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Selected AbstractsUltrastructure of spermiogenesis and mature spermatozoon of Anonchotaenia globata (von Linstow, 1879) (Cestoda, Cyclophyllidea, Paruterinidae)ACTA ZOOLOGICA, Issue 2 2010Aneta Yoneva Abstract Yoneva, A., Georgieva, K., Mizinska, Y., Nikolov, P. N., Georgiev, B. B. and Stoitsova, S. R. 2010. Ultrastructure of spermiogenesis and mature spermatozoon of Anonchotaenia globata (von Linstow, 1879) (Cestoda, Cyclophyllidea, Paruterinidae). , Acta Zoologica (Stockholm) 91: 184,192 The ultrastructure of spermiogenesis and of the spermatozoon of a species of the family Paruterinidae is described for the first time. The spermiogenesis of Anonchotaenia globata starts with the formation of a differentiation zone with two centrioles associated with thin striated roots. One of the centrioles gives rise to a free flagellum followed by a slight flagellar rotation and a proximodistal fusion of the flagellum with the cytoplasmic protrusion. This pattern corresponds to Type III spermiogenesis in cestodes. The spermatozoon consists of five distinct regions. The anterior extremity possesses an apical cone and a single helically coiled crested body. The cortical microtubules are spirally arranged. The axoneme is surrounded by a periaxonemal sheath and a thin layer of cytoplasm filled with electron-dense granules in Regions I,V. The periaxonemal sheath is connected with the peripheral microtubules by transverse intracytoplasmic walls in Regions III and IV. The nucleus is spirally coiled around the axoneme. Anonchotaenia globata differs from Dilepididae (where paruterinids have previously been classified) in the type of spermiogenesis, the lack of glycogen inclusions and the presence of intracytoplasmic walls. The pattern of spermiogenesis is similar to that in Metadilepididae and Taeniidae, which are considered phylogenetically close to Paruterinidae. [source] Coordinated Development of Yeast Colonies: An Experimental Analysis of the Adaptation to Different Nutrient Concentrations , Part 1ENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 2 2005T. Walther Abstract The development of yeast colonies on solid agar substrates served as a model system to investigate the growth of higher fungi in a heterogeneous environment. Applying a new analytical technique , which was based on the estimation of the intensity of transmitted light from microscopic images taken along the colony radius , cell-density distributions inside fungal mycelia were measured at an extremely high spatial resolution. Using this method, the adaptation of yeast colonies to the limitation of different nutrients was investigated. Under conditions of carbon or nitrogen limitation, populations of the dimorphic model yeasts Yarrowia lipolytica and Candida boidinii underwent a transition in their morphology from solid colony to mycelial colony patterns. When grown under conditions that induced the mycelial morphology, colonies extended linearly at a constant rate irrespective of the initial nutrient concentration. In general, the cell density within the population declined at higher degrees of limitation. Nitrogen-limited colonies of both model yeasts, as well as carbon-limited Y.,lipolytica colonies proceeded to extend until the growth field was finally covered by the population. Under these conditions, areas of fairly constant cell densities were formed during the growth process. Only carbon-limited C.,boidinii colonies stopped extending at a final diameter which was small when compared to the size of the growth field, and formed a cell-density profile which was monotonically declining. The observed differences in the final colony diameter, and in the cell-density profile morphology indicated the presence of different regulatory mechanisms that ruled the colony development of the model yeasts. The presented monitoring technique for the biomass distribution inside fungal populations provided the basis for a quantitative and non-invasive description of mycelial development. [source] Broad T cell immunity to the LcrV virulence protein is induced by targeted delivery to DEC-205/CD205-positive mouse dendritic cellsEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 1 2008Yoonkyung Do Abstract There is a need for a more efficient vaccine against the bacterium Yersinia pestis, the agent of pneumonic plague. The F1-LcrV (F1-V) subunit vaccine in alhydrogel is known to induce humoral immunity. In this study, we utilized DC to investigate cellular immunity. We genetically engineered the LcrV virulence protein into the anti-DEC-205/CD205 mAb and thereby targeted the conjugated protein directly to mouse DEC-205+ DC in situ. We observed antigen-specific CD4+ T cell immunity measured by intracellular staining for IFN-, in three different mouse strains (C57BL/6, BALB/c, and C3H/HeJ), while we could not observe such T cell responses with F1-V vaccine in alhydrogel. Using a peptide library for LcrV protein, we identified two or more distinct CD4+ T cell mimetopes in each MHC haplotype, consistent with the induction of broad immunity. When compared to nontargeted standard protein vaccine, DC targeting greatly increased the efficiency for inducing IFN-,-producing T cells. The targeted LcrV protein induced antibody responses to a similar extent as the F1-V subunit vaccine, but Th1-dependent IgG2a and IgG2c isotypes were observed only after anti-DEC-205:LcrV mAb immunization. This study sets the stage for the analysis of functional roles of IFN-,-producing T cells in Y.,pestis infection. [source] Structure Comparison of Early and Late Lanthanide(III) Homodinuclear Macrocyclic Complexes with the Polyamine Polycarboxylic Ligand H8OHECEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 19 2004Ulrike A. Böttger Abstract The solid-state structures of two new homodinuclear chelate complexes with the late lanthanide(III) ions Yb and Lu, [Na2(Yb2OHEC)].14.5H2O (1), and [Na2(Lu2OHEC)].14.5H2O (2) (H8OHEC = 1,4,7,10,14,17,20,23-octaazacyclohexacosane- 1,4,7,10,14,17,20,23-octaacetic acid), have been determined by X-ray crystal structure analysis. Each lanthanide(III) ion is coordinated by eight donor atoms of the ligand and the geometry of the coordination polyhedron approaches a bicapped trigonal prism. These structures are compared with those of the homodinuclear chelate complexes with the same ligand and the mid to early lanthanide(III) ions Gd, Eu, La and also Y. A distinctive structural change occurs across the lanthanide series. The centrosymmetric mid to early lanthanide(III) complexes are all ninefold-coordinated in a capped square antiprismatic arrangement with a water molecule coordinated in a prismatic position. This structure is maintained in aqueous solution, together with an asymmetric minor isomer. The late lanthanide(III) OHEC complexes not only lack the inner-sphere water, but the change of coordination sphere also results in a loss of symmetry of the whole complex molecule. The observed change of coordination mode and number of the lanthanide ion may offer a geometric model for the isomerization process in eight- and ninefold-coordinated complex species that are isomers in a possible coordination equilibrium observed by NMR in aqueous solution. This model may also explain the intramolecular rearrangements necessary during water exchange in the inner coordination sphere of the complex [(Gd2OHEC)(H2O)2]2, through a slow dissociative mechanism. Protonation constants of the H8OHEC ligand and complex formation constants of this ligand with GdIII, CaII, CuII and ZnII have been determined by solution thermodynamic studies. