CAROTENOID COMPOSITION OF MARINE RED ALGAE1

Photosynthetic organisms possess carotenoids that function either as accessory, photoprotective, or structural pigments. Therefore, the carotenoid profile provides information about certain photoacclimation and photoprotection responses. Carotenoids are also important chemosystematic markers because specific enzymes mediate each step of carotenoid biosynthesis. For red algae, diverse and often contradictory carotenoid compositions have been reported. As a consequence, it is difficult to infer the physiological importance of carotenoids in Rhodophyta. To characterize the relationship between carotenoid composition, rhodophycean phylogeny, and the presence of potentially photoprotective pigments, we analyzed the carotenoid composition of 65 subtropical species from 12 orders and 18 rhodophyte families. Our results showed that red algae do not present a unique carotenoid profile. However, a common profile was observed up to the level of order, with exception of the Ceramiales and the Corallinales. The main difference between profiles is related to the xanthophyll that represents the major carotenoid. In some species lutein is the major carotenoid while in others it is substituted by zeaxanthin or antheraxanthin. The presence of this epoxy carotenoid together with the presence of violaxanthin that are xanthophyll cycle (XC)‐related pigments was found in four of the 12 analyzed orders. The carotenoid pigment profiles are discussed in relation to Rhodophyta phylogeny, and it is suggested that the xanthophyll cycle‐related pigments appeared early in the evolution of eukaryotic phototrophs.     

Die respiratorischen anpassungserscheinungen bei den puppen der simuliiden

Ohne Zusammenfassung     

Evidence for regulation of amelogenin gene expression by 1,25‐dihydroxyvitamin D3 in vivo

The unique hereditary enamel defect clearly related to the disturbance of one enamel matrix protein is X‐linked amelogenesis imperfecta (AI), in which several mutations of amelogenin gene have been identified. The clinical phenotype of many of these subjects shows similarities with enamel defects related to rickets. Therefore, we hypothesized that rachitic dental dysplasia is related to disturbances in the amelogenin pathway. In order to test this hypothesis, combined qualitative and quantitative studies in experimental vitamin D‐deficient (−D) rat model systems were performed. First, Western blot analysis of microdissected enamel matrix (secretion and maturation stages) showed no clear evidence of dysregulation of amelogenin protein processing in −D rats as compared with the controls. Second, the ultrastructural investigation permitted identification of the internal tissular defect of rachitic enamel, the irregular absence of intraprismatic enamel observed in −D animals, suggesting a possible link between prism morphogenesis and vitamin D. In addition, the steady‐state levels of amelogenin mRNAs measured in microdissected dental cells was decreased in −D rats and up‐regulated by an unique injection of 1,25‐dihydroxyvitamin D₃ (1,25(OH)₂D₃). The present study shows evidences that amelogenin expression is regulated by vitamin D. This is the first study of an hormonal regulation of tooth‐specific genes. J. Cell. Biochem. 76:194–205, 1999. © 1999 Wiley‐Liss, Inc.     

Chemoselectively addressable HCan building blocks in peptide synthesis: L-homocanaline derivatives

(S-2-amino-5-(aminooxy)pentanoic acid (L -homocanaline, HCan), a structural analogue of lysine, contains a reactive alkyloxyamine side chain and is therefore considered to react chemoselectively with carbonyl compounds by forming a kinetically stable oxime bond. The chemical synthesis of L -homocanaline starting from protected glutamic acid derivatives is described. Two orthogonally protected homocanaline derivatives were synthesized and their use in standard SPPS procedures was exemplified for the synthesis of a chemoselectively addressable cyclic peptide for use in TASP design. Moreover, the wide range of applications of this unique building block was demonstrated for the chemoselective ligation of an unprotected disaccharide to a HCan containing model peptide resulting in a chimeric glycopeptide structure. © 1998 European Peptide Society and John Wiley & Sons, Ltd.     

Functional differences between TRPC4 splice variants.

