Science.gov

Sample records for 12cgamma 3alpha ehnergeticheskoe

  1. Simultaneous quantification of GABAergic 3alpha,5alpha/3alpha,5beta neuroactive steroids in human and rat serum.

    PubMed

    Porcu, Patrizia; O'Buckley, Todd K; Alward, Sarah E; Marx, Christine E; Shampine, Lawrence J; Girdler, Susan S; Morrow, A Leslie

    2009-01-01

    The 3alpha,5alpha- and 3alpha,5beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone enhance GABAergic neurotransmission and produce inhibitory neurobehavioral and anti-inflammatory effects. Despite substantial information on the progesterone derivative (3alpha,5alpha)-3-hydroxypregnan-20-one (3alpha,5alpha-THP, allopregnanolone), the physiological significance of the other endogenous GABAergic neuroactive steroids has remained elusive. Here, we describe the validation of a method using gas chromatography-mass spectrometry to simultaneously identify serum levels of the eight 3alpha,5alpha- and 3alpha,5beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone. The method shows specificity, sensitivity and enhanced throughput compared to other methods already available for neuroactive steroid quantification. Administration of pregnenolone to rats and progesterone to women produced selective effects on the 3alpha,5alpha- and 3alpha,5beta-reduced neuroactive steroids, indicating differential regulation of their biosynthetic pathways. Pregnenolone administration increased serum levels of 3alpha,5alpha-THP (+1488%, p<0.001), (3alpha,5alpha)-3,21-dihydroxypregnan-20-one (3alpha,5alpha-THDOC, +205%, p<0.01), (3alpha,5alpha)-3-hydroxyandrostan-17-one (3alpha,5alpha-A, +216%, p<0.001), (3alpha,5alpha,17beta)-androstane-3,17-diol (3alpha,5alpha-A-diol, +190%, p<0.01). (3alpha,5beta)-3-hydroxypregnan-20-one (3alpha,5beta-THP) and (3alpha,5beta)-3-hydroxyandrostan-17-one (3alpha,5beta-A) were not altered, while (3alpha,5beta)-3,21-dihydroxypregnan-20-one (3alpha,5beta-THDOC) and (3alpha,5beta,17beta)-androstane-3,17-diol (3alpha,5beta-A-diol) were increased from undetectable levels to 271+/-100 and 2.4+/-0.9 pg+/-SEM, respectively (5/8 rats). Progesterone administration increased serum levels of 3alpha,5alpha-THP (+1806%, p<0.0001), 3alpha,5beta-THP (+575%, p<0.001), 3alpha,5alpha

  2. Activating STAT3 Alpha for Promoting Healing of Neurons

    NASA Technical Reports Server (NTRS)

    Conway, Greg

    2008-01-01

    A method of promoting healing of injured or diseased neurons involves pharmacological activation of the STAT3 alpha protein. Usually, injured or diseased neurons heal incompletely or not at all for two reasons: (1) they are susceptible to apoptosis (cell death); and (2) they fail to engage in axogenesis that is, they fail to re-extend their axons to their original targets (e.g., muscles or other neurons) because of insufficiency of compounds, denoted neurotrophic factors, needed to stimulate such extension. The present method (see figure) of treatment takes advantage of prior research findings to the effect that the STAT3 alpha protein has anti-apoptotic and pro-axogenic properties.

  3. Brassinolide activities of 2alpha,3alpha-diols versus 3alpha,4alpha-diols in the bean second internode bioassay: explanation by molecular modeling methods.

    PubMed

    Sísa, Miroslav; Vilaplana-Polo, Marc; Ballesteros, Carme Brosa; Kohout, Ladislav

    2007-10-01

    In general, the structural requirements postulated for a high brassinolide activity are: 2alpha,3alpha-diol, 6-ketone or better 7-oxalactone in B-ring, A/B trans fused ring junction, a cis C-22,C-23-diol preferentially with RR configurations, and a C-24 methyl or ethyl substituent [Takatsuto S, Yazawa N, Ikekawa N, Takematsu T, Takeuchi Y, Koguchi M. Structure-activity relationship of brassinosteroids. Phytochemistry 1983;22:2437-41; Thompson MJ, Meudt WJ, Mandava NB, Dutky SR, Lusby WR, Spaulding DW. Synthesis of brassinosteroids and relationship of structure to plant growth-promoting effects. Steroids 1982;39:89-105]. We found that the 3alpha,4alpha-diols 4, 6 and 8 are more active than the 2alpha,3alpha-diols 3, 5 and 7 [Sísa M, Budesínský M, Kohout L. Synthesis of 7a-homo and 7a,7b-dihomo-5alpha-cholestane analogues of brassinolide. Collect Czech Chem Commun 2003;68:2171-89]. This fact is in strong contrast with the structure requirements mentioned above. Our hypothesis suggests that the lower activity of 2alpha,3alpha-diols and/or the higher activity of 3alpha,4alpha-diols could be explained by twisting and distortion of the molecule due to the seven- or eight-membered B-ring and also by the position of a carbonyl group relative to the A-ring diol. 3D-SAR computer methodologies as alignments and overlaps of GRID maps and 3D-QSAR analysis GRID-GOLPE (CoMFA-like) were used as an effort to explain the higher bioactivity of 3alpha,4alpha-diols 4, 6 and 8 in comparison with the 2alpha,3alpha-diols 3, 5 and 7 of B-ring enlarged brassinosteroids.

  4. 3 alpha-hydroxysteroid dehydrogenase: three dimensional structure and gene regulation.

    PubMed

    Penning, T M

    1996-09-01

    Mammalian 3 alpha-hydroxysteroid dehydrogenases (3 alpha-HSDs) regulate steroid hormone levels. cDNA cloning indicates that the rat and human liver isoforms display high sequence identity and that they belong to the aldo-keto reductase (AKR) superfamily. Of these the most extensively characterized is rat liver 3 alpha-HSD. The recently solved X-ray crystal structure shows that this enzyme adopts an (alpha/beta)8-barrel scaffold (Hoog et al. 1994). NAD(P)H binds in an extended anti-conformation and lies along the inner surface of the barrel. The nicotinamide ring is stabilized by interaction with Y216. The 4-pro(R)-hydrogen transferred in the reaction is in close proximity to Y55. K84, D50 and H117 which are implicated in catalysis. These residues are located at the base of a hydrophobic pocket which is presumed to be involved in binding steroid hormone. This catalytic tetrad is conserved in members of the AKR superfamily. Mutant enzymes support roles for Y55 in steroid binding and for K84 as the general acid involved in catalysis. The gene for rat 3 alpha-HSD has been cloned and is 47 kb in length and contains 9 exon-intron boundaries which are highly conserved in the human gene(s). The 5'-flanking regions of the rat and human genes contain consensus sequences for AP-1, Oct-1 and multiple copies of perfect and imperfect steroid hormone response elements (REs) (estrogen, glucocorticoid (GRE), and progesterone) which may comprise a steroid response unit (SRU) (Lin & Penning 1995). Constitutive and regulated expression of the rat 3 alpha-HSD gene has been studied by transiently transfecting reporter gene (chloramphenicol acetyltransferase, CAT) constructs into human hepatoma (HepG2) cells. With respect to the transcription start-site (+1), a proximal (-498 to -199bp) and distal (-20 to -4.0kb) enhancer, as well as a powerful silencer (-755 to -498 bp) were located in the promoter. Band-shift and supershift assays provide evidence that Oct-1 binds to the silencer

  5. Infusions of 3alpha,5alpha-THP to the VTA enhance exploratory, anti-anxiety, social, and sexual behavior and increase levels of 3alpha,5alpha-THP in midbrain, hippocampus, diencephalon, and cortex of female rats.

    PubMed

    Frye, Cheryl A; Rhodes, Madeline E

    2008-02-11

    17beta-Estradiol (E2) and progesterone (P4) influence the onset and duration of sexual behavior and are also associated with changes in behaviors that may contribute to mating, such as exploration, anxiety, and social behaviors (socio-sexual behaviors). In the midbrain ventral tegmental area (VTA), the P4 metabolite, 5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP), modulates lordosis of E2-primed rodents; 3alpha,5alpha-THP can also influence anxiety and social behaviors. To examine if 3alpha,5alpha-THP in the VTA mediates socio-sexual behaviors, we infused 3alpha,5alpha-THP to the VTA of diestrous and proestrous rats. As expected, proestrous, compared to diestrous, rats showed more exploratory (open field), anxiolytic (elevated plus maze), pro-social (partner preference, social interaction), and sexual (paced mating) behavior and had increased E2, P4, dihydroprogesterone (DHP), and 3alpha,5alpha-THP in serum, midbrain, hippocampus, diencephalon, and cortex. Infusions of 3alpha,5alpha-THP to the VTA, but not control sites, such as the substantia nigra (SN) or central grey (CG), of diestrous rats produced behavioral and endocrine effects akin to that of proestrous rats and increased DHP and 3alpha,5alpha-THP levels in midbrain, hippocampus, and diencephalon. Levels of DHP and 3alpha,5alpha-THP, but neither E2 nor P4 concentrations, in midbrain, hippocampus, diencephalon, and/or cortex were positively correlated with socio-sexual behaviors. Thus, 3alpha,5alpha-THP infusions to the VTA, but not SN or CG, can enhance socio-sexual behaviors and increase levels in midbrain, hippocampus, and diencephalon.

  6. Effects of a new 3-alpha reaction on the s-process in massive stars

    SciTech Connect

    Kikuch, Yukihiro; Ono, Masaomi; Matsuo, Yasuhide; Hashimoto, Masa-aki; Fujimoto, Shin-ichiro

    2012-11-12

    Effect of a new 3-alpha reaction rate on the s-process during the evolution of a massive star of 25 solar mass is investigated for the first time, because the s-process in massive stars have been believed to be established with only minor change. We find that the s-process with use of the new rate during the core helium burning is very inefficient compared to the case with the previous 3-alpha rate. However, the difference of the overproduction is found to be largely compensated by the subsequent carbon burning. Since the s-process in massive stars has been attributed so far to the neutron irradiation during core helium burning, our finding reveals for the first time the importance of the carbon burning for the s-process during the evolution of massive stars.

  7. Fluoxetine may influence lordosis of rats through effects on midbrain 3 alpha,5 alpha-THP concentrations.

    PubMed

    Frye, Cheryl A; Petralia, Sandra M; Rhodes, Madeline E; Stein, Benjamin

    2003-12-01

    5alpha-pregnan-3alpha-ol-20-one (3alpha,5alpha-THP) in the ventral tegmental area (VTA) mediates lordosis of rodents. If fluoxetine's effects on lordosis are mediated in part by midbrain 3alpha,5alpha-THP, then fluoxetine regimens that decrease and increase lordosis would be expected to respectively lower and elevate midbrain 3alpha,5-THP levels. Experiment 1: Ovariectomized (ovx) rats received estradiol benzoate (EB; 5 micro g, SC) at 0 and 24 h and fluoxetine (20 mg/kg, IP) or vehicle 30 min before sex testing and tissue collection. Other rats received fluoxetine (10 mg/kg, IP) or vehicle for 15 days followed by EB-priming and testing. Systemic acute or chronic fluoxetine significantly decreased lordosis and midbrain 3alpha,5alpha-THP levels compared to vehicle. Experiment 2: Ovx rats with unilateral cannula to the VTA were primed with EB (5 micro g; 0, 24 h) and/or progesterone (0 or 100 micro g; 44 h, SC). At 47.5 h, fluoxetine (3.6 mM) or vehicle was infused to the VTA. At 48 h, rats were tested. Administering fluoxetine to the VTA significantly increased lordosis and midbrain 3alpha,5alpha-THP levels compared to vehicle infusions. Experiment 3: Ovx EB-primed rats were tested prior to, and 30 min after, treatmemt with acute fluoxetine (20 mg/kg, IP). Rats were then infused with 3alpha,5alpha-THP (100 ng) or vehicle to the VTA and were retested. 3alpha,5alpha-THP, but not vehicle, to the VTA reversed acute fluoxetine's inhibitory effects on lordosis. Together, these data suggest fluoxetine may alter lordosis in part through actions of 3alpha-THP in the midbrain.

  8. Potential bile acid metabolites. 19. The epimeric 3 alpha,6,7 beta-trihydroxy- and 3 alpha,6,7 beta,12 alpha-tetrahydroxy-5 alpha-cholanoic acids.

    PubMed

    Iida, T; Nishida, S; Chang, F C; Niwa, T; Goto, J; Nambara, T

    1993-04-01

    Syntheses by a new procedure of the known 3 alpha,6 alpha,7 beta- and 3 alpha,6 beta,7 beta-trihydroxy-5 alpha-cholanoic acids, and of the once-reported analog 3 alpha,6 alpha,7 beta,12 alpha-, as well as the new 3 alpha,6 beta,7 beta,12 alpha-tetrahydroxy-5 alpha-cholanoic acids, are described. Key intermediates of the syntheses are the 6-oxo-7 beta-ols of the respective 5 alpha-cholanoic acids (and their methyl esters) prepared by allomerization at C-5 of appropriate 6-bromo-7-oxo derivatives of the corresponding 5 beta-acids. Successful reduction of the 6,7-ketols to the desired products depended on the proper choice of reagents, either Zn(BH4)2 or Li/NH3/MeOH.

  9. Contribution from 3 alpha-Condensed States to the Triple-Alpha Reaction

    SciTech Connect

    Kato, Kiyoshi; Kurokawa, Chie; Arai, Koji

    2010-06-01

    The alpha-condensed state in nuclear systems has been proposed by Tohsaki et al. and has given rise to interesting discussions. The Hoyle state of {sup 12}C has been studied as the most typical example of such an alpha-condensed state. A new resonant 0{sub 3}{sup +} state (E{sub r} = 1.66 MeV, GAMMA = 1.48 MeV) is predicted as an excited alpha-condensed state in addition to the second 0{sup +} state of the Hoyle state by calculations of the 3 alpha orthogonality condition model (3 alpha OCM) using the complex scaling method. Based on this result, the breakup strengths of the inversion reaction for sequential ({sup 8}Be+alpha->{sup 12}C+gamma) and direct (alpha+alpha+alpha->{sup 12}C+gamma) processes are calculated. It is discussed that a large reaction strength calculated recently by Ogata et al. in non-resonant energies is considered as a contribution from the excited 0{sub 3}{sup +} state.

  10. TOPO3alpha influences antigenic variation by monitoring expression-site-associated VSG switching in Trypanosoma brucei.

    PubMed

    Kim, Hee-Sook; Cross, George A M

    2010-07-08

    Homologous recombination (HR) mediates one of the major mechanisms of trypanosome antigenic variation by placing a different variant surface glycoprotein (VSG) gene under the control of the active expression site (ES). It is believed that the majority of VSG switching events occur by duplicative gene conversion, but only a few DNA repair genes that are central to HR have been assigned a role in this process. Gene conversion events that are associated with crossover are rarely seen in VSG switching, similar to mitotic HR. In other organisms, TOPO3alpha (Top3 in yeasts), a type IA topoisomerase, is part of a complex that is involved in the suppression of crossovers. We therefore asked whether a related mechanism might suppress VSG recombination. Using a set of reliable recombination and switching assays that could score individual switching mechanisms, we discovered that TOPO3alpha function is conserved in Trypanosoma brucei and that TOPO3alpha plays a critical role in antigenic switching. Switching frequency increased 10-40-fold in the absence of TOPO3alpha and this hyper-switching phenotype required RAD51. Moreover, the preference of 70-bp repeats for VSG recombination was mitigated, while homology regions elsewhere in ES were highly favored, in the absence of TOPO3alpha. Our data suggest that TOPO3alpha may remove undesirable recombination intermediates constantly arising between active and silent ESs, thereby balancing ES integrity against VSG recombination.

  11. Glycogen synthase kinase 3 alpha phosphorylates and regulates the osteogenic activity of Osterix.

    PubMed

    Li, Hongyan; Jeong, Hyung Min; Choi, You Hee; Lee, Sung Ho; Jeong, Hye Gwang; Jeong, Tae Cheon; Lee, Kwang Youl

    2013-05-10

    Osteoblast-specific transcription factor Osterix is a zinc-finger transcription factor that required for osteoblast differentiation and new bone formation. The function of Osterix can be modulated by post-translational modification. Glycogen synthase kinase 3 alpha (GSK3α) is a multifunctional serine/threonine protein kinase that plays a role in the Wnt signaling pathways and is implicated in the control of several regulatory proteins and transcription factors. In the present study, we investigated how GSK3α regulates Osterix during osteoblast differentiation. Wide type GSK3α up-regulated the protein level, protein stability and transcriptional activity of Osterix. These results suggest that GSK3α regulates osteogenic activity of Osterix.

  12. Detection and localization of Mip-3alpha/LARC/Exodus, a macrophage proinflammatory chemokine, and its CCR6 receptor in human pancreatic cancer.

    PubMed

    Kleeff, J; Kusama, T; Rossi, D L; Ishiwata, T; Maruyama, H; Friess, H; Büchler, M W; Zlotnik, A; Korc, M

    1999-05-17

    Macrophage Proinflammatory Human Chemokine-3alpha (Mip-3alpha/LARC/Exodus) belongs to a large family of chemotactic cytokines, which participate in directing inflammatory cell migration and in modulating angiogenesis. Mip-3alpha signals through a recently identified G-protein linked 7-transmembrane receptor, CCR6. In this study, we have characterized the expression of Mip-3alpha and CCR6 in 12 normal and 16 cancerous human pancreatic tissues and in 4 cultured pancreatic cancer cell lines, and assessed the effects of Mip-3alpha on growth and invasion of these cell lines. Pancreatic cancer tissues markedly overexpressed Mip-3alpha in comparison with normal pancreatic samples. By in situ hybridization Mip-3alpha and CCR6 mRNA moieties were present in cancer cells within the tumors. In addition, Mip-3alpha was abundant in the macrophages infiltrating the tumor mass. Mip-3alpha and its receptor CCR6 were expressed in all 4 tested pancreatic cancer cell lines. Mip-3alpha stimulated the growth of one cell line, enhanced the migration of another cell line, and was without effect in the other 2 cell lines. Together, our findings suggest that Mip-3alpha has the potential to act via autocrine and paracrine mechanisms to contribute to the pathobiology of human pancreatic cancer.

  13. Genomic organization of the CC chemokine mip-3alpha/CCL20/larc/exodus/SCYA20, showing gene structure, splice variants, and chromosome localization.

    PubMed

    Nelson, R T; Boyd, J; Gladue, R P; Paradis, T; Thomas, R; Cunningham, A C; Lira, P; Brissette, W H; Hayes, L; Hames, L M; Neote, K S; McColl, S R

    2001-04-01

    We describe the genomic organization of a recently identified CC chemokine, MIP3alpha/CCL20 (HGMW-approved symbol SCYA20). The MIP-3alpha/CCL20 gene was cloned and sequenced, revealing a four exon, three intron structure, and was localized by FISH analysis to 2q35-q36. Two distinct cDNAs were identified, encoding two forms of MIP-3alpha/CCL20, Ala MIP-3alpha/CCL20 and Ser MIP-3alpha/CCL20, that differ by one amino acid at the predicted signal peptide cleavage site. Examination of the sequence around the boundary of intron 1 and exon 2 showed that use of alternative splice acceptor sites could give rise to Ala MIP-3alpha/CCL20 or Ser MIP-3alpha/CCL20. Both forms of MIP-3alpha/CCL20 were chemically synthesized and tested for biological activity. Both flu antigen plus IL-2-activated CD4(+) and CD8(+) T lymphoblasts and cord blood-derived dendritic cells responded to Ser and Ala MIP-3alpha/CCL20. T lymphocytes exposed only to IL-2 responded inconsistently, while no response was detected in naive T lymphocytes, monocytes, or neutrophils. The biological activity of Ser MIP-3alpha/CCL20 and Ala MIP-3alpha/CCL20 and the tissue-specific preference of different splice acceptor sites are not yet known.

  14. Reversal of caffeine-induced anxiety by neurosteroid 3-alpha-hydroxy-5-alpha-pregnane-20-one in rats.

    PubMed

    Jain, N S; Hirani, K; Chopde, C T

    2005-04-01

    Caffeine has been shown to increase brain and plasma content of neurosteroid 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-THP) that allosterically modulates GABA(A) receptors. The present study evaluated the role of neurosteroid 3alpha,5alpha-THP in the caffeine-induced anxiogenic-like effect using the elevated plus-maze (EPM) test in rats. Acute administration of caffeine (50 or 100mg/kg, i.p.) produced anxiogenic-like activity that was reversed by pretreatment with the neurosteroid 3alpha,5alpha-THP or progesterone, the GABA(A) agonist muscimol, or the benzodiazepine receptor agonist diazepam. On the contrary, caffeine produced higher anxiety in animals previously treated with the GABA(A) receptor antagonist, bicuculline or either of the various neurosteroid biosynthesis enzyme inhibitors viz. trilostane, finasteride or indomethacin. Furthermore, pretreatment with DHEAS, a neurosteroid that negatively modulates GABA(A) receptors also enhanced the caffeine-induced anxiety. Moreover, adrenalectomy potentiated the anxiogenic-like response of caffeine indicating the contributory role of peripheral steroidogenesis. Thus, it is speculated that neurosteroid 3alpha,5alpha-THP through positive modulation of GABA(A) receptor activity may serve as a counter-regulatory mechanism against caffeine-induced anxiety.

  15. Aqueous chemical growth of alpha-Fe2O3-alpha-Cr203 nanocompositethin films

    SciTech Connect

    Vayssieres, Lionel; Guo, Jinghua; Nordgren, Joseph

    2001-06-30

    We are reporting here on the inexpensive fabrication and optical properties of an iron(III) oxide chromium(III) oxide nanocomposite thin film of corundum crystal structure. Its novel and unique-designed architecture consists of uniformed, well-defined and oriented nanorods of Hematite (alpha-Fe2O3) of 50 nm in diameter and 500nm in length and homogeneously distributed nonaggregated monodisperse spherical nanoparticles of Eskolaite (alpha-Cr2O3) of 250 nm in diameter. This alpha-Fe2O3 alpha-Cr2O3 nanocomposite thin film is obtained by growing, directly onto transparent polycrystalline conducting substrate, an oriented layer of hematite nanorods and growing subsequently, the eskolaite layer. The synthesis is carried out by a template-free, low-temperature, multilayer thin film coating process using aqueous solution of metal salts as precursors. Almost 100 percent of the light is absorbed by the composite film between 300 and 525 nm and 40 percent at 800 nm which yields great expectations as photoanode materials for photovoltaic cells and photocatalytic devices.

  16. GSK3 alpha and beta are new functionally relevant targets of tivantinib in lung cancer cells

    PubMed Central

    Remsing Rix, Lily L.; Kuenzi, Brent M.; Luo, Yunting; Remily-Wood, Elizabeth; Kinose, Fumi; Wright, Gabriela; Li, Jiannong; Koomen, John M.; Haura, Eric B.; Lawrence, Harshani R.; Rix, Uwe

    2014-01-01

    Tivantinib has been described as a potent and highly selective inhibitor of the receptor tyrosine kinase c-MET and is currently in advanced clinical development for several cancers including non-small cell lung cancer (NSCLC). However, recent studies suggest that tivantinib’s anticancer properties are unrelated to c-MET inhibition. Consistently, in determining tivantinib’s activity profile in a broad panel of NSCLC cell lines, we found that, in contrast to several more potent c-MET inhibitors, tivantinib reduces cell viability across most of these cell lines. Applying an unbiased, mass-spectrometry-based, chemical proteomics approach, we identified glycogen synthase kinase 3 (GSK3) alpha and beta as novel tivantinib targets. Subsequent validation showed that tivantinib displayed higher potency for GSK3α than for GSK3β and that pharmacological inhibition or simultaneous siRNA-mediated loss of GSK3α and GSK3β caused apoptosis. In summary, GSK3α and GSK3β are new kinase targets of tivantinib that play an important role in its cellular mechanism-of-action in NSCLC. PMID:24215125

  17. Calcium triggers beta-defensin (hBD-2 and hBD-3) and chemokine macrophage inflammatory protein-3 alpha (MIP-3alpha/CCL20) expression in monolayers of activated human keratinocytes.

    PubMed

    Pernet, I; Reymermier, C; Guezennec, A; Branka, J-E; Guesnet, J; Perrier, E; Dezutter-Dambuyant, C; Schmitt, D; Viac, J

    2003-12-01

    The inducible epidermal beta-defensins and the chemokine macrophage inflammatory protein-3alpha (MIP-3alpha/CCL20) are important mediators involved in innate and adaptive immunity and in the recruitment of immune cells. The aim of our study was to determine whether calcium could trigger the induction of beta-defensins (hBD-2 and hBD-3) mRNA and the release of MIP-3alpha by normal human keratinocyte monolayers. Epidermal cells derived from foreskin were cultured in defined medium supplemented with different calcium levels (0.09, 0.8 and 1.7 mM) and were stimulated or not with the pro-inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha 1-500 ng/ml) or interferon-gamma (INF-gamma 1-100 ng/ml). A high calcium concentration (1.7 mM) alone applied in culture medium for 4 days was sufficient to induce hBD-2 and hBD-3 mRNA expression. Whatever interindividual variability in the expression of hBD-2 and hBD-3 mRNA and MIP-3alpha secretion, the addition of TNF-alpha for a short duration (26h), initiated a dose-dependent and coordinated up-regulation of hBD-2 and hBD-3 mRNA and MIP-3alpha release in keratinocyte cultures. Unlike hBD-2 and hBD-3 mRNA was preferentially stimulated by IFN-gamma rather than TNF-alpha. In our experimental conditions, L-isoleucine, described to stimulate beta-defensin in bovine epithelial cells, did not exert any effect either on hBD-2 and hBD-3 transcripts or MIP-3alpha protein. Taken together, these results confirm the major role of the maturation/differentiation process of normal human keratinocytes in the induction of inducible beta-defensins and MIP-3alpha chemokine, which contribute in vivo to the immunosurveillance of the skin barrier function.

  18. Mechanisms regulating male sexual behavior in the rat: role of 3 alpha- and 3 beta-androstanediols.

    PubMed

    Morali, G; Oropeza, M V; Lemus, A E; Perez-Palacios, G

    1994-09-01

    To assess whether naturally occurring 5 alpha-androstanediols (5 alpha-androstane-3 alpha, 17 beta-diol and 5 alpha-androstane-3 beta, 17 beta-diol) play a role in the regulation of male sexual behavior in the rat, their capability to restore copulatory behavior in castrated animals was evaluated. Androstanediols were chronically administered either alone or in combination with 5 alpha-dihydrotestosterone (DHT) or with estradiol-17 beta (E2). Animals treated with testosterone (T), DHT, E2, and vehicle, either alone or in different combinations, served as controls. The occurrence of mounting, intromission, and ejaculation as well as detailed parameters of copulatory behavior were recorded twice per week for 3 weeks. At the end of treatments, the weights of sex accessory organs were also recorded. When 3 beta, 5 alpha-androstanediol (3 beta-diol; 500 micrograms/day) was administered in combination with DHT (300 micrograms/day), full copulatory behavior was restored in all subjects in a manner similar to that obtained with E2 plus DHT or T plus DHT combinations, thus indicating an estrogen-like behavioral effect of 3 beta-diol. Administration of 3 alpha, 5 alpha-androstanediol (3 alpha-diol; 500 micrograms/day) combined with DHT also restored sexual behavior, though to a lesser extent. When 3 alpha-diol (500 micrograms/day) was simultaneously administered with E2 (5 micrograms/day), the copulatory behavior of castrated animals was fully restored in a fashion similar to that observed after administration of DHT plus E2 and T plus E2 combinations, indicating a potent androgen-like effect of 3 alpha-diol.(ABSTRACT TRUNCATED AT 250 WORDS)

  19. Urinary analysis of 16(5alpha)-androsten-3alpha-ol by gas chromatography/combustion/isotope ratio mass spectrometry: implications in anti-doping analysis.

    PubMed

    Saudan, Christophe; Baume, Norbert; Mangin, Patrice; Saugy, Martial

    2004-10-15

    We present a method for the analysis of urinary 16(5alpha)-androsten-3alpha-ol together with 5beta-pregnane-3alpha,20alpha-diol and four testosterone metabolites: androsterone (Andro), etiocholanolone (Etio), 5alpha-androstane-3alpha,17beta-diol (5alphaA), 5beta-androstane-3alpha,17beta-diol (5betaA) by means of gas chromatography/combustion/isotopic ratio mass spectrometry (GC/C/IRMS). The within-assay and between-assay precision S.D.s of the investigated steroids were lower than 0.3 and 0.6 per thousand, respectively. A comparative study on a population composed of 20 subjects has shown that the differences of the intra-individual delta(13)C-values for 16(5alpha)-androsten-3alpha-ol and 5beta-pregnane-3alpha,20alpha-diol are less than 0.9 per thousand. Thereafter, the method has been applied in the frame of an excretion study following oral ingestion of 50 mg DHEA initially and oral ingestion of 50mg pregnenolone 48 h later. Our findings show that administration of DHEA does not affect the isotopic ratio values of 16(5alpha)-androsten-3alpha-ol and 5beta-pregnane-3alpha,20alpha-diol, whereas the isotopic ratio values of 5beta-pregnane-3alpha,20alpha-diol vary by more 5 per thousand upon ingestion of pregnenolone. We have observed delta(13)C-value changes lower than 1 per thousand for 16(5alpha)-androsten-3alpha-ol, though pregnenolone is a precursor of the 16-ene steroids. In contrast to 5beta-pregnane-3alpha,20alpha-diol, the 16-ene steroid may be used as an endogenous reference compound when pregnenolone is administered.

  20. Formation of varanic acid, 3 alpha, 7 alpha, 12 alpha, 24-tetrahydroxy-5 beta-cholestanoic acid from 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestanoic acid in Bombina orientalis.

    PubMed

    Une, M; Inoue, A; Hoshita, T

    1996-11-01

    Varanic acid (3 alpha, 7 alpha, 12 alpha, 24-tetrahydroxy-5 beta-cholestanoic acid; 24-OH-THCA) is almost the sole component of bile acids in the bile of Bombina orientalis. To examine in the mechanism of the formation of 24-OH-THCA, radiolabeled (25R)- and (25S)-3 alpha, 7 alpha, 12 alpha-trihdroxy-5 beta-cholestanoic acids [(25R)- and (25S)-THCA] and (24E)-3 alpha, 7 alpha, 12 alpha-trihdroxy-5 beta-cholest-24-enoic acid (delta 24-THCA) were administered intraperitoneally to B. orientalis, gallbladder bile was collected after 24 h, and bile acids were subsequently extracted. Then the bile acids were analyzed by means of radio thin-layer chromatography and radio high-performance liquid chromatography after conversion to p-bromophenacyl ester derivatives. Although delta 24-THCA was not converted to 24-OH-THCA, (25R)-THCA and (25S)-THCA were transformed to (24R,25R)-24-OH-THCA and (24R,25S)-24-OH-THCA, respectively. These results strongly suggest that 24-OH-THCA was transformed via direct hydroxylation of the saturated side chain of THCA, not via hydration to an alpha, beta-unsaturated acid, delta 24-THCA, in B. orientalis.

  1. Expression of the C-C chemokine MIP-3 alpha/CCL20 in human epidermis with impaired permeability barrier function.

    PubMed

    Schmuth, M; Neyer, S; Rainer, C; Grassegger, A; Fritsch, P; Romani, N; Heufler, C

    2002-04-01

    External assault to the skin is followed by an epidermal response including synthesis of DNA, lipids, cytokines and migration of antigen presenting cells. MIP-3 alpha (CCL20, LARC, Exodus-1, Scya20) is a recently described C-C chemokine, predominantly expressed in extralymphoid tissue, which is known to direct migration of dendritic cell precursors and memory lymphocytes to sites of antigen invasion. We assessed the expression of MIP-3 alpha in human skin using semi-quantitative polymerase chain reaction. In vivo, MIP-3 alpha mRNA was constitutively expressed at low levels in untreated human epidermis. After acute disruption of the epidermal permeability barrier MIP-3 alpha mRNA was upregulated in the epidermal fraction, whereas dermal MIP-3 alpha mRNA levels remained unchanged. In vitro, MIP-3 alpha was increased in cultured keratinocytes treated with IL-1 alpha and TNF-alpha and was present in immature and mature dendritic cells, THP-1 monocytic cells and activated T cells. Finally, skin biopsies from patients with psoriasis, contact dermatitis and mycosis fungoides showed abundant expression. In biopsies from atopic dermatitis and graft vs. host disease a weak signal was present, whereas no expression was found in scleroderma and toxic epidermal necrolysis. We conclude that regulation of MIP-3 alpha mRNA is part of the epidermal response to external assault. Its upregulation may represent a danger signal for increased immunosurveillance in barrier disrupted skin and inflammatory skin conditions with impaired barrier function to counteract potential antigen invasion.

  2. Overexpression of Reg3alpha increases cell growth and the levels of cyclin D1 and CDK4 in insulinoma cells.

    PubMed

    Cui, Wei; De Jesus, Kristine; Zhao, Hong; Takasawa, Shin; Shi, Bingyin; Srikant, Coimbatore B; Liu, Jun-Li

    2009-06-01

    Regenerating gene (Reg) family protein Reg3alpha is normally expressed in pancreatic acinar and endocrine cells. In order to explore its effect on islet beta-cell replication, insulinoma MIN6 cells were stably transfected with murine Reg3alpha cDNA. Determined using real-time PCR and Western blots, the levels of Reg3alpha mRNA and protein in Reg3alpha-transfected clones were increased 10- and 6-fold, respectively. Western blots also revealed that the protein was released into the culture medium, consistent with an endocrine effect. In MTT cell proliferation assay, Reg3alpha-overexpressing cells exhibited a 2-fold increase in the rate of cell growth. In order to investigate the intracellular mechanism, we studied cell cycle regulatory proteins. In Reg3alpha-expressing cells, we detected 2.2- and 2.5-fold increased levels of cyclin D1 and CDK4, respectively, which paralleled a 1.8-fold increase in the rate of Akt phosphorylation. It is established that beta-cell replication is associated with increased cyclin D1 and CDK4 levels; deficiency in CDK4 or cyclin D2 results in reduced beta-cell mass and diabetes. Our results suggest that Reg3alpha stimulates beta-cell replication, by activating Akt kinase and increasing the levels of cyclin D1/CDK4.

  3. Synthesis of a precursor for the preparation of 9 alpha,11 alpha-tritiated 5 alpha-androstane-3 alpha,17 beta-diol 17-glucuronide

    SciTech Connect

    Rao, P.N.; Damodaran, K.M.

    1984-03-01

    Starting from 11 beta-hydroxytestosterone, the synthesis of a strategic precursor, C-9 (11) unsaturated 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide (9a), for the preparation of 9 alpha,11 alpha-tritiated 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide has been achieved. The authors optimized the reaction conditions for catalytic reduction employing hydrogen and subsequent base hydrolysis followed by purification on Amberlite XAD-2 resin to obtain the saturated 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide.

  4. Synthesis of [3 alpha-3H]7-dehydrocholesterol via stable tritiated 4-phenyl-1,2,4-triazoline-3,5-dione derivative.

    PubMed

    Batta, A K; Salen, G; Tint, G S; Honda, A; Shefer, S

    1997-11-01

    Synthesis of [3 alpha-3H]7-dehydrocholesterol is described via protection of the 5,7-diene system in 7-dehydrocholesterol as the Diels-Alder adduct with 4-phenyl-1,2,4-triazoline-3,5-dione followed by oxidation of the hydroxyl group to give the 3-oxo adduct. Reduction of the keto adduct with [3H]sodium borohydride produced the adduct of [3 alpha-3H]7-dehydrocholesterol from which the radiolabeled sterol was obtained via treatment with lithium aluminum hydride. The advantage of the method is that highly labeled [3 alpha-3H]7-dehydrocholesterol can be prepared. Further, unlike 7-dehydrocholesterol, its adduct with 4-phenyl-1,2,4-triazoline-3,5-dione is stable and can be stored. This allows the preparation of small batches of [3 alpha-3H]7-dehydrocholesterol for immediate use in biological experiments, and losses due to decomposition of excess radiolabeled 7-dehydrocholesterol are minimized.

  5. Bile alcohol metabolism in man. Conversion of 5beta-cholestane-3alpha, 7alpha,12alpha, 25-tetrol to cholic acid.

    PubMed Central

    Salen, G; Shefer, S; Setoguchi, T; Mosbach, E H

    1975-01-01

    To study the role of C25-HYDROXY BILE ALCOHOLS AS PRECURSORS OF CHOlic acid, [G-3-H]5beta-cholestane-3alpha,7alpha12alpha,25-tetrol was administered intravenously to two subjects with cerebrotendinous xanthomatosis (CTX) and two normal individuals. One day after pulse labeling, radioactivity was present in the cholic acid isolated from the bile and feces of the subjects with CTX and the bile of the normal individuals. In the two normal subjects, the sp act decay curves of [G-3-H]-cholic acid were exponential, and no traces of [G-3-H]-5beta-cholestane-3alpha,7alpha,12alpha,25-tetrol were detected. In contrast, appreciable quantities of labeled 5beta-cholestane-3alpha,-7aopha,12alpha,25-tetrol were present in the bile and feces of the CTX subjects. The sp act vs. time curves of fecal [G-3-H]5beta-cholestane-3alpha,7alpha,12alpha,25-tetrol and [G-3-H]-cholic acid showed a precursor-product relationship. Although these results suggest that 5beta-cholestane-3alpha,7alpha,12alpha,25-tetrol may be a precursor of cholic acid in man, the possibility that C26-hydroxy intermediates represent the normal pathway can not be excluded. PMID:1141434

  6. Antimicrobial property of (+)-lyoniresinol-3alpha-O-beta-D-glucopyranoside isolated from the root bark of Lycium chinense Miller against human pathogenic microorganisms.

    PubMed

    Lee, Dong Gun; Jung, Hyun Jun; Woo, Eun-Rhan

    2005-09-01

    (+)-Lyoniresinol-3alpha-O-beta-D-glucopyranoside (1) was isolated from an ethyl acetate extract of the root bark from Lycium chinense Miller, and its structure was determined using 1D and 2D NMR spectroscopy including DEPT, HMQC, and HMBC. (+)-Lyoniresinol-3alpha-O-beta-D-glucopyranoside exhibited potent antimicrobial activity against antibiotic-resistant bacterial strains, methicillin-resistant Staphylococcus aureus (MRSA) isolated from patients, and human pathogenic fungi without having any hemolytic effect on human erythrocytes. In particular, compound 1 induced the accumulation of intracellular trehalose on C. albicans as stress response to the drug, and disrupted the dimorphic transition that forms pseudo-hyphae caused by the pathogenesis. This indicates that (+)-lyoniresinol-3alpha-O-beta-D-glucopyranoside has excellent potential as a lead compound for the development of antibiotic agents.

  7. Evaluation of a fluorometric-enzymatic method based on 3alpha-hydroxysteroid dehydrogenase for the mycotoxin zearalenone determination in corn.

    PubMed

    Andreu, Yolanda; Castillo, Juan R; de Marcos, Susana; Galbán, Javier

    2004-09-08

    A new method for the fluorometric determination of zearalenone (ZEN) based on its reaction with betaNADH in the presence of the enzyme 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD) is described. The procedure is based on the change in fluorescence intensity that takes place during the enzymatic reaction (excitation at 340nm and emission at 454nm). The optimum reaction conditions and the analytical characteristics were studied; linear response range (1-10mgl(-1)) and reproducibility (8mgl(-1), 2.7%, n=7). Moreover, a mathematical model explaining the analytical signal is proposed. The method has been applied to zearalenone determination in a spiked corn sample.

  8. Enzymatic synthesis of a selective inhibitor for alpha-glucosidases: alpha-acarviosinyl-(1-->9)-3-alpha-D-glucopyranosylpropen.

    PubMed

    Lee, Young-Su; Lee, Myoung-Hee; Lee, Hee-Seob; Lee, Seung-Jae; Kim, Young-Wan; Zhang, Ran; Withers, Stephen G; Kim, Kwan Soo; Lee, Sung-Joon; Park, Kwan-Hwa

    2008-07-09

    Here, we describe the enzymatic synthesis of novel inhibitors using acarviosine-glucose as a donor and 3-alpha-D-glucopyranosylpropen (alphaGP) as an acceptor. Maltogenic amylase from Thermus sp. (ThMA) catalyzed the transglycosylation of the acarviosine moiety to alphaGP. The two major reaction products were isolated using chromatographies. Structural analyses revealed that acarviosine was transferred to either C-7 or C-9 of the alphaGP, which correspond to C-4 and C-6 of glucose. Both inhibited rat intestine alpha-glucosidase competitively but displayed a mixed-type inhibition mode against human pancreatic alpha-amylase. The alpha-acarviosinyl-(1-->7)-3-alpha-D-glucopyranosylpropen showed weaker inhibition potency than acarbose against both alpha-glycosidases. In contrast, the alpha-acarviosinyl-(1-->9)-3-alpha-D-glucopyranosylpropen exhibited a 3.0-fold improved inhibition potency against rat intestine alpha-glucosidase with 0.3-fold inhibition potency against human pancreatic alpha-amylase relative to acarbose. In conclusion, alpha-acarviosinyl-(1-->9)-3-alpha-D-glucopyranosylpropen is a novel alpha-glucosidase-selective inhibitor with 10-fold enhanced selectivity toward alpha-glucosidase over alpha-amylase relative to acarbose, and it could be applied as a potent hypoglycemic agent.

  9. Three-dimensional structure of rat liver 3 alpha-hydroxysteroid/dihydrodiol dehydrogenase: a member of the aldo-keto reductase superfamily.

    PubMed Central

    Hoog, S S; Pawlowski, J E; Alzari, P M; Penning, T M; Lewis, M

    1994-01-01

    The 3.0-A-resolution x-ray structure of rat liver 3 alpha-hydroxysteroid dehydrogenase/dihydrodiol dehydrogenase (3 alpha-HSD, EC 1.1.1.50) was determined by molecular replacement using human placental aldose reductase as the search model. The protein folds into an alpha/beta or triose-phosphate isomerase barrel and lacks a canonical Rossmann fold for binding pyridine nucleotide. The structure contains a concentration of hydrophobic amino acids that lie in a cavity near the top of the barrel and that are presumed to be involved in binding hydrophobic substrates (steroids, prostaglandins, and polycyclic aromatic hydrocarbons) and inhibitors (nonsteroidal antiinflammatory drugs). At the distal end of this cavity lie three residues in close proximity that have been implicated in catalysis by site-directed mutagenesis--Tyr-55, Asp-50, and Lys-84. Tyr-55 is postulated to act as the general acid. 3 alpha-HSD shares significant sequence identity with other HSDs that belong to the aldo-keto reductase superfamily and these may show similar architecture. Other members of this family include prostaglandin F synthase and rho-crystallin. By contrast, 3 alpha-HSD shares no sequence identity with HSDs that are members of the short-chain alcohol dehydrogenase family but does contain the Tyr-Xaa-Xaa-Xaa-Lys consensus sequence implicated in catalysis in this family. In the 3 alpha-HSD structure these residues are on the periphery of the barrel and are unlikely to participate in catalysis. Images PMID:8146147

  10. Interaction between obesity and the Hypoxia Inducible Factor 3 Alpha Subunit rs3826795 polymorphism in relation with plasma alanine aminotransferase.

    PubMed

    Wang, Shuo; Song, Jieyun; Yang, Yide; Zhang, Yining; Chawla, Nitesh V; Ma, Jun; Wang, Haijun

    2017-07-28

    Hypoxia Inducible Factor 3 Alpha Subunit (HIF3A) DNA has been demonstrated to be associated with obesity in the methylation level, and it also has a Body Mass Index (BMI)-independent association with plasma alanine aminotransferase (ALT). However, the relation among obesity, plasma ALT, HIF3A polymorphism and methylation remains unclear. This study aims to identify the association between HIF3A polymorphism and plasma ALT, and further to determine whether the effect of HIF3A polymorphism on ALT could be modified by obesity or mediated by DNA methylation. The HIF3A rs3826795 polymorphism was genotyped in a case-control study including 2030 Chinese children aged 7-18 years (705 obese cases and 1325 non-obese controls). Furthermore, the HIF3A DNA methylation of the peripheral blood was measured in 110 severely obese children and 110 age- and gender- matched normal-weight controls. There was no overall association between the HIF3A rs3826795 polymorphism and ALT. A significant interaction between obesity and rs3826795 in relation with ALT was found (P inter = 0.042), with rs3826795 G-allele number elevating ALT significantly only in obese children (β' = 0.075, P = 0.037), but not in non-obese children (β' = -0.009, P = 0.741). Additionally, a mediation effect of HIF3A methylation was found in the association between the HIF3A rs3826795 polymorphism and ALT among obese children (β' = 0.242, P = 0.014). This is the first study to report the interaction between obesity and HIF3A gene in relation with ALT, and also to reveal a mediation effect among the HIF3A polymorphism, methylation and ALT. This study provides new evidence to the function of HIF3A gene, which would be helpful for future risk assessment and personalized treatment of liver diseases.

  11. Merozoite surface protein-3 alpha as a genetic marker for epidemiologic studies in Plasmodium vivax: a cautionary note

    PubMed Central

    2013-01-01

    Background Plasmodium vivax is the most widespread of the human malaria parasites in terms of geography, and is thought to present unique challenges to local efforts aimed at control and elimination. Parasite molecular markers can provide much needed data on P. vivax populations, but few such markers have been critically evaluated. One marker that has seen extensive use is the gene encoding merozoite surface protein 3-alpha (MSP-3α), a blood-stage antigen known to be highly variable among P. vivax isolates. Here, a sample of complete msp-3α gene sequences is analysed in order to assess its utility as a molecular marker for epidemiologic investigations. Methods Amplification, cloning and sequencing of additional P. vivax isolates from different geographic locations, including a set of Venezuelan field isolates (n = 10), yielded a sample of 48 complete msp-3α coding sequences. Characterization of standard population genetic measures of diversity, phylogenetic analysis, and tests for recombination were performed. This allowed comparisons to patterns inferred from the in silico simulation of a polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) protocol used widely. Results The larger sample of MSP-3α diversity revealed incongruence between the observed levels of nucleotide polymorphism, which were high in all populations, and the pattern of PCR-RFLP haplotype diversity. Indeed, PCR-RFLP haplotypes were not informative of a population’s genetic diversity and identical haplotypes could be produced from analogous bands in the commonly used protocol. Evidence of frequent and variable insertion-deletion mutations and recurrent recombination between MSP-3α haplotypes complicated the inference of genetic diversity patterns and reduced the phylogenetic signal. Conclusions The genetic diversity of P. vivax msp-3α involves intragenic recombination events. Whereas the high genetic diversity of msp-3α makes it a promising marker for some

  12. Prostate formation in a marsupial is mediated by the testicular androgen 5 alpha-androstane-3 alpha,17 beta-diol.

    PubMed

    Shaw, G; Renfree, M B; Leihy, M W; Shackleton, C H; Roitman, E; Wilson, J D

    2000-10-24

    Development of the male urogenital tract in mammals is mediated by testicular androgens. It has been tacitly assumed that testosterone acts through its intracellular metabolite dihydrotestosterone (DHT) to mediate this process, but levels of these androgens are not sexually dimorphic in plasma at the time of prostate development. Here we show that the 3 alpha-reduced derivative of DHT, 5 alpha-androstane-3 alpha,17 beta-diol (5 alpha-adiol), is formed in testes of tammar wallaby pouch young and is higher in male than in female plasma in this species during early sexual differentiation. Administration of 5 alpha-adiol caused formation of prostatic buds in female wallaby pouch young, and in tissue minces of urogenital sinus and urogenital tubercle radioactive 5 alpha-adiol was converted to DHT, suggesting that circulating 5 alpha-adiol acts through DHT in target tissues. We conclude that circulating 5 alpha-adiol is a key hormone in male development.

  13. Role of bran in normals. Serum levels of cholesterols, triglyceride, calcium and total 3 alpha-hydroxycholanic acid, and intestinal transit time.

    PubMed

    Winreich, J; Pedersen, O; Dinesen, K

    1977-01-01

    After the intake of approximately 24 g wheat bran daily for 5 weeks, 25 trainee nurses showed no changes in the serum levels of cholesterol, triglyceride, calcium or total 3 alpha-hydroxycholanic acid. On the other hand, the study revealed a reduced intestinal transit time with good correlation to an increased frequency of bowel movements. Average body weight fell significantly, by 0.4 kg. The daily caloric intake remained constant throughout the study period, whereas the calcium intake was significantly increased. Among the serum parameters and the dietary constituents, good correlation was found only between serum cholesterol and the dietary cholesterol content. In addition, an inverse relationship was demonstrated between the serum levels of cholesterol and total 3 alpha-hydroxycholanic acid. The significance of this observation is as yet unknown.

  14. Enzymic synthesis of di- and tri-saccharide glycosides, using glycosidases and beta-D-galactoside 3-alpha-sialyl-transferase.

    PubMed

    Nilsson, K G

    1989-06-01

    The following disaccharide glycosides were obtained in yields of 10-35% from the appropriate donor and acceptor glycosides by employing glycosidases as catalysts: alpha-D-Galp-(1----3)-alpha-D-GalpNAc-OEt (alpha-D-galactosidase), beta-D-Galp-(1----3)-alpha-D-GalpNAc-OEt and beta-D-Galp-(1----3)-beta-D-GalpNAc-OEtBr (beta-D-galactosidase), beta-D-GlcpNAc-(1----6)-beta-D-Galp-OMe and beta-D-GlcpNAc-(1----6)-alpha-D-Manp-OMe (beta-N-acetylglucosaminidase). With beta-D-GlcpNAc-OEtSiMe3 as the acceptor, beta-D-galactosidase gave beta-D-Galp-(1----3)-beta-D-GlcpNAc-OEtSiMe3 almost exclusively, whereas, with beta-D-GlcpNAc-OMe as the acceptor, beta-D-Galp-(1----3)-beta-D-GlcpNAc-OMe was formed in only slightly excess over teh analogous beta-(1----4)-linked glycoside. The use of beta-D-galactosidase and beta-D-galactoside 3-alpha-sialyltransferase in sequence provided a convenient route to the trisaccharide glycosides alpha-D-Neup5Ac-(2----3)-beta-D-Galp-(1----3)-alpha-D-GalpNAc-OEt, alpha-D-Neup5Ac-(2----3)-beta-D-Galp-(1----3)-beta-D-GalpNAc-OE tBr, and alpha-D-Neup5Ac-(2----3)-beta-D-Galp-(1----3)-beta-D-GlcpNAc-OMe.

  15. [Cloning of genes coding for 3-alpha-hydroxysteroid dehydrogenase and for (3-17)-beta-hydroxysteroid dehydrogenase from Pseudomonas testosteroni].

    PubMed

    Floch, H H; Abalain, J H; Di Stefano, S; Carré, J L; Abalain-Colloc, M L

    1995-01-01

    A genomic library of Pseudomonas testosteroi total DNA constructed from SauIIIA digests ligated to a lambda gt11 vector was probed with different polyclonal antibodies raised against purified 3 alpha-HSD and (3 beta-17 beta)-HSD. Two different clones reacting with one antibody were selected. The clone reacting with (3-17)beta-HSD antibody contained a 2,661-base pair insert and was found to contained an open reading frame of 765 base pair that corresponds to a protein of 254 amino-acid residues. A 1,492-base pair was inserte in pBR 322 plasmid vector; the recombinant bacterie over expressed the (3-17)beta-HSD gene. The clone reacting with 3 alpha-HSD antibody contained a 1746 base pair insert which contained an open reading frame of 696 base pairs that corresponds to a protein of 231 amino-acid residues. A search for homologous proteins was performed. Distant similarities were found between (3-17)beta-HSD and members of the short-chain alcool dehydrogenase (SCAD) family but no similarity was observed between 3 alpha-HSD and proteins of this family.

  16. Identification of noncollagenous sites encoding specific interactions and quaternary assembly of alpha 3 alpha 4 alpha 5(IV) collagen: implications for Alport gene therapy.

    PubMed

    Kang, Jeong Suk; Colon, Selene; Hellmark, Thomas; Sado, Yoshikazu; Hudson, Billy G; Borza, Dorin-Bogdan

    2008-12-12

    Defective assembly of alpha 3 alpha 4 alpha 5(IV) collagen in the glomerular basement membrane causes Alport syndrome, a hereditary glomerulonephritis progressing to end-stage kidney failure. Assembly of collagen IV chains into heterotrimeric molecules and networks is driven by their noncollagenous (NC1) domains, but the sites encoding the specificity of these interactions are not known. To identify the sites directing quaternary assembly of alpha 3 alpha 4 alpha 5(IV) collagen, correctly folded NC1 chimeras were produced, and their interactions with other NC1 monomers were evaluated. All alpha1/alpha 5 chimeras containing alpha 5 NC1 residues 188-227 replicated the ability of alpha 5 NC1 to bind to alpha3NC1 and co-assemble into NC1 hexamers. Conversely, substitution of alpha 5 NC1 residues 188-227 by alpha1NC1 abolished these quaternary interactions. The amino-terminal 58 residues of alpha3NC1 encoded binding to alpha 5 NC1, but this interaction was not sufficient for hexamer co-assembly. Because alpha 5 NC1 residues 188-227 are necessary and sufficient for assembly into alpha 3 alpha 4 alpha 5 NC1 hexamers, whereas the immunodominant alloantigenic sites of alpha 5 NC1 do not encode specific quaternary interactions, the findings provide a basis for the rational design of less immunogenic alpha 5(IV) collagen constructs for the gene therapy of X-linked Alport patients.

  17. Hypoxia-inducible factor (HIF)-3alpha is subject to extensive alternative splicing in human tissues and cancer cells and is regulated by HIF-1 but not HIF-2.

    PubMed

    Pasanen, Annika; Heikkilä, Minna; Rautavuoma, Kati; Hirsilä, Maija; Kivirikko, Kari I; Myllyharju, Johanna

    2010-07-01

    The hypoxia-inducible transcription factors (HIFs) play a central role in the response of cells to hypoxia. HIFs are alphabeta dimers, the human alpha subunit having three isoforms. HIF-3alpha is unique among the HIF-alpha isoforms in that its gene is subject to extensive alternative splicing. Database analyses have predicted the generation of six HIF-3alpha splice variants that utilize three alternative transcription initiation sites. None of these variants is likely to act as an efficient transcription factor, but some of them have been reported to inhibit HIF-1 and HIF-2 functions. We analyzed here for the first time in detail whether these six variants are indeed generated in various human tissues and cell lines. We identified four novel variants, named here HIF-3alpha7 to HIF-3alpha10, whereas we obtained no evidence for the predicted HIF-3alpha3 and HIF-3alpha5. Distinct differences in the expression patterns of the variants were found between human tissues, the levels being particularly low in many cancer cell lines. Hypoxia upregulated transcription from all three alternative HIF-3alpha promoters. siRNA experiments showed that this induction is mediated specifically by HIF-1 and not by HIF-2. The tissue-specific differences in the expression patterns and levels of the HIF-3alpha variants can be expected to modulate the hypoxia response of various tissues and cell types to different extents during development and in pathological situations. A further level of regulation is brought about by the fact that the levels of the HIF-3alpha transcripts themselves are regulated by hypoxia and by changes in HIF-1 levels. Copyright 2010 Elsevier Ltd. All rights reserved.

  18. Purification and properties of a 3 alpha-hydroxysteroid dehydrogenase of rat liver cytosol and its inhibition by anti-inflammatory drugs.

    PubMed Central

    Penning, T M; Mukharji, I; Barrows, S; Talalay, P

    1984-01-01

    An NAD(P)-dependent 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50) was purified to homogeneity from rat liver cytosol, where it is responsible for most if not all of the capacity for the oxidation of androsterone, 1-acenaphthenol and benzenedihydrodiol (trans-1,2-dihydroxycyclohexa-3,5-diene). The dehydrogenase has many properties (substrate specificity, pI, Mr, amino acid composition) in common with the dihydrodiol dehydrogenase (EC 1.3.1.20) purified from the same source [Vogel, Bentley, Platt & Oesch (1980) J. Biol. Chem. 255, 9621-9625]. Since 3 alpha-hydroxysteroids are by far the most efficient substrates, the enzyme is more appropriately designated a 3 alpha-hydroxysteroid dehydrogenase. It also promotes the NAD(P)H-dependent reductions of quinones (e.g. 9,10-phenanthrenequinone, 1,4-benzoquinone), aromatic aldehydes (4-nitrobenzaldehyde) and aromatic ketones (4-nitroacetophenone). The dehydrogenase is not inhibited by dicoumarol, disulfiram, hexobarbital or pyrazole. The mechanism of the powerful inhibition of this enzyme by both non-steroidal and steroidal anti-inflammatory drugs [Penning & Talalay (1983) Proc. Natl. Acad. Sci. U.S.A. 80, 4504-4508] was examined with several substrates. Most non-steroidal anti-inflammatory drugs are competitive inhibitors (e.g. Ki for indomethacin, 0.20 microM for 9,10-phenanthrenequinone reduction at pH 6.0, and 0.835 microM for androsterone oxidation at pH 7.0), except for salicylates, which act non-competitively (e.g. Ki for aspirin, 650 microM for androsterone oxidation). The inhibitory potency of these agents falls sharply as the pH is increased from 6 to 9. Most anti-inflammatory steroids are likewise competitive inhibitors, except for the most potent (betamethasone and dexamethasone), which act non-competitively. The enzyme is inhibited competitively by arachidonic acid and various prostaglandins. PMID:6435601

  19. A human-mouse chimera of the alpha3alpha4alpha5(IV) collagen protomer rescues the renal phenotype in Col4a3-/- Alport mice.

    PubMed

    Heidet, Laurence; Borza, Dorin-Bogdan; Jouin, Mélanie; Sich, Mireille; Mattei, Marie-Geneviève; Sado, Yoshikazu; Hudson, Billy G; Hastie, Nicholas; Antignac, Corinne; Gubler, Marie-Claire

    2003-10-01

    Collagen IV is a major structural component of basement membranes. In the glomerular basement membrane (GBM) of the kidney, the alpha3, alpha4, and alpha5(IV) collagen chains form a distinct network that is essential for the long-term stability of the glomerular filtration barrier, and is absent in most patients affected with Alport syndrome, a progressive inherited nephropathy associated with mutation in COL4A3, COL4A4, or COL4A5 genes. To investigate, in vivo, the regulation of the expression, assembly, and function of the alpha3alpha4alpha5(IV) protomer, we have generated a yeast artificial chromosome transgenic line of mice carrying the human COL4A3-COL4A4 locus. Transgenic mice expressed the human alpha3 and alpha4(IV) chains in a tissue-specific manner. In the kidney, when expressed onto a Col4a3(-/-) background, the human alpha3(IV) chain restored the expression of and co-assembled with the mouse alpha4 and alpha5(IV) chains specifically at sites where the human alpha3(IV) was expressed, demonstrating that the expression of all three chains is required for network assembly. The co-assembly of the human and mouse chains into a hybrid network in the GBM restores a functional GBM and rescues the Alport phenotype, providing further evidence that defective assembly of the alpha3-alpha4-alpha5(IV) protomer, caused by mutations in any of the three chains, is the pathogenic mechanism responsible for the disease. This line of mice, humanized for the alpha3(IV) collagen chain, will also provide a valuable model for studying the pathogenesis of Goodpasture syndrome, an autoimmune disease caused by antibodies against this chain.

  20. Studies on the receptors to 5alpha-androst-16-en-3-one and 5alpha-androst-16-en-3alpha-ol in sow nasal mucosa.

    PubMed

    Gennings, J N; Gower, D B; Bannister, L H

    1977-02-28

    The presence of receptors to the "boar taint" pheromones 5alpha-androst-16-en-3-one and 5alpha-androst-16-en-3alpha-ol has been demonstrated in sow olfactory mucosa. Binding studies indicated that a sufficiently low concentration of olfactory tissue homogenate exhibited saturation of binding of 5alpha-androst-16-en-3-one, and this was of high affinity compared with control tissues of non-olfactory and heated olfactory tissues. Analysis of receptor binding of 5alpha-androst-16-en-3-one gave a value for the affinity constant (Ka) of approx. 8.3-10(8) M-1 and the value for the molar concentration of binding sites (n[M]) was approx. 3.3 pmol/mg protein. Almost identical values of Ka and n [M] were obtained when receptor binding of 5alpha-[5alpha-3H]androst-16-en-3alpha-ol was investigated (Ka 8.4-10(8) M-1; n [M] 3.7 pmol/mg protein). This suggests that the same receptor binds both 5alpha-androst-16-en-3-one and 5alpha-androst-16-en-3alpha-ol with equally high affinity. In a preliminary investigation to establish the specificity of the receptor, the binding of 17beta-hydroxy-5alpha-androstan-3-one was assayed; this steroid is odourless but has a similar structure except in ring D to 5alpha-androst-16-en-3-one. Binding was of the low affinity, non-specific type only, indicating that the sow olfactory receptors are not sensitive to this androgen.

  1. Potential anti-tumor-promoting activity of 3alpha-hydroxy-D:A-friedooleanan-2-one from the stem bark of Mallotus philippensis.

    PubMed

    Tanaka, Reiko; Nakata, Tomoko; Yamaguchi, Chiharu; Wada, Shun-Ichi; Yamada, Takeshi; Tokuda, Harukuni

    2008-03-01

    Four known friedelane-type triterpenoids, friedelin ( 1), 3-hydroxy-D:A-friedoolean-3-en-2-one ( 2), 2beta-hydroxy-D:A-friedooleanan-3-one ( 3), and 3alpha-hydroxy-D:A-friedooleanan-2-one ( 4), and two known lupane-type triterpenoids, lupeol ( 5) and betulin ( 6), were isolated from the stem bark of Mallotus philippensis. Isolates 1 - 4 and their synthetic analogues, 3-acetoxy-D:A-friedoolean-3-en-2-one ( 2A) and 3alpha-acetoxy-D:A-friedooleanan-2-one ( 4A), were tested for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12- O-tetradecanoylphorbol 13-acetate (TPA). The inhibitory effect of compounds 2 (IC (50) = 292 mol ratio/32 pmol/TPA) and 4 (IC (50) = 288) was stronger than those of the other compounds tested and the positive control, curcumin (IC (50) = 343). Compound 4 strongly inhibited mouse skin tumor promotion in an IN VIVO two-stage carcinogenesis model. Studies have been conducted to identify the biologically active compounds extracted from the leaves, bark, and cones of trees that currently have no specific commercial use and are therefore treated as waste in the forestry industry.

  2. Regulation of CCR6 chemokine receptor expression and responsiveness to macrophage inflammatory protein-3alpha/CCL20 in human B cells.

    PubMed

    Krzysiek, R; Lefevre, E A; Bernard, J; Foussat, A; Galanaud, P; Louache, F; Richard, Y

    2000-10-01

    The regulation of CCR6 (chemokine receptor 6) expression during B-cell ontogeny and antigen-driven B-cell differentiation was analyzed. None of the CD34(+)Lin(-) hematopoietic stem cell progenitors or the CD34(+)CD19(+) (pro-B) or the CD19(+)CD10(+) (pre-B/immature B cells) B-cell progenitors expressed CCR6. CCR6 is acquired when CD10 is lost and B-cell progeny matures, entering into the surface immunoglobulin D(+) (sIgD(+)) mature B-cell pool. CCR6 is expressed by all bone marrow-, umbilical cord blood-, and peripheral blood-derived naive and/or memory B cells but is absent from germinal center (GC) B cells of secondary lymphoid organs. CCR6 is down-regulated after B-cell antigen receptor triggering and remains absent during differentiation into immunoglobulin-secreting plasma cells, whereas it is reacquired at the stage of post-GC memory B cells. Thus, within the B-cell compartment, CCR6 expression is restricted to functionally mature cells capable of responding to antigen challenge. In transmigration chemotactic assays, macrophage inflammatory protein (MIP)-3alpha/CC chemokine ligand 20 (CCL20) induced vigorous migration of B cells with differential chemotactic preference toward sIgD(-) memory B cells. These data suggest that restricted patterns of CCR6 expression and MIP-3alpha/CCL20 responsiveness are integral parts of the process of B-lineage maturation and antigen-driven B-cell differentiation.

  3. Phenotypic consequences of deletion of the gamma 3, alpha 5, or beta 3 subunit of the type A gamma-aminobutyric acid receptor in mice.

    PubMed

    Culiat, C T; Stubbs, L J; Montgomery, C S; Russell, L B; Rinchik, E M

    1994-03-29

    Three genes (Gabrg3, Gabra5, and Gabrb3) encoding the gamma 3, alpha 5, and beta 3 subunits of the type A gamma-aminobutyric acid receptor, respectively, are known to map near the pink-eyed dilution (p) locus in mouse chromosome 7. This region shares homology with a segment of human chromosome 15 that is implicated in Angelman syndrome, an inherited neurobehavioral disorder. By mapping Gabrg3 on a panel of p-locus deletions, we have determined that the order of genes within this cluster is centromere-p(D15S12h)-Gabrg3-Gabra5-Gabrb3-telom ere. Like Gabrb3, neither the Gabra5 nor Gabrg3 gene is functionally imprinted in adult mouse brain. Mice deleted for all three subunits die at birth with a cleft palate, although there are rare survivors (approximately 5%) that do not have a cleft palate but do exhibit a neurological abnormality characterized by tremor, jerky gait, and runtiness. We have previously suggested that deficiency of the beta 3 subunit may be responsible for the clefting defect. Most notably, however, in this report we describe mice carrying two overlapping, complementing p deletions that fail to express the gamma 3 transcript, as well as mice from another line that express neither the gamma 3 nor alpha 5 transcripts. Surprisingly, mice from both of these lines are phenotypically normal and do not exhibit any of the neurological symptoms characteristic of the rare survivors that are deleted for all three (gamma 3, alpha 5, and beta 3) subunits. These mice therefore provide a whole-organism type A gamma-aminobutyric-acid receptor background that is devoid of any receptor subtypes that normally contain the gamma 3 and/or alpha 5 subunits. The absence of an overt neurological phenotype in mice lacking the gamma 3 and/or alpha 5 subunits also suggests that mutations in these genes are unlikely to provide useful animal models for Angelman syndrome in humans.

  4. Interferon-resistant Daudi cells are deficient in interferon-alpha-induced ISGF3 alpha activation, but remain sensitive to the interferon-alpha-induced increase in ISGF3 gamma content.

    PubMed

    Dron, M; Tovey, M G

    1993-10-01

    Low levels of the transcription factor ISGF3 alpha were detected in the cytoplasm and nucleus of untreated Daudi cells, which increased markedly following interferon (IFN) treatment. In contrast no ISGF3 alpha was detected in an IFN-resistant clone of Daudi cells, DIF8, and only low levels were detected in these cells after IFN-alpha treatment. High levels of ISGF3 were produced in vitro, however, by the addition of ISGF3 alpha to extracts of IFN-treated DIF8 cells, indicating that IFN is unable to produce substantial amounts of functional ISGF3 alpha in DIF8 cells. A second clone of IFN-resistant Daudi cells, DIF3, also exhibited defective ISGF3 alpha production, which was restored to normal in the subclone DIF3REV5 that had reverted to high IFN sensitivity. Thus, the antiproliferative effect of IFN on Daudi cells and derived clones is closely related to the level of ISGF3 present in the nucleus of these cells. IFN-alpha, however, also enhances the content of ISGF3 gamma in IFN-resistant cells as well as certain proteins of unknown function, raising the possibility that a second pathway of IFN-alpha signal transduction, distinct from the ISGF3 pathway, remains functional in both DIF8 and DIF3 cells.

  5. Gonadotrophin-releasing hormone agonist stimulates milt fluidity and plasma concentrations of 17,20beta-dihydroxylated and 5beta-reduced, 3alpha-hydroxylated C21 steroids in male plaice (Pleuronectes platessa).

    PubMed

    Vermeirssen, E L; Scott, A P; Mylonas, C C; Zohar, Y

    1998-11-01

    Spermiating male plaice were caught in the North Sea and acclimatised to laboratory conditions. In two experiments, males were injected intramuscularly with either microspheres or pellets containing gonadotrophin-releasing hormone agonist (GnRHa). Blood was sampled at 2- to 5-day intervals. Individual blood plasma specimens were assayed for testosterone, 5beta-reduced, 3alpha-hydroxy ("5beta,3alpha") steroids and sulphated 17, 20beta-dihydroxy ("17,20beta") steroids. Pooled plasma samples were also assayed for free and sulphated 17, 20beta-dihydroxy-4-pregnen-3-one, free 11-ketotestosterone, and glucuronidated testosterone and 11-ketotestosterone. Plasma concentrations of all steroids were significantly elevated by GnRHa from 2 to 5 days onwards following treatment. The most marked changes occurred in the concentrations of the sulphated 17,20beta steroids, which comprised approximately equal amounts of 5beta-pregnane-3alpha,17,20beta-triol 20-sulphate (3alpha,17, 20beta-P-5beta-S) and 5beta-pregnane-3beta,17,20beta-triol 20-sulphate, rising from ca. 1 to 30-80 ng/ml in the first and from ca. 8 to 80 ng/ml in the second experiment. Concentrations of 5beta, 3alpha steroids matched those of 17,20beta steroids in one experiment. However, in the other experiment, the two RIAs yielded highly disparate results in about 50% of the fish (including males in the control group). The plasma of these fish contained excessive amounts of 5beta,3alpha-immunoreactive material between 10 and 25 days. This material was identified as 3alpha,17, 21-trihydroxy-5beta-pregnan-20-one 21-sulphate (a metabolite of 11-deoxycortisol). All previous studies have indicated that when plasma concentrations of this steroid are high, so are those of 3alpha,17,20beta-P-5beta-S. This is the first indication that these steroids are regulated independently. In a third experiment, milt fluidity and production were assessed at 10, 15, and 25 days following GnRHa implantation. Milt volume and fluidity were

  6. The selenium-75-homocholic acid taurine test reevaluated: combined measurement of fecal selenium-75 activity and 3 alpha-hydroxy bile acids in 211 patients

    SciTech Connect

    van Tilburg, A.J.; de Rooij, F.W.; van den Berg, J.W.; Kooij, P.P.; van Blankenstein, M. )

    1991-06-01

    The recommended reference values for the selenium-75-homocholic acid taurine (75SeHCAT) test, used in the analysis of chronic diarrhea, were evaluated in 211 patients by comparing simultaneous measurements of 3 alpha-hydroxy bile acids and 75Se activity in daily collected stools. An initial evaluation in 11 patients showed that the fecal collection method, which allows inspection and additional analysis of stools, was equivalent to the abdominal retention method. Selenium-75-HCAT whole-body retention half-life (WBR50) was greater than 2.8 days in less than 10% of the patients with bile acid malabsorption and less than 1.7 days in less than 10% of the normals. We recommend that a 75SeHCAT WBR50 less than 1.7 days is abnormal, a WBR50 greater than 2.8 days is normal, and a WBR50 in the range 1.7-2.8 days is equivocal, which was the case in 48% (94/195) of the patients in this study.

  7. Serum 5alpha-androstane-3alpha,17beta-diol, androsterone, and testosterone concentrations in the male rat. Influence of age and gonadotropin stimulation.

    PubMed

    Moger, W H

    1977-04-01

    Serum concentrations of testosterone plus dihydrotestosterone (T-DHT), 5 alpha-androstane-3 alpha, 17 beta-diol (Diol) and androsterone were measured during sexual maturation in male rats. Diol concentrations of 1 to 2.25 ng/ml were found in animals 10-90 days of age with no significant changes. Diol was the major androgen (5.75-8 times T-DHT) from age 20-40 days. Androsterone rose to 1.25 ng/ml at 25 days of age and declined to values of greater than 0.5 ng/ml from age 30-90 days. Testosterone-DHT levels were 1 ng/ml or less from 10-40 days of age, and then rose to a peak at 60 days. The ratio of Diol toT-DHT was significantly elevated from age 20-35 days, indicating that Diol is the major androgen in circulation at this time. Acute treatment of 33 day old rats with LH, but not with FSH, resulted in a dose-dependent increase in serum T-DHT, Diol, and androsterone. The dose-response and time course of response for the three steriods were nearly identical. Changes in testes capacity to secrete androgens were assessed 25, 33, 40 and 60 days of age by administering a maximum dose of LH before blood collection. Maximum response (sum of androsterone, Diol and T-DHT) occurred at 40 days with no further increase at 60 days.

  8. First and second transmembrane segments of alpha3, alpha4, beta2, and beta4 nicotinic acetylcholine receptor subunits influence the efficacy and potency of nicotine.

    PubMed

    Rush, Ray; Kuryatov, Alexander; Nelson, Mark E; Lindstrom, Jon

    2002-06-01

    The first three transmembrane segments (M1-M3) of human nicotinic acetylcholine receptors (nAChRs) have been implicated in determining the efficacy of nicotine by studies of alpha3/alpha4 subunit chimeras. Nicotine has full efficacy on the alpha4beta2 nAChR and partial efficacy on the alpha3beta2 nAChR. Now, we have exchanged individually three amino acids between the alpha4 and the alpha3 subunits at positions 226(M1), 258(M2), and 262(M2). Also, similar exchanges were made in the beta2 and beta4 subunits at positions 224(M1), 226(M1), and 254(M2) (using alpha subunit numbering). Expression of these mutated nAChRs in Xenopus laevis oocytes showed that the mutated M1 amino acids were important in influencing the potency of ACh and nicotine. It is hypothesized that these M1 amino acids affect the stability between the resting and activated states of the nAChR. M2 amino acids altered the efficacy of nicotine, usually without altering its potency. When the residue located at position 258 in the M2 region of the alpha subunit was valine (as in the alpha3 subunit), the resulting nAChR exhibited partial efficacy for nicotine that was voltage-dependent. Therefore, we believe that these M2 amino acids contribute to the formation of a binding site for nicotine in the alpha3beta2 nAChR channel, which results in a low-affinity channel block, causing the lower efficacy of nicotine on this nAChR.

  9. Factors responsible for the formation of different N-alkylated porphyrins in rat liver microsomal systems exposed to norethindrone. The role of 3 alpha-hydroxysteroid dehydrogenase.

    PubMed Central

    White, I N; Blakey, D C; Green, M L; Jarman, M; Schulten, H R

    1986-01-01

    Incubation of rat liver microsomes with norethindrone and a NADPH-generating system leads to the formation of one N-alkylated porphyrin (green pigment, GP1). Administration of this steroid to male rats in vivo results in the formation of three more-polar green pigments (GP2, 3 and 4). A cytosolic protein (green-pigment converting protein) has been purified from rat liver that, when added to liver microsomal mixtures containing norethindrone (0.03 mM) and a NADPH-generating system, results in the formation of all four green pigments (GP1, 2, 3 and 4). Field-desorption mass spectrometry of the purified green pigments gave protonated molecules, [M + H]+, at m/z 905 for GP1, m/z 909 for GP2, m/z 925 for GP3 and 4. The Mr of the purified cytosolic protein on SDS/polyacrylamide-gel electrophoresis or gel filtration was 37000. Polyacrylamide-gel isoelectric focusing gave a pI value of 5.9. Antibodies raised in rabbits against this protein, after preincubation with rat liver cytosol, subsequently prevented the formation of the more-polar norethindrone-induced green pigments (GP2, 3 and 4). The purified protein in the presence of either NADH or NADPH catalysed the reduction of delta 4-ring-reduced norethindrone, 5 alpha-oestran-17 alpha-ethynyl-17 beta-ol-3-one and, with the appropriate cofactor, the oxidation and reduction of steroids lacking the ethynyl function, e.g. androsterone or dihydrotestosterone. Indomethacin inhibited the reduction of dihydrotestosterone by this protein with an I50 (concn. causing 50% inhibition) value of 4.9 microM. From its physical and enzymic properties it is concluded that green-pigment converting protein is the same as 3 alpha-hydroxysteroid dehydrogenase (EC 1.1.1.50). Images Fig. 4. Fig. 7. PMID:3463301

  10. Molecular docking and QSAR studies on substituted acyl(thio)urea and thiadiazolo [2,3-alpha] pyrimidine derivatives as potent inhibitors of influenza virus neuraminidase.

    PubMed

    Sun, Jiaying; Cai, Shaoxi; Mei, Hu; Li, Jian; Yan, Ning; Wang, Qin; Lin, Zhihua; Huo, Danqun

    2010-09-01

    Surflex-Dock was employed to dock 36 thiourea and thiadiazolo [2,3-alpha] pyrimidine derivatives into neuraminidase 1a4g. Molecular docking results showed that hydrogen bonding, electrostatic, and hydrophobic features were important factors affecting inhibitory activities of these neuraminidase inhibitors. Moreover, there was a significant correlation between the predicted binding affinity (total scores) and experimental pIC50 values with correlation coefficient r=0.846 and p<0.0001. Hologram quantitative structure-activity relationship, comparative molecular field analysis, and comparative molecular similarity indices analysis were used to develop quantitative structure-activity relationship models. Squared multiple correlation coefficients (r2) of hologram quantitative structure-activity relationship, comparative molecular field analysis, and comparative molecular similarity indices analysis models were 0.899, 0.878, and 0.865, respectively. Squared cross-validated correlation coefficient (q2) of hologram quantitative structure-activity relationship, comparative molecular field analysis, and comparative molecular similarity indices analysis models was in turn 0.628, 0.656, and 0.509. In addition, squared multiple correlation coefficients for test set (r2test) of hologram quantitative structure-activity relationship, comparative molecular field analysis, and comparative molecular similarity indices analysis models were 0.558, 0.667, and 0.566, respectively. The most active sample ID 2 was taken as a template molecule to design new molecules. Based on the comparative molecular field analysis model, new compounds were designed by LeapFrog. Seven new compounds with improved binding energy and predicted activities were finally obtained.

  11. Role of S114 in the NADH-induced conformational change and catalysis of 3alpha-hydroxysteroid dehydrogenase/carbonyl reductase from Comamonas testosteroni.

    PubMed

    Chang, Yi-Hsun; Huang, Tzu-Jung; Chuang, Lea-Yea; Hwang, Chi-Ching

    2009-10-01

    3alpha-Hydroxysteroid dehydrogenase/carbonyl reductase reversibly catalyzes the oxidation of androsterone with NAD(+) to form androstanedione and NADH. In this study, we characterize the role of the conserved residue S114 in cofactor binding and catalysis, using site-directed mutagenesis, steady-state kinetics, fluorescence quenching and anisotropy measurements. The catalytic efficiency of V/K(NADH)Et for wild-type and S114A is 1.5 x10(7) and 3.8 x 10(3) M(-1) s(-1), respectively, suggesting that NADH association to wild-type and S114A mutant enzymes involves two steps, a bimolecular binding step and isomerization. The binding of NADH into a hydrophobic pocket in the active site of wild-type and S114A mutant enzymes restricts its motion and shields the fluorescence quenching from solvent, with an increase in the fluorescence intensity and a blue shift at the maximum wavelength. Furthermore, the binding of NADH leads to the protein fluorescence quenching, mainly due to fluorescence resonance energy transfer to NADH. S114A mutant enzyme decreases 3100-fold in V/Et with no apparent change in K(m) for substrates. Addition of NADH to S114A mutant enzyme induces a secondary structural change. These results suggest that S114 is important to maintain the correct conformation for the nucleotide binding and facilitate the reaction. Substitution of alanine for S114 eliminates the hydrogen bonding interaction with P185, causing a conformational change in a nonproductive binding of NADH and a significant loss of activity.

  12. Effects of the testosterone metabolite dihydrotestosterone and 5 alpha-androstan-3 alpha,17 beta-diol on very long chain fatty acid metabolism in X-adrenoleukodystrophic fibroblasts.

    PubMed

    Petroni, A; Blasevich, M; Uziel, G

    2003-08-08

    X-Adrenoleukodystrophy (X-ALD) is a peroxisomal disorder associated with the abnormal accumulation of very long chain fatty acids (VLCFA) in plasma and tissues. We have demonstrated that the androgen dihydrotestosterone (DHT) and 5 alpha-androstan-3 alpha,17 beta-diol (3 alpha-diol) have favorable effect on VLCFA metabolism. We have investigated the effect of androgens on peroxisomal beta-oxidation, the incorporation of labelled lignoceric acid into cholesterol esters and VLCFA elongation, in cultured skin-fibroblasts from control and X-ALD patients. The androgens significantly increased peroxisomal beta-oxidation in X-ALD fibroblasts although VLCFA levels were not normalized. The major effect was on the incorporation of labelled lignoceric acid into cholesterol esters, since the enhanced lignoceric acid incorporation into cholesterol ester fraction, which occurred in X-ALD fibroblasts, was reduced towards normal values. In contrast, the androgens had no effect on the elongation pathway.

  13. A new, major C27 biliary bile acid in the red-winged tinamou (Rhynchotus rufescens):25R-1beta, 3alpha,7alpha-trihydroxy-5beta-cholestan-27-oic acid.

    PubMed

    Hagey, Lee R; Kakiyama, Genta; Muto, Akina; Iida, Takashi; Mushiake, Kumiko; Goto, Takaaki; Mano, Nariyasu; Goto, Junichi; Oliveira, Cleida A; Hofmann, Alan F

    2009-04-01

    The chemical structures of the three major bile acids present in the gallbladder bile of the Red-winged tinamou (Rhynchotus rufescens), an early evolving, ground-living bird related to ratites, were determined. Bile acids were isolated by preparative reversed-phase HPLC. Two of the compounds were identified as the taurine N-acylamidates of 25R-3alpha,7alpha-dihydroxy-5beta-cholestan-27-oic acid (constituting 22% of biliary bile acids) and 25R-3alpha,7alpha,12alpha-trihydroxy-5beta-cholestan-27-oic acid (constituting 51%). The remaining compound, constituting 21% of biliary bile acids, was an unknown C27 bile acid. Its structure was elucidated by LC/ESI-MS/MS and NMR and shown to be the taurine conjugate of 25R-1beta, 3alpha, 7alpha-trihydroxy-5beta-cholestan-27-oic acid, a C27 trihydroxy bile acid not previously reported. Although C27 bile acids with a 1beta-hydroxyl group have been identified as trace bile acids in the alligator, this is the first report of a major biliary C27 bile acid possessing a 1beta-hydroxyl group.

  14. Polysaccharides in fungi. XXXVII. Immunomodulating activities of carboxymethylated derivatives of linear (1-->3)-alpha-D-glucans extracted from the fruiting bodies of Agrocybe cylindracea and Amanita muscaria.

    PubMed

    Yoshida, I; Kiho, T; Usui, S; Sakushima, M; Ukai, S

    1996-01-01

    Immunomodulating activities of three carboxymethylated derivatives (AG-AL-CMS, AG-AL-CMI, and AM-APP-CM) of linear (1-->3)-alpha-D-glucans from Agrocybe cylindracea and Amanita muscaria were evaluated with murine peritoneal macrophages playing an important role in tumor immunity. The ratio of macrophages in peritoneal exudate cells increased more than 50% after the administration of three carboxymethylated (1-->3)-alpha-D-glucans. These carboxymethylated (1-->3)-alpha-D-glucans exhibited higher potentiating activities for macrophages than carboxymethylated linear (1-->3)-beta-D-glucan (CMPS) in the potency of reduction of nitro blue tetrazolium, products of nitric oxide and the soluble cytotoxic factor, the amount of glucose consumption, and the activation of acid phosphatase. AG-AL-CMS, AG-AL-CMI, and AM-APP-CM were found to induce the tumor regressing factor in mouse serum, although the ability of the induction of this factor was weaker than that of CMPS. The reticuloendothelial system-potentiating activation of three carboxymethylated alpha-D-glucans was similar to that of the carboxymethylated beta-D-glucan. AG-AL-CMS and AG-AL-CMI, but not AM-APP-CM, were suggested to possess a higher-order structure, resulting from the formation of a fluorescent complex with aniline blue.

