of the bd-family can be subdivided into those which contain either a long or a short hydrophilic connection between transmembrane helices 6 and 7 in subunit I, designated as the Q-loop. However, it is not clear whether there is a functional consequence of this difference. This review summarizes current knowledge on the physiological functions, genetics, structural and catalytic properties of cytochromes bd. Included in this review are descriptions of the intermediates of the catalytic cycle, the proposed site for the reduction of O2, evidence for a proton channel connecting this active site to the bacterial cytoplasm, and the molecular mechanism by which a membrane potential is generated. PMID:21756872

  19. The cytochrome bd respiratory oxygen reductases.

    PubMed

    Borisov, Vitaliy B; Gennis, Robert B; Hemp, James; Verkhovsky, Michael I

    2011-11-01

    Cytochrome bd is a respiratory quinol: O₂ oxidoreductase found in many prokaryotes, including a number of pathogens. The main bioenergetic function of the enzyme is the production of a proton motive force by the vectorial charge transfer of protons. The sequences of cytochromes bd are not homologous to those of the other respiratory oxygen reductases, i.e., the heme-copper oxygen reductases or alternative oxidases (AOX). Generally, cytochromes bd are noteworthy for their high affinity for O₂ and resistance to inhibition by cyanide. In E. coli, for example, cytochrome bd (specifically, cytochrome bd-I) is expressed under O₂-limited conditions. Among the members of the bd-family are the so-called cyanide-insensitive quinol oxidases (CIO) which often have a low content of the eponymous heme d but, instead, have heme b in place of heme d in at least a majority of the enzyme population. However, at this point, no sequence motif has been identified to distinguish cytochrome bd (with a stoichiometric complement of heme d) from an enzyme designated as CIO. Members of the bd-family can be subdivided into those which contain either a long or a short hydrophilic connection between transmembrane helices 6 and 7 in subunit I, designated as the Q-loop. However, it is not clear whether there is a functional consequence of this difference. This review summarizes current knowledge on the physiological functions, genetics, structural and catalytic properties of cytochromes bd. Included in this review are descriptions of the intermediates of the catalytic cycle, the proposed site for the reduction of O₂, evidence for a proton channel connecting this active site to the bacterial cytoplasm, and the molecular mechanism by which a membrane potential is generated.

  20. Nitrate Reductase-Deficient Mutants in Barley 1

    PubMed Central

    Somers, David A.; Kuo, Tsung-Min; Kleinhofs, Andris; Warner, Robert L.

    1983-01-01

    Nitrate reductase-deficient barley (Hordeum vulgare L.) mutants were assayed for the presence of a functional molybdenum cofactor determined from the activity of the molybdoenzyme, xanthine dehydrogenase, and for nitrate reductase-associated activities. Rocket immunoelectrophoresis was used to detect nitrate reductase cross-reacting material in the mutants. The cross-reacting material levels of the mutants ranged from 8 to 136% of the wild type and were correlated with their nitrate reductase-associated activities, except for nar 1c, which lacked all associated nitrate reductase activities but had 38% of the wild-type cross-reacting material. The cross-reacting material of two nar 1 mutants, as well as nar 2a, Xno 18, Xno 19, and Xno 29, exhibited rocket immunoprecipitates that were similar to the wild-type enzyme indicating structural homology between the mutant and wild-type nitrate reductase proteins. The cross-reacting materials of the seven remaining nar 1 alleles formed rockets only in the presence of purified wild-type nitrate reductase, suggesting structural modifications of the mutant cross-reacting materials. All nar 1 alleles and Xno 29 had xanthine dehydrogenase activity indicating the presence of functional molybdenum cofactors. These results suggest that nar 1 is the structural gene for nitrate reductase. Mutants nar 2a, Xno 18, and Xno 19 lacked xanthine dehydrogenase activity and are considered to be molybdenum cofactor deficient mutants. Cross-reacting material was not detected in uninduced wild-type or mutant extracts, suggesting that nitrate reductase is synthesized de novo in response to nitrate. Images Fig. 1 Fig. 3 PMID:16662774

  1. The 40Ar/39Ar dating technique applied to planetary sciences

    NASA Astrophysics Data System (ADS)

    Jourdan, F.

    2012-12-01

    The 40Ar/39Ar technique is a powerful geochronological method that can help to unravel the evolution of the solar system. The 40Ar/39Ar system can not only record the timing of volcanic and metamorphic processes on asteroids and planets, it finds domain of predilection in dating impact events throughout the solar system. However, the 40Ar/39Ar method is a robust analytical technique if, and only if, the events to be dated are well understood and data are not over interpreted. Yet, too many 'ages' reported in the literature are still based on over-interpretation of perturbed age spectra which tends to blur the big picture. This presentation is centred on the most recent applications of the 40Ar/39Ar technique applied to planetary material and through several examples, will attempt to demonstrate the benefit of focusing on statistically robust data. For example, 40Ar/39Ar dating of volcanic events on the Moon suggests that volcanism was mostly concentrated between ca. 3.8 and 3.1 Ga but statistical filtering of the data allow identifying a few well-defined eruptive events. The study of lunar volcanism would also benefit from dating of volcanic spherules. Rigorous filtering of the 40Ar/39Ar age database of lunar melt breccias yielded concordant and ages with high precision for two major basins (i.e. Imbrium & Serenitatis) of the Moon. 40Ar/39Ar dating of lunar impact spherules recovered from four different sites and with high- and low-K compositions shows an increase of ages younger than 400 Ma suggesting a recent increase in the impact flux. The impact history of the LL parent body (bodies?) has yet to be well constrained but may mimic the LHB observed on the Moon, which would indicate that the LL parent body was quite large. 40Ar/39Ar dating (in progress) of grains from the asteroid Itokawa recovered by the japanese Hayabusa mission have the potential to constrain the formation history and exposure age of Itokawa and will allow us to compare the results with the

  2. Ar-40/Ar-39 Studies of Martian Meteorite RBT 04262 and Terrestrial Standards

    NASA Technical Reports Server (NTRS)

    Park, J.; Herzog, G. F.; Turrin, B.; Lindsay, F. N.; Delaney, J. S.; Swisher, C. C., III; Nagao, K.; Nyquist, L. E.

    2014-01-01

    Park et al. recently presented an Ar-40/Ar-39 dating study of maskelynite separated from the Martian meteorite RBT 04262. Here we report an additional study of Ar-40/Ar-39 patterns for smaller samples, each consisting of only a few maskelynite grains. Considered as a material for Ar-40/Ar-39 dating, the shock-produced glass maskelynite has both an important strength (relatively high K concentration compared to other mineral phases) and some potentially problematic weaknesses. At Rutgers, we have been analyzing small grains consisting of a single phase to explore local effects that might be averaged and remain hidden in larger samples. Thus, to assess the homogeneity of the RBT maskelynite and for comparison with the results of, we analyzed six approx. 30 microgram samples of the same maskelynite separate they studied. Furthermore, because most Ar-40/Ar-39 are calculated relative to the age of a standard, we present new Ar-40/Ar-39 age data for six standards. Among the most widely used standards are sanidine from Fish Canyon (FCs) and various hornblendes (hb3gr, MMhb-1, NL- 25), which are taken as primary standards because their ages have been determined by independent, direct measurements of K and A-40.

  3. Instrumentation development for planetary in situ 40Ar/39Ar geochronology

    NASA Astrophysics Data System (ADS)

    Davidheiser-Kroll, B.; Morgan, L. E.; Munk, M.; Warner, N. H.; Gupta, S.; Slaybaugh, R.; Harkness, P.; Mark, D. F.

    2015-12-01

    The chronology of the Solar System, particularly the timing of formation of extraterrestrial bodies and their features, is a major outstanding problem in planetary science. Although various chronological methods for in situ geochronology have been proposed (e.g. Rb-Sr, K-Ar), and even applied (K-Ar, Farley et al., 2014), the reliability, accuracy, and applicability of the 40Ar/39Ar method makes it by far the most desirable chronometer for dating extraterrestrial bodies. The method however relies on the neutron irradiation of samples, and thus a neutron source. We will discuss the challenges and feasibility of deploying a passive neutron source to planetary surfaces for the in situ application of the 40Ar/39Ar chronometer. Requirements in generating and shielding neutrons, as well as analyzing samples are discussed, along with an exploration of limitations such as mass, power, and cost. Two potential solutions for the in situ extraterrestrial deployment of the 40Ar/39Ar method will be presented. Although this represents a challenging task, developing the technology to apply the 40Ar/39Ar method on planetary surfaces would represent a major advance towards constraining the timescale of solar system formation and evolution.

  4. Improvements Needed in the 40Ar/39Ar Study of Geomagnetic Excursion Chronology

    NASA Astrophysics Data System (ADS)

    Champion, D. E.; Turrin, B. D.

    2015-12-01

    Our knowledge of the existence and frequency of brief geomagnetic polarity. excursions only increases with time. Precise and accurate 40Ar/39Ar ages will be required to document this, because 25 or more excursions may have occurred within the Brunhes Epoch (780ky) separated in time by as little as 10ky. Excursions are and will dominantly be discovered in mafic, low K2O rocks. Improvements in the analytical protocol to 40Ar/39Ar date low K2O, "young", and thus low 40Arrad rocks are required. While conventional K/Ar dating "worked", the assumption of perfect atmospheric equilibration is flawed. In particular, using a measured isochron intercept (±2s) to embrace an atmospheric intercept assumption turns a 40Ar/39Ar diffusive extraction into a series of "K/Ar-lite" experiments. The near ubiquitous excess 40Ar exhibited in final steps of "matrix" or "groundmass" fractions from whole-rock experiments (no glass, crystals) suggests equilibration with the atmosphere is not achieved. Removing magnetic sample splits (glass?) thought subject to poor argon retention, and crystals subject to 40Ar inheritance are routinely done without documenting different isochrons. Short 15 to 20 minute irradiation times effectively eliminate recoil and dramatically minimize isotopic corrections, and the assumption of equivalence in Ar isotope recoil behavior. Assuming no pressure dependency and constancy of mass discrimination value ignores knowledge from other gas mass spectroscopy (O, H, He, Ne). Dynamic mass spectroscopy in stable isotopic analysis allows routine per mil and 0.1 per mil ratios to be measured. Maintaining more than daily bracketing air pipette measurements at differing pressures, and controlling the range of pressures from each diffusive step will approximate this dynamic precision. Experiments will be discussed that exhibit aspects of 40Ar/39Ar dating protocols with which precision and accuracy can be improved.

  5. Estimate of the 42Ar content in the Earth's atmosphere

    NASA Astrophysics Data System (ADS)

    Barabash, A. S.; Kornoukhov, V. N.; Jants, V. E.

    1997-02-01

    42Ar is a potential source of background in large volume argon-based detectors. The production of the 42Ar isotope both by cosmic rays and by neutrons produced by testing of nuclear weapons is discussed. We demonstrate that main channel of the 42Ar production is from atmospheric testing of nuclear bombs from 1945 to 1962 and the 42Ar content must be less than 1.3 × 10 -23 parts of 42Ar per part of natAr.

  6. Solubilization and Resolution of the Membrane-Bound Nitrite Reductase from Paracoccus Halodenitrificans into Nitrite and Nitric Oxide Reductases

    NASA Technical Reports Server (NTRS)

    Grant, Michael A.; Cronin, Sonja E.; Hochstein, Lawrence I.

    1984-01-01

    Membranes prepared from Paracoccus halodenitrificans reduced nitrite or nitric oxide to nitrous oxide. Extraction of these membranes with the detergent CHAPSO [3-(3-Chlolamidoporopyldimethylammonio)-1-(2- hydroxy-1-propanesulfonate)], followed by ammonium sulfate fractionation of the solubilized proteins, resulted in the separation of nitrite and nitric oxide reductase activities. The fraction containing nitrite reductase activity spectrally resembled a cd-type cytochrome. Several cytochromes were detected in the nitric oxide reductase fraction. Which, if any, of these cytochromes is associated with the reduction of nitric oxide is not clear at this time.

  7. LASER MICROPROBE **4**0Ar/**3**9Ar DATING OF MINERAL GRAINS IN SITU.

    USGS Publications Warehouse

    Sutter, J.F.; Hartung, J.B.

    1984-01-01

    A laser-microprobe attached to a mass spectrometer for **4**0Ar/**3**9Ar age determination of single mineral grains in geological materials has been made operational at the US Geological Survey, Reston, VA. This microanalytical technique involves focusing a pulsed laser beam onto a sample contained in an ultra-high vacuum chamber attached to a rare-gas mass spectrometer. Argon in the neutron-irradiated sample is released by heating with the laser pulse and its isotopic composition is measured to yield an **4**0Ar/**3**9Ar age. Laser probe **4**0Ar/**3**9Ar ages of single mineral grains measured in situ can aid greatly in understanding the chronology of many geological situations where datable minerals are present but are not physically separable in quantities needed for conventional age dating.

  8. Ar-39-Ar-40 Ages of Euerites and the Thermal History of Asteroid 4-Vesta

    NASA Technical Reports Server (NTRS)

    Bogard, Donald D.; Garrison, Daniel H.

    2002-01-01

    Eucrite meteorites are igneous rocks that derive from a large asteroid, probably 4 Vesta. Prior studies have shown that after eucrites formed, most were subsequently metamorphosed to temperatures up to equal to or greater than 800 C, and much later many were brecciated and heated by large impacts into the parent body surface. The uncommon basaltic, unbrecciated eucrites also formed near the surface but presumably escaped later brecciation, whereas the cumulate eucrites formed at depth where metamorphism may have persisted for a considerable period. To further understand the complex HED parent body thermal history, we determined new Ar-39-Ar-40 ages for nine eucrites classified as basaltic but unbrecciated, six eucrites classified as cumulate, and several basaltic-brecciated eucrites. Relatively precise Ar-Ar ages of two cumulate eucrites (Moama and EET87520) and four unbrecciated eucrites give a tight cluster at 4.48 +/1 0.01 Gyr. Ar-Ar ages of six additional unbrecciated eucrites are consistent with this age, within their larger age uncertainties. In contrast, available literature data on Pb-Pb isochron ages of four cumulate eucrites and one unbrecciated eucrite vary over 4.4-4.515 Gyr, and Sm-147 - Nd-143 isochron ages of four cumulate and three unbrecciated eucrites vary over 4.41-4.55 Gyr. Similar Ar-Ar ages for cumulate and unbrecciated eucrites imply that cumulate eucrites do not have a younger formation age than basaltic eucrites, as previously proposed. Rather, we suggest that these cumulate and unbrecciated eucrites resided at depth where parent body temperatures were sufficiently high to cause the K-Ar and some other chronometers to remain open diffusion systems. From the strong clustering of Ar-Ar ages at approximately 4.48 Gyr, we propose that these meteorites were excavated from depth in a single large impact event approximately 4.48 Gyr ago, which quickly cooled the samples and started the K-Ar chronometer. A large (approximately 460 km) crater

  9. Substrate induction of nitrate reductase in barley aleurone layers.

    PubMed

    Ferrari, T E; Varner, J E

    1969-01-01

    Nitrate induces the formation of nitrate reductase activity in barley (Hordeum vulgare L. cv. Himalaya) aleurone layers. Previous work has demonstrated de novo synthesis of alpha-amylase by gibberellic acid in the same tissue. The increase in nitrate reductase activity is inhibited by cycloheximide and 6-methylpurine, but not by actinomycin D. Nitrate does not induce alpha-amylase synthesis, and it has no effect on the gibberellic acid-induced synthesis of alpha-amylase. Also, there is little or no direct effect of gibberellic acid (during the first 6 hr of induction) or of abscisic acid on the nitrate-induced formation of nitrate reductase. Gibberellic acid does interfere with nitrate reductase activity during long-term experiments (greater than 6 hr). However, the time course of this inhibition suggests that the inhibition may be a secondary one. Barley aleurone layers therefore provide a convenient tissue for the study of both substrate- and hormone-induced enzyme formation.

  10. The ArsD As(III) metallochaperone

    PubMed Central

    Ajees, A. Abdul; Yang, Jianbo

    2013-01-01

    Arsenic, a toxic metalloid widely existing in the environment, causes a variety of health problems. The ars operon encoded by Escherichia coli plasmid R773 has arsD and arsA genes, where ArsA is an ATPase that is the catalytic subunit of the ArsAB As(III) extrusion pump, and ArsD is an arsenic chaperone for ArsA. ArsD transfers As(III) to ArsA and increases the affinity of ArsA for As(III), allowing resistance to environmental concentrations of arsenic. Cys12, Cys13 and Cys18 in ArsD form a three sulfur-coordinated As(III) binding site that is essential for metallochaperone activity. ATP hydrolysis by ArsA is required for transfer of As(III) from ArsD to ArsA, suggesting that transfer occurs with a conformation of ArsA that transiently forms during the catalytic cycle. The 1.4 Å x-ray crystal structure of ArsD shows a core of four β-strands flanked by four α-helices in a thioredoxin fold. Docking of ArsD with ArsA was modeled in silico. Independently ArsD mutants exhibiting either weaker or stronger interaction with ArsA were selected. The locations of the mutations mapped on the surface of ArsD are consistent with the docking model. The results suggest that the interface with ArsA involves one surface of α1 helix and metalloid binding site of ArsD. PMID:21188475

  11. Ar-39-Ar-40 Evidence for Early Impact Events on the LL Parent Body

    NASA Technical Reports Server (NTRS)

    Dixon, E. T.; Bogard, D. D.; Garrison, D. H.; Rubin, A. E.

    2006-01-01

    We determined Ar-39-Ar-40 ages of eight LL chondrites, and one igneous inclusion from an LL chondrite, with the object of understanding the thermal history of the LL-chondrite parent body. The meteorites in this study have a range of petrographic types from LL3.3 to LL6, and shock stages from S1 to S4. These meteorites reveal a range of K-Ar ages from 23.66 to 24.50 Ga, and peak ages from 23.74 to 24.55 Ga. Significantly, three of the eight chondrites (LL4, 5, 6) have K-Ar ages of -4.27 Ga. One of these (MIL99301) preserves an Ar-39-Ar-40 age of 4.23 +/- 0.03 Ga from low-temperature extractions, and an older age of 4.52 +/- 0.08 Ga from the highest temperature extractions. In addition, an igneous-textured impact melt DOM85505,22 has a peak Ar-39-Ar-40 age of >= 4.27 Ga. We interpret these results as evidence for impact events that occurred at about 4.27 Ga on the LL parent body that produced local impact melts, reset the Ar-39-Ar-40 ages of some meteorites, and exhumed (or interred) others, resulting in a range of cooling ages. The somewhat younger peak age of 3.74 Ga from GR095658 (LL3.3) suggests an additional impact event close to timing of impact-reset ages of some other ordinary chondrites between 3.6-3.8 Ga. The results from MIL99301 suggest that some apparently unshocked (Sl) chondrites may have substantially reset Ar-39-Ar-40 ages. A previous petrographic investigation of MIL99301 suggested that reheating to temperatures less than or equal to type 4 petrographic conditions (600C) caused fractures in olivine to anneal, resulting in a low apparent shock stage of S1 (unshocked). The Ar-39-Ar-40 age spectrum of MIL99301 is consistent with this interpretation. Older ages from high-T extractions may date an earlier impact event at 4.52 +/- 0.08 Ga, whereas younger ages from lower-T extractions date a later impact event at 4.23 Ar-39-Ar-40 0.03 Ga that may have caused annealing of feldspar and olivine

  12. 40Ar/39Ar age of Cretaceous-Tertiary boundary tektites from Haiti

    USGS Publications Warehouse

    Izett, G.A.; Dalrymple, G.B.; Snee, L.W.

    1991-01-01

    40Ar/39Ar dating of tektites discovered recently in Cretaceous-Tertiary (K-T) boundary marine sedimentary rocks on Haiti indicates that the K-T boundary and impact event are coeval at 64.5 ?? 0.1 million years ago. Sanidine from a bentonite that lies directly above the K-T boundary in continental, coal-bearing, sedimentary rocks of Montana was also dated and has a 40Ar/39Ar age of 64.6 ?? 0.2 million years ago, which is indistinguishable statistically from the age of the tektites.

  13. Rapid kimberlite ascent and the significance of Ar-Ar ages in xenolith phlogopites

    PubMed

    Kelley; Wartho

    2000-07-28

    Kimberlite eruptions bring exotic rock fragments and minerals, including diamonds, from deep within the mantle up to the surface. Such fragments are rapidly absorbed into the kimberlite magma so their appearance at the surface implies rapid transport from depth. High spatial resolution Ar-Ar age data on phlogopite grains in xenoliths from Malaita in the Solomon Islands, southwest Pacific, and Elovy Island in the Kola Peninsula, Russia, indicate transport times of hours to days depending upon the magma temperature. In addition, the data show that the phlogopite grains preserve Ar-Ar ages recorded at high temperature in the mantle, 700 degrees C above the conventional closure temperature.

  14. 40Ar/39Ar ages of the AD 79 eruption of Vesuvius, Italy

    USGS Publications Warehouse

    Lanphere, M.; Champion, D.; Melluso, L.; Morra, V.; Perrotta, A.; Scarpati, C.; Tedesco, D.; Calvert, A.

    2007-01-01

    The Italian volcano, Vesuvius, erupted explosively in AD 79. Sanidine from pumice collected at Casti Amanti in Pompeii and Villa Poppea in Oplontis yielded a weighted-mean 40Ar/39Ar age of 1925??66 years in 2004 (1?? uncertainty) from incremental-heating experiments of eight aliquants of sanidine. This is the calendar age of the eruption. Our results together with the work of Renne et al. (1997) and Renne and Min (1998) demonstrate the validity of the 40Ar/39Ar method to reconstruct the recent eruptive history of young, active volcanoes. ?? Springer-Verlag 2006.

  15. 40Ar/39Ar technique of KAr dating: a comparison with the conventional technique

    USGS Publications Warehouse

    Brent, Dalrymple G.; Lanphere, M.A.

    1971-01-01

    K-Ar ages have been determined by the 40Ar/39Ar total fusion technique on 19 terrestrial samples whose conventional K-Ar ages range from 3.4 my to nearly 1700 my. Sample materials included biotite, muscovite, sanidine, adularia, plagioclase, hornblende, actinolite, alunite, dacite, and basalt. For 18 samples there are no significant differences at the 95% confidence level between the KAr ages obtained by these two techniques; for one sample the difference is 4.3% and is statistically significant. For the neutron doses used in these experiments (???4 ?? 1018 nvt) it appears that corrections for interfering Ca- and K-derived Ar isotopes can be made without significant loss of precision for samples with K/Ca > 1 as young as about 5 ?? 105 yr, and for samples with K/Ca < 1 as young as about 107 yr. For younger samples the combination of large atmospheric Ar corrections and large corrections for Ca- and K-derived Ar may make the precision of the 40Ar/39Ar technique less than that of the conventional technique unless the irradiation parameters are adjusted to minimize these corrections. ?? 1971.

  16. Rapid kimberlite ascent and the significance of Ar-Ar ages in xenolith phlogopites

    PubMed

    Kelley; Wartho

    2000-07-28

    Kimberlite eruptions bring exotic rock fragments and minerals, including diamonds, from deep within the mantle up to the surface. Such fragments are rapidly absorbed into the kimberlite magma so their appearance at the surface implies rapid transport from depth. High spatial resolution Ar-Ar age data on phlogopite grains in xenoliths from Malaita in the Solomon Islands, southwest Pacific, and Elovy Island in the Kola Peninsula, Russia, indicate transport times of hours to days depending upon the magma temperature. In addition, the data show that the phlogopite grains preserve Ar-Ar ages recorded at high temperature in the mantle, 700 degrees C above the conventional closure temperature. PMID:10915621

  17. 40Ar/39Ar Age of Cretaceous-Tertiary Boundary Tektites from Haiti.

    PubMed

    Izett, G A; Dalrymple, G B; Snee, L W

    1991-06-14

    (40)Ar/(39)Ar dating of tektites discovered recently in Cretaceous-Tertiary (K-T) boundary marine sedimentary rocks on Haiti indicates that the K-T boundary and impact event are coeval at 64.5 +/- 0.1 million years ago. Sanidine from a bentonite that lies directly above the K-T boundary in continental, coal-bearing, sedimentary rocks of Montana was also dated and has a (40)Ar/(39)Ar age of 64.6 +/- 0.2 million years ago, which is indistinguishable statistically from the age of the tektites.

  18. 40Ar/39Ar ages in deformed potassium feldspar: evidence of microstructural control on Ar isotope systematics

    NASA Astrophysics Data System (ADS)

    Reddy, Steven M.; Potts, Graham J.; Kelley, Simon P.

    2001-05-01

    Detailed field and microstructural studies have been combined with high spatial resolution ultraviolet laser 40Ar/39Ar dating of naturally deformed K-feldspar to investigate the direct relationship between deformation-related microstructure and Ar isotope systematics. The sample studied is a ~1,000 Ma Torridonian arkose from Skye, Scotland, that contains detrital feldspars previously metamorphosed at amphibolite-facies conditions ~1,700 Ma. The sample was subsequently deformed ~430 Ma ago during Caledonian orogenesis. The form and distribution of deformation-induced microstructures within three different feldspar clasts has been mapped using atomic number contrast and orientation contrast imaging, at a range of scales, to identify intragrain variations in composition and lattice orientation. These variations have been related to thin section and regional structural data to provide a well-constrained deformation history for the feldspar clasts. One hundred and forty-three in-situ 40Ar/39Ar analyses measured using ultraviolet laser ablation record a range of apparent ages (317-1030 Ma). The K-feldspar showing the least strain records the greatest range of apparent ages from 420-1,030 Ma, with the oldest apparent ages being found close to the centre of the feldspar away from fractures and the detrital grain boundary. The most deformed K-feldspar yields the youngest apparent ages (317-453 Ma) but there is no spatial relationship between apparent age and the detrital grain boundary. Within this feldspar, the oldest apparent ages are recorded from orientation domain boundaries and fracture surfaces where an excess or trapped 40Ar component resides. Orientation contrast images at a similar scale to the Ar analyses illustrate a significant deformation-related microstructural difference between the feldspars and we conclude that deformation plays a significant role in controlling Ar systematics of feldspars at both the inter- and intragrain scales even at relatively low

  19. Regulation of the Neurospora crassa assimilatory nitrate reductase.

    PubMed Central

    Ketchum, P A; Zeeb, D D; Owens, M S

    1977-01-01

    Reduced nicotinamide adenine dinucleotide phosphate (NADPH)-nitrate reductase from Neurospora crassa was purified and found to be stimulated by certain amino acids, citrate, and ethylenediaminetetraacetic acid (EDTA). Stimulation by citrate and the amino acids was dependent upon the prior removal of EDTA from the enzyme preparations, since low quantities of EDTA resulted in maximal stimulation. Removal of EDTA from enzyme preparations by dialysis against Chelex-containing buffer resulted in a loss of nitrate reductase activity. Addition of alanine, arginine, glycine, glutamine, glutamate, histidine, tryptophan, and citrate restored and stimulated nitrate reductase activity from 29- to 46-fold. The amino acids tested altered the Km of NADPH-nitrate reductase for NADPH but did not significantly change that for nitrate. The Km of nitrate reductase for NADPH increased with increasing concentrations of histidine but decreased with increasing concentrations of glutamine. Amino acid modulation of NADPH-nitrate reductase activity is discussed in relation to the conservation of energy (NADPH) by Neurospora when nitrate is the nitrogen source. PMID:19423

  20. An overview on 5alpha-reductase inhibitors.

    PubMed

    Aggarwal, Saurabh; Thareja, Suresh; Verma, Abhilasha; Bhardwaj, Tilak Raj; Kumar, Manoj

    2010-02-01

    Benign prostatic hyperplasia (BPH) is the noncancerous proliferation of the prostate gland associated with benign prostatic obstruction and lower urinary tract symptoms (LUTS) such as frequency, hesitancy, urgency, etc. Its prevalence increases with age affecting around 70% by the age of 70 years. High activity of 5alpha-reductase enzyme in humans results in excessive dihydrotestosterone levels in peripheral tissues and hence suppression of androgen action by 5alpha-reductase inhibitors is a logical treatment for BPH as they inhibit the conversion of testosterone to dihydrotestosterone. Finasteride (13) was the first steroidal 5alpha-reductase inhibitor approved by U.S. Food and Drug Administration (USFDA). In human it decreases the prostatic DHT level by 70-90% and reduces the prostatic size. Dutasteride (27) another related analogue has been approved in 2002. Unlike Finasteride, Dutasteride is a competitive inhibitor of both 5alpha-reductase type I and type II isozymes, reduced DHT levels >90% following 1 year of oral administration. A number of classes of non-steroidal inhibitors of 5alpha-reductase have also been synthesized generally by removing one or more rings from the azasteroidal structure or by an early non-steroidal lead (ONO-3805) (261). In this review all categories of inhibitors of 5alpha-reductase have been covered. PMID:19879888

  1. Effects of thioredoxin reductase-1 deletion on embryogenesis and transcriptome

    PubMed Central

    Bondareva, Alla A.; Capecchi, Mario R.; Iverson, Sonya V.; Li, Yan; Lopez, Nathan I.; Lucas, Olivier; Merrill, Gary F.; Prigge, Justin R.; Siders, Ashley M.; Wakamiya, Maki; Wallin, Stephanie L.; Schmidt, Edward E.

    2007-01-01

    Thioredoxin reductases (Txnrd)1 maintain intracellular redox homeostasis in most organisms. Metazoans Txnrds also participate in signal transduction. Mouse embryos homozygous for a targeted null mutation of the txnrd1 gene, encoding the cytosolic thioredoxin reductase, were viable at embryonic day 8.5 (E8.5) but not at E9.5. Histology revealed that txnrd1−/− cells were capable of proliferation and differentiation; however, mutant embryos were smaller than wild-type littermates and failed to gastrulate. In situ marker gene analyses indicated primitive streak mesoderm did not form. Microarray analyses on E7.5 txnrd−/− and txnrd+/+ littermates showed similar mRNA levels for peroxiredoxins, glutathione reductases, mitochondrial Txnrd2, and most markers of cell proliferation. Conversely, mRNAs encoding sulfiredoxin, IGF-binding protein 1, carbonyl reductase 3, glutamate cysteine ligase, glutathione S-transferases, and metallothioneins were more abundant in mutants. Many gene expression responses mirrored those in thioredoxin reductase 1-null yeast; however mice exhibited a novel response within the peroxiredoxin catalytic cycle. Thus, whereas yeast induce peroxiredoxin mRNAs in response to thioredoxin reductase disruption, mice induced sulfiredoxin mRNA. In summary, Txnrd1 was required for correct patterning of the early embryo and progression to later development. Conserved responses to Txnrd1 disruption likely allowed proliferation and limited differentiation of the mutant embryo cells. PMID:17697936

  2. Opposing Effects of Cyclooxygenase-2 (COX-2) on Estrogen Receptor β (ERβ) Response to 5α-Reductase Inhibition in Prostate Epithelial Cells.

    PubMed

    Liu, Teresa T; Grubisha, Melanie J; Frahm, Krystle A; Wendell, Stacy G; Liu, Jiayan; Ricke, William A; Auchus, Richard J; DeFranco, Donald B

    2016-07-01

    Current pharmacotherapies for symptomatic benign prostatic hyperplasia (BPH), an androgen receptor-driven, inflammatory disorder affecting elderly men, include 5α-reductase (5AR) inhibitors (i.e. dutasteride and finasteride) to block the conversion of testosterone to the more potent androgen receptor ligand dihydrotestosterone. Because dihydrotestosterone is the precursor for estrogen receptor β (ERβ) ligands, 5AR inhibitors could potentially limit ERβ activation, which maintains prostate tissue homeostasis. We have uncovered signaling pathways in BPH-derived prostate epithelial cells (BPH-1) that are impacted by 5AR inhibition. The induction of apoptosis and repression of the cell adhesion protein E-cadherin by the 5AR inhibitor dutasteride requires both ERβ and TGFβ. Dutasteride also induces cyclooxygenase type 2 (COX-2), which functions in a negative feedback loop in TGFβ and ERβ signaling pathways as evidenced by the potentiation of apoptosis induced by dutasteride or finasteride upon pharmacological inhibition or shRNA-mediated ablation of COX-2. Concurrently, COX-2 positively impacts ERβ action through its effect on the expression of a number of steroidogenic enzymes in the ERβ ligand metabolic pathway. Therefore, effective combination pharmacotherapies, which have included non-steroidal anti-inflammatory drugs, must take into account biochemical pathways affected by 5AR inhibition and opposing effects of COX-2 on the tissue-protective action of ERβ. PMID:27226548

  3. A comparison of glucose oxidase and aldose dehydrogenase as mediated anodes in printed glucose/oxygen enzymatic fuel cells using ABTS/laccase cathodes.

    PubMed

    Jenkins, Peter; Tuurala, Saara; Vaari, Anu; Valkiainen, Matti; Smolander, Maria; Leech, Dónal

    2012-10-01

    Current generation by mediated enzyme electron transfer at electrode surfaces can be harnessed to provide biosensors and redox reactions in enzymatic fuel cells. A glucose/oxygen enzymatic fuel cell can provide power for portable and implantable electronic devices. High volume production of enzymatic fuel cell prototypes will likely require printing of electrode and catalytic materials. Here we report on preparation and performance of, completely enzymatic, printed glucose/oxygen biofuel cells. The cells are based on filter paper coated with conducting carbon inks, enzyme and mediator. A comparison of cell performance using a range of mediators for either glucose oxidase (GOx) or aldose dehydrogenase (ALDH) oxidation of glucose at the anode and ABTS and a fungal laccase, for reduction of oxygen at the cathode, is reported. Highest power output, although of limited stability, is observed for ALDH anodes mediated by an osmium complex, providing a maximum power density of 3.5 μW cm(-2) at 0.34 V, when coupled to a laccase/ABTS cathode. The stability of cell voltage in a biobattery format, above a threshold of 200 mV under a moderate 75 kΩ load, is used to benchmark printed fuel cell performance. Highest stability is obtained for printed fuel cells using ALDH, providing cell voltages over the threshold for up to 74 h, compared to only 2 h for cells with anodes using GOx. These results provide promising directions for further development of mass-producible, completely enzymatic, printed biofuel cells.

  4. A comparison of glucose oxidase and aldose dehydrogenase as mediated anodes in printed glucose/oxygen enzymatic fuel cells using ABTS/laccase cathodes.

    PubMed

    Jenkins, Peter; Tuurala, Saara; Vaari, Anu; Valkiainen, Matti; Smolander, Maria; Leech, Dónal

    2012-10-01

    Current generation by mediated enzyme electron transfer at electrode surfaces can be harnessed to provide biosensors and redox reactions in enzymatic fuel cells. A glucose/oxygen enzymatic fuel cell can provide power for portable and implantable electronic devices. High volume production of enzymatic fuel cell prototypes will likely require printing of electrode and catalytic materials. Here we report on preparation and performance of, completely enzymatic, printed glucose/oxygen biofuel cells. The cells are based on filter paper coated with conducting carbon inks, enzyme and mediator. A comparison of cell performance using a range of mediators for either glucose oxidase (GOx) or aldose dehydrogenase (ALDH) oxidation of glucose at the anode and ABTS and a fungal laccase, for reduction of oxygen at the cathode, is reported. Highest power output, although of limited stability, is observed for ALDH anodes mediated by an osmium complex, providing a maximum power density of 3.5 μW cm(-2) at 0.34 V, when coupled to a laccase/ABTS cathode. The stability of cell voltage in a biobattery format, above a threshold of 200 mV under a moderate 75 kΩ load, is used to benchmark printed fuel cell performance. Highest stability is obtained for printed fuel cells using ALDH, providing cell voltages over the threshold for up to 74 h, compared to only 2 h for cells with anodes using GOx. These results provide promising directions for further development of mass-producible, completely enzymatic, printed biofuel cells. PMID:22200380

  5. 40Ar/39Ar systematics and argon diffusion in amber: implications for ancient earth atmospheres

    USGS Publications Warehouse

    Landis, G.P.; Snee, L.W.

    1991-01-01

    Argon isotope data indicate retained argon in bulk amber (matrix gas) is radiogenic [40Ar/39Ar ???32o] than the much more abundant surface absorbed argon [40Ar/39Ar ???295.5]. Neutron-induced 39Ar is retained in amber during heating experiments to 150?? -250??C, with no evidence of recoiled 39Ar found after irradiation. A maximum permissible volume diffusion coefficient of argon in amber (at ambient temperature) D???1.5 x 10-17 cm2S-1 is calculated from 39Ar retention. 40Ar/39Ar age calculations indicate Dominican Republic amber is ??? 45 Ma and North Dakota amber is ??? 89 Ma, both at least reasonable ages for the amber based upon stratigraphic and paleontological constraints and upon the small amount of radiogenic 40Ar. To date, over 300 gas analyses of ambers and resins of Cretaceous to Recent age that are geographically distributed among fifteen noted world locations identify mixtures of gases in different sites within amber (Berner and Landis, 1988). The presence of multiple mixing trends between compositionally distinct end-members gases within the same sample and evidence for retained radiogenic argon within the amber argue persuasivley against rapid exchange by diffusion of amber-contained gases with moder air. Only gas in primary bubbles entrapped between successive flows of tree resin has been interpreted as original "ancient air", which is an O2-rich end-member gas with air-like N2/Ar ratios. Gas analyses of these primary bubbles indicate atmospheric O2 levels in the Late Cretaceous of ??? 35%, and that atmospheric O2 dropped by early Tertiary time to near a present atmospheric level of 21% O2. A very low argon diffusion coefficient in amber persuasively argues for a gas in primary bubbles trapped in amber being ancient air (possibly modified only by O2 reaction with amber). ?? 1991.