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source] The SDF-1/CXCR4 pathway and the development of the cerebellar systemEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 8 2005Tim O. Vilz Abstract Mice deficient for the chemokine receptor CXCR4 show premature translocation of granule cell neuroblasts from their germinal zone into the nascent cerebellum [Y.-R. Zuo et al. (1998)Nature, 393, 595,599]. Here, we used CXCR4-null mice to analyse the early development of cerebellar cortical inhibitory interneurons and pontine neurons which, in the adult, are synaptically integrated with granule cells. Cortical inhibitory interneuronal precursors normally invade the cerebellar anlage of CXCR4-deficient mice, but their dispersal is impeded by dislocated foci of proliferating granule cells, from which they are excluded. This is reminiscent of the strict exclusion of inhibitory interneuronal precursors from the superficial external granule cell layer. As inhibitory interneuronal precursors readily mingle with post-mitotic granule cells both in wild-type and CXCR4-null mice, these findings indicate that the developmentally regulated interactions between granule and inhibitory interneuronal precursors are independent of SDF-1/CXCR4 signalling. In contrast, the transit of pontine neurons from the rhombic lip through the anterior extramural stream to the basilar pons is disrupted in CXCR4-deficient animals. Migrating pontine neurons express CXCR4, and in CXCR4-null animals these cells are found displaced deep into the brainstem. Consequently, nascent pontine nuclei in CXCR4-deficient animals are hypoplastic. Moreover, they fail to express plexin D1, suggesting that SDF-1/CXCR4 signalling may also impinge on axon guidance critical to the orderly formation of granule cell mossy fibre afferents. [source] Humanin antagonists: mutants that interfere with dimerization inhibit neuroprotection by HumaninEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2004Yuichi Hashimoto Abstract The 24-residue peptide Humanin (HN) protects neuronal cells from insults of various Alzheimer's disease (AD) genes and A, by forming a homodimer. We have previously shown that P3A, S7A, C8A, L9A, L12A, T13A, S14A and P19A mutations nullify the neuroprotective function of HN [Yamagishi, Y., Hashimoto, Y., Niikura, T. & Nishimoto, I. (2003) Peptides, 24, 585,595]. Here we examined whether any of these ,null' mutants could function as dominant-negative mutants. Homodimerization-defective mutants, P3A-, L12A-, S14A- and P19A-HN, specifically blocked neuroprotection by HN, but not by activity-dependent neurotrophic factor. Furthermore, insertion of S7A, the mutation that blocks the homodimerization of HN, but not insertion of G5A abolished the antagonizing function of L12A-HN. While L12A-HN and G5A/L12A-HN actually inhibited HN homodimerization, S7A/L12A-HN had no effect. These data indicate that P3A-, L12A-, S14A- and P19A-HN function as HN antagonists by forming an inactive dimer with HN. This study provides a novel insight into the understanding of the in vivo function of HN, as well as into the development of clinically applicable HN neutralizers. [source] Rapid and long-term alterations of hippocampal GABAB receptors in a mouse model of temporal lobe epilepsyEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 8 2003Andrea Straessle Abstract Alterations of ,-aminobutyric acid (GABA)B receptor expression have been reported in human temporal lobe epilepsy (TLE). Here, changes in regional and cellular expression of the GABAB receptor subunits R1 (GBR1) and R2 (GBR2) were investigated in a mouse model that replicates major functional and histopathological features of TLE. Adult mice received a single, unilateral injection of kainic acid (KA) into the dorsal hippocampus, and GABAB receptor immunoreactivity was analysed between 1 day and 3 months thereafter. In control mice, GBR1 and GBR2 were distributed uniformly across the dendritic layers of CA1,CA3 and dentate gyrus. In addition, some interneurons were labelled selectively for GBR1. At 1 day post-KA, staining for both GBR1 and GBR2 was profoundly reduced in CA1, CA3c and the hilus, and no interneurons were visible anymore. At later stages, the loss of GABAB receptors persisted in CA1 and CA3, whereas staining increased gradually in dentate gyrus granule cells, which become dispersed in this model. Most strikingly, a subpopulation of strongly labelled interneurons reappeared, mainly in the hilus and CA3 starting at 1 week post-KA. In double-staining experiments, these cells were selectively labelled for neuropeptide Y. The number of GBR1-positive interneurons also increased contralaterally in the hilus. The rapid KA-induced loss of GABAB receptors might contribute to epileptogenesis because of a reduction in both presynaptic control of transmitter release and postsynaptic inhibition. In turn, the long-term increase in GABAB receptors in granule cells and specific subtypes of interneurons may represent a compensatory response to recurrent seizures. [source] Impairment of eyeblink conditioning in GluR,2-mutant mice depends on the temporal overlap between conditioned and unconditioned stimuliEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 9 2001Yasushi Kishimoto Abstract Mice lacking the glutamate receptor subunit ,2 (GluR,2) are deficient in cerebellar long-term depression (LTD) at the parallel fibre,Purkinje cell synapses. We conducted delay and trace eyeblink conditioning with these mice, using various temporal intervals between the conditioned stimulus (CS) and unconditioned stimulus (US). During trace conditioning in which a stimulus-free trace interval (TI) of 250, 100 or 50 ms intervened between the 352-ms tone CS and 100-ms US, GluR,2-mutant mice learned as successfully as wild-type mice. Even in the paradigm with TI = 0 ms, in which the end of CS and onset of US are simultaneous, there was no difference between the GluR,2-mutant and wild-type mice in their acquisition of a conditioned response. However, in the delay paradigm in which the 452-ms CS overlapped temporally with the coterminating 100-ms US, GluR,2-mutant mice exhibited severe learning impairment. The present study together with our previous work [Kishimoto, Y., Kawahara, S., Suzuki, M., Mori, H., Mishina, M. & Kirino, Y. (2001) Eur. J. Neurosci.,13, 1249,1254], indicates that cerebellar LTD-independent learning is possible in paradigms without temporal overlap between the CS and US. On the other hand, GluR,2 and cerebellar LTD are essential for learning when there is CS,US temporal overlap, suggesting that the cerebellar neural substrates underlying eyeblink conditioning may change, depending on the temporal overlap of the CS and US. [source] Pachepsky, Y., Radcliff, D.E. & Selim, H.M. (eds) Scaling Methods in Soil Physics.EUROPEAN JOURNAL OF SOIL SCIENCE, Issue 3 20042003. xviii + 434 pp., Boca Raton, CRC Press No abstract is available for this article. [source] Inactivation of colicin Y by intramembrane helix,helix interaction with its immunity proteinFEBS JOURNAL, Issue 21 2008David, majs The construction of hybrids between colicins U and Y and the mutagenesis of the colicin Y gene (cya) have revealed amino acid residues important for interactions between colicin Y and its cognate immunity protein (Cyi). Four such residues (I578, T582, Y586 and V590) were found in helices 8 and 9 of the colicin Y pore-forming domain. To verify the importance of these residues, the corresponding amino acids in the colicin B protein were mutated to the residues present in colicin Y. An Escherichia coli strain with cloned colicin Y immunity gene (cyi) inactivated this mutant, but not the wild-type colicin