  15. Substrate specificity of THCA-CoA oxidases from rat liver light mitochondrial fractions on dehydrogenation of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid CoA thioester.

    PubMed

    Ikegawa, S; Goto, T; Mano, N; Goto, J

    1998-11-01

    The substrate specificity of rat liver peroxisomal 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoyl-CoA (THCA-CoA) oxidases, which catalyze the dehydrogenation of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) CoA thioester, having an asymmetric center at C-25, to form (24E)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholest-24-enoic acid (delta 24-THCA) CoA thioester, was studied. The stable isotope labeled substrates, [3,7,12-18O3]-(25R)- and (25S)-THCA CoA thioesters were synthesized by an exchange reaction of carbonyl oxygens on a steroid nucleus of 3,7,12-trioxo-5 beta-cholestanoic acid, followed by metal hydride reduction and condensation reaction with CoA. After incubation of a mixture of unlabeled (25R)- and 18O-labeled (25S)-THCA CoA thioester, or vice versa, with hepatic peroxisomal THCA-CoA oxidases, biotransformed delta 24-THCA was determined by liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. The delta 24-THCA was derived only from (25S)-THCA CoA thioester, indicating that the 25S epimer of THCA is a preferential substrate on dehydrogenation by THCA-CoA oxidases.

  16. Configuration at C-25 in 5 beta-cholestane-3 alpha,7 alpha,12 alpha,25,26-pentol excreted by patients with cerebrotendinous xanthomatosis: circular dichroism and 13C-NMR studies.

    PubMed

    Dayal, B; Salen, G; Toome, V; Tint, G S

    1986-12-01

    The configuration at C-25 in 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha,25,26-pentol isolated from the bile and feces of patients with cerebrotendinous xanthomtosis (CTX) was determined from the lanthanide-induced circular dichroism (CD) Cotton effects and 13C-NMR measurements. Under anhydrous conditions, CD spectra of 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha,25,26-pentol in the presence of Eu(fod)3 exhibited a large induced negative Cotton effect at 320 nm. On the basis of the empirical rule (primary-tertiary-alpha-diols) in which R compounds have positive Cotton effects and S compounds have negative Cotton effects at 320 nm, it was concluded that 25,26-pentol has the 1,2,glycol structure with C-25 having the S-configuration. This assignment was based upon comparison with model compounds, 25(R and S),26-dihydroxy cholesterols and 25(R and S),26-dihydroxy cholecalciferols whose single-crystal X-ray structure and 13C-NMR studies have been performed. It is suggested that these data may be helpful to clarify the stereospecificity of the hydroxylation of the terminal methyl group of the cholesterol side chain in CTX.

  17. The metabolism of 3alpha, 7alpha, 12alpha-trihydorxy-5beta-cholestan-26-oic acid in two siblings with cholestasis due to intrahepatic bile duct anomalies. An apparent inborn error of cholic acid synthesis.

    PubMed Central

    Hanson, R F; Isenberg, J N; Williams, G C; Hachey, D; Szczepanik, P; Klein, P D; Sharp, H L

    1975-01-01

    Studies were carried out in a family in which two children with cholestasis due to intrahepatic bile duct anomalies were shown to have increased amounts of the cholic acid precursor, 3alpha, 7alpha, 12alpha-trihydorxy-5beta-cholestan-26-oic acid (THCA). The metabolism of THCA was studied in one of these patients after an intravenous injection of (3H)THCA, and the cause of the increased amounts of THCA in this condition was found to be due to a metabolic defect in the conversion of this compound into cholic acid. A small amount of (3H)cholic acid was also identified after (3H)THCA administration, confirming that this metabolic defect was incomplete. Varanic acid (3alpha, 7alpha, 12alpha, 24xi-tetrahydorxy-5beta-cholestan-26-oic acid), a metabolite of THCA, could not be identified in either of these patients. By assuming that this compound would be conjugated and excreted if the metabolic block occurred after the formation of varanic acid, the defect in these patients appears to be due to a deficiency of a 24-hydroxylating enzyme system required to convert THCA into varanic acid. This condition appears to be transmitted in an autosomal recessive fashion, because the two affected patients were of opposite sex, and neither a normal sibling nor the two parents have increased amount of THCA in their bile. Images PMID:1159074

  18. Stereospecific dehydrogenation of (25R)- and (25S)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acids by acyl-CoA oxidase in rat liver light mitochondrial fraction.

    PubMed

    Ikegawa, S; Watanabe, H; Goto, T; Mano, N; Goto, J; Nambara, T

    1995-08-01

    From a stereochemical point of view, the dehydrogenation mechanism of the biotransformation of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) into (24E)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholest-24-enoic acid (delta 24-THCA) has been studied with capillary gas chromatography (GC)/negative ion chemical ionization (NICI)-mass spectrometry. After incubation of (24R,25R)- or (24S,25S)-[24,25-2H2]THCA, synthesized from (24E)-delta 24-THCA by a deuterated diimide reduction, with a rat liver light mitochondrial fraction, 5 beta-cholestanoic acids were extracted and derivatized into a pentafluorobenzyl (PFB) ester-dimethylethylsilyl (DMES) ether. Subsequent resolution into THCA and delta 24-THCA was attained by GC on a cross-linked 5% phenylmethyl silicone fused-silica capillary column monitored with a corresponding characteristic carboxylate anion [M-PFB]- in the NICI mode. The stereospecific elimination of a pro-R hydrogen at C-24 in both (25R)- and (25S)-THCA indicated syn-elimination for the former, whereas anti-elimination for the latter was observed.

  19. Human macrophage inflammatory protein-3alpha/CCL20/LARC/Exodus/SCYA20 is transcriptionally upregulated by tumor necrosis factor-alpha via a non-standard NF-kappaB site.

    PubMed

    Harant, H; Eldershaw, S A; Lindley, I J

    2001-12-14

    The 5'-flanking sequences of the human macrophage inflammatory protein-3alpha/CCL20 gene were cloned and transfected into G-361 human melanoma cells in a luciferase reporter construct. Tumor necrosis factor-alpha (TNF-alpha) treatment stimulated luciferase expression, and promoter truncations demonstrated that TNF-alpha inducibility is conferred by a region between nt -111 and -77, which contains a non-standard nuclear factor-kappaB (NF-kappaB) binding site. The requirement for NF-kappaB was demonstrated as follows: (i) mutations in this NF-kappaB site abrogated TNF-alpha responsiveness; (ii) TNF-alpha activated a construct containing two copies of the CCL20 NF-kappaB binding site; (iii) overexpression of NF-kappaB p65 activated the CCL20 promoter; (iv) NF-kappaB from nuclear extracts of TNF-alpha-stimulated cells bound specifically to this NF-kappaB site.

  20. Determination of 5alpha-androst-16-en-3alpha-ol in truffle fermentation broth by solid-phase extraction coupled with gas chromatography-flame ionization detector/electron impact mass spectrometry.

    PubMed

    Wang, Guan; Li, Yuan-Yuan; Li, Dong-Sheng; Tang, Ya-Jie

    2008-07-15

    A novel method using solid-phase extraction coupled with gas chromatography and flame ionization detector (FID)/electron impact mass spectrometry (EIMS) was developed for the determination of 5alpha-androst-16-en-3alpha-ol (androstenol), a steroidal compound belonging to the group of musk odorous 16-androstenes, in truffle fermentation broth. Comparison studies between FID and EIMS indicated two detectors gave similar quantitative results. The highest androstenol concentration of 123.5 ng/mL was detected in Tuber indicum fermentation broth, while no androstenol was found in Tuber aestivum fermentation broth. For the first time, this work confirmed the existence of androstenol in the truffle fermentation broth, which suggested truffle fermentation is a promising alternative for androstenol production on a large scale.

  1. Phenotypic consequences of deletion of the {gamma}{sub 3}, {alpha}{sub 5}, or {beta}{sub 3} subunit of the type A {gamma}-aminobutyric acid receptor in mice

    SciTech Connect

    Culia, C.T.; Stubbs, L.J.; Montgomery, C.S.; Russell, L.B.; Rinchik, E.M.

    1994-03-29

    Three genes (Gabrg3, Gabra5, and Gabrb3) encoding the {gamma}{sub 3}, {alpha}{sub 5}, and {beta}{sub 3} subunits of the type A {gamma}-aminobutyric acid receptor, respectively, are known to map near the pink-eyed dilution (p) locus in mouse chromosome 7. This region shares homology with a segment of human chromosome 15 that is implicated in Angelman syndrome, an inherited neurobehavioral disorder. By mapping Gabrg3-Gabra5-Gabrb3-telomere. Like Gabrb3, neither the Gabra5 nor Gabrg3 gene is functionally imprinted in adult mouse brain. Mice deleted for all three subunits die at birth with a cleft palate, although there are rare survivors ({approximately} 5%) that do not have a cleft palate but do exhibit a neurological abnormality characterized by tremor, jerky gait, and runtiness. The authors have previously suggested that deficiency of the {beta}{sub 3} subunit may be responsible for the clefting defect. Most notably, however, in this report they describe mice carrying two overlapping, complementing p deletions that fail to express the {gamma}{sub 3} transcript, as well as mice from another line that express neither the {gamma}{sub 3} nor {alpha}{sub 5} transcripts. Surprisingly, mice from both of these lines are phenotypically normal and do not exhibit any of the neurological symptoms characteristic of the rare survivors that are deleted for all three ({gamma}{sub 3}, {alpha}{sub 5}, and {beta}{sub 3}) subunits. These mice therefore provide a whole-organism type A {gamma}-aminobutyric-acid receptor background that is devoid of any receptor subtypes that normally contain the {gamma}{sub 3} and/or {alpha}{sub 5} subunits. The absence of an overt neurological phenotype in mice lacking the {gamma}{sub 3} and/or {alpha}{sub 5} subunits also suggests that mutations in these genes are unlikely to provide useful animal models for Angelman syndrome in humans.

  2. Five specificity patterns of (1----3)-alpha-L-fucosyltransferase activity defined by use of synthetic oligosaccharide acceptors. Differential expression of the enzymes during human embryonic development and in adult tissues.

    PubMed

    Mollicone, R; Candelier, J J; Mennesson, B; Couillin, P; Venot, A P; Oriol, R

    1992-04-10

    The use of synthetic trisaccharides as acceptors led to the definition of five main (1----3)-alpha-L-fucosyltransferase activity patterns in human adult tissues: (I). Myeliod cells, granulocytes, monocytes, and lymphoblasts, transfer an alpha-L-fucopyranosyl group to O-3 of a 2-acetamido-2-deoxy-D-glucosyl residue of H blood-group Type 2 oligosaccharide [alpha-L-Fucp-(1----2)-beta-D-Galp-(1----4)-beta-D-GlcpNAc----R] with Mn2+ as activator. (II) Brain has the same acceptor specificity pattern as myeloid cells, but can also use Co2+ as activator. (III) Plasma and liver transfer an alpha-L-furopyranosyl group to H blood-group Type 2 and to sialyl-N-acetyllactosamine [alpha-NeuAc-(2----3)-beta-D-Galp-(1----4)-beta-D-GlcpNAc----R]. (IV) Intestine, gall bladder, kidney, and milk have the same activity as (III), but also transfer an alpha-L-fucopyranosyl group to O-4 of a 2-acetamido-2-deoxy-D-glucose residue of H blood-group Type 1 [alpha-L-Fucp-(1----2)-beta-D-Galp-(1----3)-beta-D-GlcpNAc----R] and sialyl Type 1 [alpha-NeuAc-(1----3)-beta-D-Galp-(1----3)-beta-D-GlcpNAc----R]. (V) Stomach mucosa is not able to use sialyl-N-acetyllactosamine, but can transfer an alpha-L-fucopyranosyl group to the other Type 1 and Type 2 acceptors. Unlike in adult tissue, a single myeloid-like pattern of (1----3)-alpha-L-fucosyltransferase activity was found at early stages of development in all tissues tested. This embryonic enzyme is later progressively replaced by enzymes or mixtures of enzymes having the corresponding adult patterns of enzyme expression. All lymphoblastoid cell lines and half of the tumor epithelial cell lines tested expressed the myeloid-like pattern of enzyme found in normal embryonic tissues. The remaining tumor epithelial cell lines expressed different forms of (1----3/4)-alpha-L-fucosyltransferase acceptor specificity patterns.

  3. 3{alpha}-6{alpha}-Dihydroxy-7{alpha}-fluoro-5{beta}-cholanoate (UPF-680), physicochemical and physiological properties of a new fluorinated bile acid that prevents 17{alpha}-ethynyl-estradiol-induced cholestasis in rats

    SciTech Connect

    Clerici, Carlo . E-mail: clerici@unipg.it; Castellani, Danilo; Asciutti, Stefania; Pellicciari, Roberto; Setchell, Kenneth D.R. |; O'Connell, Nancy C. |; Sadeghpour, Bahman; Camaioni, Emidio; Fiorucci, Stefano; Renga, Barbara; Nardi, Elisabetta; Sabatino, Giuseppe; Clementi, Mattia; Giuliano, Vittorio; Baldoni, Monia; Orlandi, Stefano; Mazzocchi, Alessandro; Morelli, Antonio; Morelli, Olivia

    2006-07-15

    3{alpha}-6{alpha}-Dihydroxy-7{alpha}-fluoro-5{beta}-cholanoate (UPF-680), the 7{alpha}-fluorine analog of hyodeoxycholic acid (HDCA), was synthesized to improve bioavailability and stability of ursodeoxycholic acid (UDCA). Acute rat biliary fistula and chronic cholestasis induced by 17{alpha}-ethynyl-estradiol (17EE) models were used to study and compare the effects of UPF-680 (dose range 0.6-6.0 {mu}mol/kg min) with UDCA on bile flow, biliary bicarbonate (HCO{sub 3} {sup -}), lipid output, biliary bile acid composition, hepatic enzymes and organic anion pumps. In acute infusion, UPF-680 increased bile flow in a dose-related manner, by up to 40.9%. Biliary HCO{sub 3} {sup -} output was similarly increased. Changes were observed in phospholipid secretion only at the highest doses. Treatment with UDCA and UPF-680 reversed chronic cholestasis induced by 17EE; in this model, UDCA had no effect on bile flow in contrast to UPF-680, which significantly increased bile flow. With acute administration of UPF-680, the biliary bile acid pool became enriched with unconjugated and conjugated UPF-680 (71.7%) at the expense of endogenous cholic acid and muricholic isomers. With chronic administration of UPF-680 or UDCA, the main biliary bile acids were tauro conjugates, but modification of biliary bile acid pool was greater with UPF-680. UPF-680 increased the mRNA for cytochrome P450 7A1 (CYP7A1) and cytochrome P450 8B (CYP8B). Both UDCA and UPF-680 increased the mRNA for Na{sup +} taurocholate co-transporting polypeptide (NCTP). In conclusion, UPF-680 prevented 17EE-induced cholestasis and enriched the biliary bile acid pool with less detergent and cytotoxic bile acids. This novel fluorinated bile acid may have potential in the treatment of cholestatic liver disease.

  4. DPI-3290 [(+)-3-((alpha-R)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide]. I. A mixed opioid agonist with potent antinociceptive activity.

    PubMed

    Gengo, Peter J; Pettit, Hugh O; O'Neill, Scott J; Wei, Ke; McNutt, Robert; Bishop, Michael J; Chang, Kwen-Jen

    2003-12-01

    Compound (+)-3-((alpha-R)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide (DPI-3290), is one of a series of novel centrally acting agents with potent antinociceptive activity that binds specifically and with high affinity to opioid receptors. In saturation equilibrium binding studies performed at 25 degrees C using membranes from rat brain or guinea pig cerebellum, the Ki values measured for DPI-3290 at delta-, mu-, and kappa-opioid receptors were 0.18 +/- 0.02, 0.46 +/- 0.05, and 0.62 +/- 0.09 nM, respectively. In vas deferens isolated from laboratory mice, DPI-3290 decreased electrically induced tension development in a concentration-dependent manner with corresponding IC50 values of 1.0 +/- 0.3, 6.2 +/- 2.0, and 25.0 +/- 3.3 nM at delta-, mu-, and kappa-receptors, respectively. The activity of DPI-3290 in isolated vas deferens tissue was approximately 20,000, 175.8, and 1500 times more efficacious than morphine, and 492, 2.5, and 35 times more efficacious than fentanyl at delta-, mu-, and kappa-receptors, respectively. In ileal strips isolated from guinea pigs, DPI-3290 inhibited tension development with a corresponding IC50 value of 3.4 +/- 1.6 nM at mu-opioid receptors and 6.7 +/- 1.6 nM at kappa-opioid receptors. Intravenous administration of 0.05 +/- 0.007 mg/kg DPI-3290 produced a 50% antinociceptive response in rats. The antinociceptive properties of DPI-3290 were blocked by naloxone (0.5 mg/kg s.c.). Compared with morphine, this study demonstrated that DPI-3290 is more potent and elicited a similar magnitude of antinociceptive activity in the rat, actions mediated by its mixed opioid receptor agonist activity. The marked antinociceptive activity of DPI-3290 will likely provide a means for relieving severe pain in patients that require analgesic treatment.

  5. Chemical synthesis of (22E)-3alpha,6alpha,7alpha,12alpha-Tetrahydroxy-5beta-chol-22-en-24-oic acid and its N-acylamidated conjugates with glycine or taurine: precursors of the [22,23-(3)H] labelled tracers.

    PubMed

    Ogawa, Shoujiro; Adachi, Yuuki; Kakiyama, Genta; Shimada, Miki; Mano, Nariyasu; Goto, Junichi; Iida, Takashi

    2010-08-01

    (22E)-3alpha,6alpha,7alpha,12alpha-Tetrahydroxy-5beta-chol-22-en-24-oic acid and its N-acylamidated conjugates with glycine or taurine were synthesized from cholic acid. The key reactions employed are: 1) degradation of the side chain in intermediary C(24) 3alpha,6alpha,7alpha,12alpha-tetrahydroxylated bile acid to the corresponding C(22) 23,24-dinor-aldehyde, followed by Wittig reaction with methyl (triphenylphosphoranylidene)acetate and 2) N-acylamidation of the unconjugated tetrahydroxy-Delta(22)-5beta-cholenoic acid with glycine (or taurine) in the presence of diethylphosphorocyanide and triethylamine as coupling reagents.

  6. Reaction of (13S)-13-iodo-6beta-methoxy-3alpha,5-cyclo-13,14-seco-5alpha-androstane-14,17-dione with hydroxylamine and its application to the synthesis of new 13,14-seco steroids.

    PubMed

    Khripach, Vladimir A; Zhabinskii, Vladimir N; Kuchto, Anna I; Fando, Galina P; Zhiburtovich, Yuliya Y; Lyakhov, Alexander S; Govorova, Alla A; Groen, Marinus B; van der Louw, Jaap; de Groot, Aede

    2004-07-01

    The synthesis of 13,14-seco steroids starting from easily available (13S)-13-iodo-6beta-methoxy-3alpha,5-cyclo-13,14-seco-5alpha-androsta-14,17-dione is described. The C-17 ketone was converted regioselectively into its oxime with simultaneous stereoselective deiodination at C-13. The remaining C-14 carbonyl group was then reduced stereoselectively with Ca(BH4)2. The configurations at the relevant stereocenters of the thus obtained hydroxy oxime were determined by X-ray analysis. Successful regeneration of the C-17 carbonyl group was achieved by treatment of the corresponding oxime acetate with TiCl3.

  7. Assignment of the human genes for hepatocyte nuclear factor 3-{alpha}, -{beta}, and -{gamma} (HNF3A, HNF3B, HNF3G) to 14q12-q13, 20p11, and 19q13.2-q13.4

    SciTech Connect

    Mincheva, A.; Lichter, P.; Schuetz, G.; Kaestner, K.H.

    1997-02-01

    Studies of liver-specific gene expression have resulted in the characterization of several liver-enriched transcription factors, such as the hepatocyte nuclear factors (HNF)-1, -3, and -4. The members of the hepatocyte nuclear factor 3 family (HNF- 3) are known to play a role in the regulation of gene expression in liver and lung. Three HNF-3 proteins (HNF-3{alpha}, HNF-3{beta}, and HNF-3{gamma}) were identified in rat liver nuclear extracts by means of their binding activity with respect to promotor elements in the {alpha}{sub 1}-antitrypsin and transthyretin genes. Following purification of the HNF-3 DNA-binding activities from rat liver extracts, the corresponding cDNAs were cloned and sequenced. Each of the three binding activities was shown to be encoded by an individual gene. Furthermore, a high degree of sequence similarity was detected between the DNA-binding domains of the HNF-3 genes and the homeotic Drosophila melanogaster gene forkhead. Subsequently, sequences closely related to the HNF-3/forkhead DNA binding domain have been found in species ranging from yeast to human. Genomic fragments encoding the mouse HNF-3 genes as well as cDNAs have been cloned and characterized. The murine genes have been assigned the gene symbols Hnf3a, b, and g, respectively. In the mouse, Hnf3a and b, are expressed mainly in lung, liver, stomach, and small intestine, while Hnf3g is not expressed in lung but in ovary, testis, heart, and adipose tissue. 14 refs., 1 fig.

  8. Five different profiles of dihydropyridines in blocking T-type Ca(2+) channel subtypes (Ca(v)3.1 (alpha(1G)), Ca(v)3.2 (alpha(1H)), and Ca(v)3.3 (alpha(1I))) expressed in Xenopus oocytes.

    PubMed

    Furukawa, Taiji; Nukada, Toshihide; Namiki, Yoshiko; Miyashita, Yoriko; Hatsuno, Kento; Ueno, Yasunari; Yamakawa, Takeshi; Isshiki, Takaaki

    2009-06-24

    1,4-dihydropyridine (DHP) Ca(2+) antagonists have recently been shown to block T-type Ca(2+) channels, which may render favorable actions on cardiovascular systems. However, this evaluation remains to be done systematically for each T-type Ca(2+) channel subtype except for the Ca(v)3.1 (alpha(1G)) subtype. To address this issue at the molecular level, blocking effects of 14 kinds of DHPs (amlodipine, aranidipine, azelnidipine, barnidipine, benidipine, cilnidipine, efonidipine, felodipine, manidipine, nicardipine, nifedipine, nilvadipine, nimodipine, nitrendipine), which are clinically used for treatments of hypertension, on 3 subtypes of T-type Ca(2+) channels [Ca(v)3.2 (alpha(1H)), Ca(v)3.3 (alpha(1I)), and Ca(v)3.1 (alpha(1G))] were investigated in the Xenopus oocyte expression system using the two-microelectrode voltage-clamp technique. These 3 kinds (alpha(1H), alpha(1I) and alpha(1G)) of T-type channels were blocked by amlodipine, manidipine and nicardipine. On the other hand, azelnidipine, barnidipine, benidipine and efonidipine significantly blocked alpha(1H) and alpha(1G), but not alpha(1I) channels, while nilvadipine and nimodipine apparently blocked alpha(1H) and alpha(1I), but not alpha(1G) channels. Moreover, aranidipine blocked only alpha(1H) channels. By contrast, cilnidipine, felodipine, nifedipine and nitrendipine had little effects on these subtypes of T-type channels. The result indicates that the blockade of T-type Ca(2+) channels by derivatives of DHP Ca(2+) antagonist was selective for the channel subtype. Therefore, these selectivities of DHPs in blocking T-type Ca(2+) channel subtypes would provide useful pharmacological and clinical information on the mode of action of the drugs including side-effects and adverse effects.

  9. DPI-3290 [(+)-3-((alpha-R)-alpha-((2S,5R)-4-Allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide]. II. A mixed opioid agonist with potent antinociceptive activity and limited effects on respiratory function.

    PubMed

    Gengo, Peter J; Pettit, Hugh O; O'Neill, Scott J; Su, Ying Fu; McNutt, Robert; Chang, Kwen-Jen

    2003-12-01

    Allyl-2,5-dimethyl-1-piperazines have been of interest as analgesic agents for the management of moderate-to-severe pain. In this study, we compared the antinociceptive properties and respiratory depressant activity of one such agent, (+)-3-((alpha-R)-alpha-((2S,5R)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxybenzyl)-N-(3-fluorophenyl)-N-methylbenzamide (DPI-3290), with those of established narcotic analgesics, morphine and fentanyl. Intravenous administration of DPI-3290 in conscious laboratory rats increased antinociception in a dose-dependent manner with a corresponding ED(50) value of 0.05 +/- 0.0072 mg/kg. Simultaneous measurement of arterial blood gas in animals treated with DPI-3290 demonstrated dose-dependent increases in pCO2 with an ED(50) value of 0.91 +/- 0.22 mg/kg. In comparison, morphine and fentanyl increased antinociception in rats with ED(50) values of 2.01 +/- 0.0005 and 0.0034 +/- 0.00024 mg/kg, respectively, and the ED(50) value for morphine-induced changes in pCO2 was 4.23 +/- 0.72 mg/kg, whereas the ED(50) value for fentanyl-induced changes in pCO2 was 0.0127 +/- 0.0035 mg/kg. A separate series of experiments were designed to examine the effects of DPI-3290 on mu-opioid receptor induced antinociception and hypercapnia. Intravenous bolus doses of DPI-3290 that ranged from 0.2 to 1.0 mg/kg had no effect on antinociception mediated by alfentanil (2 microg/kg/min i.v.) but reduced hypercapnia by approximately 50%. Results from these studies demonstrate the equivalent antinociceptive efficacy of DPI-3290, morphine, and fentanyl but dramatic differences in the hypercapnia that antinociceptive doses of these drugs produce. When measured simultaneously, DPI-3290 had an 18.2-fold difference in the ratio comparing the ED(50) value for antinociception with the ED(50) value for changes in pCO2; this ratio was 2.1 for morphine and 3.7 for fentanyl. Furthermore, DPI-3290 reduced the alfentanil-mediated hypercapnia without any effect on antinociception

  10. Characterization of the anxiolytic properties of a novel neuroactive steroid, Co 2-6749 (GMA-839; WAY-141839; 3alpha, 21-dihydroxy-3beta-trifluoromethyl-19-nor-5beta-pregnan-20-one), a selective modulator of gamma-aminobutyric acid(A) receptors.

    PubMed

    Vanover, K E; Rosenzweig-Lipson, S; Hawkinson, J E; Lan, N C; Belluzzi, J D; Stein, L; Barrett, J E; Wood, P L; Carter, R B

    2000-10-01

    The purpose of this study was to evaluate the effects of a novel neuroactive steroid, Co 2-6749 (GMA-839; WAY-141839; 3alpha, 21-dihydroxy-3beta-trifluoromethyl-19-nor-5beta-pregnan-20-one), on gamma-aminobutyric acid(A) receptors in vitro and to define its anxiolytic-like effects and side effect profile in vivo. Co 2-6749 fully inhibited [(35)S]t-butylbicyclophosphorothionate binding in rat brain cortical membranes with an IC(50) value of 230 nM and in human gamma-aminobutyric acid(A) receptor subunit combinations of alpha1beta2gamma2L, alpha2beta2gamma2L, alpha3beta2gamma2L, alpha4beta3gamma2L, alpha5beta2gamma2L, and alpha6beta3gamma2L receptors (IC(50) values of 200, 200, 96, 2300, 210, and 2000 nM). Rats were trained in a Geller-Seifter operant conflict paradigm. Co 2-6749 caused a dose-related increase in punished responding with a minimum effective dose of 1.6 mg/kg, p.o., a wide therapeutic index relative to a decrease in unpunished responding and relative to ataxia, and no tolerance. Additionally, ethanol caused less than a 2-fold shift to the left in the dose-response function of Co 2-6749 in the rotorod procedure in rats. In a pigeon conflict paradigm, punished responding was maximally increased to 784% of vehicle control by 30 mg/kg, p.o., with a 2-h duration and no effect on unpunished responding at this dose. Similarly, punished responding in squirrel monkeys was maximally increased to 1774% of control by 10 mg/kg, p.o., with no effect on unpunished responding at this dose. With robust anxiolytic-like activity across species, a large separation between anxiolytic-like effects and sedation/ataxia, a minimal interaction with ethanol, a lack of tolerance, and apparent oral bioavailability, Co 2-6749 makes an ideal candidate for development as a novel anxiolytic drug.

  11. [Temperature effect correction for Chang'E-3 alpha particle X-ray spectrometer].

    PubMed

    Wu, Ming-Ye; Wang, Huan-Yu; Peng, Wen-Xi; Zhang, Cheng-Mo; Zhang, Jia-Yu; Cui, Xing-Zhu; Liang, Xiao-Hua; Wang, Jin-Zhou; Yang, Jia-Wei; Fan, Rui-Rui; Liu, Ya-Qing; Dong, Yi-Fan; Wu, Feng; Zhao, Xiao-Yun

    2012-07-01

    Alpha particle X-ray spectrometer (APXS) is one of the payloads of Chang'E-3 lunar rover of China's Lunar Exploration Project. The present paper introduces briefly the components of APXS, how it works and its working environment on the lunar surface. The environmental temperature effect has been studied with simulations and experiments, and the results show that the temperature of the APXS sensor will be varying during the measuring on the lunar surface. And another experiment reveals that the energy resolution becomes worse if the sensor's temperature is varying. In this paper, a correction method based on Pearson's chi-squared test is presented. The method can improve the energy resolution when the sensor's temperature is varying. We have tested the method with the spectra acquired by APXS in the temperature varying period of Temperature Cycling Test, and the results show that the method is efficient and reliable.

  12. [Near infrared distance sensing method for Chang'e-3 alpha particle X-ray spectrometer].

    PubMed

    Liang, Xiao-Hua; Wu, Ming-Ye; Wang, Huan-Yu; Peng, Wen-Xi; Zhang, Cheng-Mo; Cui, Xing-Zhu; Wang, Jin-Zhou; Zhang, Jia-Yu; Yang, Jia-Wei; Fan, Rui-Rui; Gao, Min; Liu, Ya-Qing; Zhang, Fei; Dong, Yi-Fan; Guo, Dong-Ya

    2013-05-01

    Alpha particle X-ray spectrometer (APXS) is one of the payloads of Chang'E-3 lunar rover, the scientific objective of which is in-situ observation and off-line analysis of lunar regolith and rock. Distance measurement is one of the important functions for APXS to perform effective detection on the moon. The present paper will first give a brief introduction to APXS, and then analyze the specific requirements and constraints to realize distance measurement, at last present a new near infrared distance sensing algorithm by using the inflection point of response curve. The theoretical analysis and the experiment results verify the feasibility of this algorithm. Although the theoretical analysis shows that this method is not sensitive to the operating temperature and reflectance of the lunar surface, the solar infrared radiant intensity may make photosensor saturation. The solutions are reducing the gain of device and avoiding direct exposure to sun light.

  13. Functional bread with n-3 alpha linolenic acid from whole chia (Salvia hispanica L.) flour.

    PubMed

    Luna Pizarro, Patricia; Almeida, Eveline Lopes; Coelho, Alessandra Silva; Sammán, Norma Cristina; Hubinger, Miriam Dupas; Chang, Yoon Kil

    2015-07-01

    This work proposed to study the effects of the addition of whole chia flour (WCF) on the technological, nutritional and sensory qualities of bread. Different WCF contents (0 and 20 %) and vital gluten (VG) (0 and 4 %) were added to bread according to a 2(2) central composite rotational design. WCF decreased the specific volume, lightness and hue angle of the bread loaves, but did not affect the chroma values. WCF and VG contributed to maintenance of the moisture content of the loaves during the storage period. The increased firmness found with the addition of high levels of WCF (more than 10 %) was countered by larger amounts of VG (more than 2 %). The optimum loaf (10 % WCF and 2 % VG) showed 26 % more lipids, 19 % more protein and 11 % more ash than the standard loaf (0 % WCF and 0 % VG). A better lipid profile was also found (higher omega-3 fatty acid content and a better omega-6/omega-3 ratio). Both breads were positively rated in the sensory profile analysis.

  14. ENEA extremity dosemeter based on LiF(Mg,Cu,P) to evaluate Hp(3,alpha).

    PubMed

    Mariotti, F; Fantuzzi, E; Morelli, B; Gualdrini, G; Botta, M C; Uleri, G; Bordy, J M; Denoziere, M

    2011-03-01

    Recent epidemiological studies suggest a rather low-dose threshold (<0.5 Gy) for the induction of a cataract of the eye lens. Some other studies even assume that there is no threshold at all. Therefore, protection measures have to be optimised and current dose limits for the eye lens may be reduced in the future. ICRP Publication 103 on H(p)(d), in §(136), reads that '… a depth d = 3 mm has been proposed for the rare case of monitoring the dose to the lens of the eye. In practice, however, H(p)(3) has rarely been monitored and H(p)(0.07) can be used for the same monitoring purpose… '. As recommended on the EU 'Technical recommendations for monitoring individuals occupationally exposed to external radiation', a test on the ENEA TL extremity dosemeter is herein reported. The results within the actual EU founded Optimization of RAdiation protection for MEDical staff (ORAMED) Project, whose WP2 is aimed at the quantity H(p)(3) and eye lens dosimetry in practice, are taken into account. The paper summarises the main aspects of the study carried out at ENEA-Radiation Protection Institute (Bologna, Italy) to provide practical solutions (in the use and the design) to evaluate the response of the ENEA TL extremity dosemeter in terms of H(p)(3).

  15. Nonlocal {alpha}{alpha} potential for {sup 12}C as a 3{alpha} system

    SciTech Connect

    Orabi, M.; Suzuki, Y.; Fujiwara, Y.

    2008-04-29

    Cluster-cluster local potentials reproducing the phase shifts of binary systems are usually adopted to describe few cluster sytems for their simplicity, but it turns out that the nonlocal nature of these composite clusters is something that we cannot discard.

  16. Radiopharmaceutical chemistry of targeted radiotherapeutics, Part 3: alpha-particle-induced radiolytic effects on the chemical behavior of (211)At.

    PubMed

    Pozzi, Oscar R; Zalutsky, Michael R

    2007-07-01

    Two characteristics of alpha-particles that enhance their potential for targeted radiotherapy are their high energy and approximately cellular range. Unfortunately, these properties also can have negative consequences, confounding the production of clinically relevant levels of radiopharmaceutical because of radiolytic effects. The purpose of this study was to evaluate the effect of radiation dose on the astatine species present before initiation of a labeling reaction and the potential role of these molecules in the efficiency of N-succinimidyl 3-(211)At-astatobenzoate (SAB) synthesis. The ranges of radiation dose evaluated were selected to reflect those that might be encountered in SAB synthesis for the preparation of clinical doses of (211)At-labeled radiopharmaceuticals. The distribution of astatine species present in methanol, and the yields for the synthesis of SAB from N-succinimidyl 3-(tri-n-butylstannyl)benzoate as a function of radiation dose, were determined by high-performance liquid chromatography. Radiation doses in the range of 500-12,000 Gy were evaluated using different (211)At time-activity combinations, and the effect of acetic acid, a normal component of astatodestannylation reactions, also was studied. Finally, the effect of the reducing agent sodium sulfite also was evaluated to characterize the nature of the species produced by radiolysis. At radiation doses below 1,000 Gy, high-performance liquid chromatography analysis indicated that more than 90% of the (211)At was present in methanol as a single species, At(1), whereas at higher doses, a second peak, At(2), emerged. At(1) decreased and At(2) increased in a radiation dose-dependent fashion, with At(2) becoming the predominant species at about 3,000 Gy. At(2) was identified as a reduced form of astatine, presumably astatide, which could not be efficiently oxidized to a species suitable for electrophilic astatination. In methanol/acetic acid, more than 95% of the astatine was present as At(2) even at doses below 1,400 Gy. The emergence of a reduced form of astatine, At(2), at higher radiation doses is consistent with the decline in SAB yields observed under these conditions. Alteration of the chemical form of the astatine by radiolysis could account for the declining yields noted in the preparation of clinical-level (211)At-labeled radiopharmaceuticals and when the labeling chemistry is initiated hours after (211)At production.

  17. Developmental expression of Ca(v)1.3 (alpha1d) calcium channels in the mouse inner ear.

    PubMed

    Hafidi, A; Dulon, D

    2004-06-21

    Voltage-gated calcium channels are important for neurotransmission at the level of inner hair cells (IHCs) and outer hair cells (OHCs). These channels open when mechanical stimulation depolarises the hair cell membrane and the resulting calcium influx triggers neurotransmitter release. Voltage-gated calcium channels expressed in hair cells are known to be of the L-type with a predominance of the Ca(v)1.3 subunit. The present study describes the developmental expression of the Ca(v)1.3 protein in the cochlea and the vestibular system using immunohistochemical technique. In the adult organ of Corti (OC), Ca(v)1.3 was localized in both sensory and non-sensory cells with a more intense expression in IHCs and Deiters cells when compared to OHCs. In both hair cell types, immunoreactivity was observed in the apical pole, basolateral membrane and at the basal pole (synaptic zone). Similar results were obtained in the vestibular organs. During development, Ca(v)1.3 immunoreactivity was observed in the cochlea as early as embryonic day 15, with expression increasing at birth. At these early stages of cochlear development, Ca(v)1.3 was expressed in all cell types surrounding the scala media. In the OC, the labeling was observed in IHCs, OHCs and supporting cells. The Ca(v)1.3 expression reached an adult-like pattern by the end of the second postnatal week. The present findings suggested that, in addition to their implication in hair cells synaptic transmission, Ca(v)1.3 calcium channels also play an important role in vesicle recycling and transport, as suggested by their extrasynaptic location at the apical pole of the hair cells. The Ca(v)1.3 channels in Deiters cells could participate in active calcium-induced changes in micromechanics of these supporting cells. An early expression during development suggested that these calcium channels are in addition important in the development of the cochlear and vestibular sensory epithelium.

  18. Reduction of 3 alpha-hydroxy-5 beta-chol-6-en-24-oic acid to lithocholic acid in rats

    SciTech Connect

    Kimura, K.; Ogura, M.

    1988-03-01

    After (24-/sup 14/C)delta 6-lithocholic acid was injected into the cecum of rats, (/sup 14/C)lithocholic acid was identified as a metabolite in feces. When the labeled delta 6-bile acid was injected intraperitoneally into bile-fistula rats, radioactivity excreted in bile was contained most abundantly in the taurine-conjugated fraction of bile acids. In the fraction, taurine conjugate of (/sup 14/C)delta 6-lithocholic acid but of neither (/sup 14/C)lithocholic acid nor other bile acids was found. The results showed that (24-/sup 14/C)delta 6-lithocholic acid was reduced to (/sup 14/C)lithocholic acid by the intestinal flora but not by the liver, which, however, was capable of conjugating delta 6-lithocholic acid with taurine.

  19. 4 Birds 1 Stone to Inhibit 5androstane-3alpha,17beta-diol Conversion to DHT

    DTIC Science & Technology

    2016-09-01

    culture models; and (B) Determine whether arylquins inhibit prostate cancer metastasis to the lung and other tissues and the growth of prostate cancer ...Unlimited 13. SUPPLEMENTARY NOTES 14. ABSTRACT Prostate cancer growth and survival relies on the interaction between androgen receptor and the...testicular androgens, testosterone or dihydrotestosterone (DHT). Men diagnosed with advanced prostate cancer or failure potentially curative therapy are

  20. A quantum mechanical study of TiCl3 alpha, beta and gamma crystal phases: geometry, electronic structure and magnetism.

    PubMed

    Sementa, L; D'Amore, M; Barone, V; Busico, V; Causa', M

    2009-12-21

    The electronic structure of different magnetic states of alpha, beta and gamma modifications of TiCl(3) has been computed employing the density functional theory with periodic boundary conditions and localized Gaussian basis sets. The analysis of the density of the electronic states (DOS) and of the spin density makes it possible to classify these halides as Mott-Hubbard insulators, where the band gap appears a result of large on-site Coulomb interaction. For each crystalline phase, the relative stability of different magnetic states has been analyzed in terms of exchange mechanisms. The electronic population data along with the spin density maps support the assumption of a d(1) Titanium ion in a distorted octahedral crystal field, notwithstanding the not fully ionic character of TiCl(3) modifications. Dispersion forces are particularly important for this material: a classical correction (of the type f(R)/R(6)) has been added to the DFT energies and gradients, providing a good agreement with structural data.

  1. Crystallization and preliminary X-ray diffraction analysis of the interleukin-3 alpha receptor bound to the Fab fragment of antibody CSL362.

    PubMed

    Broughton, Sophie E; Hercus, Timothy R; Nero, Tracy L; Dhagat, Urmi; Owczarek, Catherine M; Hardy, Matthew P; Fabri, Louis J; Scotney, Pierre D; Nash, Andrew D; Wilson, Nicholas J; Lopez, Angel F; Parker, Michael W

    2014-03-01

    Interleukin-3 (IL-3) is a member of the beta common family of cytokines that regulate multiple functions of myeloid cells. The IL-3 receptor-specific alpha subunit (IL3Rα) is overexpressed on stem cells/progenitor cells of patients with acute myeloid leukaemia, where elevated receptor expression correlates clinically with a reduced patient survival rate. The monoclonal antibody (MAb) CSL362 is a humanized MAb derived from the murine MAb 7G3, originally identified for its ability to specifically recognize the human IL-3 receptor and for blocking the signalling of IL-3 in myeloid and endothelial cells. In order to elucidate the molecular mechanism of CSL362 antagonism, a preliminary structure of human IL3Rα in complex with the MAb CSL362 has been determined.

  2. Crystallographic Analysis of Murine Constitutive Androstane Receptor Ligand-Binding Domain Complexed with 5[alpha]-androst-16-en-3[alpha]-ol

    SciTech Connect

    Vincent, J.; Shan, L.; Fan, M.; Brunzelle, J.S.; Forman, B.M.; Fernandez, E.J.

    2010-03-08

    The constitutive androstane receptor (CAR) is a member of the nuclear receptor superfamily. In contrast to classical nuclear receptors, which possess small-molecule ligand-inducible activity, CAR exhibits constitutive transcriptional activity in the apparent absence of ligand. CAR is among the most important transcription factors; it coordinately regulates the expression of microsomal cytochrome P450 genes and other drug-metabolizing enzymes. The murine CAR ligand-binding domain (LBD) was coexpressed with the steroid receptor coactivator protein (SRC-1) receptor-interacting domain (RID) in Escherichia coli. The mCAR LBD subunit was purified away from SRC-1 by affinity, anion-exchange and size-exclusion chromatography, crystallized with androstenol and the structure of the complex determined by molecular replacement.

  3. First-principles study of noncommutative band offsets at [alpha]-Cr2O3/[alpha]-Fe2O3(0001) interfaces

    SciTech Connect

    Jaffe, John E.; Dupuis, Michel; Gutowski, Maciej S.

    2004-05-24

    Using first-principles density functional theory, we have modeled the atomic, electronic and magnetic structure of epitaxial interfaces between alpha-hematite and alpha-chromia (corundum structure) in the hexagonal (0001) basal plane. Our model was a superlattice with a period of about 27.5Å, corresponding to the shortest-period superlattice considered in a recent series of experiments (Chambers et al., Phys. Rev. B 61, 13223 (2000)). Two different epitaxial interface structures were studied: (i) an oxygen plane separating an Fe double layer from a Cr double layer, or (ii) a metal double layer split between Fe and Cr. We found that these two structures are close in total energy but have distinct spin structure and different valence band offsets (chromia above hematite by 0.4 and 0.6 eV for (i) and (ii) respectively), possibly explaining the experimental non-commutative band offset seen in this system (0.3±0.1 eV for hematite grown atop chromia, and 0.7±0.1 eV for the reverse).

  4. Microtubule-associated protein 1 light chain 3 alpha (LC3)-associated phagocytosis is required for the efficient clearance of dead cells

    PubMed Central

    Martinez, Jennifer; Almendinger, Johann; Oberst, Andrew; Ness, Rachel; Dillon, Christopher P.; Fitzgerald, Patrick; Hengartner, Michael O.; Green, Douglas R.

    2011-01-01

    The recognition and clearance of dead cells is a process that must occur efficiently to prevent an autoimmune or inflammatory response. Recently, a process was identified wherein the autophagy machinery is recruited to pathogen-containing phagosomes, termed MAPLC3A (LC3)-associated phagocytosis (LAP), which results in optimal degradation of the phagocytosed cargo. Here, we describe the engagement of LAP upon uptake of apoptotic, necrotic, and RIPK3-dependent necrotic cells by macrophages. This process is dependent on some members of the classical autophagy pathway, including Beclin1, ATG5, and ATG7. In contrast, ULK1, despite being required for autophagy, is dispensable for LAP induced by uptake of microbes or dead cells. LAP is required for efficient degradation of the engulfed corpse, and in the absence of LAP, engulfment of dead cells results in increased production of proinflammatory cytokines and decreased production of anti-inflammatory cytokines. LAP is triggered by engagement of the TIM4 receptor by either phosphatidylserine (PtdSer)-displaying dead cells or PtdSer-containing liposomes. Therefore, the consequence of phagocytosis of dead cells is strongly affected by those components of the autophagy pathway involved in LAP. PMID:21969579

  5. Identification of the NC1 domain of {alpha}3 chain as critical for {alpha}3{alpha}4{alpha}5 type IV collagen network assembly.

    PubMed

    LeBleu, Valerie; Sund, Malin; Sugimoto, Hikaru; Birrane, Gabriel; Kanasaki, Keizo; Finan, Elizabeth; Miller, Caroline A; Gattone, Vincent H; McLaughlin, Heather; Shield, Charles F; Kalluri, Raghu

    2010-12-31

    The network organization of type IV collagen consisting of α3, α4, and α5 chains in the glomerular basement membrane (GBM) is speculated to involve interactions of the triple helical and NC1 domain of individual α-chains, but in vivo evidence is lacking. To specifically address the contribution of the NC1 domain in the GBM collagen network organization, we generated a mouse with specific loss of α3NC1 domain while keeping the triple helical α3 chain intact by connecting it to the human α5NC1 domain. The absence of α3NC1 domain leads to the complete loss of the α4 chain. The α3 collagenous domain is incapable of incorporating the α5 chain, resulting in the impaired organization of the α3α4α5 chain-containing network. Although the α5 chain can assemble with the α1, α2, and α6 chains, such assembly is incapable of functionally replacing the α3α4α5 protomer. This novel approach to explore the assembly type IV collagen in vivo offers novel insights in the specific role of the NC1 domain in the assembly and function of GBM during health and disease.

  6. Direct capture in nuclear astrophysics: The cases of Oxygen-17(p,gamma)Fluorine-18 and Helium-3(alpha,gamma)Beryllium-7

    NASA Astrophysics Data System (ADS)

    Kontos, Antonios

    Direct radiative capture is a non-resonant one-step nuclear reaction mechanism that in principle does not involve the formation of a compound nucleus. In the absence of strong resonance inside the Gamow window, direct capture can be the dominant contribution to astrophysically important reaction rates. The present thesis studies two such reactions, 17O(p,gamma) 18F and 3He(alpha,gamma)7Be. 17O(p,gamma)18F influences hydrogen-burning nucleosynthesis in several stellar sites, such as red giants, asymptotic giant branch (AGB) stars, massive stars and classical novae. In the relevant temperature range for these environments (T9 = 0.01- 0.4), the main contributions to the rate of this reaction are the direct capture process, two low lying narrow resonances (ElabR = 70 and 193 keV) and the low energy tails of two broad resonances (ElabR = 587 and 714 keV). Previous measurements and calculations give contradictory results for the direct capture contribution which in turn increases the uncertainty of the reaction rate. In addition, very few published cross section data exist for the high energy region that might affect the interpretation of the direct capture and the broad resonances contributions in the lower energy range. In this work we present a measurement of the reaction at a wide proton energy range (Elab = 360 - 1625 keV) and at several angles (θ lab = 0°, 45°, 90°, 135°). 3He(alpha,gamma)7Be is important for the neutrino production in the sun's core and the production of 7Li during the big bang nucleosynthesis. Due to the low level density of 7Be, the reaction mechanism is almost entirely non-resonant at the relevant energies. Recent experiments have improved the uncertainty of the reaction but some discrepancies still exist. In the present work, a relatively wide energy window was measured, ECM = 0.300 -- 1.450 MeV, by detecting the prompt gamma-rays from the reaction. The use of a compact helium jet gas target ensured high gamma-ray detection efficiency. The results are compared with literature.

  7. Expression of the human CMP-NeuAc:GM3 alpha2,8-sialyltransferase (GD3 synthase) gene through the NF-kappaB activation in human melanoma SK-MEL-2 cells.

    PubMed

    Kang, Nam-Young; Kim, Cheorl-Ho; Kim, Kyoung-Sook; Ko, Jeong-Heon; Lee, Jai-Heon; Jeong, Yong-Kee; Lee, Young-Choon

    2007-01-01

    To elucidate the mechanism underlying the regulation of human GD3 synthase gene expression in human melanoma SK-MEL-2 cells, we identified the promoter region of the human GD3 synthase gene. The 5'-rapid amplification of cDNA end (5'-RACE) using mRNA prepared from SK-MEL-2 cells revealed the presence of multiple transcription start sites of human GD3 synthase gene. Promoter analyses of the 5'-flanking region of the human GD3 synthase gene using luciferase gene reporter system showed the strong promoter activity in SK-MEL-2 cells. Deletion study revealed that the region as the core promoter from -1146 to -646 (A of the translational start ATG as position +1) was indispensable for endogenous expression of human GD3 synthase gene. This region lacks apparent TATA and CAAT boxes but contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-kappaB. Electrophoretic mobility shift assays using specific competitors, chromatin immunoprecipitation assay and site-directed mutagenesis demonstrated that only NF-kappaB element in this region is required for the promoter activity in SK-MEL-2 cells. These results indicate that NF-kappaB plays an essential role in the transcriptional activity of human GD3 synthase gene essential for GD3 synthesis in SK-MEL-2 cells.

  8. Mutagenesis in the alpha3alpha4 GyrA helix and in the Toprim domain of GyrB refines the contribution of Mycobacterium tuberculosis DNA gyrase to intrinsic resistance to quinolones.

    PubMed

    Matrat, Stéphanie; Aubry, Alexandra; Mayer, Claudine; Jarlier, Vincent; Cambau, Emmanuelle

    2008-08-01

    The replacement of M74 in GyrA, A83 in GyrA, and R447 in GyrB of Mycobacterium tuberculosis gyrase by their Escherichia coli homologs resulted in active enzymes as quinolone susceptible as the E. coli gyrase. This demonstrates that the primary structure of gyrase determines intrinsic quinolone resistance and was supported by a three-dimensional model of N-terminal GyrA.

  9. Platelet activating factor antagonist design. 2. X-ray structure of dimethyl 2,3,4,5-tetrahydro-5 beta-(3,4-methylenedioxyphenyl)-2-oxo-3 beta-(3,4,5-trimethoxybenzoyl)-3 alpha,4 alpha-furandicarboxylate.

    PubMed

    Peterson, J R; Do, H D; Rogers, R D

    1989-07-15

    C25H24O12, Mr = 516.46, triclinic, P-1, a = 8.780 (3), b = 11.298 (4), c = 13.271 (6) A, alpha = 71.77 (4), beta = 70.31 (3), gamma = 72.66 (3) degrees, V = 1189 A3, Z = 2, Dx = 1.44 g cm-3, lambda (Mo K alpha) = 0.71073 A, mu = 0.74 cm-1, F(000) = 540, T = 293 K, final R = 0.046 for 2495 observed [Fo greater than or equal to 5 sigma (Fo)] reflections. The observed structure reveals a trans disposition for the methoxycarbonyl and aryl substituents at positions 4 and 5 of the heterocycle and a cis-3,4-bis(methoxycarbonyl) relationship. There is no crystallographically imposed symmetry. Several intermolecular van der Waals interactions occur in the cell lattice of this compound.

  10. Loss of alpha3/alpha4(IV) collagen from the glomerular basement membrane induces a strain-dependent isoform switch to alpha5alpha6(IV) collagen associated with longer renal survival in Col4a3-/- Alport mice.

    PubMed

    Kang, Jeong Suk; Wang, Xu-Ping; Miner, Jeffrey H; Morello, Roy; Sado, Yoshikazu; Abrahamson, Dale R; Borza, Dorin-Bogdan

    2006-07-01

    Mutations in COL4A3/4/5 genes that affect the normal assembly of the alpha3/4/5(IV) collagen network in the glomerular basement membrane (GBM) cause Alport syndrome. Patients progress to renal failure at variable rates that are determined by the underlying mutation and putative modifier genes. Col4a3(-/-) mice, a model for autosomal recessive Alport syndrome, progress to renal failure significantly slower on the C57BL/6 than on the 129X1/Sv background. Reported here is a novel strain-specific alternative collagen IV isoform switch that is associated with the differential renal survival in Col4a3(-/-) Alport mice. The downregulation or the absence of alpha3/4(IV) collagen chains in the GBM of Lmx1b(-/-) and Col4a3(-/-) mice was found to induce ectopic deposition of alpha5/6(IV) collagen. The GBM deposition of alpha5/6(IV) collagen was abundant in C57BL/6 Col4a3(-/-) mice but almost undetectable in 129X1/Sv Col4a3(-/-) mice. This strain difference was due to overall low expression of alpha6(IV) chain and alpha5/6(IV) protomers in the tissues of 129X1/SvJ mice, a natural Col4a6 knockdown. In (129 x B6)F1 Col4a3(-/-) mice, the amount of alpha5/6(IV) collagen in the GBM was inherited in a mother-to-son manner, suggesting that it is controlled by one or more X-linked loci, possibly Col4a6 itself. Importantly, high levels of ectopic alpha5/6(IV) collagen in the GBM were associated with approximately 46% longer renal survival. These findings suggest that alpha5/6(IV) collagen, the biologic role of which has been hitherto unknown, may partially substitute for alpha3/4/5(IV) collagen. Therapeutically induced GBM deposition of alpha5/6(IV) collagen may provide a novel strategy for delaying renal failure in patients with autosomal recessive Alport syndrome.

  11. Decreased Migration of Langerhans Precursor-Like Cells in Response to Human Keratinocytes Expressing HPV-16 E6/E7 is Related to Reduced Macrophage Inflammatory Protein-3Alpha Production

    DTIC Science & Technology

    2005-01-01

    infection. Introduction. Human papillomaviruses (HPVs) are small DNA viruses which exhibit tropism for cutaneous or mucosal epithelium. Commonly...retinoblastoma protein correlates with the transforming capacity of the E7 oncoproteins of the human papillomaviruses . Proc Natl Acad Sci U S A 89:4442-6

  12. Structural characterization (1->2)-beta-xylose-(1->3)-alpha-arabinose-containing oligosaccharide products of extracted switchgrass (Panicum virgatum, L.) xylan treatment with alpha-arabinofuranosidase and beta-endo-xylanase.

    USDA-ARS?s Scientific Manuscript database

    Switchgrass (Panicum virgatum, L.) is a potential dedicated biomass crop for use in biocatalytic conversion systems to biofuels. Nearly 30% of switchgrass cell wall material is xylan. The complete depolymerization of xylan is desirable both as an additional carbon source for microbial fermentation a...

  13. Comparing prothrombin induced by vitamin K absence-II (PIVKA-II) with the oncofetal proteins glypican-3, Alpha feto protein and carcinoembryonic antigen in diagnosing hepatocellular carcinoma among Egyptian patients.

    PubMed

    Abd El Gawad, Iman A; Mossallam, Ghada I; Radwan, Noha H; Elzawahry, Heba M; Elhifnawy, Niveen M

    2014-06-01

    Hepatocellular carcinoma (HCC) is usually asymptomatic in the early stage and does not show elevated alpha-feto protein (AFP). AFP shows 60-80% sensitivity in diagnosing HCC. Glypican3 (GPC-3) is an oncofetal protein that is only detected in HCC cells but not in benign liver tissues, while Carcinoembryonic antigen (CEA) is expressed in various neoplasms including HCC. Although, it is not specific for HCC. Prothrombin induced by vitamin K absence-II (PIVKA-II) is an abnormal prothrombin protein that is increased in the serum of HCC patients. It has higher sensitivity and specificity compared to AFP. The aim of this study is to compare the clinical utility of PIVKA-II with GPC-3, AFP and CEA in diagnosing HCC. This study included 40 patients with HCC, 10 patients with cirrhosis as a benign control group, and 10 apparently healthy volunteers as normal controls. Serum samples were subjected to routine laboratory investigations, measurement of CEA, AFP using MEIA technique (Axsym), glypican3, and PIVKA-II using ELISA technique in the sera of all patients and controls. All markers showed the highest results in the HCC group. Higher concentrations of PIVKA-II were detected in patients with splenomegaly, and in tumors with size (>3cm). Combination of Glypican-3 and PIVKA-II showed the highest sensitivity, while GPC-3 alone and combination of GPC-3 and AFP showed the highest specificity to differentiate HCC from liver cirrhosis and normal controls. GPC-3, PIVKAII, and combination of both showed the highest sensitivity, while GPC-3 alone showed the highest specificity to differentiate HCC from liver cirrhosis. Glypican-3 is the only oncofetal antigen that showed comparable high diagnostic accuracy as PIVKA-II in diagnosing HCC among Egyptian patients. Copyright © 2014. Production and hosting by Elsevier B.V.

  14. New modified trichothecenes accumulated in solid culture by mutant strains of Fusarium sporotrichioides.

    PubMed Central

    McCormick, S P; Taylor, S L; Plattner, R D; Beremand, M N

    1989-01-01

    Mutant strains of Fusarium sporotrichioides NRRL 3299 deficient in the ability to synthesize T-2 toxin were examined on solid rice medium. Five novel alicyclic trichothecenes were isolated: 11 alpha-hydroxytrichodiene; tricho-9-ene-2 alpha,3 alpha,11 alpha-triol; tricho-9-ene-2 alpha,3 alpha,8 alpha,11 alpha-tetraol; tricho-9-ene-2 alpha,3 alpha,8 beta,11 alpha-tetraol; and tricho-9-ene-2 alpha,3 alpha,11 alpha,16-tetraol. PMID:2802605

  15. Molecular cloning and preliminary expression analysis of banded dogfish (Triakis scyllia) CC chemokine cDNAs by use of suppression subtractive hybridization.

    PubMed

    Inoue, Yuuki; Saito, Tsubasa; Endo, Mariko; Haruta, Chiaki; Nakai, Takeshi; Moritomo, Tadaaki; Nakanishi, Teruyuki

    2005-01-01

    Suppression subtractive hybridization was carried out by using cDNAs of peripheral white blood cells (PWBCs) of banded dogfish (Triakis scyllia) after phorbol 12-myristate 13-acetate (PMA) stimulation. The Trsc-SCYA107, MIP3alpha1 and MIP3alpha2 clones contained an open reading frame encoding 97, 99 and 97 amino acids, respectively. Comparison of the deduced amino acids showed that the banded dogfish MIP3alpha1 and MIP3alpha2 sequences shared 42.3% and 40.0% identity with human SCYA20, respectively, while the Trsc-SCYA107 sequence shared 50.6, 44.2 and 42.0% identity with the catshark (Scyliorhinus canicula) Scca-SCYA107, rainbow trout (Oncorhynchus mykiss) CK4A and CK4B, respectively. The genomic sequences of banded dogfish Trsc-SCYA107, MIP3alpha1 and MIP3alpha2 contain four exons and three introns, and MIP3alpha1 and MIP3alpha2 shared the same intron/exon organization with that of human. The MIP3alpha1 and MIP3alpha2 genes of lipopolysaccharide (LPS)-unstimulated banded dogfish were expressed in gill, kidney and liver, while Trsc-SCYA107 mRNA was detected in various tissues except for brain. However, the constitutive expression of MIP3alpha2 gene was much lower than the Trsc-SCYA107 and MIP3alpha1 genes. RT-PCR analysis of the Trsc-SCYA107 expression in tissues of LPS-stimulated fish showed enhanced expression at 24 h poststimulation in the gill, heart, leydig, spleen and testes, while the expression of MIP3alpha1 and MIP3alpha2 was not influenced by LPS-stimulation in vivo. Furthermore, a relative increase in the expression of the Trsc-SCYA107 and MIP3alpha2 genes in PWBCs was observed at 1-12 h poststimulation with PMA and LPS, with maximal expression observed at 3 h, while MIP3alpha1 expression was observed at 3-12 h poststimulation only with PMA.

  16. Regulation of neurosteroid allopregnanolone biosynthesis in the rat spinal cord by glycine and the alkaloidal analogs strychnine and gelsemine.

    PubMed

    Venard, C; Boujedaini, N; Belon, P; Mensah-Nyagan, A G; Patte-Mensah, C

    2008-04-22

    The neurosteroid allopregnanolone (3alpha,5alpha-THP) is well characterized as a potentially therapeutic molecule which exerts important neurobiological actions including neuroprotective, antidepressant, anxiolytic, anesthetic and analgesic effects. We have recently observed that neurons and glial cells of the rat spinal cord (SC) contain various key steroidogenic enzymes such as 5alpha-reductase and 3alpha-hydroxysteroid oxido-reductase which are crucial for 3alpha,5alpha-THP biosynthesis. Furthermore, we demonstrated that the rat SC actively produces 3alpha,5alpha-THP. As the key factors regulating neurosteroid production by nerve cells are unknown and because glycine is one of the pivotal inhibitory neurotransmitters in the SC, we investigated glycine effects on 3alpha,5alpha-THP biosynthesis in the rat SC. Glycine markedly stimulated [(3)H]-progesterone conversion into [(3)H]3alpha,5alpha-THP by SC slices. The alkaloid strychnine, well-known as a glycine receptor (Gly-R) antagonist, blocked glycine stimulatory effect on 3alpha,5alpha-THP formation. Gelsemine, another alkaloid containing the same functional groups as strychnine, increased 3alpha,5alpha-THP synthesis. The stimulatory effects of glycine and gelsemine on 3alpha,5alpha-THP production were additive when the two drugs were combined. These results demonstrate that glycine and gelsemine, acting via Gly-R, upregulate 3alpha,5alpha-THP biosynthesis in the SC. The data also revealed a structure-activity relationship of the analogs strychnine and gelsemine on neurosteroidogenesis. Possibilities are opened for glycinergic agents and gelsemine utilization to stimulate selectively 3alpha,5alpha-THP biosynthetic pathways in diseases evoked by a decreased neurosteroidogenic activity of nerve cells.

  17. Uptake and metabolism of androgens by bovine spermatozoa.

    PubMed

    Djöseland, O; Høglo, S; Nyberg, K; Hastings, C D; Attramadal, A

    1978-03-01

    Spermatozoa from bovine ejaculates and cauda epiditymidis were incubated with either tritiated 17 beta-hydroxy-5 alpha-androstane-3-one (DHT) or 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol). Examination of the medium incubations demonstrated metabolic conversion of both DHT and 3 alpha-diol when these steriods were incubated with ejaculated sperm. In addition to this interconversion, the following metabolities were identified: 5 alpha-androstane-3 beta, 17 beta-diol, (3 beta-diol), androsterone and 5 alpha-androstane-3, 17-dione (5 alpha-A-dione). Incubations with cauda spermatozoa showed similar metabolic patterns. Androgen binding was exhibited by both sperm types. Examination of the washed cauda sperm pellet, following incubations with 3 alpha-diol showed that the incubated steroid was the most abundantly bound. DHT and 5 alpha-androst-16-en-3 alpha-ol (delta 16-3 alpha-ol1 were also detected. The major part of the radioactivity bound in the sperm pellet was identified as DHT when this steroid was used as the substrate; the remaining radioactivity consisted of 3 alpha-diol and delta 16-3 alpha-ol. Investigations of ejaculated sperm pellets gave similar results apart from the additional identification of 5 alpha-androst-16-en-3 one (delta 16-3-one) and 5 alpha-androst-16-en-3 beta-ol (delta 16-3 beta-ol (delta 16-3 beta-ol).

  18. Mass spectral determination of the configuration of 17[beta]-tetrahydropyranyloxy-19-norandrostan-3[beta]-ol

    NASA Astrophysics Data System (ADS)

    di-Capua, S.; Jang, H. G.; Barasch, D.; Halperin, G.; Deutsch, J.

    1997-11-01

    The stereochemistry of 17[beta]-tetrahydropyranyloxy-19-norandrostan-3[beta]-ol was determined by mass spectral measurements of the extent of water elimination from the molecular ion under electron impact conditions. The [M - H2O][middle dot]+/[M][middle dot]+ ratio of the 3[beta]-OH isomer was found to be three to four times higher than that of the 3[alpha]-OH isomer. The 3[alpha]-OH isomer produced a crystalline 3[alpha]-benzyloxy derivative; its configuration and stereochemistry of the steroidal rings were determined by X-ray crystallography. The 3[alpha]-OH isomer was obtained by reduction of the corresponding carbonyl group with NaBH4. The 3[beta]-OH isomer was synthesized by Walden inversion of the 3[alpha]-OH isomer. The 3[beta] configuration of 17[beta]-tetrahydropyranyloxy-19-norandrostan-3[beta]-ol was also supported by its ability to bind digitonin.

  19. Differential induction of HNF-3 transcription factors during neuronal differentiation.

    PubMed

    Jacob, A; Budhiraja, S; Reichel, R R

    1997-08-01

    We have investigated the regulation of transcription factors HNF-3alpha and HNF-3beta during the retinoic acid-mediated differentiation of mouse P19 cells. Retinoic acid treatment converts P19 stem cells into neurons and astrocytes and we have clearly shown that gene expression of both HNF-3alpha and HNF-3beta is activated during this process. HNF-3alpha transcription was detected 2 h after addition of retinoic acid and took place in the absence of de novo protein synthesis. This suggests that HNF-3alpha is a primary target for retinoic acid action. HNF-3alpha induction displays a biphasic profile and HNF-3alpha mRNA reaches maximal levels at 2 and 6 days postdifferentiation. Additional experiments strongly suggest that the second peak is due to HNF-3alpha induction in postmitotic neurons. P19 stem cells, on the other hand, do not contain any detectable HNF-3alpha mRNA. According to our studies, the retinoic acid-mediated induction of HNF-3alpha occurs at the level of transcriptional initiation and is conferred by distal promoter sequences. In comparison to HNF-3alpha, HNF-3beta induction is a subsequent event and detectable levels of HNF-3beta mRNA materialize approximately 1 day after addition of retinoic acid to P19 stem cells. Time course studies firmly demonstrate that HNF-3beta mRNA peaks at about 2 days postdifferentiation and then declines to virtually unreadable levels. This temporal pattern is consistent with HNF-3beta being a secondary target for retinoic acid. In analogy to HNF-3alpha, HNF-3beta activation also takes place at the level of transcriptional initiation. Recent studies implicate HNF-3alpha and HNF-3beta in early mammalian neurogenesis. The detection of HNF-3alpha/beta activation during P19 cell differentiation provides us with a convenient cell culture system to elucidate the induction mechanism and the precise role of both transcriptional regulators in the formation of neuronal cells.

  20. Alternative splicing switches the divalent cation selectivity of TRPM3 channels.

    PubMed

    Oberwinkler, Johannes; Lis, Annette; Giehl, Klaus M; Flockerzi, Veit; Philipp, Stephan E

    2005-06-10

    TRPM3 is a poorly understood member of the large family of transient receptor potential (TRP) ion channels. Here we describe five novel splice variants of TRPM3, TRPM3alpha1-5. These variants are characterized by a previously unknown amino terminus of 61 residues. The differences between the five variants arise through splice events at three different sites. One of these splice sites might be located in the pore region of the channel as indicated by sequence alignment with other, better-characterized TRP channels. We selected two splice variants, TRPM3alpha1 and TRPM3alpha2, that differ only in this presumed pore region and analyzed their biophysical characteristics after heterologous expression in human embryonic kidney 293 cells. TRPM3alpha1 as well as TRPM3alpha2 induced a novel, outwardly rectifying cationic conductance that was tightly regulated by intracellular Mg(2+). However, these two variants are highly different in their ionic selectivity. Whereas TRPM3alpha1-encoded channels are poorly permeable for divalent cations, TRPM3alpha2-encoded channels are well permeated by Ca(2+) and Mg(2+). Additionally, we found that currents through TRPM3alpha2 are blocked by extracellular monovalent cations, whereas currents through TRPM3alpha1 are not. These differences unambiguously show that TRPM3 proteins constitute a pore-forming channel subunit and localize the position of the ion-conducting pore within the TRPM3 protein. Although the ionic selectivity of ion channels has traditionally been regarded as rather constant for a given channel-encoding gene, our results show that alternative splicing can be a mechanism to produce channels with very different selectivity profiles.

  1. Microbial transformation of the diterpene 7-epi-foliol by Fusarium fujikuroi.

    PubMed

    Fraga, Braulio M; Bressa, Carlo; González, Pedro; Guillermo, Ricardo

    2014-08-01

    The incubation of 3alpha,7alpha,18-trihydroxy-ent-kaur-16-ene (7-epi-foliol) with the fungus Fusarium fujikuroi gave 3alpha,7alpha,18-trihydroxy-ent-kaur-16-en-18-al as the sole product. The biotransformation of other 7alpha- or 7beta-hydroxy derivatives had led to the oxidation of C-19, which is a main step in the biosynthesis of gibberellins and kaurenolides. Now, the presence of the 3alpha-hydroxyl impedes that oxidation, which is directed to the adjacent C-18 hydroxymethyl forming the corresponding aldehyde.

  2. Flavonoids from Aconitum napellus subsp. neomontanum.

    PubMed

    Fico, G; Braca, A; De Tommasi, N; Tomè, F; Morelli, I

    2001-06-01

    Three flavonol glycosides quercetin 7-O-(6-trans-caffeoyl)-beta-glucopyranosyl-(1-->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (1), kaempferol 7-O-(6-trans-caffeoyl)-beta-glucopyranosyl-(1-->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (2), and kaempferol 7-O-(6-trans-p-coumaroyl)-beta-glucopyranosyl-(1-->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (3), together with the known beta-3,4-dihydroxyphenethyl beta-glucopyranoside, were isolated from the flowers of Aconitum napellus subsp. neomontanum. Their structures were elucidated by spectroscopic methods, including 2D NMR spectral techniques.

  3. Morphometric analysis of prostates in castrated dogs after treatment with androstanediol, estradiol, and cyproterone acetate.

    PubMed

    Tunn, U W; Schüring, B; Senge, T; Neumann, F; Schweikert, H U; Rohr, H P

    1981-01-01

    We studied the prostates of 22 beagle dogs by light morphometric analysis under defined hormonal influences. Prostatic weight increased with 3 alpha-androstanediol (3 alpha-diol) alone and in combination with 17 beta-estradiol (E2). Two different prostatic hyperplasia models were established. 3 alpha-Diol brings about diffuse glandular prostatic hyperplasia with an absolute increase of glandular parenchyma and, together with E2, provokes stromal hyperplasia with squamous metaplasia of the epithelium and secondary cyst formation. In both models, the antiandrogen cyproterone acetate causes a significant reduction of the absolute and relative volumes of the glandular compartment with antiandrogenic effects on the stroma.

  4. Synthesis and GABA(A) receptor activity of 2,19-sulfamoyl analogues of allopregnanolone.

    PubMed

    Durán, Fernando J; Edelsztein, Valeria C; Ghini, Alberto A; Rey, Mariana; Coirini, Héctor; Dauban, Philippe; Dodd, Robert H; Burton, Gerardo

    2009-09-15

    The synthesis of new analogues of allopregnanolone with a bridged sulfamidate ring over the beta-face of ring A has been achieved from easily available precursors, using an intramolecular aziridination strategy. The methodology also allows the synthesis of 3alpha-substituted analogues such as the 3alpha-fluoro derivative. GABA(A) receptor activity of the synthetic analogues was evaluated by assaying their effect on the binding of [(3)H]flunitrazepam and [(3)H]muscimol. The 3alpha-hydroxy-2,19-sulfamoyl analogue and its N-benzyl derivative were more active than allopregnanolone for stimulating binding of [(3)H]flunitrazepam. For the binding of [(3)H]muscimol, both synthetic analogues and allopregnanolone stimulated binding to a similar extent, with the N-benzyl derivative exhibiting a higher EC(50). The 3alpha-fluoro derivative was inactive in both assays.

  5. Bile salts of the green turtle Chelonia mydas (L.)

    PubMed Central

    Haslewood, G A; Ikawa, S; Tökés, L; Wong, D

    1978-01-01

    1. Bile salts of the green turtle Chelonia mydas (L.) were analysed as completely as possible. 2. They consist of taurine conjugates of 3 alpha, 7 alpha, 12 alpha, 22 xi-tetrahydroxy-5 beta-cholestan-26-oic acid (tetrahydroxysterocholanic acid) and 3 alpha 12 alpha, 22 xi-trihydroxy-5 beta-cholestan-26-oic acid, with minor amounts of 3 alpha, 7 alpha, 12 alpha-trihydroxy-5beta-cholan-24-oic acid (cholic acid), 3alpha, 12 alpha-dihydroxy-5beta-cholan-24-oic acid (deoxycholic acid) and possibly other bile acids. 3. Cholic acid and deoxycholic acid represent the first known examples of bile acids common to chelonians and other animal forms: they may indicate independent evolution in chelonians to C24 bile acids. 4. The discovery of a 7-deoxy C27 bile acid is the first evidence that C27 bile acids or their conjugates have an enterohepatic circulation. PMID:656052

  6. The role of neurosteroids and non-genomic effects of progestins and androgens in mediating sexual receptivity of rodents.

    PubMed

    Frye, C A

    2001-11-01

    Progestins and androgens modulate sexual receptivity in rodents, in part through mechanisms independent of traditional intracellular steroid receptors. Progesterone (PROG) in the ventromedial hypothalamus (VMH) and ventral tegmental (VTA) facilitates lordosis but has different actions in these brain areas. Primarily using lordosis in rodents as an in vivo experimental model, we have examined the effects that progestins exert in the midbrain and hypothalamus. Localization and blocker studies indicate that PROG's actions in the VMH require intracellular progestin receptors (PRs) but in the VTA they do not. Progestins that have rapid, membrane effects, and/or are devoid of affinity for PRs, facilitate lordosis when applied to the VTA. Manipulation of GABA and/or GABA(A)/benzodiazepine receptor complexes (GBRs) in the VTA alters lordosis, which suggests that progestins may interact with GBRs to facilitate receptivity by enhancing the function of GABAergic neurons. Interfering with PROG's metabolism to, or the biosynthesis of, 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-TH PROG or allopregnanolone), the most effective endogenous GBR agonist, in the VTA attenuates female sexual behavior in rodents. Stimulation of mitochondrial benzodiazepine receptors (MBRs), which enhances neurosteroid production, by infusions of an MBR agonist to the VTA enhances lordosis. 3alpha,5alpha-TH PROG is increased in the midbrain of mated>proestrous>diestrous rodents. These data suggest that in the VTA, PROG may facilitate lordosis following metabolism to and/or biosynthesis of 3alpha,5alpha-TH PROG, which may have subsequent actions at GBRs and/or MBRs to acutely modulate female sexual behavior in rodents. The 3alpha-hydroxysteroid oxidoreduced metabolite of dihydrotestosterone (DHT), 5alpha-androstane-3alpha,17beta-diol (3alpha-androstanediol), is important for termination of sexual receptivity in rodents and has these effects in the absence of functional intracellular androgens

  7. Effects of neurosteroids on epileptiform activity induced by picrotoxin and 4-aminopyridine in the rat hippocampal slice.

    PubMed

    Salazar, Patricia; Tapia, Ricardo; Rogawski, Michael A

    2003-01-01

    The neurosteroids allopregnanolone (5alpha-pregnan-3alpha-ol-20-one; 5alpha,3alpha-P) and its 5beta-epimer pregnanolone (5beta,3alpha-P), and pregnenolone sulfate (PS) were examined for effects on spontaneous epileptiform discharges induced by 100 microM picrotoxin (PTX) and 55 microM 4-aminopyridine (4-AP) in the CA3 region of the rat hippocampal slice. At a concentration of 10 microM, 5alpha,3alpha-P partially reduced PTX-induced bursting and at 30 and 90 microM completely suppressed bursting. In contrast, 100 microM 5beta,3alpha-P failed to alter the discharge frequency. 5alpha,3alpha-P depressed 4-AP-induced bursting with similar potency as in the PTX model; 100 microM 5beta,3alpha-P was also partially effective. In the 4-AP model, 5alpha,3alpha-P inhibited both the more frequent predominantly positive-going potentials as well as the less frequent negative-going potentials that may be generated by synchronous GABAergic interneuron firing. PS enhanced the PTX bursting frequency and, in the 4-AP model, increased the frequency of negative potentials but did not alter the frequency of positive potentials. By itself, PS did not induce bursting. The effects of the steroids in the in vitro seizure models largely correspond with their activities on GABA(A) receptors; suppression of discharges may occur as a result of direct activation of these receptors rather than modulation of GABA-mediated synaptic responses. PTX and 4-AP-induced bursting in the hippocampal slice are useful models for directly assessing neurosteroid effects on seizure susceptibility under conditions that eliminate the factor of brain bioavailability.

  8. Complete 1H and 13C NMR assignments of six saponins from Sapindus trifoliatus.

    PubMed

    Grover, Rajesh K; Roy, Abhijeet D; Roy, Raja; Joshi, S K; Srivastava, Vandita; Arora, Sudershan K

    2005-12-01

    Complete 1H and 13C spectral assignments are reported for six saponins from the pericarp of Sapindus trifoliatus (Hindi name: Reetha) collected from Madhya Pradesh and Maharashtra, India, using only 1D and 2D NMR methods. The structures of the compounds were elucidated as hederagenin 3-O-(3-O-acetyl-beta-D-xylopyranosyl)-(1-3)-alpha-L-rhamnopyranosyl-(1-2)-alpha-L-ara-binopyranoside, hederagenin 3-O-(4-O-acetyl-beta-D-xylop-yranosyl)-(1-3)-alpha-L-rhamnopyranosyl-(1-2)-alpha-L-arabinop-yranoside, hederagenin 3-O-(3,4-O-diacetyl-beta-D-xylopy-ranosyl)-(1-3)-alpha-L-rhamnopyranosyl-(1-2)-alpha-L-arabinopy-ranoside, hederagenin 3-O-(3,4-O-diacetyl-alpha-L-arabinop-yranosyl)-(1-3)-alpha-L-rhamnopyranosyl-(1-2)-alpha-L-arabinop-yranoside, hederagenin 3-O-(beta-D-xylopyranosyl)-(1-3)-alpha-L-rhamnopyranosyl-(1-2)-alpha-L-arabinopyranoside and he-deragenin 3-O-(alpha-L-arabinopyranosyl)-(1-3)-alpha-L-rhamno-pyranosyl-(1-2)-alpha-L-arabinopyranoside. It is concluded that saponins of this complexity approach the limit of structural complexity, which can be solved by NMR alone, precisely and quickly.

  9. Molluscicidal saponins from Sapindus mukorossi, inhibitory agents of golden apple snails, Pomacea canaliculata.

    PubMed

    Huang, Hui-Chi; Liao, Sin-Chung; Chang, Fang-Rong; Kuo, Yao-Haur; Wu, Yang-Chang

    2003-08-13

    Extracts of soapnut, Sapindus mukorossi Gaertn. (Sapindaceae) showed molluscicidal effects against the golden apple snail, Pomacea canaliculata Lamarck. (Ampullariidae) with LC(50) values of 85, 22, and 17 ppm after treating 24, 48, and 72 h, respectively. Bioassay-directed fractionation of S. mukorossi resulted in the isolation of one new hederagenin-based acetylated saponin, hederagenin 3-O-(2,4-O-di-acetyl-alpha-l-arabinopyranoside)-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (1), along with six known hederagenin saponins, hederagenin 3-O-(3,4-O-di-acetyl-alpha-l-arabinopyranoside)-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (2), hederagenin 3-O-(3-O-acetyl-beta-d-xylopyranosyl)-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (3), hederagenin 3-O-(4-O-acetyl-beta-d-xylopyranosyl)-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (4), hederagenin 3-O-(3,4-O-di-acetyl-beta-d-xylopyranosyl)-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (5), hederagenin 3-O-beta-d-xylopyranosyl-(1-->3)-alpha-l-rhamnopyranosyl-(1-->2)-alpha-l-arabinopyranoside (6), and hederagenin 3-O-alpha-l-arabinopyranoside (7). The bioassay data revealed that 1-7 were molluscicidal, causing 70-100% mortality at 10 ppm against the golden apple snail.

  10. 21beta-Hydroxy-oleanane-type triterpenes from Hippocratea excelsa.

    PubMed

    Cáceres-Castillo, David; Mena-Rejón, Gonzalo J; Cedillo-Rivera, Roberto; Quijano, Leovigildo

    2008-02-01

    Stem bark of Hippocratea excelsa afforded six pentacyclic triterpenes, five oleanane and one ursane types. They were identified as 11beta,21beta-dihydroxy-olean-12-ene-3-one (2), 3alpha,11alpha,21beta-trihydroxy-olean-12-ene (3), 3alpha,21beta-dihydroxy-11alpha-methoxy-olean-12-ene (4), 3alpha,21beta-dihydroxy-olean-9(11),12-diene (5), 3alpha,21beta-dihydroxy-olean-12-ene (6) and 3alpha,21beta-dihydroxy-11alpha-methoxy-urs-12-ene, isolated as its diacetate derivative (7), as well as 3alpha,21beta-dihydroxy-olean-12-ene (1) previously isolated from the root bark. The known alpha- and beta-amyrin, oleanoic and ursolic acids, trans-polyisoprene, and the ubiquitous beta-sitosterol were also isolated. Structures were elucidated on the basis of spectroscopic analyses, including homo- and heteronuclear correlation NMR experiments (COSY, ROESY, HSQC and HMBC) and comparison with literature data. The antigiardial activity of compounds 2-5 was not significant.

  11. Metabolism of the contraceptive steroid desogestrel by human liver in vitro.

    PubMed

    Madden, S; Back, D J; Orme, M L

    1990-02-01

    The metabolism of the progestogen oral contraceptive desogestrel (Dg) has been studied in vitro using human liver microsomes. Metabolites have been separated using radiometric high performance liquid chromatography and identified by co-chromatography with authentic standards and by mass spectrometry. All the livers examined (n = 6) were able to form 3-keto desogestrel as the main identifiable metabolite and also the presumed intermediates 3 alpha-hydroxydesogestrel (3 alpha-OHDg) and 3 beta-hydroxydesogestrel (3 beta-OHDg). In addition, a large polar heterogenous peak was evident on the radiochromatograms which did not co-chromatograph with any known metabolites of desogestrel. Inter-individual variability in metabolite formation was seen. A number of drugs were examined for their propensity to inhibit desogestrel metabolism. Primaquine was the most potent tested having an IC50 value (inhibitory concentration reducing overall metabolite production by 50%) of 30 microM. Cimetidine, trilostane and levonorgestrel failed to inhibit at 250 microM. With 3 alpha-OHDg as substrate, 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSDH) activity was 1.0 +/- 0.3 nmol min-1 mg-1 protein which was five times greater than the activity of the 3 beta-HSDH towards 3 beta-OHDg. Miconazole was the most potent inhibitor tested having IC50 values of 14 and 95 microM for 3 alpha- and 3 beta-HSDH respectively. Surprisingly, trilostane was without inhibitory effect on either enzyme, which contrasts with other data involving 3 beta-HSDH in steroidogenic tissue. Our observations with trilostane may reflect tissue differences in the enzyme and/or differences in endogenous vs exogenous steroids (i.e. in the conversion of 3 beta-OHDg to 3-ketodesogestrel there is no requirement for isomerization). Kinetic parameters of 3 alpha-HSDH were also determined.