  6. Role of 5 alpha-reductase in health and disease.

    PubMed

    Randall, V A

    1994-04-01

    The mechanism of androgen action varies in different tissues, but in the majority of androgen target tissues either testosterone or 5 alpha-dihydrotestosterone (DHT) binds to a specific androgen receptor to form a complex that can regulate gene expression. Testosterone is metabolized to DHT by the enzyme 5 alpha-reductase. The autosomal recessive genetic disorder of 5 alpha-reductase deficiency has clearly shown that the requirement for DHT formation varies with different tissues. In this syndrome genetic males contain normal male internal structures including testes, but exhibit ambiguous or female external genitalia at birth; at puberty they undergo partial virilization which includes development of a male gender identity even if brought up as females. Their development suggests that testosterone itself is able to stimulate psychosexual behaviour, development of the embryonic wolffian duct, muscle development, voice deepening, spermatogenesis, and axillary and pubic hair growth; DHT seems to be essential for prostate development and growth, the development of the external genitalia and male patterns of facial and body hair growth or male-pattern baldness. How different hormones operate to regulate genes via the same receptor is currently unknown, but appears to involve cell-specific factors. The 5-alpha-reductase enzyme has proved difficult to isolate biochemically, but recently at least two human isoenzymes have been identified using molecular biological methods. All the various 5 alpha-reductase-deficient kindreds have been shown to have mutations in 5 alpha-reductase 2, the predominant form in the prostate. The biological role of 5 alpha-reductase 1 has not yet been ascertained, but at present it cannot be ruled out that some of the actions ascribed to testosterone are indeed in cells producing DHT via this enzyme. The activity of 5 alpha-reductase is also implicated in benign prostatic hypertrophy, hirsutism and possibly male-pattern baldness; recent evidence

  7. ArsP: a methylarsenite efflux permease

    PubMed Central

    Chen, Jian; Madegowda, Mahendra; Bhattacharjee, Hiranmoy; Rosen, Barry P.

    2015-01-01

    Trivalent organoarsenic compounds are far more toxic than either pentavalent organoarsenicals or inorganic arsenite. Many microbes methylate inorganic arsenite (As(III)) to more toxic and carcinogenic methylarsenite (MAs(III)). Additionally, monosodium methylarsenate (MSMA or MAs(V)) has been used widely as an herbicide and is reduced by microbial communities to MAs(III). Roxarsone (3-nitro-4-hydroxybenzenearsonic acid) is a pentavalent aromatic arsenical that is used as antimicrobial growth promoter for poultry and swine, and its active form is the trivalent species Rox(III). A bacterial permease, ArsP, from Campylobacter jejuni, was recently shown to confer resistance to roxarsone. In this study C. jejuni arsP was expressed in Escherichia coli and shown to confer resistance to MAs(III) and Rox(III) but not to inorganic As(III) or pentavalent organoarsenicals. Cells of E. coli expressing arsP did not accumulate trivalent organoarsenicals. Everted membrane vesicles from those cells accumulated MAs(III)>Rox(III) with energy supplied by NADH oxidation, reflecting efflux from cells. The vesicles did not transport As(III), MAs(V) or pentavalent roxarsone. Mutation or modification of the two conserved cysteine residues resulted in loss of transport activity, suggesting that they play a role in ArsP function. Thus ArsP is the first identified efflux system specific for trivalent organoarsenicals. PMID:26234817

  8. Correlation diagrams in 40 Ar/39Ar dating: is there a correct choice?

    USGS Publications Warehouse

    Dalrymple, G.B.; Lanphere, M.A.; Pringle, M.S.

    1988-01-01

    Contrary to published assertions, the 2 types of correlation diagrams used in the interpretation of 40Ar/39Ar incremental-heating data yield the same information provided the correct mathematics are used for estimating correlation coefficients and for the least squares fit. The choice is simply between 2 illustrative, graphical displays, neither of which is fundamentally superior to the other. -Authors

  9. Wolinella succinogenes quinol:fumarate reductase and its comparison to E. coli succinate:quinone reductase.

    PubMed

    Lancaster, C Roy D

    2003-11-27

    The three-dimensional structure of Wolinella succinogenes quinol:fumarate reductase (QFR), a dihaem-containing member of the superfamily of succinate:quinone oxidoreductases (SQOR), has been determined at 2.2 A resolution by X-ray crystallography [Lancaster et al., Nature 402 (1999) 377-385]. The structure and mechanism of W. succinogenes QFR and their relevance to the SQOR superfamily have recently been reviewed [Lancaster, Adv. Protein Chem. 63 (2003) 131-149]. Here, a comparison is presented of W. succinogenes QFR to the recently determined structure of the mono-haem containing succinate:quinone reductase from Escherichia coli [Yankovskaya et al., Science 299 (2003) 700-704]. In spite of differences in polypeptide and haem composition, the overall topology of the membrane anchors and their relative orientation to the conserved hydrophilic subunits is strikingly similar. A major difference is the lack of any evidence for a 'proximal' quinone site, close to the hydrophilic subunits, in W. succinogenes QFR.

  10. Isolation and Characterization of cDNAs Encoding Leucoanthocyanidin Reductase and Anthocyanidin Reductase from Populus trichocarpa

    PubMed Central

    Lu, Wanxiang; Yang, Li; Karim, Abdul; Luo, Keming

    2013-01-01

    Proanthocyanidins (PAs) contribute to poplar defense mechanisms against biotic and abiotic stresses. Transcripts of PA biosynthetic genes accumulated rapidly in response to infection by the fungus Marssonina brunnea f.sp. multigermtubi, treatments of salicylic acid (SA) and wounding, resulting in PA accumulation in poplar leaves. Anthocyanidin reductase (ANR) and leucoanthocyanidin reductase (LAR) are two key enzymes of the PA biosynthesis that produce the main subunits: (+)-catechin and (−)-epicatechin required for formation of PA polymers. In Populus, ANR and LAR are encoded by at least two and three highly related genes, respectively. In this study, we isolated and functionally characterized genes PtrANR1 and PtrLAR1 from P. trichocarpa. Phylogenetic analysis shows that Populus ANR1 and LAR1 occurr in two distinct phylogenetic lineages, but both genes have little difference in their tissue distribution, preferentially expressed in roots. Overexpression of PtrANR1 in poplar resulted in a significant increase in PA levels but no impact on catechin levels. Antisense down-regulation of PtrANR1 showed reduced PA accumulation in transgenic lines, but increased levels of anthocyanin content. Ectopic expression of PtrLAR1 in poplar positively regulated the biosynthesis of PAs, whereas the accumulation of anthocyanin and flavonol was significantly reduced (P<0.05) in all transgenic plants compared to the control plants. These results suggest that both PtrANR1 and PtrLAR1 contribute to PA biosynthesis in Populus. PMID:23741362

  11. Equine 5α-reductase activity and expression in epididymis.

    PubMed

    Corbin, C J; Legacki, E L; Ball, B A; Scoggin, K E; Stanley, S D; Conley, A J

    2016-10-01

    The 5α-reductase enzymes play an important role during male sexual differentiation, and in pregnant females, especially equine species where maintenance relies on 5α-reduced progesterone, 5α-dihydroprogesterone (DHP). Epididymis expresses 5α-reductases but was not studied elaborately in horses. Epididymis from younger and older postpubertal stallions was divided into caput, corpus and cauda and examined for 5α-reductase activity and expression of type 1 and 2 isoforms by quantitative real-time polymerase chain reaction (qPCR). Metabolism of progesterone and testosterone to DHP and dihydrotestosterone (DHT), respectively, by epididymal microsomal protein was examined by thin-layer chromatography and verified by liquid chromatography tandem mass spectrometry (LC-MS/MS). Relative inhibitory potencies of finasteride and dutasteride toward equine 5α-reductase activity were investigated. Pregnenolone was investigated as an additional potential substrate for 5α-reductase, suggested previously from in vivo studies in mares but never directly examined. No regional gradient of 5α-reductase expression was observed by either enzyme activity or transcript analysis. Results of PCR experiments suggested that type 1 isoform predominates in equine epididymis. Primers for the type 2 isoform were unable to amplify product from any samples examined. Progesterone and testosterone were readily reduced to DHP and DHT, and activity was effectively inhibited by both inhibitors. Using epididymis as an enzyme source, no experimental evidence was obtained supporting the notion that pregnenolone could be directly metabolized by equine 5α-reductases as has been suggested by previous investigators speculating on alternative metabolic pathways leading to DHP synthesis in placenta during equine pregnancies. PMID:27466384

  12. Equine 5α-reductase activity and expression in epididymis.

    PubMed

    Corbin, C J; Legacki, E L; Ball, B A; Scoggin, K E; Stanley, S D; Conley, A J

    2016-10-01

    The 5α-reductase enzymes play an important role during male sexual differentiation, and in pregnant females, especially equine species where maintenance relies on 5α-reduced progesterone, 5α-dihydroprogesterone (DHP). Epididymis expresses 5α-reductases but was not studied elaborately in horses. Epididymis from younger and older postpubertal stallions was divided into caput, corpus and cauda and examined for 5α-reductase activity and expression of type 1 and 2 isoforms by quantitative real-time polymerase chain reaction (qPCR). Metabolism of progesterone and testosterone to DHP and dihydrotestosterone (DHT), respectively, by epididymal microsomal protein was examined by thin-layer chromatography and verified by liquid chromatography tandem mass spectrometry (LC-MS/MS). Relative inhibitory potencies of finasteride and dutasteride toward equine 5α-reductase activity were investigated. Pregnenolone was investigated as an additional potential substrate for 5α-reductase, suggested previously from in vivo studies in mares but never directly examined. No regional gradient of 5α-reductase expression was observed by either enzyme activity or transcript analysis. Results of PCR experiments suggested that type 1 isoform predominates in equine epididymis. Primers for the type 2 isoform were unable to amplify product from any samples examined. Progesterone and testosterone were readily reduced to DHP and DHT, and activity was effectively inhibited by both inhibitors. Using epididymis as an enzyme source, no experimental evidence was obtained supporting the notion that pregnenolone could be directly metabolized by equine 5α-reductases as has been suggested by previous investigators speculating on alternative metabolic pathways leading to DHP synthesis in placenta during equine pregnancies.

  13. Comparative 40Ar/39Ar and K-Ar dating of illite-type clay minerals: A tentative explanation for age identities and differences

    NASA Astrophysics Data System (ADS)

    Clauer, Norbert; Zwingmann, Horst; Liewig, Nicole; Wendling, Raymond

    2012-10-01

    The 40K/40Ar (K-Ar) and 40Ar/39Ar dating methods are applied here to the same, very small, micrometric illite-type particles that crystallized under low-temperature (< 175 °C) hydrothermal conditions in deeply buried Rotliegend (Permian) gas-bearing sandstones of NW Germany. Four samples with a total of fifteen size fractions from < 2 to 20-40 μm yield K-Ar ages that range from 166.0 ± 3.4 to 214.0 ± 5.9 Ma. The same size fractions dated by the 40Ar/39Ar method give total-gas ages ranging from 173.3 ± 2.0 to 228.8 ± 1.6 Ma. Nearly all 40Ar/39Ar total-gas ages are slightly older, which cannot be explained by the recoil effect only, the impact of which being amplified by the inhomogeneous shape of the clay minerals and their crystallographic characteristics, with varied crystallinity indices, and a particle width about 10 times large than thickness. The 40Ar/39Ar data outline some advantages, such as the plateaus obtained by incremental step heating of the various size fractions, even if not translatable straight as ages of the illite populations; they allow identification of two generations of authigenic illite that formed at about 200 and 175 Ma, and one detrital generation. However, 40Ar/39Ar dating of clay minerals remains challenging as technical factors, such as the non-standardized encapsulation, may have potential unexpected effects. Both dating methods have their limitations: (1) K-Ar dating requires relatively large samples (ca. 10-20 mg) incurring potential sample homogeneity problems, with two aliquots required for K and Ar analysis for an age determination, also inducing a higher analytical uncertainty; (2) an identified drawback of 40Ar/39Ar dating is Ar recoil and therefore potential loss that occurs during neutronic creation of 39Ar from 39K, mostly in the finer mineral particles. If all the recoiled 39Ar is redistributed into adjacent grains/minerals, the final 40Ar/39Ar age of the analyzed size fraction remains theoretically identical, but it

  14. Age and Obesity Promote Methylation and Suppression of 5-Alpha Reductase 2–Implications for Personalized Therapy in Benign Prostatic Hyperplasia

    PubMed Central

    Bechis, Seth K.; Otsetov, Alexander G.; Ge, Rongbin; Wang, Zongwei; Vangel, Mark G.; Wu, Chin-Lee; Tabatabaei, Shahin; Olumi, Aria F.

    2016-01-01

    Purpose 5α reductase inhibitors (5ARIs) are a main modality of treatment for men suffering from symptomatic benign prostatic hyperplasia (BPH). Over 30% of men do not respond to the therapeutic effects of 5ARIs. We have found that 1/3 of adult prostate samples do not express 5AR2 secondary to epigenetic modifications. We sought to evaluate whether 5AR2 expression in BPH specimens of symptomatic men was linked to methylation of the 5AR2 gene promoter and identify associations with age, obesity, cardiac risk factors, and prostate specific antigen (PSA). Materials and Methods Prostate samples from men undergoing transurethral prostate resection were used. 5AR2 protein expression and gene promoter methylation status were determined by common assays. Clinical variables included age, body mass index (BMI), hypertension, hyperlipidemia, diabetes, PSA, and prostate volume. Univariate and multivariate statistical analyses were performed, followed by stepwise logistic regression modeling. Results BMI and age were significantly correlated with methylation of the 5AR2 gene promoter (p<0.05), whereas prostate volume, PSA, or use of BPH medication were not. Methylation was highly correlated with 5AR protein expression (p<0.0001). In a predictive model, both increasing age and BMI significantly predicted methylation status and protein expression (p<0.01). Conclusions Increasing age and BMI correlate with increased 5AR2 gene promoter methylation and decreased protein expression in men with symptomatic BPH. These results highlight the interplay between age, obesity and gene regulation. Our findings suggest the presence of an individualized epigenetic signature for symptomatic BPH, which may be important for choosing appropriate personalized treatment options. PMID:25916673

  15. Hazard evaluation for 244-AR vault facility

    SciTech Connect

    BRAUN, D.J.

    1999-08-25

    This document presents the results of a hazard identification and evaluation performed on the 244-AR Vault Facility to close a USQ (USQ No.TF-98-0785, Potential Inadequacy in Authorization Basis (PIAB): To Evaluate Miscellaneous Facilities Listed In HNF-2503 And Not Addressed In The TWRS Authorization Basis) that was generated as part of an evaluation of inactive TWRS facilities. A hazard evaluation for the Hanford Site 244-AR Vault Facility was performed. The process and results of the hazard evaluation are provided in this document. A previous hazard evaluation was performed for the 244-AR Vault Facility in 1996 in support of the Basis for Interim Operation (BIO) (HNF-SD-WM-BIO-001, 1998, Revision 1) of the Tank Waste Remediation System (TWRS). The results of that evaluation are provided in the BIO. Upon review of those results it was determined that hazardous conditions that could lead to the release of radiological and toxicological material from the 244-AR vaults due to flooding was not addressed in the original hazards evaluation. This supplemental hazard evaluation addresses this oversight of the original hazard evaluation. The results of the hazard evaluation were compared to the current TWRS BIO to identify any hazardous conditions where Authorization Basis (AB) controls may not be sufficient or may not exist. This document is not part of the AB and is not a vehicle for requesting changes to the AB. It is only intended to provide information about hazardous conditions associated with the condition and configuration of the 244-AR vault facility. The AB Control Decision process could be used to determine the applicability and adequacy of existing AB controls as well as any new controls that may be needed for the identified hazardous conditions associated with 244-AR vault flooding. This hazard evaluation does not constitute an accident analysis.

  16. DNA damage induction of ribonucleotide reductase.

    PubMed

    Elledge, S J; Davis, R W

    1989-11-01

    RNR2 encodes the small subunit of ribonucleotide reductase, the enzyme that catalyzes the first step in the pathway for the production of deoxyribonucleotides needed for DNA synthesis. RNR2 is a member of a group of genes whose activities are cell cycle regulated and that are transcriptionally induced in response to the stress of DNA damage. An RNR2-lacZ fusion was used to further characterize the regulation of RNR2 and the pathway responsible for its response to DNA damage. beta-Galactosidase activity in yeast strains containing the RNR2-lacZ fusion was inducible in response to DNA-damaging agents (UV light, 4-nitroquinoline-1-oxide [4-NQO], and methyl methanesulfonate [MMS]) and agents that block DNA replication (hydroxyurea [HU] and methotrexate) but not heat shock. When MATa cells were arrested in G1 by alpha-factor, RNR2 mRNA was still inducible by DNA damage, indicating that the observed induction can occur outside of S phase. In addition, RNR2 induction was not blocked by the presence of cycloheximide and is therefore likely to be independent of protein synthesis. A mutation, rnr2-314, was found to confer hypersensitivity to HU and increased sensitivity to MMS. In rnr2-314 mutant strains, the DNA damage stress response was found to be partially constitutive as well as hypersensitive to induction by HU but not MMS. The induction properties of RNR2 were examined in a rad4-2 mutant background; in this genetic background, RNR2 was hypersensitive to induction by 4-NQO but not MMS. Induction of the RNR2-lacZ fusion in a RAD(+) strain in response to 4-NQO was not enhanced by the presence of an equal number of rad4-2 cells that lacked the fusion, implying that the DNA damage stress response in cell autonomous. PMID:2513480

  17. SUMO-specific protease 1 modulates cadmium-augmented transcriptional activity of androgen receptor (AR) by reversing AR SUMOylation.

    PubMed

    Wu, Ruiqin; Cui, Yaxiong; Yuan, Xiaoyan; Yuan, Haitao; Wang, Yimei; He, Jun; Zhao, Jun; Peng, Shuangqing

    2014-09-01

    Cadmium is a potential prostate carcinogen and can mimic the effects of androgen by a mechanism that involves the hormone-binding domain of the androgen receptor (AR), which is a key transcriptional factor in prostate carcinogenesis. We focused on transcriptional activity of AR to investigate the toxicity of cadmium exposure on human prostate cell lines. Cadmium increased the proliferative index of LNCaP and the proliferative effect was obstructed significantly by AR blocking agent. In luciferase assay, cadmium activated the transcriptional activity of AR in 293T cells co-transfected with wild-type AR and an ARE (AR response elements)-luciferase reporter gene. Cadmium also increased expression of PSA, a downstream gene of AR, whereas the metal had no significant effect on AR amount. AR is regulated by multiple posttranslational modifications including SUMOylation. SUMOylated AR shows a lower transcriptional activity. SUMO-specific protease 1 (SENP1) decreases AR SUMOylation by deconjugating AR-SUMO covalent bond. We detected that cadmium increased the amount of SENP1 in a dose and time dependent manner. Knocking down of SENP1 by RNAi led to decrease of PSA expression and transcriptional activity of AR in luciferase assay. Furthermore, co-immunoprecipitation (Co-IP) results showed that SUMOylation level of AR was decreased after cadmium treatment. In conclusion, our results indicated that cadmium-induced SENP1 enhanced AR transcriptional activity by decreasing AR SUMOylation.

  18. 40Ar/39Ar Age Dating of Two Chondrules from the Bjurböle (L/LL4) Chondrite

    NASA Astrophysics Data System (ADS)

    Park, J.; Lindsay, F. N.; Turrin, B. D.; Herzog, G. F.; Delaney, J. S.; Swisher, C. C.

    2016-08-01

    We present detailed 40Ar/39Ar results for two Bjurböle chondrules. Younger Ar/Ar step ages for one Bjurböle chondrule suggest that the analysis of single chondrules can yield more accurate meteorite ages than are obtainable from bulk samples.

  19. Mobility of Ar+ in CF4

    NASA Astrophysics Data System (ADS)

    Nikitovic, Zeljka; Stojanovic, Vladimir; Raspopovic, Zoran; Jovanovic, Jasmina; Petrovic, Zoran Lj.

    2015-09-01

    In this work we present a complete cross section set for Ar+ in CF4 where existing experimentally obtained data are selected and extrapolated. Monte Carlo simulation method is applied to accurately calculate transport parameters in hydrodynamic regime. We discuss new data for Ar+ ions in CF4 where flux and bulk values of reduced mobility are given as a function of E/N (E-electric field, N-gas density). We find that internally resonant exothermic dissociative charge transfer cross section for CF3+production significantly increases zero field ion mobility with respect to the polarization limit.

  20. Ar-Ar and I-Xe Ages and the Thermal History of IAB Meteorites

    NASA Technical Reports Server (NTRS)

    Bogard, Donald D.; Garrison, Daniel H.; Takeda, Hiroshi

    2005-01-01

    Studies of several samples of the large Caddo County IAB iron meteorite reveal andesitic material, enriched in Si, Na, Al and Ca, which is essentially unique among meteorites. This material is believed to have formed from a chondritic source by partial melting and to have further segregated by grain coarsening. Such an origin implies extended metamorphism of the IAB parent body. New Ar-39- Ar-40 ages for silicate from three different Caddo samples are consistent with a common age of 4.50-4.51 Gyr ago. Less well defined Ar-Ar degassing ages for inclusions from two other IABs, EET8333 and Udei Station, are approx.4.32 Gyr, whereas the age for Campo del Cielo varies considerably over approx.3.23-4.56 Gyr. New I-129-Xe-129 ages for Caddo County and EET8333 are 4557.9+/-0.1 Myr and 4557-4560 Myr, respectively, relative to an age of 4562.3 Myr for Shallowater. Considering all reported Ar-Ar degassing ages for IABs and related winonaites, the range is approx.4.32-4.53 Gyr, but several IABs give similar Ar ages of 4.50-4.52 Gyr. We interpret these older Ar ages to represent cooling after the time of last significant metamorphism on the parent body, and the younger ages to represent later 40Ar diffusion loss. The older Ar-Ar ages for IABs are similar to Sm-Nd and Rb-Sr isochron ages reported in the literature for Caddo County. Considering the possibility that IAB parent body formation was followed by impact disruption, reassembly, and metamorphism (e.g., Benedix et al. 2000), the Ar-Ar ages and IAB cooling rates deduced from Ni concentration profiles in IAB metal (Herpfer et al., 1994) are consistent if the time of the post-assembly metamorphism was as late as approx.4.53 Gyr ago. However, I-Xe ages reported for some IABs define much older ages of approx.4558-4566 Myr, which cannot easily be reconciled with the much younger Ar-Ar and Sm-Nd ages. An explanation for the difference in radiometric ages of IABs may reside in combinations of the following: a) I-Xe ages have very

  1. Partial vinylphenol reductase purification and characterization from Brettanomyces bruxellensis.

    PubMed

    Tchobanov, Iavor; Gal, Laurent; Guilloux-Benatier, Michèle; Remize, Fabienne; Nardi, Tiziana; Guzzo, Jean; Serpaggi, Virginie; Alexandre, Hervé

    2008-07-01

    Brettanomyces is the major microbial cause for wine spoilage worldwide and causes significant economic losses. The reasons are the production of ethylphenols that lead to an unpleasant taint described as 'phenolic odour'. Despite its economic importance, Brettanomyces has remained poorly studied at the metabolic level. The origin of the ethylphenol results from the conversion of vinylphenols in ethylphenol by Brettanomyces hydroxycinnamate decarboxylase. However, no information is available on the vinylphenol reductase responsible for the conversion of vinylphenols in ethylphenols. In this study, a vinylphenol reductase was partially purified from Brettanomyces bruxellensis that was active towards 4-vinylguaiacol and 4-vinylphenol only among the substrates tested. First, a vinylphenol reductase activity assay was designed that allowed us to show that the enzyme was NADH dependent. The vinylphenol reductase was purified 152-fold with a recovery yield of 1.77%. The apparent K(m) and V(max) values for the hydrolysis of 4-vinylguaiacol were, respectively, 0.14 mM and 1900 U mg(-1). The optimal pH and temperature for vinylphenol reductase were pH 5-6 and 30 degrees C, respectively. The molecular weight of the enzyme was 26 kDa. Trypsic digest of the protein was performed and the peptides were sequenced, which allowed us to identify in Brettanomyces genome an ORF coding for a 210 amino acid protein.

  2. Study of Ar and Ar-CO2 microwave surfaguide discharges by optical spectroscopy

    NASA Astrophysics Data System (ADS)

    Silva, Tiago; Britun, Nikolay; Godfroid, Thomas; van der Mullen, Joost; Snyders, Rony

    2016-05-01

    A surfaguide microwave discharge operating at 2.45 GHz in Ar and Ar-CO2 mixtures is studied using diagnostics methods based on optical emission spectroscopy. The population densities of Ar metastable and resonant states of the lowest group of excited levels ( 1 s x ) are investigated for several experimental conditions using the self-absorption technique. It is found that the densities of these levels, ranging from 1017 to 1016 m-3 for the pure Ar case, are dependent on the discharge pressure and applied power. The electron temperature and electron density are calculated via the balances of creation/loss mechanisms of radiative and metastable levels. In the range of the studied experimental conditions (50-300 W of applied power and 0.5-6 Torr of gas pressure), the results have shown that lower values of electron temperature correspond to higher values of power and pressure in the discharge. Adding CO2 to the argon plasma results in a considerable decrease (about 3 orders of magnitude) of the Ar metastable atom density. The feasibility of using the ratio of two Ar emission line intensities to measure the electron temperature in CO2 discharges with small Ar admixtures is studied.

  3. Ar-40/Ar-39 Ages of Maskelynite Grains from ALHA 77005

    NASA Technical Reports Server (NTRS)

    Turrin, B.; Park, J.; Herzog, G. F.; Lindsay, F. N.; Delaney, J. S.; Nyquist, L. E.; Swisher, C., III

    2013-01-01

    We present Ar-40/Ar-39 measurements for twelve small (20-60 micro-g) maskelynite samples from the heavily shocked martian meteorite ALHA 77005. The reported modal composition for ALHA 77005 is 50-60% olivine (Fa28), 30-40% pyroxene (Wo5Fs23En72), approx.8% maskelynite (An53), and approx.2% opaques by volume [1]). The meteorite is usually classified as a lherzolite. Previous Studies - Ar-40/Ar-39 results from previous work display disturbed release spectra [2,3]. In study [2], Ar-40/Ar-39 measurements on a 52-mg whole-rock sample produced an extremely disturbed release spec-trum, with all calculated apparent ages > 1 Ga, (Fig. 1). In a subsequent study [3], a light and a dark phase were analyzed. A 2.3-mg sample of the light, relatively low-K phase produced a disturbed release spectrum. For the first 20% of the Ar-39(sub K), most of the apparent ages exceeded >1 Ga; the remaining 80% yielded ages between 0.3-0.5 Ga. The integrated age for this phase is 0.9 Ga.

  4. Ar-Ar Impact Heating Ages of Eucrites and Timing of the LHB

    NASA Technical Reports Server (NTRS)

    Bogard, Donald; Garrison, Daniel

    2009-01-01

    Eucrites and howardites, more than most meteorite types, show extensive impact resetting of their Ar-39-Ar-40 (K-Ar) ages approximately equal to 3.4-4.1 Ga ago, and many specimens show some disturbance of other radiometry chronometers as well. Bogard (1995) argued that this age resetting occurred on Vesta and was produced by the same general population of objects that produced many of the lunar impact basins. The exact nature of the lunar late heavy bombardment (LHB or 'cataclysm') remains controversial, but the timing is similar to the reset ages of eucrites. Neither the beginning nor ending time of the lunar LHB is well constrained. Comparison of Ar-Ar ages of brecciated eucrites with data for the lunar LHB can resolve both the origin of these impactors and the time period over which they were delivered to the inner solar system. This abstract reports some new Ar-Ar age data for eucrites, obtained since the authors' 1995 and 2003 papers.

  5. Moving Beyond the Androgen Receptor (AR): Targeting AR-Interacting Proteins to Treat Prostate Cancer.

    PubMed

    Foley, Christopher; Mitsiades, Nicholas

    2016-04-01

    Medical or surgical castration serves as the backbone of systemic therapy for advanced and metastatic prostate cancer, taking advantage of the importance of androgen signaling in this disease. Unfortunately, resistance to castration emerges almost universally. Despite the development and approval of new and more potent androgen synthesis inhibitors and androgen receptor (AR) antagonists, prostate cancers continue to develop resistance to these therapeutics, while often maintaining their dependence on the AR signaling axis. This highlights the need for innovative therapeutic approaches that aim to continue disrupting AR downstream signaling but are orthogonal to directly targeting the AR itself. In this review, we discuss the preclinical research that has been done, as well as clinical trials for prostate cancer, on inhibiting several important families of AR-interacting proteins, including chaperones (such as heat shock protein 90 (HSP90) and FKBP52), pioneer factors (including forkhead box protein A1 (FOXA1) and GATA-2), and AR transcriptional coregulators such as the p160 steroid receptor coactivators (SRCs) SRC-1, SRC-2, SRC-3, as well as lysine deacetylases (KDACs) and lysine acetyltransferases (KATs). Researching the effect of-and developing new therapeutic agents that target-the AR signaling axis is critical to advancing our understanding of prostate cancer biology, to continue to improve treatments for prostate cancer and for overcoming castration resistance.

  6. Enhanced 2,3-Butanediol Production by Optimizing Fermentation Conditions and Engineering Klebsiella oxytoca M1 through Overexpression of Acetoin Reductase

    PubMed Central

    Cho, Sukhyeong; Kim, Taeyeon; Woo, Han Min; Lee, Jinwon; Kim, Yunje; Um, Youngsoon

    2015-01-01

    Microbial production of 2,3-butanediol (2,3-BDO) has been attracting increasing interest because of its high value and various industrial applications. In this study, high production of 2,3-BDO using a previously isolated bacterium Klebsiella oxytoca M1 was carried out by optimizing fermentation conditions and overexpressing acetoin reductase (AR). Supplying complex nitrogen sources and using NaOH as a neutralizing agent were found to enhance specific production and yield of 2,3-BDO. In fed-batch fermentations, 2,3-BDO production increased with the agitation speed (109.6 g/L at 300 rpm vs. 118.5 g/L at 400 rpm) along with significantly reduced formation of by-product, but the yield at 400 rpm was lower than that at 300 rpm (0.40 g/g vs. 0.34 g/g) due to acetoin accumulation at 400 rpm. Because AR catalyzing both acetoin reduction and 2,3-BDO oxidation in K. oxytoca M1 revealed more than 8-fold higher reduction activity than oxidation activity, the engineered K. oxytoca M1 overexpressing the budC encoding AR was used in fed-batch fermentation. Finally, acetoin accumulation was significantly reduced by 43% and enhancement of 2,3-BDO concentration (142.5 g/L), yield (0.42 g/g) and productivity (1.47 g/L/h) was achieved compared to performance with the parent strain. This is by far the highest titer of 2,3-BDO achieved by K. oxytoca strains. This notable result could be obtained by finding favorable fermentation conditions for 2,3-BDO production as well as by utilizing the distinct characteristic of AR in K. oxytoca M1 revealing the nature of reductase. PMID:26368397

  7. Combined effect of polymorphisms in type III 5-α reductase and androgen receptor gene with the risk of benign prostatic hyperplasia in Korea

    PubMed Central

    Lee, Chung Lyul; Lee, Jaegeun; Na, Yong Gil; Song, Ki Hak

    2016-01-01

    We evaluated whether type III 5-alpha reductase (SRD5A3; steroid reductase 5-alpha 3) polymorphism was associated with susceptibility of benign prostate hyperplasia (BPH) and the combined effects in BPH risk between the type of short tandem repeat (STR) in SRD5A3 and the length of trinucleotide (CAG) repeats in androgen receptor (AR) gene. We compared the length of AC repeats in STR region of SRD5A3 gene and a CAG repeat in AR in 188 BPH patients who underwent transurethral resection of prostate (TURP) and 98 controls by polymerase chain reaction-based methods. We defined short type was less than 21 copies of AC repeats. The odds ratio for BPH between the men with at least one of short type and with both large types of STR in SRD5A3 gene was 3.10 (95% confidence interval [CI], 1.87–5.16; P=0.000). And BPH was 2.35 times more likely to occur in with less than 23 copies of CAG repeats than men equal or greater than 23 copies in AR gene (95% CI, 1.18–2.36; P=0.016). The men with the large type of STR and ≥23 copies of CAG repeats have 5.3 times BPH risk compared to the reference group with the at least one of the short type of STR and <23 copies (P<0.000). In conclusion, these results suggest that shorter AC repeats of SRD5A3 gene and shorter CAG repeats of AR gene were associated with an increased risk for BPH. However, the interaction between above two factors was not affected in risk of BPH. PMID:27807533

  8. Potassium Isotopic Compositions of NIST Potassium Standards and 40Ar/39Ar Mineral Standards

    NASA Technical Reports Server (NTRS)

    Morgan, Leah; Tappa, Mike; Ellam, Rob; Mark, Darren; Higgins, John; Simon, Justin I.

    2013-01-01

    Knowledge of the isotopic ratios of standards, spikes, and reference materials is fundamental to the accuracy of many geochronological methods. For example, the 238U/235U ratio relevant to U-Pb geochronology was recently re-determined [1] and shown to differ significantly from the previously accepted value employed during age determinations. These underlying values are fundamental to accurate age calculations in many isotopic systems, and uncertainty in these values can represent a significant (and often unrecognized) portion of the uncertainty budget for determined ages. The potassium isotopic composition of mineral standards, or neutron flux monitors, is a critical, but often overlooked component in the calculation of K-Ar and 40Ar/39Ar ages. It is currently assumed that all terrestrial materials have abundances indistinguishable from that of NIST SRM 985 [2]; this is apparently a reasonable assumption at the 0.25per mille level (1s) [3]. The 40Ar/39Ar method further relies on the assumption that standards and samples (including primary and secondary standards) have indistinguishable 40K/39K values. We will present data establishing the potassium isotopic compositions of NIST isotopic K SRM 985, elemental K SRM 999b, and 40Ar/39Ar biotite mineral standard GA1550 (sample MD-2). Stable isotopic compositions (41K/39K) were measured by the peak shoulder method with high resolution MC-ICP-MS (Thermo Scientific NEPTUNE Plus), using the accepted value of NIST isotopic SRM 985 [2] for fractionation [4] corrections [5]. 40K abundances were measured by TIMS (Thermo Scientific TRITON), using 41K/39K values from ICP-MS measurements (or, for SRM 985, values from [2]) for internal fractionation corrections. Collectively these data represent an important step towards a metrologically traceable calibration of 40K concentrations in primary 40Ar/39Ar mineral standards and improve uncertainties by ca. an order of magnitude in the potassium isotopic compositions of standards.

  9. Whole-Rock 40Ar/39Ar Step-heating Analyses, Problems and Potential

    NASA Astrophysics Data System (ADS)

    Boehnke, P.; Harrison, M.; Heizler, M. T.; Lovera, O. M.; Warren, P. H.

    2015-12-01

    Whole-rock 40Ar/39Ar step-heating analyses of extra-terrestrial materials are used to constrain the impact history of the inner solar system, the formation age of the Moon, and timing of paleomagnetic fields. Despite the importance of knowing the timing of these important events, the samples we have in hand are usually disturbed through mixing, (multiple?) impact events, and perhaps recoil loss. Extra-terrestrial 40Ar/39Ar data are typically interpreted through the assignment of essentially arbitrary plateau ages rather than through a robust physical model. Although the use of models capable of quantitatively assessing diffusive 40Ar* loss in extra-terrestrial samples has been around for nearly 50 years, this early advance has been widely ignored. Here we present implications of applying a robust, multi-activation energy, multi-diffusion domain model to step-heated 40Ar/39Ar data, with temperature cycling. Our findings show that for even a single heating event, "plateau" ages are unlikely to record meaningful ages. Further, if the sample has experienced multiple heating events or contains inherited clasts, recovering a unique solution may be impossible. Indeed the most readily interpretable portion of the age spectrum is the early heating steps which represents a maximum age estimate of the last re-heating event. Our results challenge the chronologic validity of 40Ar/39Ar "plateau" ages and by extension the hypotheses that are based on this data (e.g., the Late Heavy Bombardment). Future work will require new analytical procedures, interpretative frameworks, and (potentially) the combination of multiple chronometers to derive a robust impact history for the early solar system.

  10. Experiences from the ARS croplands CEAP program

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The multi-agency Conservation Effects Assessment Project (CEAP) within USDA produced a number of lessons that should be applicable to the use of landscape approaches to place bioenergy crops. Results from the ARS Croplands Watersheds CEAP, the NRCS CEAP, and the NIFA CEAP Watershed Assessment Studie...

  11. "Ars Dictaminis" and Modern Rhetorical Practice.

    ERIC Educational Resources Information Center

    Sewell, Ernestine P.

    Although not all the problems of writing are solved by practice in letter writing, a review of the classical texts on "ars dictaminis"--letter writing--creates a strong argument for its increased use as an exercise in college composition classrooms. By incorporating the lessons of dictamen as a device for achieving rhetorical modes--narration,…

  12. 75 FR 7637 - Arkansas Disaster #AR-00040

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-22

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00040 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a notice of an Administrative declaration of a disaster for the State of Arkansas dated...

  13. 76 FR 27140 - Arkansas Disaster # AR-00049

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-10

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00049 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a Notice of the Presidential declaration of a major disaster for Public Assistance Only for...

  14. 75 FR 7636 - Arkansas Disaster #AR-00042

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-22

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00042 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a Notice of the Presidential declaration of a major disaster for Public Assistance Only for...

  15. 76 FR 42154 - Arkansas Disaster #AR-00050

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-18

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00050 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a Notice of the Presidential declaration of a major disaster for the State of Arkansas...

  16. 78 FR 39821 - Arkansas Disaster #AR-00064

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-02

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00064 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a Notice of the Presidential declaration of a major disaster for Public Assistance Only for...

  17. 76 FR 42155 - Arkansas Disaster #AR-00051

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-18

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00051 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a Notice of the Presidential declaration of a major disaster for Public Assistance Only for...

  18. 75 FR 30872 - Arkansas Disaster # AR-00043

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-02

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00043 AGENCY: U.S. Small Business Administration. ACTION: Notice SUMMARY: This is a notice of an Administrative declaration of a disaster for the State of Arkansas dated...

  19. 78 FR 56979 - Arkansas Disaster # AR-00065

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-09-16

    ... From the Federal Register Online via the Government Publishing Office ] SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00065 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a Notice of the Presidential declaration of a major disaster for Public Assistance Only for...

  20. 76 FR 27139 - Arkansas Disaster #AR-00048

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-10

    ... From the Federal Register Online via the Government Publishing Office ] SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00048 AGENCY: U.S. Small Business Administration. ACTION: Notice. SUMMARY: This is a Notice of the Presidential declaration of a major disaster for the State of Arkansas...

  1. 78 FR 9448 - Arkansas Disaster #AR-00061

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-08

    ... From the Federal Register Online via the Government Publishing Office SMALL BUSINESS ADMINISTRATION Arkansas Disaster AR-00061 AGENCY: U.S. Small Business Administration. ACTION: Notice SUMMARY: This is a Notice of the Presidential declaration of a major disaster for Public Assistance Only for...