B. In addition, interacting amino acid pairs in Cya and Cyi were identified using a set of Cyi point mutant strains. These data are consistent with antiparallel helix,helix interactions between Cyi helix T3 and Cya helix 8 of the pore-forming domain as a molecular mechanism of colicin Y inactivation by its immunity protein. [source] Functional analysis of the basic helix-loop-helix transcription factor DEC1 in circadian regulationFEBS JOURNAL, Issue 22 2004Interaction with BMAL The basic helix-loop-helix transcription factor DEC1 is expressed in a circadian manner in the suprachiasmatic nucleus where it seems to play a role in regulating the mammalian circadian rhythm by suppressing the CLOCK/BMAL1-activated promoter. The interaction of DEC1 with BMAL1 has been suggested as one of the molecular mechanisms of the suppression [Honma, S., Kawamoto, T., Takagi, Y., Fujimoto, K., Sato, F., Noshiro, M., Kato, Y. & Honma, K. (2002) Nature419, 841,844]. Deletion analysis of DEC1 demonstrated that its N-terminal region, which includes the basic helix-loop-helix domain, was essential for both the suppressive activity and the interaction with BMAL1, as DEC1 lacking the basic region did not show any suppression or interaction. Furthermore, we found that Arg65 in the basic region, which is conserved among group B basic helix-loop-helix proteins, was responsible for the suppression, for the interaction with BMAL1 and for its binding to CACGTG E-boxes. However, substitution of His57 for Ala significantly reduced the E-box binding activity of DEC1, although it did not affect the interaction with BMAL1 or suppression of CLOCK/BMAL1-induced transcription. On the other hand, the basic region-deleted DEC1 acted in a dominant-negative manner for DEC1 activity, indicating that the basic region was not required for homodimer formation of DEC1. Moreover, mutant DEC1 also counteracted DEC2-mediated suppressive activity in a dominant-negative manner. The heterodimer formation of DEC1 and DEC2 was confirmed by pull-down assay. These findings suggest that the basic region of DEC1 participates in the transcriptional regulation through a protein,protein interaction with BMAL1 and DNA binding to the E-box. [source] The activation of gelsolin by low pHFEBS JOURNAL, Issue 20 2003The calcium latch is sensitive to calcium but not pH Gelsolin is a multidomain and multifunction protein that nucleates the assembly of filaments and severs them. The activation of gelsolin by calcium is a multistep process involving many calcium binding sites that act to unfold the molecule from a tight structure to a more loose form in which three actin-binding sites become exposed. Low pH is also known to activate gelsolin, in the absence of calcium and this too results in an unfolding of the molecule. Less is known how pH-activation occurs but we show that there are significant differences in the mechanisms that lead to activation. Crucially, while it is known that the bonds between G2 and G6 are broken by co-operative occupancy of calcium binding sites in both domains [Lagarrique, E., Maciver, S. K., Fattoum, A., Benyamin, Y. & Roustan, C. (2003) Eur. J. Biochem. 270, 2236,2243.], pH values that activate gelsolin do not result in a weakening of the G2-G6 bonds. We report the existence of pH-dependent conformational changes within G2 and in G4,6 that differ from those induced by calcium, and that low pH overrides the requirement for calcium for actin-binding within G4,6 to a modest extent so that a Kd of 1 µm is measured, compared to 30,40 nm in the presence of calcium. Whereas the pH-dependent conformational change in G2 is possibly different from the change induced by calcium, the changes measured in G4,6 appear to be similar in both calcium and low pH. [source] Proteasome-driven turnover of tryptophan hydroxylase is triggered by phosphorylation in RBL2H3 cells, a serotonin producing mast cell lineFEBS JOURNAL, Issue 19 2002Yoshiko Iida We previously demonstrated in mast cell lines RBL2H3 and FMA3 that tryptophan hydroxylase (TPH) undergoes very fast turnover driven by 26S-proteasomes [Kojima, M., Oguro, K., Sawabe, K., Iida, Y., Ikeda, R., Yamashita, A., Nakanishi, N. & Hasegawa, H. (2000) J. Biochem (Tokyo) 2000, 127, 121,127]. In the present study, we have examined an involvement of TPH phosphorylation in the rapid turnover, using non-neural TPH. The proteasome-driven degradation of TPH in living cells was accelerated by okadaic acid, a protein phosphatase inhibitor. Incorporation of 32P into a 53-kDa protein, which was judged to be TPH based on autoradiography and Western blot analysis using anti-TPH serum and purified TPH as the size marker, was observed in FMA3 cells only in the presence of both okadaic acid and MG132, inhibitors of protein phosphatase and proteasome, respectively. In a cell-free proteasome system constituted mainly of RBL2H3 cell extracts, degradation of exogenous TPH isolated from mastocytoma P-815 cells was inhibited by protein kinase inhibitors KN-62 and K252a but not by H89. Consistent with the inhibitor specificity, the same TPH was phosphorylated by exogenous Ca2+/calmodulin-dependent protein kinase II in the presence of Ca2+ and calmodulin but not by protein kinase A (catalytic subunit). TPH protein thus phosphorylated by Ca2+/calmodulin-dependent protein kinase II was digested more rapidly in the cell-free proteasome system than was the nonphosphorylated enzyme. These results indicated that the phosphorylation of TPH was a prerequisite for proteasome-driven TPH degradation. [source] Casein kinase 2 specifically binds to and phosphorylates the carboxy termini of ENaC subunitsFEBS JOURNAL, Issue 18 2002Haikun Shi A number of findings have suggested the involvement of protein phosphorylation in the regulation of the epithelial Na+ channel (ENaC). A recent study has demonstrated that the C tails of the , and , subunits of ENaC are subject to phosphorylation by at least three protein kinases [Shi, H., Asher, C., Chigaev, A., Yung, Y., Reuveny, E., Seger, R. & Garty, H. (2002) J. Biol. Chem. 277, 13539,13547]. One of them was identified as ERK which phosphorylates ,T613 and ,T623 and affects the channel interaction with Nedd4. The current study identifies a second protein kinase as casein kinase 2 (CK2), or CK-2-like kinase. It phosphorylates ,S631, a well-conserved serine on the , subunit. Such phosphorylation is observed both in vitro using glutathione-S-transferase,ENaC fusion proteins and in vivo in ENaC-expressing Xenopus oocytes. The , subunit is weakly phosphorylated by this protein kinase on another residue (,T599), and the C tail of , is not significantly phosphorylated by this kinase. Thus, CK2 may be involved in the regulation of the epithelial Na+ channel. [source] Interleukin-6-induced proliferation of pre-B cells mediated by receptor complexes lacking the SHP2/SOCS3 recruitment sites revisitedFEBS JOURNAL, Issue 24 2001Kerstin Friederichs Interleukin-6 (IL-6) induces B-cell proliferation by binding to receptor complexes composed of a specific ,-receptor (gp80; CD126) and the signal transducing receptor subunit gp130 (CD130). Immediately after receptor complex activation, signal transducers and activators of transcription (STATs) 1 and 3 and the Src-homology domain-containing protein tyrosine phosphatase 2 (SHP2) are recruited to gp130 and subsequently tyrosine phosphorylated. The activated dimerized STATs translocate to the nucleus and bind to enhancer elements of IL-6-inducible genes. SHP2 acts as an adapter and links the Jak/STAT pathway to the Ras/Raf/MAPK cascade but it is also involved in signal attenuation. Whereas STAT3 activation appears to be crucial for