  12. Chemotaxonomy of the pantropical genus Merremia (Convolvulaceae) based on the distribution of tropane alkaloids.

    PubMed

    Jenett-Siems, Kristina; Weigl, Robert; Böhm, Anke; Mann, Petra; Tofern-Reblin, Britta; Ott, Sonja C; Ghomian, Azar; Kaloga, Maki; Siems, Karsten; Witte, Ludger; Hilker, Monika; Müller, Frank; Eich, Eckart

    2005-06-01

    The occurrence and distribution of tropane and biogenetically related pyrrolidine alkaloids in 18 Merremia species of paleo-, neo-, and pantropical occurrence have been studied. The extensive GC-MS study included members of almost all sections of the genus and has been carried out with epigeal vegetative parts as well as with roots. It comprises altogether 74 tropanes and 13 pyrrolidines including nicotine. Along with datumetine known already from a solanaceous species, the study led to the isolation (from M. dissecta and M. guerichii, respectively) and structure elucidation (spectral data) of four novel 3alpha-acyloxytropanes, merresectines A-D: 3alpha-(4-methoxybenzoyloxy)nortropane (A), 3alpha-kurameroyloxytropane (B), 3alpha-nervogenoyloxytropane (C), 3alpha-[4-(beta-D-glucopyranosyloxy)-3-methoxy-5-(3-methyl-2-butenyl)benzoyloxy]tropane (beta-d-glucoside of D). Moreover, the novel 3alpha,6beta-di-(4-methoxybenzoyloxy)tropane (merredissine) has been isolated from M. dissecta and structurally elucidated. In addition the structures of datumetine and merresectine A could be confirmed by synthesis. Spectral data for two known 3alpha-acyloxytropanes (merresectine E beta-D-glucoside, 4'-dihydroconsabatine) and one known 3beta-acyloxytropane (concneorine) are documented for the first time. The structures of three further merresectines (F-H) have been determined by mass spectrometry. Furthermore, the linkage (2',3- and 2',4-, respectively) of two position isomer N-methylpyrrolidinylhygrines was proven by synthesis. The results of the study contribute to the solution of infrageneric taxonomic problems. Whereas all species yield pyrrolidine alkaloids without suitably differentiating results the diverging occurrence of tropane alkaloids leads to three groups of sections: (1) taxa free of tropanes, (2) taxa with simple tropanes, and (3) taxa with merresectines in addition to simple tropanes.

  13. Effect of neuroactive steroids on [3H]flumazenil binding to the GABAA receptor complex in vitro.

    PubMed

    Bertz, R J; Reynolds, I J; Kroboth, P D

    1995-09-01

    Modulation of benzodiazepine receptor ligand binding to the GABAA receptor complex by the neuroactive steroids 3 alpha-hydroxy-dihydroprogesterone (3 alpha-OH-DHP) and 3 alpha-hydroxycorticosterone (3 alpha- THDOC) was assessed in an in vitro binding assay with the benzodiazepine antagonist [3H]flumazenil using rat cortical membranes. Neuroactive steroids, pentobarbital, GABA and bicuculline did not significantly affect flumazenil binding. However, the addition of neuroactive steroids significantly decreased the Ki of benzodiazepine agonists, including alprazolam, diazepam and clonazepam, indicating an increase in agonist affinity. Only the addition of 3 beta-OH-DHP, an inactive stereoisomer had no effect on the Ki of these agonists. The binding of the benzodiazepine inverse agonist FG 7142 was not significantly affected by these steroids, but the addition of GABA significantly increased the Ki of FG 7142 indicating a decrease in inverse agonist affinity. High concentrations of GABA or bicuculline were able to occlude the 3 alpha-THDOC mediated decrease in alprasolam Ki, indicating a GABA dependent mechanism of binding enhancement. An advantage of using [3H]flumazenil is that neither the Ki nor the Bmax change in the presence of allosteric site modulators, permitting the simple and direct assessment of alterations in benzodiazepine ligand affinity for the GABAA receptor complex by neuroactive steroids.

  14. Virilizing para-adrenocortical adenoma associated with idiopathic-acquired generalized anhidrosis in an adolescent girl.

    PubMed

    Gumus, Pinar; Luquette, Mark; Haymon, Marie Louise; Valerie, Evans; Morales, Jaime; Vargas, Alfonso

    2011-01-01

    Adrenocortical tumors are rare in childhood and adolescence. Virilization, alone or in combination with signs of overproduction of other adrenal hormones, is the most common clinical presentation. Here we report an unusual case of an African-American female adolescent presenting with idiopathic acquired generalized anhidrosis, dysregulation of body temperature, absence of adult body odor and dry skin in the face of a virilizing para-adrenocortical adenoma. Virilization signs regressed soon after removal of the tumor, but normalization of the 3alpha-androstenediol glucuronide (3alpha-AG) took longer compared to other measurable androgens; accompanied by anhidrosis. The association of remitting anhidrosis with normalized levels of 3alpha-AG suggests it might be a possible mechanism for anhidrosis. High 3alpha-AG levels might implicate the increased peripheral conversion of weak pro-androgens with different biochemical structure. We recommend obtaining 3alpha-AG beside other androgens in virilized patients with atypical dermatological symptoms in the face of hyperandrogenism.

  15. Radicals in 5-methylcytosine induced by ionizing radiation. Electron magnetic resonance for structural and mechanistic analyses.

    PubMed

    Krivokapić, André; Ohman, Kjell Tage; Munthe, Morten; Nelson, William H; Hole, Eli O; Sagstuen, Einar

    2010-05-01

    Single crystals of 5-methylcytosine hemihydrate and 5-methylcytosine hydrochloride were X-irradiated and studied at 10 K and at higher temperatures using X- and K-band EPR, ENDOR and EIE spectroscopy. In the hemihydrate crystals, four radicals were identified at 10 K, one of them being the recently reported N1-deprotonated one-electron oxidation product (Krivokapić et al., J. Phys. Chem. A 113, 9633-9640, 2009). The other radicals were the 3alphaH radical and the C6 and C5 H-addition radicals (the 5-yl and 6-yl radicals, respectively). After irradiation at 295 K, only the 3alphaH and the 5-yl radicals were observed. In the hydrochloride crystals, at least seven different radicals were present after irradiation at 10 K. These were the N1-deprotonated one-electron oxidation product, the 3alphaH radical, three different one-electron reduction products, and the 5- and 6-yl radicals. DFT calculations were used to assist in assigning the observed couplings. The 3alphaH and 5-yl radicals were dominant after thermal annealing to room temperature. In neither crystal system did the N1-deprotonated oxidation product transform into the 3alphaH radical upon warming. The radical yield was significantly greater after irradiation at 300 K compared to that after irradiation at 10 K followed by warming to 300 K and was also considerably greater in the hydrochloride crystals than in the hemihydrate crystals.

  16. Triple-alpha reaction rate studied with the Faddeev three-body formalism

    SciTech Connect

    Ishikawa, Souichi

    2012-11-12

    The triple-alpha (3{alpha}) reaction, {sup 4}He+{sup 4}He+{sup 4}He{yields}{sup 12}C+{gamma}, which plays a significant role in the stellar evolution scenarios, is studied in terms of a three-alpha (3-{alpha}) model. The reaction rate of the process is calculated via an inverse process, 3-{alpha} photodisintegration of a {sup 12}C nucleus. Both of 3-{alpha} bound and-continuum states are calculated by a Faddeev method with accommodating the long range Coulomb interaction. With being adjusted to the empirical E2-strength for {sup 12}C(0{sub 2}{sup +}){yields}{sup 12}C(2{sub 1}{sup +}) transition, results of the 3{alpha} reaction rate <{alpha}{alpha}{alpha}> at higher temperature (T > 10{sup 8} K), where the reaction proceeds mainly through the {sup 8}Be and {sup 12}C(0{sub 2}{sup +}) resonant states, almost agree with those of the Nuclear Astrophysics Compilation of Reaction Rates (NACRE). On the other hand, calculated values of <{alpha}{alpha}{alpha}> are about 10{sup 3} times larger than the NACRE rate at a low temperature (T= 10{sup 7} K), which means our results are remarkably smaller than recent CDCC results.

  17. Chronic intermittent ethanol (CIE) administration in rats decreases levels of neurosteroids in hippocampus, accompanied by altered behavioral responses to neurosteroids and memory function.

    PubMed

    Cagetti, Elisabetta; Pinna, Graziano; Guidotti, Alessandro; Baicy, Kate; Olsen, Richard W

    2004-03-01

    The administration of ethanol on a chronic intermittent regimen (CIE) involving multiple withdrawal episodes is a model for ethanol dependence. After CIE, rats exhibited reduced seizure threshold, increased anxiety, tolerance to GABAergic sedative-hypnotic drugs, and changes in GABA(A) receptor function and subunit composition in hippocampus. Previous studies have shown that acute and chronic ethanol may induce changes in the levels of the neurosteroid 3alpha-hydroxysteroid-5alpha-pregnan-20-one (3alpha, 5alpha-THP) in the brain. Therefore, the current study analyses the correlation between chronic intermittent ethanol effects on the level of 3alpha, 5alpha-THP in hippocampus of CIE rats and the behavioral changes in sensitivity to neurosteroids. After CIE, the levels for 3alpha, 5alpha-THP were significantly reduced in hippocampus of rats. The mRNA levels for the enzymes 5alpha-reductase and 3alpha-HSD in hippocampus were also reduced. In vivo, (in contrast to a tolerance to the hypnotic effect of steroids), CIE rats showed increased sensitivity to the anticonvulsant and to the anxiolytic effect of the steroid alphaxalone. Perhaps, this is a response to lowered levels of endogenous neuroactive steroids. CIE rats also showed impairment of hippocampus-dependent memory function. These results suggest that changes in neurosteroids level and in vivo sensitivity to these compounds are involved in the development of ethanol dependence in the CIE model.

  18. New steroidal lactones and homomonoterpenic glucoside from fruits of Malva sylvestris L.

    PubMed

    Mustafa, Akhlaq; Ali, Mohammed

    2011-01-01

    Phytochemical investigation of the ethanolic extract of defatted fruits of Malva sylvestris Linn. (Malvaceae) led to the isolation of six new steroidal lactones and a homomonoterpenic glucoside along with beta-sitosterol-3-beta-D-glucopyranoside. The structures of new phytoconstituents have been elucidated as cholest-5-en-3a-ol-18(21)-olide (sylvestrosterol A), cholest-9(11)-en-3alpha-ol-18(21)-olide (sylvestrosterol B), cholest-4,6,22-trien-3alpha-ol-18(21)-olide (sylvestrosterol C), 2-methyl-6-methylene-n-decan-2-olyl- 3beta-D-glucopyranoside (malvanoyl glucoside), cholest-7-en-18(21)-olide-3alpha-olyl-3beta-D-glucopyranoside (sylvestrogenin A), cholest-9(11)-en-18(21)-olide-3alpha-olyl-3beta-D-glucopyranoside (sylvestrogenin B) and cholest-5-en-8(21)-olide-3alpha-olyl-3beta-D-glucopyranoside (sylvestrogenin C).The structures of all these phytoconstituents have been established on the basis of spectral data analysis and chemical reactions.

  19. A theoretical analysis of the coordination modes of CuII with penicillins: activation of the beta-lactam C-N bond.

    PubMed

    Campomanes, Pablo; Menéndez, M Isabel; López, Ramón; Sordo, Tomás L

    2005-02-01

    The interaction of CuII with 6-formylamino-3alpha-carboxypenam and 6-acetylamino-3alpha-carboxypenam was investigated by means of DFT calculations with the UB3LYP functional. Nine different modes of complexation between CuII and 6-formylamino-3alpha-carboxypenam were located. When two water molecules directly bonded to CuII are included in the calculations on 6-acetylamino-3alpha-carboxypenam as penicillin model, only six CuII(H2O)2-6-acetylamino-3alpha-carboxypenam complexes (1S-6S) are found. In solution the four most stable complexes obtained from our calculations, 6S, 1S, 2S, and 3S, exhibit CuII in square-planar coordination with at least one bond to the carboxylate group, in agreement with experimental evidence. Complexes 6S, 1S, and 3S were previously suggested by available experimental evidence. In three of the most stable complexes (6S, 2S, and 3S) the beta-lactam C-N bond is remarkably activated and displays C-N bond lengths similar to those found in some tetrahedral intermediates located for the hydrolysis of 2-azetidinones. This suggests that these kinds of complexes belong to the reaction coordinate for the degradation of beta-lactam antibiotics in the presence of CuII.

  20. Trinervitene diterpenes from soldiers of two Nasutitermes species from French Guyana.

    PubMed

    Laurent, P; Daloze, D; Pasteels, J M; Braekman, J C

    2005-04-01

    Methanolic extracts of soldiers of Nasutitermes guayanae and N. surinamensis have been shown to contain complex mixtures of diterpenes and monoterpenes. Eighteen diterpenes have been isolated and identified; twelve of them are previously known nasute termite diterpenes, while six are new trinervitene diterpenes. 2alpha,9beta-Dihydroxy-3beta,8beta-oxido-1(15)-trinervitene has been isolated from N. guayanae, while 3alpha,14alpha-diacetoxy-2beta-hydroxy-1(15),8(19),9-trinervitatriene, 14alpha-acetoxy-2beta,3alpha-dihydroxy-1(15),8(19),9-trinervitatriene, 2beta,3alpha-diacetoxy-11beta,14alpha-dihydroxy-1(15),8(19)-trinervitadiene, 9alpha,14alpha-diacetoxy-2beta,3alpha-dihydroxy-1(15),8(19)-trinervitadiene, and 2beta,9alpha,14alpha-triacetoxy-3alpha-hydroxy-1(15),8(19)-trinervitadiene have been isolated from N. surinamensis. Their structures were determined on the basis of their spectroscopic properties.

  1. Acetylated triterpene saponins from the Thai medicinal plant, Sapindus emarginatus.

    PubMed

    Kanchanapoom, T; Kasai, R; Yamasaki, K

    2001-09-01

    From the pericarps of Sapindus emarginatus (Sapindaceae), three new acetylated triterpene saponins were isolated together with hederagenin and five known triterpene saponins, as well as one known sweet acyclic sesquiterpene glycoside, mukurozioside IIb. The structures of new compounds were elucidated as hederagenin 3-O-(2-O-acetyl-beta-D-xylopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside, 23-O-acetyl-hederagenin 3-O-(4-O-acetyl-beta-D-xylopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside and oleanolic acid 3-O-(4-O-acetyl-beta-D-xylopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside by chemical and spectroscopic data.

  2. Structure of a glucosyl phosphate-containing O-polysaccharide of Proteus vulgaris O42.

    PubMed

    Perepelov, Andrei V; Bartodziejska, Beata; Shashkov, Alexander S; Wykrota, Marianna; Knirel, Yuriy A; Rozalski, Antoni

    2007-12-28

    An O-polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Proteus vulgaris O42 and studied by sugar and methylation analyses along with 1H, 13C and 31P NMR spectroscopy. The following structure of the polysaccharide having a linear pentasaccharide phosphate repeating unit was established: -->3)-alpha-L-FucpNAc4Ac-(1-->4)-alpha-D-Glcp-1-P-(O-->4)-alpha-D-GlcpNAc-(1-->3)-alpha-L-FucpNAc4Ac-(1-->3))-alpha-D-GlcpNAc6Ac-(1--> where the degree of O-acetylation is approximately 80% on GlcNAc and approximately 40% on each of the FucNAc residues. A weak serological cross-reaction of anti-P. vulgaris O42 serum with the lipopolysaccharide of P. vulgaris O39 was observed and accounted for by the sharing of a disaccharide fragment of the O-polysaccharides.

  3. Factors affecting the pheromone composition of voided boar saliva.

    PubMed

    Booth, W D

    1987-11-01

    The pheromone binding protein 'pheromaxein' which binds the pheromonal 16-androstene steroids in the saliva of the male pig (boar), was degraded and lost its binding activity in saliva incubated in air for 72 h at 21 degrees C and 37 degrees C. However, pheromaxein and its binding activity were retained in saliva incubated for 168 h at 4 degrees C. When the 3H-labelled pheromones 5 alpha-androst-16-en-3 alpha-ol (3 alpha-androstenol), 5 alpha-androst-16-en-3-one (5 alpha-androstenone) and 5 alpha-androst-16-en-3 beta-ol (3 beta-androstenol) were incubated with boar saliva for 168 h at 21 degrees C, 3 alpha-androstenol was primarily converted to 5 alpha-androstenone and 5 alpha-androstenone to 3 beta-androstenol; 3 beta-androstenol was unchanged. Evidence was obtained for microorganisms being responsible for these steroid transformations.

  4. Differential inhibition of aflatoxin B1 oxidation by gestodene action on human liver microsomes.

    PubMed

    Kim, B R; Oh, H S; Kim, D H

    1997-11-01

    Human cytochrome P450 (P450) 3A is known to be involved in the formation of both aflatoxin B1-exo-8,9-epoxide (exo-epoxidation) and aflatoxin Q1 (3 alpha-hydroxylation). Gestodene, a known inactivator of P450 3A4, inhibited the formation of AFB1 metabolites in a variety of ways depending on the incubation condition. Preincubation of gestodene with human liver microsomes prior to the addition of AFB1 inhibited both exo-epoxidation and 3 alpha-hydroxylation whereas simultaneous incubation of gestodene with AFB1 only inhibited 3 alpha-hydroxylation. These results suggest that two independent substrate binding sites exist in P450 3A4, and AFB1 binds to both of the binding sites. Gestodene selectively binds to one of the binding sites leading to the formation of AFQ1, whereas it does not affect the formation of exo-epoxide via the other binding site.

  5. Triterpenoids from the leaves of Diospyros kaki (persimmon) and their inhibitory effects on protein tyrosine phosphatase 1B.

    PubMed

    Thuong, Phuong Thien; Lee, Chul Ho; Dao, Trong Tuan; Nguyen, Phi Hung; Kim, Wan Gi; Lee, Sang Jun; Oh, Won Keun

    2008-10-01

    Phytochemical study on a methanol-soluble extract of the leaves of persimmon (Diospyros kaki) resulted in the isolation of two new ursane-type triterpenoids, 3alpha,19alpha-dihydroxyurs-12,20(30)-dien-24,28-dioic acid (1) and 3alpha,19alpha-dihydroxyurs-12-en-24,28-dioic acid (2), together with 12 known ursane- and oleanane-type triterpenoids (3-14). Triterpenoids with a 3beta-hydroxy group were found to inhibit protein tyrosine phosphatase 1B (PTP1B) activity, with IC50 values ranging from 3.1+/-0.2 to 18.8+/-1.3 microM, whereas those with a 3alpha-hydroxy moiety were not active.

  6. Enzymic synthesis of oligosaccharides terminating in the tumor-associated sialyl-Lewis-a determinant.

    PubMed

    Palcic, M M; Venot, A P; Ratcliffe, R M; Hindsgaul, O

    1989-07-01

    The isomeric sialyl-Lea-terminating pentasaccharide derivatives, alpha-Neup5Ac-(2----3)-beta-D-Galp-(1----3)-[alpha-L-Fucp-(1 ----4)]-beta- D-GlcpNAc-(1----3)-beta-D-Galp-O(CH2)8COOMe and alpha-Neup5Ac-(2----3)-beta-D-Galp-(1----3)-[alpha-L-Fucp-(1 ----4)]- beta-D-GlcpNAc-(1----6)-beta-D-Galp-O(CH2)8COOMe, have been prepared by the action in sequence of a porcine submaxillary (2----3)-alpha-sialyltransferase and a human-milk (1----3/4)-alpha-fucosyltransferase on the chemically synthesized trisaccharides beta-D-Galp-(1----3)-beta-D-GlcpNAc-(1----3)- and -(1----6)-beta-D-Galp- O(CH2)8COOMe, respectively.

  7. Effect of progesterone, testosterone and their 5 alpha-reduced metabolites on GFAP gene expression in type 1 astrocytes.

    PubMed

    Melcangi, R C; Riva, M A; Fumagalli, F; Magnaghi, V; Racagni, G; Martini, L

    1996-03-04

    Astrocytes possess steroid receptors as well as several enzymes typical of steroid target cells, such as 5 alpha-reductase, which converts testosterone (T) and progesterone (P) into their respective 5 alpha-reduced metabolites, and the 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD). Because of this, it was deemed of interest to analyze whether the original hormones P and T, and their 5 alpha-reduced metabolites dihydrotestosterone (DHT), 5 alpha-androstan-3 alpha, 17 beta-diol (3 alpha-diol), dihydroprogesterone (DHP) and 5 alpha-pregnan-3 alpha-ol-20-one (THP), might exert some effects on the expression of the most typical astrocytic marker, i.e. the glial fibrillary acidic protein (GFAP). Cultures of rat type 1 astrocytes were exposed to the various steroids for 2, 6, and 24 h, and the variations of GFAP mRNA were measured by Northern blot analysis. A significant elevation of GFAP mRNA levels was observed after exposure to either P or DHP; the effect of DHP appeared more promptly (at 2 h) than that of P (at 6 h). This result suggests that the effect of P might be linked to its conversion into DHP; this hypothesis has been confirmed by showing that the addition of finasteride (a specific blocker of the 5 alpha-reductase) is able to completely abolish the effect of P. After exposure to DHP or THP, a decrease of GFAP gene expression was observed at later intervals (24 h). In the case of androgens, T and 3 alpha-diol did not change GFAP expression at any time of exposure, while DHT produced a significant decrease of GFAP mRNA only after 24 h of exposure. Taken together, the data indicate that the 5 alpha-reduced metabolites of P and T may modulate the expression of GFAP in type 1 rat astrocytes.

  8. Cross metathesis for the synthesis of novel C-sialosides.

    PubMed

    Meinke, Sebastian; Thiem, Joachim

    2008-07-21

    Cross metathesis of both anomers of C-allyl sialoside (3alpha/3beta) with styrene catalyzed by the second generation Grubbs or Hoveyda-Grubbs catalysts gave the corresponding aryl derivatives (4alpha/4beta) in virtually quantitative yields. The products were hydrogenated to model compounds 5alpha/5beta. Similarly, reaction of the alpha-anomer 3alpha with galactose derivative 8 gave the olefin-linked disaccharide mimetic 9. Following hydrogenation and deprotection, the ethylene-bridged Neu5Acalpha(2-->6)Gal analogue 11 could be obtained.

  9. [Biological activity tests of chemical constituents from two Brazilian Labiatae plants].

    PubMed

    Isobe, Takahiko; Doe, Matsumi; Morimoto, Yoshiki; Nagata, Kumiko; Masuoka, Noriyoshi; Ohsaki, Ayumi

    2007-02-01

    We studied the bioactivities of constituents from two tropical medicinal plants, Cunila spicata and Hyptis fasciculata. These plants found in Brazil belong to the Labiatae family. Four known compounds obtained from these herbs were identified as 3alpha, 24-dihydoxylurs-12-en-28-oic acid, betulinic acid, aurantiamide acetate, and aurantiamide benzoate by spectroscopic means. 3alpha, 24-Dihydoxylurs-12-en-28-oic acid has potent inhibitory activity against Streptococcus salivarius, Streptococcus pneumoniae, Streptococcus pyogenes, and Porphyromomas gingivalis. Aurantiamide benzoate exhibited moderate inhibitory activity against xanthine oxidase. It was clarified that herbs Cunila spicata and Hyptis fasciculata are effective against bronchitis and gout.

  10. Insulin receptor substrate-2 phosphorylation is necessary for protein kinase C zeta activation by insulin in L6hIR cells.

    PubMed

    Oriente, F; Formisano, P; Miele, C; Fiory, F; Maitan, M A; Vigliotta, G; Trencia, A; Santopietro, S; Caruso, M; Van Obberghen, E; Beguinot, F

    2001-10-05

    We have investigated glycogen synthase (GS) activation in L6hIR cells expressing a peptide corresponding to the kinase regulatory loop binding domain of insulin receptor substrate-2 (IRS-2) (KRLB). In several clones of these cells (B2, F4), insulin-dependent binding of the KRLB to insulin receptors was accompanied by a block of IRS-2, but not IRS-1, phosphorylation, and insulin receptor binding. GS activation by insulin was also inhibited by >70% in these cells (p < 0.001). The impairment of GS activation was paralleled by a similarly sized inhibition of glycogen synthase kinase 3 alpha (GSK3 alpha) and GSK3 beta inactivation by insulin with no change in protein phosphatase 1 activity. PDK1 (a phosphatidylinositol trisphosphate-dependent kinase) and Akt/protein kinase B (PKB) activation by insulin showed no difference in B2, F4, and in control L6hIR cells. At variance, insulin did not activate PKC zeta in B2 and F4 cells. In L6hIR, inhibition of PKC zeta activity by either a PKC zeta antisense or a dominant negative mutant also reduced by 75% insulin inactivation of GSK3 alpha and -beta (p < 0.001) and insulin stimulation of GS (p < 0.002), similar to Akt/PKB inhibition. In L6hIR, insulin induced protein kinase C zeta (PKC zeta) co-precipitation with GSK3 alpha and beta. PKC zeta also phosphorylated GSK3 alpha and -beta. Alone, these events did not significantly affect GSK3 alpha and -beta activities. Inhibition of PKC zeta activity, however, reduced Akt/PKB phosphorylation of the key serine sites on GSK3 alpha and -beta by >80% (p < 0.001) and prevented full GSK3 inactivation by insulin. Thus, IRS-2, not IRS-1, signals insulin activation of GS in the L6hIR skeletal muscle cells. In these cells, insulin inhibition of GSK3 alpha and -beta requires dual phosphorylation by both Akt/PKB and PKC zeta.

  11. Hasty effect on the metabolism of glycyrrhizin by Eubacterium sp. GLH with Ruminococcus sp. PO1-3 and Clostridium innocuum ES24-06 of human intestinal bacteria.

    PubMed

    Akao, T

    2000-01-01

    Eubacterium sp. GLH with Ruminococcus sp. PO1-3 and Clostridium innocuum ES24-06 possessing enzymes involved in the metabolism of glycyrrhizin (GL) was cultured in GAM medium with and without 1.0 mM GL or 1.0 mM glycyrrhetic acid (GA). GL (1.0 mM) enhanced 3alpha-hydroxyglycyrrhetinate (3alpha-hydroxyGA) dehydrogenase activity, GA (1.0 mm) suppressed 3alpha-hydroxyGA dehydrogenase activity, GL beta-D-glucuronidase activity and the mixed bacterial growth, and GL and GA showed almost no change in a lower level of 3beta-hydroxysteroid dehydrogenase (3beta-HSD) activity during 5 d of culture. GL (1.0 mM) and GA (1.0 mM) were metabolized to a small amount of GA and a negligible amount of 3-oxo-glycyrrhetic acid (3-oxo-GA) and 3alpha-hydroxyGA, and to a negligible amount of 3-oxo-GA, respectively, by these mixed bacteria. These amounts coincided with those of metabolites produced from 1.0 mM GL and 1.0 mM GA added to these mixed bacteria after 24 h culture. Whole bacteria and sonicated bacteria derived from the collection of these mixed bacteria reached a maximal stage and metabolized GL to a relatively large amount of GA and 3-oxo-GA, and a negligible amount of 3alpha-hydroxyGA and GA to a small amount of 3-oxo-GA and 3alpha-hydroxyGA within 180 min. GL beta-D-glucuronidase with 3beta-HSD and 3alpha-hydroxyGA dehydrogenase partially purified from each bacterium was converted GL to 3alpha-hydroxyGA, producing metabolites of about 60% after 10 min of incubation. These mixed bacteria possessed high enzyme activities could produce the metabolites of GL in under one hour under conditions.

  12. Fluoxetine-elicited changes in brain neurosteroid content measured by negative ion mass fragmentography.

    PubMed Central

    Uzunov, D P; Cooper, T B; Costa, E; Guidotti, A

    1996-01-01

    Fluoxetine administered intraperitoneally to sham-operated or adrenalectomized/castrated (ADX/CX) male rats dose-dependently (2.9-58 mumol/kg i.p.) increased the brain content of the neurosteroid 3 alpha-hydroxy-5 alpha-pregnan-20-one (allopregnanolone, 3 alpha, 5 alpha-TH PROG). The increase of brain 3 alpha, 5 alpha-TH PROG content elicited by 58 mumol/kg fluoxetine lasted more than 2 hr and the range of its extent was comparable in sham-operated (approximately 3-10 pmol/g) and ADX/CX rats (2-9 pmol/g) and was associated with a decrease (from 2.8 to 1.1 pmol/g) in the 5 alpha-pregnan-3,20-dione (5 alpha-dihydroprogesterone, 5 alpha-DH PROG) content. The pregnenolone, progesterone, and dehydroepiandrosterone content failed to change in rats receiving fluoxetine. The extent of 3 alpha, 5 alpha-TH PROG accumulation elicited by fluoxetine treatment differed in various brain regions, with the highest increase occurring in the olfactory bulb. Importantly, fluoxetine failed to change the 3 alpha, 5 alpha-TH PROG levels in plasma, which in ADX/CX rats were at least two orders of magnitude lower than in the brain. Two other serotonin re-uptake inhibitors, paroxetine and imipramine, in doses equipotent to those of fluoxetine in inhibiting brain serotonin uptake, were either significantly less potent than fluoxetine (paroxetine) or failed to increase (imipramine) 3 alpha, 5 alpha-TH PROG brain content. The addition of 10 microM of 5 alpha-DH PROG to brain slices of ADX/CX rats preincubated with fluoxetine (10 microM, 15 min) elicited an accumulation of 3 alpha, 5 alpha-TH PROG greater than in slices preincubated with vehicle. A fluoxetine stimulation of brain 3 alpha, 5 alpha-TH PROG biosynthesis might be operative in the anxiolytic and antidysphoric actions of this drug. Images Fig. 4 Fig. 5 Fig. 6 PMID:8901628

  13. Coumarins and a naphthyl labdanoate diarabinoside from the fruits of Peucedanum grande C. B. Clarke.

    PubMed

    Aslam, Mohammad; Ali, Mohammed; Dayal, Rameshwar; Javed, Kalim

    2012-01-01

    Phytochemical investigations of the methanolic extract of the fruits of Peucedanum grande C. B. Clarke (Apiaceae) led to the identification of three coumarins and a naphthyl labdanoate diarabinoside characterized as 5-hydroxy-6-isopranyl coumarin (1), 5,6-furanocoumarin (2), 7-methoxy-5,6-furanocoumarin (3), and labdanyl-3alpha-ol-18-(3'"-methoxy-2"'-naphthyl-oate)-3alpha-L-arabinofuranosyl-(2'-->1")-alpha-L-arabinofuranoside (4). The structures of these compounds were identified on the basis of spectral data analysis and chemical reactions. The methanolic extract and 4 showed nephroprotective activity against gentamicin-induced nephrotoxicity in Wistar rats.

  14. Synthesis of oligosaccharides corresponding to structures found in capsular polysaccharides of Cryptococcus neoformans--II.

    PubMed

    Garegg, P J; Olsson, L; Oscarson, S

    1996-11-01

    Formula 1 depicts a generalized structure of the capsular polysaccharides of four serotypes of the opportunistic microorganism Cryptococcus neoformans, which appears as one of the major infections in the late stages of development of AIDS. Syntheses are now described of two tetrasaccharides with corresponding structures. These are methyl O-alpha-D-mannopyranosyl-(1-->3)-[O-beta-D-xylopyranosyl-(1-->2)] -O-alpha-D-mannopyranosyl-(1-->3)-alpha-D-mannopyranoside and methyl O-alpha-D-mannopyranosyl-(1-->3)-[O-beta-D-glucopyranosyluronic acid-(1-->2)]-O-alpha-D-mannopyranosyl-(1-->3)-alpha-D-mannopyranoside.

  15. Release of free and conjugated forms of the putative pheromonal steroid 11-oxo-etiocholanolone by reproductively mature male round goby (Neogobius melanostomus Pallas, 1814).

    PubMed

    Katare, Yogesh Kumar; Scott, Alexander P; Laframboise, Alyson J; Li, Weiming; Alyasha'e, Zena; Caputo, Christopher B; Loeb, Stephen J; Zielinski, Barbara

    2011-02-01

    Previous studies of the round goby (Neogobius melanostomus Pallas, 1814), an invasive fish species in the Laurentian Great Lakes of North America, have shown that this species has the ability to both synthesize and smell steroids that have a 5 beta-reduced and 3 alpha-hydroxyl (5 beta,3 alpha) configuration. An enzyme-linked immunoassay (EIA) for 3 alpha-hydroxy-5 beta-androstane-11,17-dione (11-O-ETIO) has been used to show a substantial rise in the rate of release of immunoreactive compounds into the water when males are injected with salmon gonadotropin releasing hormone analogue. Similar increases were noted for 11-ketotestosterone and 17,20 beta-dihydroxypregn-4-en-3-one. Partitioning of the extracts between diethyl ether and water showed the presence of both free and conjugated immunoreactive 11-O-ETIO. Only conjugated immunoreactivity was found in urine (implying that free steroid is released via the gills). The identity of the conjugates was probed by using HPLC, EIA, and mass spectrometry and removal of sulfate and glucosiduronate groups. Immunoreactivity in the conjugated fraction was found to be due mainly to 3 alpha,17beta-dihydroxy-5 beta-androstan-11-one 17-sulfate. However, the evidence was also strong for the presence in water extracts of substantial amounts of 3 alpha-hydroxy-5 beta-androstane-11,17-dione 3-glucosiduronate (which could be detected only by EIA after removal of the glucosiduronate group with beta-glucuronidase). There were also small amounts of 3 alpha-hydroxy-5 beta-androstane-11,17-dione 3-sulfate and 3 alpha,17beta-dihydroxy-5 beta-androstan-11-one 17-glucosiduronate. These studies give some idea of the types, amounts, and ratios of 11-O-ETIO derivatives that are released by reproductive N. melanostomus and will aid further research into the putative pheromonal roles of 5 beta,3 alpha-reduced androgens in this species.

  16. Indonesian medicinal plants. IX. Chemical structures of gongganosides A, B, and C, three new quinovic acid glycosides from the bark of Bhesa paniculata (Celastraceae).

    PubMed

    Ohashi, K; Kojima, H; Tanikawa, T; Okumura, Y; Kawazoe, K; Tatara, N; Shibuya, H; Kitagawa, I

    1994-08-01

    Three new quinovic acid glycosides, named gongganosides A (1), B (2), and C (3), were isolated from the bark of Bhesa paniculata (Celastraceae), an Indonesian medicinal plant collected in Sumatra Island. The chemical structures have been elucidated on the basis of chemical and physicochemical evidence as quinovic acid 3-O-beta-D-xylopyranosyl(1-->3)-alpha-L- rhamnopyranoside for 1,28-O-beta-D-glucopyranosylquinovic acid 3-O-alpha-L-rhamnopyranoside for 2, and 28-O-beta-D-glucopyranosyl-quinovic acid 3-O-beta-D-xylopyranosyl(1-->3)-alpha-L-rhamnopyranoside for 3.

  17. Two new sesquiterpene lactones from Montanoa tomentosa ssp. microcephala.

    PubMed

    Braca, A; Cioffi, G; Morelli, I; Venturella, F; Pizza, C; De Tommasi, N

    2001-11-01

    Two new sesquiterpene lactones: 8alpha-(4'-acetoxymethacryloyloxy)-3alpha,9beta-dihydroxy-1(10)E,4Z,11(13)-germacratrien-12,6alpha-olide (1) and 8alpha-(2'E)-(2'-acetoxymethyl-2'-butenoyloxy)-3alpha,9beta-dihydroxy-1(10)E,4Z,11(13)-germacratrien-12,6alphaolide (2), together with the known zoapatanolide A were isolated from the aerial parts of Montanoa tomentosa Cerv. in La Llave et Lex ssp. microcephala (Sch. Bip. In K. Koch) V.A. Funk (Asteraceae). The structures of all compounds were established on the basis of 1D, 2D NMR, and EIMS analysis.

  18. Simplified quantitative determination of total fecal bile acids.

    PubMed

    de Wael, J; Raaymakers, C E; Endeman, H J

    1977-09-01

    To determine total fecal bile acids, these are extracted with diethyl ether after boiling with a solution of potassium hydroxide in ethanediol. After evaporating the ether and dissolving the residue in methanol, the bile acids are directly determined with 3 alpha-hydroxysteroid dehydrogenase. Values for 9 normals are given.

  19. Vasodilating effect of norethisterone and its 5 alpha metabolites: a novel nongenomic action.

    PubMed

    Perusquía, Mercedes; Villalón, Carlos M; Navarrete, Erika; García, Gustavo A; Pérez-Palacios, Gregorio; Lemus, Ana E

    2003-08-15

    Estrogens are generally administered in hormone replacement therapy in combination with synthetic progestins. Studies of cardiovascular risk factors in postmenopausal women have shown a variety of responses according to the molecular structure of the progestin used in hormone replacement therapy schemes. The present study sets out to determine the vasoactive effects of norethisterone and its 5alpha-dihydro (5alpha-norethisterone) and -tetrahydro (3alpha,5alpha-norethisterone and 3beta,5alpha-norethisterone) metabolites in isolated precontracted rat thoracic aorta. The addition of norethisterone and 3alpha,5alpha-norethisterone in rat aorta exhibited a potent, concentration-response inhibition of noradrenaline-induced contraction, while 5alpha- and 3beta,5alpha-norethisterone had very little, if any, vasorelaxing effect. Relaxation to norethisterone and 3alpha,5alpha-norethisterone had very rapid time-courses and it was neither affected by the absence of endothelium nor by the inhibitor of nitric oxide synthase, Nomega-nitro-L-arginine methyl ester (L-NAME). The addition of specific anti-androgen, anti-progestin and anti-estrogen compounds and protein synthesis inhibitors did not preclude the vasorelaxing effect of norethisterone and its 3alpha,5alpha-reduced metabolite. The results strongly suggest that these effects are not mediated by nuclear sex steroid hormone receptors. The overall data document a novel nongenomic endothelium-independent vasorelaxing action of a 19-nor synthetic progestin and one of its A-ring-reduced derivatives.

  20. In Vitro Fermentation of Oligosaccharides from Raffinose and Alternansucrase by Human Intestinal Bacteria

    USDA-ARS?s Scientific Manuscript database

    The alternansucrases are enzymes involved in the synthesis of oligosaccharides with alternating glycosidic alpha-(1'3), alpha-(1'6) bonds by reactions where there is a donor of glucosyl residues (sucrose) and an acceptor. There are many carbohydrates that have been used as acceptors, including raff...

  1. 40 CFR 180.950 - Tolerance exemptions for minimal risk active and inert ingredients.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... agricultural or manufacturing practices. Chemical CAS No. Acetic acid, sodium salt 127-09-3 Alpha-cyclodextrin..., sodium salt 532-32-1 Beta-cyclodextrin 7585-39-9 Carbonic acid, monopotassium salt 298-14-6 Carbonic acid, monosodium salt (sodium bicarbonate) 144-55-8 Carnauba wax 8015-86-9 Carob gum (locust bean gum) 9000-40-2...

  2. 40 CFR 180.950 - Tolerance exemptions for minimal risk active and inert ingredients.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... agricultural or manufacturing practices. Chemical CAS No. Acetic acid, sodium salt 127-09-3 Alpha-cyclodextrin..., sodium salt 532-32-1 Beta-cyclodextrin 7585-39-9 Carbonic acid, monopotassium salt 298-14-6 Carbonic acid, monosodium salt (sodium bicarbonate) 144-55-8 Carnauba wax 8015-86-9 Carob gum (locust bean gum) 9000-40-2...

  3. 40 CFR 180.950 - Tolerance exemptions for minimal risk active and inert ingredients.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... agricultural or manufacturing practices. Chemical CAS No. Acetic acid, sodium salt 127-09-3 Alpha-cyclodextrin..., sodium salt 532-32-1 Beta-cyclodextrin 7585-39-9 Carbonic acid, monopotassium salt 298-14-6 Carbonic acid, monosodium salt (sodium bicarbonate) 144-55-8 Carnauba wax 8015-86-9 Carob gum (locust bean gum) 9000-40-2...

  4. 40 CFR 180.950 - Tolerance exemptions for minimal risk active and inert ingredients.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... agricultural or manufacturing practices. Chemical CAS No. Acetic acid, sodium salt 127-09-3 Alpha-cyclodextrin..., sodium salt 532-32-1 Beta-cyclodextrin 7585-39-9 Carbonic acid, monopotassium salt 298-14-6 Carbonic acid, monosodium salt (sodium bicarbonate) 144-55-8 Carnauba wax 8015-86-9 Carob gum (locust bean gum) 9000-40-2...

  5. Triterpenoid saponins from the fruits and galls of Sapindus mukorossi.

    PubMed

    Huang, Hui-Chi; Wu, Ming-Der; Tsai, Wei-Jern; Liao, Sin-Chung; Liaw, Chia-Ching; Hsu, Li-Chuan; Wu, Yang-Chang; Kuo, Yao-Haur

    2008-05-01

    Six saponins, sapinmusaponin K (1) [hederagenin-3-O-(3-O-acetyl-alpha-L-arabinopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], sapinmusaponin L (2) [hederagenin-3-O-(4-O-acetyl-alpha-L-arabinopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabino-pyranoside], sapinmusaponin M (3) [hederagenin-3-O-(2,3-O-diacetyl-beta-D-xylopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], sapinmusaponin N (4) [hederagenin-3-O-(2,4-O-diacetyl-beta-D-xylopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside], sapinmusaponin O (5) [3,7,20(S)-trihydroxydammar-24-ene-3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranoside], and sapinmusaponin P (6) [3,7,20(R)-trihydroxydammar-24-ene-3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-d-glucopyranoside], along with seven known saponins (7-13), were isolated from fruits and the galls of Sapindus mukorossi. Their structures were elucidated by 1D and 2D NMR spectroscopic techniques and acid hydrolysis. Biological evaluation indicated that saponins 1-4 and 7-13 showed moderate cytotoxicity against several human tumor cell lines.

  6. On the Extraction of Components and the Applicability of the Factor Model.

    ERIC Educational Resources Information Center

    Dziuban, Charles D.; Harris, Chester W.

    A reanalysis of Shaycroft's matrix of intercorrelations of 10 test variables plus 4 random variables is discussed. Three different procedures were used in the reanalysis: (1) Image Component Analysis, (2) Uniqueness Rescaling Factor Analysis, and (3) Alpha Factor Analysis. The results of these analyses are presented in tables. It is concluded from…

  7. Transport of resistance-inducing sterols in phloem sap of barley.

    PubMed

    Lehrer, A T; Dugassa-Gobena, D; Vidal, S; Seifert, K

    2000-01-01

    After root application of [7alpha-3H]-7beta-hydroxysitosterol and [3alpha,6beta-3H2]-6alpha-hydroxylathosterol these sterols could be detected in the leaves and phloem sap feeding aphids. These results imply that the phloem sap is a sterol transport system in barley plants.

  8. Comparative analysis of fibrillar and basement membrane collagen expression in embryos of the sea urchin, Strongylocentrotus purpuratus.

    PubMed

    Suzuki, H R; Reiter, R S; D'Alessio, M; Di Liberto, M; Ramirez, F; Exposito, J Y; Gambino, R; Solursh, M

    1997-06-01

    The time of appearance and location of three distinct collagen gene transcripts termed 1 alpha, 2 alpha, and 3 alpha, were monitored in the developing S. purpuratus embryo by in situ hybridization. The 1 alpha and 2 alpha transcripts of fibrillar collagens were detected simultaneously in the primary (PMC) and secondary (SMC) mesenchyme cells of the late gastrula stage and subsequently expressed in the spicules and gut associated cells of the pluteus stage. The 3 alpha transcripts of the basement membrane collagen appeared earlier than 1 alpha and 2 alpha, and were first detected in the presumptive PMC at the vegetal plate of the late blastula stage. The PMC exhibited high expression of 3 alpha at the mesenchyme blastula stage, but during gastrulation the level of expression was reduced differentially among the PMC. In the late gastrula and pluteus stages, both PMC and SMC expressed 3 alpha mRNA, and thus at these stages all three collagen genes displayed an identical expression pattern by coincidence. This study thus provides the first survey of onset and localization of multiple collagen transcripts in a single sea urchin species.

  9. Interactions of five D-mannose-specific lectins with a series of synthetic branched trisaccharides.

    PubMed

    Kaku, H; Goldstein, I J; Oscarson, S

    1991-06-25

    The interaction of a series of synthetic, branched trisaccharides with five D-mannose-specific lectins was studied by precipitation-inhibition assay. The branched methyl alpha-D-mannotrioside, alpha-D-Manp-(1----3)-[alpha-D-Manp-(1----6)]-alpha-D-Man pOMe, the best inhibitor of the Con A-Dextran interaction, was 42 times more potent than alpha-D-ManpOMe, and 3-6 times more potent than the two trisaccharides substituted with D-glucosyl groups, and 8-15 times those with D-galactosyl groups. Surprisingly, methyl O-alpha-D-mannopyranosyl-(1----3)-alpha-D-mannopyranoside was bound to Con A 8-fold more avidly than methyl alpha-D-mannopyranoside. However, the related pea lectin (PSA) was singularly different from Con A in its carbohydrate-binding activity, showing no significantly enhanced binding to any of the sugars examined. The trisacchrides containing terminal, nonreducing, (1----3)-linked alpha-D-mannopyranosyl groups, i.e., alpha-D-Manp-(1----3)-[alpha-D-Glep-(1----6)]alpha-D-Manp OMe, alpha-D-Manp-(1----3)]-alpha-D-Galp-(1----6)]-alpha-D-ManpOMe++ +, and alpha-D-Manp-(1----3)-[alpha-D-Manp-(1----6)]-alpha-D-Man pOMe, were the best inhibitors of the snowdrop lectin (GNA)-D-mannan precipitation system. On the other hand, all branched trisaccharides exhibited very similar inhibitory potencies toward the daffodil lectin (NPA)-D-mannan interaction, whereas alpha-D-Manp-(1----3)-[alpha-D-Galp-(1----6)]-alpha-D-ManpOMe++ + and alpha-D-Manp-(1----3)-[alpha-D-Manp-(1----6)]-alpha-D-Man pOMe were somewhat better inhibitors than the other branched trisaccharides of the amaryllis lectin (HHA)-D-mannan precipitation reaction. (ABSTRACT TRUNCATED AT 250 WORDS)

  10. Androgen metabolism and regulation of rat ventral prostate growth and acid phosphatase during sexual maturation.

    PubMed

    Orlowski, J; Bird, C E; Clark, A F

    1988-01-01

    Androgen metabolism and the regulation of rat ventral prostate cell proliferation and secretory function were examined during sexual maturation. Changes in acid phosphatase (AP) characteristics were measured as a marker of androgen-dependent prostatic secretory function. In immature (21-day-old) rats, total AP activity per cell was low (14.2 +/- 1.3 mol p-nitrophenol phosphate hydrolysed/h per mg DNA); it increased threefold as the weight, protein and DNA contents of the prostate increased to adult (65-day) levels. This corresponded with significant (P less than 0.001) increases in the staining intensities of three of the four bands of secretory AP on isoelectric focusing gels. The extent of inhibition of AP by tartrate decreased at the same time. Secretory AP is known to be relatively tartrate-resistant. The changes in AP activity occurred after prostatic 5 alpha-dihydrotestosterone (5 alpha-DHT) levels increased from 4.6 +/- 0.7 pmol/mg DNA (21 days) to reach a peak of 17.6 +/- 2.3 pmol/mg DNA at 58 days. Prostatic 5 alpha-DHT concentrations were always higher than testosterone levels. Prostatic 5 alpha-androstane-3 alpha,17 beta-diol (3 alpha-Adiol) levels were lower than 5 alpha-DHT levels except on day 58 when levels peaked dramatically at 26.2 +/- 5.5 pmol/mg DNA. Changes in prostatic 5 alpha-DHT and 3 alpha-Adiol levels corresponded with changes in 5 alpha-reductase and 3 alpha-hydroxysteroid oxidoreductase (3 alpha-HSOR) activities. The oxidative reaction of 3 alpha-HSOR was approximately fourfold higher than the reductive reaction, indicating a preference for the formation of 5 alpha-DHT. The plasma levels of testosterone, 5 alpha-DHT and 3 alpha-Adiol cannot account for their respective prostatic levels, indicating the importance of the steroid-metabolizing enzymes in regulating intracellular androgen levels. Changes in the AP characteristics could be correlated with the androgen status of the prostate.

  11. THE IMPACT OF HELIUM-BURNING REACTION RATES ON MASSIVE STAR EVOLUTION AND NUCLEOSYNTHESIS

    SciTech Connect

    West, Christopher; Heger, Alexander; Austin, Sam M. E-mail: alexander.heger@monash.edu

    2013-05-20

    We study the sensitivity of presupernova evolution and supernova nucleosynthesis yields of massive stars to variations of the helium-burning reaction rates within the range of their uncertainties. The current solar abundances from Lodders are used for the initial stellar composition. We compute a grid of 12 initial stellar masses and 176 models per stellar mass to explore the effects of independently varying the {sup 12}C({alpha}, {gamma}){sup 16}O and 3{alpha} reaction rates, denoted R{sub {alpha},12} and R{sub 3{alpha}}, respectively. The production factors of both the intermediate-mass elements (A = 16-40) and the s-only isotopes along the weak s-process path ({sup 70}Ge, {sup 76}Se, {sup 80}Kr, {sup 82}Kr, {sup 86}Sr, and {sup 87}Sr) were found to be in reasonable agreement with predictions for variations of R{sub 3{alpha}} and R{sub {alpha},12} of {+-}25%; the s-only isotopes, however, tend to favor higher values of R{sub 3{alpha}} than the intermediate-mass isotopes. The experimental uncertainty (one standard deviation) in R{sub 3{alpha}}(R{sub {alpha},12}) is approximately {+-}10%({+-}25%). The results show that a more accurate measurement of one of these rates would decrease the uncertainty in the other as inferred from the present calculations. We also observe sharp changes in production factors and standard deviations for small changes in the reaction rates, due to differences in the convection structure of the star. The compactness parameter was used to assess which models would likely explode as successful supernovae, and hence contribute explosive nucleosynthesis yields. We also provide the approximate remnant masses for each model and the carbon mass fractions at the end of core-helium burning as a key parameter for later evolution stages.

  12. The N-end rule pathway catalyzes a major fraction of the protein degradation in skeletal muscle

    NASA Technical Reports Server (NTRS)

    Solomon, V.; Lecker, S. H.; Goldberg, A. L.

    1998-01-01

    In skeletal muscle, overall protein degradation involves the ubiquitin-proteasome system. One property of a protein that leads to rapid ubiquitin-dependent degradation is the presence of a basic, acidic, or bulky hydrophobic residue at its N terminus. However, in normal cells, substrates for this N-end rule pathway, which involves ubiquitin carrier protein (E2) E214k and ubiquitin-protein ligase (E3) E3alpha, have remained unclear. Surprisingly, in soluble extracts of rabbit muscle, we found that competitive inhibitors of E3alpha markedly inhibited the 125I-ubiquitin conjugation and ATP-dependent degradation of endogenous proteins. These inhibitors appear to selectively inhibit E3alpha, since they blocked degradation of 125I-lysozyme, a model N-end rule substrate, but did not affect the degradation of proteins whose ubiquitination involved other E3s. The addition of several E2s or E3alpha to the muscle extracts stimulated overall proteolysis and ubiquitination, but only the stimulation by E3alpha or E214k was sensitive to these inhibitors. A similar general inhibition of ubiquitin conjugation to endogenous proteins was observed with a dominant negative inhibitor of E214k. Certain substrates of the N-end rule pathway are degraded after their tRNA-dependent arginylation. We found that adding RNase A to muscle extracts reduced the ATP-dependent proteolysis of endogenous proteins, and supplying tRNA partially restored this process. Finally, although in muscle extracts the N-end rule pathway catalyzes most ubiquitin conjugation, it makes only a minor contribution to overall protein ubiquitination in HeLa cell extracts.

  13. Ubiquitin conjugation by the N-end rule pathway and mRNAs for its components increase in muscles of diabetic rats

    NASA Technical Reports Server (NTRS)

    Lecker, S. H.; Solomon, V.; Price, S. R.; Kwon, Y. T.; Mitch, W. E.; Goldberg, A. L.

    1999-01-01

    Insulin deficiency (e.g., in acute diabetes or fasting) is associated with enhanced protein breakdown in skeletal muscle leading to muscle wasting. Because recent studies have suggested that this increased proteolysis is due to activation of the ubiquitin-proteasome (Ub-proteasome) pathway, we investigated whether diabetes is associated with an increased rate of Ub conjugation to muscle protein. Muscle extracts from streptozotocin-induced insulin-deficient rats contained greater amounts of Ub-conjugated proteins than extracts from control animals and also 40-50% greater rates of conjugation of (125)I-Ub to endogenous muscle proteins. This enhanced Ub-conjugation occurred mainly through the N-end rule pathway that involves E2(14k) and E3alpha. A specific substrate of this pathway, alpha-lactalbumin, was ubiquitinated faster in the diabetic extracts, and a dominant negative form of E2(14k) inhibited this increase in ubiquitination rates. Both E2(14k) and E3alpha were shown to be rate-limiting for Ub conjugation because adding small amounts of either to extracts stimulated Ub conjugation. Furthermore, mRNA for E2(14k) and E3alpha (but not E1) were elevated 2-fold in muscles from diabetic rats, although no significant increase in E2(14k) and E3alpha content could be detected by immunoblot or activity assays. The simplest interpretation of these results is that small increases in both E2(14k) and E3alpha in muscles of insulin-deficient animals together accelerate Ub conjugation and protein degradation by the N-end rule pathway, the same pathway activated in cancer cachexia, sepsis, and hyperthyroidism.

  14. Ubiquitin conjugation by the N-end rule pathway and mRNAs for its components increase in muscles of diabetic rats

    NASA Technical Reports Server (NTRS)

    Lecker, S. H.; Solomon, V.; Price, S. R.; Kwon, Y. T.; Mitch, W. E.; Goldberg, A. L.

    1999-01-01

    Insulin deficiency (e.g., in acute diabetes or fasting) is associated with enhanced protein breakdown in skeletal muscle leading to muscle wasting. Because recent studies have suggested that this increased proteolysis is due to activation of the ubiquitin-proteasome (Ub-proteasome) pathway, we investigated whether diabetes is associated with an increased rate of Ub conjugation to muscle protein. Muscle extracts from streptozotocin-induced insulin-deficient rats contained greater amounts of Ub-conjugated proteins than extracts from control animals and also 40-50% greater rates of conjugation of (125)I-Ub to endogenous muscle proteins. This enhanced Ub-conjugation occurred mainly through the N-end rule pathway that involves E2(14k) and E3alpha. A specific substrate of this pathway, alpha-lactalbumin, was ubiquitinated faster in the diabetic extracts, and a dominant negative form of E2(14k) inhibited this increase in ubiquitination rates. Both E2(14k) and E3alpha were shown to be rate-limiting for Ub conjugation because adding small amounts of either to extracts stimulated Ub conjugation. Furthermore, mRNA for E2(14k) and E3alpha (but not E1) were elevated 2-fold in muscles from diabetic rats, although no significant increase in E2(14k) and E3alpha content could be detected by immunoblot or activity assays. The simplest interpretation of these results is that small increases in both E2(14k) and E3alpha in muscles of insulin-deficient animals together accelerate Ub conjugation and protein degradation by the N-end rule pathway, the same pathway activated in cancer cachexia, sepsis, and hyperthyroidism.

  15. The N-end rule pathway catalyzes a major fraction of the protein degradation in skeletal muscle

    NASA Technical Reports Server (NTRS)

    Solomon, V.; Lecker, S. H.; Goldberg, A. L.

    1998-01-01

    In skeletal muscle, overall protein degradation involves the ubiquitin-proteasome system. One property of a protein that leads to rapid ubiquitin-dependent degradation is the presence of a basic, acidic, or bulky hydrophobic residue at its N terminus. However, in normal cells, substrates for this N-end rule pathway, which involves ubiquitin carrier protein (E2) E214k and ubiquitin-protein ligase (E3) E3alpha, have remained unclear. Surprisingly, in soluble extracts of rabbit muscle, we found that competitive inhibitors of E3alpha markedly inhibited the 125I-ubiquitin conjugation and ATP-dependent degradation of endogenous proteins. These inhibitors appear to selectively inhibit E3alpha, since they blocked degradation of 125I-lysozyme, a model N-end rule substrate, but did not affect the degradation of proteins whose ubiquitination involved other E3s. The addition of several E2s or E3alpha to the muscle extracts stimulated overall proteolysis and ubiquitination, but only the stimulation by E3alpha or E214k was sensitive to these inhibitors. A similar general inhibition of ubiquitin conjugation to endogenous proteins was observed with a dominant negative inhibitor of E214k. Certain substrates of the N-end rule pathway are degraded after their tRNA-dependent arginylation. We found that adding RNase A to muscle extracts reduced the ATP-dependent proteolysis of endogenous proteins, and supplying tRNA partially restored this process. Finally, although in muscle extracts the N-end rule pathway catalyzes most ubiquitin conjugation, it makes only a minor contribution to overall protein ubiquitination in HeLa cell extracts.

  16. Acylated triterpene saponins from the roots of Securidaca longepedunculata.

    PubMed

    Mitaine-Offer, Anne-Claire; Pénez, Nicolas; Miyamoto, Tomofumi; Delaude, Clément; Mirjolet, Jean-François; Duchamp, Olivier; Lacaille-Dubois, Marie-Aleth

    2010-01-01

    Four triterpene saponins, 3-O-beta-D-glucopyranosylpresenegenin 28-O-beta-D-apiofuranosyl-(1-->3)-beta-d-xylopyranosyl-(1-->4)-[beta-D-apiofuranosyl-(1-->3)]-alpha-L-rhamnopyranosyl-(1-->2)-{4-O-[(E)-3,4,5-trimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosylpresenegenin 28-O-beta-D-apiofuranosyl-(1-->3)-beta-D-xylopyranosyl-(1-->4)-[beta-D-apiofuranosyl-(1-->3)]-alpha-L-rhamnopyranosyl-(1-->2)-[(6-O-acetyl)-beta-D-glucopyranosyl-(1-->3)]-{4-O-[(E)-3,4,5-trimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, 3-O-beta-D-glucopyranosylpresenegenin 28-O-beta-D-apiofuranosyl-(1-->3)-beta-D-xylopyranosyl-(1-->4)-[beta-D-apiofuranosyl-(1-->3)]-alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-galactopyranosyl-(1-->3)]-{4-O-[(E)-3,4,5-trimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, and 3-O-beta-D-glucopyranosylpresenegenin 28-O-beta-D-apiofuranosyl-(1-->3)-[alpha-L-arabinopyranosyl-(1-->4)]-beta-D-xylopyranosyl-(1-->4)-[beta-D-apiofuranosyl-(1-->3)]-alpha-L-rhamnopyranosyl-(1-->2)-{4-O-[(E)-3,4,5-trimethoxycinnamoyl]}-beta-D-fucopyranosyl ester, were isolated from the roots of Securidaca longepedunculata, together with three known compounds. Their structures were established mainly by 2D NMR techniques and mass spectrometry. 2009 Elsevier Ltd. All rights reserved.

  17. Human hepatoblastoma cells (HepG2) and rat hepatoma cells are defective in important enzyme activities in the oxidation of the C27 steroid side chain in bile acid formation.

    PubMed

    Farrants, A K; Nilsson, A; Pedersen, J I

    1993-12-01

    We have examined the ability of HepG2 human hepatoblastoma cells and 7800 C1 Morris rat hepatoma cells to convert 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) and 3 alpha, 7 alpha-dihydroxy-5 beta-cholestanoic acid (DHCA) to cholic acid and chenodeoxycholic acid, respectively. Cell extracts from both these cell lines could neither form cholic acid from THCA nor from the activated form, THCA-CoA. This suggests that both cell lines are defective in two enzyme activities involved in the pathway, the microsomal THCA-CoA ligase and the peroxisomal THCA-CoA oxidase. Furthermore, we show that the subsequent enzymes are active in the conversion to bile acids, because the product of the THCA-CoA oxidase, 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholest-24-enoyl-coenzyme A (delta 24-THCA-CoA) or delta 24-THCA in the presence of THCA-CoA ligase, are converted to cholic acid by both cell lines. HepG2 cells were able to slowly form chenodeoxycholic acid and cholic acid from 5 beta-cholestane-3 alpha, 7 alpha-diol and 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol, respectively, in 24- and 96-h incubations. The rate of cholic acid formation was lower than the rate for chenodeoxycholic acid and there was a clear accumulation of THCA. 7800 C1 Morris cells had no ability to form cholic acid or chenodeoxycholic acid after 96 h incubation. We conclude that these two cell lines have defects in two enzyme activities involved in the peroxisomal oxidation in bile acid formation, the microsomal THCA-CoA ligase and the peroxisomal THCA-CoA oxidase.

  18. Quaternary organization of the goodpasture autoantigen, the alpha 3(IV) collagen chain. Sequestration of two cryptic autoepitopes by intrapromoter interactions with the alpha4 and alpha5 NC1 domains.

    PubMed

    Borza, Dorin-Bogdan; Bondar, Olga; Todd, Parvin; Sundaramoorthy, Munirathinam; Sado, Yoshikazu; Ninomiya, Yoshifumi; Hudson, Billy G

    2002-10-18

    Goodpasture's (GP) disease is caused by autoantibodies that target the alpha3(IV) collagen chain in the glomerular basement membrane (GBM). Goodpasture autoantibodies bind two conformational epitopes (E(A) and E(B)) located within the non-collagenous (NC1) domain of this chain, which are sequestered within the NC1 hexamer of the type IV collagen network containing the alpha3(IV), alpha4(IV), and alpha5(IV) chains. In this study, the quaternary organization of these chains and the molecular basis for the sequestration of the epitopes were investigated. This was accomplished by physicochemical and immunochemical characterization of the NC1 hexamers using chain-specific antibodies. The hexamers were found to have a molecular composition of (alpha3)(2)(alpha4)(2)(alpha5)(2) and to contain cross-linked alpha3-alpha5 heterodimers and alpha4-alpha4 homodimers. Together with association studies of individual NC1 domains, these findings indicate that the alpha3, alpha4, and alpha5 chains occur together in the same triple-helical protomer. In the GBM, this protomer dimerizes through NC1-NC1 domain interactions such that the alpha3, alpha4, and alpha5 chains of one protomer connect with the alpha5, alpha4, and alpha3 chains of the opposite protomer, respectively. The immunodominant Goodpasture autoepitope, located within the E(A) region, is sequestered within the alpha3alpha4alpha5 protomer near the triple-helical junction, at the interface between the alpha3NC1 and alpha5NC1 domains, whereas the E(B) epitope is sequestered at the interface between the alpha3NC1 and alpha4NC1 domains. The results also reveal the network distribution of the six chains of collagen IV in the renal glomerulus and provide a molecular explanation for the absence of the alpha3, alpha4, alpha5, and alpha6 chains in Alport syndrome.

  19. Enzymic alpha-galactosylation of a cyclic glucotetrasaccharide derived from alternan.

    PubMed

    Biely, P; Puchart, V; Cote, G L

    2001-06-04

    Alternanase catalyzes the hydrolysis of alternan, an alpha-(1-->3)-alpha-(1-->6)-D-glucan produced by Leuconostoc mesenteroides, resulting in the formation of a cyclic tetramer cyclo -->3)-alpha-D-Glcp-(1-->6)-alpha-D-Glcp-(1-->(2) (cGlc(4)). Two alpha-galactosidases, one from coffee bean and the other produced by a fungus, currently described as Thermomyces lanuginosus, were found to catalyze an efficient 6-O-alpha-D-galactopyranosylation of cGlc(4). The attachment of a nonreducing alpha-D-galactopyranosyl residue to the cGlc(4) molecule opens new possibilities for future applications of the cyclic tetramer, since the D-galactopyranosyl residue can be easily modified by D-galactose oxidase to introduce a reactive aldehyde group. The results also extend our knowledge about the synthetic potential of T. lanuginosus alpha-galactosidase.

  20. Novel epimerization of steroidal allylic alcohols.

    PubMed

    Mori, M; Ikegami, S; Tamaoki, B

    1979-04-01

    Epimerization of 4-pregnene-3 beta,20 alpha-diol into 4-pregnene-3 alpha,20 alpha-diol was achieved under the mild condition of an acidic medium at room temperature. This reaction was favorable for synthesis of 4-pregnene-3 alpha,20 alpha-diol in better yield, after chemical reduction of 20 alpha-hydroxy-4-pregnen-3-one with metal hydrides, which resulted in predominant production of 4-pregnene-3 beta,20 alpha-diol. The by-product which was formed more by raising the temperature was identified as 3,5-pregnadien-20 alpha-ol. This method was also applicable for epimerization of other delta 4-3 beta-hydroxysteroids.

  1. [Chemical constituents of Rhododendron seniavinii].

    PubMed

    Wang, Qing-Qing; Zhang, Ying; Ye, Wen-Cai; Zhou, Guang-Xiong

    2013-02-01

    To study the chemical constituents of Rhododendron seniavinii. Compounds were isolated from the aqueous extract of the leaves of R. seniavinii by using Sephadex LH-20, ODS open column chromatography and other means. Their structures were elucidated according to spectral data and physiochemical properties. Thirteen compounds were isolated from R. seniavinii and identified as 5-methoxydehydroconiferyl alcohol (1), dehydroconiferyl alcohol (2), (-)-syringaresinol (3), (-)-lyoniresinol (4), (+)-lyoniresinol 3alpha-O-beta-D-glucopyranoside (5), (-)-lyoniresinol 3alpha-O-beta-D-glucopyranoside (6), 3,4,5-trimethoxyphenyl-1-O-beta-D-glucopyranoside (7), nikoenoside (8), 3,5,7-trihydroxychromone-3-0-alpha-L-rhamnopyranoside (9), 3,4,5-trimethoxyphenol (10), scopoletin (11), scopolin (12) and quercitrin (13). Compounds 1-12 were obtained from this plant for the first time.

  2. Chemical composition of the stem bark and leaves of Ficus pandurata Hance.

    PubMed

    Ramadan, M A; Ahmad, A S; Nafady, A M; Mansour, A I

    2009-01-01

    A new compound, 3-O-alpha-L-arabinopyranosyl-4-hydroxybenzoic acid (13), in addition to 16 newly reported compounds: alpha-amyrin acetate (1), beta-amyrone (2), 3beta-acetoxy-20-taraxasten-22-one (3), alpha-amyrin (4), ceryl alcohol (5), stigmasterol (6), beta-sitosterol (7), 2alpha,3alpha-dihydroxy-lup-20(29)-en-28-oate (8), ursolic acid (9), beta-sitosterol-3-O-glucosoide (10), protocatechuic acid (11), betulinic acid (12), quercetin (14), quercetin-3-O-beta-D-glucoside (15), kampferol-3-O-beta-neohesperidoside (16) and rutin (17) were isolated from the stem bark and leaves of Ficus pandurata (Hance) cultivated in Egypt. Identification of these compounds has been established by physical, chemical and spectral data (UV, IR, MS, (1)H- and (13)C-NMR), as well as comparison with authentic samples.

  3. Antiviral sulfated steroids from the ophiuroid Ophioplocus januarii.

    PubMed

    Roccatagliata, A J; Maier, M S; Seldes, A M; Pujol, C A; Damonte, E B

    1996-09-01

    One new and three known sulfated steroidal polyols have been isolated from the ophiuroid Ophioplocus januarii, collected at San Antonio Oeste, Río Negro, Argentina. The four compounds possess 4 alpha,11 beta-dihydroxy-3 alpha,21-disulfoxy substituents and the A/B cis ring junction but differ in the side chain. The new compound has been characterized as (22E)-5 beta-24-norcholest-22-ene-3 alpha,4 alpha,11 beta,21-tetrol 3,21-disulfate (4). The structures of the four compounds were determined from spectral data and comparison with those of related steroidal polyols. The four compounds were tested for their inhibitory effect on the replication of one DNA and three RNA viruses. Compounds 2 and 4 were active against respiratory syncytial and polio viruses, and compound 3 inhibited Junin virus, responsible for Argentine hemorrhagic fever.

  4. Mercapto steroids in protection against mercury and lead poisoning.

    PubMed

    Blickenstaff, R T; Cox, B; Foster, E; Roberts, L; Steinrauf, L K

    1980-05-01

    When thiocholesterol is administered as liposomes, it provides significant protection against methylmercuric chloride in mice when given in three intraperitoneal injections, 0.5 hr before and 2 and 8 hr after the methylmercuric chloride. Thiositosterol, 5 alpha-cholestane-2 beta, 3 alpha-dithiol, and 5 beta-cholane-3 beta, 24-dithiol also are active, but 3 alpha-mercapto-5 alpha-pregnan-20-one, 6 beta-mercapto-5 alpha-cholestane-3 beta, 5 alpha-diol, 3 beta-mercapto-5 beta-cholanic acid, and adamantanethiol are ineffective under these conditions. Adamantanethiol is somewhat effective when administered in soybean oil. Cholestanyl amine was treated with acetylthiosuccinic anhydride to give the half amide; cleavage with hydroxylamine liberated the thiol group. This product is active against both methylmercuric chloride and lead nitrate.

  5. Structures of acetylated oleanane-type triterpene saponins, rarasaponins IV, V, and VI, and anti-hyperlipidemic constituents from the pericarps of Sapindus rarak.

    PubMed

    Asao, Yasunobu; Morikawa, Toshio; Xie, Yuanyuan; Okamoto, Masaki; Hamao, Makoto; Matsuda, Hisashi; Muraoka, Osamu; Yuan, Dan; Yoshikawa, Masayuki

    2009-02-01

    The methanolic extract and its saponin fraction (methanol-eluted fraction) of the pericarps of Sapindus rarak DC. were found to suppress plasma triglyceride elevation in olive oil-treated mice. From the active fraction, three new acylated oleanane-type triterpene saponins, rarasaponins IV (1), V (2), and VI (3), were isolated. The structures of 1-3 were elucidated on the basis of chemical and spectroscopic evidence. The principle saponin constituents, hederagenin 3-O-alpha-L-arabinopyranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (4) and hederagenin 3-O-(3,4-di-O-acetyl-alpha-L-arabinopyranosyl)-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside (5), showed inhibitory effects on plasma triglyceride elevation at a dose of 200 mg/kg, per os.

  6. Inhibition of pyridine-nucleotide-dependent enzymes by pyrazoles. Synthesis and enzymology of a novel A-ring pyrazole steroid.

    PubMed

    Holt, D A; Levy, M A; Brandt, M; Metcalf, B W

    1986-01-01

    A novel A-ring pyrazole steroid, 2,3-bisaza-A-nor-1,5(10)-estradien-17 beta-ol (3), was synthesized as a potential inhibitor of steroidal NAD(P)H-dependent oxidoreductases. Compound 3 proved to be a potent inhibitor of 3(17)beta-hydroxysteroid dehydrogenase (from P. testosteroni) exhibiting a Ki of 90 +/- 20 nM. The activities of 3 alpha,20 beta-hydroxysteroid dehydrogenase (from S. hydrogenans), steroid-5 alpha-reductase (from rat prostate), and 3 alpha-hydroxysteroid dehydrogenase (from rat liver) were unaffected by pyrazole 3. Dead end inhibition studies indicate an ordered binding of cofactor prior to substrate or pyrazole inhibitor.

  7. [Method of analysis for musk, artificially bred musk and original musk by HPLC].

    PubMed

    Wu, Y; Wu, P; Xiao, X; Yu, D Q

    1989-01-01

    The ether extract of musk, artificially bred musk and original musk reacted with 2,4-dinitrophenylhydrazine to form corresponding phenylhydrazones. The products were determined by HPLC. The conditions of quantitative analysis of muscone hydrazone and qualitative analysis of delta 5-androstene-3 beta-hydroxy-17-one hydrazone and 3 alpha-hydroxy-5 alpha-androstan-17-one hydrazone, such as the choice of chromatographic columns and mobile phase were studied. The results of standard curve and recovery experiment are satisfactory. Dual detection and MS were used to identify and analyse the purity of the chromatographic peak of muscone hydrazone. Experiments of addition of standards were used to confirm the peaks of delta 5-androstene-3 beta-hydroxy-17-one hydrazone and 3 alpha-hydroxy-5 alpha-androstan-17-one hydrazone.

  8. Structure of the acidic O-specific polysaccharide from Proteus vulgaris O39 containing 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-non-2-ulosonic acid.

    PubMed

    Kondakova, A N; Perepelov, A V; Bartodziejska, B; Shashkov, A S; Senchenkova, S N; Wykrota, M; Knirel, Y A; Rozalski, A

    2001-07-12

    The O-specific polysaccharide of Proteus vulgaris O39 was found to contain a new acidic component of Proteus lipopolysaccharides, 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-non-2-ulosonic acid (di-N-acetylpseudaminic acid, Pse5Ac7Ac). The following structure of the polysaccharide was determined by NMR spectroscopy, including 2D 1H,(1)H COSY, TOCSY, ROESY, and 1H,(13)C HMQC experiments, along with selective cleavage of the polysaccharide by solvolysis with anhydrous trifluoromethanesulfonic (triflic) acid: -->8)-beta-Psep5Ac7Ac-(2-->3)-alpha-L-FucpNAc-(1-->3)-alpha-D-GlcpNAc-(1--> The structure established is unique among the O-specific polysaccharides, which is in accordance with classification of the strain studied into a separate Proteus serogroup.

  9. Primordial lithium - New reaction rates, new abundances, new constraints

    NASA Technical Reports Server (NTRS)

    Kawano, Lawrence; Schramm, David; Steigman, Gary

    1988-01-01

    Newly measured nuclear reaction rates for H-3(alpha, gamma)Li-7 (higher than previous values) and Li-7(p, alpha)He-4 (lower than previous values) are shown to increase the Li-7 yield from big band nucleosynthesis for lower baryon-to-photon ratio (less than about 4 x 10 to the 10th). Recent revisions in the He-3(alpha, gamma)Be-7 and the D(p, gamma)He-3 rates enhance the high (greater than 4 x 10 to the 10th) Li-7(Be) production. New, independent determinations of Li abundances in extreme population II stars are in excellent agreement with the work of Spites and give continued confidence in the use of Li-7 in big bang baryon density determinations.

  10. Essential oil from fruit of Peucedanum tauricum Bieb.

    PubMed

    Bartnik, Magdalena; Głowniak, Kazimierz; Mardarowicz, Marek

    2002-01-01

    Essential oil from fruit of Peucedanum tauricum Bieb, was qualitatively and quantitatively investigated. The content of oil determined by distillation with water and m-xylene was 2.2% of dry mass. Gas chromatography (GC) with MS detection and flame ionisation detection (FID) showed that the oil contains 28 compounds (above 99% of sesquiterpenes), of which 9 were identified as beta-elemene (0.6%), beta-caryophyllene (0.3%), alpha-guaiene (0.2%), alpha-humulene (0.8%), gamma-gurjunene (5.6%), beta-selinene (2.3%), alpha-selinene (2.2%), gamma-cadinene (0.3%). Predominating sesquiterpenoids (RI 1529--35.9%, RI 1526--27.2%, RI 1537--7.1%) were not identified, and their mass spectra were similar to mass spectra of selinene derivatives.

  11. Three minor novel triterpenoids from the leaves of Diospyros kaki.

    PubMed

    Chen, Guang; Wang, Zong-Quan; Jia, Ji-Ming

    2009-05-01

    Two novel 18, 19-secoursane triterpenoids, kakisaponin B (1) and kakisaponin C (2), an ursane type 28-nortriterpene, kakidiol (3) and one known triterpenoid rosamultin (4), were isolated from the leaves of Diospyros kaki. The structures of compounds 1 and 2 were determined as 28-O-beta-D-glucopyranosyl-3alpha,19,24-trihydoxy-18,19-secours-11,13(18)-dien-28-oic acid (1) and 28-O-beta-D-glucopyranosyl-2alpha,3alpha,19-trihydoxy-18,19-secours-11,13(18)-dien-28-oic acid (2) by chemical methods and spectra experiments. Kakidiol (3) was characterized as a C(29)-triterpene with an aromatic E-ring in structure. This is the first report of 18,19-secoursane triterpenoids and 28-nortriterpene from family Ebenaceae.

  12. Microbial transformations of testosterone to 5alpha-dihydrotestosterone by two species of Penicillium: P. chrysogenum and P. crustosum.

    PubMed

    Cabeza, M S; Gutiérrez, E B; García, G A; Avalos, A H; Hernández, M A

    1999-06-01

    Two species of Penicillium--P. chrysogenum and P. crustosum--were cultured in presence of [3H]testosterone as a substrate. Both species were shown to reduce the 4,5-double bond in testosterone to give dihydrotestosterone (DHT). The steroids produced were 5alpha-dihydrotestosterone, DHT, 3alpha-hydroxy-5beta-androstan-17-one, 3alpha-hydroy-5alpha-androstan-17-one, 4-androstene-3,17-dione, and 5alpha-androstane-3,17-dione. These products implicate the presence of the 5alpha-reductase, with maximal activity at pH 6 and 8, in both species of Penicillium. The presence of DHT in the growth medium and not in the mycelium suggests that DHT is excreted into the medium.

  13. 3D-graphite structure

    SciTech Connect

    Belenkov, E. A. Ali-Pasha, V. A.

    2011-01-15

    The structure of clusters of some new carbon 3D-graphite phases have been calculated using the molecular-mechanics methods. It is established that 3D-graphite polytypes {alpha}{sub 1,1}, {alpha}{sub 1,3}, {alpha}{sub 1,5}, {alpha}{sub 2,1}, {alpha}{sub 2,3}, {alpha}{sub 3,1}, {beta}{sub 1,2}, {beta}{sub 1,4}, {beta}{sub 1,6}, {beta}{sub 2,1}, and {beta}{sub 3,2} consist of sp{sup 2}-hybridized atoms, have hexagonal unit cells, and differ in regards to the structure of layers and order of their alternation. A possible way to experimentally synthesize new carbon phases is proposed: the polymerization and carbonization of hydrocarbon molecules.

  14. Appearance of cluster states in {sup 13}C

    SciTech Connect

    Yoshida, T.; Itagaki, N.; Otsuka, T.

    2009-03-15

    We study the structure of low-lying states of {sup 13}C with a microscopic cluster model. In addition to the 3{alpha}-n model space, the breaking effect of one of the {alpha} clusters due to the spin-orbit interaction is also taken into account. The isoscalar E0 transition probabilities from the ground 1/2{sup -} state to the excited ones are discussed associated with the cluster structure of these states.

  15. Comparison of osteoblast-like cell responses to calcium silicate and tricalcium phosphate ceramics in vitro.

    PubMed

    Ni, Siyu; Chang, Jiang; Chou, Lee; Zhai, Wanyin

    2007-01-01

    Calcium silicate ceramics have been proposed as new bone repair biomaterials, since they have proved to be bioactive, degradable, and biocompatible. Beta-tricalcium phosphate ceramic is a well-known degradable material for bone repair. This study compared the effects of CaSiO3 (alpha-, and beta-CaSiO3) and beta-Ca3(PO4)2 (beta-TCP) ceramics on the early stages of rat osteoblast-like cell attachment, proliferation, and differentiation. Osteoblast-like cells were cultured directly on CaSiO3 (alpha-, and beta-CaSiO3) and beta-TCP ceramics. Attachment of a greater number of cells was observed on CaSiO3 (alpha-, and beta-CaSiO3) ceramics compared with beta-TCP ceramics after incubation for 6 h. SEM observations showed an intimate contact between cells and the substrates, significant cells adhesion, and that the cells spread and grew on the surfaces of all the materials. In addition, the proliferation rate and alkaline phosphatase (ALP) activity of the cells on the CaSiO3 (alpha-, and beta-CaSiO3) ceramics were improved when compared with the beta-TCP ceramics. In the presence of CaSiO3, elevated levels of calcium and silicon in the culture medium were observed throughout the 7-day culture period. In conclusion, the results of the present study revealed that CaSiO3 ceramics showed greater ability to support cell attachment, proliferation, and differentiation than beta-TCP ceramic. 2006 Wiley Periodicals, Inc.

  16. An analysis of the metabolites of progesterone produced by isolated Sertoli cells at the onset of gametogenesis.

    PubMed

    Wiebe, J P; Tilbe, K S; Buckingham, K D

    1980-05-01

    Sertoli cells isolated from 17 day old rats were maintained in culture and incubated with [14C]-progesterone for 20 h. The cells and media were extracted with ether/chloroform and the extracts chromatographed two-dimensionally on TLC and the radioactive metabolites visualized by autoradiography. Nine of the metabolites (constituting about 88% of total metabolite radioactivity) were identified by relative mobilities of the compounds and their derivatives in TLC and GC systems and by recrystallizations with authentic steroids as the following: 20 alpha-hydroxypregn-4-en-3-one, 3 alpha-hydroxy-5 alpha-pregnan-20-one, 5 alpha-pregnane-3 alpha, 20 alpha-diol, 17 beta-hydroxy-5 alpha-androstan-3-one, 5 alpha-pregnane-3,20-dione, 17-hydroxypregn-4-ene-3,20-dione, testosterone, 5 alpha-androstane-3 alpha, 17 beta-diol and androst-4-ene-3,17-dione. Over 71% of the metabolite radioactivity was due to 20 alpha-hydroxypregn-4-en-3-one, the major metabolite. 5 alpha-reduced pregnanes constituted about 12% and C19 steroids comprised about 2.9% of the radioactivity of the metabolites. Calculation of relative steroidogenic enzyme activities from initial reaction rates suggested the following activities in muunits/mg Sertoli cell protein: 20 alpha-hydroxysteroid oxidoreductase (20 alpha-HSO; 7.71), 5 alpha-reductase (4.77), 3 alpha-HASO (3.57), 17 alpha-hydroxylase (0.93), 17 beta-HSO (0.34) and C17-C20 lyase (0.34). The relatively high rate of steroidogenic enzyme activities in the Sertoli cells of young rats may indicate that Sertoli cells are less dependent on Leydig cell steroidogenesis than has been assumed. Since nearly all the metabolites of progesterone and testosterone are now identified, it is possible to construct a picture of Sertoli cell steroidogenic activity.

  17. Determination of marker constituents from Cissus quadrangularis Linn. and their quantitation by HPTLC and HPLC.

    PubMed

    Mehta, M; Kaur, N; Bhutani, K K

    2001-01-01

    Four marker constituents, namely, onocer-7-ene-3 alpha, 21 beta-diol, delta-amyrin, delta-amyrone and 3,3',4,4'-tetrahydroxybiphenyl of an Ayurvedic crude drug Cissus quadrangularis Linn. are defined for standardisation purposes. 3,3',4,4'-Tetrahydroxybiphenyl has been isolated for the first time from this drug. The contents of the marker constituents were quantitatively determined by HPTLC and HPLC methods in samples collected from five different geographic zones of India.

  18. Quality experimental and calculated powder x-ray diffraction

    SciTech Connect

    Sullenger, D.B.; Cantrell, J.S.; Beiter, T.A.; Tomlin, D.W.

    1996-08-01

    For several years, we have submitted quality powder XRD patterns to the International Centre for Diffraction Data for inclusion as reference standards in their Powder Diffraction File. The procedure followed is described; examples used are {beta}-UH{sub 3}, {alpha}- BaT{sub 2}, alpha-lithium disilicate ({alpha}-Li{sub 2}Si{sub 2}O{sub 5}), and 2,2`,4,4`,6,6`hexanitroazobenzene-III (HNAB-III).

  19. The inner ear of dogs with X-linked nephritis provides clues to the pathogenesis of hearing loss in X-linked Alport syndrome.