  2. The ChArMEx database

    NASA Astrophysics Data System (ADS)

    Ferré, Hélène; Belmahfoud, Nizar; Boichard, Jean-Luc; Brissebrat, Guillaume; Cloché, Sophie; Descloitres, Jacques; Fleury, Laurence; Focsa, Loredana; Henriot, Nicolas; Mière, Arnaud; Ramage, Karim; Vermeulen, Anne; Boulanger, Damien

    2015-04-01

    The Chemistry-Aerosol Mediterranean Experiment (ChArMEx, http://charmex.lsce.ipsl.fr/) aims at a scientific assessment of the present and future state of the atmospheric environment in the Mediterranean Basin, and of its impacts on the regional climate, air quality, and marine biogeochemistry. The project includes long term monitoring of environmental parameters , intensive field campaigns, use of satellite data and modelling studies. Therefore ChARMEx scientists produce and need to access a wide diversity of data. In this context, the objective of the database task is to organize data management, distribution system and services, such as facilitating the exchange of information and stimulating the collaboration between researchers within the ChArMEx community, and beyond. The database relies on a strong collaboration between ICARE, IPSL and OMP data centers and has been set up in the framework of the Mediterranean Integrated Studies at Regional And Locals Scales (MISTRALS) program data portal. ChArMEx data, either produced or used by the project, are documented and accessible through the database website: http://mistrals.sedoo.fr/ChArMEx. The website offers the usual but user-friendly functionalities: data catalog, user registration procedure, search tool to select and access data... The metadata (data description) are standardized, and comply with international standards (ISO 19115-19139; INSPIRE European Directive; Global Change Master Directory Thesaurus). A Digital Object Identifier (DOI) assignement procedure allows to automatically register the datasets, in order to make them easier to access, cite, reuse and verify. At present, the ChArMEx database contains about 120 datasets, including more than 80 in situ datasets (2012, 2013 and 2014 summer campaigns, background monitoring station of Ersa...), 25 model output sets (dust model intercomparison, MEDCORDEX scenarios...), a high resolution emission inventory over the Mediterranean... Many in situ datasets

  3. Crystal structure of red chlorophyll catabolite reductase: enlargement of the ferredoxin-dependent bilin reductase family.

    PubMed

    Sugishima, Masakazu; Kitamori, Yuka; Noguchi, Masato; Kohchi, Takayuki; Fukuyama, Keiichi

    2009-06-01

    The key steps in the degradation pathway of chlorophylls are the ring-opening reaction catalyzed by pheophorbide a oxygenase and sequential reduction by red chlorophyll catabolite reductase (RCCR). During these steps, chlorophyll catabolites lose their color and phototoxicity. RCCR catalyzes the ferredoxin-dependent reduction of the C20/C1 double bond of red chlorophyll catabolite. RCCR appears to be evolutionarily related to the ferredoxin-dependent bilin reductase (FDBR) family, which synthesizes a variety of phytobilin pigments, on the basis of sequence similarity, ferredoxin dependency, and the common tetrapyrrole skeleton of their substrates. The evidence, however, is not robust; the identity between RCCR and FDBR HY2 from Arabidopsis thaliana is only 15%, and the oligomeric states of these enzymes are different. Here, we report the crystal structure of A. thaliana RCCR at 2.4 A resolution. RCCR forms a homodimer, in which each subunit folds in an alpha/beta/alpha sandwich. The tertiary structure of RCCR is similar to those of FDBRs, strongly supporting that these enzymes evolved from a common ancestor. The two subunits are related by noncrystallographic 2-fold symmetry in which the alpha-helices near the edge of the beta-sheet unique in RCCR participate in intersubunit interaction. The putative RCC-binding site, which was derived by superimposing RCCR onto biliverdin-bound forms of FDBRs, forms an open pocket surrounded by conserved residues among RCCRs. Glu154 and Asp291 of A. thaliana RCCR, which stand opposite each other in the pocket, likely are involved in substrate binding and/or catalysis.

  4. 4-Dimethylaminoazobenzenes: carcinogenicities and reductive cleavage by microsomal azo reductase.

    PubMed

    Lambooy, J P; Koffman, B M

    1985-01-01

    Twenty-four 4-dimethylaminoazobenzenes (DABs) in which systematic structural modifications have been made in the prime ring have been studied for substrate specificity for microsomal azo reductase. The DABs were also evaluated for carcinogenicity and it was found that there was no correlation between carcinogenicity and extent of azo bond cleavage by azo reductase. While any substituent in the prime ring reduces the rate of cleavage of the azo bond relative to the unsubstituted dye, there is a correlation between substituent size and susceptibility to the enzyme. Substituent size was also found to be a significant factor in the induction of hepatomas by the dyes. Preliminary studies have shown that there appears to be a positive correlation between microsomal riboflavin content and the activity of the azo reductase.

  5. Sulfur Isotope Effects of Dissimilatory Sulfite Reductase.

    PubMed

    Leavitt, William D; Bradley, Alexander S; Santos, André A; Pereira, Inês A C; Johnston, David T

    2015-01-01

    The precise interpretation of environmental sulfur isotope records requires a quantitative understanding of the biochemical controls on sulfur isotope fractionation by the principle isotope-fractionating process within the S cycle, microbial sulfate reduction (MSR). Here we provide the only direct observation of the major ((34)S/(32)S) and minor ((33)S/(32)S, (36)S/(32)S) sulfur isotope fractionations imparted by a central enzyme in the energy metabolism of sulfate reducers, dissimilatory sulfite reductase (DsrAB). Results from in vitro sulfite reduction experiments allow us to calculate the in vitro DsrAB isotope effect in (34)S/(32)S (hereafter, [Formula: see text]) to be 15.3 ± 2‰, 2σ. The accompanying minor isotope effect in (33)S, described as [Formula: see text], is calculated to be 0.5150 ± 0.0012, 2σ. These observations facilitate a rigorous evaluation of the isotopic fractionation associated with the dissimilatory MSR pathway, as well as of the environmental variables that govern the overall magnitude of fractionation by natural communities of sulfate reducers. The isotope effect induced by DsrAB upon sulfite reduction is a factor of 0.3-0.6 times prior indirect estimates, which have ranged from 25 to 53‰ in (34)εDsrAB. The minor isotope fractionation observed from DsrAB is consistent with a kinetic or equilibrium effect. Our in vitro constraints on the magnitude of [Formula: see text] is similar to the median value of experimental observations compiled from all known published work, where (34)ε r-p = 16.1‰ (r-p indicates reactant vs. product, n = 648). This value closely matches those of MSR operating at high sulfate reduction rates in both laboratory chemostat experiments ([Formula: see text] 17.3 ± 1.5‰, 2σ) and in modern marine sediments ([Formula: see text] 17.3 ± 3.8‰). Targeting the direct isotopic consequences of a specific enzymatic processes is a fundamental step toward a biochemical foundation for reinterpreting the

  6. Sulfur Isotope Effects of Dissimilatory Sulfite Reductase

    PubMed Central

    Leavitt, William D.; Bradley, Alexander S.; Santos, André A.; Pereira, Inês A. C.; Johnston, David T.

    2015-01-01

    The precise interpretation of environmental sulfur isotope records requires a quantitative understanding of the biochemical controls on sulfur isotope fractionation by the principle isotope-fractionating process within the S cycle, microbial sulfate reduction (MSR). Here we provide the only direct observation of the major (34S/32S) and minor (33S/32S, 36S/32S) sulfur isotope fractionations imparted by a central enzyme in the energy metabolism of sulfate reducers, dissimilatory sulfite reductase (DsrAB). Results from in vitro sulfite reduction experiments allow us to calculate the in vitro DsrAB isotope effect in 34S/32S (hereafter, 34εDsrAB) to be 15.3 ± 2‰, 2σ. The accompanying minor isotope effect in 33S, described as 33λDsrAB, is calculated to be 0.5150 ± 0.0012, 2σ. These observations facilitate a rigorous evaluation of the isotopic fractionation associated with the dissimilatory MSR pathway, as well as of the environmental variables that govern the overall magnitude of fractionation by natural communities of sulfate reducers. The isotope effect induced by DsrAB upon sulfite reduction is a factor of 0.3–0.6 times prior indirect estimates, which have ranged from 25 to 53‰ in 34εDsrAB. The minor isotope fractionation observed from DsrAB is consistent with a kinetic or equilibrium effect. Our in vitro constraints on the magnitude of 34εDsrAB is similar to the median value of experimental observations compiled from all known published work, where 34εr−p = 16.1‰ (r–p indicates reactant vs. product, n = 648). This value closely matches those of MSR operating at high sulfate reduction rates in both laboratory chemostat experiments (34εSO4−H2S =  17.3 ± 1.5‰, 2σ) and in modern marine sediments (34εSO4−H2S =  17.3 ± 3.8‰). Targeting the direct isotopic consequences of a specific enzymatic processes is a fundamental step toward a biochemical foundation for reinterpreting the biogeochemical and geobiological sulfur isotope records in

  7. Mechanistic imperatives for aldose-ketose isomerization in water: specific, general base- and metal ion-catalyzed isomerization of glyceraldehyde with proton and hydride transfer.

    PubMed

    Nagorski, R W; Richard, J P

    2001-02-01

    The deuterium enrichment of dihydroxyacetone obtained from the aldose-ketose isomerization of D,L-glyceraldehyde in D(2)O at 25 degrees C was determined by (1)H NMR spectroscopy from the integrated areas of the signals for the alpha-CH(2) and alpha-CHD groups of the product. One mole equivalent of deuterium is incorporated into the product when the isomerization is carried out in 150 mM pyrophosphate buffer at pD 8.4, but only 0.6 mol equiv of deuterium is incorporated into the product of isomerization in the presence of 0.01 M deuterioxide ion, so that 40% of the latter isomerization reaction proceeds by the intramolecular transfer of hydride ion. Several pathways were identified for catalysis of the isomerization of glyceraldehyde to give dihydroxyacetone. The isomerization with hydride transfer is strongly catalyzed by added Zn(2+). Deprotonation of glyceraldehyde is rate-determining for isomerization with proton transfer, and this proton-transfer reaction is catalyzed by Brønsted bases. Proton transfer also occurs by a termolecular pathway with catalysis by the combined action of Brønsted bases and Zn(2+). These results show that there is no large advantage to the spontaneous isomerization of glyceraldehyde in alkaline solution with either proton or hydride transfer, and that effective catalytic pathways exist to stabilize the transition states for both of these reactions in water. The existence of separate enzymes that catalyze the isomerization of sugars with hydride transfer and the isomerization of sugar phosphates with proton transfer is proposed to be a consequence of the lack of any large advantage to reaction by either of these pathways for the corresponding nonenzymatic isomerization in water.

  8. Age and origin of carlsbad cavern and related caves from 40Ar/39Ar of alunite

    PubMed

    Polyak; McIntosh; Guven; Provencio

    1998-03-20

    40Ar/39Ar dating of fine-grained alunite that formed during cave genesis provides ages of formation for the Big Room level of Carlsbad Cavern [4.0 to 3.9 million years ago (Ma)], the upper level of Lechuguilla Cave (6.0 to 5.7 Ma), and three other hypogene caves (11.3 to 6.0 Ma) in the Guadalupe Mountains of New Mexico. Alunite ages increase and are strongly correlative with cave elevations, which indicates an 1100-meter decline in the water table, apparently related to tectonic uplift and tilting, from 11.3 Ma to the present. 40Ar/39Ar dating studies of the hypogene caves have the potential to help resolve late Cenozoic climatic, speleologic, and tectonic questions.

  9. Planar defects as Ar traps in trioctahedral micas: A mechanism for increased Ar retentivity in phlogopite

    NASA Astrophysics Data System (ADS)

    Camacho, A.; Lee, J. K. W.; Fitz Gerald, J. D.; Zhao, J.; Abdu, Y. A.; Jenkins, D. M.; Hawthorne, F. C.; Kyser, T. K.; Creaser, R. A.; Armstrong, R.; Heaman, L. W.

    2012-08-01

    The effects of planar defects and composition on Ar mobility in trioctahedral micas have been investigated in samples from a small marble outcrop (∼500 m2) in the Frontenac Terrane, Grenville Province, Ontario. These micas crystallized during amphibolite-facies metamorphism at ∼1170 Ma and experienced a thermal pulse ∼100 Ma later at shallow crustal levels associated with the emplacement of plutons. 87Rb/86Sr ages of the phlogopites range from ∼950 to ∼1050 Ma, consistent with resetting during the later thermal event. The same phlogopites however, give 40Ar/39Ar ages between ∼950 and 1160 Ma, spanning the age range of the two thermal events. This result is intriguing because these micas have undergone the same thermal history and were not deformed after peak metamorphic conditions. In order to understand this phenomenon, the chemical, crystallographical, and microstructural nature of four mica samples has been characterized in detail using a wide range of analytical techniques. The scanning electron microscope (SEM), electron microprobe (EMP), and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) data show that the micas are chemically homogeneous (with the exception of Ba) and similar in composition. The Fourier transform infrared spectroscopy and Mossbauer results show that the M sites for three of the micas are dominated by divalent cations and the Fe3+/(Fe2++Fe3+) ratio for all four phlogopites ranges from 0.10 to 0.25. The stable-isotopic data for calcite indicate that this outcrop was not affected by hydrothermal fluids after peak metamorphism. No correlation between chemical composition and 87Rb/86Sr and 40Ar/39Ar age or between crystal size and 40Ar/39Ar age is observed. The only major difference among all of the micas was revealed through transmitted electron microscope (TEM), which shows that the older 1M micas contain significantly more layer stacking defects, associated with crystallization, than the younger micas. We

  10. A role for selective androgen response elements in the development of the epididymis and the androgen control of the 5α reductase II gene.

    PubMed

    Kerkhofs, Stefanie; Dubois, Vanessa; De Gendt, Karel; Helsen, Christine; Clinckemalie, Liesbeth; Spans, Lien; Schuit, Frans; Boonen, Steven; Vanderschueren, Dirk; Saunders, Philippa T K; Verhoeven, Guido; Claessens, Frank

    2012-10-01

    The androgen receptor (AR) recognizes two types of DNA elements that are dimers of 5'-AGAACA-3'-like hexamers, either organized as inverted or direct repeats. We developed a mouse model [(specificity affecting AR knock-in (SPARKI)] in which the AR DNA-binding domain was mutated such that it lost binding to direct repeats but not to inverted elements. The impaired fertility of the male SPARKI mice correlates with the reduced motility of the spermatozoa, a characteristic that is developed during transit through the epididymis. Comparative transcriptome analyses revealed that the expression of 39 genes is changed in SPARKI epididymis. Remarkably, the expression of the steroid 5α-reductase type II (Srd5α2) gene, which metabolizes testosterone into the more potent dihydrotestosterone, is reduced 4-fold in SPARKI vs. wild type. The comparison of the SPARKI phenotype with that of Srd5α2-knockout mice shows, however, that the reduced Srd5α2 expression cannot explain all defects of the SPARKI epididymis. Moreover, we describe three new selective androgen response elements (AREs), which control the androgen responsiveness of the Srd5α2 gene. We conclude that the SPARKI model can be considered a knockout model for AR functioning via selective AREs and that this has a dramatic effect on sperm maturation in the epididymis.

  11. Molecular effects of bioactive fraction of Curcuma mangga (DLBS4847) as a downregulator of 5α-reductase activity pathways in prostatic epithelial cells.

    PubMed

    Karsono, Agung Heru; Tandrasasmita, Olivia Mayasari; Tjandrawinata, Raymond R

    2014-01-01

    DLBS4847 is a standardized bioactive fraction of Curcuma mangga. In this study, we used prostate cancer (PC)-3 as the cell line to study the effects of DLBS4847 on prostatic cell viability, as well as related molecular changes associated with the decreased cell number. The observation revealed that DLBS4847 inhibited the growth of PC3 cells through downregulation of the 5α-reductase (5AR) pathway. At the transcription level, 5AR1 and androgen-receptor gene expressions were downregulated in a dose-dependent manner. Furthermore, 5AR-1 and dihydrotestosterone expression were also downregulated at the protein level. A microarray study was also performed to see the effects of DLBS4847 on differential gene expressions in prostate cancer 3 cells. Among others, DLBS4847 downregulated genes related to prostate growth and hypertrophy. Our results suggested that DLBS4847 could potentially become an alternative treatment for prostate disorders, such as benign prostatic hyperplasia. In this regard, DLBS4847 exerts its growth inhibition partially through downregulation of the 5AR pathway.

  12. Molecular effects of bioactive fraction of Curcuma mangga (DLBS4847) as a downregulator of 5α-reductase activity pathways in prostatic epithelial cells

    PubMed Central

    Karsono, Agung Heru; Tandrasasmita, Olivia Mayasari; Tjandrawinata, Raymond R

    2014-01-01

    DLBS4847 is a standardized bioactive fraction of Curcuma mangga. In this study, we used prostate cancer (PC)-3 as the cell line to study the effects of DLBS4847 on prostatic cell viability, as well as related molecular changes associated with the decreased cell number. The observation revealed that DLBS4847 inhibited the growth of PC3 cells through downregulation of the 5α-reductase (5AR) pathway. At the transcription level, 5AR1 and androgen-receptor gene expressions were downregulated in a dose-dependent manner. Furthermore, 5AR-1 and dihydrotestosterone expression were also downregulated at the protein level. A microarray study was also performed to see the effects of DLBS4847 on differential gene expressions in prostate cancer 3 cells. Among others, DLBS4847 downregulated genes related to prostate growth and hypertrophy. Our results suggested that DLBS4847 could potentially become an alternative treatment for prostate disorders, such as benign prostatic hyperplasia. In this regard, DLBS4847 exerts its growth inhibition partially through downregulation of the 5AR pathway. PMID:24944518

  13. Early Pleistocene 40Ar/39Ar ages for Bapang Formation hominins, Central Jawa, Indonesia

    PubMed Central

    Larick, Roy; Ciochon, Russell L.; Zaim, Yahdi; Sudijono; Suminto; Rizal, Yan; Aziz, Fachroel; Reagan, Mark; Heizler, Matthew

    2001-01-01

    The Sangiran dome is the primary stratigraphic window for the Plio-Pleistocene deposits of the Solo basin of Central Jawa. The dome has yielded nearly 80 Homo erectus fossils, around 50 of which have known findspots. With a hornblende 40Ar/39Ar plateau age of 1.66 ± 0.04 mega-annum (Ma) reportedly associated with two fossils [Swisher, C.C., III, Curtis, G. H., Jacob, T., Getty, A. G., Suprijo, A. & Widiasmoro (1994) Science 263, 1118–1121), the dome offers evidence that early Homo dispersed to East Asia during the earliest Pleistocene. Unfortunately, the hornblende pumice was sampled at Jokotingkir Hill, a central locality with complex lithostratigraphic deformation and dubious specimen provenance. To address the antiquity of Sangiran H. erectus more systematically, we investigate the sedimentary framework and hornblende 40Ar/39Ar age for volcanic deposits in the southeast quadrant of the dome. In this sector, Bapang (Kabuh) sediments have their largest exposure, least deformation, and most complete tephrostratigraphy. At five locations, we identify a sequence of sedimentary cycles in which H. erectus fossils are associated with epiclastic pumice. From sampled pumice, eight hornblende separates produced 40Ar/39Ar plateau ages ranging from 1.51 ± 0.08 Ma at the Bapang/Sangiran Formation contact, to 1.02 ± 0.06 Ma, at a point above the hominin-bearing sequence. The chronological sequence of 40Ar/39Ar ages follows stratigraphic order across the southeast quadrant. An intermediate level yielding four nearly complete crania has an age of about 1.25 Ma. PMID:11309488

  14. Ar-Ar Dating of Martian Meteorite, Dhofar 378: An Early Shock Event?

    NASA Technical Reports Server (NTRS)

    Park, J.; Bogard, D. D.

    2006-01-01

    Martian meteorite, Dhofar 378 (Dho378) is a basaltic shergottite from Oman, weighing 15 g, and possessing a black fusion crust. Chemical similarities between Dho378 and the Los Angeles 001 shergottite suggests that they might have derived from the same Mars locale. The plagioclase in other shergottites has been converted to maskelenite by shock, but Dho378 apparently experienced even more intense shock heating, estimated at 55-75 GPa. Dho378 feldspar (approximately 43 modal %) melted, partially flowed and vesiculated, and then partially recrystallized. Areas of feldspathic glass are appreciably enriched in K, whereas individual plagioclases show a range in the Or/An ratio of approximately 0.18-0.017. Radiometric dating of martian shergottites indicate variable formation times of 160-475 Myr, whereas cosmic ray exposure (CRE) ages of shergottites indicate most were ejected from Mars within the past few Myr. Most determined Ar-39-Ar-40 ages of shergottites appear older than other radiometric ages because of the presence of large amounts of martian atmosphere or interior Ar-40. Among all types of meteorites and returned lunar rocks, the impact event that initiated the CRE age very rarely reset the Ar-Ar age. This is because a minimum time and temperature is required to facilitate Ar diffusion loss. It is generally assumed that the shock-texture characteristics in martian meteorites were produced by the impact events that ejected the rocks from Mars, although the time of these shock events (as opposed to CRE ages) are not directly dated. Here we report Ar-39-Ar-40 dating of Dho378 plagioclase. We suggest that the determined age dates the intense shock heating event this meteorite experienced, but that it was not the impact that initiated the CRE age.

  15. Preliminary 40Ar-39Ar thermochronological study of Dien Bien Phu Fault, northern Vietnam

    NASA Astrophysics Data System (ADS)

    Lin, T.; Lo, C.; Wang, P.; Lee, T.; Chung, S.; Lan, C.

    2003-12-01

    The Song Ma belt, located along the suture zone between the South China and Indochina blocks, was offset by the Dien Bien Phu Fault (DBF) in northern Vietnam. Many previous studies suggested that the Dien Bien Phu Fault is one of the Tertiary shear zones, resulted from the Cenozoic extrusion tectonics due to the Eurasian-Indian collision. However, the timing of the movement of DBF is remained unclear due to lacking of age data. In the present study, schists and mylonitic granites from DBF, and undeformed granites from the granitic belt along the south China block, were analysed by 40Ar-39Ar method. K-feldspar, hornblende, and biotite samples form granite plutons along south edge of South China block displayed different age ranges between opposite sides of DBF. Samples from plutonic body which is located in west side exhibit plateau ages ranging form 207Ma to 229Ma. Whereas, hornblende separates from granites on the west side exhibit much older plateau dates at around 277Ma. More detail studies were required to get better understand about the discrepancy of dating results between granitic masses located in different sides of DBF. On the other hand, Laser 40Ar-39Ar single-grain analyses conducted on deformed biotites and muscovites separated from the schists in DBF, show 40Ar-39Ar ages ranging form 185Ma to 205Ma, which are much younger than the plateau dates of those undeformed granitic samples. In summary, K-feldspar, hornblende and biotite samples from the undeformed granites, exhibit plateau dates in the range of 207- 277Ma, reflecting the ages of plutonism during the Indosinian orogeny. Whereas, muscovites and biotites extracted from schists and deformed granites in DBF, display young 40Ar-39Ar ages in the range of 205-185 Ma. This may suggest that although the DBF might have been reactivated during the Cenozoic extrusion tectonics, but it was in fact active in the Mesozoic.

  16. Molecular characterization of Alr1105 a novel arsenate reductase of the diazotrophic cyanobacterium Anabaena sp. PCC7120 and decoding its role in abiotic stress management in Escherichia coli.

    PubMed

    Pandey, Sarita; Shrivastava, Alok K; Rai, Rashmi; Rai, Lal Chand

    2013-11-01

    This paper constitutes the first report on the Alr1105 of Anabaena sp. PCC7120 which functions as arsenate reductase and phosphatase and offers tolerance against oxidative and other abiotic stresses in the alr1105 transformed Escherichia coli. The bonafide of 40.8 kDa recombinant GST+Alr1105 fusion protein was confirmed by immunoblotting. The purified Alr1105 protein (mw 14.8 kDa) possessed strong arsenate reductase (Km 16.0 ± 1.2 mM and Vmax 5.6 ± 0.31 μmol min⁻¹ mg protein⁻¹) and phosphatase activity (Km 27.38 ± 3.1 mM and Vmax 0.077 ± 0.005 μmol min⁻¹ mg protein⁻¹) at an optimum temperature 37 °C and 6.5 pH. Native Alr1105 was found as a monomeric protein in contrast to its homologous Synechocystis ArsC protein. Expression of Alr1105 enhanced the arsenic tolerance in the arsenate reductase mutant E. coli WC3110 (∆arsC) and rendered better growth than the wild type W3110 up to 40 mM As (V). Notwithstanding above, the recombinant E. coli strain when exposed to CdCl₂, ZnSO₄, NiCl₂, CoCl₂, CuCl₂, heat, UV-B and carbofuron showed increase in growth over the wild type and mutant E. coli transformed with the empty vector. Furthermore, an enhanced growth of the recombinant E. coli in the presence of oxidative stress producing chemicals (MV, PMS and H₂O₂), suggested its protective role against these stresses. Appreciable expression of alr1105 gene as measured by qRT-PCR at different time points under selected stresses reconfirmed its role in stress tolerance. Thus the Alr1105 of Anabaena sp. PCC7120 functions as an arsenate reductase and possess novel properties different from the arsenate reductases known so far.

  17. Ar-40-Ar-39 microanalysis of single 74220 glass balls and 72435 breccia clasts

    NASA Technical Reports Server (NTRS)

    Huneke, J. C.

    1978-01-01

    Ar-40-Ar-39 age measurements on single orange glass balls from the Apollo 17 soil 74220 and individual clasts from the Apollo 17 highland breccia 72435 are reported. The measurements required the use of newly established microanalytical techniques to obtain high quality analyses on about 0.5 mg particles with only a few hundred ppm K. An age of 3.60 plus or minus 0.04 b.y. is determined for the orange glass. No corrections for a trapped Ar-40 component were required. The glass forming event occurred at the very end of or after the extrusion of the mare basalts at the Apollo 17 site. An extremely well defined age plateau at 3.86 plus or minus 0.04 b.y. was determined for a 72435 plagioclase clast with attached matrix. A second large plagioclase crystal yielded significantly older ages over the last 60% of Ar release at high temperatures and is a relict clast incompletely degassed at the time of breccia formation. 72435 also contains plagioclase clasts with primitive Sr and a 4.55 AE old dunite clast. The Ar results provide additional evidence for the association of chemically unequilibrated, relict clasts with both primitive Sr and older K/Ar ages.

  18. Chlorate reductase is cotranscribed with cytochrome c and other downstream genes in the gene cluster for chlorate respiration of Ideonella dechloratans.

    PubMed

    Hellberg Lindqvist, Miriam; Nilsson, Thomas; Sundin, Pontus; Rova, Maria

    2015-03-01

    The chlorate-respiring bacterium Ideonella dechloratans is a facultative anaerobe that can use both oxygen and chlorate as terminal electron acceptors. The genes for the enzymes chlorate reductase (clrABDC) and chlorite dismutase, necessary for chlorate metabolism and probably acquired by lateral gene transfer, are located in a gene cluster that also includes other genes potentially important for chlorate metabolism. Among those are a gene for cytochrome c (cyc) whose gene product may serve as an electron carrier during chlorate reduction, a cofactor biosynthesis gene (mobB) and a predicted transcriptional regulator (arsR). Only chlorate reductase and chlorite dismutase have been shown to be expressed in vivo. Here, we report the in vivo production of a single polycistronic transcript covering eight open reading frames including clrABDC, cyc, mobB and arsR. Transcription levels of the cyc and clrA genes were compared to each other by the use of qRT-PCR in RNA preparations from cells grown under aerobic or chlorate reducing anaerobic conditions. The two genes showed the same mRNA levels under both growth regimes, indicating that no transcription termination occurs between them. Higher transcription levels were observed at growth without external oxygen supply. Implications for electron pathway integration following lateral gene transfer are discussed. PMID:25673284

  19. Application of the 40Ar/39Ar technique to date the Minoan Tuff, Santorini

    NASA Astrophysics Data System (ADS)

    Wijbrans, J. R.; Kuiper, K.; Morgan, L. E.; Klaver, M.; Vroon, P. Z.

    2012-12-01

    The age of the catastrophic eruption of the volcano of Santorini during the Bronze Age is well established from 14C dating at 3344.9 ± 7.5 a1 (uncertainties quoted as 1-σ). Application of the 40Ar/39Ar technique to products from this eruption is used here to (1) investigate the limits of the technique using conventional single collector mass spectrometry on a MAP215-50 instrument, (2) analyse sources of uncertainty to identify major contributing factors for the uncertainty of young 40Ar/39Ar ages, and (3) provide 40Ar/39Ar ages for a sample that has been previously dated via 14C and dendrochronology to further investigate issues with the accuracy of 40Ar/39Ar dating in the late Quaternary. We have separated the plagioclase fraction from the lower Minoan Tuff that immediately overlies the Cape Riva (rp6) tuff in a bay on the west coast of Thira, NW of the town of Oia. Using the calibration of 40Ar/36Ar of Lee et al.2, the decay constant recommended by Min at al.3, and the FCs age of Kuiper et al.4, we calculate an inverse isochron age of 3.7 ± 1.6 ka and a trapped 40Ar/36Ar intercept of 299.8 ± 1.2, slightly higher than the ratio for atmospheric argon of 298.56 ± 0.31, when all steps with ages > 50 ka are included in the regression. Enrichment in radiogenic 40Ar in the steps used for the isochron is extremely low, given the low concentration of K2O in plagioclase and the extremely young age. The stepwise heating approach proved useful because in all 5 replicate experiments unexpectedly high ages showed up at higher step temperatures, suggesting that in each separate some older contaminant was present. The plateaus of each of the replicate experiments had quite reproducible ages, however, and a pooled age was calculated for 23 out of 48 individual steps. The pooled age for the plateau was 17.6 ± 4.1 ka, which is high due to the slight component of excess 40Ar in the non-radiogenic component, as revealed from regression analysis. refs: 1SW Manning et al. (2006

  20. The new 39Ar/40Ar dating facility of the LSCE, background and performances.

    NASA Astrophysics Data System (ADS)

    Scaillet, S.; Nomade, S.; Guillou, H.; Scaillet-Vita, G.

    2007-12-01

    Precise and accurate timescales are increasingly needed in most disciplines of the Earth sciences. To contribute to this challenging task, a new 39Ar/40Ar laboratory, specifically devoted to the dating of very recent (down to 1 ka) volcanic products, has been developed at the LSCE (CEA-IPSL-UVSQ, France). In this contribution, we present the first data obtained from this new facility. The laboratory comprises a VG 5400 mass spectrometer coupled with a high sensitivity and high dynamic range ion counting system. Gas extraction is achieved with a 25 watts C02 laser or a double vacuum furnace depending of the analyzed samples. The full metal vacuum and purification line feature a GP50 and a compact Ti flash getters which permit extremely low blank for all Argon isotopes (e.g. ~ 3.0 10e-19 Moles for 36Ar). Both analytical protocols and hardware were specifically developed and optimized to date extremely young samples. Analytical performances including protocols, flux monitoring as well mass spectrometer discrimination correction method will be presented in the light of data obtained over the last 10 months. All samples were irradiated, under cadmium, in the Β-1 position (~1.0 10e+13 fast n cm-2 s-1) of the 70MWh-1 OSIRIS reactor (Pierre Süe laboratory, CEA-Saclay, France). Irradiation package is composed of home-design Aluminium disks constituting a 4 cm stack (10 to 30 unknowns/irradiation). Analyzed neutrons flux standards indicate less than 1% variation along the 4cm stack and validate the use of this reactor for high-precision 39Ar/40Ar dating. The precision and accuracy of the facility has been checked from cross-comparison of international single grain standards including FCs (28.02Ma), ACR-2 (1.194Ma) and TCR (28.32Ma) using the most recent recommended values for these monitors. A total of 80 grains in two irradiations (10 and 120 minutes) will be presented in details. Results from single-grain analyses agree within errors with those proposed by Renne et al

  1. Calibration of a Carboniferous U-Pb and Ar-Ar Standard

    NASA Astrophysics Data System (ADS)

    Shea, E. K.; Machlus, M.; Hemming, S. R.; Bowring, S. A.

    2015-12-01

    An important goal of the EARTHTIME initiative is to produce an accurate and precise sequence of geologic events, allowing the evaluation of the rates of geologic processes. Toward this end, the Ar-Ar and U-Pb communities require a set of standards that permit inter-laboratory and inter-technique comparisons. Natural zircon and sanidine samples that are extensively analyzed by multiple labs play a critical role for these comparisons, but are currently limited in number (Plesovice, R33, Temora 2 zircons; Fish Canyon, Alder Creek, Taylor Creek sanidine). Calibration between the sanidine Ar-Ar and zircon U-Pb is sparse and complexity in one or both of the systems is a general problem. Further, there is currently no Paleozoic sanidine monitor standard for Ar-Ar geochronology. The sanidine- and zircon-bearing Carboniferous Fire Clay tonstein provides potential natural Paleozoic standards for these two systems. The Fire Clay tonstein is a voluminous Carboniferous ash bed from the Appalachian basin. Exposures of the tonstein span over 300 km, making it a valuable marker bed for Appalachian geology. Here we report the results of 64 single-grain zircon U-Pb TIMS analyses and 223 single-grain sanidine Ar/Ar analyses. Although previous efforts have been plagued by xenocrystic zircons, by careful selection of only elongate crystals we were able to entirely avoid discordant analyses older than 316 Ma. Unfiltered analyses of zircons analyzed from a population of acicular crystals give a range of dates between ~315 and ~314 Ma with 2 sigma uncertainties of ~0.2 Ma. A weighted mean of these dates has an MSWD of ~4.0, suggesting geological complexity in the magma chamber or post-eruption lead loss. Sanidine ages have a range of less than 1 %, and only a single population can be distinguished with precision at the 1 Ma level for individual crystals. The ability to select crystals of both zircon and sanidine that give a narrow range of ages suggests that the Fire Clay tonstein holds

  2. A natural laboratory for 40Ar/39Ar geochronology: ICDP cores from Lake Van, Turkey

    NASA Astrophysics Data System (ADS)

    Engelhardt, Jonathan; Sudo, Masafumi; Oberhänsli, Roland

    2015-04-01

    Pore water samples from ICDP Paleovan cores indicate a limited pore water exchange within Quaternary lake sediments. The core's volcaniclastic sections bear unaltered K-rich ternary feldspar and fresh to altered glass shards of predominantly rhyolitic composition. Whereas applying the 40Ar/39Ar method on feldspars resulted in ages timing a late-stage crystallization, glass shards had the potential to date the eruption. Volcanic glass is prone to modifications such as hydrous alteration (palagonitization) and devitrification (Cerling et al., 1985). These modifications affect the glass' chemistry and challenge the application of the 40Ar/39Ar method. Gaining precise radiometric ages from two phases has the potential to strengthen a climate-stratigraphic age-model (Stockhecke et al., 2014), and to significantly increase the temporal resolution on the deposition of the lake sediments. Vice versa the core's previous age model has the ability to question the reliability of 40Ar/39Ar eruption ages derived from ternary feldspars and glass shards. Multi- and single-grain total fusion on alkali feldspars from six volcaniclastic deposits resulted in Pleistocene ages that are in good agreement with the predicted age model. Feldspar phenocrysts from three ashes in the core's youngest section yielded consistent isochron ages that are significantly older than the model's prediction. Several distinct stratigraphic and paleomagnetic time markers of similar stratigraphic positions contradict to the older radiometric dates (Stockhecke et al., 2014). Partial resorption features of inherited feldspar domains and the involvement of excess 40Ar indicate incomplete degassing of older domains. To evaluate the magmatic history of the different domains EMPA mappings of trace elements that could be interpreted as Ar diffusion couples are currently conducted. Geochronology on Paleovan cores offers unique opportunities to monitor the effect of alteration on the Ar-systematics of volcanic glass

  3. 40Ar* loss in experimentally deformed muscovite and biotite with implications for 40Ar/39Ar geochronology of naturally deformed rocks

    USGS Publications Warehouse

    Cosca, M.; Stunitz, H.; Bourgeix, A.-L.; Lee, J.P.

    2011-01-01

    The effects of deformation on radiogenic argon (40Ar*) retentivity in mica are described from high pressure experiments performed on rock samples of peraluminous granite containing euhedral muscovite and biotite. Cylindrical cores, ???15mm in length and 6.25mm in diameter, were drilled from granite collected from the South Armorican Massif in northwestern France, loaded into gold capsules, and weld-sealed in the presence of excess water. The samples were deformed at a pressure of 10kb and a temperature of 600??C over a period 29 of hours within a solid medium assembly in a Griggs-type triaxial hydraulic deformation apparatus. Overall shortening in the experiments was approximately 10%. Transmitted light and secondary and backscattered electron imaging of the deformed granite samples reveals evidence of induced defects and for significant physical grain size reduction by kinking, cracking, and grain segmentation of the micas.Infrared (IR) laser (CO2) heating of individual 1.5-2.5mm diameter grains of muscovite and biotite separated from the undeformed granite yield well-defined 40Ar/39Ar plateau ages of 311??2Ma (2??). Identical experiments on single grains separated from the experimentally deformed granite yield results indicating 40Ar* loss of 0-35% in muscovite and 2-3% 40Ar* loss in biotite. Intragrain in situ ultraviolet (UV) laser ablation 40Ar/39Ar ages (??4-10%, 1??) of deformed muscovites range from 309??13 to 264??7Ma, consistent with 0-16% 40Ar* loss relative to the undeformed muscovite. The in situ UV laser ablation 40Ar/39Ar ages of deformed biotite vary from 301 to 217Ma, consistent with up to 32% 40Ar* loss. No spatial correlation is observed between in situ 40Ar/39Ar age and position within individual grains. Using available argon diffusion data for muscovite the observed 40Ar* loss in the experimentally treated muscovite can be utilized to predict average 40Ar* diffusion dimensions. Maximum 40Ar/39Ar ages obtained by UV laser ablation overlap those

  4. Alpha gas state in 36Ar

    NASA Astrophysics Data System (ADS)

    Akimune, Hidetoshi; Gibelin, Julien; Harakeh, Muhsin; Itoh, Masatoshi; Kawabata, Takahiro; Tamii, Atsushi; Fujiwara, Mamoru; Miki, Kenjiro; Iwamoto, Chiro; Otsu, Hideaki; Oha, Shinsuke; Tanihata, Isao; Muramoto, Tomoyuki; Kadono, Chika; Kalantar, Nasser; Ando, Shun; Leblond, Sylvian; Ayyad, Yassid; Furuno, Tatsuya; Tsynyra, Miho; Baba, Tasuo; Adachi, Satoshi; Freer, Martin

    2014-09-01

    The α cluster structures in light nuclei with N = Z are expected to appear abov the threshold energy of breakup into α particles. After the proposal of an α cluster wave function with α particle condensate type, such condensate states are both theoretically and experimentally discussed extensively. Theoretically, the existence of dilute α cluster state in nuclei with mass region of A > 16, experimentally, is not confirmed for N- α cluster states in nuclei heavier than A = 16. Recently, we measured α inelastic scattering of 36Ar followed by α decay in an inverse kinematics setup. A 50 MeV/u 36Ar beam from RCNP ring cyclotron was used to bombard a 4He gas target. α particles were detected in the magnetic spectrometer LAS which was set at 0 degrees. The α cluster structures in light nuclei with N = Z are expected to appear abov the threshold energy of breakup into α particles. After the proposal of an α cluster wave function with α particle condensate type, such condensate states are both theoretically and experimentally discussed extensively. Theoretically, the existence of dilute α cluster state in nuclei with mass region of A > 16, experimentally, is not confirmed for N- α cluster states in nuclei heavier than A = 16. Recently, we measured α inelastic scattering of 36Ar followed by α decay in an inverse kinematics setup. A 50 MeV/u 36Ar beam from RCNP ring cyclotron was used to bombard a 4He gas target. α particles were detected in the magnetic spectrometer LAS which was set at 0 degrees. Taro Hirao Grant-in-Aid for Scientific Research.