all biological activities of IL-6, the requirement of SHP2-activation depends on the individual biological response analyzed. The requirement of SHP2 activation for the pre-B cell (Ba/F3) proliferation has been reported previously [Fukada, T., Hibi, M., Yamanaka, Y., Takahashi-Tezuka, M., Fujitani, Y., Yamaguchi, T., Nakajima, K. & Hirano, T. (1996) Immunity5, 449,460]. In contrast, we have recently demonstrated that the presence of a single STAT-recruitment site within gp130 is sufficient for IL-6- induced proliferation of Ba/F3 cells [Schmitz, J., Dahmen, H., Grimm, C., Gendo, C., Müller-Newen, G., Heinrich, P.C. & Schaper, F. (2000) J. Immunol.164, 848,854]. To unravel this discrepancy we analyzed the IL-6-induced dose-dependent proliferation of Ba/F3 cells mediated by receptor complexes lacking SHP2/SOCS3 recruitment sites. Surprisingly, pre-B cells, after stimulation with low amounts of IL-6, proliferate much more efficiently in the absence of the activated SHP2 than in the presence of the tyrosine phosphatase. Therefore, SHP2 activation appears to be relevant for IL-6-induced proliferation only after stimulation with very large amounts of IL-6. [source] E2F1-mediated transcriptional inhibition of the plasminogen activator inhibitor type 1 geneFEBS JOURNAL, Issue 18 2001Magdalena Koziczak ,Gene expression of the plasminogen activation system is cell-cycle dependent. Previously, we showed that ectopic expression of E2F1 repressed the plasminogen activator inhibitor type 1 (PAI-1) promoter in a manner dependent on the presence of DNA-binding and transactivation domains of E2F1 but independent of binding to pocket-binding proteins, suggesting a novel mechanism for E2F-mediated negative gene regulation [Koziczak, M., Krek, W. & Nagamine, Y. (2000) Mol. Cell. Biol.20, 2014,2022]. However, it remains to be seen whether endogenous E2F can exert a similar effect. We report here that down-regulation of PAI-1 gene expression correlates with an increase in endogenous E2F activity. When cells were treated with a cdk2/4-specific inhibitor, which maintains E2F in an inactive state, the decline of serum-induced PAI-1 mRNA levels was suppressed. In mutant U2OS cells expressing a temperature-sensitive retinoblastoma protein (pRB), a shift to a permissive temperature induced PAI-1 mRNA expression. In U2OS cells stably expressing an E2F1-estrogen receptor chimeric protein that could be activated by tamoxifen, PAI-1 gene transcription was markedly reduced by tamoxifen even in the presence of cycloheximide. These results all indicate that endogenous E2F can directly repress the PAI-1 gene. DNase I hypersensitive-site analysis of the PAI-1 promoter suggested the involvement of conformation changes in chromatin structure of the PAI-1 promoter. 5, deletion analysis of the PAI-1 promoter showed that multiple sites were responsible for the E2F negative regulation, some of which were promoter dependent. Interestingly, one of these sites is a p53-binding element. [source] Differential actions of p60c-Src and Lck kinases on the Ras regulators p120-GAP and GDP/GTP exchange factor CDC25MmFEBS JOURNAL, Issue 11 2001Carmela Giglione It is known that the human Ras GTPase activating protein (GAP) p120-GAP can be phosphorylated by different members of the Src kinase family and recently phosphorylation of the GDP/GTP exchange factor (GEF) CDC25Mm/GRF1 by proteins of the Src kinase family has been revealed in vivo[Kiyono, M., Kaziro, Y. & Satoh, T. (2000) J. Biol. Chem.275, 5441,5446]. As it still remains unclear how these phosphorylations can influence the Ras pathway we have analyzed the ability of p60c-Src and Lck to phosphorylate these two Ras regulators and have compared the activity of the phosphorylated and unphosphorylated forms. Both kinases were found to phosphorylate full-length or truncated forms of GAP and GEF. The use of the catalytic domain of p60c-Src showed that its SH3/SH2 domains are not required for the interaction and the phosphorylation of both regulators. Remarkably, the phosphorylations by the two kinases were accompanied by different functional effects. The phosphorylation of p120-GAP by p60c-Src inhibited its ability to stimulate the Ha-Ras-GTPase activity, whereas phosphorylation by Lck did not display any effect. A different picture became evident with CDC25Mm; phosphorylation by Lck increased its capacity to stimulate the GDP/GTP exchange on Ha-Ras, whereas its phosphorylation by p60c-Src was ineffective. Our results suggest that phosphorylation by p60c-Src and Lck is a selective process that can modulate the activity of p120-GAP and CDC25Mm towards Ras proteins. [source] Structure and promoter analysis of the mouse membrane-bound transferrin-like protein (MTf) geneFEBS JOURNAL, Issue 5 2001Kazuko Nakamasu Recently, we purified membrane-bound transferrin-like protein (MTf) from the plasma membrane of rabbit chondrocytes and showed that the expression levels of MTf protein and mRNA were much higher in cartilage than in other tissues [Kawamoto T, Pan, H., Yan, W., Ishida, H., Usui, E., Oda, R., Nakamasu, K., Noshiro, M., Kawashima-Ohya, Y., Fujii, M., Shintani, H., Okada, Y. & Kato, Y. (1998) Eur. J. Biochem.256, 503,509]. In this study, we isolated the MTf gene from a constructed mouse genomic library. The mouse MTf gene was encoded by a single-copy gene spanning ,,26 kb and consisting of 16 exons. The transcription-initiation site was located 157 bp upstream from the translation-start codon, and a TATA box was not found in the 5, flanking region. The mouse MTf gene was mapped on the B3 band of chromosome 16 by fluorescence in situ hybridization. Using primary chondrocytes, SK-MEL-28 (melanoma cell line), ATDC5 (chondrogenic cell line) and NIH3T3 (fibroblast cell line) cells, we carried out transient expression studies on various lengths of the 5, flanking region of the MTf gene fused to the luciferase reporter gene. Luciferase activity in SK-MEL-28 cells was higher than in primary chondrocytes. Although no luciferase activity was detectable in NIH3T3 cells, it was higher in chondrocytes than in ATDC5 chondrogenic cells. These findings were consistent with the levels of expression of MTf mRNA in these cells cultured under similar conditions. The patterns of increase and decrease in the luciferase activity in chondrocytes transfected with various 5, deleted constructs of the MTf promoter were similar to that in ATDC5 cells, but differed from that in SK-MEL-28 cells. The findings obtained with primary chondrocytes suggest that the regions between ,693 and ,444 and between ,1635 and ,1213 contain positive and negative cis -acting elements, respectively. The chondrocyte-specific expression of the MTf gene could be regulated via these regulatory elements in the promoter region. [source] Dual metabolic pathway of 25-hydroxyvitamin D3 catalyzed by human CYP24FEBS JOURNAL, Issue 20 2000Toshiyuki Sakaki Human 25-hydroxyvitamin D3 (25(OH)D3) 24-hydroxylase (CYP24) cDNA was expressed in Escherichia coli, and its enzymatic and spectral properties were revealed. The reconstituted system containing the membrane fraction prepared from recombinant E. coli cells, adrenodoxin and adrenodoxin reductase was examined for the metabolism of 25(OH)D3, 1,,25(OH)2D3 and their related compounds. Human CYP24 demonstrated a remarkable metabolism consisting of both C-23 and C-24 hydroxylation pathways towards both 25(OH)D3 and 1,,25(OH)2D3, whereas rat CYP24 showed almost no C-23 hydroxylation pathway [Sakaki, T. Sawada, N. Nonaka, Y. Ohyama, Y. & Inouye, K. (1999) Eur. J. Biochem. 