    PubMed

    Harvey, S J; Mount, R; Sado, Y; Naito, I; Ninomiya, Y; Harrison, R; Jefferson, B; Jacobs, R; Thorner, P S

    2001-09-01

    Alport syndrome is an inherited disorder of type IV collagen with progressive nephropathy, ocular abnormalities, and high-tone sensorineural deafness. In X-linked Alport syndrome, mutations in the COL4A5 gene encoding the alpha5 chain of type IV collagen lead to loss of the alpha3/alpha4/alpha5 network and increased susceptibility of the glomerular basement membrane to long-term damage. The molecular defects that underlie the otopathology in this disease remain poorly understood. We used a canine model of X-linked Alport syndrome to determine the expression of type IV collagen alpha-chains in the inner ear. By 1 month in normal adult dogs, the alpha3, alpha4, and alpha5 chains were co-expressed in a thin continuous line extending along the basilar membrane and the internal and external sulci, with the strongest expression along the lateral aspect of the spiral ligament in the basal turn of the cochlea. Affected dogs showed complete absence of the alpha3/alpha4/alpha5 network. The lateral aspect of the spiral ligament is populated by tension fibroblasts that express alpha-smooth muscle actin and nonmuscle myosin and are postulated to generate radial tension on the basilar membrane via the extracellular matrix for reception of high frequency sound. We propose that in Alport syndrome, the loss of the alpha3/alpha4/alpha5 network eventually weakens the interaction of these cells with their extracellular matrix, resulting in reduced tension on the basilar membrane and the inability to respond to high frequency sounds.

  20. [Chemical constituents of Myristica fragrans Houttuyn seed and their physiological activities].

    PubMed

    Maeda, Aki; Tanimoto, Shinichi; Abe, Tomo; Kazama, Shunsuke; Tanizawa, Hisayuki; Nomura, Masato

    2008-01-01

    The antioxidative activity of phenylpropanoid compound extracts from nutmeg (Myristica fragragrans Houttuyn) seed was determined. The antioxidant activity was evaluated using the 1,1-diphenyl-2-picrylhydrazyl radical-scavenging method, superoxide disumutase assay, ferric thiocyanate assay, and radical-scavenging effect assay with electron-spin resonance. High antioxidant activity was found in monoterpenoid extracts including terpinene-4-ol (3), alpha-terpineol (4), and 4-allyl-2,6-dimethoxyphenol (12). Compound, 12 expressed particularly high antioxidant activity.

  1. Withdrawal properties of a neuroactive steroid: implications for GABA(A) receptor gene regulation in the brain and anxiety behavior.

    PubMed

    Smith, Sheryl S

    2002-05-01

    Early work in the field established that the 5 alpha-reduced metabolite of progesterone 3 alpha-OH-5 alpha-pregnan-20-one (allopregnanolone or 3 alpha,5 alpha-THP) is a potent positive modulator of the GABA(A) receptor (GABAR), the receptor mediating the effects of the primary inhibitory transmitter in the brain. This steroid acts in a manner similar to sedative drugs, such as the barbiturates, both in terms of potentiating GABA-induced inhibition in vitro and in behavioral assays, by reducing anxiety and seizure susceptibility. Because sedative compounds exhibit withdrawal properties that result in behavioral hyperexcitability, our laboratory has more recently investigated the effect of prolonged application and rapid removal (i.e. 'withdrawal') of this steroid, administered in vivo to female rats. Withdrawal from 3 alpha,5 alpha-THP produces a state of increased anxiety and lowered seizure threshold, similar to withdrawal from other GABA-modulatory drugs such as the benzodiazepines and alcohol. Hormone withdrawal also produced increases in the alpha 4-containing GABAR, an effect correlated with insensitivity of the GABAR to modulation by the benzodiazepine class of tranquilizers, as would normally occur under control conditions. In addition, changes in intrinsic channel properties, including a marked acceleration in the decay rate was also observed as a result of declining levels of 3 alpha,5 alpha-THP. Such a change would result in less inhibitory total current, and the resulting increase in neuronal excitability could then underlie the observed behavioral excitability following hormone withdrawal. These results suggest that actions of this steroid on a traditional transmitter receptor in the brain lead to alterations in GABAR subunit composition that result in changes in the intrinsic channel properties of the receptor and behavioral excitability. These results may have implications for endogenous fluctuations in this hormone which may accompany premenstrual

  2. Individual Differences in Cerebral Cortical Activity During Stress: Understanding and Intervention to Enhance Shooting Performance

    DTIC Science & Technology

    2009-04-30

    successfully raised physiological and 15. SUBJECT TERMS brain, cognitive neuroscience, EEG , neurofeedback , competition, stress, neuroendocrine, shooting...efficacy of the Neurofeedback training to elevate frontal EEG asymmetry (F4 minus F3 alpha power) in an attempt to enhance emotion regulation. The...observed a remarkable increase or synchrony of EEG alpha power (i.e., low-alpha) across the general scalp topography for both groups ( neurofeedback

  3. In vivo and vitro studies on formation of bile acids in patients with Zellweger syndrome. Evidence that peroxisomes are of importance in the normal biosynthesis of both cholic and chenodeoxycholic acid.

    PubMed Central

    Kase, B F; Pedersen, J I; Strandvik, B; Björkhem, I

    1985-01-01

    The last step in bile acid formation involves conversion of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) into cholic acid and 3 alpha,7 alpha-dihydroxy-5 beta-cholestanoic acid (DHCA) into chenodeoxycholic acid. The peroxisomal fraction of rat and human liver has the highest capacity to catalyze these reactions. Infants with Zellweger syndrome lack liver peroxisomes, and accumulate 5 beta-cholestanoic acids in bile and serum. We recently showed that such an infant had reduced capacity to convert a cholic acid precursor, 5 beta-cholestane-3 alpha,7 alpha,12 alpha-triol into cholic acid. 7 alpha-Hydroxy-4-cholesten-3-one is a common precursor for both cholic acid and chenodeoxycholic acid. Intravenous administration of [3H]7 alpha-hydroxy-4-cholesten-3-one to an infant with Zellweger syndrome led to a rapid incorporation of 3H into biliary THCA but only 10% of 3H was incorporated into cholic acid after 48 h. The incorporation of 3H into DHCA was only 25% of that into THCA and the incorporation into chenodeoxycholic acid approximately 50% of that in cholic acid. The conversion of intravenously administered [3H]THCA into cholic acid in another infant with Zellweger syndrome was only 7%. There was a slow conversion of THCA into 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-C29-dicarboxylic acid. The pool size of both cholic- and chenodeoxycholic acid was markedly reduced. Preparations of liver from two patients with Zellweger syndrome had no capacity to catalyze conversion of THCA into cholic acid. There was, however, a small conversion of DHCA into chenodeoxycholic acid and into THCA. It is concluded that liver peroxisomes are important both for the conversion of THCA into cholic acid and DHCA into chenodeoxycholic acid. PMID:4077985

  4. Cadinane sesquiterpenes from Curcuma parviflora.

    PubMed

    Sadhu, Samir Kumar; Tamaki, Mayu; Ohtsuki, Takashi; Toume, Kazufumi; Koyano, Takashi; Kowithayakorn, Thaworn; Ishibashi, Masami

    2009-04-01

    Two new cadinane sesquiterpenes (1 and 2) were isolated from Curcuma parviflora, and their structures were elucidated by spectroscopic data. Compound 1, 4alpha-acetoxycadina-2,9-diene-1,8-dione, possesses two conjugated enone chromophores, while compound 2, 1alpha,3alpha,4beta-trihydroxy-9-cadinen-8-one, has an enone moiety with three hydroxy groups. Isolation of these cadinane monomers may reasonably suggest that parviflorenes are biogenetically classified as cadinane dimers.

  5. A new phenolic fatty acid ester with lipoxygenase inhibitory activity from Jacaranda filicifolia.

    PubMed

    Ali, R M; Houghton, P J

    1999-06-01

    The dichloromethane extract of the stem of jacaranda filicifolia Don. showed inhibitory activity in vitro against soybean 5-lipoxygenase. Systematic fractionation to isolate the compounds responsible resulted in the isolation of three active compounds, 2 alpha, 3 alpha-dihydroxyurs-12-en-28-oic acid, beta-sitosterol, and one of which was new which was characterised as 2-(4-hydroxyphenyl)ethyl 1-dodecyloctadecanoate. This type of compound has not previously been reported to inhibit lipoxygenase.

  6. A new 28-noroleanane triterpenoid from the aerial parts of Lantana camara Linn.

    PubMed

    Begum, Sabira; Zehra, Syeda Qamar; Ayub, Anjum; Siddiqui, Bina Shaheen

    2010-08-01

    A new 28-norolean-12,17-diene triterpene lantigdienone (1) oxidised at C-11 and C-22 has been isolated from the aerial parts of Lantana camara, along with two known compounds, camarinin (2) and camangeloyl acid (3). The structure of compound 1 was elucidated as 3,25-epoxy-3alpha-hydroxy-22beta-[beta,betadimethylacryloyloxy]-11-oxo-28-norolean-12,17-diene, with the help of spectral studies.

  7. Natural products from Scorzonera aristata (Asteraceae).

    PubMed

    Jehle, Manuela; Bano, Johanna; Ellmerer, Ernst P; Zidorn, Christian

    2010-05-01

    The aerial parts of Scorzonera aristata Ramond ex DC., collected in the South Tyrolean Dolomites, yielded the flavonoids quercetin 3-O-glucoside, rutin, and isoorientin, and the caffeic acid derivatives chlorogenic acid, 4,5-dicaffeoyl quinic acid, and 3,5-dicaffeoyl quinic acid. Sub-aerial parts contained caffeic acid methyl ester, 3,5-dicaffeoyl quinic acid, and the triterpenes 3alpha-hydroxyolean-5-ene, lupeol, and magnificol. Chemosystematic implications of the isolated compounds are discussed briefly.

  8. Exploiting the CNC side chain in heterocyclic rearrangements: synthesis of 4(5)-acylamino-imidazoles.

    PubMed

    Piccionello, Antonio Palumbo; Buscemi, Silvestre; Vivona, Nicolò; Pace, Andrea

    2010-08-06

    A new variation on the Boulton-Katritzky reaction is reported, namely, involving use of a CNC side chain. A novel Montmorillonite-K10 catalyzed nonreductive transamination of a 3-benzoyl-1,2,4-oxadiazole afforded a 3-(alpha-aminobenzyl)-1,2,4-oxadiazole, which was condensed with benzaldehydes to afford the corresponding imines. In the presence of strong base, these imines underwent Boulton-Katritzky-type rearrangement to afford novel 4(5)-acylaminoimidazoles.

  9. Brain testosterone metabolism in thyroidectomized and thyroxine-treated chickens.

    PubMed

    Klandorf, H; Lucini, V; Harvey, S

    1984-10-01

    The metabolism of testosterone to reduced derivatives was studied in the pituitary gland, the hypothalamus, and the hyperstriatum dorsale of thyroidectomized, sham-operated, and thyroxine (T4)-injected immature cockerels. The levels of plasma thyroid hormones were markedly reduced (P less than 0.001) in thyroidectomized cockerels whereas thyroidectomized or sham-operated birds injected daily with 100 micrograms/kg thyroxine had significantly elevated (P less than 0.001) levels in comparison with sham-operated control birds. Each tissue was found to produce significant amounts of 5 beta-androstane-17 beta-ol-3-one (5 beta-DHT), 5 beta-androstane-3 alpha, 17 beta-diol (5 beta-3 alpha-diol), and androstenedione. Irrespective of thyroid state 5 beta-DHT and 5 beta-3 alpha-diol were produced to the greatest extent by the hyperstriatum dorsale whereas androstenedione was maximally produced in the pituitary gland. In comparison with the hyperstriatum dorsale and the hypothalamus only small quantities of 5 beta-DHT were produced in the pituitary gland. In the hyperstriatum dorsale of thyroidectomized birds both 5 beta-DHT (P less than 0.05) and 5 beta-3 alpha-diol (P less than 0.1) were formed to a greater extent than in sham-operated birds. This effect was reversed by administration of T4 to the operated birds which reduced the levels to those measured in the sham-operated controls. Similarly, injection of T4 into sham-operated birds decreased (P less than 0.05) the production of 5 beta-DHT in the hypothalamus while in the pituitary gland injection of T4 into thyroidectomized birds reduced the production of androstenedione (P less than 0.05). It was concluded that in the cockerel thyroid hormone is likely to play a role in the metabolism of testosterone. The physiological significance of 5 beta-reductase activity in the neuroendocrine tissues is discussed.

  10. A new secotrinervitane diterpene isolated from soldiers of the Madagascan termite species, Nasutitermes canaliculatus.

    PubMed

    Rabemanantsoa, A; Ranarivelo, Y; Andriantsiferana, M; Tillequin, F; Silverton, J V; Garraffo, H M; Spande, T F; Yeh, H J; Daly, J W

    1996-09-01

    Reported herein is the X-ray crystallographic structure of a novel 10-oxygenated secotrinervitane diterpene, 3 alpha, 10 alpha-diacetoxy-7,16-secotrinervita-7,11,15(17)-triene (4), from soldiers of the endemic Madagascan termite Nasutitermes canaliculatus, which was compared with an energy-minimized structure obtained by computer molecular modeling. We also report 1H- and 13C-NMR and MS data for this new diterpene.

  11. Phytochemical and molluscicidal investigations of Fagonia arabica.

    PubMed

    El-Wakil, Eman A

    2007-01-01

    The aqueous methanolic extract of the aerial parts of Fagonia arabica L. (family Zygophyllaceae) was successively fractionated using certain organic solvents. From the ethyl acetate fraction, two flavonoid glycosides were isolated and identified as kaempferol-7-O-rhamnoside and acacetin-7-O-rhamnoside. Four triterpenoidal glycosides were isolated from the butanolic layer. Their structures were elucidated on the basis of the spectral and chemical data as 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranoside oleanolic acid (1), 3-O-alpha-L-arabinopyranosyl quinovic acid 28-O-beta-D-glucopyranoside (2), 3-O-[beta-D-glucopyranosyl-(1-->2)]-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinosyl oleanolic acid (3) and 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabino-pyranosyl quinovic acid 28-O-beta-D-glucopyranoside (4). The two monodesmosidic saponins 1 and 3 were found to possess strong molluscicidal activity against Biomphalaria alexandrina snails, the intermediate host of Schistosoma mansoni in Egypt (LC90 = 13.33 and 16.44 microM), whereas the other two bidesmosidic saponins 2 and 4 as well as the two flavonoid glycosides were inactive up to 50 microM.

  12. Complete structure of the tyrosine-linked saccharide moiety from the surface layer glycoprotein of Clostridium thermohydrosulfuricum S102-70.

    PubMed Central

    Christian, R; Schulz, G; Schuster-Kolbe, J; Allmaier, G; Schmid, E R; Sleytr, U B; Messner, P

    1993-01-01

    In this study, we have extended and completed a previous investigation (P. Messner, R. Christian, J. Kolbe, G. Schulz, and U. B. Sleytr, J. Bacteriol. 174:2236-2240, 1992) in which we demonstrated for the first time in prokaryotic organisms the presence of a novel O-glycosidic linkage via tyrosine. The surface layer glycoprotein of the eubacterium Clostridium thermohydrosulfuricum S102-70 is arranged in a hexagonal lattice, with center-to-center spacings of approximately 16.3 nm. Molecular weight determination by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of both glycosylated and chemically deglycosylated surface layer glycoprotein showed values for the monomeric subunits of 94,000 and 87,500, respectively. Glycopeptide fractions obtained after exhaustive pronase digestion of purified, intact glycoprotein were isolated by reversed-phase liquid chromatography. One- and two-dimensional nuclear magnetic resonance studies, together with chemical analyses and plasma desorption time-of-flight mass spectrometry, were used to elucidate the structure of the hexasaccharide moiety linked by the novel O-glycosidic linkage to tyrosine. The combined evidence suggests the following structure: beta-D-Galf-(1-->3)-alpha-D-Galp- (1-->2)-alpha-L-Rhap-(1-->3)-alpha-D-Manp-(1--3)-alpha-L- Rhap-(1-->3)-beta- D-Glcp-(1-->4)-L-Tyr. Images PMID:8444787

  13. Gonadotropin stimulation of steroid synthesis and metabolism in the Rana pipiens ovarian follicle: sequential changes in endogenous steroids during ovulation, fertilization and cleavage stages.

    PubMed

    Morrill, Gene A; Schatz, Frederick; Kostellow, Adele; Bloch, Eric

    2006-05-01

    Steroid synthesis and metabolism have been followed in Rana pipiens ovarian follicles, denuded oocytes and eggs during ovulation, fertilization and cleavage stages (blastula formation). Under physiological conditions, gonadotropin stimulation of the fully grown follicle leads to progesterone synthesis from [(3)H]acetate as well as formation of much smaller amounts of 17alpha-hydroxyprogesterone, androstenedione, pregnanedione and pregnanediol. Progesterone levels increase during completion of the first meiotic division, but by ovulation progesterone disappears from the egg. Plasma membrane-bound progesterone is taken up into the oocyte cortical granules and is largely metabolized to 5alpha-pregnane-3alphaol,20-one and 5beta-pregnane-3alpha,17alpha,20beta-triol coincident with internalization of 60% of the oocyte surface (and >90% of bound progesterone) by the end of the hormone-dependent period. The principal steroid in the ovulated egg is 5beta-pregnane-3alpha,17alpha,20beta-triol. There is a rapid efflux of 5beta-pregnane-3alpha,17alpha,20beta-triol into the medium immediately following fertilization and residual steroid levels remain low in the developing blastula. Dissociated blastulae cells prepared from stage 9 1/2 embryos concentrate both pregnenolone and progesterone from the medium with minimal metabolism. The results indicate that the ovarian follicle has the ability to synthesize and metabolize progesterone but that this ability disappears in the ovulated egg. The progesterone metabolites formed during meiosis are largely released at fertilization.

  14. Similarity of "core" structures in two different glycans of tyrosine-linked eubacterial S-layer glycoproteins.

    PubMed Central

    Messner, P; Christian, R; Neuninger, C; Schulz, G

    1995-01-01

    Previously, the repeating-unit structure of the S-layer glycoprotein from the eubacterium Bacillus alvei CCM 2051 has been determined to be [-->3)-beta-D-Galp-(1-->4)-[alpha-D-Glcp-(1-->6)-]-beta-D-ManpNAc- (1-->]n (E. Altman, J.-R. Brisson, P. Messner, and U. B. Sleytr, Biochem. Cell Biol. 69:72-78, 1991). Nuclear magnetic resonance spectroscopic reexamination of this glycan reveals that the O-antigen-like domain of the polysaccharide is [see text] connected with the S-layer polypeptide through the "core" structure -->3)-alpha-L-Rhap-(1-->3)-alpha-L-Rhap-(1-->3)-alpha-L-R hap-(1-->3)-beta-D-Galp-(1-->O)-Tyr. Except for the substitution in position 4 of the nonreducing rhamnose with the modified glyceric acid phosphate residue GroA-2-->OPO2-->4-beta-D-ManpNAc-(1-->, this core is identical to the core of the tyrosine-linked glycan from the S-layer glycoprotein of Thermoanaerobacter thermohydrosulfuricus L111-69 (K. Bock, J. Schuster-Kolbe, E. Altman, G. Allmaier, B. Stahl, R. Christian, U. B. Sleytr, and P. Messner, J. Biol. Chem. 269:7137-7144, 1994). PMID:7721708

  15. Purification and properties of rabconnectin-3.

    PubMed

    Sakisaka, Toshiaki; Takai, Yoshimi

    2005-01-01

    Rab3A, a member of the Rab3 small GTP-binding protein (G protein) family, regulates Ca(2+)-dependent exocytosis of neurotransmitter. The cyclical activation and inactivation of Rab3A are essential for the Rab3A action in exocytosis. GDP-Rab3A is activated to GTP-Rab3A by Rab3 GDP/GTP exchange protein (Rab3 GEP), and GTP-Rab3A is inactivated to GDP-Rab3A by Rab3 GTPase-activating protein (Rab3 GAP). We) have found a novel protein, named rabconnectin-3, that is coimmunoprecipitated with Rab3 GEP or GAP from the crude synaptic vesicle fraction of rat brain. Rabconnectin-3 constitutes a subunit structure consisting of alpha and beta subunits and localizes at synaptic vesicles. Overexpression of the C-terminal fragment of rabconnectin-3alpha inhibits Ca(2+)-dependent exocytosis from PC12 cells. We describe the purification method for native rabconnectin-3alpha and -3beta from rat brain and the functional properties of rabconnectin-3alpha in Ca(2+)-dependent exocytosis by use of human growth hormone coexpression assay system of PC12 cells.

  16. Human C3 and C5: subunit structure and modifications by trypsin and C42-C423.

    PubMed

    Nilsson, U R; Mandle, R J; McConnell-Mapes, J A

    1975-02-01

    The subunit composition of human C3 and C5 was analyzed. Acrylamide gel electrophoresis of the fully reduced and dissociated proteins disclosed a similar structure, consisting of one alpha and beta subunit, linked together by one or more disulfide bonds. The approximate molecular weights for the alpha and beta subunits of C3 as well as C5 were 140,000 and 80,000 respectively. C42 caused cleavage solely of C3alpha, whereas trypsin affected both C3alpha as well as C3beta. A characteristic subunit modification by both enzmes indicated that C3alpha constitutes the source of C3a. C423 as well as trypsin exclusively affect C5alpha. C5a therefore appears to originate from the C5alpha subunit. The mode of primary cleavage by C423 and trypsin differs, giving rise to different forms of C5b. The questions is raised if multiple forms of C5a also exist. It appeared from our studies that certain forms of C5b may retain portions of the alpha subunit, which could potentially release some biologically active split products following secondary cleavage by the appropriate enzyme.

  17. Novel lanostane and rearranged lanostane-type triterpenoids from Abies sachalinensis - II -.

    PubMed

    Gao, Hui Yuan; Wu, Li Jun; Nakane, Takahisa; Shirota, Osamu; Kuroyanagi, Masonori

    2008-04-01

    In the previous work we reported five A-seco-rearranged lanostane triterpenoids as antibacterial constituents from the ethyl acetate soluble fraction of Abies sachalinensei leaves. In further study on the isolation of constituents from the ethyl acetate soluble fraction, two new rearranged lanostane and lanostane-type triterpenoids (3, 4) and three reported compound (1, 2, 5) were isolated. The structures of new compound 3 and 4 were determined to be 3,4-seco-4(28),6,8(14),24-mariesatetraen-26,23-olide-23-hydroxy-3-oic acid and 3,4-seco-4(28),7,24-lanostatrien-26,23-olide-23-hydroxy-3-oic acid, respectively, by spectral studies on HR-MS, (1)H-NMR, (13)C-NMR, and 2D-NMR spectra. Compound 1 was identified with pindrolactone and its structure was revised as 7,14,22Z,24-mariesatetraen-26,23-olide-3alpha-ol. Structures of 2 and 5 were determined as 7,14,24-mariesatrien-26,23-olide-3alpha,23-diol and 3alpha-hydroxy-7,14,24E-mariesatrien-23-oxo-26-oic acid. Of these compounds, 2, 3 and 4 were obtained as lactol tautomer mixtures at gamma-lactone structures of side chains.

  18. Analgesic strategies aimed at stimulating the endogenous production of allopregnanolone

    PubMed Central

    Poisbeau, Pierrick; Keller, Anne Florence; Aouad, Maya; Kamoun, Nisrine; Groyer, Ghislaine; Schumacher, Michael

    2014-01-01

    A growing number of studies indicate that 3-alpha reduced neurosteroids are remarkable analgesics in various pain states. This is the case for allopregnanolone (AP), one of the most potent endogenous positive allosteric modulators of GABAA receptor function. From the pioneering work of Hans Selye, who described the sedative properties of steroids, synthetic compounds resembling the progesterone metabolite AP have been developed. If some of them have been used as anesthetics, it seems difficult to propose them as a therapeutic option for pain since they display several adverse side effects such as sedation, amnesia and functional tolerance. An alternative strategy, chosen by few laboratories around the world, is aimed at stimulating the local production of 3-alpha reduced neurosteroids in order to limit these well-known side effects. This pharmacological approach has the advantage of targeting specific structures, fully equipped with the necessary biosynthetic enzymatic machinery, where neurosteroids already act as endogenous pain modulators. The various pharmacological trials which attempted to treat pain symptoms by stimulating the production of 3-alpha reduced neurosteroids are reviewed here, as well as novel neurotransmitter systems possibly regulating their endogenous production. PMID:24987335

  19. Negative regulation of opioid receptor-G protein-Ca2+ channel pathway by the nootropic nefiracetam.

    PubMed

    Yoshii, Mitsunobu; Furukawa, Taiji; Ogihara, Yoshiyasu; Watabe, Shigeo; Shiotani, Tadashi; Ishikawa, Yasuro; Nishimura, Masao; Nukada, Toshihide

    2004-10-01

    It has recently been reported that nefiracetam, a nootropic agent, is capable of attenuating the development of morphine dependence and tolerance in mice. The mechanism of this antimorphine action is not clear. The present study was designed to address this issue using Xenopus oocytes expressing delta-opioid receptors, G proteins (G(i3alpha) or G(o1alpha)), and N-type (alpha1B) Ca2+ channels. Membrane currents through Ca2+ channels were recorded from the oocytes under voltage-clamp conditions. The Ca2+ channel currents were reduced reversibly by 40-60% in the presence of 1 microM leucine-enkephalin (Leu-Enk). The Leu-Enk-induced current inhibition was recovered promptly by nefiracetam (1 microM), while control currents in the absence of Leu-Enk were not influenced by nefiracetam. A binding assay revealed that 3H-nefiracetam preferentially bound to the membrane fraction of oocytes expressing G(i3alpha). When delta-opioid receptors were coexpressed, the binding was significantly increased. However, an additional expression of alpha1B Ca2+ channels decreased the binding. The results suggest that nefiracetam preferentially binds to G(i3alpha) associated with delta-opioid receptors, thereby inhibiting the association of G proteins with Ca2+ channels. In conclusion, nefiracetam negatively regulates the inhibitory pathway of opioid receptor-G protein-Ca2+ channel.

  20. Metabolism of methyltestosterone in the greyhound.

    PubMed

    Biddle, S T B; O'Donnell, A; Houghton, E; Creaser, C

    2009-03-01

    Gas chromatography/mass spectrometry and selective derivatisation techniques have been used to identify urinary metabolites of methyltestosterone following oral administration to the greyhound. Several metabolites were identified including reduced, mono-, di- and trihydroxylated steroids. The major metabolites observed were 17alpha-methyl-5beta-androstane-3alpha-17beta-diol, 17alpha-methyl-5beta-androstane-3alpha,16alpha,17beta-triol, and a further compound tentatively identified as 17alpha-methyl-5z-androstane-6z,17beta-triol. The most abundant of these was the 17alpha-methyl-5beta-androstane-3alpha,16alpha,17beta-triol. This metabolite was identified by comparison with a reference standard synthesised using a Grignard procedure and characterised using trimethylsilyl (TMS) and acetonide-TMS derivatisation techniques. There did not appear to be any evidence for 16beta-hydroxylation as a phase I metabolic transformation in the greyhound. However, significant quantities of 16alpha-hydroxy metabolites were detected. Selective enzymatic hydrolysis procedures indicated that the major metabolites identified were excreted as glucuronic acid conjugates. Metabolic transformations observed in the greyhound have been compared with those of other mammalian species and are discussed here.

  1. Genetic diversity of Plasmodium vivax in Kolkata, India

    PubMed Central

    Kim, Jung-Ryong; Imwong, Mallika; Nandy, Amitabha; Chotivanich, Kesinee; Nontprasert, Apichart; Tonomsing, Naowarat; Maji, Ardhendu; Addy, Manjulika; Day, Nick PJ; White, Nicholas J; Pukrittayakamee, Sasithon

    2006-01-01

    Background Plasmodium vivax malaria accounts for approximately 60% of malaria cases in Kolkata, India. There has been limited information on the genotypic polymorphism of P. vivax in this malaria endemic area. Three highly polymorphic and single copy genes were selected for a study of genetic diversity in Kolkata strains. Methods Blood from 151 patients with P. vivax infection diagnosed in Kolkata between April 2003 and September 2004 was genotyped at three polymorphic loci: the P. vivax circumsporozoite protein (pvcs), the merozoite surface protein 1 (pvmsp1) and the merozoite surface protein 3-alpha (pvmsp3-alpha). Results Analysis of these three genetic markers revealed that P. vivax populations in Kolkata are highly diverse. A large number of distinguishable alleles were found from three genetic markers: 11 for pvcs, 35 for pvmsp1 and 37 for pvmsp3-alpha. These were, in general, randomly distributed amongst the isolates. Among the 151 isolates, 142 unique genotypes were detected the commonest genotype at a frequency of less than 2% (3/151). The overall rate of mixed genotype infections was 10.6%. Conclusion These results indicate that the P. vivax parasite population is highly diverse in Kolkata, despite the low level of transmission. The genotyping protocols used in this study may be useful for differentiating re-infection from relapse and recrudescence in studies assessing of malarial drug efficacy in vivax malaria. PMID:16907979

  2. Stromal laminin chain distribution in normal, hyperplastic and malignant oral mucosa: relation to myofibroblast occurrence and vessel formation.

    PubMed

    Franz, Marcus; Wolheim, Anke; Richter, Petra; Umbreit, Claudia; Dahse, Regine; Driemel, Oliver; Hyckel, Peter; Virtanen, Ismo; Kosmehl, Hartwig; Berndt, Alexander

    2010-04-01

    The contribution of stromal laminin chain expression to malignant potential, tumour stroma reorganization and vessel formation in oral squamous cell carcinoma (OSCC) is not fully understood. Therefore, the expression of the laminin chains alpha2, alpha3, alpha4, alpha5 and gamma2 in the stromal compartment/vascular structures in OSCC was analysed. Frozen tissue of OSCC (9x G1, 24x G2, 8x G3) and normal (2x)/hyperplastic (11x) oral mucosa was subjected to laminin chain and alpha-smooth muscle actin (ASMA) immunohistochemistry. Results were correlated to tumour grade. The relation of laminin chain positive vessels to total vessel number was assessed by immunofluorescence double labelling with CD31. Stromal laminin alpha2 chain significantly decreases and alpha3, alpha4, alpha5 and gamma2 chains and also ASMA significantly increase with rising grade. The amount of stromal alpha3, alpha4 and gamma2 chains significantly increased with rising ASMA positivity. There is a significant decrease in alpha3 chain positive vessels with neoplastic transformation. Mediated by myofibroblasts, OSCC development is associated with a stromal up-regulation of laminin isoforms possibly contributing to a migration promoting microenvironment. A vascular basement membrane reorganization concerning alpha3 and gamma2 chain laminins during tumour angioneogenesis is suggested.

  3. Steroidogenesis by gonads of a viviparous teleost, the sailfin molly (Poecilia latipinna), in vitro and in vivo.

    PubMed

    Kime, D E; Groves, D J

    1986-07-01

    Gonads of Poecilia latipinna transformed [3H]testosterone into a number of reduced and conjugated metabolites in high yield in vitro. In the male 5 beta-androstane-3 alpha, 17 beta-diol, 5 beta-androstane-3 beta, 17 beta-diol, and their "sulphates" were identified. The only 11-oxygenated androgen detected was a compound tentatively identified as 5 beta-androstane-3 beta, 11 beta, 17 beta-triol. In ovarian incubates androstenedione, 5 beta-androstane-3 alpha, 17 beta-diol and its glucuronide, testosterone glucuronide, and 5 beta-androstane-3 alpha, 17 beta-diol glucuronide were identified. Highest yields of the ovarian glucuronides coincided with the termination of vitellogenesis which may indicate a possible pheromonal role of these conjugates. In vivo plasma levels of estradiol in the female were correlated with vitellogenesis and fell markedly after castration or hypophysectomy. In males the plasma concentrations of testosterone, 11-ketotestosterone and 11 beta-hydroxytestosterone and their conjugates were variable and not apparently correlated with testicular weight, but they were reduced to undetectable levels by castration and hypophysectomy. The results suggest that 5 alpha- and 5 beta-reduced steroids may play a role in the reproductive endocrinology of P. latipinna and that measurements of only the "classical" steroid hormones in this and possibly other species may give only a partial and misleading picture of endocrine changes.

  4. Longitudinal profiling of urinary steroids by gas chromatography/combustion/isotope ratio mass spectrometry: diet change may result in carbon isotopic variations.

    PubMed

    Saudan, Christophe; Kamber, Matthias; Barbati, Giulia; Robinson, Neil; Desmarchelier, Aurélien; Mangin, Patrice; Saugy, Martial

    2006-02-02

    Longitudinal profiling of urinary steroids was investigated by using a gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) method. The carbon isotope ratio of three urinary testosterone (T) metabolites: androsterone, etiocholanolone, 5beta-androstane-3alpha,17beta-diol (5beta-androstanediol) together with 16(5alpha)-androsten-3alpha-ol (androstenol) and 5beta-pregnane-3alpha,20alpha-diol (5beta-pregnanediol) were measured in urine samples collected from three top-level athletes over 2 years. Throughout the study, the subjects were living in Switzerland and were residing every year for a month or two in an African country. (13)C-enrichment larger than 2.5 per thousand was observed for one subject after a 2-month stay in Africa. Our findings reveal that (13)C-enrichment caused by a diet change might be reduced if the stay in Africa was shorter or if the urine sample was not collected within the days after return to Switzerland. The steroids of interest in each sample did not show significant isotopic fractionation that could lead to false positive results in anti-doping testing. In contrast to the results obtained with the carbon isotopic ratio, profiling of urinary testosterone/epitestosterone (T/E) ratios was found to be unaffected by a diet change.

  5. Urinary marker of oral pregnenolone administration.

    PubMed

    Saudan, Christophe; Desmarchelier, Aurélien; Sottas, Pierre-Edouard; Mangin, Patrice; Saugy, Martial

    2005-03-01

    Pregnenolone (PREG) can potentially be abused by athletes to maintain an equilibration of the steroidal environment after sex steroids administrations. Five men volunteers orally ingested 50 mg PREG to determine optimal urinary markers for detection of this steroid. Our findings show that ingestion of PREG has no significant effects on the testosterone/epitestosterone (T/E) and testosterone/luteinizing hormone (T/LH) ratios, whereas variable changes on the carbon isotopic values of three T metabolites: androsterone, etiocholanolone, 5beta-androstane-3alpha,17beta-diol (5beta-androstanediol) together with 16(5alpha)-androsten-3alpha-ol (androstenol) and 5beta-pregnane-3alpha,20alpha-diol (pregnanediol) have been observed. The difference between the carbon isotopic values (delta13C-values) of androstenol and pregnanediol is potentially the most reliable marker of exogenous PREG administration in males. For all subjects, the differences differ by 3.0 per thousand or more over a period of about 10 h and for both of them the detection window for positivity is extended over 40 h.

  6. {sup 12}C formation: A classical quest in new light

    SciTech Connect

    Tengblad, O.; Alcorta, M.; Borge, M. J. G.; Madurga, M.; Perea, A.; Cubero, M.; Fynbo, H. O. U.; Riisager, K.; Kirsebom, O.; Hyldegaard, S.; Jonson, B.; Nyman, G.; Nilsson, T.; Diget, D. G.; Fulton, B.

    2011-10-28

    In this work we have studied the break-up of {sup 12}C following the reactions {sup 10}B({sup 3}He,p{alpha}{alpha}{alpha}) and {sup 11}B({sup 3}He,d{alpha}{alpha}{alpha}). The study was performed at the 5 MV tandem in Madrid. The break-up gives us information on excited states in {sup 12}C from the famous Hoyle state up to an energy of almost 18 MeV. Using a highly segmented experimental set-up the simultaneous detection of the three alpha particles in coincidence with a proton or deuteron respectively made possible a full kinematic reconstruction of the break-up. On the basis of the energies of the 3 {alpha} particles and their angular correlations it has been possible to determine the spin and parity of states for cases in which the assignment has been doubtful. Some of these levels will also de-excite via electromagnetic emission. The comparison between the energy of proton that populate a state of {sup 12}C and the sum of the energies of the 3{alpha} emitted from the same state makes possible to determine the presence of electromagnetic disintegration ({gamma}) to lower states within {sup 12}C followed by the 3{alpha} break-up.

  7. ATP-dependent transport of bile acid intermediates across rat liver peroxisomal membranes.

    PubMed

    Une, Mizuho; Iguchi, Yusuke; Sakamoto, Tomoko; Tomita, Takashi; Suzuki, Yasuyuki; Morita, Masashi; Imanaka, Tsuneo

    2003-08-01

    The bile acid intermediate 3alpha,7alpha,12alpha-trihydroxy-5beta-cholestanoic acid (THCA) is converted to cholic acid exclusively in peroxisomes by the oxidative cleavage of the side chain. To investigate the mechanism by which the biosynthetic intermediates of bile acids are transported into peroxisomes, we incubated THCA or its CoA ester (THC-CoA) with isolated intact rat liver peroxisomes and analyzed their oxidation products, cholic acid and 3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-24-enoic acid. The oxidation of both THCA and THC-CoA was dependent on incubation time and peroxisomal proteins, and was stimulated by ATP. THC-CoA was efficiently oxidized to cholic acid and 3alpha,7alpha,12alpha-trihydroxy-5beta-cholest-24-enoic acid as compared with THCA, suggesting that THC-CoA is the preferred substrate for transport into peroxisomes. The oxidation of THC-CoA was significantly inhibited by sodium azide, verapamile, and N-ethylmaleimide. Furthermore, the stimulatory effect of ATP on the oxidation was not replaced by GTP or AMP. In addition, the ATP-dependent oxidation of THC-CoA was markedly inhibited by pretreatment of peroxisomes with proteinase K when peroxisomal matrix proteins were not degraded. These results suggest that an ATP-dependent transport system for THC-CoA exists on peroxisomal membranes.

  8. Absorption and transport of deuterium-substituted 2R,4'R,8'R-alpha-tocopherol in human lipoproteins

    SciTech Connect

    Traber, M.G.; Ingold, K.U.; Burton, G.W.; Kayden, H.J.

    1988-08-01

    Oral administration of a single dose of tri- or hexadeuterium substituted 2R,4'R,8'R-alpha-tocopheryl acetate (d3- or d6-alpha-T-Ac) to humans was used to follow the absorption and transport of vitamin E in plasma lipoproteins. Three hr after oral administration of d3-alpha-T-Ac (15 mg) to 2 subjects, plasma levels of d3-alpha-T were detectable; these increased up to 10 hr, reached a plateau at 24 hr, then decreased. Following administration of d6-alpha-T-Ac (15-16 mg) to 2 subjects, the percentage of deuterated tocopherol relative to the total tocopherol in chylomicrons increased more rapidly than the corresponding percentage in whole plasma. Chylomicrons and plasma lipoproteins were isolated from 2 additional subjects following administration of d3-alpha-T-Ac (140 or 60 mg). The percentage of deuterated tocopherol relative to the total tocopherol increased most rapidly in chylomicrons, then in very low density lipoproteins (VLDL), followed by essentially identical increases in low and high density lipoproteins (LDL and HDL, respectively) and lastly, in the red blood cells. This pattern of appearance of deuterated tocopherol is consistent with the concept that newly absorbed vitamin E is secreted by the intestine into chylomicrons; subsequently, chylomicron remnants are taken up by the liver from which the vitamin E is secreted in VLDL. The metabolism of VLDL in the circulation results in the simultaneous delivery of vitamin E into LDL and HDL.

  9. Distinct CDR3 Conformations in TCRs Determine the Level of Cross-Reactivity for Diverse Antigens, But Not the Docking Orientation

    SciTech Connect

    Jones, L.L.; Colf, L.A.; Stone, J.D.; Garcia, K.C.; Kranz, D.M.

    2009-05-18

    T cells are known to cross-react with diverse peptide MHC Ags through their {alpha}{beta} TCR. To explore the basis of such cross-reactivity, we examined the 2C TCR that recognizes two structurally distinct ligands, SIY-K{sup b} and alloantigen QL9-L{sup d}. In this study we characterized the cross-reactivity of several high-affinity 2C TCR variants that contained mutations only in the CDR3{alpha} loop. Two of the TCR lost their ability to cross-react with the reciprocal ligand (SIY-K{sup b}), whereas another TCR (m67) maintained reactivity with both ligands. Crystal structures of four of the TCRs in complex with QL9-L{sup d} showed that CDR1, CDR2, and CDR3{beta} conformations and docking orientations were remarkably similar. Although the CDR3{alpha} loop of TCR m67 conferred a 2000-fold higher affinity for SIY-K{sup b}, the TCR maintained the same docking angle on QL9-L{sup d} as the 2C TCR. Thus, CDR3{alpha} dictated the affinity and level of cross-reactivity, yet it did so without affecting the conserved docking orientation.

  10. Mechanochemical synthesis and crystal structure of alpha'-AlD3 and alpha-AlD3.

    PubMed

    Brinks, Hendrik W; Istad-Lem, Andreas; Hauback, Bjørn C

    2006-12-28

    AlD3 AlD3 was synthesized by ball milling of 3LiAlD4 + AlCl3. Planetary ball milling at room temperature resulted in a mixture of AlD3 (alpha and alpha') and Al in addition to LiCl, whereas cryomilling at 77 K resulted in only AlD3 and LiCl. The AlD3 obtained was a mixture of about 2/3alpha and 1/3alpha'. Alpha' was determined by powder neutron diffraction to take the beta-AlF3 structure with space group Cmcm and a = 6.470(3), b = 11.117(5), and c = 6.562(2) A. It is built up of corner-sharing AlD6 octahedra in an open structure with hexagonal holes of radius 3.9 A. Alpha' slowly decomposes during storage at 40 degrees C. Alpha-AlD3 is also described by a corner-sharing AlD6 network but in a more dense ReO3-type arrangement. Both AlD3 modifications have slightly shorter Al-D distances compared to Na3AlD6, Na2LiAlD6, and K2NaAlH6.

  11. Antioxidant activities and antitumor screening of extracts from cranberry fruit (Vaccinium macrocarpon).

    PubMed

    Yan, Xiaojun; Murphy, Brian T; Hammond, Gerald B; Vinson, Joe A; Neto, Catherine C

    2002-10-09

    Polyphenolic compounds in cranberries have been investigated to determine their role in protection against cardiovascular disease and some cancers. Extracts of whole fruit were assayed for radical-scavenging activity and tumor growth inhibition using seven tumor cell lines. Selective inhibition of K562 and HT-29 cells was observed from a methanolic extract in the range of 16-125 microg/mL. Radical-scavenging activity was greatest in an extract composed primarily of flavonol glycosides. Seven flavonol glycosides were isolated and purified from whole fruit for further evaluation; the anthocyanin cyanidin 3-galactoside was also purified for comparison with the flavonoids. Three flavonol monoglycosides were newly identified by (13)C NMR as myricetin 3-alpha-arabinofuranoside, quercetin 3-xyloside, and 3-methoxyquercetin 3-beta-galactoside (isorhamnetin); the other four isolated were the previously identified myricetin 3-beta-galactoside, quercetin 3-beta-galactoside, quercetin 3-alpha-arabinofuranoside, and quercetin 3-alpha-rhamnopyranoside. These compounds were evaluated for 1,1-diphenyl-2-picrylhydrazyl radical-scavenging activity and ability to inhibit low-density lipoprotein oxidation in vitro. Most of the flavonol glycosides showed antioxidant activity comparable or superior to that of vitamin E; cyanidin 3-galactoside showed activity superior to that of the flavonoids as well as vitamin E or Trolox in both antioxidant assays.

  12. AKT/SGK-sensitive phosphorylation of GSK3 in the regulation of L-selectin and perforin expression as well as activation induced cell death of T-lymphocytes

    SciTech Connect

    Bhavsar, Shefalee K.; Merches, Katja; Bobbala, Diwakar; Lang, Florian

    2012-08-17

    Highlights: Black-Right-Pointing-Pointer Akt/SGK dependent phosphorylation of GSK3{alpha},{beta} regulates T lymphocytes. Black-Right-Pointing-Pointer T cells from mice expressing Akt/SGK insensitive GSK3{alpha},{beta} (gsk3{sup KI}) release less IL-2. Black-Right-Pointing-Pointer CD4{sup +} cells from gsk3{sup KI} mice express less CD62L. Black-Right-Pointing-Pointer CD8{sup +} cells from gsk3{sup KI} mice are relatively resistant to activation induced cell death. Black-Right-Pointing-Pointer Perforin expression is enhanced in gsk3{sup KI} T cells. -- Abstract: Survival and function of T-lymphocytes critically depends on phosphoinositide (PI) 3 kinase. PI3 kinase signaling includes the PKB/Akt and SGK dependent phosphorylation and thus inhibition of glycogen synthase kinase GSK3{alpha},{beta}. Lithium, a known unspecific GSK3 inhibitor protects against experimental autoimmune encephalomyelitis. The present study explored, whether Akt/SGK-dependent regulation of GSK3 activity is a determinant of T cell survival and function. Experiments were performed in mutant mice in which Akt/SGK-dependent GSK3{alpha},{beta} inhibition was disrupted by replacement of the serine residue in the respective SGK/Akt-phosphorylation consensus sequence by alanine (gsk3{sup KI}). T cells from gsk3{sup KI} mice were compared to T cells from corresponding wild type mice (gsk3{sup WT}). As a result, in gsk3{sup KI} CD4{sup +} cells surface CD62L (L-selectin) was significantly less abundant than in gsk3{sup WT} CD4{sup +} cells. Upon activation in vitro T cells from gsk3{sup KI} mice reacted with enhanced perforin production and reduced activation induced cell death. Cytokine production was rather reduced in gsk3{sup KI} T cells, suggesting that GSK3 induces effector function in CD8{sup +} T cells. In conclusion, PKB/Akt and SGK sensitive phosphorylation of GSK3{alpha},{beta} is a potent regulator of perforin expression and activation induced cell death in T lymphocytes.

  13. Endogenous selective inhibitors of 11beta-hydroxysteroid dehydrogenase isoforms 1 and 2 of adrenal origin.

    PubMed

    Latif, Syed A; Pardo, Hector A; Hardy, Matthew P; Morris, David J

    2005-11-24

    In earlier studies [Latif, S.A., Sheff, M.F., Ribeiro, C.E., Morris, D.J., 1997. Selective inhibition of sheep kidney 11beta-hydroxysteroid-dehydrogenase isoform 2 activity by 5alpha-reduced (but not 5beta) derivatives of adrenocorticosteroids. Steroids 62, 230-237], only derivatives of steroid hormones possessing the 5alpha-Ring A-reduced configuration selectively inhibited 11beta-HSD2-dehydrogenase, whereas their 5beta-derivatives were inactive. This present study focuses on an expanded group of endogenous 11-oxygenated, 5alpha and 5beta-Ring A-reduced metabolites of adrenocorticosteroids, and progestogen and androgen steroid hormones. These substances were tested for their inhibitory properties against 11beta-HSD2, 11beta-HSD1-dehydrogenase and 11beta-HSD1 reductase. The present studies showed that the following compounds stand out as potent inhibitors. These are 5alpha-DH-corticosterone, 3alpha,5alpha-TH-corticosterone, 11beta-OH-progesterone, 11beta-OH-allopregnanolone, 11beta-OH-testosterone, and 11beta-OH-androstanediol, inhibitors of 11beta-HSD1-dehydrogenase; 3alpha,5alpha-TH-11-dehydro-corticosterone, 11-keto-progesterone, 11-keto-allopregnanolone, and 11-keto-3beta,5alpha-TH-testosterone, inhibitors of 11beta-HSD1 reductase; 3alpha,5alpha-TH-aldosterone, 5alpha-DH-corticosterone, 3alpha,5alpha-TH-corticosterone,11-dehydro-corticosterone, 3alpha,5alpha-TH-11-dehydro-corticosterone, 11beta-OH-progesterone, 11-keto-progesterone, 11beta-OH-allopregnanolone, 11-keto-allopregnanolone, 11beta-OH-testosterone, and 11-keto-testosterone, inhibitors of 11beta-HSD2. All of these substances have the potential to be derived from adrenally synthesized corticosteroids. Substances with similar structures to those described may help in the design of exogenous agents for the management of a variety of disease states involving 11beta-HSD isoenzymes.

  14. Metabolism of 4-hydroxyandrostenedione and 4-hydroxytestosterone: Mass spectrometric identification of urinary metabolites.

    PubMed

    Kohler, Maxie; Parr, Maria K; Opfermann, Georg; Thevis, Mario; Schlörer, Nils; Marner, Franz-Josef; Schänzer, Wilhelm

    2007-03-01

    4-Hydroxyandrost-4-ene-3,17-dione is a second generation, irreversible aromatase inhibitor and commonly used as anti breast cancer medication for postmenopausal women. 4-Hydroxytestosterone is advertised as anabolic steroid and does not have any therapeutic indication. Both substances are prohibited in sports by the World Anti-Doping Agency, and, due to a considerable increase of structurally related steroids with anabolic effects offered via the internet, the metabolism of two representative candidates was investigated. Excretion studies were conducted with oral applications of 100mg of 4-hydroxyandrostenedione or 200mg of 4-hydroxytestosterone to healthy male volunteers. Urine samples were analyzed for metabolic products using conventional gas chromatography-mass spectrometry approaches, and the identification of urinary metabolites was based on reference substances, which were synthesized and structurally characterized by nuclear magnetic resonance spectroscopy and high resolution/high accuracy mass spectrometry. Identified phase-I as well as phase-II metabolites were identical for both substances. Regarding phase-I metabolism 4-hydroxyandrostenedione (1) and its reduction products 3beta-hydroxy-5alpha-androstane-4,17-dione (2) and 3alpha-hydroxy-5beta-androstane-4,17-dione (3) were detected. Further reductive conversion led to all possible isomers of 3xi,4xi-dihydroxy-5xi-androstan-17-one (4, 6-11) except 3alpha,4alpha-dihydroxy-5beta-androstan-17-one (5). Out of the 17beta-hydroxylated analogs 4-hydroxytestosterone (18), 3beta,17beta-dihydroxy-5alpha-androstan-4-one (19), 3alpha,17beta-dihydroxy-5beta-androstan-4-one (20), 5alpha-androstane-3beta,4beta,17beta-triol (21), 5alpha-androstane-3alpha,4beta,17beta-triol (26) and 5alpha-androstane-3alpha,4alpha,17beta-triol (28) were identified in the post administration urine specimens. Furthermore 4-hydroxyandrosta-4,6-diene-3,17-dione (29) and 4-hydroxyandrosta-1,4-diene-3,17-dione (30) were determined as

  15. Monocyte chemoattractant protein-1 (MCP-1) inhibits the intestinal-like differentiation of monocytes.

    PubMed

    Spoettl, T; Hausmann, M; Herlyn, M; Gunckel, M; Dirmeier, A; Falk, W; Herfarth, H; Schoelmerich, J; Rogler, G

    2006-07-01

    Monocytes (MO) migrating into normal, non-inflamed intestinal mucosa undergo a specific differentiation resulting in a non-reactive, tolerogenic intestinal macrophage (IMAC). Recently we demonstrated the differentiation of MO into an intestinal-like macrophage (MAC) phenotype in vitro in a three-dimensional cell culture model (multi-cellular spheroid or MCS model). In the mucosa of patients with inflammatory bowel disease (IBD) in addition to normal IMAC, a reactive MAC population as well as increased levels of monocyte chemoattractant protein 1 (MCP-1) is found. The aim of this study was to investigate the influence of MCP-1 on the differentiation of MO into IMAC. MCS were generated from adenovirally transfected HT-29 cells overexpressing MCP-1, macrophage inflammatory protein 3 alpha (MIP-3alpha) or non-transfected controls and co-cultured with freshly elutriated blood MO. After 7 days of co-culture MCS were harvested, and expression of the surface antigens CD33 and CD14 as well as the intracellular MAC marker CD68 was determined by flow-cytometry or immunohistochemistry. MCP-1 and MIP-3alpha expression by HT-29 cells in the MCS was increased by transfection at the time of MCS formation. In contrast to MIP-3alpha, MCP-1 overexpression induced a massive migration of MO into the three-dimensional aggregates. Differentiation of IMAC was disturbed in MCP-1-transfected MCS compared to experiments with non-transfected control aggregates, or the MIP-3alpha-transfected MCS, as indicated by high CD14 expression of MO/IMAC cultured inside the MCP-1-transfected MCS, as shown by immunohistochemistry and FACS analysis. Neutralization of MCP-1 was followed by an almost complete absence of monocyte migration into the MCS. MCP-1 induced migration of MO into three-dimensional spheroids generated from HT-29 cells and inhibited intestinal-like differentiation of blood MO into IMAC. It may be speculated that MCP-1 could play a role in the disturbed IMAC differentiation in IBD mucosa.

  16. Defective peroxisomal cleavage of the C27-steroid side chain in the cerebro-hepato-renal syndrome of Zellweger.

    PubMed Central

    Kase, B F; Björkhem, I; Hågå, P; Pedersen, J I

    1985-01-01

    Based on in vitro work with rat liver, we recently suggested that the peroxisomal fraction is most important for the oxidation of 3 alpha, 7 alpha, 12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) into cholic acid. The cerebro-hepato-renal syndrome of Zellweger is a fatal recessive autosomal disorder, the most characteristic histological feature of which is a virtual absence of peroxisomes in liver and kidneys. This disease offers a unique opportunity to evaluate the relative importance of peroxisomes in bile acid biosynthesis. A child with Zellweger syndrome was studied in the present work. In accordance with previous work, there was a considerable accumulation of THCA, 3 alpha, 7 alpha, 12 alpha, 24-tetrahydroxy-5 beta-cholestanoic acid (24-OH-THCA), 3 alpha, 7 alpha, 12 alpha-trihydroxy-27-carboxymethyl-5 beta-cholestan-26-oic acid (C29-dicarboxylic acid), and 3 alpha, 7 alpha-dihydroxy-5 beta-cholestanoic acid in serum. In addition, a tetrahydroxylated 5 beta-cholestanoic acid with all the hydroxyl groups in the steroid nucleus was found. 3H-Labeled 5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol was administered intravenously together with 14C-labeled cholic acid. There was a rapid incorporation of 3H in THCA and a slow incorporation into cholic acid. The specific radioactivity of 3H in THCA was about one magnitude higher than that in cholic acid. The conversion was evaluated by following the increasing ratio between 3H and 14C in biliary cholic acid. The rate of incorporation of 3H in cholic acid was considerably less than previously reported in experiments with healthy subjects, and the maximal conversion of the triol into cholic acid was only 15-20%. About the same rate of conversion was found after oral administration of 3H-THCA. Both in the experiment with 3H-5 beta-cholestane-3 alpha, 7 alpha, 12 alpha-triol and with 3H-THCA, there was an efficient incorporation of 3H in the above unidentified tetrahydroxylated 5 beta-cholestanoic acid. There was

  17. The oestrogenic effects of gestodene, a potent contraceptive progestin, are mediated by its A-ring reduced metabolites.

    PubMed

    Lemus, A E; Zaga, V; Santillán, R; García, G A; Grillasca, I; Damián-Matsumura, P; Jackson, K J; Cooney, A J; Larrea, F; Pérez-Palacios, G

    2000-06-01

    Gestodene (17 alpha-ethynyl-13 beta-ethyl-17 beta-hydroxy-4, 15-gonadien-3-one) is the most potent synthetic progestin currently available and it is widely used as a fertility regulating agent in a number of contraceptive formulations because of its high effectiveness, safety and acceptability. The observation that contraceptive synthetic progestins exert hormone-like effects other than their progestational activities, prompted us to investigate whether gestodene (GSD) administration may induce oestrogenic effects, even though the GSD molecule does not interact with intracellular oestrogen receptors (ER). To assess whether GSD may exert oestrogenic effects through some of its neutral metabolites, a series of experimental studies were undertaken using GSD and three of its A-ring reduced metabolites. Receptor binding studies by displacement analysis confirmed that indeed GSD does not bind to the ER, whereas its 3 beta,5 alpha-tetrahydro reduced derivative (3 beta GSD) interacts with a relative high affinity with the ER. The 3 alpha,5 alpha GSD isomer (3 alpha GSD) also binds to the ER, though to a lesser extent. The ability of the A-ring reduced GSD derivatives to induce oestrogenic actions was evaluated by the use of two different molecular bioassays: (a) transactivation of a yeast system co-transfected with the human ER alpha (hER alpha) gene and oestrogen responsive elements fused to the beta-galactosidase reporter vector and (b) transactivation of the hER alpha-mediated transcription of the chloramphenicol acetyl transferase (CAT) reporter gene in a HeLa cells expression system. The oestrogenic potency of 3 beta GSD was also assessed by its capability to induce oestrogen-dependent progestin receptors (PR) in the anterior pituitary of castrated female rats. The results demonstrated that 3 beta GSD and 3 alpha GSD were able to activate, in a dose-dependent manner, the hER alpha-mediated transcription of both the beta-galactosidase and the CAT reporter genes in the

  18. The first biantennary bacterial secondary cell wall polymer and its influence on S-layer glycoprotein assembly.

    PubMed Central

    Steindl, Christian; Schäffer, Christina; Wugeditsch, Thomas; Graninger, Michael; Matecko, Irena; Müller, Norbert; Messner, Paul

    2002-01-01

    The cell surface of Aneurinibacillus thermoaerophilus DSM 10155 is covered with a square surface (S)-layer glycoprotein lattice. This S-layer glycoprotein, which was extracted with aqueous buffers after a freeze-thaw cycle of the bacterial cells, is the only completely water-soluble S-layer glycoprotein to be reported to date. The purified S-layer glycoprotein preparation had an overall carbohydrate content of 19%. Detailed chemical investigations indicated that the S-layer O-glycans of previously established structure accounted for 13% of total glycosylation. The remainder could be attributed to a peptidoglycan-associated secondary cell wall polymer. Structure analysis was performed using purified secondary cell wall polymer-peptidoglycan complexes. NMR spectroscopy revealed the first biantennary secondary cell wall polymer from the domain Bacteria, with the structure alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->4)-[alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->4)-beta-L-Gal p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)]-beta-L-Man p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-beta-L-Man p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->3)-alpha-L-Glc p NAc-(1-->O)-PO(2)(-)-O-PO(2)(-)-(O-->6)-MurNAc- (where MurNAc is N -acetylmuramic acid). The neutral polysaccharide is linked via a pyrophosphate bond to the C-6 atom of every fourth N -acetylmuramic acid residue, in average, of the A1gamma-type peptidoglycan. In vivo, the biantennary polymer anchored the S-layer glycoprotein very effectively to the cell wall, probably due to the doubling of motifs for a proposed lectin-like binding between the polymer and the N-terminus of the S-layer protein. When the cellular support was removed during S-layer glycoprotein isolation, the co-purified polymer mediated the solubility of the S

  19. Riemannian geometry of thermodynamics and systems with repulsive power-law interactions.

    PubMed

    Ruppeiner, George

    2005-07-01

    A Riemannian geometric theory of thermodynamics based on the postulate that the curvature scalar R is proportional to the inverse free energy density is used to investigate three-dimensional fluid systems of identical classical point particles interacting with each other via a power-law potential energy gamma r(-alpha) . Such systems are useful in modeling melting transitions. The limit alpha-->infinity corresponds to the hard sphere gas. A thermodynamic limit exists only for short-range (alpha>3) and repulsive (gamma>0) interactions. The geometric theory solutions for given alpha>3 , gamma>0 , and any constant temperature T have the following properties: (1) the thermodynamics follows from a single function b (rho T(-3/alpha) ) , where rho is the density; (2) all solutions are equivalent up to a single scaling constant for rho T(-3/alpha) , related to gamma via the virial theorem; (3) at low density, solutions correspond to the ideal gas; (4) at high density there are solutions with pressure and energy depending on density as expected from solid state physics, though not with a Dulong-Petit heat capacity limit; (5) for 3<alpha<3.7913 , the solution goes from the low to the expected high density limit smoothly; (6) for alpha>3.7913 a phase transition is required to go between these regimes; (7) for any alpha>3 we may include a first-order phase transition, which is expected from computer simulations; and (8) if alpha-->infinity, the density approaches a finite value as the pressure increases to infinity, with the pressure diverging logarithmically in the density difference.

  20. Synthesis of deuterium-labeled 17-hydroxyprogesterone suitable as an internal standard for isotope dilution mass spectrometry

    SciTech Connect

    Shimizu, K.; Yamaga, N.; Kohara, H.

    1988-03-01

    A synthesis is reported of 17-hydroxyprogesterone, labeled with four atoms of deuterium at ring C and suitable for use as an internal standard for isotope dilution mass spectrometry. Base-catalyzed equilibration of methyl 3 alpha-acetoxy-12-oxo-cholanate (III) with /sup 2/H/sub 2/O, followed by reduction of the 12-oxo group by the modified Wolff-Kisher method using (/sup 2/H)diethylene glycol and (/sup 2/H)hydrazine hydrate afforded (11,11,12,12,23,23(-2)H)lithocholic acid (V). The Meystre-Miescher degradation of the side chain of V yielded 3 alpha-hydroxy-5 beta-(11,11,12,12(-2)H)pregnan-20-one (X). Oxidation of the 3,20-enol-diacetate of X with perbenzoic acid followed by saponification afforded 3 alpha,17-dihydroxy-5 beta-(11,11,12,12(-2)H)pregnan-20-one (XI). Oxidation of XI with N-bromoacetamide yielded 17-hydroxy-5 beta-(11,11,12,12(-2)H)pregnane-3,20-dione (XII). Bromination of XII followed by dehydrobromination yielded 17-hydroxy-(11,11,12,12(-2)H) progesterone (XIV), consisting of 0.3% /sup 2/H0-, 1.1% /sup 2/H/sub 1/-, 8.6% /sup 2/H/sub 2/-, 37.1% /sup 2/H/sub 3/-, 52.1% /sup 2/H/sub 4/-, and 0.8% /sup 2/H/sub 5/-species.

  1. Increased 21-hydroxylase and shutdown of C(17,20) lyase activities in testicular tissues of the grouper (Epinephelus coioides) during 17alpha-methyltestosterone-induced sex inversion.

    PubMed

    Lee, S T; Lam, T J; Tan, C H

    2002-05-01

    The metabolism in vitro of [(3)H]17-hydroxyprogesterone by gonadal tissues of the grouper (Epinephelus coioides) during 17alpha-methyltestosterone (MT)-induced female-to-male sex inversion was examined. In the female phase, C(17,20) lyase, 5beta-reductase, 3alpha/beta-HSD, 20beta-HSD, and 17beta-HSD activities resulted in the biosynthesis of 5beta-pregnans and 5beta-androstanes (including 5beta-androstane-3alpha/beta, 17beta-diol, 3alpha/beta, 17alpha-dihydroxy-5beta-pregnen-20-one, and 5beta-androstane-3,17-dione). In the MT-induced male phase, however, the abrogation of C(17,20) lyase activity and the concomitant activation of 21alpha-hydroxylase/11beta-hydroxylase resulted in the preferential synthesis of polar 21alpha-hydroxlyated 5beta-pregnans (5beta-pregnan-3beta,17alpha,20beta,21alpha-tetrol and 3beta,20beta,21alpha-trihydroxy-5beta-pregnan-3-one) and corticosteroids (11-deoxycortisol and cortisol). Interestingly, synthesis of these 21alpha-hydroxylated 5beta-pregnans and corticosteroids was uniquely compartmentalized in only testicular tissues of the MT-induced males. This study shows that there is selective activation of specific steroidogenic enzymes in the different sexual phases leading to the synthesis of metabolites that may be involved in regulating sex inversion of the grouper. (c) 2002 Elsevier Science (USA).

  2. A novel rabconnectin-3-binding protein that directly binds a GDP/GTP exchange protein for Rab3A small G protein implicated in Ca(2+)-dependent exocytosis of neurotransmitter.

    PubMed

    Kawabe, Hiroshi; Sakisaka, Toshiaki; Yasumi, Masato; Shingai, Tatsushi; Izumi, Genkichi; Nagano, Fumiko; Deguchi-Tawarada, Maki; Takeuchi, Masakazu; Nakanishi, Hiroyuki; Takai, Yoshimi

    2003-06-01

    Rab3A, a member of the Rab3 small G protein family, regulates Ca2+-dependent exocytosis of neurotransmitter. The cyclical activation and inactivation of Rab3A are essential for the Rab3A action in exocytosis. GDP-Rab3A is activated to GTP-Rab3A by Rab3 GDP/GTP exchange protein (Rab3 GEP) and GTP-Rab3A is inactivated to GDP-Rab3A by Rab3 GTPase-activating protein (Rab3 GAP). We have recently found a novel protein, named rabconnectin-3, which is co-immunoprecipitated with Rab3 GEP or GAP from the extract of the crude synaptic vesicle (CSV) fraction of rat brain. Rabconnectin-3 is abundantly expressed in the brain where it is associated with synaptic vesicles. We have found that two more proteins are co-immunoprecipitated with Rab3 GEP from the CSV fraction of rat brain. We attempted here to isolate and characterize one of them. We determined its partial amino acid sequence, cloned its cDNA from a human cDNA library, and determined its primary structure. The protein consisted of 1490 amino acids (aa) and showed a calculated molecular weight of 163808. The protein had 7 WD domains. The protein was abundantly expressed in the brain where it co-localized with rabconnectin-3 on synaptic vesicles. The protein formed a stable complex with rabconnectin-3. We named this protein rabconnectin-3beta and renamed rabconnectin-3 rabconnectin-3alpha. Rabconnectin-3beta, but not rabconnectin-3alpha, directly bound Rab3 GEP. Neither rabconnectin-3alpha nor -3beta directly bound Rab3 GAP. These results indicate that rabconnectin-3 consists of the alpha and beta subunits and binds directly Rab3 GEP through the beta subunit and indirectly Rab3 GAP through an unidentified molecule(s).

  3. Metabolism of C26 bile alcohols in the bullfrog, Rana catesbeiana

    SciTech Connect

    Noma, Y.; Kihira, K.; Kuramoto, T.; Hoshita, T.

    1988-03-01

    Metabolism of C26 bile alcohols in the bullfrog, Rana catesbeiana, was studied. (24-14C)-24-Dehydro-26-deoxy-5 beta-ranol (3 alpha,7 alpha,12 alpha-trihydroxy-27-nor-5 beta-cholestan-24-one) was chemically synthesized from (24-14C)cholic acid and incubated with bullfrog liver homogenate fortified with NADPH. 24-Dehydro-26-deoxy-5 beta-ranol was shown to be converted into both 26-deoxy-5 beta-ranol and 24-epi-26-deoxy-5 beta-ranol ((24S)- and (24R)-27-nor-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24-tetrols) in addition to 5 beta-ranol ((24R)-27-nor-5 beta-cholestane-3 alpha,7 alpha,12 alpha,24,26-pentol), which is the major bile alcohol of the bullfrog. (24-3H)-26-Deoxy-5 beta-ranol and (24-3H)-24-epi-26-deoxy-5 beta-ranol were prepared from 24-dehydro-26-deoxy-5 beta-ranol by reduction with sodium (3H) borohydride and administered respectively to two each of four bullfrogs by intraperitoneal injection. After 24 h, labeled 5 beta-ranol was isolated from the bile of the bullfrogs that received (24-3H)-26-deoxy-5 beta-ranol. In contrast little if any radioactivity could be detected in 5 beta-ranol or its 24-epimer after administration of (24-3H)-24-epi-26-deoxy-5 beta-ranol.

  4. Comparative studies on the constituents of ophiopogonis tuber and its congeners. VIII. Studies on the glycosides of the subterranean part of Ophiopogon japonicus Ker-Gawler cv. Nanus.

    PubMed

    Asano, T; Murayama, T; Hirai, Y; Shoji, J

    1993-03-01

    Two monoterpene glycosides, tentatively named OJV-I (1) and OJV-II (2), and eight steroidal glycosides, tentatively named OJV-III (3), OJV-IV (4), OJV-V (5), OJV-VI (6), OJV-VII (7), OJV-VIII (8), OJV-IX (9) and OJV-X (10), were isolated from the butanol-soluble fraction of the fresh subterranean part of Ophiopogon japonicus KER-GAWLER cv. Nanus. Among these compounds, 1, 2, 3, 4, 5, 6 and 7 were identified as l-borneo1 O-beta-D-glucopyranoside, l-borneo1 O-beta-D-apiofuranosyl (1-->6)-beta-D-glucopyranoside, ophiopogonin B, glycoside C, ophiopogonin D, Ls-10, and ruscogenin 1-O-sulfate, respectively. The structures of compounds 8, 9, and 10 were established to be (23S,24S,25S)-23,24-dihydroxyruscogenin 1-O-[ alpha-L-rhamnopyranosyl(1-->2)] [beta-D-xylopyranosyl(1-->3)]-alpha-L-arabinopyranoside 24-O-beta-D-fucopyranoside, (23S,24S,25S)-23,24-dihydroxyruscogenin I-O-[alpha-L-2,3,4-tri-O-acetylrhamnopyranosyl(1-->2)][beta-D-xylo pyranosyl(1-->3)]-alpha-L-arabinopyranoside 24-O-beta-D-fucopyranoside, and (23S,24S,25S)-23,24-dihydroxyruscogenin 1-O-[alpha-L-2,3,4-tri-O-acetylrhamnopyranosyl(1-->2)] [beta-D-xylopyranosyl(1-->3)]=alpha-L-arabinopyranoside 24-O-beta-D-fucopyranoside, respectively.

  5. [Measurement of the transport activities of bile salt export pump using chemiluminescence detection method].

    PubMed

    Yamaguchi, Kana; Murai, Tsuyoshi; Yabuuchi, Hikaru; Hui, Shu-Ping; Kurosawa, Takao

    2010-05-01

    Monovalent bile acids, such as taurine- and glycine-conjugated bile acids, are excreted into bile by bile salt export pumps (BSEP, ABCB11). Human BSEP (hBSEP) is physiologically important because it was identified as the gene responsible for the genetic disease: progressive familial intrahepatic cholestasis type 2 (PFIC-2). The evaluation of the inhibitory effect of hBSEP transport activity provides significant information for predicting toxic potential in the early phase of drug development. The role and function of hBSEP have been investigated by the examination of the ATP-dependent transport of radioactive isotopically (RI)-labeled bile acid such as a tritium labeled taurocholic acid, in membrane vesicles obtained from hBSEP-expressing cells. The chemiluminescence detection method using 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD) had been developed for a simple analysis of bile acids in human biological fluids. This method is extremely sensitive and it may be applicable for the measurements of bile acid transport activities by hBSEP vesicles without using RI-labeled bile acid. The present paper deals with an application of the chemiluminescence detection method using 3alpha-HSD with enzyme cycling method to the measurement of ATP-dependent transport activities of taurocholic acid (T-CA) in membrane vesicles obtained from hBSEP-expressing Sf9 cells. Calibration curves for T-CA was linear over the range from 10 to 400 pmol/ml. The values of the kinetic parameters for hBSEP vesicles obtained by the chemiluminescence detection method were comparable with the values of that obtained by liquid chromatography-mass spectrometry method. This assay method was highly useful for the measurements of bile acid transport activities.

  6. Skin of the male African catfish, Clarias gariepinus: a source of steroid glucuronides

    SciTech Connect

    Ali, S.A.; Schoonen, W.G.; Lambert, J.G.; Van den Hurk, R.; Van Oordt, P.G.

    1987-06-01

    Steroid metabolism in the skin of mature male African catfish, Clarias gariepinus, reared in the laboratory, was studied in vitro by tissue incubations with (/sup 3/H)pregnenolone, (/sup 3/H)dehydroepiandrosterone, (/sup 3/H)17 alpha-hydroxyprogesterone, (/sup 3/H)androstenedione, (/sup 14/C)11 beta-hydroxyandrostenedione, and (/sup 3/H)testosterone as precursors. While pregnenolone was not converted to any other steroid, dehydroepiandrosterone was transformed mainly to 5-androstene-3 beta, 17 beta-diol. The products of 17 alpha-hydroxyprogesterone incubations were 5 beta-pregnane-3 alpha,17 alpha-diol-20-one, 5 beta-pregnane-3 alpha,17 alpha, 20 beta-triol, and 5 beta-pregnan-17 alpha-o1-3,20-dione. The major steroids of androstenedione incubations were etiocholanolone, testosterone, and androsterone. Testosterone was converted mainly to etiocholanolone and androstenedione, and only small quantities of 11 beta-hydroxytestosterone, 11-ketotestosterone, and 11-ketoandrostenedione were the metabolites found in 11 beta-hydroxyandrostenedione incubation. These results demonstrated the presence of the enzymes 5 alpha- and 5 beta-reductases and 3 alpha-, 11 beta-, 17 beta-, and 20 beta-hydroxysteroid dehydrogenases in the skin. From enzymehistochemical results it appeared that the steroid conversions take place in the epithelial cells. Moreover, the presence of UDP-glucose dehydrogenase, an enzyme involved in the synthesis of glucuronic acid, in these cells indicates the possibility of steroid glucuronide formation. Indeed significant amounts of water-soluble steroid conjugates, particularly 5 beta-dihydrotestosterone- and testosterone-glucuronide, were found in the incubations with androstenedione and testosterone, indicating the presence of the UDP-glucuronosyl transferase in the catfish skin.

  7. Anti-inflammatory ergostanes from the basidiomata of Antrodia salmonea.

    PubMed

    Shen, Chien-Chang; Wang, Yea-Hwey; Chang, Tun-Tschu; Lin, Lie-Chwen; Don, Ming-Jaw; Hou, Yu-Chang; Liou, Kuo-Tong; Chang, Shiou; Wang, Wen-Yen; Ko, Han-Chieh; Shen, Yuh-Chiang

    2007-09-01

    Three new anti-oxidative ergostanes, methyl antcinate L (1), antcin M (2), and methyl antcinate K (3), together with nine additional known compounds, 3-ketodehydrosulphurenic acid, sulphurenic acid, dehydrosulphurenic acid, 3beta,15alpha-dihydroxylanosta-7,9(11),24-trien-21-oic acid, zhankuic acid A, zhankuic acid B, zhankuic acid C, antcin C, and antcin K were isolated from the basidiomata of Antrodia salmonea, a newly identified species of Antrodia (Polyporaceae) in Taiwan. These three new compounds were identified as methyl 3alpha,7alpha,12alpha-trihydroxy-4alpha-methylergosta-8,24(29)-dien-11-on-26-oate (1), 3alpha,12alpha-dihydroxy-4alpha-methylergosta-8,24(29)-dien-11-on-26-oic acid (2), and methyl 3alpha,4beta,7beta-trihydroxy-4alpha-methylergosta-8,24(29)-dien-11-on-26-oate (3) by spectroscopic analysis. We studied their antioxidative potential on the production of reactive oxygen species and nitric oxide (NO) in neutrophils and microglial cells, respectively. Compounds 1-3 displayed potent antioxidative activity with IC50 values of around 2.0-8.8 microM that was partially due to inhibition (6-67%) of NADPH oxidase activity but not through direct radical-scavenging properties. Compounds 1-3 also inhibited NO production with IC50 values of around 1.7-16.5 microM and were more potent than a non-specific NOS inhibitor. We conclude that these three new compounds 1, 2, and 3 exhibit anti-inflammatory activities in activated inflammatory cells.

  8. Definition of the surface antigens of Mycobacterium malmoense and use in studying the etiology of a form of mycobacteriosis.

    PubMed Central

    McNeil, M; Tsang, A Y; McClatchy, J K; Stewart, C; Jardine, I; Brennan, P J

    1987-01-01

    Mycobacterium malmoense is the latest of a roster of atypical mycobacteria implicated in pulmonary infections. Yet it lacks recognizable phenotypic features to allow its ready identification. Some 23 clinical isolates of M. malmoense were examined for homologous seroagglutination reactions and characteristic surface antigens. One group showed concordant agglutination interreactions and an identical spectrum of glycolipids and are regarded as M. malmoense sensu stricto. The glycolipids are of the newly found, trehalose-containing lipooligosaccharide class. De-O-acylation followed by high-pressure liquid chromatography revealed one major and several minor oligosaccharides. Partial acidic cleavage to release glycosidically linked trehalose, alpha-mannosidase digestion to demonstrate the presence of a non-reducing-end mannobiose, perdeuteriomethylation, partial acid hydrolysis, reduction, and O ethylation, combined with 1H nuclear magnetic resonance and electron impact and fast-atom bombardment mass spectrometry revealed the structure of the major oligosaccharide as alpha-D-Manp-(1----3) -alpha-D-Manp-(1----[2-alpha-L-Rhap-(1--]4--3)-alpha-L-Rh ap- (1----3)-alpha-D-Glcp-(1----1)-alpha-D-Glcp, in which two of the 2-alpha-L-Rhap residues are O methylated at C-3. (Man, mannose; Rha, rhamnose; Glc, glucose; p, pyranosyl). The structures of the minor oligosaccharides were also determined; they differ at the distal nonreducing end. The dominant oligosaccharide was acylated by octanoate, 2-methyleicosanoate, and 2,4-dimethylpentacosanoate to yield the major species-specific surface antigen of M. malmoense, which we regard as the most characteristic feature of the pathogen. Images PMID:3597323

  9. Distinct epitopes for anti-glomerular basement membrane alport alloantibodies and goodpasture autoantibodies within the noncollagenous domain of alpha3(IV) collagen: a janus-faced antigen.

    PubMed

    Wang, Xu-Ping; Fogo, Agnes B; Colon, Selene; Giannico, Giovanna; Abul-Ezz, Sameh R; Miner, Jeffrey H; Borza, Dorin-Bogdan

    2005-12-01

    Alport posttransplantation anti-glomerular basement membrane (GBM) nephritis is mediated by alloantibodies against the noncollagenous (NC1) domains of the alpha3alpha4alpha5(IV) collagen network, which is present in the GBM of the allograft but absent from Alport kidneys. The specificity of kidney-bound anti-GBM alloantibodies from a patient who had autosomal recessive Alport syndrome (ARAS) and developed posttransplantation nephritis was compared with that of Goodpasture autoantibodies from patients with autoimmune anti-GBM disease. Allograft-eluted alloantibodies reacted specifically with alpha3alpha4alpha5 NC1 hexamers, targeting their alpha3NC1 and alpha4NC1 subunits, and recognized a noncontiguous alloepitope formed jointly by the E(A) and E(B) regions of alpha3NC1 domain. In contrast, human Goodpasture autoantibodies recognized the separate E(A) and E(B) autoepitopes of alpha3NC1 but not the composite alloepitope. Molecular modeling of alpha3NC1 revealed that the alloepitope is more accessible within the NC1 hexamers than the partially sequestered Goodpasture autoepitopes. Overall, the specificity of alloantibodies indicated a selective lack of immune tolerance toward the alpha3 and alpha4(IV) collagen chains not expressed in patients with ARAS. Using COL4A3 knockout mice, a model of ARAS, it was shown further that acid-dissociated rather than native alpha3alpha4alpha5 NC1 hexamers elicited murine anti-GBM antibodies most closely resembling human ARAS alloantibodies. In contrast, alpha3NC1 monomers elicited Goodpasture-like murine antibodies, targeting the E(A) and E(B) autoepitopes. Thus, the identity of alpha3NC1 epitopes targeted by anti-GBM antibodies is strongly influenced by the molecular organization of the immunogen. These findings suggest that different isoforms of alpha3(IV) collagen may be implicated in the pathogenesis of ARAS posttransplantation anti-GBM nephritis and Goodpasture disease.

  10. Progesterone metabolism by the hypothalamus, pituitary, and uterus of the rat during pregnancy

    SciTech Connect

    Marrone, B.L.; Karavolas, H.J.

    1981-07-01

    Metabolites of (/sup 3/H)progesterone were quantitated from incubations of hypothalamus, pituitary, and uterus of rats during different stages of pregnancy. The hypothalamus, anterior pituitary, and a section of uterus from five rats on Days 1, 8, 15, and 21 of pregnancy were incubated individually with (3H)progesterone and analyzed for metabolite formation by reverse isotopic dilution analysis. The radioactive metabolites present were 5 alpha-pregnane-3,20-dione (5 alpha-DHP), 3 alpha-hydroxy-5 alpha-pregnan-20-one, 20 alpha-hydroxy-4-pregnen-3-one, 20 alpha-hydroxy-5 alpha-pregnan-3-one, and 5 alpha-pregnane-3 alpha, 20 alpha-diol. The major metabolite formed by the hypothalamus and pituitary was 5 alpha-DHP. In the pituitary samples, formation of 5 alpha-DHP was decreased on Days 15 and 21 of pregnancy compared to Day 1, and formation of 20 alpha-hydroxy-5 alpha-pregnan-3-one was decreased on Day 21 compared to Day 1. In the uterine samples, 3 alpha-hydroxy-5 alpha-pregnan-20-one was the major metabolite formed at all stages of pregnancy. The formation of all metabolic products of progesterone by the uterus was increased on Day 21 compared to Days 1, 8, and 15 of pregnancy. No changes in the formation of progesterone metabolites were observed in the hypothalamic samples during pregnancy. It is concluded that there are different profiles in the in vitro metabolism of (3H)progesterone by the hypothalamus, pituitary, and uterus of the rat during the course of pregnancy.

  11. Sexuality and fertility in women with Addison's disease.

    PubMed

    Erichsen, Martina M; Husebye, Eystein S; Michelsen, Trond M; Dahl, Alv A; Løvås, Kristian

    2010-09-01

    Females with primary adrenal insufficiency (Addison's disease) have reduced levels of circulating androgens, which are allegedly important for sexual functioning. The aim was to determine peripheral androgen status, sexual functioning, and birth rates in Addison's disease females. In a postal survey, all 269 females in the Norwegian Addison's registry were invited to complete the Sexual Activity Questionnaire (SAQ) and registration of childbirths. Blood samples were analyzed for 5alpha-androstane-3alpha,17beta-diol-3-glucuronide (3alpha-Diol-G) and compared with blood donor levels. The SAQ scores were compared with 740 age-matched controls from the general population and 234 women subjected to risk-reducing salpingo-oophorectomy. Fertility was estimated as standardized incidence ratio for birth; the expected number of births was estimated from population statistics. The SAQ was completed by 174 (65%) of the Addison's patients. Those not taking DHEA had significantly lower 3alpha-Diol-G levels than blood donors (mean, 0.53 vs. 2.2 ng/ml; P < 0.0001), whereas those on DHEA treatment had elevated levels (mean, 5.8 vs. 2.2 ng/ml; P = 0.002). The Addison's disease females were equally sexually active as the controls, but they reported significantly higher pleasure and less discomfort. They reported lower pleasure but less discomfort than the risk-reducing salpingo-oophorectomy women. The fertility was significantly reduced in females with Addison's disease; 54 children were born to mothers with established diagnosis (87.5 expected), yielding a standardized incidence ratio for birth of 0.69 (confidence interval, 0.52-0.86). Despite androgen depletion, females with Addison's disease do not report impaired sexuality. The fertility is reduced after the diagnosis is made; the reasons for this remain unknown.