  5. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375...

  6. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375...

  7. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375...

  8. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375...

  9. 21 CFR 864.7375 - Glutathione reductase assay.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7375...

  10. Domain evolution and functional diversification of sulfite reductases.

    PubMed

    Dhillon, Ashita; Goswami, Sulip; Riley, Monica; Teske, Andreas; Sogin, Mitchell

    2005-02-01

    Sulfite reductases are key enzymes of assimilatory and dissimilatory sulfur metabolism, which occur in diverse bacterial and archaeal lineages. They share a highly conserved domain "C-X5-C-n-C-X3-C" for binding siroheme and iron-sulfur clusters that facilitate electron transfer to the substrate. For each sulfite reductase cluster, the siroheme-binding domain is positioned slightly differently at the N-terminus of dsrA and dsrB, while in the assimilatory proteins the siroheme domain is located at the C-terminus. Our sequence and phylogenetic analysis of the siroheme-binding domain shows that sulfite reductase sequences diverged from a common ancestor into four separate clusters (aSir, alSir, dsr, and asrC) that are biochemically distinct; each serves a different assimilatory or dissimilatory role in sulfur metabolism. The phylogenetic distribution and functional grouping in sulfite reductase clusters (dsrA and dsrB vs. aSiR, asrC, and alSir) suggest that their functional diversification during evolution may have preceded the bacterial/archaeal divergence.

  11. The Kinetics and Inhibition of the Enzyme Methemoglobin Reductase

    ERIC Educational Resources Information Center

    Splittgerber, A. G.; And Others

    1975-01-01

    Describes an undergraduate biochemistry experiment which involves the preparation and kinetics of an oxidation-reduction enzyme system, methemoglobin reductase. A crude enzyme extract is prepared and assayed spectrophotometrically. The enzyme system obeys Michaelis-Menton kinetics with respect to both substrate and the NADH cofactor. (MLH)

  12. Thioredoxin and NADP-thioredoxin reductase from cultured carrot cells

    NASA Technical Reports Server (NTRS)

    Johnson, T. C.; Cao, R. Q.; Kung, J. E.; Buchanan, B. B.

    1987-01-01

    Dark-grown carrot (Daucus carota L.) tissue cultures were found to contain both protein components of the NADP/thioredoxin system--NADP-thioredoxin reductase and the thioredoxin characteristic of heterotrophic systems, thioredoxin h. Thioredoxin h was purified to apparent homogeneity and, like typical bacterial counterparts, was a 12-kdalton (kDa) acidic protein capable of activating chloroplast NADP-malate dehydrogenase (EC 1.1.1.82) more effectively than fructose-1,6-bisphosphatase (EC 3.1.3.11). NADP-thioredoxin reductase (EC 1.6.4.5) was partially purified and found to be an arsenite-sensitive enzyme composed of two 34-kDa subunits. Carrot NADP-thioredoxin reductase resembled more closely its counterpart from bacteria rather than animal cells in acceptor (thioredoxin) specificity. Upon greening of the cells, the content of NADP-thioredoxin-reductase activity, and, to a lesser extent, thioredoxin h decreased. The results confirm the presence of a heterotrophic-type thioredoxin system in plant cells and raise the question of its physiological function.

  13. The arsenic hyperaccumulating Pteris vittata expresses two arsenate reductases

    NASA Astrophysics Data System (ADS)

    Cesaro, Patrizia; Cattaneo, Chiara; Bona, Elisa; Berta, Graziella; Cavaletto, Maria

    2015-09-01

    Enzymatic reduction of arsenate to arsenite is the first known step in arsenate metabolism in all organisms. Although the presence of one mRNA arsenate reductase (PvACR2) has been characterized in gametophytes of P. vittata, no arsenate reductase protein has been directly observed in this arsenic hyperaccumulating fern, yet. In order to assess the possible presence of arsenate reductase in P. vittata, two recombinant proteins, ACR2-His6 and Trx-His6-S-Pv2.5-8 were prepared in Escherichia coli, purified and used to produce polyclonal antibodies. The presence of these two enzymes was evaluated by qRT-PCR, immunoblotting and direct MS analysis. Enzymatic activity was detected in crude extracts. For the first time we detected and identified two arsenate reductase proteins (PvACR2 and Pv2.5-8) in sporophytes and gametophytes of P. vittata. Despite an increase of the mRNA levels for both proteins in roots, no difference was observed at the protein level after arsenic treatment. Overall, our data demonstrate the constitutive protein expression of PvACR2 and Pv2.5-8 in P. vittata tissues and propose their specific role in the complex metabolic network of arsenic reduction.

  14. Characterization of mitochondrial thioredoxin reductase from C. elegans

    SciTech Connect

    Lacey, Brian M.; Hondal, Robert J. . E-mail: Robert.Hondal@uvm.edu

    2006-08-04

    Thioredoxin reductase catalyzes the NADPH-dependent reduction of the catalytic disulfide bond of thioredoxin. In mammals and other higher eukaryotes, thioredoxin reductases contain the rare amino acid selenocysteine at the active site. The mitochondrial enzyme from Caenorhabditis elegans, however, contains a cysteine residue in place of selenocysteine. The mitochondrial C. elegans thioredoxin reductase was cloned from an expressed sequence tag and then produced in Escherichia coli as an intein-fusion protein. The purified recombinant enzyme has a k {sub cat} of 610 min{sup -1} and a K {sub m} of 610 {mu}M using E. coli thioredoxin as substrate. The reported k {sub cat} is 25% of the k {sub cat} of the mammalian enzyme and is 43-fold higher than a cysteine mutant of mammalian thioredoxin reductase. The enzyme would reduce selenocysteine, but not hydrogen peroxide or insulin. The flanking glycine residues of the GCCG motif were mutated to serine. The mutants improved substrate binding, but decreased the catalytic rate.

  15. A detoxifying oxygen reductase in the anaerobic protozoan Entamoeba histolytica.

    PubMed

    Vicente, João B; Tran, Vy; Pinto, Liliana; Teixeira, Miguel; Singh, Upinder

    2012-09-01

    We report the characterization of a bacterial-type oxygen reductase abundant in the cytoplasm of the anaerobic protozoan parasite Entamoeba histolytica. Upon host infection, E. histolytica is confronted with various oxygen tensions in the host intestine, as well as increased reactive oxygen and nitrogen species at the site of local tissue inflammation. Resistance to oxygen-derived stress thus plays an important role in the pathogenic potential of E. histolytica. The genome of E. histolytica has four genes that encode flavodiiron proteins, which are bacterial-type oxygen or nitric oxide reductases and were likely acquired by lateral gene transfer from prokaryotes. The EhFdp1 gene has higher expression in virulent than in nonvirulent Entamoeba strains and species, hinting that the response to oxidative stress may be one correlate of virulence potential. We demonstrate that EhFdp1 is abundantly expressed in the cytoplasm of E. histolytica and that the protein levels are markedly increased (up to ~5-fold) upon oxygen exposure. Additionally, we produced fully functional recombinant EhFdp1 and demonstrated that this enzyme is a specific and robust oxygen reductase but has poor nitric oxide reductase activity. This observation represents a new mechanism of oxygen resistance in the anaerobic protozoan pathogen E. histolytica.

  16. The arsenic hyperaccumulating Pteris vittata expresses two arsenate reductases

    PubMed Central

    Cesaro, Patrizia; Cattaneo, Chiara; Bona, Elisa; Berta, Graziella; Cavaletto, Maria

    2015-01-01

    Enzymatic reduction of arsenate to arsenite is the first known step in arsenate metabolism in all organisms. Although the presence of one mRNA arsenate reductase (PvACR2) has been characterized in gametophytes of P. vittata, no arsenate reductase protein has been directly observed in this arsenic hyperaccumulating fern, yet. In order to assess the possible presence of arsenate reductase in P. vittata, two recombinant proteins, ACR2-His6 and Trx-His6-S-Pv2.5–8 were prepared in Escherichia coli, purified and used to produce polyclonal antibodies. The presence of these two enzymes was evaluated by qRT-PCR, immunoblotting and direct MS analysis. Enzymatic activity was detected in crude extracts. For the first time we detected and identified two arsenate reductase proteins (PvACR2 and Pv2.5–8) in sporophytes and gametophytes of P. vittata. Despite an increase of the mRNA levels for both proteins in roots, no difference was observed at the protein level after arsenic treatment. Overall, our data demonstrate the constitutive protein expression of PvACR2 and Pv2.5–8 in P. vittata tissues and propose their specific role in the complex metabolic network of arsenic reduction. PMID:26412036

  17. Domain Evolution and Functional Diversification of Sulfite Reductases

    NASA Astrophysics Data System (ADS)

    Dhillon, Ashita; Goswami, Sulip; Riley, Monica; Teske, Andreas; Sogin, Mitchell

    2005-02-01

    Sulfite reductases are key enzymes of assimilatory and dissimilatory sulfur metabolism, which occur in diverse bacterial and archaeal lineages. They share a highly conserved domain "C-X5-C-n-C-X3-C" for binding siroheme and iron-sulfur clusters that facilitate electron transfer to the substrate. For each sulfite reductase cluster, the siroheme-binding domain is positioned slightly differently at the N-terminus of dsrA and dsrB, while in the assimilatory proteins the siroheme domain is located at the C-terminus. Our sequence and phylogenetic analysis of the siroheme-binding domain shows that sulfite reductase sequences diverged from a common ancestor into four separate clusters (aSir, alSir, dsr, and asrC) that are biochemically distinct; each serves a different assimilatory or dissimilatory role in sulfur metabolism. The phylogenetic distribution and functional grouping in sulfite reductase clusters (dsrA and dsrB vs. aSiR, asrC, and alSir) suggest that their functional diversification during evolution may have preceded the bacterial/archaeal divergence.

  18. Molecular genetics of steroid 5 alpha-reductase 2 deficiency.

    PubMed Central

    Thigpen, A E; Davis, D L; Milatovich, A; Mendonca, B B; Imperato-McGinley, J; Griffin, J E; Francke, U; Wilson, J D; Russell, D W

    1992-01-01

    Two isozymes of steroid 5 alpha-reductase encoded by separate loci catalyze the conversion of testosterone to dihydrotestosterone. Inherited defects in the type 2 isozyme lead to male pseudohermaphroditism in which affected males have a normal internal urogenital tract but external genitalia resembling those of a female. The 5 alpha-reductase type 2 gene (gene symbol SRD5A2) was cloned and shown to contain five exons and four introns. The gene was localized to chromosome 2 band p23 by somatic cell hybrid mapping and chromosomal in situ hybridization. Molecular analysis of the SRD5A2 gene resulted in the identification of 18 mutations in 11 homozygotes, 6 compound heterozygotes, and 4 inferred compound heterozygotes from 23 families with 5 alpha-reductase deficiency. 6 apparent recurrent mutations were detected in 19 different ethnic backgrounds. In two patients, the catalytic efficiency of the mutant enzymes correlated with the severity of the disease. The high proportion of compound heterozygotes suggests that the carrier frequency of mutations in the 5 alpha-reductase type 2 gene may be higher than previously thought. Images PMID:1522235

  19. 5. cap alpha. -reductase activity in rat adipose tissue

    SciTech Connect

    Zyirek, M.; Flood, C.; Longcope, C.

    1987-11-01

    We measured the 5 ..cap alpha..-reductase activity in isolated cell preparations of rat adipose tissue using the formation of (/sup 3/H) dihydrotestosterone from (/sup 3/H) testosterone as an endpoint. Stromal cells were prepared from the epididymal fat pad, perinephric fat, and subcutaneous fat of male rats and from perinephric fat of female rats. Adipocytes were prepared from the epididymal fat pad and perinephric fat of male rats. Stromal cells from the epididymal fat pad and perinephric fat contained greater 5..cap alpha..-reductase activity than did the adipocytes from these depots. Stromal cells from the epididymal fat pad contained greater activity than those from perinephric and subcutaneous depots. Perinephric stromal cells from female rats were slightly more active than those from male rats. Estradiol (10/sup -8/ M), when added to the medium, caused a 90% decrease in 5..cap alpha..-reductase activity. Aromatase activity was minimal, several orders of magnitude less than 5..cap alpha..-reductase activity in each tissue studied.

  20. ARSENICALS INHIBIT THIOREDOXIN REDUCTASE ACTIVITY IN CULTURED RAT HEPATOCYTES

    EPA Science Inventory

    ARSENICALS INHIBIT THIOREDOXIN REDUCTASE ACTIVITY IN CULTURED RAT HEPATOCYTES.

    S. Lin1, L. M. Del Razo1, M. Styblo1, C. Wang2, W. R. Cullen2, and D.J. Thomas3. 1Univ. North Carolina, Chapel Hill, NC; 2Univ. British Columbia, Vancouver, BC, Canada; 3National Health and En...

  1. Comparison of Ca and Ar Diffusion in Phlogopite: Implications for K-Ca and K-Ar Geochronology

    NASA Astrophysics Data System (ADS)

    Cruz, M. F.; Szilas, K.; Grove, M. J.

    2015-12-01

    Coupled geochronology based upon branched decay of 40K-40Ar and 40K-40Ca decay is rarely exploited because 40Ca is the major common isotope of calcium and 40Ca and 40K are difficult to resolve isotopically without resorting to isotope dilution wet chemistry. Recently developed ion microprobe methods based upon measurement of doubly ionized species partially overcome the latter problem and have been applied to high K/Ca micas. The ability to interpret K-Ar and K-Ca results is limited due to uncertainty in the relative diffusion properties of Ca and Ar. To address this problem, we are performing Ar and Ca diffusion experiments and fluid-crystal Ar partitioning experiments with anhydrous F-phlogopite that is stable to 1390°C. As an additional check, we are comparing K-Ca and K-Ar ages from natural mantle phlogopites from a variety of settings to assess the relative retentivity of Ar and Ca. South African xenoliths tend to yield 40Ar/39Ar ages that are much older than K-Ca ages from the same phologopites. Possible excess 40Ar and high common Ca render the comparisons inconclusive, but this suggests that Ca diffuses more readily than Ar in phlogopite. Our most definitive K-Ca phlogopite results (i.e., least affected by common Ca) come from the Archean Seqi dunite of SW Greenland. The K-Ca ages of Seqi phlogopites is 927 ± 26 Ma (2s). Incremental heating 40Ar/39Ar results from the same sample yields a much older result with a terminal age of 3.5 Ga. However, the first 5-10% of 39Ar release are consistent with transient heating at ca. 900 Ma. Considered together, the K-Ca and 40Ar/39Ar results from the Seqi dunite locality strongly suggest that Ca diffusion is more rapid than Ar diffusion in phlogopite.

  2. Precise K-Ar, 40Ar/39Ar, Rb-Sr and U/Pb mineral ages from the 27.5 Ma fish canyon tuff reference standard

    USGS Publications Warehouse

    Lanphere, M.A.; Baadsgaard, H.

    2001-01-01

    The accuracy of ages measured using the 40Ar/39Ar technique is affected by uncertainties in the age of radiation fluence-monitor minerals. At present, there is lack of agreement about the ages of certain minerals used as fluence monitors. The accuracy of the age of a standard may be improved if the age can be measured using different decay schemes. This has been done by measuring ages on minerals from the Oligocene Fish Canyon Tuff (FCT) using the K-Ar, 40Ar/39Ar. Rb-Sr and U/Pb methods. K-Ar and 40Ar/39Ar total fusion ages of sanidine, biotite and hornblende yielded a mean age of 27.57 ?? 0.36 Ma. The weighted mean 40Ar/39Ar plateau age of sanidine and biotite is 27.57 ?? 0.18 Ma. A biotite-feldspar Rb-Sr isochron yielded an age of 27.44 ?? 0.16 Ma. The U-Pb data for zircon are complex because of the presence of Precambrian zircons and inheritance of radiogenic Pb. Zircons with 207Pb/235U < 0.4 yielded a discordia line with a lower concordia intercept of 27.52 ?? 0.09 Ma. Evaluation of the combined data suggests that the best age for FCT is 27.51 Ma. Published by Elsevier Science B.V.

  3. Differential molecular response of monodehydroascorbate reductase and glutathione reductase by nitration and S-nitrosylation

    PubMed Central

    Begara-Morales, Juan C.; Sánchez-Calvo, Beatriz; Chaki, Mounira; Mata-Pérez, Capilla; Valderrama, Raquel; Padilla, María N.; Luque, Francisco; Corpas, Francisco J.; Barroso, Juan B.

    2015-01-01

    The ascorbate–glutathione cycle is a metabolic pathway that detoxifies hydrogen peroxide and involves enzymatic and non-enzymatic antioxidants. Proteomic studies have shown that some enzymes in this cycle such as ascorbate peroxidase (APX), monodehydroascorbate reductase (MDAR), and glutathione reductase (GR) are potential targets for post-translational modifications (PMTs) mediated by nitric oxide-derived molecules. Using purified recombinant pea peroxisomal MDAR and cytosolic and chloroplastic GR enzymes produced in Escherichia coli, the effects of peroxynitrite (ONOO–) and S-nitrosoglutathione (GSNO) which are known to mediate protein nitration and S-nitrosylation processes, respectively, were analysed. Although ONOO– and GSNO inhibit peroxisomal MDAR activity, chloroplastic and cytosolic GR were not affected by these molecules. Mass spectrometric analysis of the nitrated MDAR revealed that Tyr213, Try292, and Tyr345 were exclusively nitrated to 3-nitrotyrosine by ONOO–. The location of these residues in the structure of pea peroxisomal MDAR reveals that Tyr345 is found at 3.3 Å of His313 which is involved in the NADP-binding site. Site-directed mutagenesis confirmed Tyr345 as the primary site of nitration responsible for the inhibition of MDAR activity by ONOO–. These results provide new insights into the molecular regulation of MDAR which is deactivated by nitration and S-nitrosylation. However, GR was not affected by ONOO– or GSNO, suggesting the existence of a mechanism to conserve redox status by maintaining the level of reduced GSH. Under a nitro-oxidative stress induced by salinity (150mM NaCl), MDAR expression (mRNA, protein, and enzyme activity levels) was increased, probably to compensate the inhibitory effects of S-nitrosylation and nitration on the enzyme. The present data show the modulation of the antioxidative response of key enzymes in the ascorbate–glutathione cycle by nitric oxide (NO)-PTMs, thus indicating the close involvement

  4. Measurement of nitrous oxide reductase activity in aquatic sediments

    USGS Publications Warehouse

    Miller, L.G.; Oremland, R.S.; Paulsen, S.

    1986-01-01

    Denitrification in aquatic sediments was measured by an N2O reductase assay. Sediments consumed small added quantities of N2O over short periods (a few hours). In experiments with sediment slurries, N2O reductase activity was inhibited by O2, C2H2, heat treatment, and by high levels of nitrate (1 mM) or sulfide (10 mM). However, ambient levels of nitrate (<100 μM) did not influence activity, and moderate levels (about 150 μM) induced only a short lag before reductase activity began. Moderate levels of sulfide (<1 mM) had no effect on N2O reductase activity. Nitrous oxide reductase displayed Michaelis-Menten kinetics in sediments from freshwater (Km = 2.17 μM), estuarine (Km = 14.5 μM), and alkaline-saline (Km = 501 μM) environments. An in situ assay was devised in which a solution of N2O was injected into sealed glass cores containing intact sediment. Two estimates of net rates of denitrification in San Francisco Bay under approximated in situ conditions were 0.009 and 0.041 mmol of N2O per m2 per h. Addition of chlorate to inhibit denitrification in these intact-core experiments (to estimate gross rates of N2O consumption) resulted in approximately a 14% upward revision of estimates of net rates. These results were comparable to an in situ estimate of 0.022 mmol of N2O per m2 per h made with the acetylene block assay.

  5. 3-Oxoacyl-[ACP] reductase from oilseed rape (Brassica napus).

    PubMed

    Sheldon, P S; Kekwick, R G; Smith, C G; Sidebottom, C; Slabas, A R

    1992-04-01

    3-Oxoacyl-[ACP] reductase (E.C. 1.1.1.100, alternatively known as beta-ketoacyl-[ACP] reductase), a component of fatty acid synthetase has been purified from seeds of rape by ammonium sulphate fractionation, Procion Red H-E3B chromatography, FPLC gel filtration and high performance hydroxyapatite chromatography. The purified enzyme appears on SDS-PAGE as a number of 20-30 kDa components and has a strong tendency to exist in a dimeric form, particularly when dithiothreitol is not present to reduce disulphide bonds. Cleveland mapping and cross-reactivity with antiserum raised against avocado 3-oxoacyl-[ACP] reductase both indicate that the multiple components have similar primary structures. On gel filtration the enzyme appears to have a molecular mass of 120 kDa suggesting that the native structure is tetrameric. The enzyme has a strong preference for the acetoacetyl ester of acyl carrier protein (Km = 3 microM) over the corresponding esters of the model substrates N-acetyl cysteamine (Km = 35 mM) and CoA (Km = 261 microM). It is inactivated by dilution but this can be partly prevented by the inclusion of NADPH. Using an antiserum prepared against avocado 3-oxoacyl-[ACP] reductase, the enzyme has been visualised inside the plastids of rape embryo and leaf tissues by immunoelectron microscopy. Amino acid sequencing of two peptides prepared by digestion of the purified enzyme with trypsin showed strong similarities with 3-oxoacyl-[ACP] reductase from avocado pear and the Nod G gene product from Rhizobium meliloti.

  6. Integration of Mobile AR Technology in Performance Assessment

    ERIC Educational Resources Information Center

    Kuo-Hung, Chao; Kuo-En, Chang; Chung-Hsien, Lan; Kinshuk; Yao-Ting, Sung

    2016-01-01

    This study was aimed at exploring how to use augmented reality (AR) technology to enhance the effect of performance assessment (PA). A mobile AR performance assessment system (MARPAS) was developed by integrating AR technology to reduce the limitations in observation and assessment during PA. This system includes three modules: Authentication, AR…

  7. Antarlides: A New Type of Androgen Receptor (AR) Antagonist that Overcomes Resistance to AR-Targeted Therapy.

    PubMed

    Saito, Shun; Fujimaki, Takahiro; Panbangred, Watanalai; Igarashi, Yasuhiro; Imoto, Masaya

    2016-02-18

    Prostate cancer is treated with androgen receptor (AR) antagonists but most patients experience disease progression after long-term treatment with these compounds. Therefore, new AR antagonists are required for patient follow-up treatment. In the course of screening for a new AR antagonist, we isolated the novel compounds antarlides A-E (1-5) from Streptomyces sp. BB47. Antarlides are mutually isomeric with respect to the double bond and have a 22-membered-ring macrocyclic structure. The full stereostructure of 1 was established by chemical modifications, including methanolysis, the Trost method, acetonide formation, and the PGME method. 1-5 inhibited the binding of androgen to ARs in vitro. In addition, 2 inhibited the transcriptional activity of not only wild-type AR but also mutant ARs, which are seen in patients with acquired resistance to clinically used AR antagonists. Therefore, antarlides are a potent new generation of AR antagonists that overcome resistance.

  8. Tracking fluid action in Little Sark Pluton (Channel Islands, UK): a UV laserprobe Ar/Ar study of excess 40Ar

    NASA Astrophysics Data System (ADS)

    Schwenzer, S. P.; Schwanethal, J.; Tindle, A. G.; Kelley, S. P.; Sherlock, S. C.

    2011-12-01

    Little Sark pluton (Channel Islands, UK) is a weakly to moderately deformed pluton of quartz diorite composition. Zircon from Little Sark has been dated as 611.4(+2.1/-1.3) Ma [Miller et al. 1991, Tectonophysics, 312], whereas titanite has an age of 606.2±0.6 Ma. Hornblende yields an Ar-Ar age of 606.4±3.4 Ma [Dallmeyer et al., 1991, J. Geol. Soc. London 148]. The rocks show evidence for post-magmatic hydrous alteration including chloritisation of biotite, and sericite formation in feldspar. We investigated the alteration in order to understand its influence on the Ar-Ar system. Detailed petrographic investigations of two samples indicate different deformation levels - S1, (less deformed), and S2 (more deformed). The feldspar is andesine to oligoclase in composition, but contains K-rich areas with patchy and sometimes cleavage guided sericitization. Mafic minerals include hornblende and biotite (the latter partially chloritized). Ar-Ar investigations were carried out using high spatial resolution UV laser ablation gas release in combination with a Nu instruments Noblesse gas mass spectrometer using spots of 80 to 140 μm diameter. Mafic minerals in S1 have highly variable 38Ar/39Ar and 37Ar/39Ar ratios and unusually high atmospheric contents for such old samples, possibly reflecting the presence of fluid inclusions associated with the sericite, particularly in the feldspar. The apparent ages in their entirety range from 429 to 668 Ma. Hornblende yields homogeneous apparent ages, whereas the younger and older ages are measured in both micas and feldspars, whose apparent ages range from 245-670 Ma. The modal age for feldspar is around 400-450 Ma, but this may not have geological meaning. Sample S2 shows many of the same features as S1. Mafic minerals span a slightly smaller age range and feldspars display apparent ages of ~245-645 Ma with a mode around 480 Ma. The alteration of both samples appears to have been accompanied by the introduction of 40Ar-rich fluids

  9. Identification and characterization of 2-naphthoyl-coenzyme A reductase, the prototype of a novel class of dearomatizing reductases.

    PubMed

    Eberlein, Christian; Estelmann, Sebastian; Seifert, Jana; von Bergen, Martin; Müller, Michael; Meckenstock, Rainer U; Boll, Matthias

    2013-06-01

    The enzymatic dearomatization of aromatic ring systems by reduction represents a highly challenging redox reaction in biology and plays a key role in the degradation of aromatic compounds under anoxic conditions. In anaerobic bacteria, most monocyclic aromatic growth substrates are converted to benzoyl-coenzyme A (CoA), which is then dearomatized to a conjugated dienoyl-CoA by ATP-dependent or -independent benzoyl-CoA reductases. It was unresolved whether or not related enzymes are involved in the anaerobic degradation of environmentally relevant polycyclic aromatic hydrocarbons (PAHs). In this work, a previously unknown dearomatizing 2-naphthoyl-CoA reductase was purified from extracts of the naphthalene-degrading, sulphidogenic enrichment culture N47. The oxygen-tolerant enzyme dearomatized the non-activated ring of 2-naphthoyl-CoA by a four-electron reduction to 5,6,7,8-tetrahydro-2-naphthoyl-CoA. The dimeric 150 kDa enzyme complex was composed of a 72 kDa subunit showing sequence similarity to members of the flavin-containing 'old yellow enzyme' family. NCR contained FAD, FMN, and an iron-sulphur cluster as cofactors. Extracts of Escherichia coli expressing the encoding gene catalysed 2-naphthoyl-CoA reduction. The identified NCR is a prototypical enzyme of a previously unknown class of dearomatizing arylcarboxyl-CoA reductases that are involved in anaerobic PAH degradation; it fundamentally differs from known benzoyl-CoA reductases.

  10. Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases

    NASA Technical Reports Server (NTRS)

    Min, Tongpil; Kasahara, Hiroyuki; Bedgar, Diana L.; Youn, Buhyun; Lawrence, Paulraj K.; Gang, David R.; Halls, Steven C.; Park, HaJeung; Hilsenbeck, Jacqueline L.; Davin, Laurence B.; Lewis, Norman G.; Kang, ChulHee

    2003-01-01

    Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

  11. Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases.

    PubMed

    Min, Tongpil; Kasahara, Hiroyuki; Bedgar, Diana L; Youn, Buhyun; Lawrence, Paulraj K; Gang, David R; Halls, Steven C; Park, HaJeung; Hilsenbeck, Jacqueline L; Davin, Laurence B; Lewis, Norman G; Kang, ChulHee

    2003-12-12

    Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

  12. (+)-Pinoresinol/(+)-lariciresinol reductase from Forsythia intermedia. Protein purification, cDNA cloning, heterologous expression and comparison to isoflavone reductase.

    PubMed

    Dinkova-Kostova, A T; Gang, D R; Davin, L B; Bedgar, D L; Chu, A; Lewis, N G

    1996-11-15

    Lignans are a widely distributed class of natural products, whose functions and distribution suggest that they are one of the earliest forms of defense to have evolved in vascular plants; some, such as podophyllotoxin and enterodiol, have important roles in cancer chemotherapy and prevention, respectively. Entry into lignan enzymology has been gained by the approximately 3000-fold purification of two isoforms of (+)-pinoresinol/(+)-lariciresinol reductase, a pivotal branchpoint enzyme in lignan biosynthesis. Both have comparable ( approximately 34.9 kDa) molecular mass and kinetic (Vmax/Km) properties and catalyze sequential, NADPH-dependent, stereospecific, hydride transfers where the incoming hydride takes up the pro-R position. The gene encoding (+)-pinoresinol/(+)-lariciresinol reductase has been cloned and the recombinant protein heterologously expressed as a functional beta-galactosidase fusion protein. Its amino acid sequence reveals a strong homology to isoflavone reductase, a key branchpoint enzyme in isoflavonoid metabolism and primarily found in the Fabaceae (angiosperms). This is of great evolutionary significance since both lignans and isoflavonoids have comparable plant defense properties, as well as similar roles as phytoestrogens. Given that lignans are widespread from primitive plants onwards, whereas the isoflavone reductase-derived isoflavonoids are mainly restricted to the Fabaceae, it is tempting to speculate that this branch of the isoflavonoid pathway arose via evolutionary divergence from that giving the lignans.

  13. Ar-39-Ar-40 Age Dating Of Two Angrites and Two Brachinites

    NASA Technical Reports Server (NTRS)

    Garrison, Daniel; Bogard, Donald

    2003-01-01

    Angrites are a rare group (approximately 7 known) of igneous meteorites with basalt-like composition, which probably derive from a relatively small parent body that differs from those of other igneous meteorites. Angrites show evidence for extinct Mn-53, Sm-146, and Pu-244, and precise U-Pb, and Pb-Pb ages of 4.558 Gyr for two angrites define the time of early parent body differentiation. The Sm-147-Nd-143 ages of two angrites range between 4.53 +/- 0.04 and 4.56 +/- 0.04 Gyr, but no Ar-39-Ar-40 or Rb-Sr ages have been reported. Most angrites show no evidence for either shock brecciation or metamorphism. Brachinites are another very rare group' of differentiated meteorites consisting primarily of olivine, with minor augite, chromite, Fe-sulfides, and sometimes plagioclase and opx. Presence of excess Xe-129 and excess Cr53 from decay of Mn-53 in some brachinites indicate that they also formed very early. Brachinite petrogenesis is poorly defined. They may be igneous cumulates or metamorphic products of chondritic-like starting material. If after their formation, angrites and brachinites cooled quickly with minimal subsequent heating, then one might expect them to show uniquely old K-Ar ages, at least in comparison to other differentiated meteorites such as eucrites and mesosiderites. Most angrites and brachinites contain very little, if any K-feldspar, which has deterred measurements of their Ar-Ar ages. We made Ar-39-Ar-40 analyses on two angrites, LEW86010 (metamorphosed) and D'Orbigny, and on two brachinites, EET99402 and Brachina. All are finds. Any feldspar in angrites is highly calcic, with expected K concentrations of <100 ppm. We selected LEW86010 and D'Orbigny because they have been the objects of several other studies and because chemical analyses suggested [K] was approximately 70 ppm in both meteorites. Brachina contains approximately 9.9% plagioclase of higher K-content than angrites, and EET99402 is estimated to contain approximately 5% K

  14. Potassium isotopic compositions of NIST potassium standards and 40Ar/39Ar mineral standards

    NASA Astrophysics Data System (ADS)

    Morgan, L. E.; Tappa, M.; Ellam, R. M.; Mark, D. F.; Lloyd, N. S.; Higgins, J. A.; Simon, J. I.

    2013-12-01

    Knowledge of the isotopic ratios of standards, spikes, and reference materials is fundamental to the accuracy of many geochronological methods. For example, the 238U/235U ratio relevant to U-Pb geochronology was recently re-determined [1] and shown to differ significantly from the previously accepted value employed during age determinations. These underlying values are fundamental to accurate age calculations in many isotopic systems, and uncertainty in these values can represent a significant (and often unrecognized) portion of the uncertainty budget for determined ages. The potassium isotopic composition of mineral standards, or neutron flux monitors, is a critical, but often overlooked component in the calculation of K-Ar and 40Ar/39Ar ages. It is currently assumed that all terrestrial materials have abundances indistinguishable from that of NIST SRM 985 [2]; this is apparently a reasonable assumption at the 0.25‰ level (1σ) [3]. The 40Ar/39Ar method further relies on the assumption that standards and samples (including primary and secondary standards) have indistinguishable 40K/39K values. We will present data establishing the potassium isotopic compositions of NIST isotopic K SRM 985, elemental K SRM 999b, and 40Ar/39Ar biotite mineral standard GA1550 (sample MD-2). Stable isotopic compositions (41K/39K) were measured by the peak shoulder method with high resolution MC-ICP-MS (Thermo Scientific NEPTUNE Plus), using the accepted value of NIST isotopic SRM 985 [2] for fractionation [4] corrections [5]. 40K abundances were measured by TIMS (Thermo Scientific TRITON), using 41K/39K values from ICP-MS measurements (or, for SRM 985, values from [2]) for internal fractionation corrections. Collectively these data represent an important step towards a metrologically traceable calibration of 40K concentrations in primary 40Ar/39Ar mineral standards and improve uncertainties by ca. an order of magnitude in the potassium isotopic compositions of standards. [1] Hiess

  15. Call for Development of New Mineral Standards for 40Ar/39Ar Dating

    NASA Astrophysics Data System (ADS)

    Deino, A. L.; Turrin, B. D.; Renne, P. R.; Hemming, S. R.

    2015-12-01

    Age determination via the 40Ar/39Ar dating method relies on the intercomparison of measured 40Ar*/39ArK ratios of geological unknowns with those of co-irradiated mineral standards. Good analytical procedure dictates that these ratios (and the evolution of the Ar ion beams underpinning them) be as similar as practical for the greatest accuracy. Unfortunately, throughout several intervals of the geological time scale this 'best practice' cannot be achieved with existing widely used standards. Only two internationally utilized sanidine standards are available for the middle to late Cenozoic: the Alder Creek Rhyolite sanidine (ACs), at ~1.2 Ma (Turrin et al., 1994; Nomade et al., 2005), and the Fish Canyon Tuff sanidine (FCs) at ~28.2 Ma (e.g., Kuiper et al., 2008; Renne et al, 2011). The situation is even worse throughout much of the rest of the Phanerozoic, as the next oldest standard in common use is the Hb3gr hornblende standard with an age of ~1.1 Ga (Turner, 1971; Jourdan et al., 2006). We propose, as a community effort, the development a set of standards covering the entire target range of high-precision 40Ar/39Ar dating, i.e. the Phanerozoic. Their ages would be stepped in a regular fashion with no more than approximately a factor of 3 between standards, such that in the worse case the 40Ar*/39Ar ratios of standards and unknown need differ by no more than a factor of two. While somewhat arbitrary, an approximately 3 X age progression allows the entire time scale to be covered by a manageable number of standards. Anchoring the progression in the widely used ACs, FCs, and Hb3gr (in bold, below) yields the following set of suggested standard ages: 0.4, 1.2, 3.3, 9.4, 28.2, 95, 320, and 1100 Ma. A suitable standard should be highly reproducible in age at the grain-to-grain and sub-grain levels, and highly radiogenic. The mineral should be abundant and easily separated from the host rock. These criteria may be most easily achieved by focusing on sanidine phenocrysts

  16. A test of the 40Ar/39Ar age spectrum technique on some terrestrial materials

    USGS Publications Warehouse

    Lanphere, M.A.; Brent, Dalrymple G.

    1971-01-01

    40Ar/39Ar age spectra were determined for 10 terrestrial rock and mineral samples whose geologic history is known from independent evidence. The spectra for six mineral and whole rock samples, including biotite, feldspar, hornblende, muscovite, and granodiorite, that have experienced post-crystallization heating did not reveal the age of crystallization in any obvious way. Minima in the spectra, however, give reasonable maximum ages for reheating and high-temperature maxima can be interpreted as minimum crystallization ages. High-temperature ages of microcline and albite that have not been reheated are approximately 10% younger than the known crystallization age. Apparently there are no domains in these feldspars that have retained radiogenic 40Ar quantitatively. Spectra from two diabase samples that contain significant quantities of excess argon might mistakenly be interpreted as spectra from reheated samples and do not give the age of emplacement. The 40Ar/39Ar age spectrum technique may be a potentially valuable tool for the study of geologic areas with complex histories, but the interpretation of age spectra from terrestrial samples seems to be more difficult than suggested by some previous studies. ?? 1971.