262, 43,48]. HPLC analysis and mass spectrometric analysis revealed that human CYP24 catalyzed all the steps of the C-23 hydroxylation pathway from 25(OH)D3 via 23S,25(OH)2D3, 23S,25,26(OH)3D3 and 25(OH)D3 -26,23-lactol to 25(OH)D3 -26,23-lactone in addition to the C-24 hydroxylation pathway from 25(OH)D3 via 24R,25(OH)2D3, 24-oxo-25(OH)D3, 24-oxo-23S,25(OH)2D3 to 24,25,26,27-tetranor-23(OH)D3. On 1,,25(OH)2D3 metabolism, similar results were observed. These results strongly suggest that the single enzyme human CYP24 is greatly responsible for the metabolism of both 25(OH)D3 and 1,,25(OH)2D3. We also succeeded in the coexpression of CYP24, adrenodoxin and NADPH-adrenodoxin reductase in E. coli. Addition of 25(OH)D3 to the recombinant E. coli cell culture yielded most of the metabolites in both the C-23 and C-24 hydroxylation pathways. Thus, the E. coli expression system for human CYP24 appears quite useful in predicting the metabolism of vitamin D analogs used as drugs. [source] Novel polysialogangliosides of skate brainFEBS JOURNAL, Issue 16 2000Structural determination of tetra, hexasialogangliosides with a NeuAc-GalNAc linkage, penta The gangliosides in the brain of a cartilaginous fish, skate (Bathyraja smirnovi), have been isolated and characterized by means of methylation analysis, antibody binding, enzymatic hydrolysis and MALDI-TOF MS. In addition to gangliosides with known structures (GM2, fucosyl-GM1, GD3, GD2, GT3 and GT2), five polysialogangliosides were isolated and characterized as having the following structures. (1) IV3NeuAc, III6NeuAc, II3NeuAc-Gg4Cer; (2) IV3NeuAc2, III6NeuAc, II3NeuAc-Gg4Cer; (3) IV3NeuAc, III6NeuAc, II3NeuAc2 -Gg4Cer; (4) IV3NeuAc, III6NeuAc, II3NeuAc3 -Gg4Cer; and (5) IV3NeuAc2, III6NeuAc, II3NeuAc3 -Gg4Cer. These structures are ,hybrid-type' which comprise combinations of ,-series and either a, b or c-series structures. Three gangliosides (2), (4) and (5), were novel. The main features of the ganglioside composition of skate brain were an abundance of gangliotriaosyl species, a lack of gangliotetraosyl species (except fucosyl-GM1), and an abundance of hybrid-types. These characteristics closely resemble those in shark brain which we reported previously [Nakamura, K., Tamai, Y. & Kasama, T. (1997) Neurochem. Int.30, 593,604]. Two of the hybrid-type gangliosides (1) and (4), were examined for their neuritogenic activity toward cultured neuronal cells (Neuro-2A), and were found to have more potent activity than nonhybrid-type gangliosides such as GM1. [source] Fluorescent Imaging: Surface Modification of Exfoliated Layered Gadolinium Hydroxide for the Development of Multimodal Contrast Agents for MRI and Fluorescence Imaging (Adv. Funct.ADVANCED FUNCTIONAL MATERIALS, Issue 21 2009Mater. The synthesis of a new stable colloid of fluorescent LGdH layers through a newly developed surface modification method is reported by Y.-S. Yoon et al. on page 3375. This new method involves layer exfoliation, anion exchange, and PEG coating. The efficient MRI contrast-enhancement of these LGdH layers, both in vitro and in vivo, demonstrates their potential utility as a multimodal probe combining optical and MR imaging. [source] Teaching and Learning Guide for: The Geopolitics of Climate ChangeGEOGRAPHY COMPASS (ELECTRONIC), Issue 5 2008Jon Barnett Author's Introduction Climate change is a security problem in as much as the kinds of environmental changes that may result pose risks to peace and development. However, responsibilities for the causes of climate change, vulnerability to its effects, and capacity to solve the problem, are not equally distributed between countries, classes and cultures. There is no uniformity in the geopolitics of climate change, and this impedes solutions. Author Recommends 1.,Adger, W. N., et al. (eds) (2006). Fairness in adaptation to climate change. Cambridge, MA: MIT Press. A comprehensive collection of articles on the justice dimensions of adaptation to climate change. Chapters discuss potential points at which climate change becomes ,dangerous', the issue of adaptation under the United Nations Framework Convention on Climate Change (UNFCCC), the unequal outcomes of adaptation within a society, the effects of violent conflict on adaptation, the costs of adaptation, and examples from Bangladesh, Tanzania, Botswana, and Hungary. 2.,Leichenko, R., and O'Brien, K. (2008). Environmental change and globalization: double exposures. New York: Oxford University Press. This book uses examples from around the world to show the way global economic and political processes interact with environmental changes to create unequal outcomes within and across societies. A very clear demonstration of the way vulnerability to environmental change is as much driven by social processes as environmental ones, and how solutions lie within the realm of decisions about ,development' and ,environment'. 3.,Nordås, R., and Gleditsch, N. (2007). Climate conflict: common sense or nonsense? Political Geography 26 (6), pp. 627,638. doi:10.1016/j.polgeo.2007.06.003 An up-to-date, systematic and balanced review of research on the links between climate change and violent conflict. See also the other papers in this special issue of Political Geography. 4.,Parry, M., et al. (eds) (2007). Climate change 2007: impacts adaptation and vulnerability. Contribution of Working Group II to the fourth assessment report of the intergovernmental panel on climate change. Cambridge, UK: Cambridge University Press. The definitive review of all the peer-reviewed research on the way climate change may impact on places and sectors across the world. Includes chapters on ecosystems, health, human settlements, primary industries, water resources, and the major regions of the world. All chapters are available online at http://www.ipcc.ch/ipccreports/ar4-wg2.htm 5.,Salehyan, I. (2008). From climate change to conflict? No consensus yet. Journal of Peace Research 45 (3), pp. 315,326. doi:10.1177/0022343308088812 A balanced review of research on the links between climate change and conflict, with attention to existing evidence. 6.,Schwartz, P., and Randall, D. (2003). An abrupt climate change scenario and its implications for United States national security. San Francisco, CA: Global Business Network. Gives insight into how the US security policy community is framing the problem of climate change. This needs to be read critically. Available at http://www.gbn.com/ArticleDisplayServlet.srv?aid=26231 7.,German Advisory Council on Global Change. (2007). World in transition: climate change as a security risk. Berlin, Germany: WBGU. A major report from the German Advisory Council on Global Change on the risks climate changes poses to peace and stability. Needs to be read with caution. Summary and background studies are available online at http://www.wbgu.de/wbgu_jg2007_engl.html 8.,Yamin, F., and Depedge, J. (2004). The International climate change regime: a guide to rules, institutions and procedures. Cambridge, UK: Cambridge University Press. A clear and very detailed explanation of the UNFCCC's objectives, actors, history, and challenges. A must read for anyone seeking to understand the UNFCCC process, written by two scholars with practical experience in negotiations. Online Materials 1.,Environmental Change and Security Program at the Woodrow Wilson International Center for Scholars http://www.wilsoncenter.org/ecsp The major website for information about environmental security. From here, you can download many reports and studies, including the Environmental Change and Security Project Report. 2.,Global Environmental Change and Human Security Project http://www.gechs.org This website is a clearing house for work and events on environmental change and human security. 