  12. KSC-2011-1799

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA’s Kennedy Space Center in Florida, workers accompany space shuttle Endeavour as it is backed out of Orbiter Processing Facility-2 for its move, or "rollover," to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Frankie Martin

  13. KSC-2011-1788

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA’s Kennedy Space Center in Florida, workers accompany space shuttle Endeavour as it is backed out of Orbiter Processing Facility-2 for its move, "or rollover," to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  14. KSC-2011-1787

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- In Orbiter Processing Facility-2 at NASA’s Kennedy Space Center in Florida, workers prepare space shuttle Endeavour for its move, or "rollover," from its hangar to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  15. KSC-2011-1785

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- In Orbiter Processing Facility-2 at NASA’s Kennedy Space Center in Florida, workers prepare space shuttle Endeavour for its move, or "rollover," from its hangar to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  16. KSC-2011-1790

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, employees hold up a banner to commemorate space shuttle Endeavour's STS-134 mission as it is transported from Orbiter Processing Facility-2 to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  17. KSC-2011-1792

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, workers accompany space shuttle Endeavour as it is being transported from Orbiter Processing Facility-2 to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  18. KSC-2011-1791

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, workers accompany space shuttle Endeavour as it is being transported from Orbiter Processing Facility-2 to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  19. KSC-2011-1786

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- In Orbiter Processing Facility-2 at NASA’s Kennedy Space Center in Florida, workers prepare space shuttle Endeavour for its move, or "rollover," from its hangar to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  20. KSC-2011-1800

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, employees hold up a banner to commemorate space shuttle Endeavour's STS-134 mission as it is transported from Orbiter Processing Facility-2 to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Frankie Martin

  1. Bent three-{alpha} linear-chain structure of {sup 13}C

    SciTech Connect

    Furutachi, N.; Kimura, M.

    2011-02-15

    The stability of the three-{alpha} linear-chain structure of {sup 13}C has been investigated with a microscopic 3{alpha}+n model. We have found two excited rotational bands that have developed a three-{alpha} cluster structure in {sup 13}C. The lower band built on 3/2{sub 2}{sup -} state at 11.4 MeV has the bent three-{alpha} linear-chain structure, and this structure is stable against the bending motion of three-{alpha} clusters.

  2. Large refractive nonlinearities and two-photon absorption in aryl-substituted dithiolenes

    NASA Astrophysics Data System (ADS)

    Kershaw, Stephen V.; Oliver, Stephen N.; Manning, Robert J.; Rush, James D.; Hill, Callum A. S.; Underhill, Allan E.; Charlton, Adam

    1993-12-01

    We have investigated some aryl substituted dithiolenes which show enhanced nonlinearity and their incorporation into polymer films. These neutral molecules show strong near infra-red absorption whose position is determined primarily by the extent of electron delocalization and the metal atom to which the sulphur ligands are attached. The presence of electron donating groups bathochromically shifts the band and selective detuning from the band is possible to arrive at optimum values for the important ration of nonlinear susceptibility to linear absorption, (chi) (3)/(alpha) . Nonlinear optical characterization of these materials by degenerate four wave mixing and z-scan is described.

  3. [Study on the chemical constituents in Pouzolzia zeylanica].

    PubMed

    Fu, Ming; Niu, You-Ya; Yu, Juan; Kong, Qing-Tong

    2012-11-01

    To study the chemical constituents of Pouzolzia zeylanica. Many chromatography means were used in separation and purification, and the structures of all compounds were identified by the means of spectroscopic analysis and physicochemical properties. 14 compounds were elucidated as: beta-sitosterol (1), daucosterol (2), oleanolic acid (3), epicatechin (4), alpha-amyrin (5), eugenyl-beta-rutinoside (6), 2alpha, 3alpha, 19alpha-trihydroxyurs-12-en-28-oic (7), scopolin (8), scutellarein-7-O-alpha-L-rhamnoside (9), scopoletin (10), quercetin (11), quercetin-3-O-beta-D-glucoside (12), apigenin (13), 2alpha-hydroxyursolic acid (14). All compounds are obtained from this plant for the first time.

  4. Analysis of states in {sup 13}C populated in {sup 9}Be + {sup 4}He resonant scattering

    SciTech Connect

    Freer, M.; Ashwood, N. I.; Curtis, N.; Kokalova, Tz.; Wheldon, C.; Di Pietro, A.; Figuera, P.; Fisichella, M.; Scuderi, V.; Torresi, D.; Grassi, L.; Jelavic Malenica, D.; Koncul, M.; Mijatovic, T.; Prepolec, L.; Skukan, N.; Soic, N.; Szilner, S.; Tokic, V.; Milin, M.

    2011-09-15

    Measurements of {sup 9}Be + {alpha} resonant scattering have been performed using the thick-target approach with a {sup 4}He gas volume and a large-area silicon strip detector. {sup 9}Be beam energies in the range 12 to 21.4 MeV were used to measure the {sup 13}C excitation energy spectrum between 13.2 and 16.2 MeV. An R-matrix analysis has been performed to characterize the spins and widths of {sup 13}C resonances, some of which have been proposed to be associated with a 3{alpha}+n molecular band.

  5. Phenylethanoid and iridoid glycosides from the Thai medicinal plant, Barleria strigosa.

    PubMed

    Kanchanapoom, Tripetch; Noiarsa, Pawadee; Ruchirawat, Somsak; Kasai, Ryoji; Otsuka, Hideaki

    2004-05-01

    A phenylethanoid (4-hydroxyphenylethyl 4-O-beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranoside) and an iridoid (10-O-trans-coumaroyl-eranthemoside) were isolated from an entire Barleria strigosa plant together with verbascoside, isoverbascoside, decaffeoylverbascoside, (+)-lyoniresinol 3alpha-O-beta-D-glucoside, apigenin 7-O-alpha-L-rhamnosyl-(1-->6)-O-beta-D-glucoside, 7-O-acetyl-8-epi-loganic acid and (3R)-1-octen-3-ol-3-O-beta-D-xylosyl-(1-->6)-beta-D-glucoside. The structural elucidations were based on analyses of physical and spectroscopic data.

  6. Lamb shift in the muonic helium ion

    SciTech Connect

    Martynenko, A. P.

    2007-07-15

    The Lamb shift (2P{sub 1/2}-2S{sub 1/2}) in the muonic helium ion ({mu}-{sub 2}{sup 4}He){sup +} is calculated with the account of contributions of orders {alpha}{sup 3}, {alpha}{sup 4}, {alpha}{sup 5}, and {alpha}{sup 6}. Special attention is given to corrections of the electron vacuum polarization, the nuclear structure, and recoil effects. The obtained numerical value of the Lamb shift 1381.72 meV can be considered as a reliable estimate for the comparison with experimental data.

  7. Lamb shift in the muonic deuterium atom

    SciTech Connect

    Krutov, A. A.; Martynenko, A. P.

    2011-11-15

    We present an investigation of the Lamb shift (2P{sub 1/2}-2S{sub 1/2}) in the muonic deuterium ({mu}D) atom using the three-dimensional quasipotential method in quantum electrodynamics. The vacuum polarization, nuclear-structure, and recoil effects are calculated with the account of contributions of orders {alpha}{sup 3}, {alpha}{sup 4}, {alpha}{sup 5}, and {alpha}{sup 6}. The results are compared with earlier performed calculations. The obtained numerical value of the Lamb shift at 202.4139 meV can be considered a reliable estimate for comparison with forthcoming experimental data.

  8. Crosstalk between normal and tumoral brain cells. Effect on sex steroid metabolism.

    PubMed

    Melcangi, R C; Cavarretta, I; Magnaghi, V; Ballabio, M; Martini, L; Motta, M

    1998-02-01

    The present article shows for the first time that two cell lines derived respectively from a rat glioma (C6 cell line) and from a human astrocytoma (1321N1 cell line) are able to convert testosterone and progesterone into their corresponding 5 alpha-reduced metabolites dihydrotestosterone and dihydroprogesterone. Moreover, both cell lines are also able to convert these metabolites further into their corresponding 3 alpha-OH derivatives, 5 alpha-androstan-3 alpha, 7 beta-diol (3 alpha-diol) and tetrahydroprogesterone. On the basis of these observations, the possibility that secretory products of normal and tumoral brain cells might be able to influence steroid metabolism occurring in the two glial cell lines previously mentioned as well as in fetal rat neurons and in neonatal rat type 1 astrocytes has been considered. To this purpose, cultures of the different cellular types have been exposed to the conditioned medium in which the other cells were grown. The results obtained indicate that: 1. Neurons are able to stimulate, in a statistically significant fashion, the formation of dihydrotestosterone (DHT), 3 alpha-diol, and tetrahydraprogesterone (THP) in C6 cells. 2. Type 1 astrocytes, on the contrary, are unable to modify steroid metabolism in C6 cells. 3. C6 cell product(s) decrease(s) the formation of DHP in type 1 astrocytes, without modifying that of DHT. 4. C6 cells do not influence the metabolism of testosterone (T) and progesterone (P) in neurons. In conclusion, the present observations show that the conditioned medium of normal neurons is able to increase the metabolism of testosterone and progesterone occurring in a tumoral glial cell line, and that the conditioned media of the two tumoral cell lines analyzed are able to decrease the conversion of P into DHP occurring in normal type 1 astrocytes. The surprising result that these conditioned media do not alter the formation of DHT is discussed. Work is presently in progress to identify the principle

  9. Experimental autoimmune anti-glomerular basement membrane glomerulonephritis: a protective role for IFN-gamma.

    PubMed

    Kitching, A Richard; Turner, Amanda L; Semple, Timothy; Li, Ming; Edgtton, Kristy L; Wilson, Gabrielle R; Timoshanko, Jennifer R; Hudson, Billy G; Holdsworth, Stephen R

    2004-07-01

    IL-12 and IFN-gamma play key roles in murine lupus and planted antigen models of glomerulonephritis. However, their roles in renal organ-specific autoimmunity are unknown. To establish the roles of endogenous IFN-gamma and IL-12 in experimental autoimmune anti-glomerular basement membrane (GBM) glomerulonephritis (EAG), EAG was induced in normal C57BL/6 mice (WT), IL-12p40-deficient (IL-12p40-/-) mice, and IFN-gamma-deficient (IFN-gamma-/-) mice by immunization with alpha3-alpha5(IV)NC1 heterodimers. At 13 wk, WT mice developed EAG with linear mouse anti-GBM antibody deposition, histologic injury, proteinuria, and mild tubulointerstitial disease. Compared with WT mice, IL-12p40-/- mice had decreased histologic injury and trends to decreased leukocyte infiltrates. In contrast, 40% (4 of 10) of IFN-gamma-/- mice developed significant crescent formation and focal or diffuse interstitial infiltrates (WT, 0 of 8). Compared with WT and/or IL-12p40-/- mice, IFN-gamma-/- mice developed increased injury: histologic injury, total glomerular cell numbers, leukocytes in glomeruli, and renal expression of P-selectin and intercellular adhesion molecule 1. All groups developed similar serum anti-alpha3-alpha5(IV)NC1 antibodies and glomerular Ig deposition, but IFN-gamma-/- mice had decreased anti-alpha3-alpha5(IV)NC1 IgG2a. Therefore, IFN-gamma-/- mice developed increased cellular reactants despite a potentially less damaging antibody response. Dermal delayed-type hypersensitivity was increased in alpha3-alpha5(IV)NC1 immunized IFN-gamma-/- mice and was suppressed by recombinant murine IFN-gamma. CD4+ cells from draining nodes of immunized IFN-gamma-/- mice showed increased proportions of proliferating CD4+ cells but similar numbers of apoptotic cells. These studies demonstrate that in renal organ-specific autoimmunity, IL-12 is pathogenetic but IFN-gamma is protective. They lend weight to the hypothesis that depending on the context/severity of the nephritogenic immune response

  10. Structures of two new fibrinolytic saponins from the seed of Luffa cylindrica Roem.

    PubMed

    Yoshikawa, K; Arihara, S; Wang, J D; Narui, T; Okuyama, T

    1991-05-01

    Two new fibrinolytic saponins, lucyosides N and P, were isolated from the seeds of Luffa cylindrica Roem. (Cucurbitaceae). On the basis of chemical and spectral evidence, lucyoside N was characterized as 3-O-beta-D-galactopyranosyl-(1----2)-beta-D-glucuronopyranosyl-28- O-beta-D-xylopyranosyl-(1----4)-[beta-D-glucopyranosyl-(1----3)]-alpha-L -rhamnopyranosyl-(1----2)-alpha-arabinopyranosyl quillaic acid. Lucyoside P was characterized as a gypsogenin glycoside with the same sugar moiety as lucyoside N.

  11. Structural variation of glycolipids from Meiothermus taiwanensis ATCC BAA-400 under different growth temperatures.

    PubMed

    Yang, Yu-Liang; Yang, Feng-Ling; Huang, Zih-You; Tsai, Yu-Hsuan; Zou, Wei; Wu, Shih-Hsiung

    2010-10-07

    A major glycolipid, alpha-Galf(1-3)-alpha-Galp(1-6)-beta-GlcpNAcyl(1-2)-alpha-Glcp(1-1)-2-acylalkyldiol, is obtained from Meiothermus taiwanensis. This novel glycolipid is found only when the bacterium grows above 62 degrees C, which is significantly different from those from the same bacteria incubated at 55 degrees C. Terminal galactofuranoside and 1,2-alkyldiol lipids replaced galactopyranoside and glycerol lipids, respectively, under increased growth temperature. This variation is likely necessary for bacteria for keeping the stable outer membrane and surviving under extreme environments.

  12. Molecular mechanism of antioxidant synergism of tocotrienols and carotenoids in palm oil.

    PubMed

    Schroeder, Maria T; Becker, Eleonora Miquel; Skibsted, Leif H

    2006-05-03

    During repeated deep-fat frying of potato slices at 163 degrees C in yellow or red palm olein of comparable fatty acid profiles, the oxidative stability (peroxide value and anisidine value) of the palm oleins was similar, and in yellow palm olein, the rate of antioxidant depletion decreased in the order gamma-T3 > alpha-T3 > delta-T3 (T3, tocotrienol). In red palm olein, which had a total tocopherol/tocotrienol content of 1260 vs 940 ppm in yellow palm olein and a corresponding longer induction period in the Rancimat stability test at 120 degrees C, only depletion of gamma-T3 was significant among the phenols during frying and slower as compared to that in yellow palm olein. The carotenes in the red palm olein were depleted linearly with the number of fryings, apparently yielding an overall protection of the phenols. In antioxidant-depleted palm olein and in phospholipid liposomes with added increasing concentrations of phenols, gamma-T3 was found to be a better antioxidant than alpha-T3. alpha-T3 and alpha-T (T, tocopherol) had a similar antioxidant effect in antioxidant-depleted palm olein in the Rancimat stability test, while in the liposomes the ordering as determined by induction period for the formation of conjugated dienes was gamma-T3 > alpha-T3 > alpha-T. The addition of 100-1000 ppm beta-carotene to antioxidant-depleted palm olein or liposomes (lycopene also tested) did not provide any protection against oxidation. In the liposomes, synergistic interactions were observed between beta-carotene or lycopene and alpha-T, alpha-T3, or gamma-T3 for carotene/phenol ratios of 1:10 and 1:2 but not for 1:1. In chloroform, carotenes were regenerated by tocopherols/tocotrienols from carotene radicals generated by laser flash photolysis as shown by transient absorption spectroscopy, suggesting that carotenes rather than phenols are the primary substrate for lipid-derived radicals in red palm olein, in effect depleting carotenes prior to phenols during frying

  13. [Effect of a new derivative of glutamic and apovincaminic acids on brain metabolism in post-ischemic period].

    PubMed

    Makarova, L M; Prikhod'ko, M A; Pogorelyĭ, V E; Skachilova, S Ia; Mirzoian, R S

    2014-01-01

    Neuroprotective properties of the new derivative of glutamic and apovincaminic acids, ethyl -(3-alpha,16-alpha)-eburnamenin-14-carbopxylate of 2-aminopentadionic acid (LHT 1-02) were studied on a model of acute brain ischemia in cats. LHT 1-02 has proved to be more effective than the reference drugs vinpocetin and glycine in preventing the reperfusive damage, which was manifested by decreased postischemic hyperglycemia, activated utilization of oxygen in the brain, and suppressed postischemic metabolic lactate acidosis. Thus, the results of this comparative study show expediency of further investigations of LHT 1 - 02 as a potential neuroprotective drug.

  14. Lanostane triterpenoids from the inedible mushroom Fomitopsis spraguei.

    PubMed

    Quang, Dang Ngoc; Arakawa, Yuuki; Hashimoto, Toshihiro; Asakawa, Yoshinori

    2005-07-01

    Investigation of the methanolic extract of the inedible mushroom Fomitopsis spraguei (Polyporaceae) led to the isolation of five lanostane-type triterpenoids (1-5): three new compounds named fomitopsins A-C (2-4), and two known compounds, quercinic acid C (1) and 3alpha-carboxyacetyl-12beta-hydroxyquercinic acid (5). Their structures were determined by 2D NMR, MS, IR, UV spectra, and X-ray crystallographic analyses. An X-ray crystal structure analysis of quercinic acid C (1) established its stereochemistry as 3R,12R-dihydroxy-24R-methyl-23-oxo-25S-lanost-8-en-26-oic acid.

  15. Plant antitumor agents, 21. Flavones, coumarins, and an alkaloid from Sargentia greggii.

    PubMed

    Meyer, B N; Wall, M E; Wani, M C; Taylor, H L

    1985-01-01

    Investigation of the MeOH extracts of Sargentia greggii (Rutaceae) led to the isolation of two known flavones, zapotin and 5,6,2'-trimethoxyflavone, and a new flavone, 5,6,2',3',4',6'-hexamethoxyflavone (1), whose structure was established by spectral data and confirmed by total synthesis. In addition, two known coumarins, 3-(alpha,alpha-dimethylallyl) herniarin and seselin, and a new coumarin, O-geranylosthenol (3), have been isolated from this plant. The structure of 3 was deduced from spectral data. Although the crude extracts displayed KB activity, none of the crystalline compounds were significantly active.

  16. New evidence that both T-type calcium channels and GABAA channels are responsible for the potent peripheral analgesic effects of 5alpha-reduced neuroactive steroids.

    PubMed

    Pathirathna, Sriyani; Brimelow, Barbara C; Jagodic, Miljen M; Krishnan, Kathiresan; Jiang, Xin; Zorumski, Charles F; Mennerick, Steven; Covey, Douglas F; Todorovic, Slobodan M; Jevtovic-Todorovic, Vesna

    2005-04-01

    Neurosteroids are potent blockers of neuronal low-voltage activated (T-type) Ca(2+) channels and potentiators of GABA(A) ligand-gated channels, but their effects in peripheral pain pathways have not been studied previously. To investigate potential analgesic effects and the ion channels involved, we tested the ability of locally injected 5alpha-reduced neurosteroids to modulate peripheral thermal nociception to radiant heat in adult rats in vivo and to modulate GABA(A) and T-type Ca(2+) channels in vitro. The steroid anesthetic alphaxalone (ALPX), the endogenous neurosteroid allopregnanolone (3alpha5alphaP), and a related compound ((3alpha,5alpha,17beta)-3-hydroxyandrostane-17-carbonitrile, (ACN)), induced potent, dose-dependent, enantioselective anti-nociception in vivo and modulation of both T-type Ca(2+) currents and GABA(A)-mediated currents in vitro. Analgesic effects of ALPX were incompletely antagonized by co-injections of the GABA(A) receptor antagonist bicuculline. The neurosteroid analogue ((3alpha,5alpha)-3-hydroxy-13,24-cyclo-18,21-dinorchol-22-en-24-ol (CDNC24), a compound with GABAergic but not T-type activity, was not analgesic. However, (3beta,5alpha,17beta)-17-hydroxyestrane-3-carbonitrile (ECN)), which has effects on T-type channels but not on GABA(A) receptors, also induced potent enantioselective peripheral anti-nociception. ECN increased pain thresholds less than ALPX, 3alpha5alphaP and ACN. However, when an ineffective dose of CDNC24 was combined with ECN, anti-nociceptive activity was greatly enhanced, and this effect was bicuculline-sensitive. These results strongly suggest that GABA(A) channels do not contribute to baseline pain transmission, but they can enhance anti-nociception mediated by blockade of T-type Ca(2+) channels. In conclusion, we demonstrate that potent peripheral analgesia induced by 5alpha-reduced neurosteroid is mediated in part by effects on T-type Ca(2+) channels. Our results also reveal a role of GABA-gated ion channels

  17. Potential anticancer agents. IV. Constituents of Jacaranda caucana Pittier (Bignoniaceae).

    PubMed

    Ogura, M; Cordell, G A; Farnsworth, R

    1977-01-01

    An aqueous ethanol extract of Jacaranda caucana Pittier (Bignoniaceae) showed in vivo antitumor activity against the P-388 lymphocytic leukemia system. Fractionation, accompanied by monitoring for biological activity, afforded a novel phytoquinoid derivative jacaranone, which exhibited both in vivo antitumor and in vitro cytotoxic activity. beta-Sitosterol, betulinic acid, ursolic acid, 2alpha-hydroxyursolic acid, 2alpha,3alpha-dihydroxyurs-12-en-28-oic acid and a new triterpene acid, jacarandic acid, were also isolated. The structure elucidation of jacarandic acid is described.

  18. Uncertainties in the use of periodate oxidation for determination of dextran structure.

    PubMed

    Leonard, G J

    1975-05-01

    A glucan of high molecular weight isolated from stale sugar-cane, and previously shown to have a marked effect on sucrose crystallisation processes-1, is a relatively linear dextran. Approximately 96-97% of its D-glucose residues are involved in (1 yields 6)-alpha-D linkages and constitute the linear backbone of the polymer. The remaining 3-4% of D-glucose residues form branch-points by (1 yields 3)-alpha-D linkages. The periodate-oxidation technique, which has been extensively used by other workers to determine dextran structure, gave erroneous results when applied to the dextran from stale sugar-cane.

  19. Alkaloid content of the seeds from Erythroxylum Coca var. Coca.

    PubMed

    Casale, John F; Toske, Steven G; Colley, Valerie L

    2005-11-01

    Alkaloid extracts from the seeds of Erythroxylum Coca var. Coca grown in the Chapare Valley of Bolivia were subjected to gas and liquid chromatographic-mass spectrometric analyses. Several alkaloids from these seeds were detected and characterized, including methylecgonidine, tropine, 3alpha-acetoxytropane, ecgonine methyl ester, cuscohygrine, N-norbenzoyltropine, benzoyltropine, hexanoylecgonine methyl ester, cocaine, cis-cinnamoylcocaine, and trans-cinnamoylcocaine. Methylecgonidine was determined to be the primary constituent and not an analytical artifact. Additionally, two significant new uncharacterized alkaloids were established as present. Recent evidence suggests that some cocaine processors are adding this seed extraction material to cocaine extracted from coca leaf and may impact cocaine impurity signature profiles.

  20. Role of distinct type IV collagen networks in glomerular development and function.

    PubMed

    Harvey, S J; Zheng, K; Sado, Y; Naito, I; Ninomiya, Y; Jacobs, R M; Hudson, B G; Thorner, P S

    1998-12-01

    In X-linked Alport syndrome, mutations in the COL4A5 gene encoding the alpha 5 chain of type IV collagen result in progressive renal failure. This nephropathy appears to relate to the arrest of a switch from an alpha 1/alpha 2 to an alpha 3/alpha 4/alpha 5 network of type IV collagen in the developing glomerular basement membrane (GBM; Kalluri et al, J Clin Invest 99:2470, 1997). We examined the role of this switch in glomerular development and function using a canine model of X-linked nephritis with a COL4A5 mutation. The electron microscopic appearance and the expression of the alpha 1-alpha 6 chains of type IV collagen in the GBM was correlated with glomerular function. In normal neonatal glomeruli, once capillary loops were present, there was staining of GBM for the alpha 1-alpha 5 chains. Prior to this stage, only alpha 1 and alpha 2 chains were present, with rare glomeruli positive for the alpha 5 chain. As glomeruli matured, the alpha 1 and alpha 2 chains tended to disappear from the GBM, with the alpha 3-alpha 5 chains remaining. In affected male dogs, only the alpha 1 and alpha 2 chains were detected at any stage. GBM ultrastructure in these dogs remained normal until one month and proteinuria did not appear until two months. Our results show that normal glomerular development involves a switch in type IV collagen networks. In affected male dogs, a failure of this switch results in an absence of the alpha 3/alpha 4/alpha 5 network and a persistence of the alpha 1/alpha 2 network in GBM. GBM ultrastructure and glomerular function remain normal for one month, indicating that GBM deterioration in Alport syndrome begins as a postnatal process. Hence, only the alpha 1/alpha 2 network is essential for normal glomerular development, whereas the alpha 3/alpha 4/alpha 5 network is essential for long-term maintenance of glomerular structure and function.

  1. Silicon solar cell testing in concentrated sunlight and simulated sunlight

    NASA Technical Reports Server (NTRS)

    Burgess, E. L.; Mitchell, K. W.

    1976-01-01

    A method is described for testing silicon solar cells in concentrated sunlight and simulated sunlight. Concentrated sunlight is obtained by using an acrylic Fresnel lens; the simulated sunlight source is a short arc Xenon lamp. Average illumination levels during the tests are inferred from an assumed linear relationship between short circuit current and illumination. The linearity assumption is investigated for 0.3 alpha cm base resistivity silicon cells and found to be valid. Some typical results are presented to illustrate the type of information obtained during the testing.

  2. A Review for the US Navy of Best Practices, Knowledge and Data Gaps, and Research Directions for Vapor Intrusion

    DTIC Science & Technology

    2009-05-01

    0 0 Ju n- 01 D ec - 0 1 Ju n- 02 D ec - 0 2 Ju n- 03 D ec - 0 3 Ju n- 04 D ec - 0 4 Ju n...05 D ec - 0 5 Ju n- 06 D ec - 0 6 Ju n- 07 D ec - 0 7 Sampling Date C o n c e n tr a ti o n ( p p b v ) SVOC 1 SVOC 2 SVOC 3 SVOC 4 PCE TCE All TCE...A ha= 1.0 ---Alpha= 1&1 --- - Atpha = 1& 2 --- · - Alpha= 1E-3 ----Alpha=

  3. Attenuation of mild hyperandrogenic activity in postpubertal acne by a triphasic oral contraceptive containing low doses of ethynyl estradiol and d,l-norgestrel.

    PubMed

    Lemay, A; Dewailly, S D; Grenier, R; Huard, J

    1990-07-01

    The effect of a low dose triphasic oral contraceptive (OC) was evaluated during a 6-month treatment period in 41 patients (mean age, 25.4 +/- 0.7 yr) who had grade I-IV postpubertal acne and normal menses. The OC contained three dose levels of ethynyl estradiol and dl-norgestrel. Acne lesions were assessed, and serum androgen levels were measured during a control cycle and between days 17-21 of treatment cycles 1, 2, 3, and 6. Four patients dropped out after 3 months of treatment. Acne was significantly improved after the first OC cycle. After six cycles, the number of comedones had decreased by 79.6 +/- 3.2% (range, 50-100%) in 69.4% of the patients. Mean baseline levels of testosterone, 17-hydroxyprogesterone, and dehydroepiandrosterone sulfate were in the upper third of the normal range, with elevated individual values in 18.9%, 36.5%, and 26.8% of the women, respectively. Mean baseline levels of androstenedione, free testosterone (T), and 3 alpha-androstanediol glucuronide (3 alpha-diol-G) were above the normal range, with elevated individual values in 51.2%, 75.0%, and 85.4% of the patients, respectively. Sex hormone-binding globulin (SHBG) levels were below the normal range in 26.8% of the cases. At the end of the first OC cycle, there was a significant (P less than 0.01) decrease in all androgen precursors and a 2-fold increase in SHBG. Androstenedione and free T decreased into the normal range during OC intake. Serum 3 alpha-diol-G levels remained elevated, but had decreased by 34.5% at cycle 6 (P less than 0.05). These results show that the triphasic OC has significantly improved acne in postpubertal women for whom acne was the main manifestation of mild hyperandrogenic activity. The improvement in acne corresponded to a decrease in adrenal/ovarian androgens and free T, which led to a decreased metabolism to 3 alpha-diol-G, presumably by the sebaceous glands. The increase in SHBG is considered an estrogenic effect, and the triphasic formulation containing

  4. Differential expression of beta 1 integrins in nonneoplastic smooth and striated muscle cells and in tumors derived from these cells.

    PubMed Central

    Mechtersheimer, G.; Barth, T.; Quentmeier, A.; Möller, P.

    1994-01-01

    Integrins are a superfamily of transmembrane alpha beta heterodimers that play an important role in cell-matrix and cell-cell interactions by acting as receptors for extracellular matrix proteins and for cell adhesion molecules. Using monoclonal antibodies against beta 1, alpha 1 to alpha 6, and alpha v subunits, the in situ distribution pattern of beta 1 integrins was examined immunohistochemically in nonneoplastic smooth and striated muscle cells and in their tumors. Nonneoplastic smooth muscle cells were beta 1+, alpha 1+, alpha 3+, alpha v+ and, in diverse localizations, also alpha 5+ or even alpha 6+. The expression of the beta 1 chain was conserved in all leiomyomas and leiomyosarcomas. The distribution pattern of the alpha subunits by contrast underwent several changes during malignant transformation of smooth muscle cells. These alterations consisted in a neoexpression of alpha 2, alpha 4, and alpha 6 as well as in an abnormal abrogation of alpha 1 and alpha 3 in some leiomyosarcomas. Except for the absence of alpha 5 in the majority of epithelioid leiomyosarcomas, expression of the alpha 5 and alpha v subunits was mainly conserved. In addition, tumors with epithelioid differentiation differed from typical cases by the absence of alpha 1 and the simultaneous presence of alpha 4. Adult striated muscle cells were beta 1+ but alpha 1- to alpha 6- and alpha v-, whereas fetal striated muscle cells were not only beta 1+ but also alpha 3+/-, alpha 4+/-, alpha 5+ and alpha 6+. In all rhabdomyosarcomas the expression of beta 1 was retained. Furthermore, the majority of cases showed the expression of one or more alpha subunits most of which, ie, alpha 4, alpha 5, and alpha 6, were also found in fetal striated muscle cells. In conclusion, beta 1 integrins exhibited a differential expression pattern along the two lines of myogenic differentiation. This integrin profile underwent characteristic changes during malignant transformation. Nevertheless, the compiled

  5. Effect of testosterone metabolites on ABC half-transporter relative gene expression in X-linked adrenoleukodystrophy.

    PubMed

    Petroni, A; Cappa, M; Carissimi, R; Blasevich, M; Uziel, G

    2007-10-01

    X-linked adrenoleukodystrophy (X-ALD) is an inherited neurodegenerative disorder associated with reduced very long-chain fatty acid beta-oxidation, mainly affecting the nervous system, the adrenal cortex and the testes. The clinical manifestations of hypogonadism, alopecia and the impairment of the enzyme 5alpha-reductase, which converts testosterone into dihydrotestosterone, clearly point to an involvement of androgens in this pathology. The disease is characterized by mutations in the ABCD1 gene, which codes for the peroxisomal ABC half-transporter ALDP, and by a broad range of clinical manifestations. The altered function of ALDP can be compensated by the overexpression of proteins belonging to the same family of ABC half-transporters. A promising therapeutic approach is represented by the activation of these proteins by specific agonists. In this study we evaluated the effect of the testosterone metabolite dihydrotestosterone (DHT) and 5alpha-androstan-3alpha,17beta-diol (3alpha-diol) on the expression of the ABC half-transporters encoded by the ABCD2 and ABCD3 genes, in fibroblasts drawn from controls and from two affected brothers. The two patients presented the same mutation in exon 9 but had different clinical manifestations, one patient being asymptomatic and the second one severely affected. When the cells were stimulated with testosterone metabolites, only the severely affected patient showed a significant increase in ABCD2 mRNA levels, while the ABCD3 expression remained unchanged in both patients.

  6. Urinary metabolomics in Fxr-null mice reveals activated adaptive metabolic pathways upon bile acid challenge.

    PubMed

    Cho, Joo-Youn; Matsubara, Tsutomu; Kang, Dong Wook; Ahn, Sung-Hoon; Krausz, Kristopher W; Idle, Jeffrey R; Luecke, Hans; Gonzalez, Frank J

    2010-05-01

    Farnesoid X receptor (FXR) is a nuclear receptor that regulates genes involved in synthesis, metabolism, and transport of bile acids and thus plays a major role in maintaining bile acid homeostasis. In this study, metabolomic responses were investigated in urine of wild-type and Fxr-null mice fed cholic acid, an FXR ligand, using ultra-performance liquid chromatography (UPLC) coupled with electrospray time-of-flight mass spectrometry (TOFMS). Multivariate data analysis between wild-type and Fxr-null mice on a cholic acid diet revealed that the most increased ions were metabolites of p-cresol (4-methylphenol), corticosterone, and cholic acid in Fxr-null mice. The structural identities of the above metabolites were confirmed by chemical synthesis and by comparing retention time (RT) and/or tandem mass fragmentation patterns of the urinary metabolites with the authentic standards. Tauro-3alpha,6,7alpha,12alpha-tetrol (3alpha,6,7alpha,12alpha-tetrahydroxy-5beta-cholestan-26-oyltaurine), one of the most increased metabolites in Fxr-null mice on a CA diet, is a marker for efficient hydroxylation of toxic bile acids possibly through induction of Cyp3a11. A cholestatic model induced by lithocholic acid revealed that enhanced expression of Cyp3a11 is the major defense mechanism to detoxify cholestatic bile acids in Fxr-null mice. These results will be useful for identification of biomarkers for cholestasis and for determination of adaptive molecular mechanisms in cholestasis.

  7. The structure of the O-specific polysaccharide chain of the lipopolysaccharide of Salmonella arizonae O61.

    PubMed

    Vinogradov, E V; Shashkov, A S; Knirel, Y A; Kochetkov, N K; Dabrowski, J; Grosskurth, H; Stanislavsky, E S; Kholodkova, E V

    1992-07-02

    The O-specific polysaccharide was obtained by mild degradation of the Salmonella arizonae O61 lipopolysaccharide with acid. It contained 2-acetamido-2-deoxy-D-glucose, 2-acetamidino-2,6-dideoxy-L-galactose (FucAm), and 7-acetamido-3,5,7,9-tetradeoxy-5-[(R)-3-hydroxybutyramido]-D- glycero-L-galacto-nonulosonic acid (Sug). On the basis of partial acid hydrolysis with 0.1 M HCl, solvolysis with anhydrous HF in methanol, and 1H- and 13C-NMR analysis (including 1H/13C inversely correlated spectroscopy for localisation of N-acyl substituents), it was concluded that the O-specific polysaccharide had the following structure. ----3)-alpha-L-FucAm-(1----3)-alpha-D-GlcNAc-(1----8)-beta-Sug+ ++-(2---- The O-antigen of S. arizonae O61 is structurally related to that of Pseudomonas aeruginosa O12, thus explaining the known serological cross-reactivity between these micro-organisms.

  8. Sexually dimorphic effects of postnatal allopregnanolone on the development of anxiety behavior after early deprivation.

    PubMed

    Zimmerberg, Betty; Kajunski, Elizabeth W

    2004-07-01

    Stress early in life exerts persistent detrimental effects on brain development. In this experiment, a rodent model of child neglect, early deprivation (ED), was used to investigate the role of the neurosteroid allopregnanolone [AlloP; 3alpha-hydroxy-5alpha-pregnan-20-one (3alpha,5alpha-THP)] in the development of anxiety behavior. Subjects were either undisturbed controls or ED: separated individually for 6 h per day from postnatal day (PN) 2 to 6. Control and ED subjects were also either noninjected, vehicle-injected or injected with 5 mg/kg AlloP prior to the isolation. At PN 7, responses to 2.5 or 5 microg icv injections of AlloP were determined for separation-induced ultrasonic vocalizations (USVs). Tolerance to the USV-reducing effect of daily AlloP was seen in control but not ED pups, and daily AlloP reversed the expected ED suppression of USVs. As adults, controls treated with postnatal AlloP were less anxious than all other groups on the elevated plus maze. ED counteracted this effect. Male controls showed a reversal of the typical sex difference. There were no effects on open-field activity. These results suggest that the neonatal brain is responsive to alterations in AlloP levels, and that neuroactive progesterone metabolites may play a role in mediating the development of stress-related sex differences.

  9. Structure of the O-polysaccharide and serological studies of the lipopolysaccharide of Proteus penneri 60 classified into a new Proteus serogroup O70.

    PubMed

    Zych, Krystyna; Perepelov, Andrei; Baranowska, Agata; Zabłotni, Agnieszka; Shashkov, Alexander S; Knirel, Yuriy A; Sidorczyk, Zygmunt

    2005-03-01

    An alkali-treated lipopolysaccharide of Proteus penneri strain 60 was studied by chemical analyses and 1H, 13C and 31P NMR spectroscopy, and the following structure of the linear pentasaccharide-phosphate repeating unit of the O-polysaccharide was established: 6)-alpha-D-Galp-(1-->3)-alpha-L-FucpNAc-(1-->3)-alpha-D-GlcpNAc-(1-->3)-beta-D-Quip4NAc-(1-->6)-alpha-D-Glcp-1-P-(O--> Rabbit polyclonal O-antiserum against P. penneri 60 reacted with both core and O-polysaccharide moieties of the homologous LPS. Based on the unique O-polysaccharide structure and serological data, we propose to classify P. penneri 60 into a new, separate Proteus serogroup O70. A weak cross-reactivity of P. penneri 60 O-antiserum with the lipopolysaccharide of Proteus vulgaris O8, O15 and O19 was observed and discussed in view of the chemical structures of the O-polysaccharides.

  10. Structure of the O-specific polysaccharide of Proteus vulgaris O15 containing a novel regioisomer of N-acetylmuramic acid, 2-acetamido-4-O-[(R)-1-carboxyethyl]-2-deoxy-D-glucose.

    PubMed

    Perepelov, Andrei V; Torzewska, Agnieszka; Shashkov, Alexander S; Ziolkowski, Andrzej; Senchenkova, Sof'ya N; Rozalski, Antoni; Knirel, Yuriy A

    2002-11-29

    An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of Proteus vulgaris O15 and studied by sugar and methylation analyses along with 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, ROESY, and H-detected 1H,(13)C HMQC experiments. The polysaccharide was found to contain an ether of GlcNAc with lactic acid, and the following structure of the repeating unit was established:-->3)-alpha-D-GlcpNAc4(R-Lac)6Ac-(1-->2)-beta-D-GlcpA-(1-->3)-alpha-L-6dTalp2Ac-(1-->3)-beta-D-GlcpNAc-(1-->where L-6dTal and D-GlcNAc4(R-Lac) are 6-deoxy-L-talose and 2-acetamido-4-O-[(R)-1-carboxyethyl]-2-deoxy-D-glucose, respectively. The latter sugar, which to our knowledge has not been hitherto found in nature, was isolated from the polysaccharide by solvolysis with anhydrous triflic acid and identified by comparison with the authentic synthetic compound. Serological studies with the Smith-degraded polysaccharide showed an importance of 2-substituted GlcA for manifesting of the immunospecificity of P. vulgaris O15.

  11. Location of acyl groups of trehalose-containing lipooligosaccharides of mycobacteria.

    PubMed Central

    Camphausen, R T; McNeil, M; Jardine, I; Brennan, P J

    1987-01-01

    A variant of a Mycobacterium sp. originating in a patient with Crohn's disease, but not necessarily implicated in the disease, provided a simple version of a newer class of species-specific surface glycolipids, the trehalose-containing lipooligosaccharides. A combination of high-resolution 1H nuclear magnetic resonance, methylation, ethylation, and absolute configurational analysis established the structure of the oligosaccharide unit as beta-D-Glcp(1----3)-alpha-L-Rhap(1----3)-alpha-D-Glcp(1----1)-alph a-D-Glcp (where Glc is glucose, Rha is rhamnose, and p is pyranosyl), and gas chromatography-electron impact mass spectrometry allowed identification of the fatty acyl esters as primarily 2,4-dimethyltetradecanoate. The relative simplicity of the glycolipid combined with the application of a mild methylation procedure and californium-252 plasma desorption mass spectrometry allowed recognition of three such acyl residues on the 3-, 4-, and 6-hydroxyl positions of the terminal glucosyl residue of the trehalose unit. Thus, the glycolipid is decidedly amphipathic yet is clearly not membranous. This observation leads to speculation about the role of these novel lipooligosaccharides in contributing to the outer segment of the hydrophobic barrier of the cell wall of certain mycobacteria. Images PMID:3680168

  12. The cynomolgus monkey (Macaca fascicularis) is the best animal model for the study of steroid glucuronidation.

    PubMed

    Barbier, Olivier; Bélanger, Alain

    2003-06-01

    Intense research efforts performed during the past decade clearly established the major role of glucuronidation and uridine-diphospho-glucuronosyltransferase (UGT) enzymes for steroid metabolism in humans. However, a clear understanding of the physiological importance of this metabolic process requires in vivo studies. Numerous evidences ascertain that simians are the most appropriate animal models for such studies. Indeed human and monkey have a similar pattern of steroidogenesis, unlike common laboratory mammals such as rat or mouse. Furthermore, human and monkey are unique in having high levels of circulating androsterone glucuronide and androstane-3alpha-diol glucuronide (3alpha-Diol-G). In addition, characterization of eight monkey UGT proteins demonstrated the similarity of their conjugation activity toward steroid hormones. Like human ones, monkey enzymes are expressed in steroid target tissues, where they preferentially glucuronidate androgen and estrogen metabolites. In monkey tissues, immunohistochemical studies demonstrated that UGT2B proteins are expressed in a cell-type specific manner in ovary and kidney, where they control androgens and aldosterone inactivation. These results identify the cynomolgus monkey as an appropriate animal model for the determination of cellular localization of UGT enzymes in steroid target tissues and for the identification of endogenous or exogenous stimuli affecting steroid glucuronidation.

  13. [Metabolism of sex steroid hormones in neuroendocrine organs. I. In vitro androgen metabolism in the hypophyses and epiphyses of rats during the puberal period].

    PubMed

    Degtiar', V G; Pavlinov, S A; Loseva, L A; Isachenkov, V A

    1977-01-01

    Metabolism of tritium-labeled testosterone, delta4-androstane-3, 17-dion, dihydrotestosterone. 5alpha-androstane-3alpha, 17beta-diod and 5alpha-androstane-3beta, 17beta-dion by the isolated hypophyses and epiphyses of male and female 30-day rats was studied. The glands were incubated in Eagle's medium containing the mentioned androgens with the concentration of 1-2.10(-8) M for 90 min at 37 degrees C. As revealed, restoration of testosterone and delta4 androstane-3, 17-dion by 5alpha-reductase occurred in the epiphyses and hypophyses; under conditions of incubation androgen hydroxylation was realized only in the 3alpha- and 17beta-positions. Organ and sexual specificity of the androgen metabolism in the hypophyses and the epiphyses was quantitative in character: the metabolism level of all the androgens was greater in female rats than in the male both in the hypophysis and in the epiphyses; the activity of 5-reductase was lower in the epiphyses of animals of the both sexes than in the hypophyses. It appeared that in the process of biotransformation androgens in the organs under study served as precursors of the polar unidentified metabolites poorly retained by the tissues and eliminated from the organs into the incubation medium during the incubation.

  14. Antifungal activity of the extracts and saponins from Sapindus saponaria L.

    PubMed

    Tsuzuki, Joyce K; Svidzinski, Terezinha I E; Shinobu, Cristiane S; Silva, Luiz F A; Rodrigues-Filho, Edson; Cortez, Diógenes A G; Ferreira, Izabel C P

    2007-12-01

    Extracts from the dried pericarp of Sapindus saponaria L. (Sapindaceae) fruits were investigated for their antifungal activity against clinical isolates of yeasts Candida albicans and C. non-albicans from vaginal secretions of women with Vulvovaginal Candidiasis. Four clinical isolates of C. albicans, a single clinical isolated of each of the species C. parapsilosis, C. glabrata, C. tropicalis, and the strain of C. albicans ATCC 90028 were used. The hydroalcoholic extract was bioactivity-directed against a clinical isolate of C. parapsilosis, and showed strong activity. The n-BuOH extract and one fraction showed strong activity against all isolates tested. Further column-chromatography on silica gel separation of this fraction afforded two pure triterpene acetylated saponins: 3-O-(4-acetyl-beta-D-xylopyranosyl)-(1->3)-alpha-L-rhamnopyranosyl-(1->2)-alpha-L-arabinopyranosyl-hederagenin (1) and 3-O-(3,4-di-acetyl-beta-D-xylopyranosyl)-(1->3)-alpha-L-rhamnopyranosyl-(1->2)-alpha-L-arabynopyranosyl-hederagenin (2). The structures of the compounds were based on spectral data ((1)H and 13C NMR, HSQC, HMBC and MS), and on with literature. The saponins isolated showed strong activity against C. parapsilosis.

  15. Expression of PGE2 EP3 receptor subtypes in the mouse preoptic region.

    PubMed

    Vasilache, Ana Maria; Andersson, Josefin; Nilsberth, Camilla

    2007-08-23

    Inflammatory-induced fever is dependent on prostaglandin E(2) (PGE(2)) binding to its EP(3) receptor in the thermoregulatory region of the hypothalamus, but it is not known which EP(3) receptor isoform(s) that is/are involved. We identified the EP(3) receptor expression in the mouse preoptic region by in situ hybridization and isolated the corresponding area by laser capture microdissection. Real-time RT-PCR analysis of microdissected tissue revealed a predominant expression of the EP(3alpha) isoform, but there was also considerable expression of EP(3gamma), corresponding to approximately 15% of total EP(3) receptor expression, whereas EP(3beta) was sparsely expressed. This distribution was not changed by immune challenge induced by peripheral administration of LPS, indicating that EP(3) receptor splicing and distribution is not activity dependent. Considering that EP(3alpha) and EP(3gamma) are associated with inhibitory and stimulatory G-proteins, respectively, the present data demonstrate that the PGE(2) response of the target neurons is intricately regulated.

  16. [Triterpenoid saponins from flower bud of Jasminum officinale var. grandiflorum].

    PubMed

    Zhao, Gui-Qin; Dong, Jun-Xing

    2008-01-01

    To study the chemical constituent bud of the flowers of Jasminum officinale var. grandiflorum. The compounds were isolated and purified by recrystallization and chromatography on silica gel and Sephadex LH - 20 column. Their structures were elucidated on the basis of physicochemical properties and spectral analysis. Six triterpenoid saponins were identified as 3-O-alpha-L-rhamnopyranosyl (1 --> 2)-beta-D-xylopyranosyl- hederagenin-28-O-beta-D-galactopyranosyl (1 --> 6)-beta-D-galactopyranosyl ester (1), hederagenin-3-O-beta-D-glucopyranosyl (1 --> 3)-alpha-L-arabinopyranoside (2), 2alpha, 3beta, 23-trihydroxyolean-12-en-28-oic-O-beta-D-glucopyranosyl ester (3), hederagenin-3-O-beta-D-xylopyranosyl (1 --> 3)-alpha-L-rhamnopyranosyl (1 --> 2)-alpha-L-arabinopyranoside (4), 2alpha, 3beta, 23-trihydroxyolean-12-en-28-oic-O-alpha-L-rhamnopyranosyl (1 --> 4)-beta-D-glucopyranosyl (1 --> 6)-beta-D-glucopyranosyl ester (5), hederagenin-3-O-alpha-L-rhamnopyranosyl (1 --> 2)-alpha-L-arabinopyranoside (6). Compound 1 is a new compound. Compounds 2, 3, 4, 5, 6 were isolated from the genus Jasminum for the first time.

  17. Improved expression of His(6)-tagged strictosidine synthase cDNA for chemo-enzymatic alkaloid diversification.

    PubMed

    Yang, Liuqing; Zou, Hongbin; Zhu, Huajian; Ruppert, Martin; Gong, Jingxu; Stöckigt, Joachim

    2010-04-01

    Strictosidine synthase (STR1) catalyzes the stereoselective formation of 3alpha(S)-strictosidine from tryptamine and secologanin. Strictosidine is the key intermediate in the biosynthesis of 2,000 plant monoterpenoid indole alkaloids, and it is a key precursor of enzyme-mediated synthesis of alkaloids. An improved expression system is described which leads to optimized His(6)-STR1 synthesis in Escherichia coli. Optimal production of STR1 was achieved by determining the impact of co-expression of chaperones pG-Tf2 and pG-LJE8. The amount and activity of STR1 was doubled in the presence of chaperone pG-Tf2 alone. His(6)-STR1 immobilized on Ni-NTA can be used for enzymatic synthesis of strictosidines on a preparative scale. With the newly co-expressed His(6)-STR1, novel 3alpha(S)-12-azastrictosidine was obtained by enzymatic catalysis of 7-azatryptamine and secologanin. The results obtained are of significant importance for application to chemo-enzymatic approaches leading to diversification of alkaloids with novel improved structures.

  18. Partial characterization of the steroidsulfatases in Peptococcus niger H4.

    PubMed Central

    Van Eldere, J; Parmentier, G; Asselberghs, S; Eyssen, H

    1991-01-01

    The strictly anaerobic intestinal Peptococcus niger H4 synthesizes three different steroidsulfatase enzymes: a constitutive arylsulfatase and two inducible alkylsteroidsulfatases. The arylsulfatase desulfates estrogen-3-sulfates and phenylsulfates. The two alkylsteroidsulfatases desulfate, respectively, 3 alpha-sulfates and 3 beta-sulfates of delta 5, 5 alpha, and 5 beta androstanes, pregnanes, and bile acids. Cholesterol-3 beta-sulfate was not desulfated by the alkylsteroidsulfatases nor were steroids or bile acids that were sulfated in positions other than the 3 position. The alkylsteroidsulfatases were induced by their substrates; bile acid sulfates, however, were poor inducers of the 3 beta-sulfatase and did not induce the 3 alpha-sulfatase activity. In intact bacterial cells, taurine and sulfite suppressed the induction of the alkylsteroidsulfatases and inhibited the activity of the arylsulfatase and alkylsteroidsulfatases. In cell homogenates, the arylsulfatase and alkylsteroidsulfatases activities were inhibited by sulfite and sulfate but not by taurine. Our results support the hypothesis that the main function of the steroidsulfatases in P. niger H4 is to provide the bacteria with sulfur for dissimilatory purposes. PMID:2036022

  19. Efficacy of various natural and synthetic androgens to induce ductal branching morphogenesis in the developing anterior rat prostate.

    PubMed

    Foster, B A; Cunha, G R

    1999-01-01

    The studies presented herein quantitated ductal branching morphogenesis in the anterior prostate (AP) of the newborn rat. Four parameters were measured: epithelial area, epithelial perimeter, node number, and form factor. Nine natural and synthetic androgens were tested for their effectiveness in inducing postnatal prostatic development using 808 newborn rat APs in 68 dose-response experiments. Based on these studies it was shown that testosterone (T) was slightly more effective than dihydrotestosterone (DHT) in supporting ductal branching morphogenesis in the developing rat AP. Furthermore, the activity of T could not be accounted for simply by conversion of T to DHT. Synthetic androgens, 7alpha-methyl-19-nortestosterone and methyltrienolone (R1881), which cannot be 5alpha-reduced to DHT, also induced extensive ductal branching and elicited responses less than those to T and not statistically different from those to DHT. This suggests that although DHT is sufficient for prostatic development, it is not necessary for postnatal ductal branching morphogenesis and growth of the prostate. 5Alpha-androstan-3alpha,17beta-diol was particularly potent in inducing ductal branching, eliciting a response greater than or comparable to those of T and DHT. Androsterone, androstanedione, 5alpha-androstan-3beta,17beta-diol and 5beta-androstan-3alpha,17beta-diol induced ductal branching, but to a lesser extent than either T or DHT. These studies challenge the assumption that DHT is essential for prostatic development, specifically during ductal branching morphogenesis of the neonatal rat prostate.

  20. Testosterone metabolism of fibroblasts grown from prostatic carcinoma, benign prostatic hyperplasia and skin fibroblasts

    SciTech Connect

    Schweikert, H.U.; Hein, H.J.; Romijn, J.C.; Schroeder, F.H.

    1982-02-01

    The metabolism of (1,2,6,7-3H)testosterone was assessed in fibroblast monolayers derived from tissue of 5 prostates with benign hyperplasia (BPH), 4 prostates with carcinoma (PC), and 3 biopsy samples of skin, 2 nongenital skin (NG) and 1 genital skin. The following metabolites could be identified: androstanedione androstenedione, dihydrotestosterone, androsterone, epiandrosterone, androstane-3 alpha, 17 beta-diol and androstane-3 beta, 17 beta-diol. Testosterone was metabolized much more rapidly in fibroblasts originating from prostatic tissue than in fibroblasts derived from NG. A significantly higher formation of 5 alpha-androstanes and 3 alpha-hydroxysteroids could be observed in fibroblasts from BPH as compared to PC. 17-ketosteroid formation exceeded 5 alpha-androstane formation in BPH, whereas 5 alpha-reduction was the predominant pathway in fibroblasts grown from PC and NG. Since testosterone metabolism in fibroblasts of prostatic origin therefore resembles in many aspects that in whole prostatic tissue, fibroblasts grown from prostatic tissues might be a valuable tool for further investigation of the pathogenesis of human BPH and PC.

  1. Effect of supplemental dietary zinc on the mammalian target of rapamycin (mTOR) signaling pathway in skeletal muscle and liver from post-absorptive mice.

    PubMed

    McClung, James P; Tarr, Tyson N; Barnes, Brian R; Scrimgeour, Angus G; Young, Andrew J

    2007-07-01

    Zinc (Zn) is an essential trace element that functions in cellular signaling. The mammalian target of rapamycin (mTOR) regulates the initiation of protein synthesis. The objective of this study was to determine whether Zn could stimulate protein phosphorylation in the mTOR pathway in vivo. Mice (C57BL/6J, n = 30) were fed Zn marginal diets (ZM, 5 mg/kg) for 4 weeks, followed by fasting (F) and/or refeeding with ZM or Zn supplemental (300 mg/kg, ZS) diets for 3 or 6 h. Plasma insulin was greater (P < 0.05) in refed animals as compared to F animals. Protein phosphorylation was detected using multiplex analysis and Western blotting. Multiplex analysis indicated greater (P < 0.05) p70 S6 kinase (p70S6K) and glycogen synthase kinase 3 (GSK-3 alpha/beta) phosphorylation in livers from 6-h refed ZS animals as compared to F animals. Western blots indicated increased (P < 0.05) Akt (Ser 473) phosphorylation in skeletal muscle from animals refed ZS diets for 3 and 6 h as compared to F animals. The ZS diet affected phosphorylation of GSK-3 (alpha/beta) in liver, as 3-h ZS refed animals had greater (P < 0.01) phosphorylation than F animals. These findings indicate that Zn may contribute to the initiation of protein synthesis as a signaling molecule in vivo.

  2. Anti-influenza sesquiterpene from the roots of Reynoutria japonica.

    PubMed

    Nhiem, Nguyen Xuan; Van Kiem, Phan; Van Minh, Chau; Hoai, Nguyen Thi; Duc, Ho Viet; Tai, Bui Huu; Quang, Tran Hong; Le Anh, Hoang Tuan; Yeo, Sang-Gu; Song, Jae-Hyoung; Cheon, Doo-Sung; Park, Moon Ho; Ko, Hyun-Jeong; Kim, Seung Hyun

    2014-03-01

    One new flavonol glycoside, 4'-O-methylmyricitrin 3'-O-beta-D-glucopyranoside (1), one new sesquiterpene, reynoudiol (11), as well as the 12 known compounds (2-10, 12-14) quercetin 3-O-methyl ether (2), quercitrin (3), isorhamnetin 3-alpha-L-rhamnopyranoside (4), tamarixetin 3-alpha-L-rhamnopyranoside (5), myricitrin (6), 4'-O-methylmyricitrin (7), isorhamnetin 3-O-beta-D-xylopyranosyl (1-2)-O-beta- D-glucopyranoside (8), isorhamnetin 3-O-beta- D-apiofuranosyl(1-2)-O-beta- D-glucopyranoside (9), (+)-catechin (10), 7-drimene-3,11,12-triol (12), clovane-2 beta,9 alpha-diol (13), and a-cadinol (14), were isolated from the methanol extract of Reynoutria japonica roots. Based on in vitro screening of the anti-influenza activity of the isolated compounds, reynoudiol showed significantly higher activity than that of oseltamivir phosphate at the same concentration, and did not induce any detectable cytopathic effect in MDCK cells. The CC50 of reynoudiol was above 50 micro M and could inhibit influenza virus infection with an IC50 of 0.29 +/- 0.01 microM. The therapeutic index (TI) of reynoudiol against influenza infection was 172.4, and thus, this compound can be potentially used to treat oseltamivir-resistant influenza virus infection.

  3. Biosynthesis of androgens and pheromonal steroids in neonatal porcine testicular preparations.

    PubMed

    Kwan, T K; Orengo, C; Gower, D B

    1985-04-22

    The biosynthesis of testosterone and 4-androstene-3,17-dione and some 16-androstenes has been studied in homogenates or subcellular fractions of testes from 3-week-old Landrace piglets. Pregnenolone was converted into 5,16-androstadien-3 beta-ol, 4,16-androstadien-3-one, 5 alpha-androst-16-en-3-one and 5 alpha-androst-16-en-3 alpha- and 3 beta-ols, but the quantities were some 50 times less than those formed in the mature boar testis. Androgens were also formed in the microsomal fractions but the quantities of 4-androstene-3,17-dione (from side-chain cleavage of 17-hydroxyprogesterone) and of testosterone (from reduction of 4-androstene-3,17-dione) were 50-70 times lower than in the adult animal. The kinetic parameters and cofactor preference of the 3 alpha- and 3 beta-hydroxysteroid dehydrogenases were determined in the cytosolic, microsomal and mitochondrial fractions of neonatal porcine testes.

  4. Pentacyclic triterpenoids from the aerial parts of Lantana camara and their nematicidal activity.

    PubMed

    Begum, Sabira; Zehra, Syeda Qamar; Siddiqui, Bina Shaheen; Fayyaz, Shahina; Ramzan, Musarrat

    2008-09-01

    Two new olean-12-ene triterpenoids, camarolic acid (1) and lantrigloylic acid (2), have been isolated from the aerial parts of Lantana camara, along with ten known triterpenes, namely, camaric acid, lantanolic acid, lantanilic acid, pomolic acid, camarinic acid, lantoic acid, camarin, lantacin, camarinin, and ursolic acid. The new compounds have been characterized as 3,25-epoxy-3alpha-hydroxy-22beta-{[(S)-3-hydroxy-2-methylidenebutanoyl]oxy}olean-12-en-28-oic acid (1) and 3,25-epoxy-3alpha-hydroxy-22beta-[(3-methylbut-2-enoyl)oxy]olea-9(11),12-dien-28-oic acid (2) through spectroscopic studies and a chemical transformation. Seven of the constituents, namely pomolic acid, lantanolic acid, lantoic acid, camarin, lantacin, camarinin, and ursolic acid, were tested for nematicidal activity against root-knot nematode Meloidogyne incognita. Pomolic acid, lantanolic acid, and lantoic acid showed 100% mortality at 1 mg/ml concentration after 24 h, while camarin, lantacin, camarinin, and ursolic acid exhibited 100% mortality at this concentration after 48 h. These results are comparable to those obtained with the conventional nematicide furadan (100% mortality at 1 mg/ml concentration after 24 h).

  5. Genetic structure of Plasmodium vivax isolates from two malaria endemic areas in Afghanistan.

    PubMed

    Zakeri, Sedigheh; Safi, Najibullah; Afsharpad, Mandana; Butt, Waqar; Ghasemi, Faezeh; Mehrizi, Akram Abouie; Atta, Hoda; Zamani, Ghasem; Djadid, Navid Dinparast

    2010-01-01

    In this study, the nature and extent of genetic diversity of Plasmodium vivax populations circulating in Afghanistan have been investigated by analyzing three genetic markers: csp, msp-1, and msp-3 alpha. Blood samples (n=202) were collected from patients presenting with vivax malaria from south-western (Herat) and south-eastern (Nangarhar) parts of Afghanistan, and analysed using nested-PCR/RFLP and sequencing methods. Genotyping pvmsp-1 revealed type 1, type 2 and recombinant type 3 allelic variants, with type 1 predominant in parasites in both study areas. The sequence analysis of 57 P. vivax isolates identified a total of 26 distinct alleles. Genotyping pvcsp gene showed that VK210 type (86.6%) is predominant in Afghanistan. Moreover, three major types of the pvmsp-3 alpha locus: type A, type B and type C were distinguished among Afghani isolates. The predominant fragments among Nangarhar and Herat parasites were type A (70.8% and 67.9%, respectively). PCR/RFLP products with Hha I and Alu I were detected 52 and 38 distinct variants among Nangarhar and Herat isolates, respectively. These results strongly indicate that the P. vivax populations in Afghanistan are highly diverse.

  6. Lake char (Salvelinus namaycush) olfactory neurons are highly sensitive and specific to bile acids.

    PubMed

    Zhang, Chunbo; Hara, Toshiaki J

    2009-02-01

    Bile acids have been implicated as chemical signals in spawning behaviour of lake char (Salvelinus namaycush). In this study, we investigated olfactory responses of lake char to bile acids by using the electro-olfactogram recording. Lake char detected 9 out of 38 bile acids tested at thresholds 0.02-0.5 nM. The most stimulatory included chenodeoxycholic acid, cholic acid, taurochenodeoxycholic acid, taurocholic acid, and taurolithocholic acid 3alpha-sulphate. Structure-activity analysis indicated that substituents in the side chain or hydroxyl sulphation were determinant elements for the recognition of individual bile acid receptors, while the position and orientation of hydroxyls or the type of amidation were important for effective stimulation. Three distinct types of concentration-response relationships were found, representing free, taurine- or glycine-amidated, and 3alpha-sulphated bile acids. Cross-adaptation and binary mixture experiments revealed the presence of multiple olfactory receptors for bile acids. Lake char were also capable of detecting petromyzonol sulphate at 1 nM, possibly via its own receptors. Our study further showed that the olfactory responses to bile acids were independent of those of known odorants including amino acids, prostaglandins and gonadal steroids. We conclude that lake char possess multiple olfactory receptors capable of discriminating bile acids produced and released by conspecifics.

  7. Exercise regulates Akt and glycogen synthase kinase-3 activities in human skeletal muscle.

    PubMed

    Sakamoto, Kei; Arnolds, David E W; Ekberg, Ingvar; Thorell, Anders; Goodyear, Laurie J

    2004-06-25

    Activation of Akt and deactivation of GSK3 are critical signals regulating a number of cellular processes in multiple systems. Whether physical exercise alters Akt and GSK3 activity in human skeletal muscle is controversial. beta-Catenin, a GSK3 substrate and important Wnt signaling protein that alters gene transcription, has not been investigated in human skeletal muscle. In the present study, eight healthy human subjects performed 30min of cycling exercise at 75% of maximum workload (submaximal) followed by 6 bouts of 60s at 125% maximum workload (maximal). Biopsies of vastus lateralis muscle were taken at rest (basal), and within 15s following cessation of the submaximal and maximal exercise bouts. Exercise at both submaximal and maximal intensities significantly increased Akt activity (40% and 110%, respectively). Increases in Akt activity were accompanied by increases in Akt Thr(308) and Ser(473) phosphorylation, decreased GSK3alpha activity ( approximately 30% at both intensities), and increased phosphorylation of GSK3alpha Ser(21). Exercise at both intensities also decreased beta-catenin Ser(33/37)Thr(41) phosphorylation (50-60% at both intensities). These results demonstrate that Akt, GSK3, and beta-catenin signaling are regulated by exercise in human skeletal muscle, and as such identify them as possible molecular mediators of exercise's effect on metabolic and transcriptional processes in skeletal muscle.

  8. Bisdesmosidic saponins from Securidaca longepedunculata roots: evaluation of deterrency and toxicity to Coleopteran storage pests.

    PubMed

    Stevenson, Philip C; Dayarathna, Thamara K; Belmain, Steven R; Veitch, Nigel C

    2009-10-14

    Powdered dry root bark of Securidaca longepedunculata was mixed with maize and cowpea and effectively reduced the numbers of Sitophilus zeamais and Callosobruchus maculatus emerging from these commodities, respectively, more than 9 months after treatment. This effect was reciprocated in grain treated with a methanol extract of the root bark, indicating that compounds were present that were oviposition deterrents or directly toxic to the adults or larvae. Two new bisdesmosidic saponins, 3-O-beta-D-glucopyranosyl-28-O-(alpha-L-arabinopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)[beta-D-apiofuranosyl-(1 --> 3)]-alpha-L-rhamnopyranosyl-(1 --> 2)-[4-O-(4-methoxycinnamoyl-beta-D-fucopyranosyl)])-medicagenic acid (securidacaside A) and 3-O-beta-D-glucopyranosyl-28-O-(alpha-L-arabinopyranosyl-(1 --> 3)-beta-D-xylopyranosyl-(1 --> 4)[beta-D-apiofuranosyl-(1 --> 3)]-alpha-L-rhamnopyranosyl-(1 --> 2)-[4-O-(3,4,5-trimethoxy-(E)-cinnamoyl-beta-D-fucopyranosyl)])-medicagenic acid (securidacaside B), were isolated from the methanol extract of the roots of S. longepedunculata and characterized by spectroscopic methods. Securidacaside A, which occurred as (E)- and (Z)-regioisomers, showed deterrency and toxicity toward C. maculatus and S. zeamais and could contribute to the biological activity of the methanol extract. The potential to optimize the use of this plant for stored product protection using water extracts, which would be appropriate technology for target farmers, is discussed.

  9. The structure of the O-antigen in the endotoxin of the emerging food pathogen Cronobacter (Enterobacter) muytjensii strain 3270.

    PubMed

    MacLean, Leann L; Pagotto, Franco; Farber, Jeffrey M; Perry, Malcolm B

    2009-03-31

    Strains of the Gram-negative bacterium Cronobacter (formerly known as Enterobacter) sakazakii have been identified as emerging opportunistic pathogens that can cause enterocolitis, bacteraemia, meningitis, and brain abscess, and they have been particularly associated with meningitis in neonates where infant milk formulae have been epidemiologically linked to the disease. A study of the lipopolysaccharides produced by clinical isolates using chemical, 2D 1H and 13C NMR, and MS methods revealed that the O-polysaccharide produced by Cronobacter muytjensii strain 3270, isolated from powdered infant formula from Denmark, was a linear unbranched polymer of a repeating pentasaccharide unit composed of 2-acetamido-2-deoxy-d-galactose (d-GalNAc), 2-acetamido-2-deoxy-d-glucose (d-GlcNAc), 3-acetamido-3-deoxy-d-quinovose (d-Qui3NAc), l-rhamnose (l-Rha), and d-glucuronic acid (d-GlcA) in equimolar ratio, and has the structure -->3)-alpha-D-GalpNAc-(1-->4)-alpha-D-Quip3NAc-(1-->3)-alpha-L-Rhap-(1-->6)-alpha-D-GlcpNAc-(1-->4)-beta-D-GlcpA-(1--> The specific structural characteristics of the O-polysaccharides of C. muytjensii may be of value in the identification and tracking of the bacterial pathogen.

  10. Disparate Degrees of Hypervariable Loop Flexibility Control T-Cell Receptor Cross-Reactivity, Specificity, and Binding Mechanism

    SciTech Connect

    Scott, Daniel R.; Borbulevych, Oleg Y.; Piepenbrink, Kurt H.; Corcelli, Steven A.; Baker, Brian M.

    2012-06-19

    {alpha}{beta} T-cell receptors (TCRs) recognize multiple antigenic peptides bound and presented by major histocompatibility complex molecules. TCR cross-reactivity has been attributed in part to the flexibility of TCR complementarity-determining region (CDR) loops, yet there have been limited direct studies of loop dynamics to determine the extent of its role. Here we studied the flexibility of the binding loops of the {alpha}{beta} TCR A6 using crystallographic, spectroscopic, and computational methods. A significant role for flexibility in binding and cross-reactivity was indicated only for the CDR3{alpha} and CDR3{beta} hypervariable loops. Examination of the energy landscapes of these two loops indicated that CDR3{beta} possesses a broad, smooth energy landscape, leading to rapid sampling in the free TCR of a range of conformations compatible with different ligands. The landscape for CDR3{alpha} is more rugged, resulting in more limited conformational sampling that leads to specificity for a reduced set of peptides as well as the major histocompatibility complex protein. In addition to informing on the mechanisms of cross-reactivity and specificity, the energy landscapes of the two loops indicate a complex mechanism for TCR binding, incorporating elements of both conformational selection and induced fit in a manner that blends features of popular models for TCR recognition.

  11. Bile acids. 38. Conversion of 5 -cholestane-3 ,7 -diol to allo bile acids by the rat.

    PubMed

    Noll, B W; Doisy, E A; Elliott, W H

    1973-07-01

    5alpha-[4-(14)C, 3alpha-(3)H]Cholestane-3beta,7alpha-diol was prepared from individual samples of 5alpha-[3alpha-(3)H]cholestane-3beta,7alpha-diol and 5alpha-[4-(14)C]cholestane-3beta,7alpha-diol, each derived from 3beta-acetoxycholest-5-en-7-one. Bile was collected for 11 days from adult male rats, with cannulated bile ducts, that had received intraperitoneally 0.90-0.92 mg of the doubly labeled diol. Bile from the first 10 hr, containing 63% of the administered (14)C and 6% of the (3)H, was hydrolyzed, and the bile acids were separated by acetic acid partition chromatography. Allochenodeoxycholic and allocholic acids contained at least 20.6% and 48.6%, respectively, of the (14)C retained in the biliary acids. Small amounts of (14)C (2.5% and 1.9%, respectively) were present in the 3beta isomers of these acids, but the tritium content totaled more than half of that found in the bile acid fraction. No evidence was obtained for presence of the extensive quantities of the allomuricholates.

  12. Acne and hirsutism in polycystic ovary syndrome: clinical, endocrine-metabolic and ultrasonographic differences.

    PubMed

    Falsetti, L; Gambera, A; Andrico, S; Sartori, E

    2002-08-01

    The aim of this study was to investigate whether the absence or presence of acne or hirsutism in 248 women with polycystic ovary syndrome was associated with different clinical, endocrine, metabolic and ultrasonographic factors. Patients were divided into three groups: 96 (38.7%) without any androgenic symptoms; 94 (37.9%) with only hirsutism; and 58 (23.4%) with only acne. The cycle alterations (oligomenorrhea or amenorrhea) and the echographic ovarian morphology (polycystic or multifollicular ovaries) showed no significant differences between the three groups. Hirsutism was associated with a greater incidence of obesity and insulin resistance, with an increase of 17-hydroxyprogesterone, ovarian and adrenal androgens, 3alpha-androstanediol glucuronide, insulin, insulin-like growth factor-I and low luteinizing hormone, sex hormone binding globulins and insulin-like growth factor binding protein-1 levels. Acne was associated only with the lowest 3alpha-androstanediol glucuronide levels. Therefore, two different pathogenetic mechanisms may play a role in the onset of acne and hirsutism.

  13. Progesterone metabolism in the pineal, brain stem, thalamus and corpus callosum of the female rat.

    PubMed

    Hanukoglu, I; Karavolas, H J; Goy, R W

    1977-04-15

    Specific brain regions, namely, thalamus, tectum, tegmentum, cerebellum, medulla and pineal, from five proestrous rats were incubated for 30 min with [3H]progesterone. After reverse isotopic dilution analysis, the following metabolites were identified in all incubations by purification to constant specific activity, derivative formation and/or gas liquid chromatography trapping: [3H]5alpha-pregnane-3, 20-dione (10-20% of the starting substrate except pineal -- 0.7%), [3H]3alpha-hydroxy-5alpha-pregnan-20-one (1.6-3.8% except for pineal -- 0.5%) and [3H]20alpha-hydroxy-4-pregnen-3-one (0.05-0.11%). Preliminary results from the corpus collosum incubation indicated the presence of the same metabolites. Although some apparent constant specific activities were obtained for 20alpha-hydroxy-5alpha-pregnan-3-one and 5beta-pregnane-3, 20-dione, the low levels of 3H associated with these steroids did not permit a definitive identification. The results indicate the presence of at least delta1-steroid 5alpha-reductase, 3alpha-hydroxysteroid dehydrogenase and 20alpha-hydroxysteroid dehydrogenase activities with progesterone as substrate in the brain regions examined.

  14. Biochemical determinants in Gender identity.

    PubMed

    Hamilton, W; Chapman, P H

    1977-01-01

    The purpose of this communication is to report cognate studies which suggest that the nature of the peripheral metabolism of testosterone may impart gender direction to thought construction and motive. In patients with the complete testicular feminizing syndrome [4], the XO/XY syndrome [4], female trans-sexualism [4] and testicular agenesis [5] HCG-tests of 3 days duration were performed, and plasma and urinary testosterone, urinary excretion of 5 alpha-androstane, 3 alpha, 17 beta-diol (5 alpha-diol), 5 beta-androstane, 3 alpha, 17 beta-diol (5 beta-diol) and epiandrosterone before and after stimulation were measured. In addition steroid transformation was examined by incubation studies with human fetal brain tissue. The results of the latter method presented here are in agreement with published work. It seems clear therefore that the peripheral levels of androgens, oestrogens and their metabolites combine with cerebral steroid transformation, metabolism and possible also synthesis in order to establish gender identity. Exploration of the role of peripheral hormones as stimulators of both gender identity and gender function has dictated the need for a new approach to therapy for gender abnormalities in psyche and soma.

  15. Characterization of flavonols in cranberry (Vaccinium macrocarpon) powder.

    PubMed

    Vvedenskaya, Irina O; Rosen, Robert T; Guido, Jane E; Russell, David J; Mills, Kent A; Vorsa, Nicholi

    2004-01-28

    Flavonoids were extracted from cranberry powder with acetone and ethyl acetate and subsequently fractionated with Sephadex LH-20 column chromatography. The fraction eluted with a 60% methanol solution was composed primarily of phenolic constituents with maximum absorbance at 340 nm. A high-performance liquid chromatography procedure was developed, which resolved 22 distinct peaks with UV/vis and mass spectra corresponding to flavonol glycoside conjugates. Six new constituents not previously reported in cranberry or in cranberry products were determined through NMR spectroscopy to be myricetin-3-beta-xylopyranoside, quercetin-3-beta-glucoside, quercetin-3-alpha-arabinopyranoside, 3'-methoxyquercetin-3-alpha-xylopyranoside, quercetin-3-O-(6' '-p-coumaroyl)-beta-galactoside, and quercetin-3-O-(6' '-benzoyl)-beta-galactoside. Quercetin-3-O-(6' '-p-coumaroyl)-beta-galactoside and quercetin-3-O-(6' '-benzoyl)-beta-galactoside represent a new class of cranberry flavonol compounds with three conjugated components consisting of a flavonol, sugar, and carboxylic acid (benzoic or hydroxycinnamic acids). This is also the first report identifying quercetin-3-arabinoside in both furanose and pyranose forms in cranberry. Elucidation of specific flavonol glycosides in cranberry is significant since the specificity of the sugar moiety may play a role in the bioavailability of the flavonol glycosides in vivo.

  16. The protein phosphatase-1/inhibitor-2 complex differentially regulates GSK3 dephosphorylation and increases sarcoplasmic/endoplasmic reticulum calcium ATPase 2 levels

    SciTech Connect

    King, Taj D.; Gandy, Johanna C.; Bijur, Gautam N. . E-mail: gautam@uab.edu

    2006-11-01

    The ubiquitously expressed protein glycogen synthase kinase-3 (GSK3) is constitutively active, however its activity is markedly diminished following phosphorylation of Ser21 of GSK3{alpha} and Ser9 of GSK3{beta}. Although several kinases are known to phosphorylate Ser21/9 of GSK3, for example Akt, relatively much less is known about the mechanisms that cause the dephosphorylation of GSK3 at Ser21/9. In the present study KCl-induced plasma membrane depolarization of SH-SY5Y cells, which increases intracellular calcium concentrations caused a transient decrease in the phosphorylation of Akt at Thr308 and Ser473, and GSK3 at Ser21/9. Overexpression of the selective protein phosphatase-1 inhibitor protein, inhibitor-2, increased basal GSK3 phosphorylation at Ser21/9 and significantly blocked the KCl-induced dephosphorylation of GSK3{beta}, but not GSK3{alpha}. The phosphorylation of Akt was not affected by the overexpression of inhibitor-2. GSK3 activity is known to affect sarcoplasmic/endoplasmic reticulum calcium ATPase 2 (SERCA2) levels. Overexpression of inhibitor-2 or treatment of cells with the GSK3 inhibitors lithium and SB216763 increased the levels of SERCA2. These results indicate that the protein phosphatase-1/inhibitor-2 complex differentially regulates GSK3 dephosphorylation induced by KCl and that GSK3 activity regulates SERCA2 levels.

  17. The enzymic and chemical synthesis of ursodeoxycholic and chenodeoxycholic acid from cholic acid.

    PubMed

    Sutherland, J D; Macdonald, I A; Forrest, T P

    1982-01-01

    Three approaches to the synthesis of ursodeoxycholic acid (UDC) from cholic acid have been investigated: (i) oxidation of cholic acid to 3 alpha, 7 alpha-dihydroxy-12 keto-5 beta-cholanoic acid (12K-CDC) with Clostridium group P 12 alpha-hydroxysteroid dehydrogenase (HSDH), isomerization of 12K-CDC to 3 alpha, 7 beta-dihydroxy-12 keto-5 beta-cholanoic acid (12K-UDC) with Clostridium absonum 7 alpha- and 7 beta-HSDH and reduction of 12K-UDC by Wolff-Kishner to UDC; (ii) isomerization of cholic acid to ursocholic acid (UC) by C. absonum 7 alpha- and 7 beta-HSDH, oxidation of UC to 12K-UDC with Clostridium group P 12 alpha-HSDH and Wolff-Kishner reduction of 12K-UDC to UDC; (iii) oxidation of cholic acid to 12K-CDC by Clostridium group P 12 alpha-HSDH, Wolff-Kishner reduction of 12K-CDC to chenodeoxycholic acid (CDC) and isomerization of CDC to UDC using whole cell cultures of C. absonum. In the first two approaches (using cell free systems) the yields of desired product were relatively low primarily due to the formation of various side products. The third method proved the most successful giving an overall yield of 37% (UDC) whose structure was verified by mass spectroscopy of the methyl ester.

  18. Combined chemical-enzymic synthesis of deoxygenated oligosaccharide analogs: transfer of deoxygenated D-GlcpNAc residues from their UDP-GlcpNAc derivatives using N-acetylglucosaminyltransferase I.

    PubMed

    Srivastava, G; Alton, G; Hindsgaul, O

    1990-10-25

    The 3''-, 4''-, and 6''-deoxy analogs of UDP-GlcpNAc have been synthesized chemically and found to act as donor-substrates for N-acetylglucosaminyltransferase-I (GnT-I) from human milk. Incubation of UDP-GlcpNAc and these deoxy analogs with GnT-I in the presence of alpha-D-Manp-(1----3)-[alpha-D-Manp-(1----6)]-beta-D-Manp -O(CH2)8COOMe gave beta-D-GlcpNAc-(1----2)-alpha-D-Manp-(1----3)-[alpha-D-Manp- (1----6)]- beta-D-Manp-O(CH2)8COOMe (6), and the deoxy analogs 12-14 where HO-3, HO-4, and HO-6, respectively, of the beta-D-GlcNAc residue were replaced by hydrogen. The tetrasaccharide glycosides 6 and 12-14 were characterized by 1H-n.m.r. spectroscopy and evaluated as acceptors for GnT-II, the next enzyme in the pathway of biosynthesis of Asn-linked oligosaccharides. Deoxygenation of the 3-position of the beta-D-GlcNAc residue of 6 completely abolished its acceptor activity, whereas removal of HO-4 or HO-6 caused only modest decreases in activity.

  19. The tumor microenvironment: possible role of integrins and the extracellular matrix in tumor biological behavior of intratubular germ cell neoplasia and testicular seminomas.

    PubMed Central

    Timmer, A.; Oosterhuis, J. W.; Schraffordt Koops, H.; Sleijfer, D. T.; Szabo, B. G.; Timens, W.

    1994-01-01

    In the present study, we examined the distribution of integrin subunits and extracellular matrix proteins in normal testis, intratubular germ cell neoplasia (ITGCN), and primary and metastatic seminomas. Compared to normal testis in ITGCN, Sertoli cells showed increased expression of alpha 3, alpha 6, and beta 1 integrin subunits. Malignant intratubular germ cells stained for alpha 3, alpha 6, and beta 1 integrin subunits. Progression of ITGCN to invasive seminoma was associated with loss of alpha 3 integrin subunit expression by tumor cells. Consequent to this loss, it can be speculated that the strong expression on ITGCN may be related to the noninvasive character of the lesion as is also known from other noninvasive tumors. All tumors showed a strong expression of alpha 6 and beta 1 integrin subunits. The alpha 5 integrin subunit was weakly expressed in primary seminomas in all stages. No differences were observed in integrin expression between primary and metastatic tumors. The distribution of extracellular matrix proteins was heterogeneous and revealed clear architectural differences between seminomas that may reflect different stages of tumor stroma formation. To our knowledge, the results presented in this study provide the first information on the possible role of tumor-extracellular matrix interactions in the biological behavior of ITGCN and testicular seminomas. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:8178927

  20. Characterization of integrin receptors in normal and neoplastic human brain.

    PubMed Central

    Paulus, W.; Baur, I.; Schuppan, D.; Roggendorf, W.

    1993-01-01

    We studied the immunohistochemical expression of integrin alpha and beta chains in the normal and neoplastic human brain. Normal astrocytes expressed alpha 2, alpha 3, alpha 6, beta 1, and beta 4 chains in some areas facing major interstitial tissues, but they were consistently negative for the other integrins examined (alpha 4, alpha 5, alpha V, alpha L, alpha M, alpha X, beta 2, beta 3). Neoplastic astrocytes in vivo and in vitro showed increased expression of alpha 3 and beta 1, and some also of alpha 5, alpha V, beta 3, and beta 4. Neoexpression of alpha 4 and reduced levels of beta 4 were detected in glioblastoma vascular proliferations compared with normal endothelial cells. Oligodendroglioma, ependymoma, choroid plexus papilloma, pituitary adenoma, and meningioma cells showed the same integrin pattern as their normal counterparts. Adhesion assays using the astrocytoma cell lines U-138 MG and U-373 MG revealed strong attachment to collagen types I to VI and undulin, which was inhibited by antibodies to beta 1, but not by those to alpha 2, alpha 3, alpha 6, and alpha V. We conclude that astrocytomas show increased levels or neoexpression of various integrins and strong attachment to various extracellular matrix components, which appears to be almost exclusively mediated by beta 1-integrins. Images Figure 1 PMID:8317546

  1. [Biotransformation of artemisinic acid by cell suspension cultures of Cephalotaxus fortunei and Artemisia annua].

    PubMed

    Hu, Yan-shan; Zhu, Jian-hua; Jiang, Bo; Yu, Rong-min

    2010-05-01

    To investigate the biotransformation of artemisinic acid by cell suspension cultures of Cephalotaxus fortunei and Artemisia annua. Artemisinic acid was added into to the media of the suspension cells of Cephalotaxus fortunei and Artemisia annua in their logarithmic growth phase. The biotransfromed product was detected with HPLC and isolated by silica gel column, Sephadex LH20 and ODS chromatography methods. The chemical structure of biotransformed product was elucidated on the basis of physical-chemical properties and spectroscopic data. Otherwise, the influence of co-cultured time on conversion ratio was investigated with HPLC. One biotransformed product, 3-alpha-hydroxyartemisinic acid, was obtained after two days of artemisinic acid administration to the suspension cells of Cephalotaxus fortunei and Artemisia annua. The optimal co-cultured time in suspension cells of Cephalotaxus fortunei was 2 days with the highest biotransformation rate of 8.42%, and in the case of Artemisia annua, it was 3 days and 3.95% respectively. It was the first time for the biotransformation of artemisinic acid to 3-alpha-hydroxyartemisinic acid by using cell suspension cultures of Cephalotaxus fortunei and Artemisia annua.

  2. Steroid-binding proteins in rabbit plasma: Separation of testosterone-binding globulin (TeBG) from corticosteroid-binding globulin (CBG), preliminary characterization of TeBG, and changes in TeBG concentration during sexual maturation.