  17. Cosmic Ray Exposure Ages, Ar-Ar Ages, and the Origin and History of Eucrites

    NASA Technical Reports Server (NTRS)

    Wakefield, Kelli; Bogard, Donald; Garrison, Daniel

    2004-01-01

    HED meteorites likely formed at different depths on the large asteroid 4-Vesta, but passed through Vesta-derived, km-sized intermediary bodies (Vestoids), before arriving at Earth. Most eucrites and diogenites (and all howardites) are brecciated, and impact heating disturbed or reset the K-Ar ages (and some Rb-Sr ages) of most eucrites in the time period of approx. 3.4 - 4.1 Gyr ago. Some basaltic eucrites and most cumulate eucrites, however, are not brecciated. We recently showed that the Ar-39 - Ar-40 ages for several of these eucrites tightly cluster about a value of 4.48 +/- 0.02 Gyr, and we argue that this time likely represents a single large impact event on Vesta, which ejected these objects from depth and quenched their temperatures. A different parent body has been suggested for cumulate eucrites, although the Ar-Ar ages argue for a common parent. Similarities in the cosmic-ray (space) exposure ages for basaltic eucrites and diogenites also have been used to infer a common parent body for some HEDs. Here we present CRE ages of several cumulate and unbrecciated basaltic (UB) eucrites and compare these with CRE ages of other HEDs. This comparison also has some interesting implications for the relative locations of various HED types on Vesta and/or the Vestoids.

  18. First 40Ar/39Ar dating of intense Late Palaeogene lateritic weathering in Peninsular India

    NASA Astrophysics Data System (ADS)

    Bonnet, Nicolas J.; Beauvais, Anicet; Arnaud, Nicolas; Chardon, Dominique; Jayananda, Mudlappa

    2014-01-01

    Lateritic surface processes have shaped large platform and cratons of the tropical belt. Constraining the timing of such processes is crucial to decipher their role in cratonic morphogenesis and their response to long-term climatic change and lithospheric deformation. Weathering histories have been documented for South America, Africa and Australia, but precise time constraints of the lateritic weathering processes in South India are still lacking. We present 40Ar/39Ar ages of supergene cryptomelane (K-Mn oxide) formed in the Sandur Mn ore deposits exposed on the highest lateritic paleolandsurface that once covered the Mysore plateau and the adjacent Deccan Traps. Significant 40Ar/39Ar ages are estimated between ∼36 and ∼26 Ma from well-defined plateaus in step heating 39Ar release spectra and from best-fitted inverse isochrones. These ages constitute firm time constraints that document intense late Eocene-Oligocene lateritic weathering over Peninsular India under the influence of warm and wet climate comparable to that prevailing in tropical humid forests. These results imply that Southern India was weathered between ∼36 and 26 Ma and may have been dissected mostly in the Neogene.

  19. Interaction of Thermus thermophilus, ArsC enzyme and gold nanoparticles naked-eye assays speciation between As(III) and As(V)

    NASA Astrophysics Data System (ADS)

    Politi, Jane; Spadavecchia, Jolanda; Fiorentino, Gabriella; Antonucci, Immacolata; Casale, Sandra; De Stefano, Luca

    2015-10-01

    The thermophilic bacterium Thermus thermophilus HB27 encodes chromosomal arsenate reductase (TtArsC), the enzyme responsible for resistance to the harmful effects of arsenic. We report on adsorption of TtArsC onto gold nanoparticles for naked-eye monitoring of biomolecular interaction between the enzyme and arsenic species. Synthesis of hybrid biological-metallic nanoparticles has been characterized by transmission electron microscopy (TEM), ultraviolet-visible (UV-vis), dynamic light scattering (DLS) and phase modulated infrared reflection absorption (PM-IRRAS) spectroscopies. Molecular interactions have been monitored by UV-vis and Fourier transform-surface plasmon resonance (FT-SPR). Due to the nanoparticles’ aggregation on exposure to metal salts, pentavalent and trivalent arsenic solutions can be clearly distinguished by naked-eye assay, even at 85 μM concentration. Moreover, the assay shows partial selectivity against other heavy metals.

  20. Characterization of recombinant glutathione reductase from the psychrophilic Antarctic bacterium Colwellia psychrerythraea.

    PubMed

    Ji, Mikyoung; Barnwell, Callie V; Grunden, Amy M

    2015-07-01

    Glutathione reductases catalyze the reduction of oxidized glutathione (glutathione disulfide, GSSG) using NADPH as the substrate to produce reduced glutathione (GSH), which is an important antioxidant molecule that helps maintain the proper reducing environment of the cell. A recombinant form of glutathione reductase from Colwellia psychrerythraea, a marine psychrophilic bacterium, has been biochemically characterized to determine its molecular and enzymatic properties. C. psychrerythraea glutathione reductase was shown to be a homodimer with a molecular weight of 48.7 kDa using SDS-PAGE, MALDI-TOF mass spectrometry and gel filtration. The C. psychrerythraea glutathione reductase sequence shows significant homology to that of Escherichia coli glutathione reductase (66 % identity), and it possesses the FAD and NADPH binding motifs, as well as absorption spectrum features which are characteristic of flavoenzymes such as glutathione reductase. The psychrophilic C. psychrerythraea glutathione reductase exhibits higher k cat and k cat/K m at lower temperatures (4 °C) compared to mesophilic Baker's yeast glutathione reductase. However, C. psychrerythraea glutathione reductase was able to complement an E. coli glutathione reductase deletion strain in oxidative stress growth assays, demonstrating the functionality of C. psychrerythraea glutathione reductase over a broad temperature range, which suggests its potential utility as an antioxidant enzyme in heterologous systems. PMID:26101017

  1. Microwave spectroscopic and atoms in molecules theoretical investigations on the Ar···propargyl alcohol complex: Ar···H-O, Ar···π, and Ar···C interactions.

    PubMed

    Mani, Devendra; Arunan, Elangannan

    2013-03-18

    The structure of the Ar···propargyl alcohol (Ar···PA) complex is determined from the rotational spectra of the parent complex and its two deuterated isotopologues, namely Ar···PA-D(OD) and Ar···PA-D(CD). The spectra confirm a geometry in which PA exists in the gauche form with Ar located in between -OH and -C≡C-H groups. All a, b and c types of transitions show small splitting due to some large-amplitude motion dominated by C-OH torsion, as in the monomer. Splittings in a- and b-type transitions are of the order of a few kilohertz, whereas splitting in the c-type transitions is relatively larger (0.9-2.6 MHz) and decreases in the order Ar···PA>Ar···PA-D(CD)>Ar···PA-D(OD). The assignments are well supported by ab initio calculations. Atoms in molecules (AIM) and electrostatic potential calculations are used to explore the nature of the interactions in this complex. AIM calculations not only reveal the expected O-H···Ar and π···Ar interactions in the Ar···gauche-PA complex, but also novel C···Ar (of CH2OH group) and O-H···Ar interactions in the Ar···trans-PA complex. Similar interactions are also present in the Ar···methanol complex.

  2. Measuring 36Ar without H35Cl interference

    NASA Astrophysics Data System (ADS)

    Saxton, John

    2015-04-01

    Noble gas measurements are usually made in static mode, when the mass spectrometer sensitivity is inversely proportional to volume: this makes the building of very large instruments to obtain high mass resolution impracticable. A particularly challenging interference has hitherto been H35Cl, which differs in mass from 36Ar by 1 part in 3937. We have developed a method which makes improved use of the available MRP to remove interferences, and used it to obtain HCl-free 36Ar measurements on a multicollector instrument with MRP of only ~6000 (MRP= mass resolving power = m/dm 5-95% on side of peak). By arranging that the target mass position on a minor isotope (e.g. 36Ar), from which the interference must be removed, coincides with the ~50% point on the side of a major isotope (e.g. 40Ar), it is possible both to set the mass accurately and to verify the mass position and stability during measurements. The peak top of 40Ar is measured in a separate mass step. Two small corrections are necessary. One compensates for the residual HCl tail at the 36Ar position. The other arises because the peak is not totally flat in the region of interest: 40Ar and 36Ar+HCl are measured on the peak top, whilst 36Ar is measured at the extreme edge, with slightly lower efficiency. The required correction parameters can be obtained from a series of air calibrations with different target/interference ratios. With samples containing 4x10-15to 3x10-14moles of 40Ar, 36Ar/40Ar was measured, without HCl interference, to a 1σ precision of 0.5%, only slightly worse than counting statistics. This is potentially useful for 40Ar/39Ar dating, where 36Ar is used to correct for trapped air, and may be particularly significant for smaller or younger samples.

  3. Recominant Pinoresino-Lariciresinol Reductase, Recombinant Dirigent Protein And Methods Of Use

    DOEpatents

    Lewis, Norman G.; Davin, Laurence B.; Dinkova-Kostova, Albena T.; Fujita, Masayuki , Gang; David R. , Sarkanen; Simo , Ford; Joshua D.

    2003-10-21

    Dirigent proteins and pinoresinol/lariciresinol reductases have been isolated, together with cDNAs encoding dirigent proteins and pinoresinol/lariciresinol reductases. Accordingly, isolated DNA sequences are provided from source species Forsythia intermedia, Thuja plicata, Tsuga heterophylla, Eucommia ulmoides, Linum usitatissimum, and Schisandra chinensis, which code for the expression of dirigent proteins and pinoresinol/lariciresinol reductases. In other aspects, replicable recombinant cloning vehicles are provided which code for dirigent proteins or pinoresinol/lariciresinol reductases or for a base sequence sufficiently complementary to at least a portion of dirigent protein or pinoresinol/lariciresinol reductase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding dirigent protein or pinoresinol/lariciresinol reductase. Thus, systems and methods are provided for the recombinant expression of dirigent proteins and/or pinoresinol/lariciresinol reductases.

  4. Recombinant pinoresinol/lariciresinol reductase, recombinant dirigent protein, and methods of use

    DOEpatents

    Lewis, Norman G.; Davin, Laurence B.; Dinkova-Kostova, Albena T.; Fujita, Masayuki; Gang, David R.; Sarkanen, Simo; Ford, Joshua D.

    2001-04-03

    Dirigent proteins and pinoresinol/lariciresinol reductases have been isolated, together with cDNAs encoding dirigent proteins and pinoresinol/lariciresinol reductases. Accordingly, isolated DNA sequences are provided which code for the expression of dirigent proteins and pinoresinol/lariciresinol reductases. In other aspects, replicable recombinant cloning vehicles are provided which code for dirigent proteins or pinoresinol/lariciresinol reductases or for a base sequence sufficiently complementary to at least a portion of dirigent protein or pinoresinol/lariciresinol reductase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding dirigent protein or pinoresinol/lariciresinol reductase. Thus, systems and methods are provided for the recombinant expression of dirigent proteins and/or pinoresinol/lariciresinol reductases.

  5. 76 FR 27077 - Agency Information Collection Activities: Form AR-11 and Form AR-11SR, Extension of an Existing...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-10

    ... SECURITY U.S. Citizenship and Immigration Services Agency Information Collection Activities: Form AR-11 and...; OMB Control No. 1615-0007. The Department of Homeland Security, U.S. Citizenship and Immigration... collection: Form AR-11 and Form AR-11SR. U.S. Citizenship and Immigration Services. (4) Affected public...

  6. Paleotemperatures at the lunar surfaces from open system behavior of cosmogenic 38Ar and radiogenic 40Ar

    DOE PAGES

    Shuster, David L.; Cassata, William S.

    2015-02-10

    The simultaneous diffusion of both cosmogenic 38Ar and radiogenic 40Ar from solid phases is controlled by the thermal conditions of rocks while residing near planetary surfaces. Combined observations of 38Ar/37Ar and 40Ar/39Ar ratios during stepwise degassing analyses of neutron-irradiated Apollo samples can distinguish between diffusive loss of Ar due to solar heating of the rocks and that associated with elevated temperatures during or following impact events; the data provide quantitative constraints on the durations and temperatures of each process. From sequentially degassed 38Ar/37Ar ratios can be calculated a spectrum of apparent 38Ar exposure ages versus the cumulative release fraction ofmore » 37Ar, which is particularly sensitive to conditions at the lunar surface typically over ~106–108 year timescales. Due to variable proportions of K- and Ca-bearing glass, plagioclase and pyroxene, with variability in the grain sizes of these phases, each sample will have distinct sensitivity to, and therefore different resolving power on, past near-surface thermal conditions. Furthermore, we present the underlying assumptions, and the analytical and numerical methods used to quantify the Ar diffusion kinetics in multi-phase whole-rock analyses that provide these constraints.« less

  7. Kinetic measurements of phosphoglucose isomerase and phosphomannose isomerase by direct analysis of phosphorylated aldose-ketose isomers using tandem mass spectrometry

    NASA Astrophysics Data System (ADS)

    Gao, Hong; Chen, Ye; Leary, Julie A.

    2005-02-01

    A mass spectrometry based method for the direct determination of kinetic constants for phosphoglucose isomerase (PGI) and phosphomannose isomerase (PMI) is described. PGI catalyzes the interconversion between glucose-6-phosphate (Glc6P) and fructose-6-phosphate (Fru6P) and PMI performs the same function between mannose-6-phosphate (Man6P) and Fru6P. These two enzymes are essential in the pathways of glycolytic or oxidative metabolism of carbohydrates and have been considered as potential therapeutic targets. Traditionally, they are assayed either by spectrophotometric detection of Glc6P with one or more coupling enzymes or by a colorimetric detection of Fru6P. However, no suitable assay for Man6P has been developed yet to study the reaction of PMI in the direction from Fru6P to Man6P. In the work presented herein, a general assay for the isomeric substrate-product pair between Glc6P and Fru6P or between Man6P and Fru6P was developed, with the aim of directly studying the kinetics of PGI and PMI in both directions. The 6-phosphorylated aldose and ketose isomers were distinguished based on their corresponding tandem mass spectra (MS2) obtained on a quadrupole ion trap mass spectrometer, and a multicomponent quantification method was utilized to determine the composition of binary mixtures. Using this method, the conversion between Fru6P and Glc6P and that between Fru6P and Man6P are directly monitored. The equilibrium constants for the reversible reactions catalyzed by PGI and PMI are measured to be 0.3 and 1.1, respectively, and the kinetic parameters for both substrates of PGI and PMI are also determined. The values of KM and Vmax for Fru6P as substrate of PMI are reported to be 0.15 mM and 7.78 [mu]mol/(min mg), respectively. All other kinetic parameters measured correlate well with those obtained using traditional methods, demonstrating the accuracy and reliability of this assay.

  8. Dynamics and kinetics of the metastable decay series: Ar/sub 3//sup +/*. -->. Ar/sub 2//sup +/*. -->. Ar/sup +/

    SciTech Connect

    Scheier, P.; Stamatovic, A.; Maerk, T.D.

    1988-07-01

    Metastable decay of Ar/sup +//sup *//sub 3/ produced by electron impact ionization of a supersonic Ar cluster beam has been investigated qualitatively and quantitatively with a double focusing sector field mass spectrometer. We present direct experimental proof that the observed metastable decay of Ar/sup +//sup *//sub 3/ into Ar/sup +/ in the microsecond time regime proceeds predominantly via a sequential decay series Ar/sup +//sup *//sub 3/..-->../sup k//sup =/Ar/sup +//sup *//sub 2/..-->.. /sup k//sup =/Ar/sup +/ involving the evaporation of a single monomer in each of these successive decays. The metastable decay rates deduced are very different, i.e., k/sub 3,2/ = 45 s/sup -1/ and k/sub 2,1/ = 680 s/sup -1/ at an ion acceleration voltage of 3 kV. Moreover, the metastable decay rates of Ar/sup +//sup *//sub 3/ exhibit a strong dependence on time elapsed since electron impact production of the Ar/sup +//sub 3/, and on ion production parameters (i.e., size and internal excitation energy of the neutral precursor). For instance photodissociation (with visible laser light) of the neutral cluster beam leads to a dramatic increase of the observed metastable reaction rules yielding values of up to 100 000 3/sup -1/ in the ..mu..s time regime.

  9. Direct dating of weathering phenomena by [sup 40]Ar/[sup 39]Ar and K-Ar analysis of supergene K-Mn oxides

    SciTech Connect

    Vasconcelos, P.M.; Brimhall, G.H. ); Renne, P.R.; Becker, T.A. )

    1994-03-01

    Potassium-bearing manganese oxides, cryptomelane, K[sub 1-2](Mn[sup 3+]Mn[sup 4+])[sub 8] O[sub 16] [center dot] xH[sub 2]O, and hollandite, (K,Ba)[sub 1-2](Mn[sup 3+],Mn[sup 4+])[sub 8] O[sub 16] [center dot] xH[sub 2]O, are often authigenically precipitated in weathering profiles. Dating of these phases allows timing of the progression of oxidation fronts during weathering and pedogenic processes. Potential problems in manganese oxide dating, such as Ar and/or K losses, excess argon, [sup 39]Ar loss by recoil during neutron irradiation, etc. are addressed. The K-Ar and [sup 40]Ar/[sup 39]Ar analytical results indicate that Ar and/or K losses, excess [sup 40]Ar, and [sup 39]Ar recoil seem not to pose problems in manganese oxide dating. This investigation suggests that the fine scale, laser-probe [sup 40]Ar/[sup 39]Ar technique is most appropriate for dating of weathering phenomena because this technique permits identification of contaminating phases and the presence of multiple generations of weathering minerals in the inherently complex mineral assemblage characteristic of weathering profiles. K-Ar and [sup 40]Ar/[sup 39]Ar dating of supergene K-bearing manganese oxides formed during lateritization of Archean and Proterozoic bedrocks in the Carajas Region, Amazonia, Brazil, indicates that weathering started before 72 [+-] 6 Ma. Petrographic, electron microscope, and electron microprobe investigation reveal multiple generations of manganese oxide precipitation. Age clusters at 65-69, 51-56, 40-43, 33-35, 20, 24, 12-17 Ma, and zero-age (0.2 [+-] 0.2 Ma) suggest episodic precipitation of K-Mn oxides resulting form changing weathering conditions in the Amazon throughout the Cenozoic. K-Ar and [sup 40]Ar/[sup 39]Ar dating of supergene cryptomelane from weathering profiles in eastern Minas Gerais, southeastern Brazil, suggests continuous weathering from 10 to 5.6 Ma ago, possibly reflecting local climatic conditions due to the proximity with the Atlantic Ocean.

  10. Potential energy surface and bound states of the NH3-Ar and ND3-Ar complexes.

    PubMed

    Loreau, J; Liévin, J; Scribano, Y; van der Avoird, A

    2014-12-14

    A new, four-dimensional potential energy surface for the interaction of NH3 and ND3 with Ar is computed using the coupled-cluster method with single, double, and perturbative triple excitations and large basis sets. The umbrella motion of the ammonia molecule is explicitly taken into account. The bound states of both NH3-Ar and ND3-Ar are calculated on this potential for total angular momentum values from J = 0 to 10, with the inclusion of Coriolis interactions. The energies and splittings of the rovibrational levels are in excellent agreement with the extensive high-resolution spectroscopic data accumulated over the years in the infrared and microwave regions for both complexes, which demonstrates the quality of the potential energy surface.

  11. Ar-Ar and I-Xe Ages and the Thermal History of IAB Meteorites

    NASA Technical Reports Server (NTRS)

    Bogard, Donald D.; Garrison, Daniel H.; Takeda, Hiroshi

    2004-01-01

    Studies of several samples of the large Caddo County IAB iron meteorite reveal andesitic material, enriched in Si, Na, Al and Ca, which is essentially unique among meteorites. This material is believed to have formed from a chondritic source by partial melting and to have further segregated by grain coarsening. Such an origin implies extended metamorphism of the IAB parent body. New Ar-39-Ar-40 ages for silicate from three different Caddo samples are consistent with a common age of 4.50-4.51 Gyr ago. Less well defined Ar-Ar degassing ages for inclusions from two other IABs, EET8333 and Udei Station, are approx.4.32 Gyr, whereas the age for Campo del Cielo varies considerably over approx.3.23-4.56 Gyr. New I-129-Xe-129 ages for Caddo County and EET8333 are 4561.9+/-0.1 Myr and 4560- 4563 Myr, respectively, relative to an age of 4566 Myr for Shallowater. Considering all reported Ar-Ar ages for IABs and related winonaites, the range is approx.4.32-4.53 Gyr, but several IABs give similar Ar ages of 4.50-4.52 Gyr. We interpret these older ages to represent cooling after the time of last significant metamorphism on the parent body, and the younger ages to represent later Ar-40 diffusion loss. These older Ar-Ar ages are similar to Sm-Nd and Rb-Sr isochron ages reported in the literature for Caddo County. Considering the possibility that IAB parent body formation was followed by impact disruption, reassembly, and metamorphism (e.g., Benedix et al. 2000), the time of the post-assembly metamorphism may have been as late as approx.4.53 Gyr ago. However, precise I-Xe ages reported for some IABs define a range of ages of approx.4560 to approx.4576 Myr. The older I-Xe ages exceed the oldest precise radiometric ages of meteorites, appear unrealistic, and s,uggest a bias in the calibration of all I-Xe ages. But even with such a bias, the I-Xe ages of IABs cannot easily be reconciled with the much younger Ar-Ar and Sm-Nd ages and with cooling rates deduced from Ni concentration

  12. Ar-Ar and I-XE Ages and the Thermal History of IAB Meteorites

    NASA Technical Reports Server (NTRS)

    Bogard, Donald D.; Garrison, Daniel H.; Takeda, Hiroshi

    2006-01-01

    Studies of several samples of the large Caddo County IAB iron meteorite reveal andesitic material, enriched in Si, Na, Al and Ca which is essentially unique among meteorites. This material is believed to have formed from a chondritic source by partial melting and to have further segregated by grain coarsening. Such an origin implies extended metamorphism of the IAB parent body. New Ar-39- Ar-40 ages for silicate from three different Caddo samples are consistent with a common age of 4.50- 4.51 Gyr ago. Less well defined Ar-Ar degassing ages for inclusions from two other IABs, EET8333 and Udei Station, are approx. 4.32 Gyr, whereas the age for Campo del Cielo varies considerably over approx. 3.23-4.56 Gyr. New I-129-Xe-129 ges for Caddo County and EET8333 are 4561.9 plus or minus 0.1 Myr and 4560-4563 Myr, respectively, relative to an age of 4566 Myr for Shallowater. Considering all reported Ar-Ar ages for IABs and related winonaites, the range is approx. 4.32-4.53 Gyr, but several IABs give similar Ar ages of 4.50-4.52 Gyr. We interpret these older ages to represent cooling after the time of last significant metamorphism on the parent body, and the younger ages to represent later 40Ar diffusion loss. These older Ar-Ar ages are similar to Sm-Nd and Rb-Sr isochron ages reported in the literature for Caddo County. Considering the possibility that IAB parent body formation was followed by impact disruption, reassembly, and metamorphism (e.g., Benedix et al. 2000), the time of the postassembly metamorphism may have been as late as approx. 4.53 Gyr ago. However, precise I-Xe ages reported for some IABs define a range of ages of approx. 4560 to approx. 4576 Myr. The older I-Xe ages exceed the oldest precise radiometric ages of meteorites, appear unrealistic, and suggest a bias in the calibration of all I-Xe ages. But even with such a bias, the I-Xe ages of IABs cannot easily be reconciled with the much younger Ar-Ar and Sm-Nd ages and with cooling rates deduced from Ni

  13. Ar-Ar and I-Xe Ages and the Thermal History of IAB Meteorites

    NASA Technical Reports Server (NTRS)

    Bogard, Donald D.; Garrison, Daniel H.; Takeda, Hiroshi

    2004-01-01

    Studies of several samples of the large Caddo County IAB iron meteorite reveal andesitic material, enriched in Si, Nay Al and Ca, which is essentially unique among meteorites. This material is believed to have formed from a chondritic source by partial melting and to have further segregated by grain coarsening. Such an origin implies extended metamorphism of the IAB parent body. New Ar-39-Ar-40 ages for silicate from three different Caddo samples are consistent with a common age of 4.50- 4.51 Gyr ago. Less well defined Ar-Ar degassing ages for inclusions from two other IABs, EET8333 and Udei Station, are approx.4.32 Gyr, whereas the age for Campo del Cielo varies considerably over approx.3.23-4.56 Gyr. New I-129-Xe-129 ages for Caddo County and EET8333 are 4561.9 +/-0.1 Myr and 4560-4563 Myr, respectively, relative to an age of 4566 Myr for Shallowater. Considering all reported Ar-Ar ages for IABs and related winonaites, the range is approx.4.32-4.53 Gyr, but several IABs give similar Ar ages of 4.50-4.52 Gyr. We interpret these older ages to represent cooling after the time of last significant metamorphism on the parent body, and the younger ages to represent later Ar-40 diffusion loss. These older Ar-Ar ages are similar to Sm-Nd and Rb-Sr isochron ages reported in the literature for Caddo County. Considering the possibility that IAB parent body formation was followed by impact disruption, reassembly, and metamorphism (e.g., Benedix et al. 2000), the time of the post-assembly metamorphism may have been as late as approx.4.53 Gyr ago. However, precise I-Xe ages reported for some IABs define a range of ages of approx.4560 to approx.4576 My. The older I-Xe ages exceed the oldest precise radiometric ages of meteorites, appear unrealistic, and suggest a bias in the calibration of all I-Xe ages. But even with such a bias, the I-Xe ages of IABs cannot easily be reconciled with the much younger Ar-Ar and Sm-Nd ages and with cooling rates deduced from Ni concentration

  14. Investigation of the Gardnos Impact Structure: 40Ar/39Ar Results

    NASA Astrophysics Data System (ADS)

    Grier, J. A.; Swindle, T. D.; Kring, D. A.; Melosh, H. J.

    1995-09-01

    Introduction: The Gardnos structure (Norway) [1], has recently been identified as an impact crater [2,3], despite being deformed by a series of tectonic processes [2]. The age of the structure is uncertain, although attempts to date the impact event have been made on the basis of stratigraphic relationships [2,3]. Unfortunately, these estimates differ by more than 100 million years. The first study suggested that the impact corresponded roughly with the Cambro-Ordovician boundary, ~500 Ma [2], while a second stratigraphic interpretation suggested a Precambrian age of 650 Ma [3]. To try and resolve this discrepancy, we attempted to determine the age of the structure with radiometric techniques, which typically have an error of < 10 Ma. Method: We examined three samples from the crater, in an attempt to determine the age using the 40Ar/39Ar dating method. The samples (plagioclase feldspar, alkali feldspar, and shards of gray, semi-transparent, glassy-looking material) were obtained from the suevite breccia unit of [2]. This unit should be the most promising for 40Ar/39Ar dating, since it is melt bearing, and no other unit is likely to be less affected by subsequent tectonism and sedimentation. Results: Our experiments yielded complicated age spectra interpreted to reflect a thermal event at 385+/-5 Ma and another possible event at 312+/-5 Ma (Fig. 1). All three samples yielded similar results. If the stratigraphic interpretations are correct, it thus seems clear that the impact event itself was not dated. The 385 Ma age corresponds to the end of the Caledonian Orogeny, which significantly altered this section of Norway [4]. It is suspected that late Caledonian folding occurred during this time [4], and we therefore interpret the 385 Ma age as the thermal metamorphic overprint of the Caledonian Orogeny. Another event clearly occurred afterwards, which is responsible for the 312 Ma signature. The cause of this thermal overprint has not yet been identified, since

  15. Ar-Ar and Rb-Sr Ages of the Tissint Olivine-phyric Martian Shergottite

    NASA Technical Reports Server (NTRS)

    Park, J.; Herzog, G. F.; Nyquist, L. E.; Shih, C.-Y.; Turin, B.; Lindsay, F. N.; Delaney, J. S.; Swisher, C. C., III; Agee, C.

    2013-01-01

    The fifth martian meteorite fall, Tissint, is an olivine-phyric shergottite that contains olivine macrocrysts (approximately 1.5 mm) [1]. [2] reported the Sm-Nd age of Tissint as 596 plus or minus 23 Ma along with Rb-Sr data that defined no isochron. [3] reported Lu-Hf and Sm-Nd ages of 583 plus or minus 86 Ma and 616 plus or minus 67 Ma, respectively. The cosmic-ray exposure ages of Tissint are 1.10 plus or minus 0.15 Ma based on 10Be [4], and 1.0-1.1 Ma, based on 3He, 21Ne, and 38Ar [5,6].We report Ar-Ar ages and Rb-Sr data.

  16. Laser /39/Ar-/40/Ar dating of two clasts from consortium breccia 73215

    NASA Technical Reports Server (NTRS)

    Eichhorn, G.; Schaeffer, O. A.; James, O. B.; Mueller, H. W.

    1978-01-01

    A laser Ar-39-Ar-40 study of the components of an ANT-suite anorthositic gabbro and a black aphanite from a consortium breccia is reported. A wide range of K-Ar ages is found for the plagioclase in the anorthositic gabbro; at the centers of the largest grains is material showing the greatest age (older than 4.11 billion years) while the youngest material (3.81-3.88 billion years) is found near the grain margins. Partial outgassing of the clasts upon incorporation into the breccia could account for the age patterns. The black aphanite clast appears to be cogenetic with the aphanite that forms the breccia matrix. The time of crystallization of a lunar granite has also been measured by the laser technique.

  17. The X-ray crystal structure of APR-B, an atypical adenosine 5'-phosphosulfate reductase from Physcomitrella patens.

    PubMed

    Stevenson, Clare E M; Hughes, Richard K; McManus, Michael T; Lawson, David M; Kopriva, Stanislav

    2013-11-15

    Sulfonucleotide reductases catalyse the first reductive step of sulfate assimilation. Their substrate specificities generally correlate with the requirement for a [Fe4S4] cluster, where adenosine 5'-phosphosulfate (APS) reductases possess a cluster and 3'-phosphoadenosine 5'-phosphosulfate reductases do not. The exception is the APR-B isoform of APS reductase from the moss Physcomitrella patens, which lacks a cluster. The crystal structure of APR-B, the first for a plant sulfonucleotide reductase, is consistent with a preference for APS. Structural conservation with bacterial APS reductase rules out a structural role for the cluster, but supports the contention that it enhances the activity of conventional APS reductases.

  18. MEIS1 functions as a potential AR negative regulator

    SciTech Connect

    Cui, Liang; Yang, Yutao; Hang, Xingyi; Cui, Jiajun; Gao, Jiangping

    2014-10-15

    The androgen receptor (AR) plays critical roles in human prostate carcinoma progression and transformation. However, the activation of AR is regulated by co-regulators. MEIS1 protein, the homeodomain transcription factor, exhibited a decreased level in poor-prognosis prostate tumors. In this study, we investigated a potential interaction between MEIS1 and AR. We found that overexpression of MEIS1 inhibited the AR transcriptional activity and reduced the expression of AR target gene. A potential protein–protein interaction between AR and MEIS1 was identified by the immunoprecipitation and GST pull-down assays. Furthermore, MEIS1 modulated AR cytoplasm/nucleus translocation and the recruitment to androgen response element in prostate specific antigen (PSA) gene promoter sequences. In addition, MEIS1 promoted the recruitment of NCoR and SMRT in the presence of R1881. Finally, MEIS1 inhibited the proliferation and anchor-independent growth of LNCaP cells. Taken together, our data suggests that MEIS1 functions as a novel AR co-repressor. - Highlights: • A potential interaction was identified between MEIS1 and AR signaling. • Overexpression of MEIS1 reduced the expression of AR target gene. • MEIS1 modulated AR cytoplasm/nucleus translocation. • MEIS1 inhibited the proliferation and anchor-independent growth of LNCaP cells.

  19. The 40Ar/39Ar geochronology of the Pelona schist and related rocks, southern California

    NASA Astrophysics Data System (ADS)

    Jacobson, Carl E.

    1990-01-01

    Seventeen 40Ar/39Ar ages for hornblende, celadonitic muscovite, and biotite from the Pelona, Orocopia, Rand, and Portal Ridge (POR) schists range from 39 to 85 Ma. The 85 Ma is from Portal Ridge, consistent with the proposed correlation of that body with the schist of Sierra de Salinas. Two muscovites and one hornblende from the Rand Schist have ages of 72 to 74 Ma, indistinguishable from the K-Ar age of 74 Ma for hornblende from a posttectonic granodiorite that intrudes the schist, but younger than the 79 Ma U-Pb age of the intrusion. Four muscovite and two hornblende ages for schist and mylonite from the East Fork area of the San Gabriel Mountains range from 55 to 61 Ma, similar to K-Ar ages from nearby upper plate rocks. A third hornblende has an age of 73 Ma but is not well constrained. Concordance of schist and upper plate ages confirms structural and metamorphic evidence that the Vincent thrust in the San Gabriel Mountains has not undergone significant postmetamorphic disruption. Ages from the Orocopia Mountains are 75 Ma for hornblende from nonmylonitic upper plate, 52 Ma for muscovite from structurally high Orocopia Schist that is mylonitic, and 41 Ma for muscovite from nonmylonitic Orocopia Schist. These are consistent with field evidence that the Orocopia "thrust" is a postmetamorphic normal fault. Muscovite and hornblende from the Gavilan Hills have ages of 48 to 50 Ma, younger than ages from the San Gabriel Mountains but similar to schist ages from the Orocopia Mountains. The geochronologic and structural complexities of the Vincent, Chocolate Mountains, Orocopia, and Rand thrusts imply that previously cited northeastward vergence may not relate to prograde metamorphism (subduction) of the POR schists. The 40Ar/39Ar data indicate substantial uplift of the POR schists prior to middle Tertiary detachment faulting, which confirms other geochronologic evidence of uplift in southern California and southern Arizona during the Late Cretaceous-early Tertiary.

  20. [sup 40]Ar/[sup 39]Ar thermochronology in the northern Bitterroot mylonite zone, Mt

    SciTech Connect

    House, M.A.; Hodges, K.V. . Dept. of Earth, Atmospheric, and Planetary Sciences)

    1993-04-01

    The extensional Bitterroot mylonite zone defines the eastern and southern border of the Bitterroot metamorphic core complex and is generally interpreted to be the major structure which accommodated unroofing of the metamorphic core. The most commonly cited evidence for the age of mylonitization are [sup 40]Ar/[sup 39]Ar ages for hornblend, muscovite, biotite, and potassium feldspar from the southern Bitterroot mylonite zone that indicate rapid cooling of the core rocks between 45.5 and 43.5 Ma. More recently, an [sup 40]Ar/[sup 39]Ar K-feldspar age of 46.4 [+-] 0.8 Ma for an undeformed rhyolite dike that cuts across the mylonitic fabric places a minimum age constraint on the southern part of the shear zone. The authors have obtained new [sup 40]Ar/[sup 39]Ar data for metapelitic rocks and amphibolites from the northeast border of the Bitterroot metamorphic core complex near an area where mylonitized granitoid rocks yielding 48--52 Ma U-Pb zircon crystallization ages constrain the maximum age of mylonitization. Isochran ages of 47.9 [+-] 0.9 and 49 [+-] 1 Ma for hornblende separated from deformed amphibolite pods in the northeast border zone are within analytical uncertainty of the younger mylonitized granitoid crystallization ages and indicate rapid post-crystallization cooling through temperatures of [approximately]780--800 K. They attribute this cooling to denudation related to shear zone development. Muscovite and biotite isochron ages from metapelitic rocks within the shear zone are significantly younger, between 42 and 44 Ms., and generally agree with mica ages obtained by Garmezy and Sutter for the southern part of the shear zone. However, all mica ages from the Bitterroot shear zone are younger than the minimum age of the shear zone deduced from the age of cross-cutting rhyolite dikes.

  1. 40Ar/39Ar and cosmic ray exposure ages of plagioclase-rich lithic fragments from Apollo 17 regolith, 78461

    NASA Astrophysics Data System (ADS)

    Das, J. P.; Baldwin, S. L.; Delano, J. W.

    2016-01-01

    Argon isotopic data is used to assess the potential of low-mass samples collected by sample return missions on planetary objects (e.g., Moon, Mars, asteroids), to reveal planetary surface processes. We report the first 40Ar/39Ar ages and 38Ar cosmic ray exposure (CRE) ages, determined for eleven submillimeter-sized (ranging from 0.06 to 1.2 mg) plagioclase-rich lithic fragments from Apollo 17 regolith sample 78461 collected at the base of the Sculptured Hills. Total fusion analysis was used to outgas argon from the lithic fragments. Three different approaches were used to determine 40Ar/39Ar ages and illustrate the sensitivity of age determination to the choice of trapped (40Ar/36Ar)t. 40Ar/39Ar ages range from ~4.0 to 4.4 Ga with one exception (Plag#10). Surface CRE ages, based on 38Ar, range from ~1 to 24 Ma. The relatively young CRE ages suggest recent re-working of the upper few centimeters of the regolith. The CRE ages may result from the effect of downslope movement of materials to the base of the Sculptured Hills from higher elevations. The apparent 40Ar/39Ar age for Plag#10 is >5 Ga and yielded the oldest CRE age (i.e., ~24 Ma). We interpret this data to indicate the presence of parentless 40Ar in Plag#10, originating in the lunar atmosphere and implanted in lunar regolith by solar wind. Based on a chemical mixing model, plagioclase compositions, and 40Ar/39Ar ages, we conclude that lithic fragments originated from Mg-suite of highland rocks, and none were derived from the mare region.

  2. Ar-Ar Dating of Martian Chassignites, NWA2737 and Chassigny, and Nakhlite MIL03346

    NASA Technical Reports Server (NTRS)

    Bogard, D. D.; Garrison, D. H.

    2006-01-01

    Until recently only three nakhlites and one chassignite had been identified among martian meteorites. These four exhibit very similar radiometric ages and cosmic ray exposure (CRE) ages, indicating that they may have derived from a common location on Mars and were ejected into space by a single impact. This situation is quite different from that of martian shergottites, which exhibit a range of radiometric ages and CRE ages (1). Recently, several new nakhlites and a new martian dunite (NWA2737) have been recognized. Here we report our results of Ar-39-Ar-40 dating for the MIL03346 nakhlite and the NWA2737 "chassignite", along with new results on Chassigny.

  3. The barents sea magmatic province: Geological-geophysical evidence and new 40Ar/39Ar dates

    NASA Astrophysics Data System (ADS)

    Shipilov, E. V.; Karyakin, Yu. V.