Functional characterizations of heterologously expressed TRPC4 have revealed diverse regulatory mechanisms and permeation properties. We aimed to clarify whether these differences result from different species and splice variants used for heterologous expression. Like the murine beta splice variant, rat and human TRPC4beta both formed receptor-regulated cation channels when expressed in HEK293 cells. In contrast, human TRPC4alpha was poorly activated by stimulation of an H(1) histamine receptor. This was not due to reduced expression or plasma membrane targeting, because fluorescent TRPC4alpha fusion proteins were correctly inserted in the plasma membrane. Furthermore, currents through both human TRPC4alpha and TRPC4beta had similar current-voltage relationships and single channel conductances. To analyze the assembly of transient receptor potential channel subunits in functional pore complexes in living cells, a fluorescence resonance energy transfer (FRET) approach was used. TRPC4alpha and TRPC4beta homomultimers exhibited robust FRET signals. Furthermore, coexpressed TRPC4alpha and TRPC4beta subunits formed heteromultimers exhibiting comparable FRET signals. To promote variable heteromultimer assemblies, TRPC4alpha/TRPC4beta were coexpressed at different molar ratios. TRPC4beta was inhibited in the presence of TRPC4alpha with a cooperativity higher than 2, indicating a dominant negative effect of TRPC4alpha subunits in heteromultimeric TRPC4 channel complexes. Finally, C-terminal truncation of human TRPC4alpha fully restored the channel activity. Thus, TRPC4beta subunits form a receptor-dependently regulated homomultimeric channel across various species, whereas TRPC4alpha contains a C-terminal autoinhibitory domain that may require additional regulatory mechanisms.     

The mechanism of assembly and cofactor insertion into Rhodobacter capsulatus xanthine dehydrogenase

Rhodobacter capsulatus xanthine dehydrogenase (XDH) is a molybdo-flavoprotein that is highly homologous to the homodimeric mammalian xanthine oxidoreductase. However, the bacterial enzyme has an (alphabeta)(2) heterotetrameric structure, and the cofactors were identified to be located on two different polypeptides. We have analyzed the mechanism of cofactor insertion and subunit assembly of R. capsulatus XDH, using engineered subunits with appropriate substitutions in the interfaces. In an (alphabeta) heterodimeric XDH containing the XdhA and XdhB subunits, the molybdenum cofactor (Moco) was shown to be absent, indicating that dimerization of the (alphabeta) subunits has to precede Moco insertion. In an (alphabeta)(2) XDH heterotetramer variant, including only one active Moco-center, the active (alphabeta) site of the chimeric enzyme was shown to be fully active, revealing that the two subunits act independent without cooperativity. Amino acid substitutions at two cysteine residues coordinating FeSI of the two [2Fe-2S] clusters of the enzyme demonstrate that an incomplete assembly of FeSI impairs the formation of the XDH (alphabeta)(2) heterotetramer and, thus, insertion of Moco into the enzyme. The results reveal that the insertion of the different redox centers into R. capsulatus XDH takes place sequentially. Dimerization of two (alphabeta) dimers is necessary for insertion of sulfurated Moco into apo-XDH, the last step of XDH maturation.     

Reactions of Airway Epithelial Cells to Birch Pollen Grains Previously Exposed to In Situ Atmospheric Pb Concentrations: A Preliminary Assay of Allergenicity

A growing body of evidence suggests that interactions between pollen grains and environmental pollutants, especially air pollutants, could be of critical importance with regard to the increase in allergic responses observed in the past decades. Using birch pollen grains (BPG), a major allergy source in European countries, and lead (Pb), a highly toxic metal trace element (MTE) present in urban areas, the immune response of human epithelial cells exposed to BPG or to Pb-associated BPG was compared. The cellular response after exposure either to BPG, BPG exposed to 30?mg/L of Pb (BPG-30), or BPG exposed to 60?mg/L of Pb (BPG-60) was evaluated after two time lapses (2 and 6?h) by measuring mRNA levels of four mediators, including two inflammatory (interleukin-8 and interleukin-6) and two allergic (interleukin-5 [IL-5] and interleukin-13) cytokines. After 2?h of exposure, significant upregulation of the IL-5 gene was observed after exposure to BPG-60 in comparison with exposure to BPG and BPG-30 (N _IL-5?=?1.9, Mann?CWhitney test, p?=?0.003). After 6?h of exposure, significant upregulation of the IL-5 gene was observed after exposure to BPG-30 with N _IL-5?=?1.8 and to BPG-60 with N _IL-5?=?2.3 (Mann?CWhitney test, p?=?0.0029) in comparison with exposure to BPG. This first attempt to investigate the influence of pollution by MTE on pollen grain showed a dose?Ctime-dependent increase in IL-5 gene expression after exposure to BPG combined to Pb.