    PubMed

    Danzo, B J; Eller, B C

    1975-05-01

    Two distinct steroid-binding proteins are present in rabbit plasma. One of the proteins (TeBG) binds [3-H]5 alpha-dihydrotestosterone (5 alpha DHT) and [3-H]testosterone. The affinity of this binding protein for 5 alpha DHT was 3-4 times greater than for testosterone. Binding of [3-H]5 alphaDHT could be inhibited by unlabeled 5 alpha DHT, testosterone, 5 alpha-androstan-3 alpha,17 beta-diol (3 alpha-diol), and 17 alpha-methyl- B-testosterone (skf) 7690). The relative affinity of the competitors was: 5 alpha DHT greater than 3 alpha-diol greater than testosterone greater than SKF 7690. The antiandrogens, cyproterone (1,2 alpha-methylene-6-chloro-pregn-4,6-diene-17 alpla-ol-3,20 dione), cyproterone-17-acetate, and 6 alpha-bromo-17 beta-hydroxy-17 alpha-methyl-4-oxa-5 alpha-androstan-3-ine (BOMT) were ineffective in competing for [3-H5d alpha DHT binding sites, as were 4-androstene-3, 17-dione, 17 beta-estradiol (E2), progesterone, and cortisol. The formation of the [3-H]5 alpha DHT-TeBG complex was extremely rapid; the binding reaction was essentially completed in 15 s. The complex dissociated rapidly in the presence of charcoal. The dissociation rate constant (Kdiss) was 0.157 min- minus 1 and the dissociation half-time t-1/2) was 4.5 min. In the presence of charcoal and unlabeled 5 alpha DHT the Kdiss was 0.268 min- minus 1 and the t=1/2 was 2.6 min. The sedimentation coefficient of TeBG was congruent to 4.6 S and its molecular weight, estimated by gel filtration on a calibrated Sephadex G-200 column, was congruent to 75,000. The concentration of TeBG in male rabbit plasma decreased with sexual maturation and was approximately three times higher in adult females than in adult males. The other protein (CBG) bound both [3-H]cortisol and [3-H]progesterone. Binding of these compounds could be inhibited by unlabeled cortisol and progesterone, but not by unlabeled 5 alpha DHT, testosterone, or E2. CBG had a sedimentation of congruent to 3.9 S and an apparent molecular

  3. Differential pattern of integrin receptor expression in differentiated and anaplastic thyroid cancer cell lines.

    PubMed

    Hoffmann, S; Maschuw, K; Hassan, I; Reckzeh, B; Wunderlich, A; Lingelbach, S; Zielke, A

    2005-09-01

    Adhesion of tumor cells to the extracellular matrix (ECM) is a crucial step for the development of metastatic disease and is mediated by specific integrin receptor molecules (IRM). The pattern of metastatic spread differs substantially among the various histotypes of thyroid cancer (TC). However, IRM have only occasionally been characterized in TC until now. IRM expression was investigated in 10 differentiated (FTC133, 236, 238, HTC, HTC TSHr, XTC, PTC4.0/4.2, TPC1, Kat5) and two anaplastic TC cell lines (ATC, C643, Hth74), primary cultures of normal thyroid tissue (Thy1,3), and thyroid cancer specimens (TCS). Expression of 16 IRM (beta1-4, beta7, alpha1-6, alphaV, alphaIIb, alphaL, alphaM, alphaX) and of four IRM heterodimers (alpha2beta1, alpha5beta1, alphaVbeta3, alphaVbeta5), was analyzed by fluorescent-activated cell sorter (FACS) and immunohistochemical staining. Thyroid tumor cell adhesion to ECM proteins and their IRM expression in response to thyrotropin (TSH) was assessed. Follicular TC cell lines presented high levels of integrins alpha2, alpha3, alpha5, beta1, beta3 and low levels of alpha1, whereas papillary lines expressed a heterogenous pattern of IRM, dominated by alpha5 and beta1. ATC mainly displayed integrins alpha2, alpha3, alpha5, alpha6, beta1 and low levels of alpha1, alpha4 and alphaV. Integrin heterodimers correlated with monomer expression. Evaluation of TCS largely confirmed these results with few exceptions, namely alpha4, alpha6, and beta3. The ability of TC cell lines to adhere to purified ECM proteins correlated with IRM expression. TSH induced TC cell adhesion in a dose-dependent fashion, despite an unchanged array of IRM expression or level of a particular IRM. Thyroid carcinoma cell lines of different histogenetic background display profoundly different patterns of IRM expression that appear to correlate with tumor aggressiveness. In vitro adhesion to ECM proteins and IRM expression concur. Finally, TSH-stimulated adhesion of

  4. Expression of 10 GABA(A) receptor subunit messenger RNAs in the motor-related thalamic nuclei and basal ganglia of Macaca mulatta studied with in situ hybridization histochemistry.

    PubMed

    Kultas-Ilinsky, K; Leontiev, V; Whiting, P J

    1998-07-01

    In situ hybridization histochemistry technique with [35S]UTP-labelled riboprobes was used to study the expression pattern of 10 GABA(A) receptor subunit messenger RNAs in the basal ganglia and motor thalamic nuclei of rhesus monkey. Human transcripts were used for the synthesis of alpha2, alpha4, beta2, beta3, gamma1 and delta subunit messenger RNA probes. Rat complementary DNAs were used for generating alpha1, alpha3, beta1 and gamma2 subunit messenger RNA probes. Nigral, pallidal and cerebellar afferent territories in the ventral tier thalamic nuclei all expressed alpha1, alpha2, alpha3, alpha4, beta1, beta2, beta3, delta and gamma2 subunit messenger RNAs but at different levels. Each intralaminar nucleus displayed its own unique expression pattern. In the thalamus, gamma1 subunit messenger RNA was detected only in the parafascicular nucleus. Comparison of the expression patterns with the known organization of GABA(A) connections in thalamic nuclei suggests that (i) the composition of the receptor associated with reticulothalamic synapses, except for those in the intralaminar nuclei, may be alpha1alpha4beta2delta, (ii) receptors of various other subunit compositions may operate in the local GABAergic circuits, and (iii) the composition of receptors at nigro- and pallidothalamic synapses may differ, with those at nigrothalamic probably containing beta1 and gamma2 subunits. In the medial and lateral parts of the globus pallidus, the subthalamic nucleus and the substantia nigra pars reticularis, the alpha1, beta2 and gamma2 messenger RNAs were co-expressed at a high level suggesting that this subunit composition was associated with all GABAergic synapses in the direct and indirect striatal output pathways. Various other subunit messenger RNAs were also expressed but at a lower level. In the substantia nigra pars compacta the most highly expressed messenger RNAs were alpha3, alpha4 and beta3; all other subunit messenger RNAs studied, except for gamma1, alpha1 and

  5. Human liver class I alcohol dehydrogenase gammagamma isozyme: the sole cytosolic 3beta-hydroxysteroid dehydrogenase of iso bile acids.

    PubMed

    Marschall, H U; Oppermann, U C; Svensson, S; Nordling, E; Persson, B; Höög, J O; Jörnvall, H

    2000-04-01

    3beta-Hydroxy (iso) bile acids are formed during enterohepatic circulation from 3alpha-hydroxy bile acids and constitute normal compounds in plasma but are virtually absent in bile. Isoursodeoxycholic acid (isoUDCA) is a major metabolite of UDCA. In a recent study it was found that after administration of isoUDCA, UDCA became the major acid in bile. Thus, epimerization of the 3beta-hydroxy to a 3alpha-hydroxy group, catalyzed by 3beta-hydroxysteroid dehydrogenases (HSD) and 3-oxo-reductases must occur. The present study aims to characterize the human liver bile acid 3beta-HSD. Human liver cytosol and recombinant alcohol dehydrogenase (ADH) betabeta and gammagamma isozymes were subjected to native polyacrylamide gel electrophoresis (PAGE) and isoelectric focusing. Activity staining with oxidized nicotinamide adenine dinucleotide (NAD(+)) or oxidized nicotinamide adenine dinucleotide phosphate (NADP(+)) as cofactors and various iso bile acids as substrates was used to screen for 3beta-HSD activity. Reaction products were identified and quantified by gas chromotography/mass spectrometry (GC/MS). Computer-assisted substrate docking of isoUDCA to the active site of a 3-dimensional model of human class I gammagamma ADH was performed. ADH gammagamma isozyme was identified as the iso bile acid 3beta-HSD present in human liver cytosol, with NAD(+) as a cofactor. Values for k(cat)/K(m) were in the rank order isodeoxycholic acid (isoDCA), isochenodeoxycholic acid (isoCDCA), isoUDCA, and isolithocholic acid (isoLCA) (0.10, 0.09, 0.08, and 0. 05 min(-1) x micromol/L(-1), respectively). IsoUDCA fits as substrate to the 3-dimensional model of the active-site of ADH gammagamma. ADH gammagamma isozyme was defined as the only bile acid 3beta-HSD in human liver cytosol. Hydroxysteroid dehydrogenases are candidates for the binding and transport of 3alpha-hydroxy bile acids. We assume that ADH gammagamma isozyme is involved in cytosolic bile acid binding and transport processes as well.

  6. Formation of C21 bile acids from plant sterols in the rat

    SciTech Connect

    Boberg, K.M.; Lund, E.; Olund, J.; Bjoerkhem, I. )

    1990-05-15

    Formation of bile acids from sitosterol in bile-fistulated female Wistar rats was studied with use of 4-14C-labeled sitosterol and sitosterol labeled with 3H in specific positions. The major part (about 75%) of the 14C radioactivity recovered as bile acids in bile after intravenous administration of (4-14C)sitosterol was found to be considerably more polar than cholic acid, and only trace amounts of radioactivity had chromatographic properties similar to those of cholic acid and chenodeoxycholic acid. It was shown that polar metabolites were formed by intermediate oxidation of the 3 beta-hydroxyl group (loss of 3H from 3 alpha-3H-labeled sitosterol) and that the most polar fraction did not contain a hydroxyl group at C7 (retention of 3H in 7 alpha,7 beta-3H2-labeled sitosterol). Furthermore, the polar metabolites had lost at least the terminal 6 or 7 carbon atoms of the side chain (loss of 3H from 22,23-3H2- and 24,28-3H2-labeled sitosterol). Experiments with 3H-labeled 7 alpha-hydroxysitosterol and 4-14C-labeled 26-hydroxysitosterol showed that none of these compounds was an efficient precursor to the polar metabolites. By analysis of purified most polar products of (4-14C) sitosterol by radio-gas chromatography and the same products of 7 alpha,7 beta-(2H2)sitosterol by combined gas chromatography-mass spectrometry, two major metabolites could be identified as C21 bile acids. One metabolite had three hydroxyl groups (3 alpha, 15, and unknown), and one had two hydroxyl groups (3 alpha, 15) and one keto group. Considerably less C21 bile acids were formed from (4-14C)sitosterol in male than in female Wistar rats. The C21 bile acids formed in male rats did not contain a 15-hydroxyl group. Conversion of a (4-14C)sitosterol into C21 bile acids did also occur in adrenalectomized and ovariectomized rats, indicating that endocrine tissues are not involved.

  7. Antioxidant and free radical scavenging activity of flavonol glycosides from different Aconitum species.

    PubMed

    Braca, Alessandra; Fico, Gelsomina; Morelli, Ivano; De Simone, Francesco; Tomè, Franca; De Tommasi, Nunziatina

    2003-05-01

    Bioassay-guided fractionation by 1,1-diphenyl-2-dipicrylhydrazyl (DPPH) radical scavenging test of polar extracts of some Italian Aconitum species (A. napellus subsp. tauricum, A. napellus subsp. neomontanum, A. paniculatum, A. vulparia) led to the isolation of 13 flavonol glycosides: quercetin 3-O-(6-trans-caffeoyl)-beta-glucopyranosyl-(1-->2)-beta-glucopyranoside-7-O-alpha-rhamnopyranoside (1), kaempferol 3-O-(6-trans-caffeoyl)-beta-glucopyranosyl-(1-->2)-beta-glucopyranoside-7-O-alpha-rhamnopyranoside (2), quercetin 3-O-(6-trans-p-coumaroyl)-beta-glucopyranosyl-(1-->2)-beta-glucopyranoside-7-O-alpha-rhamnopyranoside (3), kaempferol 3-O-(6-trans-p-coumaroyl)-beta-glucopyranosyl-(1-->2)-beta-glucopyranoside-7-O-alpha-rhamnopyranoside (4), quercetin 7-O-(6-trans-caffeoyl)-beta-glucopyranosyl-(1-->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (5), kaempferol 7-O-(6-trans-caffeoyl)-beta-glucopyranosyl-(1-->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (6), kaempferol 7-O-(6-trans-p-coumaroyl)-beta-glucopyranosyl-(1-->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (7), kaempferol 3-O-beta-(2"-acetyl)galactopyranoside (8), kaempferol 3-O-beta-(2"-acetyl)galactopyranoside-7-O-alpha-arabinopyranoside (9), quercetin 3-O-beta-(2"-acetyl)galactopyranoside-7-O-alpha-arabinopyranoside (10), quercetin 3,7-di-O-alpha-rhamnopyranoside (11), kaempferol 3,7-di-O-alpha-rhamnopyranoside (12) and quercetin 3-O-beta-glucopyranoside-7-O-alpha-rhamnopyranoside (13). Their antioxidant activity (AA) was determined by measuring free radical scavenging activity by DPPH test and the coupled oxidation of beta-carotene and linoleic acid assay. The results showed that 5 is the most active compound in the DPPH free-radical scavenging test (IC(50) 1.9 microM) while in the coupled oxidation of beta-carotene and linoleic acid assay compound 1 has the highest inhibitory ratio after 1h (58.9%). Some structure-activity relationships on the AA were obtained.

  8. Exposure to inhalable dust, wheat flour and alpha-amylase allergens in industrial and traditional bakeries.

    PubMed

    Bulat, Petar; Myny, Katrien; Braeckman, Lutgart; van Sprundel, Marc; Kusters, Edouard; Doekes, Gert; Pössel, Kerstin; Droste, Jos; Vanhoorne, Michel

    2004-01-01

    This study was designed to characterize exposure to inhalable dust, wheat flour and alpha-amylase allergens in industrial and traditional bakeries. The study included 70 bakeries from the northern part of Belgium. Based on the degree of automation and a clear division of individual job tasks, four bakeries were identified as industrial and the remaining 66 were identified as traditional ones. Personal, as well as stationary, samples of inhalable dust were collected during full shift periods, usually 5-7 h. The portable pumps aspirated 2 l/min through Teflon personal dust samplers (Millipore, pore size 1.0 microm) mounted in PAS-6 sampling heads. In the collected samples the inhalable dust, wheat flour and alpha-amylase allergens were determined. Wheat flour allergens were measured using enzyme-linked immunosorbent assay inhibition and an antiwheat IgG4 serum pool. The alpha-amylase allergens were measured using a sandwich enzyme immunoassay with affinity-purified polyclonal rabbit IgG antibodies. In total, 440 samples (300 personal and 140 stationary) were processed. The highest inhalable dust exposure was observed in traditional bakeries among bread [geometric mean (GM) 2.10 mg/m3] and bread and pastry workers (GM 1.80 mg/m3). In industrial bakeries the highest dust exposure was measured in bread-producing workers (GM 1.06 mg/m3). Similar relations were observed for wheat flour and alpha-amylase allergens. Bread baking workers in traditional bakeries had the highest exposure to both allergens (wheat flour GM 22.33 microg/m(3), alpha-amylase GM 0.61 ng/m3). The exposure to wheat flour and alpha-amylase allergens in industrial bakeries was higher in bread baking workers (wheat flour GM 6.15 microg/m3, alpha-amylase GM 0.47 ng/m3) than in bread packing workers (wheat flour GM 2.79 microg/m3, alpha-amylase GM 0.15 ng/m3). The data presented suggest that, on average, exposure in the Belgium bakeries studied-industrial as well as traditional-is lower than or similar to

  9. [Biliary metabolites of levonorgestrel in rats].

    PubMed

    Minato, K; Takegawa, S; Koizumi, N; Tsukamoto, K; Honma, S

    1993-11-01

    The metabolism of levonorgestrel (LNG) in the bile following oral administration of the drug was examined in female rat. 1) Within 48 h after administration of 14C-labelled LNG (LNG-14C), 67-82% of the radioactivity was excreted into the bile. 2) Almost all the metabolites in the bile were conjugated with glucuronic acid or sulfuric acid and only a small amount of the unchanged compound was found. 3) After treatment of these metabolites in the bile with beta-glucuronidase and arylsulfatase, more than ten aglycones were detected on TLC. Three main aglycones, M1, M2 and M3, were isolated. They accounted for 68.0, 0.8 and 11.5% of the radioactivity excreted into the bile, respectively. 4) The structures of M1 and M2 were assumed to be 13-ethyl-18,19-dinor-5 alpha,17 beta-pregn-20-yne-3 alpha,17- diol and 13-ethyl-18,19-dinor-5 beta,17 beta-pregn-20-yne-3 alpha,17-diol, respectively, by NMR and LC/MS analyses, and confirmed by direct comparison with respective authentic samples. M3 was assigned to be 13-ethyl-18,19-dinor-5 alpha,17 beta-pregn-20-yne-3 alpha,16 beta,17-triol by NMR, LC/MS and GC/MS analyses and acetonide derivation. 5) Isolation of the glucuronide metabolite, M4, from the bile, was achieved by column chromatography using Amberlite XAD-2 and Sephadex LH-20. Hydrolysis of this compound with beta-glucuronidase released M1 and glucuronic acid. After M4 was converted to an acetylated-methyl ester derivative, the definite structural assignment of M4 was established to be M1-3-O-yl glucuronic acid by NMR analysis. The NOE effect and the value of the corresponding coupling constant of the anomeric proton showed that the glucoside moiety was in the beta configuration. These findings suggested that LNG was predominantly converted to 5 alpha-reduced metabolites and that the 5 beta-metabolite accounted for less than 1% of the total metabolites in female rats. These metabolites were excreted as glucuronides into the bile.

  10. Bioequivalence studies of tibolone in premenopausal women and effects on expression of the tibolone-metabolizing enzyme AKR1C (aldo-keto reductase) family caused by estradiol.

    PubMed

    Kang, Keon W; Kim, Yoon G

    2008-12-01

    This study aimed to investigate the bioequivalence of a test formulation of tibolone with the marketed reference formulation in 24 young healthy female volunteers. Tibolone is a synthetic steroid hormone for menopausal women. Volunteers were treated with the 2 formulations of tibolone (total dose of active ingredient 2.5 mg) according to a 2 x 2 crossover design with a 1-week washout period. Plasma concentrations of 3alpha- and 3beta-hydroxytibolone, which are major metabolites of tibolone, were assayed in timed samples over a 24-hour period with a validated gas chromatography/mass spectrometry (GC/MS) method that had a lower limit of quantification of 0.5 ng/mL. The reference and test formulations gave a mean 3alpha-hydroxytibolone C(max) of 5.0 and 5.2 ng/mL, respectively, and a mean 3beta-hydroxytibolone C(max) of 16.4 and 16.5 ng/mL, respectively. The mean AUC(t) of 3alpha-hydroxytibolone was 24.7 and 24.3 ng h/mL, whereas the mean AUC(t) of 3beta-hydroxytibolone was 57.6 and 54.8 ng h/mL for the test and reference formulations, respectively. The authors did not find significant differences in pharmacokinetic parameters between the 2 formulations, but metabolite formation was different from reports in postmenopausal women. The authors therefore measured the effects of estradiol on the expression of the tibolone-metabolizing enzymes, from the aldo-keto reductase (AKR1C) family, using HepG2 cell (human hepatoma cells) and MCF-7 cell (human breast cancer cells). Estradiol increased mRNA levels of AKR1C1, AKR1C2, and AKR1C3 and protein levels of total AKR1C in HepG2 cells. Estradiol selectively enhanced levels of AKR1C2 mRNA in MCF-7 cells. Thus, changes in the major metabolites of tibolone might result from changes in AKR1C family expression by patient estrogen status.

  11. Biocompatibility of solid-dosage forms of anti-human immunodeficiency virus type 1 microbicides with the human cervicovaginal mucosa modeled ex vivo.

    PubMed

    Trifonova, Radiana T; Pasicznyk, Jenna-Malia; Fichorova, Raina N

    2006-12-01

    Topical anti-human immunodeficiency virus (HIV) microbicides are being sought to reduce the spread of HIV type 1 (HIV-1) during sexual intercourse. The success of this strategy depends upon the selection of formulations compatible with the natural vaginal mucosal barrier. This study applied ex vivo-modeled human cervicovaginal epithelium to evaluate experimental solid-dosage forms of the anti-HIV-1 microbicide cellulose acetate 1,2-benzenedicarboxylate (CAP) and over-the-counter (OTC) vaginal products for their impact on inflammatory mediators regarded as potential HIV-1-enhancing risk factors. We assessed product-induced imbalances between interleukin-1alpha (IL-1alpha) and IL-1beta and the natural IL-1 receptor antagonist (IL-1RA) and changes in levels of IL-6, tumor necrosis factor alpha, IL-8, gamma interferon inducible protein 10 (IP-10), and macrophage inflammatory protein 3alpha (MIP-3alpha), known to recruit and activate monocytes, dendritic cells, and T cells to the inflamed mucosa. CAP film and gel formulation, similarly to the hydroxyethylcellulose universal vaginal placebo gel and the OTC K-Y moisturizing gel, were nontoxic and caused no significant changes in any inflammatory biomarker. In contrast, OTC vaginal cleansing and contraceptive films containing octoxynol-9 or nonoxynol-9 (N-9) demonstrated similar levels of toxicity but distinct immunoinflammatory profiles. IL-1alpha, IL-1beta, IL-8, and IP-10 were increased after treatment with both OTC vaginal cleansing and contraceptive films; however, MIP-3alpha was significantly elevated by the N-9-based film only (P < 0.01). Although both films increased extracellular IL-1RA, the cleansing film only significantly elevated the IL-1RA/IL-1 ratio (P < 0.001). The N-9-based film decreased intracellular IL-1RA (P < 0.05), which has anti-inflammatory intracrine functions. This study identifies immunoinflammatory biomarkers that can discriminate between formulations better than toxicity assays and should be

  12. Glycan flexibility: insights into nanosecond dynamics from a microsecond molecular dynamics simulation explaining an unusual nuclear Overhauser effect.

    PubMed

    Landström, Jens; Widmalm, Göran

    2010-01-26

    An atomistic all-atom molecular dynamics simulation of the trisaccharide beta-D-ManpNAc-(1-->4)[alpha-D-Glcp-(1-->3)]-alpha-L-Rhap-OMe with explicit solvent molecules has been carried out. The trisaccharide represents a model for the branching region of the O-chain polysaccharide of a strain from Aeromonas salmonicida. The extensive MD simulations having a 1-micros duration revealed a conformational dynamics process on the nanosecond time scale, that is, a 'time window' not extensively investigated for carbohydrates to date. The results obtained from the MD simulation underscore the predictive power of molecular simulations in studies of biomolecular systems and also explain an unusual nuclear Overhauser effect originating from conformational exchange. 2009 Elsevier Ltd. All rights reserved.

  13. Is there A Role for Alpha-Linolenic Acid in the Fetal Programming of Health?

    PubMed Central

    Leikin-Frenkel, Alicia I.

    2016-01-01

    The role of ω3 alpha linolenic acid (ALA) in the maternal diet during pregnancy and lactation, and its effect on the prevention of disease and programming of health in offspring, is largely unknown. Compared to ALA, ω3 docosahexaenoic (DHA) and eicosapentaenoic (EPA) acids have been more widely researched due to their direct implication in fetal neural development. In this literature search we found that ALA, the essential ω3 fatty acid and metabolic precursor of DHA and EPA has been, paradoxically, almost unexplored. In light of new and evolving findings, this review proposes that ALA may have an intrinsic role, beyond the role as metabolic parent of DHA and EPA, during fetal development as a regulator of gene programming for the prevention of metabolic disease and promotion of health in offspring. PMID:27023621

  14. KSC-2011-1903

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, space shuttle Endeavour approaches the Vehicle Assembly Building, or VAB, on its move from Orbiter Processing Facility-2 where it was processed for its final mission, STS-134. In the VAB, Endeavour will be lifted into a high bay where it will be joined to its external fuel tank and solid rocket boosters. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  15. KSC-2011-1793

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, space shuttle Endeavour approaches the Vehicle Assembly Building, or VAB, on its move from Orbiter Processing Facility-2 where it was processed for its final and upcoming STS-134 mission. In the VAB, Endeavour will be lifted into a high bay where it will be joined to its external fuel tank and solid rocket boosters. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  16. KSC-2011-1789

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, employees gather to hold up a banner to commemorate space shuttle Endeavour's STS-134 mission, as it is moved from Orbiter Processing Facility-2 to the Vehicle Assembly Building. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, spare parts, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  17. KSC-2011-1902

    NASA Image and Video Library

    2011-02-28

    CAPE CANAVERAL, Fla. -- At NASA's Kennedy Space Center in Florida, space shuttle Endeavour approaches the Vehicle Assembly Building, or VAB, on its move from Orbiter Processing Facility-2 where it was processed for its final mission, STS-134. In the VAB, Endeavour will be lifted into a high bay where it will be joined to its external fuel tank and solid rocket boosters. Endeavour and its STS-134 crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer, a high-pressure gas tank, additional spare parts for Dextre and micrometeoroid debris shields to the International Space Station. Launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Jack Pfaller

  18. Quantum Dot-Induced Phase Stabilization of ..alpha..-CsPbI3 Perovskite for High-Efficiency Photovoltaics

    SciTech Connect

    Swarnkar, Abhishek; Marshall, Ashley R.; Sanehira, Erin M.; Chernomordik, Boris D.; Moore, David T.; Christians, Jeffrey A.; Chakrabarti, Tamoghna; Luther, Joseph M.

    2016-10-07

    Here, we show nanoscale phase stabilization of CsPbI3 quantum dots (QDs) to low temperatures that can be used as the active component of efficient optoelectronic devices. CsPbI3 is an all-inorganic analog to the hybrid organic cation halide perovskites, but the cubic phase of bulk CsPbI3 (..alpha..-CsPbI3) -- the variant with desirable band gap -- is only stable at high temperatures. We also describe the formation of ..alpha..-CsPbI3 QD films that are phase-stable for months in ambient air. The films exhibit long-range electronic transport and were used to fabricate colloidal perovskite QD photovoltaic cells with an open-circuit voltage of 1.23 volts and efficiency of 10.77%. Furthermore, these devices function as light-emitting diodes with low turn-on voltage and tunable emission.

  19. [Antigenic bacterial polysaccharides. 23. The structure of the O-specific polysaccharide chain of Salmonella arizonae 059 lipopolysaccharide].

    PubMed

    Vinogradov, E V; Knirel', Iu A; Lipkind, G M; Shashkov, A S; Kochetkov, N K

    1987-09-01

    The O-specific polysaccharide of Salmonella arizonae O59 (Arizona 19) is composed of D-galactose, N-acetyl-D-glucosamine, and N-acetyl-L-fucosamine (FucNAc, 2-acetamido-2,6-dideoxy-L-galactose) in the ratio 1:1:1. The computerized calculation of the 13C NMR spectrum of the polysaccharide, based on the monosaccharide composition, spectra of the free monosaccharides and glycosydation effects, together with the chemical analysis (methylation and Smith degradation) showed that the polysaccharide is built up of trisaccharide repeating units of the following structure: ----3)-alpha-L-FucNAcp(1----3)-beta-D-GlcNAcp-(1----2)-beta- D-Galp-1(----. The molecular basis of serological interrelations between S. arizonae O59 and Pseudomonas aeruginosa O7 (Lányi) is discussed.

  20. Alpha Resonances in {sup 13}C Excited by the {sup 9}Be ({sup 6}Li,d) Reaction

    SciTech Connect

    Rodrigues, M. R. D.; Borello-Lewin, T.; Horodynski-Matsushigue, L. B.; Duarte, J. L. M.; Rodrigues, C. L.; Souza, M. A.; Miyake, H.; Cunsolo, A.; Cappuzzello, F.; Ukita, G. M.

    2010-05-21

    The {sup 9}Be({sup 6}Li,d){sup 13}C reaction was used to investigate alpha resonant states in {sup 13}C up to 13 MeV of excitation. The reaction was measured at a bombarding energy of 25.5 MeV employing the Sao Paulo Pelletron-Enge-Spectrograph facility and the nuclear emulsion detection technique. The resolution of 50 keV allowed for the separation of the resonant contributions to the known 7/2{sup -} at 10.753 MeV and (5/2{sup -}) at 10.818 MeV {sup 13}C states. The alpha resonance seen at the (3alpha+n) threshold was not previously reported. The experimental angular distributions are presented in comparison with DWBA predictions.

  1. Isolation and characterization of Exodus-2, a novel C-C chemokine with a unique 37-amino acid carboxyl-terminal extension.

    PubMed

    Hromas, R; Kim, C H; Klemsz, M; Krathwohl, M; Fife, K; Cooper, S; Schnizlein-Bick, C; Broxmeyer, H E

    1997-09-15

    Chemokines are a group of small, homologous proteins that regulate leukocyte migration, hemopoiesis, and HIV-1 absorption. We report here the cloning and characterization of a novel murine and human C-C chemokine termed Exodus-2 for its similarity to Exodus-1/MIP-3alpha/LARC, and its chemotactic ability. This novel chemokine has a unique 36 or 37 (murine and human, respectively) amino acid carboxyl-terminal extension not seen in any other chemokine family member. Purified recombinant Exodus-2 was found to have two activities classically associated with chemokines: inhibiting hemopoiesis and stimulating chemotaxis. However, Exodus-2 also had unusual characteristics for C-C chemokines. It selectively stimulated the chemotaxis of T-lymphocytes and was preferentially expressed in lymph node tissue. The combination of these characteristics may be a functional correlate for the unique carboxyl-terminal structure of Exodus-2.

  2. Anionic polymers of the cell wall of Brevibacterium linens VKM Ac-2159.

    PubMed

    Potekhina, N V; Shashkov, A S; Streshinskaya, G M; Senchenkova, S N; Evtushenko, L I

    2005-09-01

    Unsubstituted 1,3-poly(glycerol phosphate) and two sugar-1-phosphate polymers were identified in the cell wall of Brevibacterium linens VKM Ac-2159 by NMR spectroscopy and chemical methods. A monomer of one of the sugar-1-phosphate polymers has the branched repeating unit of the following structure: -4)-[beta-D-GlcpNAc-(1-->3)]-alpha-D-Glcp-(1-P-. The repeating unit of another sugar-1-phosphate polymer has a linear structure consisting of alternating beta- and alpha-N-acetylglucosamine residues: -4)-beta-D-GlcpNAc-(1-->6)-alpha-D-GlcpNAc-(1-P-. Some part of the beta-N-acetylglucosaminyl residues bear O-ester-bound succinic acid residues at C-3. The identified sugar-1-phosphate polymers have not been described earlier in cell walls of other bacteria.

  3. Quantum Dot-Induced Phase Stabilization of ..alpha..-CsPbI3 Perovskite for High-Efficiency Photovoltaics

    DOE PAGES

    Swarnkar, Abhishek; Marshall, Ashley R.; Sanehira, Erin M.; ...

    2016-10-07

    Here, we show nanoscale phase stabilization of CsPbI3 quantum dots (QDs) to low temperatures that can be used as the active component of efficient optoelectronic devices. CsPbI3 is an all-inorganic analog to the hybrid organic cation halide perovskites, but the cubic phase of bulk CsPbI3 (..alpha..-CsPbI3) -- the variant with desirable band gap -- is only stable at high temperatures. We also describe the formation of ..alpha..-CsPbI3 QD films that are phase-stable for months in ambient air. The films exhibit long-range electronic transport and were used to fabricate colloidal perovskite QD photovoltaic cells with an open-circuit voltage of 1.23 voltsmore » and efficiency of 10.77%. Furthermore, these devices function as light-emitting diodes with low turn-on voltage and tunable emission.« less

  4. Purification and characterization of a 4-hydroxybiphenyl UDP-glucuronosyltransferase from rat liver microsomes

    SciTech Connect

    Styczynski, B.; Green, M.; Coffman, B.; Puig, J.; Tephly, T. )

    1991-03-11

    A phenobarbital-inducible rat liver microsomal UDP-glucuronosyltransferase (4-HBP UDPGT) which catalyzes the glucuronidation of 4-hydroxybiphenyl has been purified to homogeneity. The apparent subunit molecular weight of this protein is 52,500 as determined by SDS-PAGE. 4-HBP UDPGT was shown to react with 4-hydroxybiphenyl, p-nitrophenol and 4-methylumbelliferone, but did not react with morphine, testosteron or chloramphenicol. Upon treatment with Endoglycosidase H, the 4-HBP UDPGT underwent about a 2,000 dalton decrease in subunit molecular weight, suggesting that this protein in N-glycosylated. Western blot analysis has revealed immunorecognition of 4-HBP UDPGT by antibodies raised in rabbit against rat 3{alpha}- and 17{beta}-hydroxysteroid UDPGTs. Additionally, the authors have obtained the N-terminal amino acid sequence of 4-HBP UDPGT. These data provide evidence which suggests that this protein is different from another UDPGT previously shown to react with 4-hydroxybiphenyl, testosterone and chloramphenicol.

  5. Rye polyphenols and the metabolism of n-3 fatty acids in rats: a dose dependent fatty fish-like effect

    PubMed Central

    Ounnas, Fayçal; de Lorgeril, Michel; Salen, Patricia; Laporte, François; Calani, Luca; Mena, Pedro; Brighenti, Furio; Del Rio, Daniele; Demeilliers, Christine

    2017-01-01

    As long-chain fatty acids (LCFA) of the n-3 series are critically important for human health, fish consumption has considerably increased in recent decades, resulting in overfishing to respond to the worldwide demand, to an extent that is not sustainable for consumers’ health, fisheries economy, and marine ecology. In a recent study, it has been shown that whole rye (WR) consumption improves blood and liver n-3 LCFA levels and gut microbiota composition in rats compared to refined rye. The present work demonstrates that specific colonic polyphenol metabolites may dose dependently stimulate the synthesis of n-3 LCFA, possibly through their microbial and hepatic metabolites in rats. The intake of plant n-3 alpha-linolenic acid and WR results in a sort of fatty fish-like effect, demonstrating that the n-3 LCFA levels in blood and tissues could be increased without eating marine foods, and therefore without promoting unsustainable overfishing, and without damaging marine ecology. PMID:28071699

  6. Rye polyphenols and the metabolism of n-3 fatty acids in rats: a dose dependent fatty fish-like effect.

    PubMed

    Ounnas, Fayçal; de Lorgeril, Michel; Salen, Patricia; Laporte, François; Calani, Luca; Mena, Pedro; Brighenti, Furio; Del Rio, Daniele; Demeilliers, Christine

    2017-01-10

    As long-chain fatty acids (LCFA) of the n-3 series are critically important for human health, fish consumption has considerably increased in recent decades, resulting in overfishing to respond to the worldwide demand, to an extent that is not sustainable for consumers' health, fisheries economy, and marine ecology. In a recent study, it has been shown that whole rye (WR) consumption improves blood and liver n-3 LCFA levels and gut microbiota composition in rats compared to refined rye. The present work demonstrates that specific colonic polyphenol metabolites may dose dependently stimulate the synthesis of n-3 LCFA, possibly through their microbial and hepatic metabolites in rats. The intake of plant n-3 alpha-linolenic acid and WR results in a sort of fatty fish-like effect, demonstrating that the n-3 LCFA levels in blood and tissues could be increased without eating marine foods, and therefore without promoting unsustainable overfishing, and without damaging marine ecology.

  7. Three-body model of light nuclei with microscopic nonlocal interactions

    SciTech Connect

    Theeten, M.; Baye, D.; Matsumura, H.; Orabi, M.; Descouvemont, P.; Fujiwara, Y.; Suzuki, Y.

    2007-11-15

    A three-body cluster model involving microscopic nonlocal interactions is developed and compared with a fully microscopic cluster model. The energy-independent nonlocal interactions are obtained from a renormalization of the energy-dependent kernels of the resonating-group method. Such interactions are derived for the {alpha}{alpha} and {alpha}n systems. The role and importance of nonlocality are discussed. These interactions are employed in three-body studies of the {alpha}nn,{alpha}{alpha}n, and 3{alpha} descriptions of the {sup 6}He, {sup 9}Be, and {sup 12}C nuclei. A comparison with fully microscopic calculations provides a measure of the importance of three-cluster exchanges in those states. The differences between both cluster-model calculations are in general small, except in the densities at short distances.

  8. Calculation of radiation therapy dose using all particle Monte Carlo transport

    DOEpatents

    Chandler, W.P.; Hartmann-Siantar, C.L.; Rathkopf, J.A.

    1999-02-09

    The actual radiation dose absorbed in the body is calculated using three-dimensional Monte Carlo transport. Neutrons, protons, deuterons, tritons, helium-3, alpha particles, photons, electrons, and positrons are transported in a completely coupled manner, using this Monte Carlo All-Particle Method (MCAPM). The major elements of the invention include: computer hardware, user description of the patient, description of the radiation source, physical databases, Monte Carlo transport, and output of dose distributions. This facilitated the estimation of dose distributions on a Cartesian grid for neutrons, photons, electrons, positrons, and heavy charged-particles incident on any biological target, with resolutions ranging from microns to centimeters. Calculations can be extended to estimate dose distributions on general-geometry (non-Cartesian) grids for biological and/or non-biological media. 57 figs.

  9. Calculation of radiation therapy dose using all particle Monte Carlo transport

    DOEpatents

    Chandler, William P.; Hartmann-Siantar, Christine L.; Rathkopf, James A.

    1999-01-01

    The actual radiation dose absorbed in the body is calculated using three-dimensional Monte Carlo transport. Neutrons, protons, deuterons, tritons, helium-3, alpha particles, photons, electrons, and positrons are transported in a completely coupled manner, using this Monte Carlo All-Particle Method (MCAPM). The major elements of the invention include: computer hardware, user description of the patient, description of the radiation source, physical databases, Monte Carlo transport, and output of dose distributions. This facilitated the estimation of dose distributions on a Cartesian grid for neutrons, photons, electrons, positrons, and heavy charged-particles incident on any biological target, with resolutions ranging from microns to centimeters. Calculations can be extended to estimate dose distributions on general-geometry (non-Cartesian) grids for biological and/or non-biological media.

  10. Evaluation of antinociceptive and anti-inflammatory activities of a new triterpene saponin from Bauhinia variegata leaves.

    PubMed

    Mohamed, Mona A; Mammoud, Madeha R; Hayen, Heiko

    2009-01-01

    A new triterpene saponin, named as 23-hydroxy-3alpha-[O-alpha-L-1C4-rhamnopyranosyl-(1"-->4')-O-alpha-L-4C1-arabinopyranosyl-oxy]olean-12-en-28-oic acid O-alpha-L-1C4-rhamnopyranosyl-(1"'-->4")-O-beta-D-4C1-glucopyranosyl-(1"-->6"')-O-beta-D-4C1-glucopyranosyl ester (9), was isolated from the leaves of Bauhinia variegata Linn. In addition, six flavonoid compounds along with two cinnamic acid derivatives were isolated and identified based on their chromatographic properties, and chemical and spectral data (ESI-high resolution-MSn, 1H NMR, 13C NMR, 1H-1H COSY, HSQC, and HMBC). Compound 9 was found to be nontoxic (LD50) and to have significant anti-inflammatory and antinociceptive activities. It also showed a slight antischistosomal activity.

  11. Systematic position of Histomonas meleagridis based on four protein genes.

    PubMed

    Hauck, Rüdiger; Hafez, Hafez M

    2010-04-01

    Phylogenetic trees based on parabasalid sequences of the small subunit rRNA placed Histomonas meleagridis in close proximity to Dientamoeba fragilis, Tritrichomonas foetus, and Monocercomonas sp. In this study, we sequenced partial genes of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), enolase, and alpha-tubulin from 2 strains of H. meleagridis. We found 5 different GAPDH sequences, 6 different enolase sequences, and 3 alpha-tubulin sequences. Phylogenetic trees based on the obtained sequences showed a close relationship of H. meleagridis with T. foetus and, to some extent, Monocercomonas sp. In conclusion, our findings further corroborate the ssu rRNA-based tree. Consequently, our study also supports the hypothesis that H. meleagridis secondarily lost cytoskeletal structures.

  12. Efficacy of methods for determining ovulation in a natural family planning program.

    PubMed

    Guida, M; Tommaselli, G A; Palomba, S; Pellicano, M; Moccia, G; Di Carlo, C; Nappi, C

    1999-11-01

    This methodological study aims to evaluate the efficacy of methods used for determining ovulation in a natural family planning in comparison with pelvic ultrasonography. Prospective analysis of ovulation detection methods was conducted with 40 women. Ovulation-detection methods employed in the study include transvaginal ultrasonography, daily morning urinary luteinizing hormone (LH) determinations, determination of salivary beta-glucuronidase activity, salivary ferning test, determination of cervical mucus levels and characteristics, and measurement of rectal or oral basal body temperature. Results revealed a 100% correlation between urinary LH level and ovulation diagnosis obtained from ultrasonographic examination. Other ovulation-detection methods contain the following correlation results: mucus sensations and characteristics, 48.3%; alpha-glucuronidase, 27.7%; and salivary ferning test, 36.8%. These findings draw the researchers to conclude that self-determination of LH levels could be an excellent method in ovulation determination.

  13. KSC-2011-1983

    NASA Image and Video Library

    2011-03-01

    CAPE CANAVERAL, Fla. -- A large yellow, metal sling lifts shuttle Endeavour from the transfer aisle into a high bay of the Vehicle Assembly Building at NASA's Kennedy Space Center in Florida. In the bay, the shuttle will be attached to its external fuel tank and solid rocket boosters. Endeavour is targeted to roll out to Kennedy's Launch Pad 39A for its final mission, STS-134, on March 9. Endeavour and the six-member crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank, additional spare parts for the Dextre robotic helper and micrometeoroid debris shields to the International Space Station. Endeavour's final launch is targeted for April 19 at 7:48 p.m. EDT. For more information visit, http://www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Frankie Martin

  14. Saponins from three species of Mimusops.

    PubMed

    Lavaud, C; Massiot, G; Becchi, M; Misra, G; Nigam, S K

    1996-02-01

    Six saponins were isolated from the seed kernel of Mimusops elengi, M. hexandra and M. manilkara. Their structures were determined using a combination of 1H NMR, 13C NMR and mass spectroscopy. Three of them are new compounds: 3-O-(beta-D-glucuronopyranosyl) 28-O-(alpha-L-rhamnopyranosyl (1-->3) beta-D-xylopyranosyl(1-->4) [alpha-L-rhamnopyranosyl(1-->3)] alpha-L-rhamnopyranosyl(1-->2) alpha-L-arabinopyranosyl) protobassic acid, 3-O-(beta-D-glucuronopyranosly) 28-O-(alpha-L-rhamnopyranosyl(1-->3) beta-D-xylopyranosyl(1-->4) alpha-L-rhamnopyranosyl(1-->2) alpha-L-arabinopyranosyl) 16-alpha-hydroxyprotobassic acid and 3-O-(beta-D-glucopyranosyl(1-->3) beta-D-glucopyranosyl) 28-O-(alpha-L-rhamnopyranosyl(1-->3) beta-D-xylopyranosyl(1-->4) alpha-L-rhamnopyranosyl(1-->2) alpha-L-arabinopyranosyl) protobassic acid.

  15. SDR-type human hydroxysteroid dehydrogenases involved in steroid hormone activation.

    PubMed

    Wu, Xiaoqiu; Lukacik, Petra; Kavanagh, Kathryn L; Oppermann, Udo

    2007-02-01

    Hydroxysteroid dehydrogenases catalyze the NAD(P)(H)-dependent oxidoreduction of hydroxyl and oxo-functions at distinct positions of steroid hormones. This reversible reaction constitutes an important pre-receptor control mechanism for nuclear receptor ligands of the androgen, estrogen and glucocorticoid classes, since the conversion "switches" between receptor ligands and their inactive metabolites. The major reversible activities found in mammals acting on steroid hormones comprise 3alpha-, 11beta- and 17beta-hydroxysteroid dehydrogenases, and for each group several distinct isozymes have been described. The enzymes differ in their expression pattern, nucleotide cofactor preference, steroid substrate specificity and subcellular localization, and thus constitute a complex system ensuring cell-specific adaptation and regulation of steroid hormone levels. Several isoforms constitute promising drug targets, of particular importance in cancer, metabolic diseases, neurodegeneration and immunity.

  16. Performance Prediction of the Mk II Navy 44 Sail Training Craft with respect to Tank Testing, Velocity Prediction Programs, and Computational Fluid Dynamics

    DTIC Science & Technology

    2003-05-05

    8217) end end Maple 6 Analysis Below is the script used to develop lift and drag for any yaw angle and rudder angle. > restart; > L:=(AH5, AH3 ...alpha1*AH5+alpha2)* AH3 ^2+(beta1*AH5+beta2)* AH3 +(gamma1*AH5+gam ma2); := L → ( ),AH5 AH3 + + + ( ) + α1 AH5 α2 AH32 ( ) + β1 AH5 β2 AH3 γ1 AH5 γ2...3 AG3 3 AG2 4 AG4 24 β1 4 AG5 AG6 AG7 = β1 − + − + + − 1 8 AG3 1 8 AG2 1 6 AG4 1 6 AG5 1 24 AG6 1 24 AG7 > L(AH5, AH3

  17. Evidence That GABA Mediates Dopaminergic and Serotonergic Pathways Associated with Locomotor Activity in Juvenile Chinook Salmon (Oncorhynchus tshawytscha)

    USGS Publications Warehouse

    Clements, S.; Schreck, C.B.

    2004-01-01

    The authors examined the control of locomotor activity in juvenile salmon (Oncorhynchus tshawytscha) by manipulating 3 neurotransmitter systems-gamma-amino-n-butyric acid (GABA), dopamine, and serotonin-as well as the neuropeptide corticotropin releasing hormone (CRH). Intracerebroventricular (ICV) injections of CRH and the GABAAagonist muscimol stimulated locomotor activity. The effect of muscimol was attenuated by administration of a dopamine receptor antagonist, haloperidol. Conversely, the administration of a dopamine uptake inhibitor (4???,4??? -difluoro-3-alpha-[diphenylmethoxy] tropane hydrochloride [DUI]) potentiated the effect of muscimol. They found no evidence that CRH-induced hyperactivity is mediated by dopaminergic systems following concurrent injections of haloperidol or DUI with CRH. Administration of muscimol either had no effect or attenuated the locomotor response to concurrent injections of CRH and fluoxetine, whereas the GABAA antagonist bicuculline methiodide potentiated the effect of CRH and fluoxetine.

  18. [Glycosides from flowers of Jasminum officinale L. var. grandiflorum].

    PubMed

    Zhao, Gui-qin; Xia, Jing-jing; Dong, Jun-xing

    2007-10-01

    To study the chemical constituents of the flower of Jasminum officinale L. var. grandiflorum. The compounds were isolated and purified by re-crystallization and chromatography on silica gel and Sephadex LH-20 column. Their structures were elucidated on the physicochemical properties and spectral analysis. Seven glycosides were identified as kaempferol-3-O-alpha-L-rhamnopyranosyl (1-->3)-[alpha-L-rhamnopyranosyl (1-->6)]-beta-D-galactopyranoside (I), kaempferol-3-O-rutinoside (II), 7-ketologanin (III), oleoside-11-methyl ester (IV), 7-glucosyl-l1-methyl oleoside (V), ligstroside (VI), oleuropein (VII). Compound I is a new compound. Compounds III and V were isolated from the family of Jasminum for the first time and compounds II, IV and VI were isolated from Jasminum officinale L. var. grandiflorum for the first time.

  19. Magnetic structures of actinide materials by pulsed neutron diffraction

    SciTech Connect

    Lawson, A.C.; Goldstone, J.A.; Huber, J.G.; Giorgi, A.L.; Conant, J.W.; Severing, A.; Cort, B.; Robinson, R.A.

    1990-01-01

    We describe some attempts to observe magnetic structure in various actinide (5f-electron) materials. Our experimental technique is neutron powder diffraction as practiced at a spallation (pulsed) neutron source. We will discuss our investigations of {alpha}-Pu, {delta}-Pu, {alpha}-UD{sub 3} and {beta}-UD{sub 3}. {beta}-UD{sub 3} is a simple ferromagnet: surprisingly, the moments on the two non-equivalent uranium atoms are the same within experimental error. {alpha}-UD{sub 3}, {alpha}-Pu and {delta}-Pu are non-magnetic, within the limits of our observations. Our work with pulsed neutron diffraction shows that it is a useful technique for research on magnetic materials.

  20. Studies on the formation of alpha-amylase by Thermomonospora vulgaris.

    PubMed

    Allam, A M; Hussein, A M; Ragab, A M

    1977-01-01

    Conditions affecting the formation of alpha-amylase by static cultures of the thermophilic actinomycete Thermomonospora vulgaris were studied. The organism failed to grow under submerged culture conditions or when the culture medium was devoid of CaCO3-alpha-Amylase was produced during the logarithmic phase of growth and maximum yield was obtained after 3 to 9 days of incubation. Growth and amylase formation took place only in a range from 45 degrees to 55 degrees C; optimum temperature was 55 degrees C. Of the tested carbon sources only starch induced enzyme formation. Maximum enzyme yield was obtained when starch concentration of the medium was 2% and when ammonium citrate served as a nitrogen source. Crushed clay pots could substitute for CaCO3 of the medium, but growth and amylase yield were less.

  1. KSC-2011-2701

    NASA Image and Video Library

    2011-04-01

    CAPE CANAVERAL, Fla. -- During a simulated pad emergency on Launch Pad 39A at NASA's Kennedy Space Center in Florida, members of the STS-134 crew check out a slidewire basket that would take them to a safe bunker below the pad in an unlikely emergency situation. The emergency training while aboard space shuttle Endeavour is part of a week-long Terminal Countdown Demonstration Test (TCDT). Endeavour's six crew members are targeted to launch April 19 at 7:48 p.m. EDT. They will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank, additional spare parts for the Dextre robotic helper and micrometeoroid debris shields to the International Space Station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  2. KSC-2011-2706

    NASA Image and Video Library

    2011-04-01

    CAPE CANAVERAL, Fla. - During a simulated pad emergency on Launch Pad 39A at NASA's Kennedy Space Center in Florida, STS-134 Mission Specialists Greg Chamitoff, left, and Andrew Feustel hop in a slidewire basket that would take them to a safe bunker below the pad in an unlikely emergency situation. The emergency training while aboard space shuttle Discovery is part of a week-long Terminal Countdown Demonstration Test (TCDT). Endeavour's six crew members are targeted to launch April 19 at 7:48 p.m. EDT. They will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank, additional spare parts for the Dextre robotic helper and micrometeoroid debris shields to the International Space Station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  3. KSC-2011-2704

    NASA Image and Video Library

    2011-04-01

    CAPE CANAVERAL, Fla. - During a simulated pad emergency on Launch Pad 39A at NASA's Kennedy Space Center in Florida, STS-134 Mission Specialists Andrew Feustel and Greg Chamitoff make their way toward a slidewire basket that would take them to a safe bunker below the pad in an unlikely emergency situation. The emergency training while aboard space shuttle Endeavour is part of a week-long Terminal Countdown Demonstration Test (TCDT). Endeavour's six crew members are targeted to launch April 19 at 7:48 p.m. EDT. They will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank, additional spare parts for the Dextre robotic helper and micrometeoroid debris shields to the International Space Station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  4. KSC-2011-2702

    NASA Image and Video Library

    2011-04-01

    CAPE CANAVERAL, Fla. - During a simulated pad emergency on Launch Pad 39A at NASA's Kennedy Space Center in Florida, STS-134 Mission Specialist Roberto Vittori, with the European Space Agency, hops in a slidewire basket that would take him to a safe bunker below the pad in an unlikely emergency situation. The emergency training while aboard space shuttle Discovery is part of a week-long Terminal Countdown Demonstration Test (TCDT). Endeavour's six crew members are targeted to launch April 19 at 7:48 p.m. EDT. They will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank, additional spare parts for the Dextre robotic helper and micrometeoroid debris shields to the International Space Station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  5. KSC-2011-2703

    NASA Image and Video Library

    2011-04-01

    CAPE CANAVERAL, Fla. -- During a simulated pad emergency on Launch Pad 39A at NASA's Kennedy Space Center in Florida, STS-134 Mission Specialist Andrew Feustel hops in a slidewire basket that would take him to a safe bunker below the pad in an unlikely emergency situation. The emergency training while aboard space shuttle Discovery is part of a week-long Terminal Countdown Demonstration Test (TCDT). Endeavour's six crew members are targeted to launch April 19 at 7:48 p.m. EDT. They will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank, additional spare parts for the Dextre robotic helper and micrometeoroid debris shields to the International Space Station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  6. KSC-2011-2700

    NASA Image and Video Library

    2011-04-01

    CAPE CANAVERAL, Fla. - During a simulated pad emergency on Launch Pad 39A at NASA's Kennedy Space Center in Florida, members of the STS-134 crew check out a slidewire basket that would take them to a safe bunker below the pad in an unlikely emergency situation. The emergency training while aboard space shuttle Endeavour is part of a week-long Terminal Countdown Demonstration Test (TCDT). Endeavour's six crew members are targeted to launch April 19 at 7:48 p.m. EDT. They will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank, additional spare parts for the Dextre robotic helper and micrometeoroid debris shields to the International Space Station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  7. Effects of (22S,23S)-Homobrassinolide and Related Compounds on Membrane Potential and Transport of Egeria Leaf Cells.

    PubMed

    Dahse, I; Sack, H; Bernstein, M; Petzold, U; Müller, E; Vorbrodt, H M; Adam, G

    1990-07-01

    (22S,23S)-Homobrassinolide was tested for its effect on the electric cell potential, proton extrusion, ferricyanide reduction, and amino acid and sucrose uptake of leaves of Egeria densa Planchon. In the light, (22S,23S)-homobrassinolide and its derivative, 2alpha-3alpha-dihydroxy-5alpha-stigmast-22-en-6-one, were similar to each other and similar to fusicoccin in causing hyperpolarization and proton extrusion, whereas stigmasterol was less effective. In darkness, the three sterols showed comparable effects. (22S,23S)-Homobrassinolide slightly stimulated ferricyanide reduction and promoted uptake of sucrose and alpha-aminoisobutyric acid. The results are compatible with a stimulation of an electrogenic proton pump mechanism at the plasmalemma by (22S,23S)-homobrassinolide.

  8. A new ferulic acid ester and other constituents from Tamarix nilotica leaves.

    PubMed

    Abouzid, Sameh Fekry; Ali, Sajjad Ahmed; Choudhary, Muhammad Iqbal

    2009-07-01

    Phytochemical investigation of the leaves of Tamarix nilotica (Tamaricaceae) has led to isolation of methyl ferulate 3-O-sulphate (1) for the first time from natural sources. In addition, coniferyl alcohol 4-O-sulphate (2), kaempferol 4'-methyl ether (3), tamarixetin (4) and quercetin 3-O-beta-D-glucupyranuronide (5) were isolated from the n-butanol soluble fraction of the extract. The pentacyclic triterpenoid, 3alpha-(3'',4''-dihydroxy-trans-cinnamoyloxy)-D-friedoolean-14-en-28-oic acid (6) was isolated from the n-hexane soluble fraction of the extract. The structures of these compounds were determined on the basis of spectroscopic analyses including 2 dimensional NMR. Compounds 3, 4 and 6 exhibited 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity with IC(50) values of 35.2, 37.0 and 21.2 muM, respectively.

  9. In vitro metabolism of androstenedione and identification of endogenous steroids in Helix aspersa

    SciTech Connect

    Le Guellec, D.; Thiard, M.C.; Remy-Martin, J.P.; Deray, A.; Gomot, L.; Adessi, G.L.

    1987-06-01

    In vitro metabolism of androstenedione in gonads of juvenile and adult Helix aspersa has been investigated. The conversion of (/sup 3/H)androstenedione into testosterone, 5 alpha-dihydrotestosterone, androsterone, and estriol was demonstrated. In juvenile animals testosterone (59.8%) is the major metabolite whereas in adult animals androsterone (18.8%) is. The following endogenous steroids have been identified by gas chromatography-mass spectrometry in adult gonads: androsterone, dehydroepiandrosterone, androstenedione, 3 alpha-androstanediol, estrone, estradiol-17 beta, and estriol. The levels of testosterone, 5 alpha-dihydrotestosterone, androstenedione, and progesterone have been measured by RIAs in gonads and hemolymph. Their levels vary with the physiological stage: the gonadal and circulating levels of testosterone decrease with the sexual maturation whereas the 5 alpha-dihydrotestosterone increases. These differences observed in metabolism and in level of steroids between the juvenile and the adult snails allow us to suppose that these steroids have a biological role.

  10. KSC-2011-3514

    NASA Image and Video Library

    2011-05-14

    CAPE CANAVERAL, Fla. -- In the Press Site auditorium at NASA's Kennedy Space Center in Florida, NASA managers brief media about their unanimous approval to proceed with space shuttle Endeavour's STS-134 mission. From left, are NASA Public Affairs Officer Candrea Thomas, Space Shuttle Program Launch Integration Manager Mike Moses, Shuttle Launch Director Mike Leinbach and Shuttle Weather Officer Kathy Winters. Endeavour's launch is scheduled for May 16 at 8:56 a.m. EDT. Endeavour and its crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Ben Smegelsky

  11. KSC-2011-3515

    NASA Image and Video Library

    2011-05-14

    CAPE CANAVERAL, Fla. -- In the Press Site auditorium at NASA's Kennedy Space Center in Florida, NASA managers brief media about their unanimous approval to proceed with space shuttle Endeavour's STS-134 mission. From left are Shuttle Weather Officer Kathy Winters, Shuttle Launch Director Mike Leinbach and Space Shuttle Program Launch Integration Manager Mike Moses. Endeavour's launch is scheduled for May 16 at 8:56 a.m. EDT. Endeavour and its crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Ben Smegelsky

  12. KSC-2011-3513

    NASA Image and Video Library

    2011-05-14

    CAPE CANAVERAL, Fla. -- In the Press Site auditorium at NASA's Kennedy Space Center in Florida, NASA managers brief media about their unanimous approval to proceed with space shuttle Endeavour's STS-134 mission. From left, are NASA Public Affairs Officer Candrea Thomas, Space Shuttle Program Launch Integration Manager Mike Moses, Shuttle Launch Director Mike Leinbach and Shuttle Weather Officer Kathy Winters. Endeavour's launch is scheduled for May 16 at 8:56 a.m. EDT. Endeavour and its crew will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the station. This will be the final spaceflight for Endeavour. For more information visit, www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Ben Smegelsky

  13. Intravenous infusion of iron and tetrahydrofolate does not influence intrauterine uteroferrin and secreted folate-binding protein content in swine.

    PubMed

    Vallet, J L; Christenson, R K; Klemcke, H G; Pearson, P L

    2001-01-01

    The effect of exogenous iron and folate on reproductive performance in swine is equivocal. However, the effect of exogenous iron and folate on secretion of their respective uterine transport proteins has never been reported. Twenty gilts were infused (n = 5 per treatment) with either 1) saline, 2) alpha-tocopherol, 3) alpha-tocopherol plus iron citrate, or 4) alpha-tocopherol plus tetrahydrofolate on d 11 to 14 of pregnancy. Intravenous infusion of iron citrate and tetrahydrofolate increased (P < 0.05) plasma iron and folate, respectively, for 6 to 8 h after treatment. Treatments had no effect on uterine content of uteroferrin or secreted folate-binding protein in uterine flushings obtained on d 15 of pregnancy. These data suggest that uterine secretion of uteroferrin and secreted folate-binding protein are not influenced by plasma levels of iron and folate, respectively, and may provide an explanation for the equivocal effect of iron and folate treatment on reproductive performance in gilts.

  14. Nonlinear optics of astaxanthin thin films

    NASA Astrophysics Data System (ADS)

    Esser, A.; Fisch, Herbert; Haas, Karl-Heinz; Haedicke, E.; Paust, J.; Schrof, Wolfgang; Ticktin, Anton

    1993-02-01

    Carotinoids exhibit large nonlinear optical properties due to their extended (pi) -electron system. Compared to other polyenes which show a broad distribution of conjugation lengths, carotinoids exhibit a well defined molecular structure, i.e. a well defined conjugation length. Therefore the carotinoid molecules can serve as model compounds to study the relationship between structure and nonlinear optical properties. In this paper the synthesis of four astaxanthins with C-numbers ranging from 30 to 60, their preparation into thin films, wavelength dispersive Third Harmonic Generation (THG) measurements and some molecular modelling calculations will be presented. Resonant (chi) (3) values reach 1.2(DOT)10-10 esu for C60 astaxanthin. In the nonresonant regime a figure of merit (chi) (3)/(alpha) of several 10-13 esu-cm is demonstrated.

  15. Modulation of the GABAA receptor by depressant barbiturates and pregnane steroids.

    PubMed Central

    Peters, J. A.; Kirkness, E. F.; Callachan, H.; Lambert, J. J.; Turner, A. J.

    1988-01-01

    1. The modulation of the gamma-aminobutyric acidA (GABAA) receptor by reduced metabolites of progesterone and deoxycorticosterone has been compared with that produced by depressant barbiturates in: (a) voltage-clamp recordings from bovine enzymatically isolated chromaffin cells in cell culture, and (b) an assay of the specific binding of [3H]-muscimol to a preparation of porcine brain membranes. 2. The progesterone metabolites 5 alpha- and 5 beta-pregnan-3 alpha-ol-20-one (greater than or equal to 30 nM) reversibly and dose-dependently enhanced the amplitude of membrane currents elicited by locally applied GABA (100 microM), and over the concentration range 30 nM-100 microM stimulated the binding of [3H]-muscimol. In contrast, 5 alpha- and 5 beta-pregnan-3 beta-ol-20-one (30 nM-100 microM) had little effect in either assay, indicating a marked stereoselectivity of steroid action. 3. Scatchard analysis of the ligand binding data suggested an apparent increase in the number, rather than the affinity, of detectable [3H]-muscimol binding sites as the principle action of the active steroid isomers. 4. GABA-evoked currents were also potentiated by androsterone (1 microM) and the deoxycorticosterone metabolite 5 alpha-pregnane-3 alpha,21-diol-20-one (100 nM). 5. Secobarbitone (10-100 microM), pentobarbitone (10-300 microM) and phenobarbitone (100-500 microM) reversibly and dose-dependently potentiated the amplitude of GABA-evoked currents in the absence of any change in their reversal potential. 6. At relatively high concentrations (greater than or equal to 30 microM) secobarbitone and pentobarbitone directly elicited a membrane current. It is concluded that such currents result from GABAA receptor-channel activation since they share a common reversal potential with GABA-evoked responses (approximately 0 mV), are reversibly antagonized by bicuculline (3 microM), and potentiated by either diazepam (1 microM) or 5 beta-pregnan-3 alpha-ol-20-one (500 nM). 7. Secobarbitone (1

  16. KSC-2011-3277

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At the NASA Shuttle Logistics Depot in Cape Canaveral, Florida, technicians remove the cover on the Load Control Assembly-2 (LCA-2) to begin the testing process. Located in space shuttle Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The LCA-2 will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  17. KSC-2011-3286

    NASA Image and Video Library

    2011-05-04

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, the Load Control Assembly-2 (LCA-2) has been replaced inside of space shuttle Endeavour. Located in Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission and has been replaced. Systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  18. KSC-2011-3285

    NASA Image and Video Library

    2011-05-04

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, the Load Control Assembly-2 (LCA-2) has been replaced inside of space shuttle Endeavour. Located in Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission and has been replaced. Systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  19. KSC-2011-3259

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, the Load Control Assembly-2 (LCA-2) is removed from inside space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The assembly will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  20. KSC-2011-3262

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. --The Load Control Assembly-2 (LCA-2) is outside at NASA Kennedy Space Center's Launch Pad 39A after it was removed from inside space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The assembly will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  1. KSC-2011-3281

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At the NASA Shuttle Logistics Depot in Cape Canaveral, Florida, the Load Control Assembly-2 (LCA-2) is uncovered for testing. Located in space shuttle Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The LCA-2 will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  2. KSC-2011-3276

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At the NASA Shuttle Logistics Depot in Cape Canaveral, Florida, technicians remove the cover on the Load Control Assembly-2 (LCA-2) to begin the testing process. Located in space shuttle Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The LCA-2 will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  3. KSC-2011-3284

    NASA Image and Video Library

    2011-05-04

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, the Load Control Assembly-2 (LCA-2) has been replaced inside of space shuttle Endeavour. Located in Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission and has been replaced. Systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  4. KSC-2011-3283

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At the NASA Shuttle Logistics Depot in Cape Canaveral, Florida, the Load Control Assembly-2 (LCA-2) is uncovered for testing. Located in space shuttle Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The LCA-2 will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  5. KSC-2011-3282

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At the NASA Shuttle Logistics Depot in Cape Canaveral, Florida, the Load Control Assembly-2 (LCA-2) is uncovered for testing. Located in space shuttle Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The LCA-2 will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  6. KSC-2011-3256

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, technicians work to remove the Load Control Assembly-2 (LCA-2) from inside space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The assembly will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  7. KSC-2011-3258

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, the Load Control Assembly-2 (LCA-2) is removed from inside space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The assembly will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  8. KSC-2011-3257

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, technicians begin to remove the Load Control Assembly-2 (LCA-2) from inside space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The assembly will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  9. KSC-2011-3275

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At the NASA Shuttle Logistics Depot in Cape Canaveral, Florida, technicians carefully remove the Load Control Assembly-2 (LCA-2) from a cart for testing. Located in space shuttle Endeavour's aft avionics bay 5, the LCA-2, which distributes power to nine shuttle systems, is believed to have caused fuel line heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The LCA-2 will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett

  10. Radical-scavenging activities of new hydroxylated ursane triterpenes from cv. Annurca apples.

    PubMed

    D'Abrosca, Brigida; Fiorentino, Antonio; Monaco, Pietro; Pacifico, Severina

    2005-07-01

    Two new ursolic acid triterpene derivatives, compounds 2 and 3, have been isolated from cv. Annurca apple fruit, a high-quality apple variety widely cultivated in southern Italy, together with the known 2-oxopomolic acid (1). The new compounds were identified by means of different spectroscopic techniques as 3-epi-2-oxopomolic acid (= (3alpha)-3,19-dihydroxy-2-oxours-12-en-28-oic acid; 2) and (1alpha)-1-hydroxy-3-oxours-12-en-28-oic acid (3). Compounds 1-3 were tested for their radical-scavenging activities with the aid of a 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay (Fig. 2). All three constituents showed activities similar to that of the reference antioxidant alpha-tocopherol (vitamin E).

  11. [Studies on the chemical constituents of Eclipta prostrata (L)].

    PubMed

    Zhang, J S; Guo, Q M

    2001-01-01

    To study the chemical constituents of Eclipta prostrata (L). The constituents of E. prostrata were systematically separated with the Bohlmann method and percolation and hot extraction methods, and various chromatographies. The structures were elucidated by chemical and spectroscopic means. Ten compounds were isolated from the Eclipta prostrata. Their structures were determined as wedelolactone (1), demethylwedelolactone (2), isodemethylwedelolactone (3), alpha-formylterthienyl (4), strychnolactone (5), beta-sitosterol (6), nonacosanol (7), stearic acid (8), lacceroic acid (9), 3,4-dihydoxy benzoic acid (10). Fourteen ocmpounds, including hydrocarbons and its esters were identified by GC-MS from the least polar fractions. Compound 3 is a new coumestan named isodemethylwedelolactone. Compounds 2-10 and compounds characterized by GC-MS analysis were obtained for the first time from Eclipta prostrata.

  12. Layered cathode materials for lithium ion rechargeable batteries

    DOEpatents

    Kang, Sun-Ho; Amine, Khalil

    2007-04-17

    A number of materials with the composition Li.sub.1+xNi.sub..alpha.Mn.sub..beta.Co.sub..gamma.M'.sub..delta.O.sub.2-- zF.sub.z (M'=Mg,Zn,Al,Ga,B,Zr,Ti) for use with rechargeable batteries, wherein x is between about 0 and 0.3, .alpha. is between about 0.2 and 0.6, .beta. is between about 0.2 and 0.6, .gamma. is between about 0 and 0.3, .delta. is between about 0 and 0.15, and z is between about 0 and 0.2. Adding the above metal and fluorine dopants affects capacity, impedance, and stability of the layered oxide structure during electrochemical cycling.

  13. A stereochemical examination of the equine metabolism of 17alpha-methyltestosterone.

    PubMed

    McKinney, Andrew R; Suann, Craig J; Stenhouse, Allen M

    2007-01-09

    An investigation was conducted into the stereochemistry of the equine urinary metabolites of 17alpha-methyltestosterone observed after oral administration. Standards of the complete range of C3/C5/C16 stereoisomeric 17alpha-methylandrostane-3,17beta-diols, 17alpha-methylandrostane-3,16,17beta-triols and 17alpha-hydroxymethylandrostane-3,17beta-diols were purchased or synthesised, and were used to unequivocally identify the absolute structures of the metabolites. Phase I metabolism was found to involve combinations of Delta(4)-3-ketone reduction with both 5alpha,3beta- and 5beta,3alpha-stereochemistry, hydroxylation at C16 with both 16alpha- and 16beta-stereochemistry and hydroxylation of the 17alpha-methyl substituent. Phase II metabolism involved mainly sulfation with a lesser degree of beta-glucuronidation.

  14. AKR1C2 — EDRN Public Portal

    Cancer.gov

    From NCBI Gene: This gene encodes a member of the aldo/keto reductase superfamily, which consists of more than 40 known enzymes and proteins. These enzymes catalyze the conversion of aldehydes and ketones to their corresponding alcohols using NADH and/or NADPH as cofactors. The enzymes display overlapping but distinct substrate specificity. This enzyme binds bile acid with high affinity, and shows minimal 3-alpha-hydroxysteroid dehydrogenase activity. This gene shares high sequence identity with three other gene members and is clustered with those three genes at chromosome 10p15-p14. Three transcript variants encoding two different isoforms have been found for this gene. [provided by RefSeq, Dec 2011

  15. Bile salts of the coelacanth, Latimeria chalumnae.

    PubMed

    Kihira, K; Akashi, Y; Kuroki, S; Yanagisawa, J; Nakayama, F; Hoshita, T

    1984-12-01

    Bile salts of the coelacanth, Latimeria chalumnae, Smith, have been analyzed and shown to have three bile alcohols, latimerol, 5 alpha-cyprinol, and 5 alpha-cholestane-3 beta, 7 alpha,-12 alpha,25,26-pentol, two C24 bile acids, chenodeoxycholic acid and cholic acid, one C26 bile acid, probably 3 beta, 7 alpha, 12 alpha-trihydroxy-27-nor-5 alpha-cholestan-26-oic acid, and two C27 bile acids, 3 alpha,7 alpha,12 alpha-trihydroxy-5 alpha-cholestan-26-oic acid and 3 beta,7 alpha,12 alpha-trihydroxy-5 alpha-cholestan-26-oic acid as determined by gas-liquid chromatography and gas-liquid chromatography-mass spectrometry.

  16. Sexual dimorphism involving steroidal pheromones and their binding protein in the submaxillary salivary gland of the Göttingen miniature pig.

    PubMed

    Booth, W D

    1984-02-01

    Submaxillary glands of mature Göttingen miniature pigs were examined for the presence of a sexual dimorphism. Gland weights, serous cell hypertrophy and total protein in the glands were much greater in male than female pigs. High concentrations of the pheromonal 16-androstene steroids were present in the glands of males and exceeded 2 mmol/g in some animals; this was primarily due to 5 alpha-androst-16-en-3 alpha-ol. The high concentration of 16-androstene steroids in boar glands was correlated with the presence of large amounts of binding protein for these steroids in the glands; smaller amounts of the binding protein were detected in female glands. These findings are similar to those found in domestic pigs, but the degree of sexual dimorphism assessed from these findings is more extreme in the miniature pig.

  17. Polysaccharides in Fungi. XXXIV. A polysaccharide from the fruiting bodies of Amanita muscaria and the antitumor activity of its carboxymethylated product.

    PubMed

    Kiho, T; Yoshida, I; Katsuragawa, M; Sakushima, M; Usui, S; Ukai, S

    1994-11-01

    A water-insoluble, alkali-soluble, glucan (AM-APP), [alpha]D +160 degrees in 0.4 M NaOH, was isolated from the alkaline extract of the fruiting bodies of Amanita muscaria. The results of chemical and spectroscopic investigations indicate that AM-APP is a linear (1 --> 3)-alpha-D-glucan with a molecular weigh estimated by gel chromatography of about 42000. Its carboxymethylated product (AM-APP-CM) showed potent antitumor activity against sarcoma 180 in mice, although the native polysaccharide (AM-APP) had little effect. The distribution of carboxymethyl groups in the molecule was analyzed by gas chromatography and mass spectrometry. The degree of substitution of carboxymethyl groups was 0.95 and the substituents were located at O-2, at O-4, at O-6, at O-2 and O-6, and at O-4 and O-6 on glucose.

  18. Genes expressed in the brain define three distinct neuronal nicotinic acetylcholine receptors.

    PubMed Central

    Nef, P; Oneyser, C; Alliod, C; Couturier, S; Ballivet, M

    1988-01-01

    Four genes encode the related protein subunits that assemble to form the nicotinic acetylcholine receptor (nAChR) at the motor endplate of vertebrates. We have isolated from the chicken genome four additional members of the same gene family whose protein products, termed alpha 2, alpha 3, alpha 4 and n alpha (non-alpha) probably define three distinct neuronal nAChR subtypes. The neuronal nAChR genes have identical structures consisting of six protein-coding exons and specify proteins that are best aligned with the chicken endplate alpha subunit, whose gene we have also characterized. mRNA transcripts encoding alpha 4 and n alpha are abundant in embryonic and in adult avian brain, whereas alpha 2 and alpha 3 transcripts are much scarcer. The same set of neuronal genes probably exists in all vertebrates since their counterparts have also been identified in the rat genome. Images PMID:3267226

  19. A highly regular fraction of a fucoidan from the brown seaweed Fucus distichus L.

    PubMed

    Bilan, Maria I; Grachev, Alexey A; Ustuzhanina, Nadezhda E; Shashkov, Alexander S; Nifantiev, Nikolay E; Usov, Anatolii I

    2004-02-25

    A fucoidan fraction consisting of L-fucose, sulfate, and acetate in a molar proportion of 1:1.21:0.08 was isolated from the brown seaweed Fucus distichus collected from the Barents Sea. The 13C NMR spectrum of the fraction was typical of regular polysaccharides containing disaccharide repeating units. According to 1D and 2D 1H and 13C NMR spectra, the fucoidan molecules are built up of alternating 3-linked alpha-L-fucopyranose 2,4-disulfate and 4-linked alpha-L-fucopyranose 2-sulfate residues: -->3)-alpha-L-Fucp-(2,4-di-SO3-)-(1-->4)-alpha-L-Fucp-(2SO3-)-(1-->. The regular structure may be only slightly masked by random acetylation and undersulfation of several disaccharide repeating units.

  20. Acylperoxylated and seco-mexicanolides from stems of Khaya anthotheca.

    PubMed

    Ferreira, Izabel C Piloto; Cortez, Diógenes A Garcia; das G F da Silva, M Fátima; Fo, Edson Rodrigues; Vieira, Paulo C; Fernandes, João B

    2005-03-01

    The stems of Khaya anthotheca yielded two new limonoids, which were identified on the basis of spectroscopic analysis as 1alpha,8alpha-oxido-3beta-acetoxy-2alpha-acylperoxy-1alpha,14alpha-dihydroxy-[3.3.1(10,2)]-bicyclomeliac-7,19-olide (3) and 3-acetoxy-8,14-dien-8,30-seco-khayalactone (4). Methyl 1alpha,2beta,3alpha,6,8alpha,14beta-hexahydroxy-[4.2.1(10,30).1(1,4)]-tricyclomeliac-7-oate, scopoletin, and 3-O-beta-D-glucopyranosylsitosterol were also isolated. The limonoids were of little value to clarify the basis of the nonresistance against Hypsipyla grandella.

  1. The chemical composition of R Coronae Borealis and XX Camelopardalis

    NASA Astrophysics Data System (ADS)

    Cottrell, P. L.; Lambert, D. L.

    1982-10-01

    The R Coronae Borealis stars R CrB and XX Cam are examined using extensive high-resolution Reticon data. Effective temperatures, values of log g, microturbulent velocities, and C/He values are determined, along with elemental abundances and abundance ratios for a wide range of species, including the light and heavy s-process elements. Helium- and carbon-rich models are employed, as is an appropriate model atmosphere code. The evolutionary history of R CrB stars is discussed in detail. It is concluded that both CNO cycle and 3-alpha processed material must have been mixed into the observable layers. The He core and He shell flashes are suggested as possible triggers for the mixing and for the extensive mass loss required to expose the heavily processed layers.

  2. Formation of a highly stable complex between BN 50730 [tetrahydro-4,7,8,10 methyl-1(chloro-2 phenyl)-6 (methoxy-4 phenyl-carbamoyl)-9 pyrido [4',3'-4,5] thieno [3,2-f] triazolo-1,2,4 [4,3-a] diazepine-1,4] and the platelet-activating factor receptor in rabbit platelet membranes.

    PubMed

    Silva, C L; Cruz, H N; Violante, F A; Cordeiro, R S; Martins, M A; Noël, F

    1996-01-26

    BN 50730 [tetrahydro-4,7,8,10 methyl-1(chloro-2 phenyl)-6 (methoxy-4 phenyl-carbamoyl)-9 pyrido [4',3'-4,5] thieno [3,2-f] triazolo-1,2,4 [4,3-alpha] diazepine-1,4], a novel platelet-activating factor (PAF) receptor antagonist with a hetrazepine structure, decreased the maximal number of binding sites (Bmax) of [3H]PAF in rabbit platelet membranes without altering its dissociation constant. Platelet aggregation induced by 1 microM PAF was prevented by preincubation with 1 microM BN 50730. The washing of the platelets preincubated with BN 50730 failed to revert its inhibitory effects. We conclude that BN 50730 acts as a non-competitive antagonist of the PAF receptor, due to the formation of a highly stable drug-receptor complex.

  3. Statistical methodology and assessment of seismic event characterization capability. Final report, 2 June 1993-2 September 1995

    SciTech Connect

    Fisk, M.D.; Gray, H.L.; McCartor, G.D.

    1995-10-31

    This project has focused on developing and applying statistical methods to perform seismic event characterization/identification and on quantifying capabilities with regard to monitoring of a Comprehensive Test Ban. An automated procedure is described to categorize seismic events, based on multivariate analysis of features derived from seismic waveforms. Second, preliminary event identification results are presented for a seismic event which occurred on 5 January 1995 in the Southern Ural Mountains region. Third, various statistics are compiled regarding 1786 seismic events which occurred between 11 January 1995 and 12 February 1995 and were detected by a set of 30 GSETT-3 Alpha stations. Fourth, a fundamental problem is addressed of how to utilize multivariate discriminant data from a multistation network in order to optimize the power of the outlier test for fixed false alarm rate.

  4. Duration of repellency of N,N-diethyl-3-methylbenzamide, citronella oil and cypermethrin against Culicoides species when applied to polyester mesh.

    PubMed

    Page, P C; Labuschagne, K; Nurton, J P; Venter, G J; Guthrie, A J

    2009-07-07

    The repellent efficacy of 15% N,N-diethyl-3-methylbenzamide (DEET), 0.6% citronella oil, and 0.3% alpha-cyano-cypermethrin against Culicoides species was compared in three 5x5 Latin squares (15 replicates) under South African field conditions. DEET, citronella oil or alpha-cyano-cypermethrin were applied to polyester meshes that were fitted to down-draught suction 220V UV light traps which were operated overnight. No significant repellent effect against Culicoides was found for the citronella oil or the alpha-cyano-cypermethrin. DEET had a significant repellent effect against Culicoides species and C. imicola for all catches made from after sunset to before sunrise.

  5. Targeting the BH3 domain of Bcl-2 family proteins. A brief history from natural products to foldamers as promising cancer therapeutic avenues.

    PubMed

    De Giorgi, M; Voisin-Chiret, A S; Rault, S

    2013-01-01

    For many years the spotlight in drug discovery has been on a relatively small number of validated therapeutic target classes, such as G-protein coupled receptors and enzymes such as protein kinases, with well characterized enzymatic and cellular activities. However, with recent progress in genomics and proteomics, protein-protein interactions (PPIs) provide new way of finding novel bioactive molecules acting on their interfaces. This review addresses the current case studies and state of the art in the development of small chemical modulators controlling interactions of proteins that have pathological implications in various human diseases and in particular in cancer. The attention is focused on Bcl-2 family protein modulators ranging from natural products to synthetic ones with particular interest in foldamers as BH3 alpha helix mimetics.

  6. Quantification of tocopherols and tocotrienols in portuguese olive oils using HPLC with three different detection systems.

    PubMed

    Cunha, Sara C; Amaral, Joana S; Fernandes, José O; Oliveira, M Beatriz P P

    2006-05-03

    Three different HPLC detection systems were compared for the determination of tocopherols and tocotrienols in olive oil: fluorescence and diode array connected in series, ultraviolet, and evaporative light scattering. The best results were obtained with the fluorescence detector, which was successfully applied in the quantification of tocopherols and tocotrienols in 18 samples of Portuguese olive oils. To support the validity of the method, the parameters evaluated were linearity, detection limits, repeatability, and recovery. All of the studied samples showed similar qualitative profiles with six identified compounds: alpha-T, beta-T, gamma-T, delta-T, alpha-T3, and gamma-T3. Alpha-tocopherol (alpha-T) was the main vitamin E isomer in all samples ranging from 93 to 260 mg/kg. The total tocopherols and tocotrienols ranged from 100 to 270 mg/kg. Geographic origin did not seem to influence the tocopherol and tocotrienol composition of the olive oils under evaluation.

  7. Constituents of Pterocarpus marsupium: an ayurvedic crude drug.

    PubMed

    Maurya, Rakesh; Singh, Rajinder; Deepak, Mundkinajeddu; Handa, S S; Yadav, Prem P; Mishra, Pushpesh K

    2004-04-01

    Five new flavonoid C-glucosides, 6-hydroxy-2-(4-hydroxybenzyl)-benzofuran-7-C-beta-d-glucopyranoside (1), 3-(alpha-methoxy-4-hydroxybenzylidene)-6-hydroxybenzo-2(3H)-furanone-7-C-beta-d-glucopyranoside (2), 2-hydroxy-2-p-hydroxybenzyl-3(2H)-6-hydroxybenzofuranone-7-C-beta-d-glucopyranoside (4), 8-(C-beta-d-glucopyranosyl)-7,3',4'-trihydroxyflavone (5) and 1,2-bis(2,4-dihydroxy,3-C-glucopyranosyl)-ethanedione (6) and two known compounds C-beta-d-glucopyranosyl-2,6-dihydroxyl benzene (7) and sesquiterpene (8), were isolated from an aqueous extract of the heartwood of Pterocarpus marsupium. The structure has been established using spectroscopic data.

  8. Histological study of the circulatory system of human dental pulp from individuals under local anesthesia and electro-acupuncture.