    2011-07-01

    Resulting from study of the geological structure of the Franz Josef Land and Svalbard archipelagoes, this work presents new 17 40Ar/39Ar age datings for basalts taken during coastal expeditions in 2006-2010. Radiological age determination for intrusive units (sills) located in the western part of Nordensciold Land (Spitzbergen Island) has been made for the first time. In relation to use of the interpretation results of marine geological-geophysical data, the distribution peculiarities and time ranges for Jurassic-Cretaceous basic magmatism within the studied regions of the Barents Sea continental margin and within the Arctic as a whole are discussed.

  4. SUMER-IRIS Observations of AR11875

    NASA Astrophysics Data System (ADS)

    Schmit, Donald; Innes, Davina

    2014-05-01

    We present results of the first joint observing campaign of IRIS and SOHO/SUMER. While the IRIS datasets provide information on the chromosphere and transition region, SUMER provides complementary diagnostics on the corona. On 2013-10-24, we observed an active region, AR11875, and the surrounding plage for approximately 4 hours using rapid-cadence observing programs. These datasets include spectra from a small C -class flare which occurs in conjunction with an Ellerman-bomb type event. Our analysis focusses on how the high spatial resolution and slit jaw imaging capabilities of IRIS shed light on the unresolved structure of transient events in the SUMER catalog.

  5. The Spectroscopic study of {sup 33}Ar

    SciTech Connect

    Adimi, N.; Dominguez-Reyes, R.; Alcorta, M.; Borge, M. J. G.; Perea, A.; Tengblad, O.; Bey, A.; Blank, B.; Dossat, C.; Giovinazzo, J.; Matea, I.; Fynbo, H. O. U.; Knudsen, H. H.; Suemmerer, K.

    2011-10-28

    The proton-rich nucleus {sup 33}Ar has been produced at the low-energy facility SPIRAL at GANIL. Spectroscopic studies of gamma and p emission of this nucleus were performed with the 'Silicon Cube' detection system. The analysis of proton and gamma singles and coincidence spectra allowed us to establish a complete decay scheme of this nucleus. The comparison of the Gamow-Teller strength distribution deduced from our experiment and the theoretical one obtained with the Shell Model permitted the determination of a quenching factor for the Gamow-Teller strength.

  6. Theoretical photoabsorption spectra of Ar n+ clusters

    NASA Astrophysics Data System (ADS)

    Doltsinis, Nikos L.; Knowles, Peter J.

    2000-08-01

    The photoabsorption spectra of selected Ar n+ clusters ( n=7, 8, 17, 19, 23) have been investigated theoretically using an extended Diatomics-in-Molecules approach including induced dipole - induced dipole and spin-orbit coupling interaction effects. Our calculations at 0 K confirm the experimentally observed spectral red-shift of the visible photoabsorption peak in the region 15< n<20 [Levinger et al., J. Chem. Phys. 89 (1988) 5654]. Furthermore, we have been able to reproduce the additional red-shift measured for 7⩽ n⩽9 [Haberland et al., Phys. Rev. Lett. 67 (1991) 3290] by carrying out finite temperature Monte Carlo simulations.

  7. Codon 89 polymorphism in the human 5 α -reductase gene in primary breast cancer

    PubMed Central

    Scorilas, A; Bharaj, B; Giai, M; Diamandis, E P

    2001-01-01

    The enzyme human steroid 5-α reductase type II (SRD5A2) and androgen receptor (AR) are critical mediators of androgen action, suggesting a potential role in hormonally related cancers. The SRD5A2 gene harbours two frequent polymorphic sites, one in the coding region, at codon 89 of exon 1, where valine is substituted by leucine (V89L) and the other in the 3′ untranslated region (3′ UTR) where a variable number of dinucleotide TA repeat lengths exists. The V89L polymorphism is known to alter the activity of this enzyme. In the present study we examined 144 sporadic breast tumours from Italian patients for the V89L and TA polymorphisms by sequence and fragment analysis, respectively. Tumour extract prostate specific antigen (PSA) concentration as well as a number of well-established clinical and pathological parameters were evaluated. The results show that 53% of the tumours were homozygous for VV alleles, 37% were heterozygous for VL alleles and 10% were homozygous for LL alleles. TA(0) repeats were found in tumours with VV, LL and VL genotypes. TA(9) repeats were only found in VV homozygotes and were totally absent from either LL homozygotes or VL heterozygotes. PSA expression was significantly elevated in tumours with VV genotype. The presence of LL alleles in breast tumours is associated with earlier onset and shorter disease-free (RR = 2.65;P = 0.013) and overall survival (RR = 3.06;P = 0.014) rates. The VV genotype is associated with a more favourable prognosis. Our study suggests that the polymorphism in codon 89 of exon 1 of the human 5α-reductase gene is related with TA repeat genotypes, PSA expression and breast cancer prognosis. More specifically, we found that the LL genotype is also associated with earlier onset and more aggressive forms of breast cancer. Long-term-outcome studies are needed to investigate the relevance of this polymorphism to breast cancer susceptibility. © 2001 Cancer Research Campaign http://www.bjcancer.com PMID:11259089

  8. Genetic mapping of the interface between the ArsD metallochaperone and the ArsA ATPase

    PubMed Central

    Yang, Jianbo; Ajees, Abdul; Salam, Abdul; Rosen, Barry P.

    2011-01-01

    Summary The ArsD metallochaperone delivers trivalent metalloids, As(III) or Sb(III), to the ArsA ATPase, the catalytic subunit of the ArsAB As(III) efflux pump. Transfer of As(III) increases the affinity of ArsA for As(III), allowing resistance to environmental arsenic concentrations. As(III) transfer is channeled from chaperone to ATPase, implying that ArsD and ArsA form an interface at their metal binding sites. A genetic approach was used to test this hypothesis. Thirteen ArsD mutants exhibiting either weaker or stronger interaction with ArsA were selected by either repressed transactivator yeast two-hybrid or reverse yeast two-hybrid assays. Additionally, Lys-37 and Lys-62 were identified as being involved in ArsD function by site-directed mutagenesis and chemical modification. Substitution at either position with arginine was tolerated, suggesting participation of a positive charge. By yeast two-hybrid analysis K37A and K62A mutants lost interaction with ArsA. All fifteen mutations were mapped on the surface of the ArsD structure, and their locations are consistent with a structural model generated by in silico docking. Four are close to metalloid binding site residues Cys-12, Cys-13 and Cys18, and seven are on the surface of helix 1. These results suggest that the interface involves one surface of helix 1 and the metalloid binding site. PMID:21299644

  9. Gonadotropin-regulated testicular RNA helicase (GRTH/DDX25), a negative regulator of luteinizing/chorionic gonadotropin hormone-induced steroidogenesis in Leydig cells: central role of steroidogenic acute regulatory protein (StAR).

    PubMed

    Fukushima, Masato; Villar, Joaquin; Tsai-Morris, Chon-Hwa; Dufau, Maria L

    2011-08-26

    Gonadotropin-regulated testicular RNA helicase (GRTH/DDX25) is a testis-specific gonadotropin-regulated RNA helicase that is present in Leydig cells (LCs) and germ cells and is essential for spermatid development and completion of spermatogenesis. Normal basal levels of testosterone in serum and LCs were observed in GRTH null (GRTH(-/-)) mice. However, testosterone production was enhanced in LCs of GRTH(-/-) mice compared with WT mice by both in vivo and in vitro human chorionic gonadotropin stimulation. LCs of GRTH(-/-) mice had swollen mitochondria with a significantly increased cholesterol content in the inner mitochondrial membrane. Basal protein levels of SREBP2, HMG-CoA reductase, and steroidogenic acute regulatory protein (StAR; a protein that transports cholesterol to the inner mitochondrial membrane) were markedly increased in LCs of GRTH(-/-) mice compared with WT mice. Gonadotropin stimulation caused an increase in StAR mRNA levels and protein expression in GRTH(-/-) mice versus WT mice, with no further increase in SREBP2 and down-regulation of HMG-CoA reductase protein. The half-life of StAR mRNA was significantly increased in GRTH(-/-) mice. Moreover, association of StAR mRNA with GRTH protein was observed in WT mice. Human chorionic gonadotropin increased GRTH gene expression and its associated StAR protein at cytoplasmic sites. Taken together, these findings indicate that, through its negative role in StAR message stability, GRTH regulates cholesterol availability at the mitochondrial level. The finding of an inhibitory action of GRTH associated with gonadotropin-mediated steroidogenesis has provided insights into a novel negative autocrine molecular control mechanism of this helicase in the regulation of steroid production in the male.

  10. The role of mitochondrial fusion and StAR phosphorylation in the regulation of StAR activity and steroidogenesis.

    PubMed

    Castillo, Ana F; Orlando, Ulises; Helfenberger, Katia E; Poderoso, Cecilia; Podesta, Ernesto J

    2015-06-15

    The steroidogenic acute regulatory (StAR) protein regulates the rate-limiting step in steroidogenesis, i.e. the delivery of cholesterol from the outer (OMM) to the inner (IMM) mitochondrial membrane. StAR is a 37-kDa protein with an N-terminal mitochondrial targeting sequence that is cleaved off during mitochondrial import to yield 30-kDa intramitochondrial StAR. StAR acts exclusively on the OMM and its activity is proportional to how long it remains on the OMM. However, the precise fashion and the molecular mechanism in which StAR remains on the OMM have not been elucidated yet. In this work we will discuss the role of mitochondrial fusion and StAR phosphorylation by the extracellular signal-regulated kinases 1/2 (ERK1/2) as part of the mechanism that regulates StAR retention on the OMM and activity.

  11. The respiratory arsenate reductase from Bacillus selenitireducens strain MLS10

    USGS Publications Warehouse

    Afkar, E.; Lisak, J.; Saltikov, C.; Basu, P.; Oremland, R.S.; Stolz, J.F.

    2003-01-01

    The respiratory arsenate reductase from the Gram-positive, haloalkaliphile, Bacillus selenitireducens strain MLS10 was purified and characterized. It is a membrane bound heterodimer (150 kDa) composed of two subunits ArrA (110 kDa) and ArrB (34 kDa), with an apparent Km for arsenate of 34 ??M and Vmax of 2.5 ??mol min-1 mg-1. Optimal activity occurred at pH 9.5 and 150 g l-1 of NaCl. Metal analysis (inductively coupled plasma mass spectrometry) of the holoenzyme and sequence analysis of the catalytic subunit (ArrA; the gene for which was cloned and sequenced) indicate it is a member of the DMSO reductase family of molybdoproteins. ?? 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

  12. Involvement of nitrate reductase in auxin-induced NO synthesis

    PubMed Central

    Erdei, L

    2008-01-01

    It is well known for a long time, that nitric oxide (NO) functions in variable physiological and developmental processes in plants, however the source of this signaling molecule in the diverse plant responses is very obscure.1 Although existance of nitric oxide sythase (NOS) in plants is still questionable, LNMMA (NG-monomethyl-L-arginine)-sensitive NO generation was observed in different plant species.2,3 In addition, nitrate reductase (NR) is confirmed to have a major role as source of NO.4,5 This multifaced molecule acts also in auxin-induced lateral root (LR) formation, since exogenous auxin enhanced NO levels in regions of Arabidopsis LR initiatives. Our results pointed out the involvement of nitrate reductase enzyme in auxin-induced NO formation. In this addendum, we speculate on auxin-induced NO production in lateral root primordial formation. PMID:19704423

  13. Involvement of nitrate reductase in auxin-induced NO synthesis.

    PubMed

    Kolbert, Zsuzsanna; Erdei, L

    2008-11-01

    It is well known for a long time, that nitric oxide (NO) functions in variable physiological and developmental processes in plants, however the source of this signaling molecule in the diverse plant responses is very obscure.1 Although existance of nitric oxide sythase (NOS) in plants is still questionable, LNMMA (N(G)-monomethyl-L-arginine)-sensitive NO generation was observed in different plant species.2,3 In addition, nitrate reductase (NR) is confirmed to have a major role as source of NO.4,5 This multifaced molecule acts also in auxin-induced lateral root (LR) formation, since exogenous auxin enhanced NO levels in regions of Arabidopsis LR initiatives. Our results pointed out the involvement of nitrate reductase enzyme in auxin-induced NO formation. In this addendum, we speculate on auxin-induced NO production in lateral root primordial formation. PMID:19704423

  14. New high-precision 40Ar/39Ar ages on Oligocene volcanic rocks of northwestern Kenya

    NASA Astrophysics Data System (ADS)

    Brown, Francis H.; Jicha, Brian R.

    2016-02-01

    New, high-precision 40Ar/39Ar ages from volcanic rocks in northwestern Kenya are provided for some areas of exposure in this remote area. We report seven 40Ar/39Ar ages generated from single crystal total fusion experiments on alkali feldspar separated from volcanic rocks in the Mogila, Songot, and Lokwanamur Ranges and the Gatome valley. A rhyolite from the lower part of the sequence in the Mogila Range yielded ages of 32.31 ± 0.06 Ma and 32.33 ± 0.07 Ma, and a rhyolite near the top of that sequence yielded 31.67 ± 0.04 Ma. A single sample from the Songot Range yielded an age of 32.49 ± 0.07 Ma, slightly older than the rocks collected from Mogila. In both ranges the early Oligocene rhyolites are underlain by basalts, as is also the case in the Labur Range. Ages of 25.95 ± 0.03 Ma, 25.91 ± 0.04 Ma, and 27.15 ± 0.03 Ma were measured on alkali feldspar from rhyolites from the Lokwanamur Range, and the nearby Gatome valley. All of these rocks are part of an episode of widespread volcanism in northwestern Kenya in the mid-to late Oligocene that is not currently known from the Ethiopian Rift Valley.

  15. 40Ar/39Ar age of material returned from asteroid 25143 Itokawa

    NASA Astrophysics Data System (ADS)

    Park, Jisun; Turrin, Brent D.; Herzog, Gregory F.; Lindsay, Fara N.; Delaney, Jeremy S.; Swisher, Carl C.; Uesugi, Masayuki; Karouji, Yuzuru; Yada, Toru; Abe, Masanao; Okada, Tatsuaki; Ishibashi, Yukihiro

    2015-11-01

    The Hayabusa mission to asteroid 25143, Itokawa, brought back 2000 small particles, which most closely resemble material found in LL4-6 chondrites. We report an 40Ar/39Ar age of 1.3 ± 0.3 Ga for a sample of Itokawa consisting of three grains with a total mass of ~2 μg. This age is lower than the >4.0 Ga ages measured for 75% of LL chondrites but close to one for Y-790964 and its pairs. The flat 40Ar/39Ar release spectrum of the sample suggests complete degassing 1.3 Ga ago. Recent solar heating in Itokawa's current orbit does not appear likely to have reset that age. Solar or impact heating 1.3 Ga ago could have done so. If impact heating was responsible, then the 1.3 Ga age sets an upper bound on the time at which the Itokawa rubble pile was assembled and suggests that rubble pile creation was an ongoing process in the inner solar system for at least the first 3 billion years of solar system history.

  16. Ar40-Ar39 systematics in rocks and separated minerals from Apollo 14.

    NASA Technical Reports Server (NTRS)

    Turner, G.; Huneke, J. C.; Podosek, F. A.; Wasserburg, G. J.

    1972-01-01

    The Ar40-Ar39 dating technique has been applied to separated minerals (plagioclase, pyroxene, quintessence and an 'ilmenite' concentrate), and whole rock samples of Apollo 14 rocks 14310 and 14073. Plagioclase shows the best gas retention characteristics, with no evidence of anomalous behavior and only a small amount of gas loss in the initial release. Ages determined from the plagioclase of 14310 and 14073 are (3.87 plus or minus 0.05) and (3.88 plus or minus 0.05) AE respectively. Low apparent ages at low release temperatures, which are frequently observed in whole rock Ar40-Ar39 experiments on lunar basalts, are shown to be principally due to gas loss in the high-K interstitial glass (quintessence) phase, confirming earlier suggestions. The decrease in apparent ages in the high-temperature release previously observed in several total rock samples of Apollo 14 basalts has been identified with the pyroxene. Plagioclase is also found to be the most suitable mineral for the determination of cosmic ray exposure ages, and exposure ages of 280 and 113 m.y. are found for 14310 and 14073, respectively, indicating that these rocks, which are very similar in many respects, have different exposure histories.

  17. Using chemical approaches to study selenoproteins - focus on thioredoxin reductases

    PubMed Central

    Hondal, Robert J.

    2009-01-01

    The study of selenocysteine-containing proteins is difficult due to the problems associated with the heterologous production of these proteins. These problems are due to the intricate recoding mechanism used by cells to translate the UGA codon as a sense codon for selenocysteine. The process is further complicated by the fact that eukaryotes and prokaryotes have different UGA recoding machineries. This review focuses on chemical approaches to produce selenoproteins and study the mechanism of selenoenzymes. The use of intein-mediated peptide ligation is discussed with respect to the production of the mammalian selenoenzymes thioredoxin reductase and selenoprotein R, also known as methionine sulfoxide reductase B1. New methods for removing protecting groups from selenocysteine post-synthesis and methods for selenosulfide/diselenide formation are also reviewed. Chemical approaches have also been used to study the enzymatic mechanism of thioredoxin reductase. The approach divides the enzyme into two modules, a large protein module lacking selenocysteine and a small, synthetic selenocysteine-containing peptide. Study of this semisynthetic enzyme has revealed three distinct enzymatic pathways that depend on the properties of the substrate. The enzyme utilizes a macromolecular mechanism for protein substrates, a second mechanism for small molecule substrates and a third pathway for selenium-containing substrates such as selenocystine. PMID:19406205

  18. Cloning and sequence of the human adrenodoxin reductase gene.

    PubMed Central

    Lin, D; Shi, Y F; Miller, W L

    1990-01-01

    Adrenodoxin reductase (ferrodoxin:NADP+ oxidoreductase, EC 1.18.1.2) is a flavoprotein mediating electron transport to all mitochondrial forms of cytochrome P450. We cloned the human adrenodoxin reductase gene and characterized it by restriction endonuclease mapping and DNA sequencing. The entire gene is approximately 12 kilobases long and consists of 12 exons. The first exon encodes the first 26 of the 32 amino acids of the signal peptide, and the second exon encodes the remainder of signal peptide and the apparent FAD binding site. The remaining 10 exons are clustered in a region of only 4.3 kilobases, separated from the first two exons by a large intron of about 5.6 kilobases. Two forms of human adrenodoxin reductase mRNA, differing by the presence or absence of 18 bases in the middle of the sequence, arise from alternate splicing at the 5' end of exon 7. This alternately spliced region is directly adjacent to the NADPH binding site, which is entirely contained in exon 6. The immediate 5' flanking region lacks TATA and CAAT boxes; however, this region is rich in G + C and contains six copies of the sequence GGGCGGG, resembling promoter sequences of "housekeeping" genes. RNase protection experiments show that transcription is initiated from multiple sites in the 5' flanking region, located about 21-91 base pairs upstream from the AUG translational initiation codon. Images PMID:2236061

  19. Structural and functional diversity of ferredoxin-NADP(+) reductases.

    PubMed

    Aliverti, Alessandro; Pandini, Vittorio; Pennati, Andrea; de Rosa, Matteo; Zanetti, Giuliana

    2008-06-15

    Although all ferredoxin-NADP(+) reductases (FNRs) catalyze the same reaction, i.e. the transfer of reducing equivalents between NADP(H) and ferredoxin, they belong to two unrelated families of proteins: the plant-type and the glutathione reductase-type of FNRs. Aim of this review is to provide a general classification scheme for these enzymes, to be used as a framework for the comparison of their properties. Furthermore, we report on some recent findings, which significantly increased the understanding of the structure-function relationships of FNRs, i.e. the ability of adrenodoxin reductase and its homologs to catalyze the oxidation of NADP(+) to its 4-oxo derivative, and the properties of plant-type FNRs from non-photosynthetic organisms. Plant-type FNRs from bacteria and Apicomplexan parasites provide examples of novel ways of FAD- and NADP(H)-binding. The recent characterization of an FNR from Plasmodium falciparum brings these enzymes into the field of drug design.

  20. Modulating hemoglobin nitrite reductase activity through allostery: a mathematical model.

    PubMed

    Rong, Zimei; Alayash, Abdu I; Wilson, Michael T; Cooper, Chris E

    2013-11-30

    The production of nitric oxide by hemoglobin (Hb) has been proposed to play a major role in the control of blood flow. Because of the allosteric nature of hemoglobin, the nitrite reductase activity is a complex function of oxygen partial pressure PO2. We have previous developed a model to obtain the micro rate constants for nitrite reduction by R state (kR) and T state (kT) hemoglobin in terms of the experimental maximal macro rate constant kNmax and the corresponding oxygen concentration PO2max. However, because of the intrinsic difficulty in obtaining accurate macro rate constant kN, from available experiments, we have developed an alternative method to determine the micro reaction rate constants (kR and kT) by fitting the simulated macro reaction rate curve (kN versus PO2) to the experimental data. We then use our model to analyze the effect of pH (Bohr Effect) and blood ageing on the nitrite reductase activity, showing that the fall of bisphosphoglycerate (BPG) during red cell storage leads to increase NO production. Our model can have useful predictive and explanatory power. For example, the previously described enhanced nitrite reductase activity of ovine fetal Hb, in comparison to the adult protein, may be understood in terms of a weaker interaction with BPG and an increase in the value of kT from 0.0087M(-1)s(-1) to 0.083M(-1)s(-1).

  1. Perchlorate Reductase Is Distinguished by Active Site Aromatic Gate Residues.

    PubMed

    Youngblut, Matthew D; Tsai, Chi-Lin; Clark, Iain C; Carlson, Hans K; Maglaqui, Adrian P; Gau-Pan, Phonchien S; Redford, Steven A; Wong, Alan; Tainer, John A; Coates, John D

    2016-04-22

    Perchlorate is an important ion on both Earth and Mars. Perchlorate reductase (PcrAB), a specialized member of the dimethylsulfoxide reductase superfamily, catalyzes the first step of microbial perchlorate respiration, but little is known about the biochemistry, specificity, structure, and mechanism of PcrAB. Here we characterize the biophysics and phylogeny of this enzyme and report the 1.86-Å resolution PcrAB complex crystal structure. Biochemical analysis revealed a relatively high perchlorate affinity (Km = 6 μm) and a characteristic substrate inhibition compared with the highly similar respiratory nitrate reductase NarGHI, which has a relatively much lower affinity for perchlorate (Km = 1.1 mm) and no substrate inhibition. Structural analysis of oxidized and reduced PcrAB with and without the substrate analog SeO3 (2-) bound to the active site identified key residues in the positively charged and funnel-shaped substrate access tunnel that gated substrate entrance and product release while trapping transiently produced chlorate. The structures suggest gating was associated with shifts of a Phe residue between open and closed conformations plus an Asp residue carboxylate shift between monodentate and bidentate coordination to the active site molybdenum atom. Taken together, structural and mutational analyses of gate residues suggest key roles of these gate residues for substrate entrance and product release. Our combined results provide the first detailed structural insight into the mechanism of biological perchlorate reduction, a critical component of the chlorine redox cycle on Earth.

  2. Two fatty acyl reductases involved in moth pheromone biosynthesis.

    PubMed

    Antony, Binu; Ding, Bao-Jian; Moto, Ken'Ichi; Aldosari, Saleh A; Aldawood, Abdulrahman S

    2016-01-01

    Fatty acyl reductases (FARs) constitute an evolutionarily conserved gene family found in all kingdoms of life. Members of the FAR gene family play diverse roles, including seed oil synthesis, insect pheromone biosynthesis, and mammalian wax biosynthesis. In insects, FAR genes dedicated to sex pheromone biosynthesis (pheromone-gland-specific fatty acyl reductase, pgFAR) form a unique clade that exhibits substantial modifications in gene structure and possesses unique specificity and selectivity for fatty acyl substrates. Highly selective and semi-selective 'single pgFARs' produce single and multicomponent pheromone signals in bombycid, pyralid, yponomeutid and noctuid moths. An intriguing question is how a 'single reductase' can direct the synthesis of several fatty alcohols of various chain lengths and isomeric forms. Here, we report two active pgFARs in the pheromone gland of Spodoptera, namely a semi-selective, C14:acyl-specific pgFAR and a highly selective, C16:acyl-specific pgFAR, and demonstrate that these pgFARs play a pivotal role in the formation of species-specific signals, a finding that is strongly supported by functional gene expression data. The study envisages a new area of research for disclosing evolutionary changes associated with C14- and C16-specific FARs in moth pheromone biosynthesis. PMID:27427355

  3. Cloning and Sequence Analysis of Two Pseudomonas Flavoprotein Xenobiotic Reductases

    PubMed Central

    Blehert, David S.; Fox, Brian G.; Chambliss, Glenn H.

    1999-01-01

    The genes encoding flavin mononucleotide-containing oxidoreductases, designated xenobiotic reductases, from Pseudomonas putida II-B and P. fluorescens I-C that removed nitrite from nitroglycerin (NG) by cleavage of the nitroester bond were cloned, sequenced, and characterized. The P. putida gene, xenA, encodes a 39,702-Da monomeric, NAD(P)H-dependent flavoprotein that removes either the terminal or central nitro groups from NG and that reduces 2-cyclohexen-1-one but did not readily reduce 2,4,6-trinitrotoluene (TNT). The P. fluorescens gene, xenB, encodes a 37,441-Da monomeric, NAD(P)H-dependent flavoprotein that exhibits fivefold regioselectivity for removal of the central nitro group from NG and that transforms TNT but did not readily react with 2-cyclohexen-1-one. Heterologous expression of xenA and xenB was demonstrated in Escherichia coli DH5α. The transcription initiation sites of both xenA and xenB were identified by primer extension analysis. BLAST analyses conducted with the P. putida xenA and the P. fluorescens xenB sequences demonstrated that these genes are similar to several other bacterial genes that encode broad-specificity flavoprotein reductases. The prokaryotic flavoprotein reductases described herein likely shared a common ancestor with old yellow enzyme of yeast, a broad-specificity enzyme which may serve a detoxification role in antioxidant defense systems. PMID:10515912

  4. Perchlorate Reductase Is Distinguished by Active Site Aromatic Gate Residues.

    PubMed

    Youngblut, Matthew D; Tsai, Chi-Lin; Clark, Iain C; Carlson, Hans K; Maglaqui, Adrian P; Gau-Pan, Phonchien S; Redford, Steven A; Wong, Alan; Tainer, John A; Coates, John D

    2016-04-22

    Perchlorate is an important ion on both Earth and Mars. Perchlorate reductase (PcrAB), a specialized member of the dimethylsulfoxide reductase superfamily, catalyzes the first step of microbial perchlorate respiration, but little is known about the biochemistry, specificity, structure, and mechanism of PcrAB. Here we characterize the biophysics and phylogeny of this enzyme and report the 1.86-Å resolution PcrAB complex crystal structure. Biochemical analysis revealed a relatively high perchlorate affinity (Km = 6 μm) and a characteristic substrate inhibition compared with the highly similar respiratory nitrate reductase NarGHI, which has a relatively much lower affinity for perchlorate (Km = 1.1 mm) and no substrate inhibition. Structural analysis of oxidized and reduced PcrAB with and without the substrate analog SeO3 (2-) bound to the active site identified key residues in the positively charged and funnel-shaped substrate access tunnel that gated substrate entrance and product release while trapping transiently produced chlorate. The structures suggest gating was associated with shifts of a Phe residue between open and closed conformations plus an Asp residue carboxylate shift between monodentate and bidentate coordination to the active site molybdenum atom. Taken together, structural and mutational analyses of gate residues suggest key roles of these gate residues for substrate entrance and product release. Our combined results provide the first detailed structural insight into the mechanism of biological perchlorate reduction, a critical component of the chlorine redox cycle on Earth. PMID:26940877

  5. An alternative hypothesis for high-T 40Ar/39Ar age spectrum discordance in polyphase extraterrestrial materials

    NASA Astrophysics Data System (ADS)

    Cassata, W. S.; Shuster, D. L.; Renne, P. R.; Weiss, B. P.

    2009-12-01

    A common feature observed in 40Ar/39Ar age spectra of extraterrestrial (ET) rocks is a conspicuous decrease in the ages of high temperature extractions relative to lower temperature steps and a correlated increase in Ca/K, often succeeded by a monotonic increase in ages. This feature is routinely attributed to recoil-implanted 39Ar from a potassium (K)-rich donor phase into a K-poor receptor phase (e.g., 1,2). While 39Ar recoil redistribution is undoubtedly manifested in many terrestrial and ET 40Ar/39Ar whole-rock age spectra, it cannot easily explain the magnitude of high release temperature 40Ar*/39ArK anomalies observed in Martian meteorites ALH 84001 and Nakhla, as well as other course-grained meteorites and lunar rocks. Depending on the aliquot and sample, 50 - 100% of the pyroxene release spectra in ALH 84001 and Nakhla appear strongly perturbed to lower ages. As the mean recoil distance of 39Ar ~0.1 µm, the recoil hypothesis demands that a high-K phase be ubiquitously distributed amongst sub-micron to micron sized pyroxene crystals to account for the observed pyroxene age spectra. However, in both Nakhla and ALH 84001, pyroxene is often completely isolated from high-K phases and individual grains commonly exceed 100 µm in diameter. 40Ar/39Ar analyses of pyroxene-bearing terrestrial basalts, wherein fine-grained pyroxene and plagioclase are intimately adjoined, show that recoil-implanted 39Ar into pyroxene produces much less precipitous anomalies in 40Ar*/39ArK, as predicted by the recoil lengthscale. An alternative hypothesis is that whole-rock age spectra of ET samples with anomalously low ages at high temperatures may reflect diffusive 40Ar distributions within considerably degassed pyroxene grains. Owing to apparent differences in activation energies between glass and/or plagioclase and pyroxene, 40Ar may diffuse more rapidly from pyroxene under certain high-temperature conditions (i.e., above the temperature at which the extrapolated Ar Arrhenius

  6. The 40Ar/39Ar and K/Ar dating of lavas from the Hilo 1-km core hole, Hawaii Scientific Drilling Project

    USGS Publications Warehouse

    Sharp, W.D.; Turrin, B.D.; Renne, P.R.; Lanphere, M.A.

    1996-01-01

    Mauna Kea lava flows cored in the HilIo hole range in age from <200 ka to about 400 ka based on 40Ar/39Ar incremental heating and K-Ar analyses of 16 groundmass samples and one coexisting plagioclase. The lavas, all subaerially deposited, include a lower section consisting only of tholeiitic basalts and an upper section of interbedded alkalic, transitional tholeiitic, and tholeiitic basalts. The lower section has yielded predominantly complex, discordant 40Ar/39Ar age spectra that result from mobility of 40Ar and perhaps K, the presence of excess 40Ar, and redistribution of 39Ar by recoil. Comparison of K-Ar ages with 40Ar/39Ar integrated ages indicates that some of these samples have also lost 39Ar. Nevertheless, two plateau ages of 391 ?? 40 and 400 ?? 26 ka from deep in the hole, combined with data from the upper section, show that the tholeiitic section accumulated at an average rate of about 7 to 8 m/kyr and has an mean recurrence interval of 0.5 kyr/flow unit. Samples from the upper section yield relatively precise 40Ar/39Ar plateau and isotope correlation ages of 326 ?? 23, 241 ?? 5, 232 ?? 4, and 199 ?? 9 ka for depths of -415.7 m to -299.2 m. Within their uncertainty, these ages define a linear relationship with depth, with an average accumulation rate of 0.9 m/kyr and an average recurrence interval of 4.8 kyr/flow unit. The top of the Mauna Kea sequence at -280 m must be older than the plateau age of 132 ?? 32 ka, obtained for the basal Mauna Loa flow in the corehole. The upward decrease in lava accumulation rate is a consequence of the decreasing magma supply available to Mauna Kea as it rode the Pacific plate away from its magma source, the Hawaiian mantle plume. The age-depth relation in the core hole may be used to test and refine models that relate the growth of Mauna Kea to the thermal and compositional structure of the mantle plume.

  7. 40Ar/39Ar geochronology of submarine Mauna Loa volcano, Hawaii

    NASA Astrophysics Data System (ADS)

    Jicha, Brian R.; Rhodes, J. Michael; Singer, Brad S.; Garcia, Michael O.

    2012-09-01

    New geochronologic constraints refine the growth history of Mauna Loa volcano and enhance interpretations of the petrologic, geochemical, and isotopic evolution of Hawaiian magmatism. We report results of 40Ar/39Ar incremental heating experiments on low-K, tholeiitic lavas from the 1.6 km high Kahuku landslide scarp cutting Mauna Loa's submarine southwest rift zone, and from lavas in a deeper section of the rift. Obtaining precise40Ar/39Ar ages from young, tholeiitic lavas containing only 0.2-0.3 wt.% K2O is challenging due to their extremely low radiogenic 40Ar contents. Analyses of groundmass from 45 lavas yield 14 new age determinations (31% success rate) with plateau and isochron ages that agree with stratigraphic constraints. Lavas collected from a 1250 m thick section in the landslide scarp headwall were all erupted around 470 ± 10 ka, implying an extraordinary period of accumulation of ˜25 mm/yr, possibly correlating with the peak of the shield-building stage. This rate is three times higher than the estimated vertical lava accumulation rate for shield-building at Mauna Kea (8.6 ± 3.1 mm/yr) based on results from the Hawaii Scientific Drilling Project. Between ˜470 and 273 ka, the lava accumulation rate along the southwest rift zone decreased dramatically to ˜1 mm/yr. We propose that the marked reduction in lava accumulation rate does not mark the onset of post-shield volcanism as previously suggested, but rather indicates the upward migration of the magma system as Mauna Loa evolved from a submarine stage of growth to one that is predominantly subaerial, thereby cutting off supply to the distal rift zone. Prior to ˜250 ka, lavas with Loihi-like isotopic signatures were erupted along with lavas having typical Mauna Loa values, implying greater heterogeneity in the plume source earlier in Mauna Loa's growth. In addition to refining accumulation rates and the isotopic evolution of the lavas erupted along the southwest rift zone, our new40Ar/39Ar results

  8. 40Ar/39Ar dating of tourmaline as a tool for high-temperature metamorphism thermochronology

    NASA Astrophysics Data System (ADS)

    Jourdan, Fred; Thern, Eric

    2014-05-01

    Tourmaline is an ubiquitous mineral, with properties making it ideal for studying metamorphic processes as well as a useful tool for a wide range of applications (e.g, magmatism, metasomatism, ore deposits [1]), mostly because it is not sensitive to chemical or mechanical alteration and is stable over a wide range of pressure-temperature conditions (up to 6 GPa and 850° C [2]). Typical metamorphic tourmaline types include dravite and shorl which, along with elbaite, belong to the alkali group [1]. The alkali group is notable because tourmalines from this group tend to incorporate trace amounts of K2O and therefore, can be dated using the 40Ar/39Ar technique. In order to understand the maximum temperature below which the K/Ar chronometer stays closed to argon loss by thermally activated diffusion, we carried out temperature controlled furnace diffusion experiments on well-behaved 40Ar/39Ar plateau-forming Archean tourmaline of 2935 ± 9 Ma [3]. Each experiment yielded an Arrhenius profile (Do vs. 1/temperature) that shows that the 39Ar data form two linear arrays with two distinct slopes. The first array only includes a few % of the total gas, has a shallow slope and shows very fast diffusivity at low temperature. We interpret these data as indicating very fast release of argon by cracks and defects. The second array of data points includes most of the gas of each experiment and forms a much steeper slope. These data yielded Ea (activation energy) values ranging from 120 to 157 Kcal/mol and D0 (pre-exponential diffusion factor) values ranging from 1.9x106 to 2.5x109 cm2/s for crystals with an average radius of 100 ± 25 μm. Three additional experiments using a laser (resulting in poor temperature control) suggest similar values although the latter experiments are considered semi-quantitative. The furnace experiments suggest that tourmaline has a weighted mean closure temperature of 804 ± 90 ° C (1σ) for a cooling rate of 10° C/Ma. Monte Carlo simulations using

  9. Inter-monitor standard calibration and tests for Ar-Ar biases

    NASA Astrophysics Data System (ADS)

    Hemming, S. R.; Turrin, B. D.; Swisher, C. C.; Cox, S. E.; Mesko, G. T.; Chang, S.

    2010-12-01

    A major issue facing the geochronology community is that there are biases between chronometers that have become significant as we interrogate the rock record with ever increasing levels of precision. Despite much progress there are still major issues with building a timescale with multiple chronometers and for testing synchroneity of anomalous events in Earth history. Improvements in methods for determining U-Pb zircon dates has led to their application at precisions of 0.2% or better in rocks even younger than a million years (e.g., Crowley et al., 2007, Geology), and significantly better than 0.1% in some cases (e.g., Bowring et al., 2006, Paleontological Society Papers, Volume 12). Additionally, the inter-calibration experiments for U-Pb using the EARTHTIME tracer have yielded excellent agreement among labs (0.05%) and these values are traceable back to SI units through the EARTHTIME tracer calibration experiment (e.g., Condon et al., in press, Geochimica et Cosmochimica Acta). These advances have greatly extended the need for cross calibrations of the two chronometers and ultimately seamless integration into the Geologic Time Scale. The direct comparison of ages using different chronometers and laboratories is the central aspect in the quest for a highly resolved and accurate time scale of Earth History. A significant obstacle to high precision inter-comparison of U-Pb and Ar-Ar age results is the current inability of Ar-Ar labs to achieve agreement on monitor standard ages at the 0.1% level. At the heart of Ar-Ar geochronology is the assumption of a known absolute age of a standard, to which all applicable unknowns are referenced. While individual labs are able to achieve highly precise apparent ages on monitor standards, the lack of a “gold standard” for Ar-Ar dating means that we do not know who is, or indeed if anybody is correct. In order to improve our understanding of factors that may lead to biases in our own laboratories at Lamont-Doherty Earth

  10. The bombardment history of the Moon as recorded by 40Ar-39Ar chronology

    NASA Astrophysics Data System (ADS)

    Fernandes, V. A.; Fritz, J.; Weiss, B. P.; Garrick-Bethell, I.; Shuster, D. L.