3.,Intergovernmental Panel on Climate Change (IPCC) http://www.ipcc.ch/ From this website, you can download all the chapters of all the IPCC's reports, including its comprehensive and highly influential assessment reports, the most recent of which was published in 2007. The IPCC were awarded of the Nobel Peace Prize ,for their efforts to build up and disseminate greater knowledge about man-made (sic) climate change, and to lay the foundations for the measures that are needed to counteract such change'. 4.,Tyndall Centre for Climate Change Research http://www.tyndall.ac.uk The website of a major centre for research on climate change, and probably the world's leading centre for social science based analysis of climate change. From this site, you can download many publications about mitigation of and adaptation to climate change, and about various issues in the UNFCCC. 5.,United Nations Framework Convention on Climate Change http://unfccc.int/ The website contains every major document relation to the UNFCCC and its Kyoto Protocol, including the text of the agreements, national communications, country submissions, negotiated outcomes, and background documents about most key issues. Sample Syllabus: The Geopolitics of Climate Change topics for lecture and discussion Week I: Introduction Barnett, J. (2007). The geopolitics of climate change. Geography Compass 1 (6), pp. 1361,1375. United Nations Secretary General, Kofi Annan, address to the 12th Conference of Parties to the United Nations Framework Convention on Climate Change, Nairobi, 15 November 2006. Available online at http://www.unep.org/Documents.Multilingual/Default.asp?DocumentID=495&ArticleID=5424&l=en Week II: The History and Geography of Greenhouse Gas Emissions Topic: The drivers of climate change in space and time Reading Baer, P. (2006). Adaptation: who pays whom? In: Adger, N., et al. (eds) Fairness in adaptation to climate change. Cambridge, MA: MIT Press, pp. 131,154. Boyden, S., and Dovers, S. (1992). Natural-resource consumption and its environmental impacts in the Western World: impacts of increasing per capita consumption. Ambio 21 (1), pp. 63,69. Week III: The Environmental Consequences of climate change Topic: The risks climate change poses to environmental systems Reading Intergovernmental Panel on Climate Change. (2007). Climate change 2007: climate change impacts, adaptation and vulnerability: summary for policymakers. Geneva, Switzerland: IPCC Secretariat. Watch: Al Gore. The Inconvenient Truth. Weeks IV and V: The Social Consequences of Climate Change Topic: The risks climate change poses to social systems Reading Adger, W. N. (1999). Social vulnerability to climate change and extremes in coastal Vietnam. World Development 27, pp. 249,269. Comrie, A. (2007). Climate change and human health. Geography Compass 1 (3), pp. 325,339. Leary, N., et al. (2006). For whom the bell tolls: vulnerability in a changing climate. A Synthesis from the AIACC project, AIACC Working Paper No. 21, International START Secretariat, Florida. Stern, N. (2007). Economics of climate change: the Stern review. Cambridge, UK: Cambridge University Press (Chapters 3,5). Week VI: Mitigation of Climate Change: The UNFCCC Topic: The UNFCCC and the Kyoto Protocol Reading Najam, A., Huq, S., and Sokona, Y. (2003). Climate negotiations beyond Kyoto: developing countries concerns and interests. Climate Policy 3 (3), pp. 221,231. UNFCCC Secretariat. (2005). Caring for climate: a guide to the climate change convention and the Kyoto Protocol. Bonn, Germany: UN Framework Convention on Climate Change Secretariat. Weeks VII and VIII: Adaptation to Climate Change Topic: What can be done to allow societies to adapt to avoid climate impacts? Reading Adger, N., et al. (2007). Assessment of adaptation practices, options, constraints and capacity. In: Parry, M., et al. (eds) Climate change 2007: impacts, adaptation and vulnerability. Contribution of Working Group II to the fourth assessment report of the intergovernmental panel on climate change. Cambridge, UK: Cambridge University Press, pp. 717,744. Burton, I., et al. (2002). From impacts assessment to adaptation priorities: the shaping of adaptation policy. Climate Policy 2 (2,3), pp. 145,159. Eakin, H., and Lemos, M. C. (2006). Adaptation and the state: Latin America and the challenge of capacity-building under globalization. Global Environmental Change: Human and Policy Dimensions 16 (1), pp. 7,18. Ziervogel, G., Bharwani, S., and Downing, T. (2006). Adapting to climate variability: pumpkins, people and policy. Natural Resources Forum 30, pp. 294,305. Weeks IX and X: Climate Change and Migration Topic: Will climate change force migration? Readings Gaim, K. (1997). Environmental causes and impact of refugee movements: a critique of the current debate. Disasters 21 (1), pp. 20,38. McLeman, R., and Smit, B. (2006). Migration as adaptation to climate change. Climatic Change 76 (1), pp. 31,53. Myers, N. (2002). Environmental refugees: a growing phenomenon of the 21st century. Philosophical Transactions of the Royal Society 357 (1420), pp. 609,613. Perch-Nielsen, S., Bättig, M., and Imboden, D. (2008). Exploring the link between climate change and migration. Climatic Change (online first, forthcoming); doi:10.1007/s10584-008-9416-y Weeks XI and XII: Climate Change and Violent Conflict Topic: Will Climate change cause violent conflict? Readings Barnett, J., and Adger, N. (2007). Climate change, human security and violent conflict. Political Geography 26 (6), pp. 639,655. Centre for Strategic and International Studies. (2007). The age of consequences: the foreign policy and national security implications of global climate change. Washington, DC: CSIS. Nordås, R., and Gleditsch, N. (2007). Climate conflict: common sense or nonsense? Political Geography 26 (6), pp. 627,638. Schwartz, P., and Randall, D. (2003). An abrupt climate change scenario and its implications for United States national security. San Francisco, CA: Global Business Network. [online]. Retrieved on 8 April 2007 from http://www.gbn.com/ArticleDisplayServlet.srv?aid=26231 Focus Questions 1Who is most responsible for climate change? 2Who is most vulnerable to climate change? 3Does everyone have equal power in the UNFCCC process? 4Will climate change force people to migrate? Who? 5What is the relationship between adaptation to climate change and violent conflict? [source] Reply to comment on ,Sheng Y. 2001.HYDROLOGICAL PROCESSES, Issue 21 2007A bivariate gamma distribution for use in multivariate flood frequency analysis. No abstract is available for this article. [source] Inverse Monte Carlo procedure for conformation determination of macromoleculesJOURNAL OF COMPUTATIONAL CHEMISTRY, Issue 7 2003Mark Bathe Abstract A novel numerical method for determining the conformational structure of macromolecules is applied to idealized biomacromolecules in solution. The method computes effective inter-residue interaction potentials solely from the corresponding radial distribution functions, such as would be obtained from experimental data. The interaction potentials generate conformational ensembles that reproduce thermodynamic properties of the macromolecule (mean energy and heat capacity) in addition to the target radial distribution functions. As an evaluation of its utility in structure determination, we apply the method to a homopolymer and a heteropolymer model of a three-helix bundle protein [Zhou, Y.; Karplus, M. Proc Natl Acad Sci USA 1997, 94, 14429; Zhou, Y. et al. J Chem Phys 1997, 107, 10691] at various thermodynamic state points, including the ordered globule, disordered globule, and random coil states. © 2003 Wiley