    PubMed

    Ueki, S; Iwai-Liao, Y; Han, K S; Higashi, Y

    1995-03-01

    A transmission electron microscopic (TEM) study was conducted on dental pulp obtained from patients under acupuncture or infiltration local analgesia. It was difficult to differentiate lymphatic circulation in the dental pulp that received infiltration anesthesia, because the vessels were constricted, congested, and showed stasis and thrombosis. On the other hand, the dental pulp that received acupuncture showed normal arterioles, capillaries, and venules, as well as some lymph capillaries and small efferent lymphatic vessels that measured about 8 microns and 100 microns in diameter, respectively. The lymphatic endothelial walls had many intercellular gaps, an imperfect basal lamina, and a few discontinuous pericytes. Between the openings in the lymphatic vessels, there were bundles of junctional filaments extending towards the dental pulp connective tissue. Therefore, the lymphatic system, which contains mainly B-3-alpha capillaries, is a leaky tissue for regulating fluid in the dental pulp.

  9. Mathematical Perspectives on the Federal Thrift Savings Plan (TSP)

    DTIC Science & Technology

    2008-06-10

    3 Alpha = 5 Alpha = 10 © 2008 Scott T. Nestler 38 Performance (Reward-Risk) Measures • Sharpe Ratio • STARR Ratio • R- Ratio ( Rachev ) ( ) ( ) E XSTARR...Similar to R- Ratio but use Alpha-VaR and Beta-VaR in place of CVaR • AVaR- Ratio : • BVaR- Ratio : 1 2 ( ) ( ) AVaR X AVR AVaR X α α − = 1 1 2 2...87,754 Beta VaR(50,5) $ 43,938 $ 44,010 $ 60,203 $ 67,757 Performance Measure (↑ better) VG-ICA TSP MP L 2030 L 2040 Sharpe Ratio 0.94 0.63 0.57

  10. Proteomic analysis of ethanol-induced embryotoxicity in cultured post-implantation rat embryos.

    PubMed

    Usami, Makoto; Mitsunaga, Katsuyoshi; Irie, Tomohiko; Miyajima, Atsuko; Doi, Osamu

    2014-04-01

    Protein expression changes were examined in day 10.5 rat embryos cultured for 24 hr in the presence of ethanol by using two-dimensional electrophoresis and mass spectrometry. Exposure to ethanol resulted in quantitative changes in many embryonic protein spots (16 decreased and 28 increased) at in vitro embryotoxic concentrations (130 and 195 mM); most changes occurred in a concentration-dependent manner. For these protein spots, 17 proteins were identified, including protein disulfide isomerase A3, alpha-fetoprotein, phosphorylated cofilin-1, and serum albumin. From the gene ontology classification and pathway mapping of the identified proteins, it was found that ethanol affected several biological processes involving oxidative stress and retinoid metabolism.

  11. Chemical composition and antimicrobial activity of the essential oil from Ambrosia trifida L.

    PubMed

    Wang, Peng; Kong, Chui Hua; Zhang, Chao Xian

    2006-07-25

    The essential oil obtained by steam distillation of dried aerial parts of Ambrosia trifida L. from Northeast China was analyzed by GC and GC-MS. The essential oil yield based on dried plant material was 0.12% and thirty-five compounds (corresponding to 86.7% of the total weight) were identified. The main components were: bornyl acetate (15.5%), borneol (8.5%), caryophyllene oxide (8.3%), alpha-pinene (8.0%), germacrene D (6.3%), beta-caryophyllene (4.6%), trans-carveol (2.9%), beta-myrcene (2.6%), camphor (2.4%) and limonene (3.2%). A. trifida essential oil demonstrated bactericidal and fungicidal activity against six bacterial strains and two fungal strains, using the agar diffusion method.

  12. Triumphalone, a diketone from the volatile oil of the leaves of Melaleuca triumphalis, and its spontaneous conversion into isotriumphalone.

    PubMed

    Brophy, Joseph J; Craig, Donald C; Goldsack, Robert J; Fookes, Christopher J R; Leach, David N; Waterman, Peter G

    2006-09-01

    The major component (35-65%) of the volatile oil obtained by steam distillation of the leaves of Melaleuca triumphalis has been identified as (rel)-1beta-pentyl-1alpha,6alpha-dihydroxy-3,3,5,5-tetramethylcyclohexa-2,4-dione (trivial name triumphalone). Relative stereochemistry was established by nuclear Overhauser experiments and X-ray studies on the 2-(3,5-dinitrobenzoic acid) derivative. The remainder of the oil was composed of mono- and sesquiterpene hydrocarbons and alcohols. On prolonged standing the presence of a rearrangement product of triumphalone was observed which was characterized as (rel)-1beta-pentyl-1alpha,3alpha-dihydroxy-4,4,6,6-tetramethylcyclohexa-2,5-dione (trivial name isotriumphalone), presumably arising from an acid catalyzed shift of the pentyl group from C-1 to C-2.

  13. Pathogenesis of Goodpasture syndrome: a molecular perspective.

    PubMed

    Borza, Dorin-Bogdan; Neilson, Eric G; Hudson, Billy G

    2003-11-01

    Goodpasture (GP) syndrome is a form of anti-glomerular basement membrane (GBM) disease, in which autoantibodies bind to alpha3(IV) collagen in GBM causing rapidly progressive glomerulonephritis and pulmonary hemorrhage. The conformational GP epitopes have been mapped to 2 regions within the noncollagenous (NC1) domain of the alpha3(IV) chain. Recently, we described the molecular organization of the autoantigen in the native alpha3alpha4alpha5(IV) collagen network of the GBM. The crystal structure of the NC1 domain has revealed how the GP epitopes are sequestered in the native GBM. Further insight into the pathogenesis of disease has been obtained from better animal models. These advances provide a foundation for the development of new specific therapies.

  14. Cell-wall composition and structure of yeast cells and conjugation tubes of Tremella mesenterica.

    PubMed

    Reid, I D; Bartnicki-Garcia, S

    1976-09-01

    Cell walls prepared from vegetative yeast cells and from hormone-induced conjugation tubes of the basidiomycete Tremella mesenterica had similar compositions. Evidence was found for 1,3-alpha-glucan (yeast 38%, tube 25%), 1,3-beta-1,6-beta-glucan (yeast 33%, tube 48%) and chitin (both less than 3%) in the walls. The walls also contained xylose (5 to 7%), mannose (6%), glucuronic acid (approx. 2%), and traces of galactose. Protein amounted to less than 2% of the wall weight. The cell capsule was very insoluble and could not be removed from the cell wall. The conjugation hormone did not appear to exert its effect on cell shape by causing gross changes in wall composition.

  15. High-Pressure Raman and X-ray Diffraction Study of [beta]- and [gamma]-Polymorphs of Aluminum Hydride

    SciTech Connect

    Drozd, Vadym; Garimella, Subrahmanyam; Saxena, Surendra; Chen, Jiuhua; Palasyuk, Taras

    2012-03-26

    Three polymorphs of alane, AlH{sub 3}, ({alpha}, {beta}, and {gamma}) were synthesized and studied at high-pressure in diamond anvil cell by Raman spectroscopy and synchrotron X-ray diffraction. According to synchrotron X-ray diffraction study, {beta}-AlH{sub 3} is stable up to 6 GPa, followed by transformation into {alpha} phase at higher pressures. X-ray-induced decomposition {gamma}-AlH{sub 3} into constituent elements was found at 15 GPa. Raman scattering study at high pressure for both {beta}- and {gamma}-AlH{sub 3} reveals transition into the {alpha} phase with high concentration of structural defects. DFT calculations (VASP code) show that instability of cubic {beta}-alane crystal structure at high pressure is caused by the strong deformation of the [AlH{sub 6}] polyhedra.

  16. KSC-2011-3210

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, a worker inside space shuttle Endeavour's aft section removes an insulating blanket and cover to provide access for the removal and replacement of the Load Control Assembly-2 (LCA-2). Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  17. KSC-2011-3214

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, a worker inside space shuttle Endeavour's aft section removed a cover to provide access for the removal and replacement of the Load Control Assembly-2 (LCA-2). Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  18. KSC-2011-3219

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, technicians inside space shuttle Endeavour's aft section test the Load Control Assembly-2 (LCA-2). Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  19. KSC-2011-3218

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, technicians inside space shuttle Endeavour's aft section test the Load Control Assembly-2 (LCA-2). Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  20. KSC-2011-3220

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Control Center, teams monitor the testing of the Load Control Assembly-2 (LCA-2) located in space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  1. KSC-2011-3216

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A work station, a replacement Load Control Assembly-2 (LCA-2) is being prepared for installation into Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  2. KSC-2011-3212

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, a worker inside space shuttle Endeavour's aft section removes a cover to provide access for the removal and replacement of the Load Control Assembly-2 (LCA-2). Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  3. KSC-2011-3215

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, a worker inside space shuttle Endeavour's aft section is testing the Load Control Assembly-2 (LCA-2) for replacement. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  4. KSC-2011-3260

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, the Load Control Assembly-2 (LCA-2) is removed from inside space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The assembly will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  5. KSC-2011-3217

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A work station, a worker inspects the replacement Load Control Assembly-2 (LCA-2) which is being prepared for installation into Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  6. KSC-2011-3221

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Control Center, teams monitor the testing of the Load Control Assembly-2 (LCA-2) located in space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  7. KSC-2011-3213

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, a worker inside space shuttle Endeavour's aft section removes a cover to provide access for the removal and replacement of the Load Control Assembly-2 (LCA-2). Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  8. KSC-2011-3211

    NASA Image and Video Library

    2011-05-02

    CAPE CANAVERAL, Fla. -- At NASA Kennedy Space Center's Launch Pad 39A, a worker inside space shuttle Endeavour's aft section removes a cover to provide access for the removal and replacement of the Load Control Assembly-2 (LCA-2). Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Kim Shiflett.

  9. KSC-2011-3261

    NASA Image and Video Library

    2011-05-03

    CAPE CANAVERAL, Fla. -- The Load Control Assembly-2 (LCA-2) is outside at NASA Kennedy Space Center's Launch Pad 39A after it was removed from inside space shuttle Endeavour's aft section. Located in the orbiter's aft avionics bay 5, the LCA-2 assembly, which feeds power to the fuel line heaters, is believed to have caused the heaters for Endeavour's auxiliary power unit-1 (APU-1) to fail April 29 during the first launch attempt for the STS-134 mission. The assembly will be replaced and systems will be retested before the launch is rescheduled. STS-134 will deliver the Express Logistics Carrier-3, Alpha Magnetic Spectrometer-2 (AMS), a high-pressure gas tank and additional spare parts for the Dextre robotic helper to the International Space Station. The mission also will be the final spaceflight for Endeavour. For more information, visit www.nasa.gov/mission_pages/shuttle/shuttlemissions/sts134/index.html. Photo credit: NASA/Troy Cryder

  10. [Chemical constituents in the roots of Salvia przewalskii Maxim].

    PubMed

    Chen, Wan-sheng; Jia, Xin-ming; Zhang, Wei-dong; Lou, Zi-yang; Qiao, Chuan-zhuo

    2003-05-01

    To investigate the chemical composition of the root of Salvia przewalskii Maxim. Compounds were isolated by silica gel column chromatography. Structures of these compounds were elucidated by spectral analysis (EI-MS, FAB-MS, 1HNMR, 13CNMR, 1H-1H COSY, 1H-13C COSY, HMBC, NOESY) and phytochemical properties. Eight compounds were isolated and identified as: tanshinone II-A (I), crypotanshinone (II), przewaquinone A (III), sugiol (IV), ursolic acid (V), 2 alpha, 3 alpha-dihydroxy urs-12-ene-28-acid (VI), oleanolic acid (VII), and neo-przewaquinone A (VIII). Compound VIII is a new compound, and compound II, IV, V, VI and VII are isolated from this plant for the first time.

  11. [Separation and preparation of indole alkaloids in Lycorma delicatula White. by HPLC].

    PubMed

    Xue, G; Yuan, S

    1996-09-01

    A HPLC method for separating and preparing indole alkaloids is described. HPLC conditions for analysis: BIO-RAD series 700 HPLC, model 700 data station, UV: model 1749 UV-VIS monitor, column: BIO-RAD Hi-pore RP318, 250 mm x 10 mm, mobile phase: 80% methanol-H2O(gradient), flow rate: 1.5 ml/min, detection wavelength: 254 nm. On the basis of spectral (1HNMR, 13CNMR, H-H COSY, MS, DEPT) and chemical evidence, the structures of two compounds were elucidated as beta-yohimbine (yohimban-16-carboxylic acid-17-hydroxy methylester (3 alpha, 16 alpha, 17 beta)) and ajmalicine (oxayohimban-16-carboxylic acid-16,17-didehydro-19-ethyl methyl ester (19 alpha)).

  12. Sex steroids and personality traits in the middle luteal phase of healthy normally menstruating young professional women.

    PubMed

    Avgoustinaki, Pavlina D; Mitsopoulou, Effrosyni; Chlouverakis, Gregorios; Triantafillou, Theoni; Venihaki, Maria; Koukouli, Sofia; Margioris, Andrew N

    2012-01-01

    Sex steroids affect human behavior. The aim of the present study was to determine the associations, if any, between the circulating levels of gonadal and adrenal sex steroids in the mid luteal phase (21st day of a normal menstrual cycle, MC) of young professional women and psychometric parameters as assessed by the Minnesota Multiphasic Personality Inventory (MMPI). Our results are as follows: (a) The metabolic product of activated adrenal and gonadal androgens, 3alpha-diolG, was modestly but significantly associated with the social introversion scale (10-SI) (r=0.36, p<0.05), independently accounting for 13% of its variation across participants (R²=0.13, F(1,45)=6.58, p=0.014). (b) Total testosterone was significantly associated with the paranoia scale (6-Pa) (r=0.27, p<0.05). Multiple regression analyses indicated that 10% of the variability in paranoia scores could be independently explained by total testosterone levels (R²=0.10, F(1,57)=6.23, p=0.016). We were unable to find any association between the circulating androgens and scores on the masculinity-femininity scale (Mf). We were also unable to document any association between the weak adrenal androgens DHEA and DHEA-S and depression in contrast to several published reports. (c) Our data suggest a marginally significant association between progesterone and scores on the 7-Pt (obsessive/compulsive/psychasthenia) scale (r=0.27, p<0.05). However, only 7% of the 7-Pt variance was explained by progesterone (R²=0.071, F(1,50)=3.81, p=0.057). We have found that total testosterone was associated with the paranoia score, the metabolic product of activated androgens, 3alpha-diolG, to social introversion and, finally, progesterone to obsessive-compulsive behavior.

  13. Steroid modulation of the chloride ionophore in rat brain: structure-activity requirements, regional dependence and mechanism of action

    SciTech Connect

    Gee, K.W.; Bolger, M.B.; Brinton, R.E.; Coirini, H.; McEwen, B.S.

    1988-08-01

    Further in vitro studies of steroids active at the gamma-aminobutyric acidA (GABAA) receptor regulated Cl- channel labeled by (35S)-t-butylbicyclophosphorothionate ((35S)TBPS) reveal additional structural requirements necessary for activity. Evaluation of selected steroids for activity against TBPS-induced convulsions show similar requirements for activity. Interestingly, steroids (e.g., 5 alpha-pregnan-3 alpha, 20 alpha-diol) were identified that have high potency but limited efficacy as modulators of (35S)TBPS binding. These characteristics are reminiscent of the clinically useful benzodiazepines (BZs) such as clonazepam. However, interactions between the prototypical anesthetic-barbiturate, sodium pentobarbital, and steroids active at the Cl- channel suggest that they do not share a common site of action as allosteric modulators of (35S)TBPS and BZ receptor binding. The most potent steroid evaluated, 5 alpha-pregnan-3 alpha-ol-20-one, modulates (35S)TBPS binding at low concentrations (IC50 approximately 17 nM) in a regionally dependent manner. All (35S)TBPS binding sites appear to be functionally coupled to a steroid modulatory site. Because several of the active steroids are metabolites of progesterone, their ability to inhibit the binding of (3H)promegestrone to the cytosolic progestin receptor in rat uterus was evaluated. Those steroids showing potent activity at the GABAA receptor-Cl- ionophore were inactive at the intracellular progestin receptor. Such specificity coupled with their high potency provide additional support for the hypothesis that some of these steroids may be involved in the homeostatic regulation of brain excitability via the GABAA-BZ receptor complex.

  14. HtrA3 as an Early Marker for Preeclampsia: Specific Monoclonal Antibodies and Sensitive High-Throughput Assays for Serum Screening

    PubMed Central

    Dynon, Kemperly; Heng, Sophea; Puryer, Michelle; Li, Ying; Walton, Kelly; Endo, Yaeta; Nie, Guiying

    2012-01-01

    Mammalian HtrA3 (high temperature requirement A3) is a serine protease of the HtrA family. It has two isoforms [long (HtrA3-L) and short (HtrA3-S)] and is important for placental development and cancer progression. Recently, HtrA3 was identified as a potential diagnostic marker for early detection of preeclampsia, a life-threatening pregnancy-specific disorder. Currently there are no high-throughput assays available to detect HtrA3 in human serum. In this study we generated and fully tested a panel of five HtrA3 mouse monoclonal antibodies (mAbs). Three mAbs recognised both HtrA3-L and HtrA3-S and the other two detected HtrA3-L only. All five mAbs were highly specific to HtrA3 and applicable in western blotting and immunohistochemical analysis of endogenous HtrA3 proteins in the mouse and human tissues. Amplified luminescent proximity homogeneous assays-linked immunosorbent assays (AlphaLISAs), were developed to detect HtrA3 isoforms in picomolar levels in serum. The HtrA3 AlphaLISA detected significantly higher serum levels of HtrA3 in women at 13–14 weeks of gestation who subsequently developed preeclampsia compared to gestational-age matched controls. These HtrA3 mAbs are valuable for the development of immunoassays and characterisation of HtrA3 isoform-specific biology. The newly developed HtrA3 AlphaLISA assays are suitable for large scale screening of human serum. PMID:23049902

  15. Characterization of glycoinositolphosphoryl ceramide structure mutant strains of Cryptococcus neoformans.

    PubMed

    Gutierrez, Ana L S; Farage, Layla; Melo, Manuel N; Mohana-Borges, Ronaldo S; Guerardel, Yann; Coddeville, Bernadete; Wieruszeski, Jean-Michel; Mendonça-Previato, Lucia; Previato, Jose O

    2007-06-01

    In fungi, glycoinositolphosphoryl ceramide (GIPC) biosynthetic pathway produces essential molecules for growth, viability, and virulence. In previous studies, we demonstrated that the opportunistic fungus Cryptococcus neoformans synthesizes a complex family of xylose-(Xyl) branched GIPCs, all of which have not been previously reported in fungi. As an effort to understand the biosynthesis of these sphingolipids, we have now characterized the structures of GIPCs from C. neoformans wild-type (KN99alpha) and mutant strains that lack UDP-Xyl, by disruption of either UDP-glucose dehydrogenase (NE321) or UDP-glucuronic acid decarboxylase (NE178). The structures of GIPCs were determined by a combination of nuclear magnetic resonance (NMR) spectroscopy, tandem mass spectrometry (MS), and gas chromatography-MS. The main and largest GIPC from wild-type strain was identified as an alpha-Manp(1 --> 6)alpha-Manp(1 --> 3)alpha-Manp[beta-Xylp(1 --> 2)]alpha-Manp(1 --> 4)beta-Galp(1 --> 6)alpha-Manp(1 --> 2) Ins-1-P-Ceramide, whereas the most abundant GIPC from both mutant strains was found to be an alpha-Manp(1 --> 3)alpha-Manp(1 --> 4)beta-Galp(1 --> 6)alpha-Manp(1 --> 2)Ins-1-P-Ceramide. The ceramide moieties of C. neoformans wild-type and mutant strains were composed of a C(18) phytosphingosine, which was N-acylated with 2-hydroxy tetra-, or hexacosanoic acid, and 2,3-dihydroxy-tetracosanoic acid. Our structural analysis results indicate that the C. neoformans mutant strains are unable to complete the assembly of the GIPC-oligosaccharide moiety due the absence of Xyl side chain.

  16. Sex, drugs and sports: prostaglandins, epitestosterone and sexual development.

    PubMed

    Sanders, Bryan K

    2007-01-01

    Amateau and McCarthy's findings published in Nature Neuroscience (June 2004) are noteworthy for suggesting a role for prostaglandins in sexual development. However, evidence suggests that in manipulating PGE2, they unknowingly implicated 3alpha-hydroxysteroid dehydrogenase [E.C. 1.1.1.50], 3(or 17)alpha-hydroxysteroid dehydrogenase [E.C. 1.1.1.209] and their respective products, androsterone (ADT) and epitestosterone (EpiT), in the developmental masculinization of sex behavior. EpiT is generally regarded as a hormonally inactive 17alpha-epimer of testosterone (T). In rats, the kidney is the primary site of EpiT formation, whereas in humans it originates from the gonads, with only a small contribution secreted by the adrenals. Because the ratio of T to EpiT is nearly constant, it is presently used for assessing steroid abuse in competitive sports, where the World Anti-Doping Agency (WADA) considers a T/EpiT ratio >4 evidence of T doping. Despite its central role in the detection of illict anabolic steroid use, our knowledge of factors effecting EpiT production is poor. Clues in the literature, however, reveal that prostaglandin-mediated processes, such as LHRH release, may influence its production. Antimycotics, NSAIDs, and opioid analgesics used in sports medicine are all known to effect prostaglandin E2 synthesis. Primary PGs are potent inhibitors of ADT oxidation, while indomethacin, a prostaglandin blocker, powerfully inhibits 3alpha-HSD reduction and ADT oxidation. This is significant because ADT inhibits the oxidation of EpiT, and may modulate its antiandrogenic and neuroprotective effects. It is hypothesized that the T/EpiT ratio is increased by COX-2 inhibitors and opiod analgesics, and decreased by antimycotics that do not impair testosterone biosynthesis. Given the devastating personal and career consequences that may result from false positive drug tests, substantive research on the effects of PGE2 manipulations on EpiT is warranted.

  17. Antiviral triterpenoids from the medicinal plant Schefflera heptaphylla.

    PubMed

    Li, Yaolan; Jiang, Renwang; Ooi, Linda S M; But, Paul P H; Ooi, Vincent E C

    2007-05-01

    Schefflera heptaphylla (L.) Frodin is a principal ingredient of an herbal tea formulation widely used for the treatment of common cold in southern China. An extract of the long leafstalk of the compound leaf of S. heptaphylla exhibited the most potent antiviral activity against respiratory syncytial virus (RSV). Further antiviral-guided fractionation and isolation of the leafstalk extract of S. heptaphylla led to obtain two highly active pure triterpenoids, namely 3alpha-hydroxylup-20(29)-ene-23,28-dioic acid and 3-epi-betulinic acid 3-O-sulfate, together with an inactive saponin, 3alpha-hydroxylup-20(29)-ene-23,28-dioic acid 28-O-alpha-l-rhamnopyranosyl-(1-->4)-O-beta-d-glucopyranosyl-(1-->6)-beta-d-glucopyranoside. An antiviral assay using a cytopathic effect (CPE) reduction method showed that the two triterpenoids possessed broader antiviral activity against respiratory syncytial virus (RSV) with a similar 50% inhibition concentration (IC(50)) value of 6.25 microg/mL, influenza A (H1N1) virus with IC(50) values of 25 and 31.3 microg/mL, Coxsackie B3 (Cox B3) virus with IC(50) values of 12.5 and 20 microg/mL and herpes simplex virus type 1 (HSV-1) with IC(50) values of 18.8 and 25 microg/mL, respectively, whereas the saponin did not have antiviral activity against these four viruses at a concentration of 100 microg/mL. Copyright 2007 John Wiley & Sons, Ltd.

  18. Flavonol pentaglycosides of Cordyla (Leguminosae: Papilionoideae: Swartzieae): distribution and taxonomic implications.

    PubMed

    Veitch, Nigel C; Kite, Geoffrey C; Lewis, Gwilym P

    2008-09-01

    A survey of foliar flavonoids in the swartzioid legume genus Cordyla s.l. revealed that three species, C. haraka, C. pinnata and C. richardii, were rich in flavonol pentaglycosides. Their structures were elucidated by spectroscopic and chemical methods as the 3-O-alpha-L-rhamnopyranosyl(1-->3)-alpha-L-rhamnopyranosyl(1-->2)[alpha-L-rhamnopyranosyl(1-->6)]-beta-D-galactopyranoside-7-O-alpha-L-rhamnopyranosides of quercetin and kaempferol (cordylasins A and B, respectively). These compounds were not found in the remaining species, C. africana, C. densiflora, C. madagascariensis (two subspecies) and C. somalensis, which exhibited different profiles of flavonoid glycosides. The distribution of flavonol pentaglycosides in Cordyla s.l. does not support a recent proposal to place both C. haraka and C. madagascariensis in the genus Dupuya [Kirkbride, J.H., 2005. Dupuya, a new genus of Malagasy legumes (Fabaceae). Novon 15, 305-314]. The generic relationship between Cordyla s.l. and Mildbraediodendron is also reassessed on the basis of chemical characters, as the O-linked tetrasaccharide that characterises cordylasins A and B is the same as that found in mildbraedin (kaempferol 3-O-alpha-l-rhamnopyranosyl(1-->3)-alpha-l-rhamnopyranosyl(1-->2)[alpha-l-rhamnopyranosyl(1-->6)]-beta-D-galactopyranoside), the main foliar flavonoid of Mildbraediodendron excelsum. Mildbraedin itself was found to be a minor constituent of leaflet extracts of C. haraka, C. pinnata and C. richardii, and a major constituent of C. somalensis.

  19. Magnetic polyoxometalates: anisotropic exchange interactions in the moiety of [(NaOH2)Co3(H2O)(P2W15O56)2]17-.

    PubMed

    Clemente-Juan, Juan Modesto; Coronado, Eugenio; Gaita-Ariño, Alejandro; Giménez-Saiz, Carlos; Güdel, Hans-Ulrich; Sieber, Andreas; Bircher, Roland; Mutka, Hannu

    2005-05-16

    The magnetic exchange interactions in a C0(3)(11) moiety encapsulated in Na(17) [(NaOH(2))Co(3)(H(2)O)(P(2)W(15)O(56))(2)] (NaCo(3)) were studied by a combination of magnetic measurements (magnetic susceptibility and low-temperature magnetization), with a detailed Inelastic Neutron Scattering (INS) investigation. The novel structure of the salt was determined by X-ray crystallography. The ferromagnetic Co(3)O(14) triangular cluster core consists of three octahedrally oxo-coordinated Co(II) ions sharing edges. According to the single-ion anisotropy and spin-orbit coupling usually assumed for octahedral Co(II) ions, the appropiate exchange Hamiltonian to describe the ground-state properties of the isosceles triangular Co(3) spin cluster is anisotropic and is expressed as H = - 2sigma(alpha)(=)(x,y,z)(J(alpha)(12)S(1alpha)S(2alpha) + J(alpha)(23)S(2alpha)S(3alpha) + J(alpha)(13)S(1alpha)S(3alpha)), where J(alpha) are the components of the exchange interactions between the Co(II) ions. To reproduce the INS data, nonparallel anisotropic exchange tensors needed to be introduced, which were directly connected to the molecular symmetry of the complex. The following range of parameters (value +/- 0.5 cm(-1)) was found to reproduce all experimental information while taking magnetostructural relations into account: J(x)(12) = J(y)(13) = 8.6 cm(-1); J(y)(12) = J(x)(13) = 1.4 cm(-1); J(z)(12) = J(z)(13) = 10.0 cm(-1); J(x)(23) = J(y)(23) = 6.5 cm(-1) and = 3.4 cm(-1).

  20. HIV-1 Efficient Entry in Inner Foreskin Is Mediated by Elevated CCL5/RANTES that Recruits T Cells and Fuels Conjugate Formation with Langerhans Cells

    PubMed Central

    Zhou, Zhicheng; Barry de Longchamps, Nicolas; Schmitt, Alain; Zerbib, Marc; Vacher-Lavenu, Marie-Cécile; Bomsel, Morgane; Ganor, Yonatan

    2011-01-01

    Male circumcision reduces acquisition of HIV-1 by 60%. Hence, the foreskin is an HIV-1 entry portal during sexual transmission. We recently reported that efficient HIV-1 transmission occurs following 1 h of polarized exposure of the inner, but not outer, foreskin to HIV-1-infected cells, but not to cell-free virus. At this early time point, Langerhans cells (LCs) and T-cells within the inner foreskin epidermis are the first cells targeted by the virus. To gain in-depth insight into the molecular mechanisms governing inner foreskin HIV-1 entry, foreskin explants were inoculated with HIV-1-infeceted cells for 4 h. The chemokine/cytokine milieu secreted by the foreskin tissue, and resulting modifications in density and spatial distribution of T-cells and LCs, were then investigated. Our studies show that in the inner foreskin, inoculation with HIV-1-infected cells induces increased CCL5/RANTES (1.63-fold) and decreased CCL20/MIP-3-alpha (0.62-fold) secretion. Elevated CCL5/RANTES mediates recruitment of T-cells from the dermis into the epidermis, which is blocked by a neutralizing CCL5/RANTES Ab. In parallel, HIV-1-infected cells mediate a bi-phasic modification in the spatial distribution of epidermal LCs: attraction to the apical surface at 1 h, followed by migration back towards the basement membrane later on at 4 h, in correlation with reduced CCL20/MIP-3-alpha at this time point. T-cell recruitment fuels the continuous formation of LC-T-cell conjugates, permitting the transfer of HIV-1 captured by LCs. Together, these results reveal that HIV-1 induces a dynamic process of immune cells relocation in the inner foreskin that is associated with specific chemokines secretion, which favors efficient HIV-1 entry at this site. PMID:21738469

  1. Applicability of the dopamine and rate hypotheses in explaining the differences in behavioral pharmacology of the chloro-benztropine analogs: studies conducted using intracerebral microdialysis and population pharmacodynamic modeling.

    PubMed

    Othman, Ahmed A; Newman, Amy Hauck; Eddington, Natalie D

    2007-08-01

    Previous studies indicated that the chloro-benztropine analogs differed significantly in their cocaine-like activity, which was not expected based on the similarity in their in vitro binding affinity and functional potency at the dopamine transporter (DAT). The present study was designed to extend the understanding of the involvement of both pharmacokinetic and pharmacodynamic factors in mediating the behavioral differences among these analogs. The pharmacokinetics of 3'-chloro-3alpha-(diphenylmethoxy)tropane (3'-Cl BZT), the analog showing a cocaine-like behavioral profile in rodents, was compared with previously reported pharmacokinetic characteristics of cocaine and 4',4''-dichloro-3alpha-(diphenylmethoxy)tropane (4',4''-diCl BZT), an analog totally devoid of cocaine-like actions. Microdialysis studies in rats were conducted to determine whether 3'-Cl and 4',4''-diCl BZT differed significantly in their effect on nucleus accumbens extracellular dopamine levels, with cocaine serving as a reference. A mechanistic model based on DAT association/dissociation kinetics was used to describe the time delay between the plasma concentrations of the chloro-analogs and their dopaminergic effects. 3'-Cl BZT had plasma elimination half-life of 1.9 h versus 0.5 and 21.1 h for cocaine and 4',4''-diCl BZT, respectively. 4',4''-diCl BZT increased the DA levels at a slower rate and to a significantly lower extent relative to 3'-Cl BZT that were, in turn, lower than cocaine. The duration of dopamine elevation was as follows: 4',4''-diCl BZT > 3'-Cl BZT > cocaine. The model indicated faster association and dissociation with DAT for 3'-Cl BZT relative to 4',4''-diCl BZT. The present results indicate that behavioral differences among the chloro-analogs may be explainable based on both the dopamine and rate hypotheses of drug abuse.

  2. Formation of ursodeoxycholic acid from chenodeoxycholic acid by a 7 beta-hydroxysteroid dehydrogenase-elaborating Eubacterium aerofaciens strain cocultured with 7 alpha-hydroxysteroid dehydrogenase-elaborating organisms.

    PubMed Central

    MacDonald, I A; Rochon, Y P; Hutchison, D M; Holdeman, L V

    1982-01-01

    A gram-positive, anaerobic, chain-forming, rod-shaped anaerobe (isolate G20-7) was isolated from normal human feces. This organism was identified by cellular morphology as well as fermentative and biochemical data as Eubacterium aerofaciens. When isolate G20-7 was grown in the presence of Bacteroides fragilis or Escherichia coli (or another 7 alpha-hydroxysteroid dehydrogenase producer) and chenodeoxycholic acid, ursodeoxycholic acid produced. Time course curves revealed that 3 alpha-hydroxy-7-keto-5 beta-cholanoic acid produced by B. fragilis or E. coli or introduced into the medium as a pure substance was reduced by G20-7 specifically to ursodeoxycholic acid. The addition of glycine- and taurine-conjugated primary bile acids (chenodeoxycholic and cholic acids) and other bile acids to binary cultures of B. fragilis and G20-7 revealed that (i) both conjugates were hydrolyzed to give free bile acids, (ii) ursocholic acid (3 alpha, 7 beta, 12 alpha-trihydroxy-5 beta-cholanoic acid) was produced when conjugated (or free) cholic acid was the substrate, and (iii) the epimerization reaction was at least partially reversible. Corroborating these observations, an NADP-dependent 7 beta-hydroxysteroid dehydrogenase (reacting specifically with 7 beta-OH-groups) was demonstrated in cell-free preparations of isolate G20-7; production of the enzyme was optimal at between 12 and 18 h of growth. This enzyme, when measured in the oxidative direction, was active with ursodeoxycholic acid, ursocholic acid, and the taurine conjugate of ursodeoxycholic acid (but not with chenodeoxycholic, deoxycholic, or cholic acids) and displayed an optimal pH range of 9.8 to 10.2 Images PMID:6758698

  3. In vivo and in vitro hyperbaric studies in mice suggest novel sites of action for ethanol.

    PubMed

    Davies, D L; Bolger, M B; Brinton, R D; Finn, D A; Alkana, R L

    1999-02-01

    The present study uses increased atmospheric pressure as an ethanol antagonist to test the hypothesis that allosteric coupling pathways in the GABA(A) receptor complex represent initial sites of action for ethanol. This was accomplished using behavioral and in vitro measures to determine the effects of pressure on ethanol and other GABAergic drugs in C57BL/6 and LS mice. Behaviorally, exposure to 12 times normal atmospheric pressure (ATA) of a helium-oxygen gas mixture (heliox) antagonized loss of righting reflex (LORR) induced by the allosteric modulators ethanol and pentobarbital, but did not antagonize LORR induced by the direct GABA agonist 4,5,6,7-tetrahydroisoxazolo-pyridin-3-ol (THIP). Similarly, exposure to 12 ATA heliox antagonized the anticonvulsant effects verses isoniazid of ethanol, diazepam and pentobarbital. Biochemically, exposure to 12 ATA heliox antagonized potentiation of GABA-activated 36Cl-uptake by ethanol, flunitrazepam and pentobarbital in LS mouse brain preparations, but did not alter GABA-activated 36Cl- uptake per se. In contrast to its antagonist effect versus other allosteric modulators, pressure did not antagonize these behavioral or in vitro effects induced by the neuroactive steroid, 3alpha-hydroxy-5beta-pregnan-20-one (3alpha,5beta-P). These findings add to evidence that pressure directly and selectively antagonizes drug effects mediated through allosteric coupling pathways. The results fit predictions, and thus support the hypothesis that allosteric coupling pathways in GABA(A) receptors represent initial sites of action for ethanol. Collectively, the results suggest that there may be common physicochemical and underlying structural characteristics that define ethanol sensitive regions of receptor proteins and/or their associated membranes that can be identified by pressure within (e.g., GABA(A)) and possibly across (e.g., GABA(A), NMDA, 5HT3) receptors.

  4. Inhibition of 3(17)beta-hydroxysteroid dehydrogenase from Pseudomonas testosteroni by steroidal A ring fused pyrazoles.

    PubMed

    Levy, M A; Holt, D A; Brandt, M; Metcalf, B W

    1987-04-21

    Several 2,3- and 3,4-steroidal fused pyrazoles have been investigated as potential inhibitors of NAD(P)H-dependent steroid oxidoreductases. These compounds are proven to be potent, specific inhibitors for 3(17) beta-hydroxysteroid dehydrogenase from Pseudomonas testosteroni with Ki values of 6-100 nM. In contrast, the activities of 3 alpha,20 beta-hydroxysteroid dehydrogenase from Streptomyces hydrogenans, steroid 5 alpha-reductase from rat prostate, and 3 alpha-hydroxysteroid dehydrogenase from rat liver were unaffected by micromolar concentrations of these compounds. Product and dead-end inhibition studies indicate an ordered association to the beta-dehydrogenase with the cofactor binding prior to substrate or inhibitor. From the results of double inhibition experiments, it is proposed that inhibition occurs through formation of an enzyme-NAD+-inhibitor ternate. On the basis of pH profiles of Vm/Km, Vm, and 1/Ki and of absorbance difference spectra, a hypothetical mechanism of inhibition by the steroidal pyrazoles, drawn by analogy from the inhibition of liver alcohol dehydrogenase by alkylpyrazoles [Theorell, H., & Yonetani, T. (1963) Biochem. Z. 338, 537-553; Andersson, P., Kvassman, J. K., Lindström, A., Oldén, B., & Pettersson, G. (1981) Eur. J. Biochem. 113, 549-554], is reconsidered. The pH studies and enzyme modification experiments by diethyl pyrocarbonate suggest the involvement of histidine in binding of the inhibitor. A modified proposal for the structure of the enzyme-NAD+-steroidal pyrazole complex is proposed.(ABSTRACT TRUNCATED AT 250 WORDS)

  5. HIV-1 efficient entry in inner foreskin is mediated by elevated CCL5/RANTES that recruits T cells and fuels conjugate formation with Langerhans cells.

    PubMed

    Zhou, Zhicheng; Barry de Longchamps, Nicolas; Schmitt, Alain; Zerbib, Marc; Vacher-Lavenu, Marie-Cécile; Bomsel, Morgane; Ganor, Yonatan

    2011-06-01

    Male circumcision reduces acquisition of HIV-1 by 60%. Hence, the foreskin is an HIV-1 entry portal during sexual transmission. We recently reported that efficient HIV-1 transmission occurs following 1 h of polarized exposure of the inner, but not outer, foreskin to HIV-1-infected cells, but not to cell-free virus. At this early time point, Langerhans cells (LCs) and T-cells within the inner foreskin epidermis are the first cells targeted by the virus. To gain in-depth insight into the molecular mechanisms governing inner foreskin HIV-1 entry, foreskin explants were inoculated with HIV-1-infeceted cells for 4 h. The chemokine/cytokine milieu secreted by the foreskin tissue, and resulting modifications in density and spatial distribution of T-cells and LCs, were then investigated. Our studies show that in the inner foreskin, inoculation with HIV-1-infected cells induces increased CCL5/RANTES (1.63-fold) and decreased CCL20/MIP-3-alpha (0.62-fold) secretion. Elevated CCL5/RANTES mediates recruitment of T-cells from the dermis into the epidermis, which is blocked by a neutralizing CCL5/RANTES Ab. In parallel, HIV-1-infected cells mediate a bi-phasic modification in the spatial distribution of epidermal LCs: attraction to the apical surface at 1 h, followed by migration back towards the basement membrane later on at 4 h, in correlation with reduced CCL20/MIP-3-alpha at this time point. T-cell recruitment fuels the continuous formation of LC-T-cell conjugates, permitting the transfer of HIV-1 captured by LCs. Together, these results reveal that HIV-1 induces a dynamic process of immune cells relocation in the inner foreskin that is associated with specific chemokines secretion, which favors efficient HIV-1 entry at this site.

  6. Structures of two O-chain polysaccharides of Citrobacter gillenii O9a,9b lipopolysaccharide. A new homopolymer of 4-amino-4,6-dideoxy-D-mannose (perosamine).

    PubMed

    Lipiński, Tomasz; Zatonsky, George V; Kocharova, Nina A; Jaquinod, Michel; Forest, Eric; Shashkov, Alexander S; Gamian, Andrzej; Knirel, Yuriy A

    2002-01-01

    Mild acid degradation of the lipopolysaccharide of Citro- bacter gillenii O9a,9b released a polysaccharide (PS), which was found to consist of a single monosaccharide, 4- acetamido-4,6-dideoxy-d-mannose (d-Rha4NAc, N-acetyl-d-perosamine). PS was studied by methylation analysis and (1)H-NMR and (13)C-NMR spectroscopy, using two-dimensional (1)H,(1)H COSY, TOCSY, NOESY, and H-detected (1)H,(13)C heteronuclear correlation experiments. It was found that PS includes two structurally different polysaccharides: an alpha1-->2-linked homopolymer of N-acetyl-d-perosamine [-->2)-alpha-d-Rhap4NAc-(1-->, PS2] and a polysaccharide composed of tetrasaccharide repeating units (PS1) with the following structure: -->3)-alpha-d-Rhap4NAc-(1-->2)-alpha-d-Rhap4NAc-(1-->2)-alpha-d-Rhap4NAc-(1-->3)-alpha-d-Rhap4 N Ac2Ac-(1--> where the degree of O-acetylation of a 3-substituted Rha4NAc residue at position 2 is approximately 70%. PS could be fractionated into PS1 and PS2 by gel-permeation chromatography on TSK HW-50S. Matrix-assisted laser desorption ionization MS data indicate sequential chain elongation of both PS1 and PS2 by a single sugar unit, with O-acetylation in PS1 beginning at a certain chain length. Anti-(C. gillenii O9a,9b) serum reacted with PS1 in double immunodiffusion and immunoblotting, whereas neither PS2 nor the lipopolysaccharide of Vibrio cholerae O1 with a structurally related O-chain polysaccharide were reactive.

  7. Progestagen metabolites in fetal sheep plasma: the effect of fetal nephrectomy.

    PubMed

    Dolling, M; Seamark, R F

    1979-10-01

    Fetal sheep (100-115 days gestation) were surgically implanted with femoral arterial and venous cannulae and then either sham-operated (control) or bilaterally nephrectomized. Following a 5-day recovery period, fetal blood samples (10 ml/48 h) were taken and the steroid sulphate fraction analysed as trimethylsilyl esters by gas-liquid chromatography (g. l.c.). Three progestagen metabolites were repeatedly detected in plasma samples from control and nephrectomized fetuses and identified by g.l.c.-mass-spectrometric techniques as 5 beta-pregnane-3 beta,20 beta-diol, 5 beta-pregnane-3 beta,20 alpha-diol and 5 beta-pregnane-3 alpha,20 alpha-diol. In three control fetuses the plasma concentration of both 5 beta-pregnane-3 beta,20 beta-diol and -20 alpha-diol showed a steady increase from about 0.5 micrograms/ml at 105 days to about 1.5 and 2-2.5 micrograms/ml, respectively, at 143 days gestation. A study in one fetus indicated that the values then fell precipitously by term (147 days) as plasma cortisol concentrations rose. In contrast, whilst no consistent patterns were seen in their concentration in five nephrectomized fetuses the levels were 2-10 times higher than the control values (0.5-10 micrograms/ml) at all stages. The plasma concentration of 5 beta-pregnane-3 alpha,20 alpha-diol was less perturbed by nephrectomy and only showed a slight increase over control values (0.2-0.5 micrograms/ml). Three sham-operated fetuses which aborted following infection also showed increased plasma concentrations of 5 beta-pregnane-3 beta,20 beta-diol and -20 alpha-diol, similar to the nephrectomized fetuses. It is postulated that high levels of circulating progesterone metabolites may reflect induced increases in adrenal endocrine activity culminating in premature activation of those changes in adrenal function which trigger parturition.

  8. Chronic anabolic-androgenic steroid treatment affects brain GABA(A) receptor-gated chloride ion transport.

    PubMed

    Bitran, D; Hilvers, R J; Frye, C A; Erskine, M S

    1996-01-01

    Previous research in this laboratory has shown that chronic treatment of adult male rats with an anabolic-androgenic steroid (AAS) produced anxiolytic behavior and increased the functional response of cortical gamma-aminobutyric acid(A) (GABA(A)) receptors. The experiments reported here were aimed at further characterizing the effect of chronic AAS exposure on cerebral cortical GABA(A) receptors. Adult male rats were injected with dianabol (1,4-androstadien-17alpha-methyl-17beta-ol-3-one; 10 mg/kg/day, SC) for 4 weeks. A significant decrease in ventral prostate gland weight was found after 2 weeks of dianabol, and returned to control levels 3 and 10 days after steroid discontinuation. Testicular weights decreased throughout the treatment period but reached statistical significance only during the withdrawal period. Serum 3alpha-androstanediol level was marginally increased afer 2 weeks of dianabol injection, and was significantly decreased at 3 and 10 days after withdrawal. GABA-stimulated 36chloride (Cl-) influx in cortical synaptoneurosomes was increased in animals treated with dianabol for 2 and 4 weeks, and remained elevated 3 days after dianabol withdrawal, returning to control levels at withdrawal day 10. The increase in receptor efficacy was associated with a transient increase in receptor sensitivity (inverse of EC50), apparent after 2 weeks of AAS treatment and at withdrawal day 3. In a follow-up experiment, metabolites of dianabol were tested for the in vitro efficacy in potentiating GABA-stimulated Cl- transport. Only 3alpha-androstanedial and androsterone were found to have potent stimulatory effects. The 3beta-reduced metabolites were inactive, as were metabolites that contained a methyl group at the 17alpha position. These results point to significant facilitative effects of dianabol treatment on brain GABA(A) receptors via the metabolic formation of neuroactive steroids.

  9. Glycerol monolaurate prevents mucosal SIV transmission.

    PubMed

    Li, Qingsheng; Estes, Jacob D; Schlievert, Patrick M; Duan, Lijie; Brosnahan, Amanda J; Southern, Peter J; Reilly, Cavan S; Peterson, Marnie L; Schultz-Darken, Nancy; Brunner, Kevin G; Nephew, Karla R; Pambuccian, Stefan; Lifson, Jeffrey D; Carlis, John V; Haase, Ashley T

    2009-04-23

    Although there has been great progress in treating human immunodeficiency virus 1 (HIV-1) infection, preventing transmission has thus far proven an elusive goal. Indeed, recent trials of a candidate vaccine and microbicide have been disappointing, both for want of efficacy and concerns about increased rates of transmission. Nonetheless, studies of vaginal transmission in the simian immunodeficiency virus (SIV)-rhesus macaque (Macacca mulatta) model point to opportunities at the earliest stages of infection in which a vaccine or microbicide might be protective, by limiting the expansion of infected founder populations at the portal of entry. Here we show in this SIV-macaque model, that an outside-in endocervical mucosal signalling system, involving MIP-3alpha (also known as CCL20), plasmacytoid dendritic cells and CCR5(+ )cell-attracting chemokines produced by these cells, in combination with the innate immune and inflammatory responses to infection in both cervix and vagina, recruits CD4(+) T cells to fuel this obligate expansion. We then show that glycerol monolaurate-a widely used antimicrobial compound with inhibitory activity against the production of MIP-3alpha and other proinflammatory cytokines-can inhibit mucosal signalling and the innate and inflammatory response to HIV-1 and SIV in vitro, and in vivo it can protect rhesus macaques from acute infection despite repeated intra-vaginal exposure to high doses of SIV. This new approach, plausibly linked to interfering with innate host responses that recruit the target cells necessary to establish systemic infection, opens a promising new avenue for the development of effective interventions to block HIV-1 mucosal transmission.

  10. Androgen receptor and NFkB expression in human normal and glaucomatous optic nerve head astrocytes in vitro and in experimental glaucoma.

    PubMed

    Agapova, Olga A; Kaufman, Paul L; Hernandez, M Rosario

    2006-06-01

    For several decades, clinical and experimental observations suggested a relationship between steroids and glaucoma; however, the possibility that androgens are also involved in the glaucomatous changes in the optic nerve heads (ONH) has not been explored. Our previous findings that glaucomatous ONH astrocytes synthesize androgen-metabolising enzymes and overproduce a neuroactive androgen, 5alpha-androstane-3alpha, 17beta-diol (3alpha-diol) led us to propose that ONH astrocytes are androgen target cells. Androgens modulate different cellular processes through androgen receptor (AR). NFkB is a transcription factor that positively regulates AR transcription. Here, we analysed AR and NFkB expression in normal and glaucomatous ONH astrocytes in vitro, and in vivo in a monkey model of experimental glaucoma (ExpG) by quantitative real time RT-PCR, Western blotting and immunohistochemistry. We demonstrated that in vitro human glaucomatous ONH astrocytes express AR mRNA and protein at higher levels than normal astrocytes and that in vivo ONH astrocytes from eyes with ExpG showed increased nuclear and cytoplasmic AR immunostaining compared to control eyes. In the retina, retinal ganglion cells (RGC) demonstrated cytoplasmic staining both in control and in ExpG eyes. NFkB mRNA expression was higher in glaucomatous ONH astrocytes than in normal and more nuclear NFkB protein was detected in glaucomatous ONH astrocytes. In vivo immunopositive NFkB nuclear staining of ONH astrocytes in ONH and in RGC in retina was detected both in control and in ExpG eyes. We conclude that in addition to our published data, increase of AR and NFkB expression in glaucomatous ONH astrocytes provides strong evidence that androgens play a significant role in the pathophysiology of glaucoma.

  11. Paraquat exposure reduces nicotinic receptor-evoked dopamine release in monkey striatum.

    PubMed

    O'Leary, Kathryn T; Parameswaran, Neeraja; Johnston, Louisa C; McIntosh, J Michael; Di Monte, Donato A; Quik, Maryka

    2008-10-01

    Paraquat, an herbicide widely used in the agricultural industry, has been associated with lung, liver, and kidney toxicity in humans. In addition, it is linked to an increased risk of Parkinson's disease. For this reason, we had previously investigated the effects of paraquat in mice and showed that it influenced striatal nicotinic receptor (nAChR) expression but not nAChR-mediated dopaminergic function. Because nonhuman primates are evolutionarily closer to humans and may better model the effects of pesticide exposure in man, we examined the effects of paraquat on striatal nAChR function and expression in monkeys. Monkeys were administered saline or paraquat once weekly for 6 weeks, after which nAChR levels and receptor-evoked [(3)H]dopamine ([(3)H]DA) release were measured in the striatum. The functional studies showed that paraquat exposure attenuated dopamine (DA) release evoked by alpha3/alpha6beta2(*) (nAChR that is composed of the alpha3 or alpha6 subunits, and beta2; the asterisk indicates the possible presence of additional subunits) nAChRs, a subtype present only on striatal dopaminergic terminals, with no decline in release mediated by alpha4beta2(*) (nAChR containing alpha4 and beta2 subunits, but not alpha3 or alpha6) nAChRs, present on both DA terminals and striatal neurons. Paraquat treatment decreased alpha4beta2(*) but not alpha3/alpha6beta2(*) nAChR expression. The differential effects of paraquat on nAChR expression and receptor-evoked [(3)H]DA release emphasize the importance of evaluating changes in functional measures. The finding that paraquat treatment has a negative impact on striatal nAChR-mediated dopaminergic activity in monkeys but not mice indicates the need for determining the effects of pesticides in higher species.

  12. Purification and properties of a new testosterone 17beta-dehydrogenase (NADP+) from guinea-pig liver.

    PubMed

    Kaguera, E; Toki, S

    1977-06-01

    As a result of studies of guinea-pig live testosterone 17beta-dehydrogenase (NADP+) (EC 1.1.1.64), a new testosterone 17beta-dehydrogenase was discovered. The new enzyme was purified to a single homogeneous protein from the 105 000 g-supernatant fraction of guinea-pig liver by (NH4)2SO4 fractional precipitation and two gel-filtration stages, DEAE-cellulose column chromatography and hydroxyapatite column chromatography. It was characterized by many properties. The enzyme has almost the same properties as the classical testosterone 17beta-dehydrogenase (NADP+) (EC 1.1.1.64), with respect to cofactor requirement, pH optima for dehydrogenation, effect of phosphate ion on the NAD+-dependent reaction and molecular weight, but characteristic differences were observed in substrate-specificity between the two dehydrogenases. With various androstane derivatives, the configuration of the A/B-ring junction was closely connected with enzyme activity. 5alpha-Androstanes, such as 5alpha-androstane-3alpha,17beta-diol, 5alpha-androstane-3beta,17beta-diol and 17beta-hydroxy-5alpha-androstan-3-one, and 5beta-congeners, such as 5beta-androstane-3alpha,17beta-diol, 5beta-androstane-3beta,17beta-diol and 17beta-hydroxy-5beta-androstan-3-one, served as substrates for both the EC 1.1.1.64 enzyme and the new enzyme. The EC 1.1.1.64 enzyme oxidized testosterone more rapidly than did the new enzyme. These comparisons were based on the relative activities, apparent Km values and apparent Vmax values.

  13. Detection and characterization of a new metabolite of 17alpha-methyltestosterone.

    PubMed

    Pozo, Oscar J; Van Eenoo, Peter; Deventer, Koen; Lootens, Leen; Van Thuyne, Wim; Parr, Maria K; Schänzer, Wilhelm; Sancho, Juan V; Hernández, Felix; Meuleman, Philip; Leroux-Roels, Geert; Delbeke, Frans T

    2009-11-01

    The misuse of the anabolic steroid methyltestosterone is currently routinely monitored in doping control laboratories by gas chromatography-mass spectrometry (GC-MS) of two of its metabolites: 17alpha-methyl-5beta-androstane-3alpha,17beta-diol and 17alpha-methyl-5alpha-androstane-3alpha,17beta-diol. Because of the absence of any easy ionizable moiety, these metabolites are poorly detectable using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization (ESI). In this study, the metabolism of methyltestosterone has been reinvestigated by the use of a precursor ion scan method in LC-ESI-MS/MS. Two metabolites have been detected using this method. Both compounds have been confirmed in postadministration urine samples of an urokinase plasminogen activator-severe combined immunodeficiency (uPA-SCID) mouse with humanized liver and were characterized by LC-MS/MS and GC-MS using both quadrupole and time of flight analyzers. From the detailed study of the fragmentation, these metabolites were proposed to be epimethyltestosterone and a dehydrogenated compound. Epimethyltestosterone has previously been described as a minor metabolite, whereas the occurrence of the oxidized metabolite has not been reported. Comparison with the synthesized reference revealed that the structure of the dehydrogenated metabolite is 6-ene-epimethyltestosterone. A selected reaction monitoring method including three transitions for each metabolite has been developed and applied to samples from an excretion study and to samples declared positive after GC-MS analysis. 6-Ene-epimethyltestosterone was found in all samples, showing its applicability in the detection of methyltestosterone misuse.

  14. Rates of ubiquitin conjugation increase when muscles atrophy, largely through activation of the N-end rule pathway

    NASA Technical Reports Server (NTRS)

    Solomon, V.; Baracos, V.; Sarraf, P.; Goldberg, A. L.

    1998-01-01

    The rapid loss of muscle mass that accompanies many disease states, such as cancer or sepsis, is primarily a result of increased protein breakdown in muscle, and several observations have suggested an activation of the ubiquitin-proteasome system. Accordingly, in extracts of atrophying muscles from tumor-bearing or septic rats, rates of 125I-ubiquitin conjugation to endogenous proteins were found to be higher than in control extracts. On the other hand, in extracts of muscles from hypothyroid rats, where overall proteolysis is reduced below normal, the conjugation of 125I-ubiquitin to soluble proteins decreased by 50%, and treatment with triiodothyronine (T3) restored ubiquitination to control levels. Surprisingly, the N-end rule pathway, which selectively degrades proteins with basic or large hydrophobic N-terminal residues, was found to be responsible for most of these changes in ubiquitin conjugation. Competitive inhibitors of this pathway that specifically block the ubiquitin ligase, E3alpha, suppressed most of the increased ubiquitin conjugation in the muscle extracts from tumor-bearing and septic rats. These inhibitors also suppressed ubiquitination in normal extracts toward levels in hypothyroid extracts, which showed little E3alpha-dependent ubiquitination. Thus, the inhibitors eliminated most of the differences in ubiquitination under these different pathological conditions. Moreover, 125I-lysozyme, a model N-end rule substrate, was ubiquitinated more rapidly in extracts from tumor-bearing and septic rats, and more slowly in those from hypothyroid rats, than in controls. Thus, the rate of ubiquitin conjugation increases in atrophying muscles, and these hormone- and cytokine-dependent responses are in large part due to activation of the N-end rule pathway.

  15. Anatomical Substrate and Scalp EEG Markers are Correlated in Subjects with Cognitive Impairment and Alzheimer's Disease.

    PubMed

    Moretti, Davide V; Frisoni, Giovanni B; Binetti, Giuliano; Zanetti, Orazio

    2011-01-01

    Dementia is a syndromic diagnosis, encompassing various stage of severity and different anatomo-physiological substrates. The hippocampus is one of the first and most affected brain regions affected by both Alzheimer's disease (AD) and mild cognitive impairment (MCI). Moreover, chronic cerebrovascular disease (CVD) is one of the major risk factor for developing dementia. Recent studies have demonstrated different relationship between the anatomical substrate and scalp electroencephalography (EEG) markers. Indeed, modifications of EEG rhythmicity is not proportional to the hippocampal atrophy, whereas changes in EEG activity are directly proportional to the load of subcortical CVD. The computation of the EEG spectral power and the analysis of the functional coupling of brain areas, through linear coherence, are two of the most known processing methods in EEG research. Two specific EEG markers, theta/gamma and alpha3/alpha2 frequency ratio have been reliable associated to the atrophy of amygdalo-hippocampal complex. Moreover, theta/gamma ratio has been related to MCI conversion in dementia and alpha3/alpha2 ratio has been specifically related to MCI conversion in AD. The functional coupling of brain areas is also modulated by hippocampal atrophy. In the MCI subjects, hippocampal atrophy is linked to an increase of interhemispheric coherence seen on frontal and temporal regions whereas subcortical CVD is linked to a decrease of coherence in fronto-parietal regions. In the present study the most significant results of recent studies on correlation between scalp EEG, cognitive decline, and anatomical substrate have been reviewed, with particular attention to the relationships between EEG changes and hippocampal atrophy. The following review is not intended to provide a comprehensive summary of the literature. Rather it identifies and discusses selected studies that are designed to find the specific correlation between scalp EEG markers and anatomo-pathological substrate

  16. The disintegrin domain of ADAM9: a ligand for multiple beta1 renal integrins.

    PubMed

    Mahimkar, Rajeev M; Visaya, Orvin; Pollock, Allan S; Lovett, David H

    2005-01-15

    Renal tubular epithelial cells in all nephron segments express a distinct member of the metalloprotease-disintegrin family, ADAM9 (a disintegrin and metalloprotease 9), in a punctate basolateral distribution co-localized to the beta1 integrin chain [Mahimkar, Baricos, Visaya, Pollock and Lovett (2000) J. Am. Soc. Nephrol. 11, 595-603]. Discrete segments of the nephron express several defined beta1 integrins, suggesting that ADAM9 interacts with multiple renal integrins and thereby regulates epithelial cell-matrix interactions. Intact ADAM9 and a series of deletion constructs sequentially lacking the metalloprotease domain and the disintegrin domain were assembled as chimaeras with a C-terminal GFP (green fluorescent protein) tag. Stable expression of the ADAM9/GFP protein on the surface of HEK-293 cells (human embryonic kidney 293 cells) significantly decreased adhesion to types I and IV collagen, vitronectin and laminin, but had little effect on adhesion to fibronectin. Expression of the disintegrin/cysteine-rich/GFP construct yielded a similar, but more marked pattern of decreased adhesion. Expression of the cysteine-rich/GFP construct had no effect on adhesion, indicating that the disintegrin domain was responsible for the competitive inhibition of cell-matrix binding. To define the specific renal tubular beta1 integrins interacting with the ADAM9 disintegrin domain, a recombinant GST (glutathione S-transferase)-disintegrin protein was used as a substrate in adhesion assays in the presence or absence of specific integrin-blocking antibodies. Inclusion of antibodies to alpha1, alpha3, alpha6, alphav and beta1 blocked adhesion of HEK-293 cells to GST-disintegrin protein. Immobilized GST-disintegrin domain perfused with renal cortical lysates specifically recovered the alpha3, alpha6, alphav and beta1 integrin chains by Western analysis. It is concluded that ADAM9 is a polyvalent ligand, through its disintegrin domain, for multiple renal integrins of the beta1

  17. Determining structure and function of steroid dehydrogenase enzymes by sequence analysis, homology modeling, and rational mutational analysis.

    PubMed

    Duax, William L; Thomas, James; Pletnev, Vladimir; Addlagatta, Anthony; Huether, Robert; Habegger, Lukas; Weeks, Charles M

    2005-12-01

    The short-chain oxidoreductase (SCOR) family of enzymes includes over 6,000 members identified in sequenced genomes. Of these enzymes, approximately 300 have been characterized functionally, and the three-dimensional crystal structures of approximately 40 have been reported. Since some SCOR enzymes are steroid dehydrogenases involved in hypertension, diabetes, breast cancer, and polycystic kidney disease, it is important to characterize the other members of the family for which the biological functions are currently unknown and to determine their three-dimensional structure and mechanism of action. Although the SCOR family appears to have only a single fully conserved residue, it was possible, using bioinformatics methods, to determine characteristic fingerprints composed of 30-40 residues that are conserved at the 70% or greater level in SCOR subgroups. These fingerprints permit reliable prediction of several important structure-function features including cofactor preference, catalytic residues, and substrate specificity. Human type 1 3beta-hydroxysteroid dehydrogenase isomerase (3beta-HSDI) has 30% sequence identity with a human UDP galactose 4-epimerase (UDPGE), a SCOR family enzyme for which an X-ray structure has been reported. Both UDPGE and 3-HSDI appear to trace their origins back to bacterial 3alpha,20beta-HSD. Combining three-dimensional structural information and sequence data on the 3alpha,20beta-HSD, UDPGE, and 3beta-HSDI subfamilies with mutational analysis, we were able to identify the residues critical to the dehydrogenase function of 3-HSDI. We also identified the residues most probably responsible for the isomerase activity of 3beta-HSDI. We test our predictions by specific mutations based on sequence analysis and our structure-based model.

  18. Aromatase inhibitors as add-on treatment for men with epilepsy.

    PubMed

    Harden, Cynthia; MacLusky, Neil J

    2005-01-01

    Manipulation of neurosteroids to treat epilepsy has been an area of active research. The effect of testosterone on brain excitability and seizure threshold has been mixed; the estradiol metabolite of testosterone increases brain excitability, while the reduced metabolite of testosterone, 3alpha-androstanediol, decreases brain excitability, likely through an action at the gamma-amino butyric acid A receptor. Therefore, the metabolites of testosterone produce opposite effects on brain excitability in seizure models. Aromatase is the enzyme for the conversion of testosterone to 17beta-estradiol. Aromatase inhibitors could decrease brain excitability by decreasing local estradiol levels and therefore, could be beneficial for the treatment of epilepsy. Aromatase inhibitors are US Food and Drug Administration-approved and have a long history of safe use in menopausal women with breast cancer. This review presents the results of using anastrazole in an open-label, add-on manner in a small group of men with epilepsy in order to improve seizures. The results suggested some effect on reduction of seizures and no side effects. Testosterone levels did increase, but not to above the normal range. Letrozole used in a single case was also beneficial for seizures. It was concluded that aromatase inhibitors may be a useful adjunct to the treatment of epilepsy, but habituation to the treatment may be limiting. Many men with epilepsy have low testosterone, and aromatase inhibition may be helpful in restoring levels to normal. Modulation of reproductive hormones by aromatase inhibition as well as enhancement of the 3alpha-androstanediol pathway may be an avenue of epilepsy treatment that would not produce sedative side effects, which is often a limiting factor with standard antiseizure medications. A further interesting result is that elevated follicle stimulating hormone and luteal stimulating hormone levels were associated with seizure reduction, suggesting that they may be a

  19. Intra-adipose sex steroid metabolism and body fat distribution in idiopathic human obesity.

    PubMed

    Wake, Deborah J; Strand, Magnus; Rask, Eva; Westerbacka, Jukka; Livingstone, Dawn E W; Soderberg, Stefan; Andrew, Ruth; Yki-Jarvinen, Hannele; Olsson, Tommy; Walker, Brian R

    2007-03-01

    Causes of visceral fat accumulation include glucocorticoid excess or decreased oestrogen/androgen ratio either in plasma or within adipose tissue. In obese subjects, the intra-adipose cortisol-generating enzyme 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) is increased, but information on sex steroid signalling is sparse. We aimed to test associations between body fat or fat distribution and mRNA transcript levels for androgen and oestrogen receptors and for enzymes metabolizing sex steroids in adipose tissue. A cross-sectional study. Forty-five healthy men and women with body mass index (BMI) 21-36 kg/m(2). In subcutaneous adipose biopsies we measured mRNAs for enzymes metabolizing local oestrogens (aromatase) and androgens [5alpha-reductase type 1; AKR1C2 (3alpha-HSD3); AKR1C3 (17beta-HSD5, 3alpha-HSD2)] and for sex steroid receptors [oestrogen receptor (ER)-alpha and androgen receptor (AR)]. We related these to body fat mass and distribution. Generalized obesity (BMI) was associated with increased aromatase mRNA (r = 0.35, P < 0.05). Central obesity (waist : hip ratio) was associated with mRNA for AKR1C2 (r = 0.28, P < 0.05) and AKR1C3 (r = 0.38, P < 0.01) but not aromatase (r = 0.06). 5alpha-Reductase type 1, ER and AR mRNA levels did not predict fat amount or its distribution. These data on transcript levels suggest that, in idiopathic obesity, increased intra-adipose oestrogen generation by aromatase predicts peripheral fat distribution, while androgen metabolism by AKR1C isoforms predicts central fat distribution, supporting the hypothesis that intra-adipose sex steroid metabolism is a determinant of gynoid vs. android patterns of body fat.

  20. Rates of ubiquitin conjugation increase when muscles atrophy, largely through activation of the N-end rule pathway

    NASA Technical Reports Server (NTRS)

    Solomon, V.; Baracos, V.; Sarraf, P.; Goldberg, A. L.

    1998-01-01

    The rapid loss of muscle mass that accompanies many disease states, such as cancer or sepsis, is primarily a result of increased protein breakdown in muscle, and several observations have suggested an activation of the ubiquitin-proteasome system. Accordingly, in extracts of atrophying muscles from tumor-bearing or septic rats, rates of 125I-ubiquitin conjugation to endogenous proteins were found to be higher than in control extracts. On the other hand, in extracts of muscles from hypothyroid rats, where overall proteolysis is reduced below normal, the conjugation of 125I-ubiquitin to soluble proteins decreased by 50%, and treatment with triiodothyronine (T3) restored ubiquitination to control levels. Surprisingly, the N-end rule pathway, which selectively degrades proteins with basic or large hydrophobic N-terminal residues, was found to be responsible for most of these changes in ubiquitin conjugation. Competitive inhibitors of this pathway that specifically block the ubiquitin ligase, E3alpha, suppressed most of the increased ubiquitin conjugation in the muscle extracts from tumor-bearing and septic rats. These inhibitors also suppressed ubiquitination in normal extracts toward levels in hypothyroid extracts, which showed little E3alpha-dependent ubiquitination. Thus, the inhibitors eliminated most of the differences in ubiquitination under these different pathological conditions. Moreover, 125I-lysozyme, a model N-end rule substrate, was ubiquitinated more rapidly in extracts from tumor-bearing and septic rats, and more slowly in those from hypothyroid rats, than in controls. Thus, the rate of ubiquitin conjugation increases in atrophying muscles, and these hormone- and cytokine-dependent responses are in large part due to activation of the N-end rule pathway.

  1. Simplified method to measure glucocorticoid metabolites in faeces of horses.

    PubMed

    Flauger, Birgit; Krueger, Konstanze; Gerhards, Hartmut; Möstl, Erich

    2010-02-01

    Glucocorticoids or their metabolites can be measured in several body fluids or excreta, including plasma, saliva, urine and faeces. In recent years the measurement of glucocorticoid metabolites (GCMs) in faeces has gained increasing attention, because of its suitability for wild populations. In horses, however, the group-specific enzyme immunoassay described so far has a limited practicability due to its complex extraction procedure. Therefore, we tested the applicability of other enzyme immunoassays for glucocorticoid metabolites. The present study clearly proved that an enzyme immunoassay (EIA) for 11-oxoaetiocholanolone using 11-oxoaetiocholanolone-17-CMO: BSA (3alpha,11-oxo-A EIA) as antigen showed high amounts of immunoreactive substances. Therefore it was possible to use just a small amount of the supernatant of a methanolic suspension of faeces. The results correlated well with the already described method for measuring GCMs in horse faeces, i.e. analysing the samples with an EIA after a two step clean up procedure of the samples (Merl et al. 2000). In addition, the 3alpha,11-oxo-A EIA has the advantage of providing a bigger difference between baseline values and peak values after ACTH stimulation. The new assay increased the accuracy of the test, lowered the expenses per sample, and storing samples at room temperature after collection was less critical than with other assays investigated in our study. This is a big advantage both in the field of wildlife management of equids and in the field of equestrian sports and it shows the importance of choosing an assay which is in good accordance with the metabolites excreted in a given species.

  2. Isolation and identification of androstanediol glucuronide from human plasma.

    PubMed

    Rao, P N; Burdett, J E; Moore, P H; Horton, R

    1987-11-01

    [3H]Dihydrotestosterone (50 microCi) was infused into normal men and women for 8 h. It was previously shown that this was sufficient time for this material to reach a steady state. Venous plasma was obtained at 6 and 8 h, pooled, and the unconjugated steroids removed by ether extraction. The remaining plasma was adjusted to pH 4.9 and the steroid conjugate was extracted first with ethyl acetate and then with an ether-ethanol mixture. The extracts were combined and taken to dryness. Steroid sulfates were solvolyzed using dioxane, and the mixture partitioned between ether and 1% NaOH. The aqueous phase was acidified and added to an XAD-2 column, washed with water, and the glucuronide fraction eluted with methanol. The solvent was concentrated and the methanol extract was passed through a C18 Sep-Pak, filtered through an Acrodisc CR and then subjected to gradient high performance liquid chromatography [HPLC] (Nova-Pak C18, KH2PO4, pH 3, and methanol). The fractions containing steroid glucuronides were collected and esterified with diazomethane and then acetylated with acetic anhydride in pyridine. The glucuronide triacetyl methyl ester (GAME) derivatives were then run in a second HPLC system (3 Lichrosorb 5 mu columns, 4 mm x 25 cm) using a gradient of ethanol-heptane and heptane. We clearly established that this system separates 3 alpha-diol GAME conjugated at the 17 and 3 positions (44 vs 50 min) with authentic samples previously synthesized in our laboratory. We concluded that the pooled plasma contained only the 17-GAME conjugate. No significant activity of the 3-glucuronide was detected. The natural compound in circulation, therefore, is 5 alpha-androstane-3 alpha, 17 beta-diol 17-glucuronide.

  3. Triterpene saponins from Chenopodium quinoa Willd.

    PubMed

    Kuljanabhagavad, Tiwatt; Thongphasuk, Piyanut; Chamulitrat, Walee; Wink, Michael

    2008-06-01

    Twenty triterpene saponins (1-20) have been isolated from different parts of Chenopodium quinoa (flowers, fruits, seed coats, and seeds) and their structures have been elucidated by analysis of chemical and spectroscopic data including 1D- and 2D-NMR. Four compounds (1-4) were identified: 3beta-[(O-beta-d-glucopyranosyl-(1-->3)-alpha-l-arabinopyranosyl)oxy]-23-oxo-olean-12-en-28-oic acid beta-d-glucopyranoside (1), 3beta-[(O-beta-d-glucopyranosyl-(1-->3)-alpha-l-arabinopyranosyl)oxy]-27-oxo-olean-12-en-28-oic acid beta-d-glucopyranoside (2), 3-O-alpha-l-arabinopyranosyl serjanic acid 28-O-beta-d-glucopyranosyl ester (3), and 3-O-beta-d-glucuronopyranosyl serjanic acid 28-O-beta-d-glucopyranosyl ester (4). The following known compounds have not previously been reported as saponin constituents from the flowers and the fruits of this plant: two bidesmosides of serjanic acid (5,6), four bidesmosides of oleanolic acid (7-10), five bidesmosides of phytolaccagenic acid (11-15), four bidesmosides of hederagenin (16-19), and one bidesmoside of 3beta,23,30-trihydroxy olean-12-en-28-oic acid (20). The cytotoxicity of these saponins and their aglycones was tested in HeLa cells. Induction of apoptosis in Caco-2 cells by bidesmosidic saponins 1-4 and their aglycones I-III was determined by flow cytometric DNA analysis. The saponins with an aldehyde group were most active. The relationships between structure and cytotoxic activity of saponins and their aglycones are discussed.

  4. Detection of manipulation in doping control urine sample collection: a multidisciplinary approach to determine identical urine samples.

    PubMed

    Thevis, Mario; Geyer, Hans; Mareck, Ute; Sigmund, Gerd; Henke, Jürgen; Henke, Lotte; Schänzer, Wilhelm

    2007-08-01

    Manipulation of urine sampling in sports drug testing is considered a violation of anti-doping rules and is consequently sanctioned by regulatory authorities. In 2003, three identical urine specimens were provided by three different athletes, and the identity of all urine samples was detected and substantiated using numerous analytical strategies including gas chromatography-mass spectrometry with steroid and metabolite profiling, gas chromatography-nitrogen/phosphorus detector analysis, high-performance liquid chromatography-UV fingerprinting, and DNA-STR (short tandem repeat) analysis. None of the respective athletes was the donor of the urine provided for doping analysis, which proved to be a urine sample collected from other unidentified individual(s). Samples were considered suspicious based on identical steroid profiles, one of the most important parameters for specimen individualization in sports drug testing. A database containing 14,224 urinary steroid profiles of athletes was screened for specific values of 4 characteristic parameters (ratios of testosterone/epitestosterone, androsterone/etiocholanolone, androsterone/testosterone, and 5alpha-androstane-3alpha,17beta-diol/5beta-androstane-3alpha,17beta-diol) and only the three suspicious samples matched all criteria. Further metabolite profiling regarding indicated medications and high-performance liquid chromatography-UV fingerprinting substantiated the assumption of manipulation. DNA-STR analyses unequivocally confirmed that the 3 urine samples were from the same individual and not from the athletes who provided DNA from either buccal cell material or blood specimens. This supportive evidence led to punishment of all three athletes according to the rules of the World Anti-Doping Agency. Application of a new multidisciplinary strategy employing common and new doping control assays enables the detection of urine substitution in sports drug testing.

  5. Time-dependent aldosterone metabolism in toad urinary bladder

    SciTech Connect

    Brem, A.S.; Pacholski, M.; Morris, D.J.

    1988-04-01

    Aldosterone (Aldo) metabolism was examined in the toad bladder. Bladders were incubated with (/sup 3/H)aldosterone (10(-7) M) for 5 h, 1 h, or 10 min. Tissues were analyzed for metabolites using high-pressure liquid chromatography (HPLC). In separate experiments, Na+ transport was assessed by the short-circuit current (SCC) technique. Following a 5-h tissue incubation, about 25% of the (/sup 3/H)-aldosterone was converted into metabolites including a polar monosulfate metabolite, 20 beta-dihydroaldo (20 beta-DHAldo), small quantities of 5 beta-reduced products, and a variety of 5 alpha-reduced Aldo products including 5 alpha-DHAldo, 3 alpha,5 alpha-tetrahydroaldo (3 alpha,5 alpha-THAldo), and 3 beta,5 alpha-THAldo. Tissues metabolized approximately 10% of the labeled hormone into the same compounds by 1 h. Measurable quantities of these metabolites were also synthesized by bladders exposed to Aldo for only 10 min and then incubated in buffer for an additional 50 min without Aldo. Bladders pretreated with the spironolactone, K+-canrenoate (3.5 X 10(-4) M), and stimulated with Aldo (10(-7) M) generated a peak SCC 44 +/- 6% of that observed in matched pairs stimulated with Aldo (P less than 0.001; n = 6). K+-canrenoate also markedly diminished (/sup 3/H)aldosterone metabolism at both 5 and 1 h. Thus, metabolic transformation of Aldo begins prior to hormone-induced increases in Na+ transport. Both the generation of certain metabolites (e.g., 5 alpha-reductase pathway products) and the increase in Na+ transport can be selectively inhibited by K+-canrenoate.

  6. Involvement of cytokines in human immunodeficiency virus-1 protein Tat and methamphetamine interactions in the striatum.

    PubMed

    Theodore, Shaji; Cass, Wayne A; Maragos, William F

    2006-06-01

    Human immunodeficiency virus-1 (HIV-1) infection of the brain causes elevation in pro-inflammatory cytokines and inflammatory changes in the striatum. HIV-1-infected individuals who also abuse drugs including the psychostimulant methamphetamine (MA) develop more severe encephalitis and neuronal damage compared to HIV-1-infected patients who do not abuse drugs. In previous studies, we demonstrated that the HIV-1 protein Tat and MA interacted to cause enhanced loss of dopamine in the rat striatum via the destruction of dopaminergic terminals. Since both Tat and MA activate glia and induce cytokine production, we investigated the role of cytokines in the synergistic neurotoxicity induced by Tat and MA using cytokine arrays. Significant increases in monocyte chemotactic protein (MCP-1), interleukin-1 alpha (IL-1alpha) and tissue inhibitor of metalloproteinase-1 (TIMP-1) levels were noted 4 h following Tat + MA treatment compared to saline, Tat or MA. MCP-1 and TIMP-1 levels remained elevated 16 h after Tat + MA compared to saline or MA but were not different from the Tat-treated group at this time point. Weak, but significant elevations in cytokine-induced neutrophil chemoattractant-3 (CINC-3), ciliary neurotrophic factor (CNTF) and macrophage inflammatory protein-3 alpha (MIP-3alpha) were also noted with Tat + MA. The interaction of Tat and MA was prevented in mice genetically deficient in MCP-1 with a consequent attenuation of Tat + MA neurotoxicity. Our findings suggest that HIV-1 infection with concurrent drug abuse might profoundly increase chemokine levels in the striatum resulting in enhanced damage to the dopaminergic system.

  7. Altered expression and activation of signal transducers and activators of transcription (STATs) in hepatitis C virus infection: in vivo and in vitro studies.

    PubMed

    Larrea, E; Aldabe, R; Molano, E; Fernandez-Rodriguez, C M; Ametzazurra, A; Civeira, M P; Prieto, J

    2006-08-01

    Signal transducers and activators of transcription (STATs) play a critical role in antiviral defence. STAT3 is also important in cell protection against inflammatory damage. STAT proteins are activated by interferons and by hepatoprotective cytokines of the interleukin 6 superfamily, including cardiotrophin 1. We analysed the status of STATs in hepatitis C virus (HCV) infected livers and the relationship between expression and activation of STATs and HCV replication in Huh7 cells transfected with HCV genomic replicon. STAT3alpha expression was reduced in HCV infected livers showing an inverse correlation with serum alanine aminotransferase. In patients with HCV infection, nuclear staining for phosphorylated STAT3 was faint in parenchymal cells (although conspicuous in infiltrating leucocytes), in contrast with strong nuclear staining in hepatocytes from control livers. Expression and activation of STAT1 (a factor activated by both interferon (IFN)-alpha and IFN-gamma) were increased in HCV infected livers, particularly in those with high inflammatory activity. Conversely, phosphorylated STAT2 (a factor selectively activated by IFN-alpha) was undetectable in livers with HCV infection, a finding that was associated with marked downregulation of the two functional subunits of the IFN-alpha receptor. HCV replication in Huh7 cells caused STAT3alpha downregulation and blocked STAT3 phosphorylation by either IFN-alpha or cardiotrophin 1. HCV replication in Huh7 cells also inhibited STAT1 and STAT2 activation by IFN-alpha while there was no impairment of STAT1 phosphorylation by the proinflammatory cytokine IFN-gamma. STAT3 is downregulated in HCV infected livers and in Huh7 cells bearing the full length HCV replicon. HCV replication is associated with impaired Jak-STAT signalling by antiviral and cytoprotective cytokines. These effects may favour viral replication while facilitating the progression of liver disease.

  8. Headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for the determination of 5alpha-androst-2-en-17-one and -17beta-ol in the female Asian elephant: application for reproductive monitoring and prediction of parturition.

    PubMed

    Dehnhard, M; Hatt, J-M; Eulenberger, K; Ochs, A; Strauss, G

    2003-02-01

    Asian elephants are not self-sustaining in captivity. The main reasons for this phenomenon are a low birth rate, an aging population, and poor calf-rearing. Therefore, it is essential that reproductive rates had to be improved and there is need for rapid quantitative measures to monitor reproductive functions focussing on estrous detection and the prediction of the period of parturition. The objective of this study was to develop a method which combines headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for analyses of 5alpha-androst-2-en-17beta-ol and -17-one to prognose estrous and to predict the period of parturition. SPME was carried out with a CTC Combi Pal system. The course of the luteal phase-specific substance 5alpha-androst-2-en-17beta-ol and -17-one followed a cyclic pattern in which the follicular and luteal phases could be clearly distinguished (mean estrous cycle length, 15+/-1.4 weeks). Based on daily urine samples, estrous prognosis might be possibly based on the initial 5alpha-androst-2-en-17beta-o1 increase at the end of the follicular phase. Parturition prognosis was performed in three elephant cows based on the 5alpha-androst-2-en-17beta-o1 drop to baseline levels 5-4 days prior parturition. Experiments revealed that 5alpha-androst-3alpha-ol-17-one and probably 5alpha-androst-3alpha-ol-17beta-ol are generated from sulfate conjugates by a thermal process.

  9. Hierarchical phosphorylation at N-terminal transformation-sensitive sites in c-Myc protein is regulated by mitogens and in mitosis.

    PubMed Central

    Lutterbach, B; Hann, S R

    1994-01-01

    The N-terminal domain of the c-Myc protein has been reported to be critical for both the transactivation and biological functions of the c-Myc proteins. Through detailed phosphopeptide mapping analyses, we demonstrate that there is a cluster of four regulated and complex phosphorylation events on the N-terminal domain of Myc proteins, including Thr-58, Ser-62, and Ser-71. An apparent enhancement of Ser-62 phosphorylation occurs on v-Myc proteins having a mutation at Thr-58 which has previously been correlated with increased transforming ability. In contrast, phosphorylation of Thr-58 in cells is dependent on a prior phosphorylation of Ser-62. Hierarchical phosphorylation of c-Myc is also observed in vitro with a specific glycogen synthase kinase 3 alpha, unlike the promiscuous phosphorylation observed with other glycogen synthase kinase 3 alpha and 3 beta preparations. Although both p42 mitogen-activated protein kinase and cdc2 kinase specifically phosphorylate Ser-62 in vitro and cellular phosphorylation of Thr-58/Ser-62 is stimulated by mitogens, other in vivo experiments do not support a role for these kinases in the phosphorylation of Myc proteins. Unexpectedly, both the Thr-58 and Ser-62 phosphorylation events, but not other N-terminal phosphorylation events, can occur in the cytoplasm, suggesting that translocation of the c-Myc proteins to the nucleus is not required for phosphorylation at these sites. In addition, there appears to be an unusual block to the phosphorylation of Ser-62 during mitosis. Finally, although the enhanced transforming properties of Myc proteins correlates with the loss of phosphorylation at Thr-58 and an enhancement of Ser-62 phosphorylation, these phosphorylation events do not alter the ability of c-Myc to transactivate through the CACGTG Myc/Max binding site. Images PMID:8035827

  10. Biovalorization of friedelane triterpenes derived from cork processing industry byproducts.

    PubMed

    Moiteiro, Cristina; Marcelo Curto, Maria João; Mohamed, Nagla; Bailén, María; Martínez-Díaz, Rafael; González-Coloma, Azucena

    2006-05-17

    Here, we describe the synthesis, bioactivity screening, and structure-activity relationships of various synthetic triterpenoids prepared from the cork processing byproducts friedelin (1) and 3-hydroxyfriedel-3-en-2-one (2) via oxidative procedures. The synthesis of compounds 2alpha-trimethylsiloxyfriedelan-3-one (17), friedelin-2,3-lactone (18), friedelin-3-oxime (19), and friedelin-3,4-lactam (20) is also described. We have studied the insecticidal and phytotoxic potential of these compounds, their selective cytotoxic effects on insect and mammalian cells, and their antiparasitic effects. Structural modifications of the A-ring of friedelin (1) improved its insecticidal activity with derivatives 5, 2,3-secofriedelan-2-al-3-oic acid (6), its acetylated derivative 6a, 3beta- and 3alpha-hydroxyfriedelane (9 and 10), 3alpha-hydroxyfriedel-2-one (11), 4beta-hydroxyfriedel-3-one (16), the acetylated 10a, 3,4-secofriedelan-4-oxo-3-oic-acid (14), lactone 18, and the oxime 19 being stronger insecticides than the parent compound. Methyl-3-nor-2,4-secofriedelan-4-oxo-2-oic acid (12) and its acetylated derivative 12a also showed insecticidal activity in contrast to their inactive parent compound 2. The postingestive effects and cytotoxicity of these compounds suggest a multifaceted insecticidal mode of action. These structural modifications did not result in better phytotoxic agents than the parent compounds except for lactam 20 and yielded several moderately active antiparasite derivatives (seco acids 6, 12, 14, and 4beta-hydroxyfriedel-3-one 16) with cytotoxic effects on mammalian cells.