    2013-02-01

    New petrography and 40Ar-39Ar ages have been obtained for 1-3 mm sized rock fragments from Apollo 16 Station 13 soil 63503 (North Ray crater ejecta) and chips from three rocks collected by Apollo 16 and Apollo 17 missions. Selection of these samples was aimed at the old 40Ar-39Ar ages to understand the early history of the lunar magnetic field and impact flux. Fifteen samples were studied including crustal material, polymict feldspathic fragmental breccias, and impact melts. The impact ages obtained range between approximately 3.3 and 4.3 billion years (Ga). Polymict fragmental breccia 63503,1 exhibits the lowest signs of recrystallization observed and a probable old relic age of 4.547 ± 0.027. The plateau age of 4.293 ± 0.044 Ga obtained for impact melt rock 63503,13 represents the oldest known age for such a lithology. Possibly, this age represents the minimum age for the South Pole-Aitken (SPA) Basin. In agreement with literature data, these results show that impact ages >3.9 Ga are found in lunar rocks, especially within soil 63503. Impact exhumation of deep-seated warm crustal material onto the lunar surface is considered to explain the common 4.2 Ga ages obtained for weakly shocked samples from soil 63503 and Apollo 17. This would directly imply that one or more basin-forming events occurred at that time. Some rock fragments showing none to limited petrologic features indicate thermal annealing. These rocks may have lost Ar while resident within the hot-ejecta of a large basin. Concurrent with previous studies, these results lead us to advocate for a complex impact flux in the inner solar system during the initial approximately 1.3 Ga.

  11. In Vitro Formation of Nitrate Reductase Using Extracts of the Nitrate Reductase Mutant of Neurospora crassa, nit-1, and Rhodospirillum rubrum

    PubMed Central

    Ketchum, Paul A.; Sevilla, Cynthia L.

    1973-01-01

    In vitro formation of reduced nicotinamide adenine dinucleotide phosphate (NADPH)–nitrate reductase (NADPH: nitrate oxido-reductase, EC 1.6.6.2) has been attained by using extracts of the nitrate reductase mutant of Neurospora crassa, nit-1, and extracts of either photosynthetically or heterotrophically grown Rhodospirillum rubrum, which contribute the constitutive component. The in vitro formation of NADPH-nitrate reductase is characterized by the conversion of the flavin adenine dinucleotide (FAD) stimulated NADPH-cytochrome c reductase, contributed by the N. crassa nit-1 extract from a slower sedimenting form (4.5S) to a faster sedimenting form (7.8S). The 7.8S NADPH-cytochrome c reductase peak coincides in sucrose density gradient profiles with the NADPH–nitrate reductase, FADH2–nitrate reductase and reduced methyl viologen (MVH)–nitrate reductase activities which are also formed in vitro. The constitutive component from R. rubrum is soluble (both in heterotrophically and photosynthetically grown cells), is stimulated by the addition of 10−4 M Na2MoO4 and 10−2 M NaNO3 to cell-free preparations, and has variable activity over the pH range from 3.0 to 9.5. The activity of the constitutive component in some extracts showed a threefold stimulation when the pH was lowered from 6.5 to 4.0. The constitutive activity appears to be associated with a large molecular weight component which sediments as a single peak in sucrose density gradients. However, the constitutive component from R. rubrum is dialyzable and is insensitive to trypsin and protease. These results demonstrate that R. rubrum contains the constitutive component and suggests that it is a low molecular weight, trypsin- and protease-insensitive factor which participates in the in vitro formation of NADPH nitrate reductase. PMID:4270447

  12. Androgen Receptor-Mediated Growth Suppression of HPr-1AR and PC3-Lenti-AR Prostate Epithelial Cells

    PubMed Central

    Bolton, Eric C.

    2015-01-01

    The androgen receptor (AR) mediates the developmental, physiologic, and pathologic effects of androgens including 5α-dihydrotestosterone (DHT). However, the mechanisms whereby AR regulates growth suppression and differentiation of luminal epithelial cells in the prostate gland and proliferation of malignant versions of these cells are not well understood, though they are central to prostate development, homeostasis, and neoplasia. Here, we identify androgen-responsive genes that restrain cell cycle progression and proliferation of human prostate epithelial cell lines (HPr-1AR and PC3-Lenti-AR), and we investigate the mechanisms through which AR regulates their expression. DHT inhibited proliferation of HPr-1AR and PC3-Lenti-AR, and cell cycle analysis revealed a prolonged G1 interval. In the cell cycle, the G1/S-phase transition is initiated by the activity of cyclin D and cyclin-dependent kinase (CDK) complexes, which relieve growth suppression. In HPr-1AR, cyclin D1/2 and CDK4/6 mRNAs were androgen-repressed, whereas CDK inhibitor, CDKN1A, mRNA was androgen-induced. The regulation of these transcripts was AR-dependent, and involved multiple mechanisms. Similar AR-mediated down-regulation of CDK4/6 mRNAs and up-regulation of CDKN1A mRNA occurred in PC3-Lenti-AR. Further, CDK4/6 overexpression suppressed DHT-inhibited cell cycle progression and proliferation of HPr-1AR and PC3-Lenti-AR, whereas CDKN1A overexpression induced cell cycle arrest. We therefore propose that AR-mediated growth suppression of HPr-1AR involves cyclin D1 mRNA decay, transcriptional repression of cyclin D2 and CDK4/6, and transcriptional activation of CDKN1A, which serve to decrease CDK4/6 activity. AR-mediated inhibition of PC3-Lenti-AR proliferation occurs through a similar mechanism, albeit without down-regulation of cyclin D. Our findings provide insight into AR-mediated regulation of prostate epithelial cell proliferation. PMID:26372468

  13. Androgen Receptor-Mediated Growth Suppression of HPr-1AR and PC3-Lenti-AR Prostate Epithelial Cells.

    PubMed

    Kim, Young-Chae; Chen, Congcong; Bolton, Eric C

    2015-01-01

    The androgen receptor (AR) mediates the developmental, physiologic, and pathologic effects of androgens including 5α-dihydrotestosterone (DHT). However, the mechanisms whereby AR regulates growth suppression and differentiation of luminal epithelial cells in the prostate gland and proliferation of malignant versions of these cells are not well understood, though they are central to prostate development, homeostasis, and neoplasia. Here, we identify androgen-responsive genes that restrain cell cycle progression and proliferation of human prostate epithelial cell lines (HPr-1AR and PC3-Lenti-AR), and we investigate the mechanisms through which AR regulates their expression. DHT inhibited proliferation of HPr-1AR and PC3-Lenti-AR, and cell cycle analysis revealed a prolonged G1 interval. In the cell cycle, the G1/S-phase transition is initiated by the activity of cyclin D and cyclin-dependent kinase (CDK) complexes, which relieve growth suppression. In HPr-1AR, cyclin D1/2 and CDK4/6 mRNAs were androgen-repressed, whereas CDK inhibitor, CDKN1A, mRNA was androgen-induced. The regulation of these transcripts was AR-dependent, and involved multiple mechanisms. Similar AR-mediated down-regulation of CDK4/6 mRNAs and up-regulation of CDKN1A mRNA occurred in PC3-Lenti-AR. Further, CDK4/6 overexpression suppressed DHT-inhibited cell cycle progression and proliferation of HPr-1AR and PC3-Lenti-AR, whereas CDKN1A overexpression induced cell cycle arrest. We therefore propose that AR-mediated growth suppression of HPr-1AR involves cyclin D1 mRNA decay, transcriptional repression of cyclin D2 and CDK4/6, and transcriptional activation of CDKN1A, which serve to decrease CDK4/6 activity. AR-mediated inhibition of PC3-Lenti-AR proliferation occurs through a similar mechanism, albeit without down-regulation of cyclin D. Our findings provide insight into AR-mediated regulation of prostate epithelial cell proliferation.

  14. One-electron pseudopotential investigation of the RbAr and FrAr van der Waals systems

    NASA Astrophysics Data System (ADS)

    Dhiflaoui, J.; Berriche, H.

    2012-12-01

    The potential energy curves of the ground state and many excited states of RbAr and FrAr van der Waals systems have been determined using a one-electron pseudopotential approach. The pseudopotential technique is used to replace the effect of the Rb+ and Fr+ cores and the electron-Ar interaction. In addition a core-core interaction is included. This has permitted to reduce the number of active electrons of the RbAr and FrAr systems to only one electron, the valence electron. This has led to use very large basis sets for Rb, Fr and Ar atoms. In this context, the potential energy curves of the ground and many excited states are performed at the SCF level. The core-core interactions for Rb+Ar and Fr+Ar are included using the CCSD(T) accurate potentials of Hickling et al. [H. Hickling, L. Viehland, D. Shepherd, P. Soldan, E. Lee and T. Wright, Phys. Chem. Chem. Phys. 6 (2004) 4233]. In addition, the spectroscopic constants of these states are derived and compared with the available theoretical works. Such comparison for RbAr has shown a very good agreement for the ground and the first excited states. However, the FrAr system was not studied previously and its spectroscopic constants are presented here for the first time.

  15. Selenite reduction by Shewanella oneidensis MR-1 is mediated by fumarate reductase in periplasm

    NASA Astrophysics Data System (ADS)

    Li, Dao-Bo; Cheng, Yuan-Yuan; Wu, Chao; Li, Wen-Wei; Li, Na; Yang, Zong-Chuang; Tong, Zhong-Hua; Yu, Han-Qing

    2014-01-01

    In situ reduction of selenite to elemental selenium (Se(0)), by microorganisms in sediments and soils is an important process and greatly affects the environmental distribution and the biological effects of selenium. However, the mechanism behind such a biological process remains unrevealed yet. Here we use Shewanella oneidensis MR-1, a widely-distributed dissimilatory metal-reducing bacterium with a powerful and diverse respiration capability, to evaluate the involvement of anaerobic respiration system in the microbial selenite reduction. With mutants analysis, we identify fumarate reductase FccA as the terminal reductase of selenite in periplasm. Moreover, we find that such a reduction is dependent on central respiration c-type cytochrome CymA. In contrast, nitrate reductase, nitrite reductase, and the Mtr electron transfer pathway do not work as selenite reductases. These findings reveal a previously unrecognized role of anaerobic respiration reductases of S. oneidensis MR-1 in selenite reduction and geochemical cycles of selenium in sediments and soils.

  16. Triaxial superdeformation in {sup 40}Ar

    SciTech Connect

    Taniguchi, Yasutaka; Ikeda, Kiyomi; Kanada-En'yo, Yoshiko; Kimura, Masaaki; Horiuchi, Hisashi; Ideguchi, Eiji

    2010-07-15

    Superdeformed (SD) states in {sup 40}Ar have been studied using the deformed-basis antisymmetrized molecular dynamics. Low-energy states were calculated by the parity and angular momentum projection and the generator coordinate method (GCM). Basis wave functions were obtained by the energy variation with a constraint on the quadrupole deformation parameter beta, while other quantities such as triaxiality gamma were optimized by the energy variation. By the GCM calculation, an SD band was obtained just above the ground-state band. The SD band involves a K{sup p}i=2{sup +} side band due to the triaxiality. The calculated electric quadrupole transition strengths of the SD band reproduce the experimental values appropriately. Triaxiality is significant for understanding low-lying states.

  17. High resolution photoionisation spectroscopy of vibrationally excited Ar · NO

    NASA Astrophysics Data System (ADS)

    Monti, O. L. A.; Cruse, H. A.; Softley, T. P.; Mackenzie, S. R.

    2001-01-01

    Mass-analysed threshold ionisation (MATI) spectra of the Ar · NO complex have been obtained for the first time. These spectra have been used to determine unambiguously the nature of three bands detected by resonance-enhanced multiphoton ionisation (REMPI) spectroscopy via the à state of Ar · NO. The features are shown to originate from vibrationally excited states of Ar · NO in its electronic ground state. The assignment is in agreement with recent theoretical calculations.

  18. Cross sections of the 36Ar(d,α)34mCl, 40Ar(d,α)38Cl, and 40Ar(d,p)41Ar nuclear reactions below 8.4 MeV.

    PubMed

    Engle, J W; Severin, G W; Barnhart, T E; Knutson, L D; Nickles, R J

    2012-02-01

    We have measured the cross section for production of the medically interesting isotope (34m)Cl, along with (38)Cl and (41)Ar, using deuteron bombardments of (36)Ar and (40)Ar below 8.4 MeV. ALICE/ASH analytical codes were employed to determine the shape of nuclear excitation functions, and experiments were performed using the University of Wisconsin tandem electrostatic accelerator to irradiate thin targets of argon gas.

  19. Plasma AR and abiraterone-resistant prostate cancer.

    PubMed

    Romanel, Alessandro; Gasi Tandefelt, Delila; Conteduca, Vincenza; Jayaram, Anuradha; Casiraghi, Nicola; Wetterskog, Daniel; Salvi, Samanta; Amadori, Dino; Zafeiriou, Zafeiris; Rescigno, Pasquale; Bianchini, Diletta; Gurioli, Giorgia; Casadio, Valentina; Carreira, Suzanne; Goodall, Jane; Wingate, Anna; Ferraldeschi, Roberta; Tunariu, Nina; Flohr, Penny; De Giorgi, Ugo; de Bono, Johann S; Demichelis, Francesca; Attard, Gerhardt

    2015-11-01

    Androgen receptor (AR) gene aberrations are rare in prostate cancer before primary hormone treatment but emerge with castration resistance. To determine AR gene status using a minimally invasive assay that could have broad clinical utility, we developed a targeted next-generation sequencing approach amenable to plasma DNA, covering all AR coding bases and genomic regions that are highly informative in prostate cancer. We sequenced 274 plasma samples from 97 castration-resistant prostate cancer patients treated with abiraterone at two institutions. We controlled for normal DNA in patients' circulation and detected a sufficiently high tumor DNA fraction to quantify AR copy number state in 217 samples (80 patients). Detection of AR copy number gain and point mutations in plasma were inversely correlated, supported further by the enrichment of nonsynonymous versus synonymous mutations in AR copy number normal as opposed to AR gain samples. Whereas AR copy number was unchanged from before treatment to progression and no mutant AR alleles showed signal for acquired gain, we observed emergence of T878A or L702H AR amino acid changes in 13% of tumors at progression on abiraterone. Patients with AR gain or T878A or L702H before abiraterone (45%) were 4.9 and 7.8 times less likely to have a ≥50 or ≥90% decline in prostate-specific antigen (PSA), respectively, and had a significantly worse overall [hazard ratio (HR), 7.33; 95% confidence interval (CI), 3.51 to 15.34; P = 1.3 × 10(-9)) and progression-free (HR, 3.73; 95% CI, 2.17 to 6.41; P = 5.6 × 10(-7)) survival. Evaluation of plasma AR by next-generation sequencing could identify cancers with primary resistance to abiraterone.

  20. Unmixing 40Ar/39Ar Muscovite Ages Using Powder X-ray Diffraction

    NASA Astrophysics Data System (ADS)

    McAleer, R. J.; Kunk, M. J.; Valley, P. M.; Walsh, G. J.; Bish, D. L.; Wintsch, R. P.

    2014-12-01

    Whole rock powder X-ray diffraction (XRD) experiments from eight samples collected across a retrograde ductile shear zone in the Devonian Littleton Formation near Claremont, NH, exhibit broad and asymmetric to bimodal muscovite 00l reflections. These composite 00l reflections exhibit a systematic change in shape with increasing retrograde strain. Microtextural relationships, electron microprobe quantitative analyses, and element mapping indicate that the change in peak shape reflects progressive dissolution of metastable Na-rich muscovite and the precipitation of stable Na-poor muscovite. 40Ar/39Ar step heating experiments on muscovite concentrates from these samples show a decrease in total gas age from 274 to 258 Ma as the highest strain zone is approached, and steps within individual spectra range in age by ~20 m.y. The correlation between age and 00l peak shape suggests that the argon isotopic system also tracks the dissolution-precipitation process. Furthermore, the variation in age during step heating indicates that these populations exhibit different in-vacuo degassing behavior. Comparison of whole rock and muscovite concentrate XRD patterns from the same samples shows that the mineral separation process can fractionate these muscovite populations. With this knowledge, four muscovite concentrates were prepared from a single hand sample, analyzed by XRD, and dated. Combining modal estimates from XRD experiments with total gas ages, the four splits narrowly define a mixing line that resolves end-member ages of 250 and 300 Ma for the neocrystallized and earlier high grade populations of muscovite, respectively. These ages are consistent with age data from all other samples. The results show that, in some settings, powder XRD provides a powerful and time effective method to both identify the existence of and establish the proportions of multiple compositional populations of muscovite prior to 40Ar/39Ar analysis. This approach will be especially useful in

  1. Statistical estimation of mineral age by K-Ar method

    SciTech Connect

    Vistelius, A.B.; Drubetzkoy, E.R.; Faas, A.V. )

    1989-11-01

    Statistical estimation of age of {sup 40}Ar/{sup 40}K ratios may be considered a result of convolution of uniform and normal distributions with different weights for different minerals. Data from Gul'shad Massif (Nearbalkhash, Kazakhstan, USSR) indicate that {sup 40}Ar/{sup 40}K ratios reflecting the intensity of geochemical processes can be resolved using convolutions. Loss of {sup 40}Ar in biotites is shown whereas hornblende retained the original content of {sup 40}Ar throughout the geological history of the massif. Results demonstrate that different estimation methods must be used for different minerals and different rocks when radiometric ages are employed for dating.

  2. β -delayed three-proton decay of 31Ar

    NASA Astrophysics Data System (ADS)

    Lis, A. A.; Mazzocchi, C.; Dominik, W.; Janas, Z.; Pfützner, M.; Pomorski, M.; Acosta, L.; Baraeva, S.; Casarejos, E.; Duénas-Díaz, J.; Dunin, V.; Espino, J. M.; Estrade, A.; Farinon, F.; Fomichev, A.; Geissel, H.; Gorshkov, A.; Kamiński, G.; Kiselev, O.; Knöbel, R.; Krupko, S.; Kuich, M.; Litvinov, Yu. A.; Marquinez-Durán, G.; Martel, I.; Mukha, I.; Nociforo, C.; Ordúz, A. K.; Pietri, S.; Prochazka, A.; Sánchez-Benítez, A. M.; Simon, H.; Sitar, B.; Slepnev, R.; Stanoiu, M.; Strmen, P.; Szarka, I.; Takechi, M.; Tanaka, Y.; Weick, H.; Winfield, J. S.

    2015-06-01

    The β decay of 31Ar , produced by fragmentation of an 36Ar beam at 880 MeV/nucleon, was investigated. Identified ions of 31Ar were stopped in a gaseous time projection chamber with optical readout allowing us to record decay events with emission of protons. In addition to β -delayed emission of one and two protons we clearly observed the β -delayed three-proton branch. The branching ratio for this channel in 31Ar is found to be 0.07 ±0.02 %.

  3. Age and Duration of Weathering by 40K-40Ar and 40Ar/39Ar Analysis of Potassium-Manganese Oxides.

    PubMed

    Vasconcelos, P M; Becker, T A; Renne, P R; Brimhall, G H

    1992-10-16

    Supergene cryptomelane [K(1-2)(Mn(3+)Mn(4+))(8)O(16). chiH(2)O] samples from deeply weathered pegmatites in southeastern Brazil subjected to (40)K-(40)Ar and (40)Ar/(39)Ar analysis yielded (40)K-(40)Ar dates ranging from 10.1 +/- 0.5 to 5.6 +/- 0.2 Ma (million years ago). Laser-probe (40)Ar/(39)Ar step-heating of the two most disparate samples yielded plateau dates of 9.94 +/- 0.05 and 5.59 +/- 0.10 Ma, corresponding, within 2 sigma, to the (40)K-(40)Ar dates. The results imply that deep weathering profiles along the eastern Brazilian margin do not reflect present climatic conditions but are the result of a long-term process that was already advanced by the late Miocene. Weathering ages predate pulses of continental sedimentation along the eastern Brazilian margin and suggest that there was a time lag between weathering and erosion processes and sedimentation processes.

  4. 40Ar/39Ar ages from the rhyolite of Alder Creek, California: age of the Cobb Mountain normal-polarity subchron revisited

    USGS Publications Warehouse

    Turrin, B.D.; Donnelly-Nolan, J. M.; Hearn, B.C., Jr.

    1994-01-01

    New 40Ar/39Ar age determinations on sanidine from the rhyolite of Alder Creek, California, indicate a 1.186 ?? 0.006 Ma age for the Cobb Mountain Normal-Polarity Subchron. The hew age is statistically older (?? = 0.05) than the previously reported K-Ar age (1.12 ?? 0.02 Ma) and agrees with the age suggested by the astronomical polarity time scale. Incomplete extraction of radiogenic 40Ar (40Ar*) from the sanidine is the most likely reason for the disparity between the 40Ar/39Ar and K-Ar ages. Because the Cobb Mountain subchron is a worldwide, short-duration event, and because no widely used interlaboratory 40Ar/39Ar standard younger than 27 Ma exists, it is proposed that sanidine from the rhyolite of Alder Creek be considered for use as a new Quaternary 40Ar/39Ar mineral standard. -Authors

  5. Ar-39 - Ar-40 Evidence for an Approximately 4.26 Ga Impact Heating Event on the LL Parent Body

    NASA Technical Reports Server (NTRS)

    Dixon, E. T.; Bogard, D. D.; Rubin, A. E.

    2003-01-01

    Miller Range 99301 is a type 6, unbrecciated LL chondrite. MIL 99301 is of interest because some compositional and petrographic features suggest it experienced rather high shock grades, whereas other features suggest it is relatively unshocked. Inconsistent shock indicators could be explained if MIL 99301 was shocked but then partly annealed by heat produced by impacts on the parent body. The hypothesis that MIL 99301 experienced high temperature metamorphism (type 6) followed by a later shock event that heated, but did not melt, the constituent feldspar can be evaluated using (39)Ar-(40)Ar chronology. This is because (39)Ar-(40)Ar ages of shocked ordinary chondrites are generally <4.2 Ga, whereas (39)Ar-(40)Ar ages of unshocked meteorites are generally older, and between 4.52 - 4.38 Ga.

  6. The mechanism of the quinone reductase reaction of pig heart lipoamide dehydrogenase.

    PubMed Central

    Vienozinskis, J; Butkus, A; Cenas, N; Kulys, J

    1990-01-01

    The relationship between the NADH:lipoamide reductase and NADH:quinone reductase reactions of pig heart lipoamide dehydrogenase (EC 1.6.4.3) was investigated. At pH 7.0 the catalytic constant of the quinone reductase reaction (kcat.) is 70 s-1 and the rate constant of the active-centre reduction by NADH (kcat./Km) is 9.2 x 10(5) M-1.s-1. These constants are almost an order lower than those for the lipoamide reductase reaction. The maximal quinone reductase activity is observed at pH 6.0-5.5. The use of [4(S)-2H]NADH as substrate decreases kcat./Km for the lipoamide reductase reaction and both kcat. and kcat./Km for the quinone reductase reaction. The kcat./Km values for quinones in this case are decreased 1.85-3.0-fold. NAD+ is a more effective inhibitor in the quinone reductase reaction than in the lipoamide reductase reaction. The pattern of inhibition reflects the shift of the reaction equilibrium. Various forms of the four-electron-reduced enzyme are believed to reduce quinones. Simple and 'hybrid ping-pong' mechanisms of this reaction are discussed. The logarithms of kcat./Km for quinones are hyperbolically dependent on their single-electron reduction potentials (E1(7]. A three-step mechanism for a mixed one-electron and two-electron reduction of quinones by lipoamide dehydrogenase is proposed. PMID:2375745

  7. Selenate reductase activity in Escherichia coli requires Isc iron-sulfur cluster biosynthesis genes.

    PubMed

    Yee, Nathan; Choi, Jessica; Porter, Abigail W; Carey, Sean; Rauschenbach, Ines; Harel, Arye

    2014-12-01

    The selenate reductase in Escherichia coli is a multi-subunit enzyme predicted to bind Fe-S clusters. In this study, we examined the iron-sulfur cluster biosynthesis genes that are required for selenate reductase activity. Mutants devoid of either the iscU or hscB gene in the Isc iron-sulfur cluster biosynthesis pathway lost the ability to reduce selenate. Genetic complementation by the wild-type sequences restored selenate reductase activity. The results indicate the Isc biosynthetic system plays a key role in selenate reductase Fe-S cofactor assembly and is essential for enzyme activity.

  8. Identification of the 7-Hydroxymethyl Chlorophyll a Reductase of the Chlorophyll Cycle in Arabidopsis[W

    PubMed Central

    Meguro, Miki; Ito, Hisashi; Takabayashi, Atsushi; Tanaka, Ryouichi; Tanaka, Ayumi

    2011-01-01

    The interconversion of chlorophyll a and chlorophyll b, referred to as the chlorophyll cycle, plays a crucial role in the processes of greening, acclimation to light intensity, and senescence. The chlorophyll cycle consists of three reactions: the conversions of chlorophyll a to chlorophyll b by chlorophyllide a oxygenase, chlorophyll b to 7-hydroxymethyl chlorophyll a by chlorophyll b reductase, and 7-hydroxymethyl chlorophyll a to chlorophyll a by 7-hydroxymethyl chlorophyll a reductase. We identified 7-hydroxymethyl chlorophyll a reductase, which is the last remaining unidentified enzyme of the chlorophyll cycle, from Arabidopsis thaliana by genetic and biochemical methods. Recombinant 7-hydroxymethyl chlorophyll a reductase converted 7-hydroxymethyl chlorophyll a to chlorophyll a using ferredoxin. Both sequence and biochemical analyses showed that 7-hydroxymethyl chlorophyll a reductase contains flavin adenine dinucleotide and an iron-sulfur center. In addition, a phylogenetic analysis elucidated the evolution of 7-hydroxymethyl chlorophyll a reductase from divinyl chlorophyllide vinyl reductase. A mutant lacking 7-hydroxymethyl chlorophyll a reductase was found to accumulate 7-hydroxymethyl chlorophyll a and pheophorbide a. Furthermore, this accumulation of pheophorbide a in the mutant was rescued by the inactivation of the chlorophyll b reductase gene. The downregulation of pheophorbide a oxygenase activity is discussed in relation to 7-hydroxymethyl chlorophyll a accumulation. PMID:21934147

  9. Differential unroofing within the central metasedimentary Belt of the Grenville Orogen: constraints from 40Ar/39Ar thermochronology

    USGS Publications Warehouse

    Cosca, M.A.; Essene, E.J.; Kunk, M.J.; Sutter, J.F.

    1992-01-01

    An 40Ar/39Ar thermochronological investigation of upper greenschist to granulite facies gneiss, amphibolite and marble was conducted in the Central Metasedimentary Belt (CMB), Ontario, to constrain its cooling history. Incremental 40Ar/39Ar release spectra indicate that substantial differential unroofing occurred in the CMB between ??? 1000 and ??? 600 Ma. A consistent pattern of significantly older hornblende and phlogopite 40Ar/3Ar cooling ages on the southeast sides of major northeast striking shear zones is interpreted to reflect late displacement due to extensional deformation. Variations in hornblende 40Ar/39Ar age plateaus exceeding 200 Ma occur over distances less than 50 km with major age discontinuities occurring across the Robertson Lake shear zone and the Sharbot Lake mylonite zone which separate the Sharbot Lake terrane from the Elzevir and Frontenac terranes. Extensional displacements of up to 14 km are inferred between the Frontenac and Elzevir terranes of the CMB. No evidence for significant post argon-closure vertical displacement is indicated in the vicinity of the Perth Road mylonite within the Frontenac terrane. Variations of nearly 100 Ma in phlogopite 40Ar/39Ar plateau ages occur in undeformed marble on either side of the Bancroft Shear Zone. Phlogopites from sheared and mylonitized marble within the shear zone yield 40Ar/39Ar diffusional loss profiles, but have older geologically meaningless ages thought to reflect incorporation of excess argon. By ??? 900 Ma, southeast directed extension was occurring throughout the CMB, possibly initiated along previous zones of compressional shearing. An easterly migration of active zones of extension is inferred, possibly related to an earlier, overall easterly migration of active zones of regional thrusting and easterly migration of an ancient subduction zone. The duration of extensional shearing is not well constrained, but must have ceased before ??? 600 Ma as required by the deposition of overlying

  10. 40Ar/39Ar Age of the Lathrop Wells Volcanic Center, Yucca Mountain, Nevada.

    PubMed

    Turrin, B D; Champion, D; Fleck, R J

    1991-08-01

    Paleomagnetic and (40)Ar/(39)Ar analyses from the Lathrop Wells volcanic center, Nevada, indicate that two eruptive events have occurred there. The ages (136 +/- 8 and 141 +/- 9 thousand years ago) for these two events are analytically indistinguishable. The small angular difference (4.7 degrees ) between the paleomagnetic directions from these two events suggests they differ in age by only about 100 years. These ages are consistent with the chronology of the surficial geological units in the Yucca Mountain area. These results contradict earlier interpretations of the cinder-cone geomorphology and soil-profile data that suggest that at least five temporally discrete eruptive events occurred at Lathrop Wells approximately 20,000 years ago. PMID:17772371

  11. Acoustic resonance spectroscopy (ARS): ARS300 operations manual, software version 2.01

    SciTech Connect

    1996-07-25

    Acoustic Resonance Spectroscopy (ARS) is a nondestructive evaluation technology developed at the Los Alamos National Laboratory. The ARS technique is a fast, safe, and nonintrusive technique that is particularly useful when a large number of objects need to be tested. Any physical object, whether solid, hollow, or fluid filled, has many modes of vibration. These modes of vibration, commonly referred to as the natural resonant modes or resonant frequencies, are determined by the object`s shape, size, and physical properties, such as elastic moduli, speed of sound, and density. If the object is mechanically excited at frequencies corresponding to its characteristic natural vibrational modes, a resonance effect can be observed when small excitation energies produce large amplitude vibrations in the object. At other excitation frequencies, i.e., vibrational response of the object is minimal.

  12. 40Ar/39Ar Age of the Lathrop Wells Volcanic Center, Yucca Mountain, Nevada.

    PubMed

    Turrin, B D; Champion, D; Fleck, R J

    1991-08-01

    Paleomagnetic and (40)Ar/(39)Ar analyses from the Lathrop Wells volcanic center, Nevada, indicate that two eruptive events have occurred there. The ages (136 +/- 8 and 141 +/- 9 thousand years ago) for these two events are analytically indistinguishable. The small angular difference (4.7 degrees ) between the paleomagnetic directions from these two events suggests they differ in age by only about 100 years. These ages are consistent with the chronology of the surficial geological units in the Yucca Mountain area. These results contradict earlier interpretations of the cinder-cone geomorphology and soil-profile data that suggest that at least five temporally discrete eruptive events occurred at Lathrop Wells approximately 20,000 years ago.

  13. 40Ar/39Ar age of the Lathrop Wells volcanic center, Yucca Mountain, Nevada

    USGS Publications Warehouse

    Turrin, B.D.; Champion, D.; Fleck, R.J.

    1991-01-01

    Paleomagnetic and 40Ar/39Ar analyses from the Lathrop Wells volcanic center, Nevada, indicate that two eruptive events have occurred there. The ages (136 ?? 8 and 141 ?? 9 thousand years ago) for these two events are analytically indistinguishable. The small angular difference (4.7??) between the paleomagnetic directions from these two events suggests they differ in age by only about 100 years. These ages are consistent with the chronology of the surficial geological units in the Yucca Mountain area. These results contradict earlier interpretations of the cinder-cone geomorphology and soil-profile data that suggest that at least five temporally discrete eruptive events occurred at Lathrop Wells approximately 20,000 years ago.

  14. 40Ar/39Ar dates from the West Siberian Basin: Siberian flood basalt province doubled.

    PubMed

    Reichow, Marc K; Saunders, Andrew D; White, Rosalind V; Pringle, Malcolm S; Al'Mukhamedov, Alexander I; Medvedev, Alexander I; Kirda, Nikolay P

    2002-06-01

    Widespread basaltic volcanism occurred in the region of the West Siberian Basin in central Russia during Permo-Triassic times. New 40Ar/39Ar age determinations on plagioclase grains from deep boreholes in the basin reveal that the basalts were erupted 249.4 +/- 0.5 million years ago. This is synchronous with the bulk of the Siberian Traps, erupted further east on the Siberian Platform. The age and geochemical data confirm that the West Siberian Basin basalts are part of the Siberian Traps and at least double the confirmed area of the volcanic province as a whole. The larger area of volcanism strengthens the link between the volcanism and the end-Permian mass extinction. PMID:12052954

  15. In-situ Ar isotope, 40Ar/39Ar analysis and mineral chemistry of nosean in the phonolite from Olbrück volcano, East Eifel volcanic field, Germany: Implication for the source of excess 40Ar

    NASA Astrophysics Data System (ADS)

    Sudo, Masafumi; Altenberger, Uwe; Günter, Christina

    2014-05-01

    Since the report by Lippolt et al. (1990), hauyne and nosean phenocrysts in certain phonolites from the northwest in the Quaternary East Eifel volcanic field in Germany were known to contain significant amounts of excess 40Ar, thus, show apparent older ages than the other minerals. However, its petrographic meaning have not been well known. Meanwhile, Sumino et al. (2008) has identified the source of the excess 40Ar in the plagioclase phenocrysts from the historic Unzen dacite lava as the melt inclusions in the zones parallely developed to the plagioclase rim by in-situ laser Ar isotope analysis. In order to obtain eruption ages of very young volcanoes as like Quaternary Eifel volcanic field by the K-Ar system, it is quite essential to know about the location of excess 40Ar in volcanic rocks. We have collected phonolites from the Olbrück volcano in East Eifel and investigated its petrography and mineral chemistry and also performed in-situ Ar isotope analyses of unirradiated rock section sample and also in-situ 40Ar/39Ar analysis of neutron irradiated section sample with the UV pulse laser (wavelength 266 nm) and 40Ar/39Ar analytical system of the University of Potsdam. Petrographically, nosean contained fine melt and/or gas inclusions of less than 5 micrometer, which mostly distribute linearly and are relatively enriched in chlorine than the areas without inclusions. Solid inclusions of similar sizes contain CaO and fluorine. In nosean, typically around 5 wt% of sulfur is contained. The 40Ar/39Ar dating was also performed to leucite, sanidine and groundmass in the same section for comparison of those ages with that of nosean. In each analysis, 200 micrometer of beam size was used for making a pit with depth of up to 300 micrometer by laser ablation. As our 40Ar/39Ar analyses were conducted one and half year after the neutron irradiation, thus, short lived 37Ar derived from Ca had decayed very much, we measured Ca and K contents in nosean by SEM-EDS then applied

  16. The Berkeley Instrumental Neutron Generator (BINGE) for 40Ar/39Ar geochronology

    NASA Astrophysics Data System (ADS)

    Renne, P. R.; Becker, T. A.; Bernstein, L.; Firestone, R. B.; Kirsch, L.; Leung, K. N.; Rogers, A.; Van Bibber, K.; Waltz, C.

    2014-12-01

    The Berkeley Instrumental Neutron Generator (BINGE) facility is the product of a consortium involving the Berkeley Geochronology Center (BGC), the U.C. Berkeley Nuclear Engineering Dept. (UCB/NE), and Lawrence Berkeley (LBNL) and Lawrence Livermore (LLNL) National Labs. BINGE was initially designed (and funded by NSF) for 40Ar/39Ar geochronology. BINGE uses a plasma-based deuteron ion source and a self-loading Ti-surfaced target to induce deuteron-deuterium (DD) fusion via the reaction 2H(d,n)3He, producing 2.45 MeV neutrons. The limited neutron energy spectrum is aimed at reducing recoil effects, interfering nuclear reactions, and unwanted radioactive byproducts, all of which are undesirable consequences of conventional irradiation with 235U fission spectrum neutrons. Minimization of interfering reactions such as 40Ca(n,na)36Ar greatly reduces penalties for over-irradiation, enabling improved signal/background measurement of e.g. 39Ar. BINGE will also be used for a variety of nuclear physics and engineering experiments that require a high flux of monoenergetic neutrons. Neutron energies lower than 2.45 MeV can be obtained via irradiation ports within and external to polyethylene shielding. Initial commissioning produced a neutron flux of 108 n/sec/cm2 at 1 mA source current and 100 kV anode voltage, as expected. When scaled up to the 1 A source current as planned, this indicates that BINGE will achieve the design objective neutron flux of 1011 n/sec/cm2. Further progress towards this goal will be reported. Supported by NSF (grant #EAR-0960138), BGC, UCB/NE, University of California Office of the President, and DOE through LLNL under contract #DE-AC52-07NA27344 and LBNL under contract #DE-AC02-05CH11231.

  17. Refining lunar impact chronology through high spatial resolution 40Ar/39Ar dating of impact melts

    PubMed Central

    Mercer, Cameron M.; Young, Kelsey E.; Weirich, John R.; Hodges, Kip V.; Jolliff, Bradley L.; Wartho, Jo-Anne; van Soest, Matthijs C.

    2015-01-01

    Quantitative constraints on the ages of melt-forming impact events on the Moon are based primarily on isotope geochronology of returned samples. However, interpreting the results of such studies can often be difficult because the provenance region of any sample returned from the lunar surface may have experienced multiple impact events over the course of billions of years of bombardment. We illustrate this problem with new laser microprobe 40Ar/39Ar data for two Apollo 17 impact melt breccias. Whereas one sample yields a straightforward result, indicating a single melt-forming event at ca. 3.83 Ga, data from the other sample document multiple impact melt–forming events between ca. 3.81 Ga and at least as young as ca. 3.27 Ga. Notably, published zircon U/Pb data indicate the existence of even older melt products in the same sample. The revelation of multiple impact events through 40Ar/39Ar geochronology is likely not to have been possible using standard incremental heating methods alone, demonstrating the complementarity of the laser microprobe technique. Evidence for 3.83 Ga to 3.81 Ga melt components in these samples reinforces emerging interpretations that Apollo 17 impact breccia samples include a significant component of ejecta from the Imbrium basin impact. Collectively, our results underscore the need to quantitatively resolve the ages of different melt generations from multiple samples to improve our current understanding of the lunar impact record, and to establish the absolute ages of important impact structures encountered during future exploration missions in the inner Solar System. PMID:26601128

  18. Ar-40/Ar-39 Age of Hornblende-bearing R Chondrite LAP 04840

    NASA Technical Reports Server (NTRS)

    Righter, K.; Cosca, M.