Periodicals, Inc. J Comput Chem 24: 876,890, 2003 [source] Protein intake, growth and lung function of infants with chronic lung diseaseJOURNAL OF HUMAN NUTRITION & DIETETICS, Issue 3 2009E. Cillié Background:, The increased survival rate of extremely preterm infants has not improved the incidence or outcome of infants diagnosed with chronic lung disease (CLD) (Riley, 2008). The relationship between optimal nutrition (particularly protein intake) and chronic lung disease has not been established. The aim of this study was to investigate the association between protein intake, growth and lung function in infants with CLD. Methods:, A CLD database, maintained for the past 10 years, was used to select participants that had reached 1 year of corrected age. Infants who were born during 2001,2006 with a birth weight of <1500 g, and who subsequently had a diagnosis of CLD, were included. Infants with evidence of intra-uterine growth restriction and abnormal cerebral pathology were excluded. Demographic, mean weight gain, protein intake and respiratory support data were collected retrospectively from the medical notes. Growth parameters and need for oxygen and inhalers up to 1 year of corrected age were collected from the CLD follow-up database. SPSS, version 15 (SPSS Inc., Chicago, IL, USA) were used for Pearson's or Spearmans correlation analysis and analysis of variance or the Wilcoxon test, as appropriate. Results:, Sixty infants were studied: 25 females and 35 males. The median (range) post-menstrual age at birth was 26 (22,31) weeks. The most common feed was breast milk; fortified breast milk was used for 37% of the total days studied. The mean (SD) protein intake was 2.28 (0.33) g kg,1 day,1 and the mean (SD) weight gain was 11.67 (1.77) g kg,1 day,1. There was a positive correlation between protein intake and weight gain (r = 0.32, P = 0.013), which was stronger in females (r = 0.51, P = 0.009). Protein intake was significantly associated with head circumference growth in females only (r = 0.47, P = 0.038). Protein intake was inversely related to the number of days spent mechanically ventilated (r = ,0.32, P = 0.015). There was no relationship between protein intake and growth at 1 year corrected age, time spent on continuous positive airway pressure, age weaned off oxygen, or the use of inhalers. There was an inverse correlation between total weeks of oxygen dependence and head circumference at 1 year (r = ,0.35, P = 0.022). Discussion:, The mean protein intake was <3 g kg,1 day,1, which is the minimum requirement for preterm infants (Tsang et al., 2005). This was associated with a sub-optimal weight gain in our participants of <15 g kg,1 day,1 (Steward & Pridham, 2002). The study demonstrates the known association between low protein intake and poor growth with ventilator dependence (Loui et al., 2008). Conclusions:, Low birth weight and low gestational age infants at risk of CLD should receive special attention to optimise their protein intake because sub-optimal protein intake potentially leads to poor growth when on a neonatal intensive care unit. References Loui, A., Tsalikaki, E., Maier, K., Walch, E., Kamarianakis, Y. & Obladen, M. (2008) Growth in high risk infants <1500 g birth weight during the first 5 weeks. Early Hum. Dev. 84, 645,650, Doi: 10.1016/j.earlhumdev.2008.04.005. Riley, K., Roth, S., Sellwood, M. & Wyatt, J.S. (2008) Survival and neurodevelopmental morbidity at 1 year of age following extremely preterm delivery over a 20-year period: a single centre cohort study. Acta Paediatr.97, 159,165. Steward, D.K. & Pridham, K.F. (2002) Growth patterns of extremely low-birth-weight hospitalised preterm infants. JOGN Nurs31, 57,65. Tsang, R.C., Uauy, R., Koletzko, B. & Zlotkin, S.H., eds. (2005) Nutrition of the Preterm Infant: Scientific Basis and Practical Guidelines. Cincinnati: Digital Educational Publishing. [source] The efficacy of dietetic intervention in patients with chronic obstructive pulmonary diseaseJOURNAL OF HUMAN NUTRITION & DIETETICS, Issue 4 2008L. Bottle Background:, Clinical trials have shown that pulmonary rehabilitation can improve the functional status and quality of life of chronic obstructive pulmonary disease (COPD) patients (Lacasse, 2006) but there is no research examining the efficacy of group dietetic intervention during standard 8 week rehabilitation courses. Current input is usually limited to a 1 h nutrition education session. This pilot study aimed to investigate whether patients receiving additional dietetic intervention during pulmonary rehabilitation significantly increased their general nutritional knowledge, thereby facilitating improvements in dietary intake and nutritional status. Methods:, Patients were recruited from two courses of pulmonary rehabilitation and randomly allocated to a control group or an intervention group. Anthropometry (height, weight, body mass index, mid arm circumference and triceps skinfold), 3 day food diaries and nutritional knowledge questionnaires covered guidelines, food groups, choosing healthy options and diet and COPD were completed at baseline and at the end of 8 weeks. In week 2 both groups received the same nutrition education session which covered healthy eating during periods of stability as well as advice on coping with loss of appetite and reduced intake during illness and exacerbations. The intervention group was followed up during weeks 4, 6 and 7 when further anthropometric measurements were taken and additional dietary advice was provided, which addressed issues raised by individual patients. Information from food diaries was converted to nutrients using Windiets dietary analysis software. Statistical analyses were carried out using SPSS (v14) and included Mann,Whitney U non parametric tests, paired t -tests and Spearman correlations used for comparisons over time and between groups. For analysis purposes patients were classified as normal weight (NW) and overweight (OW). Approval was obtained from the appropriate Ethics Committee. Results:, Changes reported were not statistically significant (P > 0.05). Complete data sets were obtained for six control (NW = 2, OW = 4) and five intervention (NW = 1, OW = 4) patients. Nutritional knowledge increased in the control group by 5% compared to 3% in the intervention group. Control NW patients increased their energy intake resulting in a mean weight gain of 0.5 kg (SD 3.3). OW control group patients increased their energy intake by 12.4% (16.9) with a mean weight gain of 0.2 kg (2.5). All control patients increased their intake of in total fat, saturated fatty acids (SFA), sugars and sodium. Conversely there was a decrease in energy intake in the intervention group of 14.4% (17.8) and a mean weight loss of 1.5 kg (1.2) (three out of four overweight patients lost weight). Improvements in diet were shown with reduced intakes of total fat, SFA, sugars and sodium. The NW patient in the intervention group regained weight that had previously been lost. These changes did not correlate with changes in nutritional knowledge. Discussion:, An increase in nutritional knowledge was expected to facilitate appropriate changes in dietary intake and nutritional status. Despite the lack of correlation between dietary knowledge and intake, beneficial outcomes were none-the-less observed in the intervention group. The trend for weight gain in OW control group patients, and weight loss in OW intervention group patients contrasted with results seen by Slinde et al. (2002) where the control OW patients lost weight, and OW intervention patients gained weight. It is possible that in the current study, patients in the intervention group were