  11. Acceptor products of alternansucrase with gentiobiose. Production of novel oligosaccharides for food and feed and elimination of bitterness.

    PubMed

    Côté, Gregory L

    2009-01-26

    In the presence of suitable acceptor molecules, dextransucrase makes a homologous series of oligosaccharides in which the isomers differ by a single glucosyl unit, whereas alternansucrase synthesizes one trisaccharide, two tetrasaccharides, etc. Previously, we showed that alternansucrase only forms certain isomers of DP>4 from maltose in measurable amounts, and that these oligosaccharides belong to the oligoalternan series rather than the oligodextran series. We now demonstrate that the acceptor products from gentiobiose, also formed in good yields (nearly 90% in unoptimized reactions), follow a pattern similar to those formed from maltose. The initial product is a single trisaccharide, alpha-d-Glcp-(1-->6)-beta-d-Glcp-(1-->6)-d-Glc. Two tetrasaccharides were formed in approximately equal quantities: alpha-d-Glcp-(1-->3)-alpha-d-Glcp-(1-->6)-beta-d-Glcp-(1-->6)-d-Glc and alpha-d-Glcp-(1-->6)-alpha-d-Glcp-(1-->6)-beta-d-Glcp-(1-->6)-d-Glc. Just one pentasaccharide was isolated from the reaction mixture, alpha-d-Glcp-(1-->6)-alpha-d-Glcp-(1-->3)-alpha-d-Glcp-(1-->6)-beta-d-Glcp-(1-->6)-d-Glc. Our hypothesis that the enzyme is incapable of forming two consecutive alpha-(1-->3) linkages, and does not form products with more than two consecutive alpha-(1-->6) linkages, apparently applies to other acceptors as well as to maltose. The glucosylation of gentiobiose reduces or eliminates its bitter taste.

  12. Pyroxene structures, cathodoluminescence and the thermal history of the enstatite chondrites

    NASA Technical Reports Server (NTRS)

    Zhang, Yanhong; Huang, Shaoxiong; Schneider, Diann; Benoit, Paul H.; Sears, Derek W. G.; DeHart, John M.; Lofgren, Gary E.

    1996-01-01

    In order to explore the thermal history of enstatite chondrites, we examined the cathodoluminescence (CL) and thermoluminescence (TL) properties of 15 EH chondrites and 21 EL chondrites, including all available petrographic types, both textural types 3-6 and mineralogical types alpha-delta. The CL properties of EL3(alpha) and EH3(alpha) chondrites are similar. Enstatite grains high in Mn and other transition metals display red CL, while enstatite with low concentrations of these elements show blue CL. A few enstatite grains with greater than 5 wt% FeO display no CL. In contrast, the luminescent properties of the metamorphosed EH chondrites are very different from those of metamorphosed EL chondrites. While the enstatites in metamorphosed EH chondrites display predominantly blue CL, the enstatites in metamorphosed EL chondrites display a distinctive magenta CL with blue and red peaks of approximately equal intensity in their spectra. The TL sensitivities of the enstatite chondrites correlate with the intensity of the blue CL and, unlike other meteorite classes, are not simply related to metamorphism. The different luminescent properties of metamorphosed EH and EL chondrites cannot readily be attributed to compositional differences. But x-ray diffraction data suggests that the enstatite in EH5(gamma),(delta) chondrites is predominantly disordered orthopyroxene, while enstatite in EL6(beta) chondrites is predominantly ordered orthopyroxene. The difference in thermal history of metamorphosed EL and EH chondrites is so marked that the use of single 'petrographic' types is misleading, and separate textural and mineralogical types are preferable. Our data confirm earlier suggestions that metamorphosed EH chondrites underwent relatively rapid cooling, and the metamorphosed EL chondrites cooled more slowly and experienced prolonged heating in the orthopyroxene field.

  13. Glycosidic specificity of fucosyltransferases present in rat epididymal spermatozoa.

    PubMed

    Raychoudhury, S S; Millette, C F

    1995-01-01

    We have recently demonstrated multiple fucosyltransferase (FT) activity in rat spermatogenic cells. To complement these findings, here we identify and partially characterize the glycosidic linkage specificity of FTs present in spermatozoa from caput and cauda epididymides. Analysis of the acceptor substrate specificity of the FTs by thin-layer chromatography indicated that both caput and cauda sperm expressed alpha(1-2)-, alpha(1-3)-, alpha(1-4)-FTs as demonstrated by fucose incorporation into phenyl-beta-D-galactoside, 2'-fucosyllactose, and lacto-N-fucopentaose-I, respectively. Spermatozoa from the cauda epididymidis exhibited significant decreases in the levels of alpha(1-2)-, alpha(1-3)-, alpha(1-4)-FTs, and of total soluble FTs in comparison to spermatozoa from the caput epididymidis. The relative ratio of alpha(1-3)-FT to total FT activity appeared to be significantly higher than those of alpha(1-2)- or alpha(1-4)-FTs, in spermatozoa both from caput and cauda epididymides. Using different types of low molecular weight acceptors and the selective inhibition of the FT by N- ethylmaleimide, we have demonstrated that at least alpha(1-2)-FT is different from alpha(1-3)- or alpha(1-4)-FTs. Kinetic studies also showed that alpha(1-2)-FT is different from alpha(1-3)- or alpha(1-4)-FTs as demonstrated by apparent Km and Vmax values. Moreover, alpha(1-3)- and alpha(1-4)-FT activities in cauda sperm were found to be highly sensitive to Mn2+ but showed differential responses to divalent cations. In contrast, both alpha(1-3)- and alpha(1-4)-FTs seemed to be relatively less sensitive to Mg2+. Thus, these results not only demonstrate the presence of multiple FTs in rat epididymal sperm but also differentiate individual FTs with regard to their kinetic properties and sensitivity to both inhibitor and divalent cations.

  14. Structure of the human gene and two rat cDNAs encoding the alpha chain of GTP-binding regulatory protein Go: two different mRNAs are generated by alternative splicing.

    PubMed Central

    Tsukamoto, T; Toyama, R; Itoh, H; Kozasa, T; Matsuoka, M; Kaziro, Y

    1991-01-01

    Go is a specific class ("other") of signal-transducing heterotrimeric GTP-binding proteins (G proteins) that is expressed in high levels in mammalian brain. We have cloned two different rat cDNAs encoding the alpha subunit of Go (Go alpha-1 and Go alpha-2) and a human Go alpha chromosomal gene. The human Go alpha gene spans more than 100 kilobases and contains 11 exons, including one noncoding exon in the 3' flanking region. The 5' flanking region is highly G + C-rich and contains five G.C boxes (Sp1 binding sites) but no TATA box. Exons 7 and 8 coding for amino acid residues 242-354 of Go alpha protein are duplicated (referred to as exons 7A, 7B, 8A, and 8B). It was found that exons 7A and 8A code for Go alpha-1, and 7B and 8B code for Go alpha-2. This indicates that two different Go alpha mRNAs may be generated by alternative splicing of a single Go alpha gene. The splice sites of the Go alpha-1 and Go alpha-2 genes are completely identical with those encoding human inhibitory G protein alpha subunits Gi2 alpha and Gi3 alpha [Itoh, H., Toyama, R., Kozasa, T., Tsukamoto, T., Matsuoka, M. & Kaziro, Y. (1988) J. Biol. Chem. 263, 6656-6664] and also transducin G protein alpha subunit Gt1 alpha [Raport, C. J., Dere, B. & Hurley, J. (1989) J. Biol. Chem. 264, 7122-7128]. Sequence homology and conservation of the exon-intron organization indicate that the genes coding for Go alpha, Gi2 alpha, Gi3 alpha, Gt1 alpha, and probably Gi1 alpha may be evolved from a common progenitor. Like Go alpha-1, Go alpha-2 is expressed mainly in brain. Images PMID:1901650

  15. a Combined Molecular Dynamics and NMR Spectroscopic Protocol for the Conformational Analysis of Oligosaccharides.

    NASA Astrophysics Data System (ADS)

    Varma, Vikram

    A combined experimental and theoretical protocol for the conformational analysis of oligosaccharides is presented. Three disaccharides, methyl alpha - scD-mannopyranosyl-(1 to 3)-alpha- scD-mannopyranoside, methyl beta- scD-galactopyranosyl-(1 to 4)-beta- scD-glucopyranoside, and propyl beta- scD-2-acetamido -2-deoxy glucopyranosyl-(1 to 3)- alpha- scL-rhamnopyranoside, are used to evaluate a protocol for conformational analysis that makes use of molecular dynamics calculations with the CHARMM force field. Dynamics trajectories computed in vacuo and in water are used to calculate time-averaged NMR parameters such as spin-lattice relaxation times (T_1 ), Nuclear Overhauser Enhancements (NOE), and heteronuclear spin-spin coupling constants (^3J _{rm CH}). The calculated NMR parameters are then compared to experimental values and used to evaluate the computational procedure. The energetically accessible conformations are effectively sampled by the simulations. The method has been extended to the conformational analysis of higher-order oligosaccharides corresponding to the cell-wall polysaccharide of the Streptococcus Group A, and the Shigella flexneri Y O-antigen. The Streptococcus Group A cell-wall polysaccharide is comprised of a backbone of rhamnopyranosyl units connected by alternating alpha- scL-(1 to 3) and alpha- scL -(1 to 2) linkages, to which are attached N-acetyl-beta- scD-glucosamine ( beta- scD-GlcpNAc) residues at the 3 positions of the rhamnose backbone.rm A&rm B^'qquad A^'& rm Bqquad Acr[{-alpha}{-}L{-}Rha {it p}{-}(1to2){-alpha }{-}L{-}Rha{it p} {-}(1to3){-alpha}{ -}L{-}Rha{it p}-(1to2) -alpha-L-Rha{it p}{-}(1 to3){-alpha}{-}L{- }Rha{it p}{-}cr&uparrow(1 to3)&uparrow(1to3)crbeta {-}D{-}&rm Glc{it p }NAcqquadbeta{-}D{-}& rm Glc{it p}NAccr&rm C ^'&rm C] A branched trisaccharide (A^' -(C)B), a tetrasaccharide (A^' -(C)B-A), a pentasaccharide (C^' -B^'-A ^'-(C)B), and two hexasaccharides (C^'-B^ '-A^' -(C)B-A) and (A-(C^')B ^'-A^' -(C)B), have been chosen

  16. Growth hormone regulates the expression of hepatocyte nuclear factor-3 gamma and other liver-enriched transcription factors in the bovine liver.

    PubMed

    Eleswarapu, S; Jiang, H

    2005-01-01

    Growth hormone (GH) regulates the expression of many genes in the liver, and for some genes this regulation may be mediated through liver-enriched transcription factors (LETFs). As part of the long-term goal to investigate the role of LETFs in GH regulation of gene expression in the liver, in this study we determined the effect of GH administration on the expression of 10 LETFs, including hepatocyte nuclear factor (HNF)-1alpha, HNF-1beta, HNF-3alpha, HNF-3beta, HNF-3gamma, HNF-4alpha, HNF-6, CCAAT/enhancer-binding protein (C/EBP) alpha, C/EBPbeta, and albumin D-element binding protein (DBP) in the bovine liver. Eighteen non-lactating and non-pregnant Angus cows were assigned randomly to three groups (n=6 per group) and each cow received a single intramuscular injection of 500 mg slow-release recombinant bovine GH. Liver biopsy samples were taken from group 1 cows 6 h after GH administration, from group 2 cows 24 h after GH administration, and from group 3 cows 1 week after GH administration. Liver biopsies were also collected from group 3 cows 1 day before GH administration, serving as pre-GH controls. The LETF mRNAs in these liver samples were quantified using ribonuclease protection assays with probes generated from bovine LETF cDNAs cloned by standard reverse transcription-polymerase chain reaction. The levels of HNF-3gamma and HNF-6 mRNAs were higher (P< 0.05) in the cows 24 h and 1 week after GH administration than in the untreated cows or the cows 6 h after GH administration. The levels of HNF-4alpha mRNA were higher (P< 0.05) in the cows 1 week after GH administration than in the other three groups of cows. The levels of C/EBPalpha mRNA were higher (P< 0.05) in the cows 24 h after GH administration than in the untreated cows or the cows 6 h after GH administration. The levels of HNF-3alpha mRNA were higher (P< 0.05) in the cows 6 h after GH administration but were lower (P< 0.05) in the cows 24 h or 1 week after GH administration compared with those in the

  17. Detection of dehydroepiandrosterone misuse by means of gas chromatography- combustion-isotope ratio mass spectrometry.

    PubMed

    Mareck, Ute; Geyer, Hans; Flenker, Ulrich; Piper, Thomas; Thevis, Mario; Schänzer, Wilhelm

    2007-01-01

    According to World Anti-Doping Agency (WADA) rules (WADA Technical Document-TD2004EAAS) urine samples containing dehydroepiandrosterone (DHEA) concentrations greater than 100 ng ML(-1) shall be submitted to isotope ratio mass spectrometry (IRMS) analysis. The threshold concentration is based on the equivalent to the glucuronide, and the DHEA concentrations have to be adjusted for a specific gravity value of 1.020. In 2006, 11,012 doping control urine samples from national and international federations were analyzed in the Cologne doping control laboratory, 100 (0.9%) of them yielding concentrations of DHEA greater than 100 ng mL(-1). Sixty-eight percent of the specimens showed specific gravity values higher than 1.020, 52% originated from soccer players, 95% were taken in competition, 85% were male urines, 99% of the IRMS results did not indicate an application of testosterone or related prohormones. Only one urine sample was reported as an adverse analytical finding having 319 ng mL(-1) DHEA (screening result), more than 10,000 ng mL(-1) androsterone and depleted carbon isotope ratio values for the testosterone metabolites androsterone and etiocholanolone. Statistical evaluation showed significantly different DHEA concentrations between specimens taken in- and out-of- competition, whereas females showed smaller DHEA values than males for both types of control. Also a strong influence of the DHEA excretion on different sport disciplines was detectable. The highest DHEA values were detected for game sports (soccer, basketball, handball, ice hockey), followed by boxing and wrestling. In 2007, 6622 doping control urine samples were analyzed for 3alpha,5-cyclo-5alpha-androstan-6beta-ol-17-one (3alpha,5-cyclo), a DHEA metabolite which was described as a useful gas chromatography-mass spectrometry (GC-MS) screening marker for DHEA abuse. Nineteen urine specimens showed concentrations higher than the suggested threshold of 140 ng mL(-1), six urine samples yielded

  18. Identification and characterization of the eps (Exopolysaccharide) gene cluster from Streptococcus thermophilus Sfi6.

    PubMed Central

    Stingele, F; Neeser, J R; Mollet, B

    1996-01-01

    We report the identification and characterization of the eps gene cluster of Streptococcus thermophilus Sfi6 required for exopolysaccharide (EPS) synthesis. This report is the first genetic work concerning EPS production in a food microorganism. The EPS secreted by this strain consists of the following tetrasaccharide repeating unit:-->3)-beta-D-Galp-(1-->3)-[alpha-D-Galp-(1-->6)]-beta-D- D-Galp-(1-->3)-alpha-D-Galp-D-GalpNAc-(1-->. The genetic locus The genetic locus was identified by Tn916 mutagenesis in combination with a plate assay to identify Eps mutants. Sequence analysis of the gene region, which was obtained from subclones of a genomic library of Sfi6, revealed a 15.25-kb region encoding 15 open reading frames. EPS expression in the non-EPS-producing heterologous host, Lactococcus lactis MG1363, showed that within the 15.25-kb region, a region with a size of 14.52 kb encoding the 13 genes epsA to epsM was capable of directing EPS synthesis and secretion in this host. Homology searches of the predicted proteins in the Swiss-Prot database revealed high homology (40 to 68% identity) for epsA, B, C, D, and E and the genes involved in capsule synthesis in Streptococcus pneumoniae and Streptococcus agalactiae. Moderate to low homology (37 to 18% identity) was detected for epsB, D, F, and H and the genes involved in capsule synthesis in Staphylococcus aureus for epsC, D, and E and the genes involved in exopolysaccharide I (EPSI) synthesis in Rhizobium meliloti for epsC to epsJ and the genes involved in lipopolysaccharide synthesis in members of the Enterobacteriaceae, and finally for eps K and lipB of Neisseria meningitidis. Genes (epsJ, epsL, and epsM) for which the predicted proteins showed little or no homology with proteins in the Swiss-Prot database were shown to be involved in EPS synthesis by single-crossover gene disruption experiments. PMID:8626297

  19. Optical studies of nicotinic acetylcholine receptor subtypes in the guinea-pig enteric nervous system.

    PubMed

    Obaid, A L; Nelson, M E; Lindstrom, J; Salzberg, B M

    2005-08-01

    Nicotinic transmission in the enteric nervous system (ENS) is extensive, but the role of individual nicotinic acetylcholine receptor (nAChR) subtypes in the functional connectivity of its plexuses has been elusive. Using monoclonal antibodies (mAbs) against neuronal alpha3-, alpha4-, alpha3/alpha5-, beta2-, beta4- and alpha7-subunits, combined with radioimmunoassays and immunocytochemistry, we demonstrate that guinea-pig enteric ganglia contain all of these nAChR-subunits with the exception of alpha4, and so, differ from mammalian brain. This information alone, however, is insufficient to establish the functional role of the identified nAChR-subtypes within the enteric networks and, ultimately, their specific contributions to gastrointestinal physiology. We have used voltage-sensitive dyes and a high-speed CCD camera, in conjunction with specific antagonists to various nAChRs, to elucidate some of the distinct contributions of the individual subtypes to the behaviour of enteric networks. In the guinea-pig, the submucous plexus has the extraordinary advantage that it is virtually two-dimensional, permitting optical recording, with single cell resolution, of the electrical activity of all of its neurones. In this plexus, the block of alpha3beta2-, alpha3beta4- and/or alpha7-nAChRs always results in a decrease in the magnitude of the synaptic response. However, the magnitude of the fast excitatory post-synaptic potentials (epsps) evoked by electrical stimulation of a neighbouring ganglion varies from cell to cell, reflecting the differential expression of subunits already observed using mAbs, as well as the strengths of the activated synaptic inputs. At the same time, we observe that submucous neurones have a substantial mecamylamine (Mec)-insensitive (non-nicotinic) component to their fast epsps, which may point to the presence of purinergic or serotonergic fast epsps in this system. In the myenteric plexus, on the other hand, the antagonist-induced changes in the

  20. Triterpene glycosides from the stems of Akebia quinata.

    PubMed

    Mimaki, Yoshihiro; Doi, Saya; Kuroda, Minpei; Yokosuka, Akihito

    2007-09-01

    The stems of Akebia quinata have been analyzed for their triterpene glycoside constituents, resulting in the isolation of six new triterpene glycosides, along with 19 known ones. On the basis of extensive spectroscopic analysis, including 2D NMR data, and chemical evidence, the structures of the new compounds were deter-mined to be 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-alpha-L-arabinopyranosyl)oxy]olean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]olean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-alpha-L-arabinopyranosyl)oxy]-23-hydroxyolean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucuronopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]-23-hydroxyolean-12-en-28-oic acid O-alpha-L-rhamnopyranosyl-(1-->4)-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3beta-[(O-beta-D-glucopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]-29-hydroxyolean-12-en-28-oic acid, and 3beta-[(O-beta-D-glucopyranosyl-(1-->3)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl)oxy]-23,29-dihydroxyolean-12-en-28-oic acid, respectively. The main triterpene glycosides contained in the stems of A. quinata were found to have two sugar units at C-3 and C-28 of the aglycone in this study, whereas those of Akebia trifoliate were reported to possess one sugar unit at C-28 of the aglycone. It may be possible to distinguish between A. quinata and A. trifoliate chemically by comparing their triterpene glycoside constituents.

  1. Nicotine induces fibrogenic changes in human liver via nicotinic acetylcholine receptors expressed on hepatic stellate cells

    SciTech Connect

    Soeda, Junpei; Morgan, Maelle; McKee, Chad; Mouralidarane, Angelina; Lin, ChingI; Roskams, Tania; Oben, Jude A.

    2012-01-06

    Highlights: Black-Right-Pointing-Pointer Cigarette smoke may induce liver fibrosis via nicotine receptors. Black-Right-Pointing-Pointer Nicotine induces proliferation of hepatic stellate cells (HSCs). Black-Right-Pointing-Pointer Nicotine activates hepatic fibrogenic pathways. Black-Right-Pointing-Pointer Nicotine receptor antagonists attenuate HSC proliferation. Black-Right-Pointing-Pointer Nicotinic receptor antagonists may have utility as novel anti-fibrotic agents. -- Abstract: Background and aims: Cigarette smoke (CS) may cause liver fibrosis but possible involved mechanisms are unclear. Among the many chemicals in CS is nicotine - which affects cells through nicotinic acetylcholine receptors (nAChR). We studied the effects of nicotine, and involved pathways, on human primary hepatic stellate cells (hHSCs), the principal fibrogenic cells in the liver. We then determined possible disease relevance by assaying nAChR in liver samples from human non-alcoholic steatohepatitis (NASH). Methods: hHSC were isolated from healthy human livers and nAChR expression analyzed - RT-PCR and Western blotting. Nicotine induction of hHSC proliferation, upregulation of collagen1-{alpha}2 and the pro-fibrogenic cytokine transforming growth factor beta 1 (TGF-{beta}1) was determined along with involved intracellular signaling pathways. nAChR mRNA expression was finally analyzed in whole liver biopsies obtained from patients diagnosed with non-alcoholic steatohepatitis (NASH). Results: hHSCs express muscle type ({alpha}1, {beta}1, delta and epsilon) and neuronal type ({alpha}3, {alpha}6, {alpha}7, {beta}2 and {beta}4) nAChR subunits at the mRNA level. Among these subunits, {alpha}3, {alpha}7, {beta}1 and {epsilon} were predominantly expressed as confirmed by Western blotting. Nicotine induced hHSC proliferation was attenuated by mecamylamine (p < 0.05). Additionally, collagen1-{alpha}2 and TGF-{beta}1 mRNA expression were significantly upregulated by nicotine and inhibited by

  2. NMDA potentiation by visible light in the presence of a fluorescent neurosteroid analogue.

    PubMed

    Eisenman, Lawrence N; Shu, Hong-Jin; Wang, Cunde; Aizenman, Elias; Covey, Douglas F; Zorumski, Charles F; Mennerick, Steven

    2009-06-15

    N-Methyl-D-aspartate (NMDA) receptors are widely studied because of their importance in synaptic plasticity and excitotoxic cell death. Here we report a novel method of potentiating NMDA receptors with fluorescence excited by blue (480 nm) light. In the presence of 300 nM of a (7-nitro-2,1,3-benzoxadiazol-4-yl) amino (NBD)-tagged neuroactive steroid carrier C2-NBD-(3alpha,5alpha)-3-hydroxypregnan-20-one (C2-NBD 3alpha5alphaP), responses of cultured hippocampal neurons to 10 microM NMDA were potentiated to 219.2 +/- 9.2% of the baseline response (100%) by a 30 s exposure to 480 nm light. The potentiation decayed back to baseline with a time constant of 80.6 s. Responses to 1 microM and 100 microM NMDA were potentiated to 147.9 +/- 9.6% and 174.1 +/- 15.6% of baseline, respectively, suggesting that visible-light potentiation is relatively insensitive to NMDA concentration. Peak autaptic NMDA responses were potentiated to 178.9 +/- 22.4% of baseline. Similar potentiation was seen with 10 microM NBD-lysine, suggesting that visible-light potentiation is not a steroid effect. Potentiation was also seen with a steroid analogue in which the NBD was replaced with fluorescein, suggesting that NBD is not the only fluorophore capable of supporting visible-light potentiation. UV light and redox potentiation of NMDA receptors largely occluded subsequent blue light potentiation (127.7 +/- 7.4% and 120.2 +/- 6.2% of baseline, respectively). The NR1a(C744A,C798A) mutant that is insensitive to redox and UV potentiation was also largely unaffected by visible-light potentiation (135.0 +/- 10.0% of baseline). Finally, we found that the singlet oxygen scavenger furfuryl alcohol decreased visible-light potentiation. Collectively, these data suggest that visible-light potentiation of NMDA receptors by fluorescence excitation shares mechanisms with UV and redox potentiation and may involve singlet oxygen production.

  3. Separate transport systems for biliary secretion of sulfated and unsulfated bile acids in the rat.

    PubMed Central

    Kuipers, F; Enserink, M; Havinga, R; van der Steen, A B; Hardonk, M J; Fevery, J; Vonk, R J

    1988-01-01

    Biliary secretion of 3 alpha-sulfated bile acids has been studied in Wistar rats with an autosomal recessive defect in the hepatic transport of bilirubin. Liver function, established by measurement of various enzymes in plasma, by enzyme histochemical methods, and by electron microscopy, appeared to be normal in these rats. Serum levels of unconjugated, monoglucuronidated, and diglucuronidated bilirubin were 0.62, 1.62, and 6.16 mumol/liter, respectively, compared with 0.17, 0.08, and 0.02 mumol/liter in control rats. Biliary bilirubin secretion was strongly reduced in the mutant animals: 0.21 +/- 0.03 vs. 0.39 +/- 0.03 nmol/min per 100 g body wt in control rats. Despite normal biliary bile acid output, bile flow was markedly impaired in the mutant animals, due to a 53% reduction of the bile acid-independent fraction of bile flow. The transport maximum for biliary secretion of dibromosulphthalein (DBSP) was also drastically reduced (-53%). Biliary secretion of intravenously administered trace amounts of the 3 alpha-sulfate esters of 14C-labeled taurocholic acid (-14%), taurochenodeoxycholic acid (-39%), taurolithocholic acid (-73%), and glycolithocholic acid (-91%) was impaired in the jaundiced rats compared with controls, in contrast to the biliary secretion of the unsulfated parent compounds. Hepatic uptake of sulfated glycolithocholic acid was not affected in the jaundiced animals. Preadministration of DBSP (15 mumol/100 g body wt) to normal Wistar rats significantly impaired the biliary secretion of sulfated glycolithocholic acid, but did not affect taurocholic acid secretion. We conclude that separate transport systems in the rat liver exist for biliary secretion of sulfated and unsulfated bile acids; the sulfates probably share secretory pathways with the organic anions bilirubin and DBSP. The described genetic defect in hepatic transport function is associated with a reduced capacity to secrete sulfated bile acids into bile; this becomes more pronounced with

  4. Espin cytoskeletal proteins in the sensory cells of rodent taste buds.

    PubMed

    Sekerková, Gabriella; Freeman, David; Mugnaini, Enrico; Bartles, James R

    2005-09-01

    Espins are multifunctional actin-bundling proteins that are highly enriched in the microvilli of certain chemosensory and mechanosensory cells, where they are believed to regulate the integrity and/or dimensions of the parallel-actin-bundle cytoskeletal scaffold. We have determined that, in rats and mice, affinity purified espin antibody intensely labels the lingual and palatal taste buds of the oral cavity and taste buds in the pharyngo-laryngeal region. Intense immunolabeling was observed in the apical, microvillar region of taste buds, while the level of cytoplasmic labeling in taste bud cells was considerably lower. Taste buds contain tightly packed collections of sensory cells (light, or type II plus type III) and supporting cells (dark, or type I), which can be distinguished by microscopic features and cell type-specific markers. On the basis of results obtained using an antigen-retrieval method in conjunction with double immunofluorescence for espin and sensory taste cell-specific markers, we propose that espins are expressed predominantly in the sensory cells of taste buds. In confocal images of rat circumvallate taste buds, we counted 21.5 +/- 0.3 espin-positive cells/taste bud, in agreement with a previous report showing 20.7 +/- 1.3 light cells/taste bud when counted at the ultrastructural level. The espin antibody labeled spindle-shaped cells with round nuclei and showed 100% colocalization with cell-specific markers recognizing all type II [inositol 1,4,5-trisphosphate receptor type III (IP(3)R(3))(,) alpha-gustducin, protein-specific gene product 9.5 (PGP9.5)] and a subpopulation of type III (IP(3)R(3), PGP9.5) taste cells. On average, 72%, 50%, and 32% of the espin-positive taste cells were labeled with antibodies to IP(3)R(3), alpha-gustducin, and PGP9.5, respectively. Upon sectional analysis, the taste buds of rat circumvallate papillae commonly revealed a multi-tiered, espin-positive apical cytoskeletal apparatus. One espin-positive zone, a

  5. Saponins in alfalfa (Medicago sativa L.) root and their structural elucidation.

    PubMed

    Bialy, Z; Jurzysta, M; Oleszek, W; Piacente, S; Pizza, C

    1999-08-01

    Twenty-four saponins have been identified in alfalfa roots, including 13 medicagenic acids, 2 zanhic acids, 4 hederagenins, 1 soyasapogenol A, 2 soyasapogenol B's, 1 soyasapogenol E, and 1 bayogenin glycoside. Ten of the identified compounds, including 3-O-[beta-D-glucopyranosyl(1-->3)-beta-D-glucopyranosyl]-28-O-beta-D- glucopyranoside medicagenate, 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta -D-glucopyranoside] medicagenic acid, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D -glucopyranosyl]-28-beta- D-glucopyranoside medicagenate, 3-O-[beta-D-glucuronopyranosyl methyl ester]-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl(1--> 2)-alpha-L-arabinopyranoside] medicagenate, 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta-D-galactopyranosyl(1-->2)-be ta-D-glucuronopyranosyl]-21-O-alpha-L-rhamnopyranoside soyasapogenol A, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)glucopy ranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyranosyl (1- ->2)-alpha-L-arabinopyranoside] medicagenate, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)glucopy ranosyl]-28-O-¿beta-D-xylopyranosyl(1-->4)-)-[beta-D-apiofurano syl-(1 -->3)]- alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside¿ medicagenate, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D -glucopyranosyl]-28-O-[beta-D-xylopyranosyl(1-->4)-alpha-L-rhamnopyra nosyl(1-->2)-alpha-L-arabinopyranoside] zanhic acid, 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl(1-->2)-beta-D -glucopyranosyl]-28-O-¿beta-D-xylopyranosyl(1-->4)-[beta-D-apiofurano side-(1-->3)]- alpha-L-rhamnopyranosyl(1-->2)-alpha-L-arabinopyranoside¿zanhic acid, and 3-O-[beta-D-galactopyranosyl(1-->2)-beta-D-glucuronopyranosyl]-28- O-b eta-D-glucopyranoside bayogenin, were not reported before, and their structures were established by spectral (FAB-MS and NMR) techniques. In addition, 3-O-[alpha-L-rhamnopyranosyl(1-->2)-beta

  6. Heat-capacity changes in host-guest complexation by Coulomb interactions in aqueous solution.

    PubMed

    Kano, Koji; Ishida, Yoshiyuki; Kitagawa, Kohei; Yasuda, Mayuko; Watanabe, Maki

    2007-10-01

    Heat-capacity changes (deltaC(p)0) were determined for the complexation of 1-alkanecarboxylates with protonated hexakis(6-amino-6-deoxy)-alpha-cyclodextrin (per-NH3(+)-alpha-CD) and heptakis(6-amino-6-deoxy)-beta-cyclodextrin (per-NH3(+)-beta-CD). DeltaC(p)0 decreased with an increase in the binding constant (K) and plateaued at K = 4000 M(-1). The complexes of 1-pentanoate, 1-hexanoate, and 1-heptanoate with per-NH3(+)-alpha-CD are classified as the host-guest system in which the size of the guest fits the CD cavity well. In such a system, van der Waals interaction is the major force for complexation, leading to a negative deltaH0 value. Simultaneously, the water molecules around the hydrophobic alkyl chain of the guest and inside the CD cavity are released to the aqueous bulk phase, leading to a positive deltaS0 value. The negative deltaC(p)0 value in such complexation is ascribed to dehydration of the hydrophobic alkyl chain of the guest and extrusion of the water molecules inside the CD cavity. Meanwhile, the complexes that show positive deltaC(p)0 values are characterized by complexation in which the guest molecules are significantly smaller than the CD cavities. In such a case, the complexation is endothermic and driven by the entropy gain. When the guest is much smaller than the CD cavity, the main binding force should be Coulomb interaction. To form an ionic bond, dehydration of the charged groups must occur. This process is endothermic and leads to positive deltaH0 and deltaS0 values. As the top of the CD cavity is capped by a small but hydrophobic alkyl chain, the water molecules inside the CD cavity may form the iceberg structure. This process must be accompanied by a positive deltaC(p)0 value. Hence, the complexation of a small guest with the CD with a large cavity through Coulomb interactions shows positive and large deltaC(p)0 values. These conclusions were applied to the electrostatic binding of proteins with an anionic ligand.

  7. Binary technetium halides

    NASA Astrophysics Data System (ADS)

    Johnstone, Erik Vaughan

    In this work, the synthetic and coordination chemistry as well as the physico-chemical properties of binary technetium (Tc) chlorides, bromides, and iodides were investigated. Resulting from these studies was the discovery of five new binary Tc halide phases: alpha/beta-TcCl3, alpha/beta-TcCl 2, and TcI3, and the reinvestigation of the chemistries of TcBr3 and TcX4 (X = Cl, Br). Prior to 2009, the chemistry of binary Tc halides was poorly studied and defined by only three compounds, i.e., TcF6, TcF5, and TcCl4. Today, ten phases are known (i.e., TcF6, TcF5, TcCl4, TcBr 4, TcBr3, TcI3, alpha/beta-TcCl3 and alpha/beta-TcCl2) making the binary halide system of Tc comparable to those of its neighboring elements. Technetium binary halides were synthesized using three methods: reactions of the elements in sealed tubes, reactions of flowing HX(g) (X = Cl, Br, and I) with Tc2(O2CCH3)4Cl2, and thermal decompositions of TcX4 (X = Cl, Br) and alpha-TcCl 3 in sealed tubes under vacuum. Binary Tc halides can be found in various dimensionalities such as molecular solids (TcF6), extended chains (TcF5, TcCl4, alpha/beta-TcCl2, TcBr 3, TcI3), infinite layers (beta-TcCl3), and bidimensional networks of clusters (alpha-TcCl3); eight structure-types with varying degrees of metal-metal interactions are now known. The coordination chemistry of Tc binary halides can resemble that of the adjacent elements: molybdenum and ruthenium (beta-TcCl3, TcBr3, TcI 3), rhenium (TcF5, alpha-TcCl3), platinum (TcCl 4, TcBr4), or can be unique (alpha-TcCl2 and beta-TcCl 2) in respect to other known transition metal binary halides. Technetium binary halides display a range of interesting physical properties that are manifested from their electronic and structural configurations. The thermochemistry of binary Tc halides is extensive. These compounds can selectively volatilize, decompose, disproportionate, or convert to other phases. Ultimately, binary Tc halides may find application in the nuclear fuel

  8. Expression of neuronal nicotinic acetylcholine receptor subunit mRNAs in rat hippocampal GABAergic interneurons.

    PubMed

    Son, Jong-Hyun; Winzer-Serhan, Ursula H

    2008-11-10

    Hippocampal inhibitory interneurons are a diverse population of cells widely scattered in the hippocampus, where they regulate hippocampal circuit activity. The hippocampus receives cholinergic projections from the basal forebrain, and functional studies have suggested the presence of different subtypes of nicotinic acetylcholine receptors (AChRs) on gamma-aminobutyric acid (GABA)ergic interneurons. Single-cell polymerase chain reaction analysis had confirmed that several nAChR subunit mRNAs are co-expressed with glutamate decarboxylase 67 (GAD67), the marker for GABAergic interneurons. In this anatomical study, we systematically investigated the co-expression of GAD67 with different nAChR subunits by using double in situ hybridization with a digoxigenin-labeled GAD67 probe and (35)S-labeled probes for nAChR subunits (alpha2, alpha3, alpha4, alpha5, alpha6, alpha7, beta2, beta3, and beta4). The results revealed that most GAD67-positive interneurons expressed beta2, and 67 % also expressed alpha7 mRNA. In contrast, mRNA expression of other subunits was limited; only 13 % of GAD67-positive neurons co-expressed alpha4, and less than 10% expressed transcripts for alpha2, alpha3, alpha5, or beta4. Most GAD67/alpha2 co-expression was located in CA1/CA3 stratum oriens, and GAD67/alpha5 co-expression was predominantly detected in CA1/CA3 stratum radiatum/lacunosum moleculare and the dentate gyrus. Expression of alpha6 and beta3 mRNAs was rarely detected in the hippocampus, and mRNAs were not co-expressed with GAD67. These findings suggest that the majority of nicotinic responses in GABAergic interneurons should be mediated by a homomeric alpha7 or heteromeric alpha7*-containing nAChRs. Other possible combinations such as alpha2beta2*, alpha4beta2*, or alpha5beta2* heteromeric nAChRs could contribute to functional nicotinic response in subsets of GABAergic interneurons but overall would have a minor role.

  9. [Influence of the ovarian cycle on the central nervous system].

    PubMed

    Kuhl, H

    2002-04-01

    Estradiol, progesterone and some of their metabolites modulate the activity of neurotransmitters and neuropeptides in the CNS. The distribution and concentrations of sex steroids in the various CNS regions is partly dependent on the serum levels, but also on the local synthesis of the steroids. In general, estradiol and testosterone exert a stimulatory, progesterone an inhibitory effect on neuronal activities which are mediated by excitatory (e.g. glutamate, aspartate), and inhibitory amino acids (e.g. GABA) and neuropeptides (e.g. beta-endorphin), respectively. Gonadotropin release is primarily governed by the rhythm of pulsatile secretion of GnRH in the hypothalamus which is controlled by estradiol and progesterone by means of inhibitory or stimulatory modulation of the amplitude and frequency of GnRH pulses. The discharges of GnRH neurons triggered by excitatory amino acids are modulated by estradiol, while the inhibitory effect of progesterone is mediated by GABA and beta-endorphin which cause hyperpolarization of the GnRH neurons and consequently a reduced pulse frequency. The pulse amplitudes are primarily influenced by estradiol, but neuropeptide Y, neurotensin and noradrenaline contribute to their preovulatory enhancement. The postovulatory rise in core temperature is caused by the increasing level of progesterone and its metabolite 3 alpha-pregnanolone, respectively. Despite of this, up to 20% of ovulatory cycles do not show any rise in body temperature. Although 3 alpha-pregnanolone has sedative activities, there is no change in sleep quality during the luteal phase due to their low serum levels. It could be demonstrated that performance on tests of articulatory and fine motor skills are enhanced in the late follicular phase as compared to the menstruation phase, while spatial ability was better during menses. Estrogens may influence mood and well-being in a favorable manner, while in predisposed women progesterone may cause symptoms of premenstrual

  10. Millisecond autothermal catalytic reforming of carbohydrates for synthetic fuels by reactive flash volatilization

    NASA Astrophysics Data System (ADS)

    Dauenhauer, Paul Jakob

    Carbohydrates including glucose, cellulose, starch and polyols including glycerol, ethylene glycol and methanol produced in large quantities from biomass are considered as a carbon-based feedstock for high temperature catalytic reforming by catalytic partial oxidation. Autothermal catalytic partial oxidation of methanol, ethylene glycol, and glycerol with Rh and Pt-based catalysts with ceria on alumina foam supports at residence times less than ten milliseconds produced equilibrium selectivity to synthesis gas. The addition of steam at S/C>4 produced selectivity to H2 higher than 80% with little or no selectivity to minor products. In a new process referred to as 'reactive flash volatilization,' catalytic partial oxidation was combined with pyrolysis of biomass by directly impinging particles of cellulose, starch, polyethylene, soy oil, or Aspen (Populous Tremuloides) on an operating Rh-based reforming catalyst at 700-800°C. Solid particles endothermically pyrolyzed to volatile organic compounds which mixed with air and reformed on the catalyst exothermically generating heat to drive the overall process. Particles of ˜250 mum microcrystalline cellulose processed at the conditions of C/O=1.0 on a RhCe/gamma-Al2O3/alpha-Al 2O3 at a residence time of ˜70 milliseconds produced a gaseous effluent stream selecting for 50% H2 and 50% CO with no observable side products other than H2O and CO2, and <1% CH4. To obtain a more optimal synthesis gas stream, the reforming of ˜400 mum microcrystalline particles was examined over a fixed bed of RhCe/gamma-Al2O3/alpha-Al2O 3 spheres by varying the feed ratio of N2/O2, the temperature of the feed gas, the total particle feed rate, and the addition of steam permitting cellulose conversion with ˜75% fuel efficiency. Cellulose, sucrose, and glycerol particle conversion was examined with high-speed photography (1000 frames/second) revealing the formation of a liquid intermediate from cellulose permitting extremely high heat flux (

  11. A phospho-BAD BH3 helix activates glucokinase by a mechanism distinct from that of allosteric activators

    PubMed Central

    Szlyk, Benjamin; Braun, Craig R.; Ljubicic, Sanda; Patton, Elaura; Bird, Gregory H.; Osundiji, Mayowa A.; Matschinsky, Franz M.; Walensky, Loren D.; Danial, Nika N.

    2014-01-01

    Glucokinase is a glucose-phosphorylating enzyme that regulates insulin release and hepatic metabolism, and its loss-of-function is implicated in the pathogenesis of diabetes. Glucokinase activators (GKAs) are attractive therapeutics in diabetes, however, clinical data indicate that their benefits can be offset by hypoglycemia due to marked allosteric enhancement of the enzyme’s glucose affinity. We show that a phospho-mimetic of the BCL-2 homology 3 (BH3) alpha-helix derived from human BAD, a glucokinase binding partner, increases the enzyme catalytic rate without dramatically changing glucose affinity, providing a new mechanism for pharmacologic activation of glucokinase. Remarkably, BAD BH3 phospho-mimetic mediates these effects by engaging a novel region near the enzyme’s active site. This interaction increases insulin secretion in human islets and restores the function of naturally-occurring human glucokinase mutants at the active site. Thus, BAD phospho-mimetics may serve as a novel class of GKAs. PMID:24317490

  12. Hypersecretion of luteinizing hormone and ovarian steroids in women with recurrent early miscarriage.

    PubMed

    Watson, H; Kiddy, D S; Hamilton-Fairley, D; Scanlon, M J; Barnard, C; Collins, W P; Bonney, R C; Franks, S

    1993-06-01

    Polycystic ovary syndrome is associated with hypersecretion of luteinizing hormone (LH) which has been implicated in the aetiology of early pregnancy loss. Although 82% of women with recurrent early loss have polycystic ovaries on ultrasound imaging, random serum LH concentrations are normal. In the present study, we have obtained further information from serial samples concerning the cyclical patterns of gonadotrophin and sex steroid secretion in these women. Twenty-one women with recurrent early pregnancy loss and 10 multiparous controls were investigated; 81% of them and one of ten control subjects had polycystic ovaries. Mean mid-follicular and mid-luteal serum LH and follicle stimulating hormone (FSH) levels were similar in both groups. Seventeen women with pregnancy loss had either raised urinary LH excretion or a premature LH surge; one control subject had a premature LH surge. Total LH excretion during the cycle and mean follicular phase serum testosterone was significantly greater with early pregnancy loss than in the control group, the difference in LH being greatest in the early luteal phase. Urinary oestrone-3-glucuronide excretion was raised in the early luteal phase of the cycle in the group with early pregnancy loss; there was no difference between the groups in pregnanediol-3 alpha-glucuronide excretion. These data demonstrate abnormalities in LH secretion in 81% of women with recurrent fetal loss. Inappropriately raised LH levels may have adverse effects on the developing oocyte or endometrium either directly, or indirectly by causing an elevation in testosterone and oestrogen levels.

  13. Differential tumor microenvironment in human ovarian cystic tumors.

    PubMed

    Tavares Murta, Beatriz Martins; Cunha, Fernando de Queiróz; Miranda, Rodrigo; Adad, Sheila Jorge; Murta, Eddie Fernando Candido

    2004-01-01

    Cells and soluble mediators obtained from tumor effusions are useful in evaluating the tumor microenvironment. Our aim was to examine cytologically and to quantify the leukocyte infiltrate, nitric oxide, cytokines and tumor markers in the intracystic fluid from patients with a cystic adnexal mass, for a possible differentiation between benign and malignant findings. Sixty-six women who had their cystic fluids collected were prospectively divided into benign tumor (22, 33.3%), malignant tumor (10, 15.2%) or other gynecological alterations (34, 51.5%). Cytology, total and differential leukocyte counts were determined by light microscopy. Tumor markers, cytokines and nitric oxide were assayed in the supernatants using the Immulite system, ELISA and Griess reaction, respectively. The sensitivity and specificity of the cytological analysis was 66.7% and 97.7%, respectively. The levels of CA 19.9, CA 15.3, alpha-fetoprotein, carcinoembryonic antigen, progesterone and beta-HCG were significantly higher in the benign and/or malignant group than in the other gynecological alterations. Also, the local concentrations of CA 15.3 and beta-HCG were significantly higher in malignant than in benign tumors. In malignant tumors, increased leukocyte counts and higher concentrations of IL-6, IL-10 and nitric oxide were detected than in benign tumors or other gynecological alterations. In malignant tumors, the microenvironment could be differentiated from benign tumors or other gynecological alterations by cystic fluid analysis.

  14. Theoretical and UV spectral study of isomeric 1-(quinolinyl)-beta-carbolines conformations.

    PubMed

    Eshimbetov, A G; Tulyaganov, T S

    2007-07-01

    On the basis of beta-carboline (1) and 1-(quinolin-2'-yl)-beta-carboline (3) alpha- and t-band energies differences (Delta(alpha,t)) a equilibrium conformations of 1-(quinolin-4'(5'-8')-yl)-beta-carbolines (4-8) in solution have been estimated. Furthermore, as an example of model compounds 1-(alpha'-naphtyl)-beta-carboline (MC1) and 1-(beta'-naphtyl)-beta-carboline (MC2) and also 5 and 6 by molecular mechanics (mm+), semi-empirical (AM1) and none empirical (RHF/6-31G(d)) methods a computations of internal rotation of quinoline fragment around single bond have been performed. It was found that the greater bathochromic shift of the long-wavelength band maxima of 1, in the case of 3 (Deltalambda(max)=39 nm) relatively to 6, 7 (Deltalambda(max)=17+/-2 nm) and 4, 5, 8 (Deltalambda(max)=9+/-1 nm) caused by coplanarity of the molecule 3. Also, from experimental and theoretical investigations a less dihedral angle between beta-carboline and quinoline nucleous for 6, and 7 than 4, 5 and 8 owing to steric and electronic interactions have been found.

  15. Integrin alpha chains exhibit distinct temporal and spatial localization patterns in epithelial cells of the Drosophila ovary.

    PubMed

    Dinkins, Michael B; Fratto, Victoria M; Lemosy, Ellen K

    2008-12-01

    Integrins are heterodimeric transmembrane receptors that modulate cell adhesion, migration, and signaling. Multiple integrin chains contribute to development and morphogenesis of a given tissue. Here, we analyze the expression of Drosophila integrin alpha chains in the ovarian follicular epithelium, a model for tissue morphogenesis and cell migration. We find expression throughout development of the beta chain, betaPS. Alpha chains, however, exhibit both spatial and temporal expression differences. alphaPS1 and alphaPS2 integrins are detected during early and mid-oogenesis on apical, lateral, and basal membranes with the betaPS chain, whereas alphaPS3-family integrins (alphaPS3, alphaPS4, alphaPS5) are expressed in anterior cells late in oogenesis. Surprisingly, we find that alphaPS3-family integrins are dispensable for dorsal appendage morphogenesis but play a role in the final length of the egg, suggesting redundant functions of integrins in a simple tissue. We also demonstrate roles for alphaPS3betaPS integrin in border cell migration and in stretch cells.

  16. Evaluating the suitability of nicotinic acetylcholine receptor antibodies for standard immunodetection procedures.

    PubMed

    Moser, N; Mechawar, N; Jones, I; Gochberg-Sarver, A; Orr-Urtreger, A; Plomann, M; Salas, R; Molles, B; Marubio, L; Roth, U; Maskos, U; Winzer-Serhan, U; Bourgeois, J-P; Le Sourd, A-M; De Biasi, M; Schröder, H; Lindstrom, J; Maelicke, A; Changeux, J-P; Wevers, A

    2007-07-01

    Nicotinic acetylcholine receptors play important roles in numerous cognitive processes as well as in several debilitating central nervous system (CNS) disorders. In order to fully elucidate the diverse roles of nicotinic acetylcholine receptors in CNS function and dysfunction, a detailed knowledge of their cellular and subcellular localizations is essential. To date, methods to precisely localize nicotinic acetylcholine receptors in the CNS have predominantly relied on the use of anti-receptor subunit antibodies. Although data obtained by immunohistology and immunoblotting are generally in accordance with ligand binding studies, some discrepancies remain, in particular with electrophysiological findings. In this context, nicotinic acetylcholine receptor subunit-deficient mice should be ideal tools for testing the specificity of subunit-directed antibodies. Here, we used standard protocols for immunohistochemistry and western blotting to examine the antibodies raised against the alpha3-, alpha4-, alpha7-, beta2-, and beta4-nicotinic acetylcholine receptor subunits on brain tissues of the respective knock-out mice. Unexpectedly, for each of the antibodies tested, immunoreactivity was the same in wild-type and knock-out mice. These data imply that, under commonly used conditions, these antibodies are not suited for immunolocalization. Thus, particular caution should be exerted with regards to the experimental approach used to visualize nicotinic acetylcholine receptors in the brain.

  17. Valproic acid, a molecular lead to multiple regulatory pathways.

    PubMed

    Kostrouchová, M; Kostrouch, Z; Kostrouchová, M

    2007-01-01

    Valproic acid (2-propyl pentanoic acid) is a drug used for the treatment of epilepsy and bipolar disorder. Although very rare, side effects such as spina bifida and other defects of neural tube closure indicate that valproic acid interferes with developmental regulatory pathways. Recently obtained data show that valproic acid affects cell growth, differentiation, apoptosis and immunogenicity of cultured cancer cells and tumours. Focused studies uncovered the potential of valproic acid to interfere with multiple regulatory mechanisms including histone deacetylases, GSK3 alpha and beta, Akt, the ERK pathway, the phosphoinositol pathway, the tricarboxylic acid cycle, GABA, and the OXPHOS system. Valproic acid is emerging as a potential anticancer drug and may also serve as a molecular lead that can help design drugs with more specific and more potent effects on the one side and drugs with wide additive but weaker effects on the other. Valproic acid is thus a powerful molecular tool for better understanding and therapeutic targeting of pathways that regulate the behaviour of cancer cells.

  18. Mechanisms of stem cell osteogenic differentiation on TiO2 nanotubes.

    PubMed

    Yu, Weiqiang; Qian, Chao; Jiang, Xinquan; Zhang, Fuqiang; Weng, Weimin

    2015-12-01

    TiO2 nanotubes could stimulate osteogenic differentiation of stem cells, but the molecular mechanisms underlying the interactions between nanotubes and stem cells remain unclear. In this study, we investigated the response of bone marrow stromal cells to nanotubes of different diameters using microarray-based bioinformatics approach. Gene ontology (GO) and GO enrichment network analysis indicated that larger TiO2 nanotubes were more potent than smaller nanotubes in inducing the expression of genes involved in cell proliferation, differentiation, and immune responses, and inhibiting that of genes responsible for cell adhesion. The analysis of the signaling network containing significantly affected genes suggested that Na(+)/K(+) transporting ATPases ATP1A2 (alpha 2 polypeptide) and ATP1A3 (alpha 3 polypeptide), and MAP3K11 (mitogen-activated protein kinase kinase kinase 11) were important for inducing osteogenic differentiation of bone marrow stromal cells without additional osteogenic stimuli. The upregulation of the ATP1A2 and MAP3K11 genes confirmed by real-time PCR indicates that the response of bone marrow stromal cells to nanotube cues may be mediated by the pathways previously implicated in transducing mechanical stress signals. Our results revealed some molecular mechanisms by which TiO2 nanotubes may direct osteogenic differentiation of stem cells. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Overview of the magnetic properties experiments on the Mars Exploration Rovers

    USGS Publications Warehouse

    Madsen, M.B.; Goetz, W.; Bertelsen, P.; Binau, C.S.; Folkmann, F.; Gunnlaugsson, H.P.; Hjollum, J.I.; Hviid, S.F.; Jensen, J.; Kinch, K.M.; Leer, K.; Madsen, D.E.; Merrison, J.; Olsen, M.; Arneson, H.M.; Bell, J.F.; Gellert, Ralf; Herkenhoff, K. E.; Johnson, J. R.; Johnson, M.J.; Klingelhofer, G.; McCartney, E.; Ming, D. W.; Morris, R.V.; Proton, J.B.; Rodionov, D.; Sims, M.; Squyres, S. W.; Wdowiak, T.; Yen, A. S.

    2009-01-01

    The Mars Exploration Rovers have accumulated airborne dust on different types of permanent magnets. Images of these magnets document the dynamics of dust capture and removal over time. The strongly magnetic subset of airborne dust appears dark brown to black in Panoramic Camera (Pancam) images, while the weakly magnetic one is bright red. Images returned by the Microscopic Imager reveal the formation of magnetic chains diagnostic of magnetite-rich grains with substantial magnetization (>8 Am2 kg-1). On the basis of M??ssbauer spectra the dust contains magnetite, olivine, pyroxene, and nanophase oxides in varying proportions, depending on wind regime and landing site. The dust contains a larger amount of ferric iron (Fe3+/Fe tot ??? 0.6) than rocks in the Gusev plains (???0.1-0.2) or average Gusev soil (???0.3). Alpha Particle X-Ray Spectrometer data of the dust show that some of the iron in magnetite is substituted by titanium and chromium. The good correlation of the amount of calcium and sulfur in the dust may be caused by the presence of a calcium sulfate related phase. The overall mineralogical composition points to a basaltic origin of the airborne dust, although some alteration has taken place as indicated by the large degree of oxidation. Copyright 2009 by the American Geophysical Union.

  20. Wet hydrogen peroxide catalytic oxidation of phenol with FeAC (iron-embedded activated carbon) catalysts.

    PubMed

    Liou, Rey-May; Chen, Shih-Hsiung; Huang, Cheng-Hsien; Hung, Mu-Ya; Chang, Jing-Song; Lai, Cheng-Lee

    2010-01-01

    This investigation aims at exploring the catalytic oxidation activity of iron-embedded activated carbon (FeAC) and the application for the degradation of phenol in the wet hydrogen peroxide catalytic oxidation (WHPCO). FeAC catalysts were prepared by pre-impregnating iron in coconut shell with various iron loadings in the range of 27.5 to 46.5% before they were activated. The FeAC catalysts were characterised by measuring their surface area, pore distribution, functional groups on the surface, and X-ray diffraction patterns. The effects of iron loading strongly inhibited the pore development of the catalyst but benefited the oxidation activity in WHPCO. It was found that the complete conversion of phenol was observed with all FeAC catalysts in oxidation. High level of chemical oxygen demand (COD) abatement can be achieved within the first 30 minutes of oxidation. The iron embedded in the activated carbon showed good performance in the degradation and mineralisation of phenol during the oxidation due to the active sites as iron oxides formed on the surface of the activated carbon. It was found that the embedding irons were presented in gamma-Fe(2)O(3), alpha-Fe(2)O(3), and alpha-FeCOOH forms on the activated carbon. The aging tests on FeAC catalysts showed less activity loss, and less iron leaching was found after four oxidation runs.

  1. Role of Triton X-100 in chemiluminescent enzyme immunoassays capable of diagnosing genetic disorders.

    PubMed

    Chong, Richard; Rho, Jee-Eun R; Yoon, Hye-Joo; Park, Paul S; Rho, Tae-Ho D; Park, Jee Y; Park, Lucienne; Kim, Young-Hwan; Lee, Ji Hoon

    2013-11-15

    The use of Triton X surfactants in developing 1,1'-oxalylimidazole chemiluminescent enzyme immunoassays (ODI CEIs) with extended linear response range for the quantification of unconjugated estriol (uE3), alpha-fetoprotein (AFP), and human chorionic gonadotropin (hCG) is reported for the first time. The wider linear dynamic range in ODI CLEIA results from Triton X series (e.g., Triton X-100, -114, -405, -705) acting as an inhibitor in the interaction between Amplex Red (hydrophobic substrate) and horseradish peroxidase (hydrophilic enzyme) to produce resorufin (hydrophobic fluorescent dye). Triton X-100 acts as the appropriate inhibitor in ODI CLEIA. The maximum concentrations of AFP and hCG quantified with sandwich ODI CLEIA in the presence of Triton X-100 were 8 times higher than when analyzed with the same system in the absence of Triton X-100. In addition, the lowest concentration of uE3 determined using competitive ODI CLEIA in the presence of Triton X-100 was 20 times lower than that measured with competitive ODI CLEIA in the absence of Triton X-100. These results indicate that rapid quantification of AFP, uE3, and hCG using cost effective and highly sensitive ODI CLEIAs in the presence of Triton X-100 can be applied as an accurate, precise, and reproducible method to diagnose genetic disorders (e.g., trisomy 18 and trisomy 21) in fetuses. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. In vitro metabolism of radioactive progesterone and testosterone by the gonads of the protandrous Rhabdosargus sarba at various sexual phases

    SciTech Connect

    Yeung, W.S.; Chan, S.T.

    1985-08-01

    The in vitro steroidogenic capacity of the gonadal tissue in the protandrous Rhabdosargus sarba was studied. Testicular and ovarian tissues from various sexual phases were used either separately or combined. With progesterone as precursor, high yield of 5 beta-reduced metabolites, and no 11-ketotestosterone or 11 beta-hydroxytestosterone were found. The production of 5 beta-pregnan-3 alpha-ol-20-one and 5 beta-pregnane-3,20-dione was very high in incubations with testicular tissue from intersexes or males and was low in those with ovarian tissue only. The production of 17 alpha-hydroxyprogesterone was high in the female but was low in other sexual phases. With testosterone as precursor, 11-ketotestoterone and 11 beta-hydroxytestosterone were identified. 5 beta-Reductase activity was high in the male and relatively low in the female. 5 alpha-Reduced products, however, were absent. There was an increase in the production of androstenedione as the animal underwent sex reversal. The significance of this change in steroidogenesis in this protandrous fish is at present under investigation. Experimental results also indicated that in the intersexual gonad there may be interaction between the testicular component and the ovarian component in steroidgenesis.

  3. Overexpression of TEAD-1 in transgenic mouse striated muscles produces a slower skeletal muscle contractile phenotype.

    PubMed

    Tsika, Richard W; Schramm, Christine; Simmer, Gretchen; Fitzsimons, Daniel P; Moss, Richard L; Ji, Juan

    2008-12-26

    TEA domain (TEAD) transcription factors serve important functional roles during embryonic development and in striated muscle gene expression. Our previous work has implicated a role for TEAD-1 in the fast-to-slow fiber-type transition in response to mechanical overload. To investigate whether TEAD-1 is a modulator of slow muscle gene expression in vivo, we developed transgenic mice expressing hemagglutinin (HA)-tagged TEAD-1 under the control of the muscle creatine kinase promoter. We show that striated muscle-restricted HA-TEAD-1 expression induced a transition toward a slow muscle contractile protein phenotype, slower shortening velocity (Vmax), and longer contraction and relaxation times in adult fast twitch extensor digitalis longus muscle. Notably, HA-TEAD-1 overexpression resulted in an unexpected activation of GSK-3alpha/beta and decreased nuclear beta-catenin and NFATc1/c3 protein. These effects could be reversed in vivo by mechanical overload, which decreased muscle creatine kinase-driven TEAD-1 transgene expression, and in cultured satellite cells by TEAD-1-specific small interfering RNA. These novel in vivo data support a role for TEAD-1 in modulating slow muscle gene expression.

  4. Alcohol absorption inhibitors from bay leaf (Laurus nobilis): structure-requirements of sesquiterpenes for the activity.

    PubMed

    Yoshikawa, M; Shimoda, H; Uemura, T; Morikawa, T; Kawahara, Y; Matsuda, H

    2000-08-01

    Through a bioassay-guided separation using inhibitory activity on blood ethanol elevation in oral ethanol-loaded rat, various sesquiterpenes having an alpha-methylene-gamma-butyrolactone moiety, costunolide (1), dehydrocostus lactone (2), zaluzanin D (3), reynosin (4), santamarine (5), 3alpha-acetoxyeudesma-1,4(15),11(13)-trien-12,6alpha-+ ++olide (6) and 3-oxoeudesma-1,4,11(13)-trien-12,6alpha-olide (7), were isolated as the active principle from the leaves of Laurus nobilis (bay leaf, laurel). In order to characterize the structure requirement for the activity, several reduction products (2a-2d) and amino acid adducts (2e, 2f) of the alpha-methylene-gamma-butyrolactone moiety were synthesized from 2 and the inhibitory activities of these sesquiterpenes, together with alpha-methylene-gamma-butyrolactone (12) and its related compounds (13-16), were examined. These results indicated that the gamma-butyrolactone or gamma-butyrolactol moiety having alpha-methylene or alpha-methyl group was essential for the inhibitory activity on ethanol absorption. Since 1, 2 and 12 showed no significant effect on glucose absorption, these sesquiterpenes appeared to selectively inhibit ethanol absorption. In addition, the acute toxicities of 1 and 2 in a single oral administration were found to be lower than that of 12.

  5. Darwin's evolution theory, brain oscillations, and complex brain function in a new "Cartesian view".

    PubMed

    Başar, Erol; Güntekin, Bahar

    2009-01-01

    Comparatively analyses of electrophysiological correlates across species during evolution, alpha activity during brain maturation, and alpha activity in complex cognitive processes are presented to illustrate a new multidimensional "Cartesian System" brain function. The main features are: (1) The growth of the alpha activity during evolution, increase of alpha during cognitive processes, and decrease of the alpha entropy during evolution provide an indicator for evolution of brain cognitive performance. (2) Human children younger than 3 years are unable to produce higher cognitive processes and do not show alpha activity till the age of 3 years. The mature brain can perform higher cognitive processes and demonstrates regular alpha activity. (3) Alpha activity also is significantly associated with highly complex cognitive processes, such as the recognition of facial expressions. The neural activity reflected by these brain oscillations can be considered as constituent "building blocks" for a great number of functions. An overarching statement on the alpha function is presented by extended analyzes with multiple dimensions that constitute a "Cartesian Hyperspace" as the basis for oscillatory function. Theoretical implications are considered.

  6. Balancing omega-6 and omega-3 fatty acids in ready-to-use therapeutic foods (RUTF).

    PubMed

    Brenna, J Thomas; Akomo, Peter; Bahwere, Paluku; Berkley, James A; Calder, Philip C; Jones, Kelsey D; Liu, Lei; Manary, Mark; Trehan, Indi; Briend, André

    2015-05-15

    Ready-to-use therapeutic foods (RUTFs) are a key component of a life-saving treatment for young children who present with uncomplicated severe acute malnutrition in resource limited settings. Increasing recognition of the role of balanced dietary omega-6 and omega-3 polyunsaturated fatty acids (PUFA) in neurocognitive and immune development led two independent groups to evaluate RUTFs. Jones et al. (BMC Med 13:93, 2015), in a study in BMC Medicine, and Hsieh et al. (J Pediatr Gastroenterol Nutr 2015), in a study in the Journal of Pediatric Gastroenterology and Nutrition, reformulated RUTFs with altered PUFA content and looked at the effects on circulating omega-3 docosahexaenoic acid (DHA) status as a measure of overall omega-3 status. Supplemental oral administration of omega-3 DHA or reduction of RUTF omega-6 linoleic acid using high oleic peanuts improved DHA status, whereas increasing omega-3 alpha-linolenic acid in RUTF did not. The results of these two small studies are consistent with well-established effects in animal studies and highlight the need for basic and operational research to improve fat composition in support of omega-3-specific development in young children as RUTF use expands.

  7. Neurosteroid metabolism in the human brain.

    PubMed

    Stoffel-Wagner, B

    2001-12-01

    This review summarizes the current knowledge of the biosynthesis of neurosteroids in the human brain, the enzymes mediating these reactions, their localization and the putative effects of neurosteroids. Molecular biological and biochemical studies have now firmly established the presence of the steroidogenic enzymes cytochrome P450 cholesterol side-chain cleavage (P450SCC), aromatase, 5alpha-reductase, 3alpha-hydroxysteroid dehydrogenase and 17beta-hydroxysteroid dehydrogenase in human brain. The functions attributed to specific neurosteroids include modulation of gamma-aminobutyric acid A (GABAA), N-methyl-d-aspartate (NMDA), nicotinic, muscarinic, serotonin (5-HT3), kainate, glycine and sigma receptors, neuroprotection and induction of neurite outgrowth, dendritic spines and synaptogenesis. The first clinical investigations in humans produced evidence for an involvement of neuroactive steroids in conditions such as fatigue during pregnancy, premenstrual syndrome, post partum depression, catamenial epilepsy, depressive disorders and dementia disorders. Better knowledge of the biochemical pathways of neurosteroidogenesis and their actions on the brain seems to open new perspectives in the understanding of the physiology of the human brain as well as in the pharmacological treatment of its disturbances.

  8. Regulation of naïve fetal T-cell migration by the chemokines Exodus-2 and Exodus-3.

    PubMed

    Christopherson, K; Brahmi, Z; Hromas, R

    1999-08-03

    We and other workers have recently isolated three novel CC chemokines termed Exodus-1/LARC/Mip-3alpha, Exodus-2/6Ckine/SLC/TCA4, and Exodus-3/Mip-3beta/CKbeta11/ELC. These chemokines share an amino terminal Asp-Cys-Cys-Leu sequence, unique among all chemokines. They also selectively regulate migration of adult T cells. Indeed, there is evidence that Exodus-2 and -3 are critical for adult T-cell adhesion to high endothelial venules in lymph nodes, a rate-limiting step for T-cell trafficking through nodal tissue. Less is known of the factors controlling migration of naïve human fetal T cells. We tested whether these chemokines could regulate chemotaxis in cord blood T-cell populations, and compared that efficacy with normal peripheral blood adult T cells. The findings indicated that naive CD45RA+ cord blood T-cell migration is stimulated by Exodus-2 and -3, and CD4+ cord blood T cells are attracted preferentially by Exodus-2 or -3 as compared with CD8+. Exodus-2 and -3 are likely to be critical in regulating the flux of naive CD4 + fetal T-cell population of secondary lymphoid tissue.

  9. What do we really know about the ubiquitin-proteasome pathway in muscle atrophy?

    NASA Technical Reports Server (NTRS)

    Jagoe, R. T.; Goldberg, A. L.

    2001-01-01

    Studies of many different rodent models of muscle wasting have indicated that accelerated proteolysis via the ubiquitin-proteasome pathway is the principal cause of muscle atrophy induced by fasting, cancer cachexia, metabolic acidosis, denervation, disuse, diabetes, sepsis, burns, hyperthyroidism and excess glucocorticoids. However, our understanding about how muscle proteins are degraded, and how the ubiquitin-proteasome pathway is activated in muscle under these conditions, is still very limited. The identities of the important ubiquitin-protein ligases in skeletal muscle, and the ways in which they recognize substrates are still largely unknown. Recent in-vitro studies have suggested that one set of ubquitination enzymes, E2(14K) and E3(alpha), which are responsible for the 'N-end rule' system of ubiquitination, plays an important role in muscle, especially in catabolic states. However, their functional significance in degrading different muscle proteins is still unclear. This review focuses on the many gaps in our understanding of the functioning of the ubiquitin-proteasome pathway in muscle atrophy, and highlights the strengths and limitations of the different experimental approaches used in such studies.

  10. Predicted unfolding order of the 13 alpha-helices in the catalytic domain of glucoamylase from Aspergillus awamori var. X100 by molecular dynamics simulations.

    PubMed

    Liu, Hsuan-Liang; Wang, Wen-Chi

    2003-01-01

    The unfolding mechanism of the 13 alpha-helices in the catalytic domain of Aspergillus awamori var. X100 glucoamylase was investigated by 200 ps molecular dynamics simulations in explicit water with temperature jump technique. Rather than a simultaneous event, the unfolding of these 13 alpha-helices followed a random ordered mechanism as alpha8-->alpha1-->alpha11-->alpha7-->alpha10-->alpha3-->alpha12-->alpha13-->alpha4-->alpha5-->alpha9-->alpha6-->alpha2. No significant relationships were found between the unfolding order and the length and the hydrophobicity of the helix. alpha-Helix 8 located in the inner region of the catalytic domain was predicted to be the first helix to unfold, indicating that the destruction of the secondary structure motif was initiated from the inner region of the catalytic domain. The dynamic behavior of these alpha-helices induced by increased kinetic energy during the unfolding process is considered to be similar to the expansion and compression of a series of springs under the influence of mechanical stress.

  11. Human periodontal ligament cells facilitate leukocyte recruitment and are influenced in their immunomodulatory function by Th17 cytokine release.

    PubMed

    Konermann, A; Beyer, M; Deschner, J; Allam, J P; Novak, N; Winter, J; Jepsen, S; Jäger, A

    2012-01-01

    The objective of this in vitro study was to examine the immunomodulatory impact of human periodontal ligament (PDL) cells on the nature and magnitude of the leukocyte infiltrate in periodontal inflammation, particularly with regard to Th17 cells. PDL cells were challenged with pro-inflammatory cytokines (IL-1ß, IL-17A, and IFN-γ) and analyzed for the expression of cytokines involved in periodontal immunoinflammatory processes (IL-6, MIP-3 alpha, IL-23A, TGFß1, IDO, and CD274). In order to further investigate a direct involvement of PDL cells in leukocyte function, co-culture experiments were conducted. The expression of the immunomodulatory cytokines studied was significantly increased under pro-inflammatory conditions in PDL cells. Although PDL cells did not stimulate leukocyte proliferation or Th17 differentiation, these cells induced the recruitment of leukocytes. The results of our study suggest that PDL cells might be involved in chronic inflammatory mechanisms in periodontal tissues and thus in the transition to an adaptive immune response in periodontitis.

  12. A serendipitous discovery of antifreeze protein-specific activity in C-linked antifreeze glycoprotein analogs.

    PubMed

    Eniade, Adewale; Purushotham, Madhusudhan; Ben, Robert N; Wang, J B; Horwath, Kathleen

    2003-01-01

    Structurally diverse carbon-linked (C-linked) analogs of antifreeze glycoprotein (AFGP) have been prepared via linear or convergent solid phase synthesis. These analogs range in molecular weight from approx 1.5-4.1 KDa and do not possess the beta-D-galactose-1,3-alpha-D-N-acetylgalactosamine carbohydrate moiety or the L-threonine-L-alanine-L-alanine polypeptide backbone native to the AFGP wild-type. Despite these dramatic structural modifications, the 2.7-KDa and 4.1-KDa analogs possess antifreeze protein-specific activity as determined by recrystallization-inhibition (RI) and thermal hysteresis (TH) assays. These analogs are weaker than the wild-type in their activity, but nanoliter osmometry indicates that these compounds are binding to ice and affecting a localized freezing point depression. This is the first example of a C-linked AFGP analog that possesses TH and RI activity and suggests that the rational design and synthesis of chemically and biologically stable AFGP analogs is a feasible and worthwhile endeavor. Given the low degree of TH activity, these compounds may prove useful for the protection of cells during freezing and thawing cycles.

  13. Extended protection capabilities of an immature dendritic-cell targeting malaria sporozoite vaccine.

    PubMed

    Luo, Kun; Zavala, Fidel; Gordy, James; Zhang, Hong; Markham, Richard B

    2017-03-22

    Mouse studies evaluating candidate malaria vaccines have typically examined protective efficacy over the relatively short time frames of several weeks after the final of multiple immunizations. The current study examines the protective ability in a mouse model system of a novel protein vaccine construct in which the adjuvant polyinosinic polycytidilic acid (poly(I:C)) is used in combination with a vaccine in which the immature dendritic cell targeting chemokine, macrophage inflammatory protein 3 alpha (MIP3α), is fused to the circumsporozoite protein (CSP) of Plasmodium falciparum (P. falciparum). Two vaccinations, three weeks apart, elicited extraordinarily high, MIP3α-dependent antibody responses. MIP3α was able to target the vaccine to the CCR6 receptor found predominantly on immature dendritic cells and significantly enhanced the cellular influx at the vaccination site. At three and 23 weeks after the final of two immunizations, mice were challenged by intravenous injection of 5×10(3) transgenic Plasmodium berghei sporozoites expressing P. falciparum CSP, a challenge dose approximately one order of magnitude greater than that which is encountered after mosquito bite in the clinical setting. A ninety-seven percent reduction in liver sporozoite load was observed at both time points, 23 weeks being the last time point tested.

  14. The nature and origin of spontaneous noise in G protein-gated ion channels

    PubMed Central

    1991-01-01

    Arrival of agonist is generally thought to initiate the signal transduction process in G protein-receptor coupled systems. However, the muscarinic atrial K+ (K+[ACh]) channel opens spontaneously in the absence of applied agonist, giving a noisy appearance to the current records. We investigated the nature and origin of the noise by measuring single channel currents in cell-attached or excised, inside- out membrane patches. Guanosine triphosphate (GTP) produced identical single channel currents in a concentration- and Mg(2+)-dependent manner in the presence or absence of carbachol, but the requirements for GTP were greater in the absence of agonist. Hence the agonist-independent currents appeared to be produced by an endogenous G protein, Gk. This prediction was confirmed when an affinity-purified, sequence-specific Gi-3 alpha antibody or pertussis toxin (PTX) blocked the agonist- independent currents. Candidate endogenous agonists were ruled out by the lack of effect of their corresponding antagonists. Thus agonist- independent currents had the same nature as agonist-dependent K+[ACh] currents and seemed to originate in the same way. We have developed a hypothesis in which agonist-free, empty receptors prime Gk with GTP and Gk activates atrial K+ [ACh] channels producing basal currents or noise. Agonist-independent activation by G proteins of effectors including ion channels appears to be a common occurrence. PMID:1651979

  15. CaV3.2 is the major molecular substrate for redox regulation of T-type Ca2+ channels in the rat and mouse thalamus.

    PubMed

    Joksovic, Pavle M; Nelson, Michael T; Jevtovic-Todorovic, Vesna; Patel, Manoj K; Perez-Reyes, Edward; Campbell, Kevin P; Chen, Chien-Chang; Todorovic, Slobodan M

    2006-07-15

    Although T-type Ca(2+) channels in the thalamus play a crucial role in determining neuronal excitability and are involved in sensory processing and pathophysiology of epilepsy, little is known about the molecular mechanisms involved in their regulation. Here, we report that reducing agents, including endogenous sulfur-containing amino acid l-cysteine, selectively enhance native T-type currents in reticular thalamic (nRT) neurons and recombinant Ca(V)3.2 (alpha1H) currents, but not native and recombinant Ca(V)3.1 (alpha1G)- and Ca(V)3.3 (alpha1I)-based currents. Consistent with this data, T-type currents of nRT neurons from transgenic mice lacking Ca(V)3.2 channel expression were not modulated by reducing agents. In contrast, oxidizing agents inhibited all native and recombinant T-type currents non-selectively. Thus, our findings directly demonstrate that Ca(V)3.2 channels are the main molecular substrate for redox regulation of neuronal T-type channels. In addition, because thalamic T-type channels generate low-threshold Ca(2+) spikes that directly correlate with burst firing in these neurons, differential redox regulation of these channels may have an important function in controlling cellular excitability in physiological and pathological conditions and fine-tuning of the flow of sensory information into the central nervous system.

  16. Achromatopsia as a potential candidate for gene therapy.

    PubMed

    Pang, Ji-Jing; Alexander, John; Lei, Bo; Deng, Wentao; Zhang, Keqing; Li, Qiuhong; Chang, Bo; Hauswirth, William W

    2010-01-01

    Achromatopsia is an autosomal recessive retinal disease involving loss of cone function that afflicts approximately 1 in 30,000 individuals. Patients with achromatopsia usually have visual acuities lower than 20/200 because of the central vision loss, photophobia, complete color blindness and reduced cone-mediated electroretinographic (ERG) amplitudes. Mutations in three genes have been found to be the primary causes of achromatopsia, including CNGB3 (beta subunit of the cone cyclic nucleotide-gated cation channel), CNGA3 (alpha subunit of the cone cyclic nucleotide-gated cation channel), and GNAT2 (cone specific alpha subunit of transducin). Naturally occurring mouse models with mutations in Cnga3 (cpfl5 mice) and Gnat2 (cpfl3 mice) were discovered at The Jackson Laboratory. A natural occurring canine model with CNGB3 mutations has also been found. These animal models have many of the central phenotypic features of the corresponding human diseases. Using adeno-associated virus (AAV)-mediated gene therapy, we and others show that cone function can be restored in all three models. These data suggest that human achromatopsia may be a good candidate for corrective gene therapy.

  17. Durangite from the Black Range, New Mexico, and new data on durangite from Durango and Cornwall.

    USGS Publications Warehouse

    Foord, E.E.; Oakman, M.R.; Maxwell, C.H.

    1985-01-01

    Durangite, associated with cassiterite, hematite, quartz, tridymite, cristobalite and clinopyroxene, occurs in small veinlets within flows, ash-flow tuffs and lithic tuffs in a tin mine near Boiler Peak, New Mexico. It is clear to semi-translucent, pale yellow-orange to medium orange-red with a vitreous lustre, pale yellow streak; H. 5-5.5%; irregular to conchoidal fracture and a good (110) cleavage; elongate along c with (110), (010), (021) and (111) the prominent forms; Dmeas 3.90, Dcalc 3.92 g/cm3; alpha medium yellow orange 1.634(3), beta pale yellow orange 1.663(3), gamma colourless 1.685(3); weak to moderate dispersion r < v. The structural formula is: (Na0.93Li0.07)SIGMA 1.00(Al0.89Fe0.07Mn0.06)SIGMA 1.02As0.99O4(F0.90(OH)0.07)SIGMA 0.97. Indexed XRD powder data are tabulated; a 6.574(1), b 8.505(2), c 7.019(1) A, beta 115.34o; space group C2/c; Z = 4. Additional X-ray and chemical data on durangite from Durango and Cornwall are also included.-L.T.T.

  18. Triple tracks in CR-39 as the result of Pd-D Co-deposition: evidence of energetic neutrons.

    PubMed

    Mosier-Boss, Pamela A; Szpak, Stanislaw; Gordon, Frank E; Forsley, Lawrence P G

    2009-01-01

    Since the announcement by Fleischmann and Pons that the excess enthalpy generated in the negatively polarized Pd-D-D(2)O system was attributable to nuclear reactions occurring inside the Pd lattice, there have been reports of other manifestations of nuclear activities in this system. In particular, there have been reports of tritium and helium-4 production; emission of energetic particles, gamma or X-rays, and neutrons; as well as the transmutation of elements. In this communication, the results of Pd-D co-deposition experiments conducted with the cathode in close contact with CR-39, a solid-state nuclear etch detector, are reported. Among the solitary tracks due to individual energetic particles, triple tracks are observed. Microscopic examination of the bottom of the triple track pit shows that the three lobes of the track are splitting apart from a center point. The presence of three alpha-particle tracks outgoing from a single point is diagnostic of the (12)C(n,n')3alpha carbon breakup reaction and suggests that DT reactions that produce > or = 9.6 MeV neutrons are occurring inside the Pd lattice. To our knowledge, this is the first report of the production of energetic (> or = 9.6 MeV) neutrons in the Pd-D system.

  19. Headspace volatile composition of the flowers of Caralluma europaea N.E.Br. (Apocynaceae).

    PubMed

    Formisano, Carmen; Senatore, Felice; Della Porta, Giovanna; Scognamiglio, Mariarosa; Bruno, Maurizio; Maggio, Antonella; Rosselli, Sergio; Zito, Pietro; Sajeva, Maurizio

    2009-11-11

    The volatile constituents of the flowers of Caralluma europaea (Guss.) N.E.Br (Apocynaceae) from Lampedusa Island were analyzed by a headspace GC method. The analyses allowed the identification and quantification of 41 compounds. The main components were, among the monoterpenoids, terpinolene (23.3%), alpha-terpinene (19.1%) and linalool (18.4%), whereas, among the carbonylic compounds the major constituents were heptanal (2.0%), octanoic acid (2.4%) and hexanoic acid (1.7%). The presence of a nitrogen containing compound, indole (0.8%) and of a sulphur containing compound, dimethylsulphide (t), noteworthy. The compounds found in the flowers of C. europaea have been compared with data available in the literature as regard to their odor, presence in other sapromyiophilous taxa, possible role as semiochemicals, and presence in decaying organic matter. 89.3% of total constituents have been described in other sapromyiophilous taxa. Some of the compounds are present in several types of decaying organic matter (excrements, decomposing bodies, and spoiled fish, etc). Several volatiles found in C. europaea flowers are used as semiochemicals by Hymenoptera, Coleoptera, Diptera, Lepidoptera and other insects. Sixteen volatiles, accounting for 32.4% of the total constituents, are described as attractants of some Diptera families, with a biology linked to decaying organic matter. Our data thus confirm that C. europaea floral bouquet falls within the sapromyiophilous pollination syndrome.

  20. Higher order Larmor radius corrections to guiding-centre equations and application to fast ion equilibrium distributions

    NASA Astrophysics Data System (ADS)

    Lanthaler, S.; Pfefferlé, D.; Graves, J. P.; Cooper, W. A.

    2017-04-01

    An improved set of guiding-centre equations, expanded to one order higher in Larmor radius than usually written for guiding-centre codes, are derived for curvilinear flux coordinates and implemented into the orbit following code VENUS-LEVIS. Aside from greatly improving the correspondence between guiding-centre and full particle trajectories, the most important effect of the additional Larmor radius corrections is to modify the definition of the guiding-centre’s parallel velocity via the so-called Baños drift. The correct treatment of the guiding-centre push-forward with the Baños term leads to an anisotropic shift in the phase-space distribution of guiding-centres, consistent with the well-known magnetization term. The consequence of these higher order terms are quantified in three cases where energetic ions are usually followed with standard guiding-centre equations: (1) neutral beam injection in a MAST-like low aspect-ratio spherical equilibrium where the fast ion driven current is significantly larger with respect to previous calculations, (2) fast ion losses due to resonant magnetic perturbations where a lower lost fraction and a better confinement is confirmed, (3) alpha particles in the ripple field of the European DEMO where the effect is found to be marginal.