    2014-01-01

    Chondrites have a complex chronology due to several variables affecting and operating on chondritic parent bodies such as radiogenic heating, pressure and temperature variation with depth, aqueous alteration, and shock or impact heating [1]. Unbrecciated chondrites can record ages from 4.56 to 4.4 Ga that represent cooling in small parent bodies. Some brecciated chondrites exhibit younger ages (<<4 to 4.4 Ga) that may reflect the age of brecciation, disturbance, or shock and impact events (<< 4 Ga). A unique R chondrite was recently found in the LaPaz Icefield of Antarctica - LAP 04840 [2]. This chondrite contains approx.15% hornblende and trace amounts of biotite, making it the first of its kind. Studies have revealed an equigranular texture, mineral equilibria yielding equilibration near 650-700 C and 250-500 bars, hornblende that is dominantly OH-bearing (very little Cl or F), and high D/H ratios [8,9,10]. To help gain a better understanding of the origin of this unique sample, we have measured the Ar-40/Ar-39 age. Age of 4.290 +/- 0.030 Ga is younger than one would expect for a sample that has cooled within a small body [4], and one might instead attribute the age to a younger shock event, On the other hand, there is no evidence for extensive shock in this meteorite (shock stage S2; [3]), so this sample may have been reannealed after the shock event. This age is similar to Ar-Ar ages determined for some other R chondrites

  19. 40Ar/39Ar geochronology and paleomagnetism of Independence volcano, Absaroka volcanic supergroup, Beartooth mountains, Montana

    USGS Publications Warehouse

    Harlan, S.S.; Snee, L.W.; Geissman, J.W.

    1996-01-01

    Independence volcano is a major volcanic complex in the lower part of the Absaroka Volcanic Supergroup (AVS) of Montana and Wyoming. Recently reported Rb-Sr mineral dates from the complex give apparent ages of 91 and 84 Ma, whereas field relationships and the physical and compositional similarity of the rocks with other dated parts of the AVS indicate an Early to Middle Eocene age for eruption and deposition. To resolve the conflict between age assignments based on stratigraphic correlations and Rb-Sr dates, we report new paleomagnetic data and 40Ar/39Ar dates for Independence volcano. Paleomagnetic data for the stock and an and andesite plug that cuts the stock are well grouped, of reverse polarity, and yield a virtual geomagnetic pole that is essentially identical to Late Cretaceous and Tertiary reference poles. The reverse polarity indicates that the magnetization of these rocks is probably younger than the Cretaceous normal superchron, or less than about 83.5 Ma. Hornblende from a volcanic breccia near the base of the volcanic pile gives a 40Ar/39Ar age of 51.57 Ma, whereas biotites from a dacite sill and a granodiorite stock that forms the core of the volcano give dates that range from 49.96 to 48.50 Ma. These dates record the age of eruption and intrusion of these rocks and clearly show that the age of Independence volcano is Early to Middle Eocene, consistent with stratigraphic relations. We suggest that the Rb-Sr mineral dates from the Independence stock and related intrusions are unreliable.

  20. Refining lunar impact chronology through high spatial resolution (40)Ar/(39)Ar dating of impact melts.

    PubMed

    Mercer, Cameron M; Young, Kelsey E; Weirich, John R; Hodges, Kip V; Jolliff, Bradley L; Wartho, Jo-Anne; van Soest, Matthijs C

    2015-02-01

    Quantitative constraints on the ages of melt-forming impact events on the Moon are based primarily on isotope geochronology of returned samples. However, interpreting the results of such studies can often be difficult because the provenance region of any sample returned from the lunar surface may have experienced multiple impact events over the course of billions of years of bombardment. We illustrate this problem with new laser microprobe (40)Ar/(39)Ar data for two Apollo 17 impact melt breccias. Whereas one sample yields a straightforward result, indicating a single melt-forming event at ca. 3.83 Ga, data from the other sample document multiple impact melt-forming events between ca. 3.81 Ga and at least as young as ca. 3.27 Ga. Notably, published zircon U/Pb data indicate the existence of even older melt products in the same sample. The revelation of multiple impact events through (40)Ar/(39)Ar geochronology is likely not to have been possible using standard incremental heating methods alone, demonstrating the complementarity of the laser microprobe technique. Evidence for 3.83 Ga to 3.81 Ga melt components in these samples reinforces emerging interpretations that Apollo 17 impact breccia samples include a significant component of ejecta from the Imbrium basin impact. Collectively, our results underscore the need to quantitatively resolve the ages of different melt generations from multiple samples to improve our current understanding of the lunar impact record, and to establish the absolute ages of important impact structures encountered during future exploration missions in the inner Solar System. PMID:26601128

  1. 40Ar/39Ar thermochronology of mesoproterozoic metamorphism in the Colorado Front Range

    USGS Publications Warehouse

    Shaw, C.A.; Snee, L.W.; Selverstone, J.; Reed, J.C.

    1999-01-01

    A low-pressure metamorphic episode in the Colorado Front Range has been identified by the presence of staurolite, andalusite, cordierite, and garnet porphyroblasts overprinting earlier assemblages. The overprinting assemblages and reaction textures are most consistent with porphyroblast growth on a prograde metamorphic path with peak temperatures exceeding ~525??C. Twenty-eight 40Ar/39Ar dates on hornblende, muscovite, biotite, and microcline were used to infer the age and thermal conditions of metamorphism. Muscovite and biotite 40Ar/39Ar ages fall mainly in the interval 1400-1340 Ma, consistent with cooling through the closure temperature interval of micas (~400??-300??C) after about 1400 Ma. In contrast, hornblende apparent ages (T(c)~500??-550??C) between 1600 and 1390 Ma reflect variable retention of radiogenic argon. Forward modeling of argon diffusion shows that the distribution of hornblende and mica ages is consistent with the partial resetting of argon systematics ca. 1400 Ma by a thermal pulse reaching maximum temperatures around 550??C and decaying within <20 m.yr. These temperatures match the conditions inferred from the overprinting assemblage; thus, muscovite and biotite ages are interpreted to date the cooling phase of this metamorphic event. This late metamorphism is broadly coeval with the intrusion of ca. 1400-Ma granitic plutons in the study area and throughout the southwestern United States. However, thermal effects are observed far from pluton margins, suggesting pervasive, regional crustal heating rather than restricted contact metamorphism. Our results suggest that ca. 1400-Ma metamorphism and plutonism are manifestations of a regional thermal episode that both partially melted the lower crust and pervasively metamorphosed middle crustal rocks.

  2. Refining lunar impact chronology through high spatial resolution (40)Ar/(39)Ar dating of impact melts.

    PubMed

    Mercer, Cameron M; Young, Kelsey E; Weirich, John R; Hodges, Kip V; Jolliff, Bradley L; Wartho, Jo-Anne; van Soest, Matthijs C

    2015-02-01

    Quantitative constraints on the ages of melt-forming impact events on the Moon are based primarily on isotope geochronology of returned samples. However, interpreting the results of such studies can often be difficult because the provenance region of any sample returned from the lunar surface may have experienced multiple impact events over the course of billions of years of bombardment. We illustrate this problem with new laser microprobe (40)Ar/(39)Ar data for two Apollo 17 impact melt breccias. Whereas one sample yields a straightforward result, indicating a single melt-forming event at ca. 3.83 Ga, data from the other sample document multiple impact melt-forming events between ca. 3.81 Ga and at least as young as ca. 3.27 Ga. Notably, published zircon U/Pb data indicate the existence of even older melt products in the same sample. The revelation of multiple impact events through (40)Ar/(39)Ar geochronology is likely not to have been possible using standard incremental heating methods alone, demonstrating the complementarity of the laser microprobe technique. Evidence for 3.83 Ga to 3.81 Ga melt components in these samples reinforces emerging interpretations that Apollo 17 impact breccia samples include a significant component of ejecta from the Imbrium basin impact. Collectively, our results underscore the need to quantitatively resolve the ages of different melt generations from multiple samples to improve our current understanding of the lunar impact record, and to establish the absolute ages of important impact structures encountered during future exploration missions in the inner Solar System.

  3. Results of 40Ar/39Ar dating of phlogopites from kelyphitic rims around garnet grains (Udachnaya-Vostochnaya kimberlite pipe)

    NASA Astrophysics Data System (ADS)

    Yudin, D. S.; Tomilenko, A. A.; Alifirova, T. A.; Travin, A. V.; Murzintsev, N. G.; Pokhilenko, N. P.

    2016-07-01

    40Ar/39Ar dating of phlogopite from kelyphitic rims around garnet grains from the Udachnaya-Vostochnaya kimberlite pipe in the Sakha (Yakutia) Republic (Russia) revealed that when this mineral has contact with a kimberlite melt its age corresponds (within error limits) to that of the formation of the kimberlite pipe, thus indicating that the method may be used for dating kimberlites and related rocks. In mantle xenoliths, kelyphitic phlogopites rimming garnet grains partially lose radiogenic Ar, which results in a complex age spectrum. Rejuvenation of the K/Ar system in them is determined by the thermal impact of the kimberlite melt on captured rocks.

  4. High-resolution 40Ar 39Ar chronology of Oligocene volcanic rocks, San Juan Mountains, Colorado

    USGS Publications Warehouse

    Lanphere, M.A.

    1988-01-01

    The central San Juan caldera complex consists of seven calderas from which eight major ash-flow tuffs were erupted during a period of intense volcanic activity that lasted for approximately 2 m.y. about 26-28 Ma. The analytical precision of conventional K-Ar dating in this time interval is not sufficient to unambiguously resolve this complex history. However, 40Ar 39Ar incremental-heating experiments provide data for a high-resolution chronology that is consistent with stratigraphie relations. Weighted-mean age-spectrum plateau ages of biotite and sanidine are the most precise with standard deviations ranging from 0.08 to 0.21 m.y. The pooled estimate of standard deviation for the plateau ages of 12 minerals is about 0.5 percent or about 125,000 to 135,000 years. Age measurements on coexisting minerals from one tuff and on two samples of each of two other tuffs indicate that a precision in the age of a tuff of better than 100,000 years can be achieved at 27 Ma. New data indicate that the San Luis caldera is the youngest caldera in the central complex, not the Creede caldera as previously thought. ?? 1988.

  5. 40Ar/39Ar Ages for the Sentinel-Arlington Volcanic Field, Southwestern Arizona

    NASA Astrophysics Data System (ADS)

    Cave, S. R.; Greeley, R.; Champion, D. E.; Turrin, B. D.

    2007-12-01

    .16-51.48. Geochronology using 40Ar/39Ar method revealed an age of 1.94 +/- 0.85 Ma for Painted Rock Low Shield (New Mexico Geochronology Research Laboratory), 1.64 +/- 0.14 Ma for Theba Low Shield (Rutgers University) and 1.24 +/- 0.040 Ma for Wild Horse Low Shield (Rutgers University). Some ages were precise enough to correspond to the Matuyama reversed polarity epoch, with SAVF initiation possibly within the Olduvai normal polarity event. These dates represent an overall improvement in precision and accuracy over previous dates (values corresponding to 6.20 Ma to 1.28 Ma) collected in the late 1970s and early 1980s using K-Ar technique. The 40Ar/39Ar ages correspond to expected magnetic polarities and stratigraphic sequences.

  6. Methylenetetrahydrofolate Reductase C677T: Hypoplastic Left Heart and Thrombosis.

    PubMed

    Spronk, Kimberly J; Olivero, Anthony D; Haw, Marcus P; Vettukattil, Joseph J

    2015-10-01

    The incidence of congenital heart defects is higher in infants with mutation of methylenetetrahydrofolate reductase (MTHFR) gene. The MTHFR C677T gene decreases the bioavailability of folate and increases plasma homocysteine, a risk factor for thrombosis. There have been no reported cases in the literature on the clinical implications of this procoagulable state in the setting of cyanotic heart disease, which itself has prothrombotic predisposition. Two patients with hypoplastic left heart syndrome developed postoperative thrombotic complications, both were homozygous for MTHFR C677T. We present these cases and highlight the implications of MTHFR mutation in the management of complex congenital heart disease. PMID:26467879

  7. Terpenoids from Diplophyllum taxifolium with quinone reductase-inducing activity.

    PubMed

    Wang, Xiao; Zhang, Jiao-Zhen; Zhou, Jin-Chuan; Shen, Tao; Lou, Hong-Xiang

    2016-03-01

    Two new ent-prenylaromadendrane-type diterpenoids, diplotaxifols A (1) and B (2), a new ent-eudesmol, ent-eudesma-4(15),11(13)-dien-6α,12-diol (3), eight new eudesmanolides enantiomers (4-11) of the corresponding compounds from higher plants along with four known ent-eudesmanolides (12-15) were isolated from the 95% EtOH extract of Chinese liverwort Diplophyllum taxifolium. Their structures were elucidated on the basis of MS, NMR and IR spectral data, and confirmed by single-crystal X-ray diffraction analysis. The quinone reductase-inducing activity of the compounds was evaluated. PMID:26656409

  8. Differential Light Induction of Nitrate Reductases in Greening and Photobleached Soybean Seedlings 1

    PubMed Central

    Kakefuda, Genichi; Duke, Stanley H.; Duke, Stephen O.

    1983-01-01

    Soybean (Glycine max [L.] Merr.) seeds were imbibed and germinated with or without NO3−, tungstate, and norflurazon (San 9789). Norflurazon is a herbicide which causes photobleaching of chlorophyll by inhibiting carotenoid synthesis and which impairs normal chloroplast development. After 3 days in the dark, seedlings were placed in white light to induce extractable nitrate reductase activity. The induction of maximal nitrate reductase activity in greening cotyledons did not require NO3− and was not inhibited by tungstate. Induction of nitrate reductase activity in norflurazon-treated cotyledons had an absolute requirement for NO3− and was completely inhibited by tungstate. Nitrate was not detected in seeds or seedlings which had not been treated with NO3−. The optimum pH for cotyledon nitrate reductase activity from norflurazon-treated seedlings was at pH 7.5, and near that for root nitrate reductase activity, whereas the optimum pH for nitrate reductase activity from greening cotyledons was pH 6.5. Induction of root nitrate reductase activity was also inhibited by tungstate and was dependent on the presence of NO3−, further indicating that the isoform of nitrate reductase induced in norflurazon-treated cotyledons is the same or similar to that found in roots. Nitrate reductases with and without a NO3− requirement for light induction appear to be present in developing leaves. In vivo kinetics (light induction and dark decay rates) and in vitro kinetics (Arrhenius energies of activation and NADH:NADPH specificities) of nitrate reductases with and without a NO3− requirement for induction were quite different. Km values for NO3− were identical for both nitrate reductases. PMID:16663185

  9. Synthesis and activity of novel 16-dehydropregnenolone acetate derivatives as inhibitors of type 1 5α-reductase and on cancer cell line SK-LU-1.

    PubMed

    Silva-Ortiz, Aylin Viviana; Bratoeff, Eugene; Ramírez-Apan, Teresa; Heuze, Yvonne; Sánchez, Araceli; Soriano, Juan; Cabeza, Marisa

    2015-12-15

    Testosterone (T) plays a crucial role in prostate growth. In androgen-dependent tissues T is reduced to dihydrotestosterone (DHT) because of the presence of the 5α-reductase enzyme. This androgen is more active than T, since it has a higher affinity for the androgen receptor (AR). When this mechanism is altered, androgen-dependent diseases, including prostate cancer, could result. The aim of this study was to synthesize several 16-dehydropregnenolone acetate derivatives containing a triazole ring at C-21 and a linear or alicyclic ester moiety at C-3 of the steroidal skeleton. These steroids were designed as potential inhibitors of the activity of both types (1 and 2) of 5α-reductase. The cytotoxic activity of these compounds was also evaluated on a panel of PC-3, MCF7, and SK-LU-1 human cancer cell lines. The results from this study showed that with the exception of steroids 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-propionate and 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-pentanoate, the compounds exhibit a lower inhibitory activity for both isoenzymes of 5α-reductase than finasteride. Furthermore the 3β-hydroxy-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-20-one and 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-acetate derivatives display 80% cytotoxic activity on the SK-LU-1 cell line. These results also indicated that the triazole derivatives, which have a hydroxyl or acetoxy group at C-3, could have an anticancer effect, whereas the derivatives with a alicyclic ester group at C-3 do not show biological activity.

  10. Synthesis and activity of novel 16-dehydropregnenolone acetate derivatives as inhibitors of type 1 5α-reductase and on cancer cell line SK-LU-1.

    PubMed

    Silva-Ortiz, Aylin Viviana; Bratoeff, Eugene; Ramírez-Apan, Teresa; Heuze, Yvonne; Sánchez, Araceli; Soriano, Juan; Cabeza, Marisa

    2015-12-15

    Testosterone (T) plays a crucial role in prostate growth. In androgen-dependent tissues T is reduced to dihydrotestosterone (DHT) because of the presence of the 5α-reductase enzyme. This androgen is more active than T, since it has a higher affinity for the androgen receptor (AR). When this mechanism is altered, androgen-dependent diseases, including prostate cancer, could result. The aim of this study was to synthesize several 16-dehydropregnenolone acetate derivatives containing a triazole ring at C-21 and a linear or alicyclic ester moiety at C-3 of the steroidal skeleton. These steroids were designed as potential inhibitors of the activity of both types (1 and 2) of 5α-reductase. The cytotoxic activity of these compounds was also evaluated on a panel of PC-3, MCF7, and SK-LU-1 human cancer cell lines. The results from this study showed that with the exception of steroids 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-propionate and 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-pentanoate, the compounds exhibit a lower inhibitory activity for both isoenzymes of 5α-reductase than finasteride. Furthermore the 3β-hydroxy-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-20-one and 20-oxo-21-(1H-1,2,4-triazole-1-yl)pregna-5,16-dien-3β-yl-acetate derivatives display 80% cytotoxic activity on the SK-LU-1 cell line. These results also indicated that the triazole derivatives, which have a hydroxyl or acetoxy group at C-3, could have an anticancer effect, whereas the derivatives with a alicyclic ester group at C-3 do not show biological activity. PMID:26631442

  11. Di-(2-ethylhexyl) phthalate inhibits testosterone level through disturbed hypothalamic-pituitary-testis axis and ERK-mediated 5α-Reductase 2.

    PubMed

    Ha, Mei; Guan, Xie; Wei, Li; Li, Peng; Yang, Min; Liu, Changjiang

    2016-09-01

    Di-(2-ethylhexyl) phthalate (DEHP) has reproductive toxicity and can affect male reproductive development. In order to clarify adverse effects of DEHP on testicular physiology and testosterone production, Sprague-Dawley (SD) rats were dosed daily with DEHP by gavage for 30days; TM3 cells (mouse Leydig cell line) were treated with DEHP for 24h after pretreatment with vitamin C or U0126. Results indicated that the hypothalamic-pituitary-testis (HPT) axis was disturbed and serum testosterone, LH and FSH levels were decreased following DEHP exposure. Histomorphological changes of rat testes were also observed, such as deformed seminiferous tubules, aggregated chromatin, multiple vacuoles, swollen mitochondria, apoptotic germ cells and Sertoli cells, as well as increased Leydig cell numbers. Moreover, DEHP caused oxidative stress in vivo and in vitro and then induced the ERK pathway, which was required to mediate 5α-Reductase 2 and scavenger receptor class B-1 (SRB1) levels. However, levels of steroidogenic acute regulatory protein (StAR), 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-hydroxysteroid dehydrogenase (17β-HSD), P450 17α-hydroxylase/17.20 lyase (P450c17), and P450 side-chain cleavage enzyme (P450scc) were not significantly altered after DEHP exposure. Taken together, DEHP-disturbed HPT axis and induced 5α-Reductase 2 contribute to the reduction of serum testosterone level. The activated ERK pathway is required to modulate expressions of 5α-Reductase 2 and SRB1. PMID:27155079

  12. Androgen Receptor (AR) Physiological Roles in Male and Female Reproductive Systems: Lessons Learned from AR-Knockout Mice Lacking AR in Selective Cells1

    PubMed Central

    Chang, Chawnshang; Lee, Soo Ok; Wang, Ruey-Sheng; Yeh, Shuyuan; Chang, Ta-Min

    2013-01-01

    ABSTRACT Androgens/androgen receptor (AR) signaling is involved primarily in the development of male-specific phenotypes during embryogenesis, spermatogenesis, sexual behavior, and fertility during adult life. However, this signaling has also been shown to play an important role in development of female reproductive organs and their functions, such as ovarian folliculogenesis, embryonic implantation, and uterine and breast development. The establishment of the testicular feminization (Tfm) mouse model exploiting the X-linked Tfm mutation in mice has been a good in vivo tool for studying the human complete androgen insensitivity syndrome, but this mouse may not be the perfect in vivo model. Mouse models with various cell-specific AR knockout (ARKO) might allow us to study AR roles in individual types of cells in these male and female reproductive systems, although discrepancies are found in results between labs, probably due to using various Cre mice and/or knocking out AR in different AR domains. Nevertheless, no doubt exists that the continuous development of these ARKO mouse models and careful studies will provide information useful for understanding AR roles in reproductive systems of humans and may help us to develop more effective and more specific therapeutic approaches for reproductive system-related diseases. PMID:23782840

  13. Eruption age of an approximately 100,000-year-old basalt from Ar-40/Ar-39 analysis of partially degassed xenoliths

    NASA Technical Reports Server (NTRS)

    Gillespie, A. R.; Huneke, J. C.; Wasserburg, G. J.

    1984-01-01

    Eleven samples from five partially degassed Mesozoic granitic xenoliths and two late Pleistocene host basalt samples from Sawmill Canyon in Sierra Nevada, California, have been analyzed using stepwise Ar-40/Ar-39 dating techniques. The petrography and analytic techniques are described and the results are presented. Isotopic compositions of Ar released from the xenoliths in several extraction steps at temperatures below about 900 C were colinear in Ar-36/Ar-40 versus Ar-39/Ar-40 diagrams and defined isochrons, giving a mean age of degassing of 119,000 + or - 7000 yr. Ar-40 extracted at higher temperatures included ancient radiogenic Ar-40 that never diffused from the xenoliths during immersion in the magma. This Ar-40 caused an increase in the apparent age for the high-temperature extractions. The high precision of the eruption age determined by this method is comparable to that obtained elsewhere by conventional K/Ar dating of sanidine.

  14. Effect of surface derived hydrocarbon impurities on Ar plasma properties

    SciTech Connect

    Fox-Lyon, Nick; Oehrlein, Gottlieb S.; Godyak, Valery

    2014-05-15

    The authors report on Langmuir probe measurements that show that hydrocarbon surfaces in contact with Ar plasma cause changes of electron energy distribution functions due to the flux of hydrogen and carbon atoms released by the surfaces. The authors compare the impact on plasma properties of hydrocarbon species gasified from an etching hydrocarbon surface with injection of gaseous hydrocarbons into Ar plasma. They find that both kinds of hydrocarbon injections decrease electron density and slightly increase electron temperatures of low pressure Ar plasma. For low percentages of impurities (∼1% impurity in Ar plasma explored here), surface-derived hydrocarbon species and gas phase injected hydrocarbon molecules cause similar changes of plasma properties for the same number of hydrocarbon molecules injected into Ar with a decrease in electron density of ∼4%.

  15. A SAM-dependent methyltransferase cotranscribed with arsenate reductase alters resistance to peptidyl transferase center-binding antibiotics in Azospirillum brasilense Sp7.

    PubMed

    Singh, Sudhir; Singh, Chhaya; Tripathi, Anil Kumar

    2014-05-01

    The genome of Azospirillum brasilense harbors a gene encoding S-adenosylmethionine-dependent methyltransferase, which is located downstream of an arsenate reductase gene. Both genes are cotranscribed and translationally coupled. When they were cloned and expressed individually in an arsenate-sensitive strain of Escherichia coli, arsenate reductase conferred tolerance to arsenate; however, methyltransferase failed to do so. Sequence analysis revealed that methyltransferase was more closely related to a PrmB-type N5-glutamine methyltransferase than to the arsenate detoxifying methyltransferase ArsM. Insertional inactivation of prmB gene in A. brasilense resulted in an increased sensitivity to chloramphenicol and resistance to tiamulin and clindamycin, which are known to bind at the peptidyl transferase center (PTC) in the ribosome. These observations suggested that the inability of prmB:km mutant to methylate L3 protein might alter hydrophobicity in the antibiotic-binding pocket of the PTC, which might affect the binding of chloramphenicol, clindamycin, and tiamulin differentially. This is the first report showing the role of PrmB-type N5-glutamine methyltransferases in conferring resistance to tiamulin and clindamycin in any bacterium.

  16. Search for cosmogenic Ar-42 in meteorites

    NASA Astrophysics Data System (ADS)

    Cini Castagnoli, G.; Cane, D.; Taricco, C.; Bhandari, N.

    2003-04-01

    We have evidence for decreasing galactic cosmic ray (GCR) flux in the past 3 centuries by a factor about two [1]. The measurements of the activity of cosmogenic 44Ti (T1/2 = 59.2 y) produced by GCR in stony meteorites that fell during the last two centuries are in good agreement with the calculated values and validate the decreasing trend of GCR flux. The measurements were obtained by an hyperpure (372 cm3) Ge-NaI(Tl) spectrometer operating in the Monte dei Cappuccini laboratory in Torino[2]. To further improve upon statistical precision of 44Ti data and also to be able to measure the 42Ar (T1/2 = 33 y) which is produced in even smaller amounts by GCR in meteorites, we have set up in the same laboratory a larger (581 cm3) hyperpure Ge detector operating in coincidence with a 100 kg NaI(Tl) crystal assembly. This should enable us to confirm the above results about GCR variations. We wish to acknowledge our deep gratitude to professor Bonino, deceased on September 29, 2002, to whom the assemblage of the new equipment is due. [1] G. Bonino, G. Cini Castagnoli, D. Cane, C. Taricco and N. Bhandari,Proc. XXVII Intern. Cosmic Ray Conf. (Hamburg, 2001) 3769-3772. [2] Bonino G., Cini Castagnoli G., Cane D., Taricco C., Bhandari N, textit {34th COSPAR Sci. Ass. Houston 2002 (Adv. Space Res.)}, in press

  17. A compilation of K-Ar-ages for southern California

    USGS Publications Warehouse

    Miller, Fred K.; Morton, Douglas M.; Morton, Janet L.; Miller, David M.

    2014-01-01

    The purpose of this report is to make available a large body of conventional K-Ar ages for granitic, volcanic, and metamorphic rocks collected in southern California. Although one interpretive map is included, the report consists primarily of a systematic listing, without discussion or interpretation, of published and unpublished ages that may be of value in future regional and other geologic studies. From 1973 to 1979, 468 rock samples from southern California were collected for conventional K-Ar dating under a regional geologic mapping project of Southern California (predecessor of the Southern California Areal Mapping Project). Most samples were collected and dated between 1974 and 1977. For 61 samples (13 percent of those collected), either they were discarded for varying reasons, or the original collection data were lost. For the remaining samples, 518 conventional K-Ar ages are reported here; coexisting mineral pairs were dated from many samples. Of these K-Ar ages, 225 are previously unpublished, and identified as such in table 1. All K-Ar ages are by conventional K-Ar analysis; no 40Ar/39Ar dating was done. Subsequent to the rock samples collected in the 1970s and reported here, 33 samples were collected and 38 conventional K-Ar ages determined under projects directed at (1) characterization of the Mesozoic and Cenozoic igneous rocks in and on both sides of the Transverse Ranges and (2) clarifying the Mesozoic and Cenozoic tectonics of the eastern Mojave Desert. Although previously published (Beckerman et al., 1982), another eight samples and 11 conventional K-Ar ages are included here, because they augment those completed under the previous two projects.

  18. A Novel NADPH-dependent flavoprotein reductase from Bacillus megaterium acts as an efficient cytochrome P450 reductase.

    PubMed

    Milhim, Mohammed; Gerber, Adrian; Neunzig, Jens; Hannemann, Frank; Bernhardt, Rita

    2016-08-10

    Cytochromes P450 (P450s) require electron transfer partners to catalyze substrate conversions. With regard to biotechnological approaches, the elucidation of novel electron transfer proteins is of special interest, as they can influence the enzymatic activity and specificity of the P450s. In the current work we present the identification and characterization of a novel soluble NADPH-dependent diflavin reductase from Bacillus megaterium with activity towards a bacterial (CYP106A1) and a microsomal (CYP21A2) P450 and, therefore, we referred to it as B. megaterium cytochrome P450 reductase (BmCPR). Sequence analysis of the protein revealed besides the conserved FMN-, FAD- and NADPH-binding motifs, the presence of negatively charged cluster, which is thought to represent the interaction domain with P450s and/or cytochrome c. BmCPR was expressed and purified to homogeneity in Escherichia coli. The purified BmCPR exhibited a characteristic diflavin reductase spectrum, and showed a cytochrome c reducing activity. Furthermore, in an in vitro reconstituted system, the BmCPR was able to support the hydroxylation of testosterone and progesterone with CYP106A1 and CYP21A2, respectively. Moreover, in view of the biotechnological application, the BmCPR is very promising, as it could be successfully utilized to establish CYP106A1- and CYP21A2-based whole-cell biotransformation systems, which yielded 0.3g/L hydroxy-testosterone products within 8h and 0.16g/L 21-hydroxyprogesterone within 6h, respectively. In conclusion, the BmCPR reported herein owns a great potential for further applications and studies and should be taken into consideration for bacterial and/or microsomal CYP-dependent bioconversions.

  19. 40Ar-39Ar Results of Lunar Meteorites Dhofar 025, 280, 309, 730, 733, 1436, 1442, SAU 449, NWA 6888

    NASA Astrophysics Data System (ADS)

    Korochantseva, E. V.; Buikin, A. I.; Hopp, J.; Korochantsev, A. V.; Trieloff, M.

    2016-08-01

    40Ar-39Ar ages vary between 3.0 and 4.2 Ga common for lunar rocks, but only Dho 025, Dho 309 and Dho 1436 are consistent with the LHB. Dho 280, 733, 1442, and NWA 6888 were affected by young impact events ≤1 Ga ago, partially also reset of CRE ages.

  20. Paleomagnetic and 40Ar/39Ar geochronologic data from late Proterozoic mafic dikes and sills, Montana and Wyoming

    USGS Publications Warehouse

    Harlan, Stephen S.; Geissman, John William; Snee, Lawrence W.

    1997-01-01

    Paleomagnetic and 40Ar/39Ar results from mafic dikes and sills in northwestern Wyoming and western Montana yield similar virtual geomagnetic poles and isotopic dates. In combination with paleomagnetic and geochronologic data from elsewhere in the western Cordillera, these data provide evidence for a regional mafic magnetic event at 780 to 770 Ma that affected a large area of western North America.

  1. On Full Disclosure and Transparent Data Flow from 40Ar/39Ar Geochronology Measurements to Data Reduction to Online Repositories

    NASA Astrophysics Data System (ADS)

    Koppers, A. A. P.

    2015-12-01

    Arguably 40Ar/39Ar geochronology is one of the most versatile techniques available to Earth scientists today for the dating of rocks and minerals and determining the rates of geological processes on Earth and in our solar system. Over the last four decades large quantities of high (and lower) quality 40Ar/39Ar data have been produced using many different generations of mass spectrometry instrumentation. This wealth of data is only as useful as its description and availability of metadata allows. Many online data sets or compilations available in the science literature only carry the resulting product, an age and a related uncertainty in millions of years, for example. These data points are far from desirable as these don't allow recalculation against modern-day age standards, decay constants and other parameters essential in 40Ar/39Ar geochronology. Over time these data will become less useful to the research community and eventually these will be put by the wayside. In this presentation I will emphasize the need for full disclosure of all data and metadata involved in 40Ar/39Ar geochronology. I will give examples of how a complex data flow can be kept transparent from sample preparation to measurement to data reduction and eventually the uploading into online data repositories. Without the full disclosure of our data and a transparent data flow, it is evident that we cannot live up to one of the governing doctrines in the sciences, namely reproducibility of our scientific experiments and findings.

  2. Differential distribution of cAMP receptors cAR2 and cAR3 during Dictyostelium development.

    PubMed

    Yu, Y; Saxe, C L

    1996-01-10

    Signal transduction via a family of cAMP receptor subtypes (cARs) is critical for proper development in the cellular slime mold Dictyostelium. Genes encoding four related subtypes have been cloned and their expression, based on RNA accumulation, has been previously reported. Here we report the differential spatial and temporal distribution of cAR2 and cAR3 proteins, based on indirect double immunofluorescence. Cells were transformed with a carB::lacZ construct, and an antibody against beta-galactosidase was used to visualize cAR2 expression. Simultaneously, a cAR3-specific antibody was used to identify cAR3-expressing cells. Results indicate that by the time of tip formation (12-14 hr) both receptors are expressed and distribute in a virtually nonoverlapping pattern, with cAR2 being expressed on anterior, prestalk cells and cAR3 present in the rest of the organism. Differential distribution of these two receptor subtypes may result in distinct cAMP signaling mechanisms in the two major regions of the organism. PMID:8575636

  3. Sequence and properties of pentaerythritol tetranitrate reductase from Enterobacter cloacae PB2.

    PubMed Central

    French, C E; Nicklin, S; Bruce, N C

    1996-01-01

    Pentaerythritol tetranitrate reductase, which reductively liberates nitrite from nitrate esters, is related to old yellow enzyme. Pentaerythritol tetranitrate reductase follows a ping-pong mechanism with competitive substrate inhibition by NADPH, is strongly inhibited by steroids, and is capable of reducing the unsaturated bond of 2-cyclohexen-1-one. PMID:8932320

  4. Determination of the specific activities of methionine sulfoxide reductase A and B by capillary electrophoresis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A capillary electrophoresis (CE) method for the determination of methionine sulfoxide reductase A and methionine sulfoxide reductase B activities in mouse liver is described. The method is based on detection of the 4-(dimethylamino)azobenzene-4’-sulfonyl derivative of L-methionine (dabsyl Met), the ...

  5. Regulation of a ribonucleoside reductase during the early generative phase in Acetabularia.

    PubMed

    de Groot, E J; Schweiger, H G

    1985-02-01

    The activity of a ribonucleoside reductase was estimated during the life cycle of Acetabularia. During the early generative phase the enzyme activity was dramatically increased. Regulation of the ribonucleoside reductase was observed even in the absence of the nucleus. The increase in activity was inhibited by chloramphenicol but not by cycloheximide. These results indicate that the enzyme is translated on 70 S ribosomes.

  6. Nitrate transport is independent of NADH and NAD(P)H nitrate reductases in barley seedlings

    NASA Technical Reports Server (NTRS)

    Warner, R. L.; Huffaker, R. C.

    1989-01-01

    Barley (Hordeum vulgare L.) has NADH-specific and NAD(P)H-bispecific nitrate reductase isozymes. Four isogenic lines with different nitrate reductase isozyme combinations were used to determine the role of NADH and NAD(P)H nitrate reductases on nitrate transport and assimilation in barley seedlings. Both nitrate reductase isozymes were induced by nitrate and were required for maximum nitrate assimilation in barley seedlings. Genotypes lacking the NADH isozyme (Az12) or the NAD(P)H isozyme (Az70) assimilated 65 or 85%, respectively, as much nitrate as the wild type. Nitrate assimilation by genotype (Az12;Az70) which is deficient in both nitrate reductases, was only 13% of the wild type indicating that the NADH and NAD(P)H nitrate reductase isozymes are responsible for most of the nitrate reduction in barley seedlings. For all genotypes, nitrate assimilation rates in the dark were about 55% of the rates in light. Hypotheses that nitrate reductase has direct or indirect roles in nitrate uptake were not supported by this study. Induction of nitrate transporters and the kinetics of net nitrate uptake were the same for all four genotypes indicating that neither nitrate reductase isozyme has a direct role in nitrate uptake in barley seedlings.

  7. SORGOdb: Superoxide Reductase Gene Ontology curated DataBase

    PubMed Central

    2011-01-01

    Background Superoxide reductases (SOR) catalyse the reduction of superoxide anions to hydrogen peroxide and are involved in the oxidative stress defences of anaerobic and facultative anaerobic organisms. Genes encoding SOR were discovered recently and suffer from annotation problems. These genes, named sor, are short and the transfer of annotations from previously characterized neelaredoxin, desulfoferrodoxin, superoxide reductase and rubredoxin oxidase has been heterogeneous. Consequently, many sor remain anonymous or mis-annotated. Description SORGOdb is an exhaustive database of SOR that proposes a new classification based on domain architecture. SORGOdb supplies a simple user-friendly web-based database for retrieving and exploring relevant information about the proposed SOR families. The database can be queried using an organism name, a locus tag or phylogenetic criteria, and also offers sequence similarity searches using BlastP. Genes encoding SOR have been re-annotated in all available genome sequences (prokaryotic and eukaryotic (complete and in draft) genomes, updated in May 2010). Conclusions SORGOdb contains 325 non-redundant and curated SOR, from 274 organisms. It proposes a new classification of SOR into seven different classes and allows biologists to explore and analyze sor in order to establish correlations between the class of SOR and organism phenotypes. SORGOdb is freely available at http://sorgo.genouest.org/index.php. PMID:21575179

  8. Retrospective approach to methylenetetrahydrofolate reductase mutations in children.

    PubMed

    Özer, Işıl; Özçetin, Mustafa; Karaer, Hatice; Kurt, Semiha G; Şahin, Şemsettin

    2011-07-01

    Methylenetetrahydrofolate reductase reduces methyltetrahydrofolate, a cosubstrate in the remethylation of homocysteine, from methylenetetrahydrofolate. Congenital defects, hematologic tumors, and intrauterine growth retardation can occur during childhood. This study evaluated clinical and laboratory treatment approaches in children diagnosed with methylenetetrahydrofolate reductase mutations. Our group included 23 boys and 14 girls, aged 103.4 ± 70.8 months S.D. Clinical findings of patients and homocysteine, vitamin B12, folate, hemogram, electroencephalography, cranial magnetic resonance imaging, and echocardiography data were evaluated in terms of treatment approach. Our patients' findings included vitamin B12 at 400.4 ± 224.6 pg/mL S.D. (normal range, 300-700 pg/mL), folate at 10.1 ± 4.5 ng/mL S.D. (normal range, 1.8-9 ng/mL), and homocysteine at 8.4 ± 4.7 μmol/L S.D. (normal range, 5.5-17 μmol/L). Eighty-eight percent of patients demonstrated clinical findings. In comparisons involving categorical variables between groups, χ(2) tests were used. No relationship was evident between mutation type, laboratory data, and clinical severity. All mothers who had MTHFR mutations and had babies with sacral dimples had taken folate supplements during pregnancy. To avoid the risk of neural tube defects, pregnant women with a MTHFR mutation may require higher than normally recommended doses of folic acid supplementation for optimum health.