motivated to lose weight with repeated exposure to the dietitian, rather than an increase in nutritional knowledge. Significant anthropometrical changes were unlikely to be observed in 8 weeks, and further follow up may be necessary to establish sufficient evidence for the most efficacious level of dietetic intervention. The small sample sizes, especially with regard to weight sub groups, limits the conclusions which can be drawn. Further research is recommended, using a larger sample size, in order to make recommendations for dietetic best practice. Conclusion:, The results of this study did not show statistical significance and the association between nutritional knowledge and improved nutritional outcomes remains unclear. However, the findings may have clinical significance since they appear to show that additional dietetic intervention may benefit the nutritional status of patients with COPD attending pulmonary rehabilitation. References, Lacasse, Y., Goldstein, R., et al. (2006) Pulmonary rehabilitation for chronic obstructive pulmonary disease. Cochrane Database Syst. Rev. 4, CD003793. Slinde, F., Gronberg, A.M., et al. (2002) Individual dietary intervention in patients with COPD during multidisciplinary rehabilitation. Respir. Med. 96, 330,336. [source] Influence of carboxypeptidases on free amino acid, peptide and methylpyrazine contents of under-fermented cocoa beansJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 13 2002I Yusep Abstract A study of the action of two carboxypeptidases on free amino acids, peptides and methylpyrazines in under-fermented cocoa beans was carried out. Carboxypeptidase B from porcine pancreas and carboxypeptidase Y from baker's yeast were used separately for digestion. Hydrophobic free amino acids (alanine, valine, isoleucine, leucine, phenylalanine and tyrosine) were predominantly produced in samples digested with both carboxypeptidase B and Y. The peptide patterns of samples digested with both carboxypeptidases were similar to that of the control. The concentration of 2,3,5,6-tetramethylpyrazine in samples with carboxypeptidase B addition was significantly higher than those of 2,5-dimethyl- and 2,3,5-trimethylpyrazine; the concentration of 2,3,5-trimethylpyrazine was highest (1727.86,µg per 100,g) in the sample with carboxypeptidase B addition that had been incubated for 24,h. These findings indicate that carboxypeptidase B from porcine pancreas was more prominent in the formation of cocoa-specific aroma. © 2002 Society of Chemical Industry [source] Pharmacokinetic profile and behavioral effects of gabapentin in the horseJOURNAL OF VETERINARY PHARMACOLOGY & THERAPEUTICS, Issue 5 2010R. L. TERRY Terry, R. L., McDonnell, S. M., van Eps, A. W., Soma, L. R., Liu, Y., Uboh, C. E., Moate, P. J., Driessen, B. Pharmacokinetic profile and behavioral effects of gabapentin in the horse. J. vet. Pharmacol. Therap. 33, 485,494. Gabapentin is being used in horses although its pharmacokinetic (PK) profile, pharmacodynamic (PD) effects and safety in the equine are not fully investigated. Therefore, we characterized PKs and cardiovascular and behavioral effects of gabapentin in horses. Gabapentin (20 mg/kg) was administered i.v. or p.o. to six horses using a randomized crossover design. Plasma gabapentin concentrations were measured in samples collected 0,48 h postadministration employing liquid chromatography-tandem mass spectrometry. Blood pressures, ECG, and sedation scores were recorded before and for 12 h after gabapentin dosage. Nineteen quantitative measures of behaviors were evaluated. After i.v. gabapentin, the decline in plasma drug concentration over time was best described by a 3-compartment mammillary model. Terminal elimination half-life (t1/2,) was 8.5 (7.1,13.3) h. After p.o. gabapentin terminal elimination half-life () was 7.7 (6.7,11.9) h. The mean oral bioavailability of gabapentin (±SD) was 16.2 ± 2.8% indicating relatively poor absorption of gabapentin following oral administration in horses. Gabapentin caused a significant increase in sedation scores for 1 h after i.v. dose only (P < 0.05). Among behaviors, drinking frequency was greater and standing rest duration was lower with i.v. gabapentin (P < 0.05). Horses tolerated both i.v. and p.o. gabapentin doses well. There were no significant differences in and . Oral administration yielded much lower plasma concentrations because of low bioavailability. [source] Eigenshape analysis of ammonoid suturesLETHAIA, Issue 2 2010TAKAO UBUKATA Ubukata, T., Tanabe, K., Shigeta, Y., Maeda, H. & Mapes, R.H. 2009: Eigenshape analysis of ammonoid sutures. Lethaia, Vol. 43, pp. 266,277. A morphometric method based on eigenshape analysis is proposed for characterizing the morphospace of widely varied ammonoid suture shapes. The analysis requires initially the placement of a suture line in an x,y coordinate system, with the ventral and umbilical extremes located on the x -axis. Series of x- and y -coordinates along the suture line are used as descriptors. The coordinate data are summarized into the two largest principal components using eigenshape analysis. A total of 115 species belonging to six ammonoid orders, spanning from the Devonian to the Cretaceous, was utilized in the present analysis. The analysis, using y -coordinate data, revealed differences in morphological variation in suture shape among taxa within the Mesozoic ammonitids: the Lytoceratina and Ammonitina were characterized by small negative values of the first eigenshape scores, whereas the Phylloceratina (the sister group of the Lytoceratina plus Ammonitina), as well as the Triassic ceratitids and Palaeozoic ammonoids, have a wide range of the first eigenshape scores. The pattern of data obtained from many different ammonoid species, as plotted on eigenshape axes in the morphospace constructed based on y -coordinate data, reveals a plesiomorphic aspect of suture shape in some phylloceratine species with respect to other ammonitids. ,Ammonoids, eigenshape analysis, morphometrics, suture line. [source] Expression of an artificial polypeptide with a repeated tripeptide glutamyl,tryptophanyl,lysine in Saccharomyces cerevisiaeLETTERS IN APPLIED MICROBIOLOGY, Issue 2 2003S.Y. Lee Abstract Aims: Artificial genes, which encode 48 or 64 repeats of a tripeptide, glutamyl,tryptophanyl,lysine have been cloned to the yeast expression vector pAM82 containing the PHO5 promoter and expressed in Saccharomyces cerevisiae AH22. Methods and Results: When the yeast cells harbouring recombinant plasmids pALTG6-2 and pALTG4-4 were derepressed in Burkholder minimal medium (Toh-e, A., Ueda, Y., Kakimoto, S.I. and Oshima, Y. (1973) Journal of Bacteriology113, 727,738) containing low phosphate (0·03 g l,1 KH2PO4 and 1·5 g l,1 KCl), the expression was the highest after 24 h induction and the artificial polypeptides were synthesized to about 10% (pALTG6-2) and 14% (pALTG4-4) of the total cell protein. Conclusions: The artificial polypeptides produced in yeast were made to react with the rabbit antiserum against the polypeptide purified from Escherichia coli and found only in the pellet fraction of cell lysates, indicating the formation of inclusion body. Artificial polypeptide consisting of Glu,Trp,Lys may be useful as partial supplement in food and feeds. Significance and Impact of the Study: The production of single cell enriched with homopolymers of an essential amino acid in yeast might be an important tool of supplementing cereal diets and feed grain rations and could be used as means for improvement of the amino acid profile of single cell protein and production of pharmaceutical peptides. [source] | |||||||||||||||||||||||||||||||