Role of rhizobial lipo-chitin oligosaccharide signal molecules in root nodule organogenesis

The role of oligosaccharide molecules in plant development is discussed. In particular the role of the rhizobial lipo-chitin oligosaccharide (LCO) signal molecules in the development of the root nodule indicates that oligosaccharides play an important role in organogenesis in plants. Recent results of the analyses of structures and of the biosynthesis of the LCO molecules are summarized in this paper. The knowledge and technologies that resulted from these studies will be important tools for further studying the function of LCO signals in the plant and in the search for analogous signal molecules produced by plants.     

Gene isolation through genomic complementation using an indexed library of Chlamydomonas reinhardtii DNA

Hundreds of mutants with defects in a variety of physiologically important functions, such as photosynthesis, respiration, flagellar motility, phototaxis, circadian rhythms and the cell cycle, have been isolated from cultures of Chlamydomonas reinhardtii. In only a few cases have the genes responsible for these mutations been cloned and sequenced. The development of efficient methods for transformation with nuclear genes [7] has allowed the recent demonstration of gene isolation through genomic complementation with a pooled library of C. reinhardtii DNA [9]. To improve the efficiency with which genes complementing a particular mutation can be isolated, we have established an indexed (ordered) cosmid library of 11,280 individual clones contained in the separate wells of 120 microtiter plates. The average insert size is ca. 38 kb. PCR analysis of five sequenced nuclear genes present in the Chlamydomonas library revealed a range from two copies for the ₂ and ₂ tubulin genes to at least seven copies for the agininosuccinate lyase gene. Overall, these five clones were represented an average of >-3.4 times in the library. Thus, the probability that any one particular nuclear gene of < 1000 bp will be found in the library is >-97%, and the probability that a gene of ca. 10 000 bp will be found in the library is ca. 92%. Rapid screening methods with cosmid DNAs pooled from individual microtiter dishes have been applied successfully. Bacteria containing clones of the argininosuccinate lyase gene have been identified through genomic complementation of a Chlamydomonas mutant bearing an inactive arginnosuccinate lyase gene.We are using the nomenclature of indexed library versus ordered library to avoid confusion of this library with a library of ordered contigs.     

Influence of nitrogen,phosphorus and potassium on chemical composition and activity of some enzymes in celery during its growth

The influence of N, P, K supply on the contents of dry mass, chlorophyll (a+b), vitamin C, saccharides and the activities of peroxidase (E.C. 1.11.1.7), catalase (E.C. 1.11.1.6), and acid phosphatase (E.C.3.1.3.2) in leaves of celery plants during their growth stages was investigated. A correlation between the amount of N, P, K and the chemical composition and catalytic activities of enzymes was found.     

Genetic and Physical Mapping of a Gene Encoding a Methyl CpG Binding Protein, Mecp2, to the Mouse X Chromosome

The methyl CpG binding proteins (MeCP1 and MeCP2) are a class of proteins that bind to templates containing symmetrically methylated CpGs. Using an interspecific backcross segregating a number of X-linked markers, we have localized the Mecp2 gene in mouse to the X chromosome close to the microsatellite marker DXMit1. Detailed physical mapping utilizing an available YAC contig encompassing the DXMit1 locus has localized the Mecp2 gene to a 40-kb region between the L1cam and the Rsvp loci, indicating the probable position of a homologue on the human X chromosome.     

Structure determination of the intact major sialylated oligosaccharide chains of recombinant human erythropoietin expressed in Chinese hamster ovary cells

Recombinant human erythropoietin (rHuEPO) is used abundantlyin the clinic to stimulate red blood cell growth in anaemicpatients. The efficacy of the drug depends strongly on the extentof sialylation of its carbohydrate moiety. Prompted by conflictingliterature reports on the issue, we reinvestigated the structuresof the intact sialylated carbohydrate chains of rHuEPO expressedin Chinese hamster ovary (CHO) cells. The asparagine-linkedoligosaccharides were released from rHuEPO with N-glycanaseand fractionated by anion-exchange chromatography. The O-linkedoligosaccharides were released under alkaline borohydride conditions.The primary structures of the major sialylated N- and O-typeoligosaccharides were identified by 500-MHz ¹H-NMR spectroscopy,supported by data from composition analysis, methylation analysis,low- and high-pH anion-exchange chromatography, and fast atombombardment-mass spectrometry. The mod abundant N-linked oligosaccharidesin CHO cell-derived rHuEPO were found to be di-antennary, 2,4-branchedtri-antennary, 2,6-branched tri-antennary and tetra-antennarychains (in the ratio of 7:6:5:82), with the latter containingbetween zero and three repeating N-acetyllactosamine units,in well-defined branches. The major (>95%) di-, tri- andtetra-antennary structures are fully sialylated, i.e. they havetwo, three and four sialic acid residues, respectively, Linkedexclusively (23) to galactose residues. The majority (>95%)of N-Linked structures contain (16)-linked fucose at the proximalGlcNAc residue. The O-type mono- and disialyl oligosaccharideswere characterized as a linear tri- and a branched tetra-saccharide,respectively. erythropoietin FAB-MS ¹H-NMR recombinant glycoprotein sialic acid     

Gemfibrozil treatment is associated with elevated adrenal androgen, androstanediol glucuronide and cortisol levels in dyslipidemic men

We have investigated the role of steroid hormones as coronary risk factors in Helsinki Heart Study population of dyslipidemic middle-aged men. We compare here the effects of gemfibrozil and placebo on the serum levels of dehydroepiandrosterone (DHEA), its sulfate (DHEAS), their metabolite androstanediol glucuronide (3-AdiolG), androstenedione, cortisol, testosterone, and sex-hormone binding globulin (SHBG) in non-smokers. We also examined the associations between steroid and lipoprotein levels in both treatment groups. Compared with placebo gemfibrozil treatment was associated with significant elevations of the mean levels of DHEA 10.2 vs 8.0 nmol/1; P<0.005, of DHEAS 8.0 vs 5.8 μmol/1; P<0.001, of 3AdiolG 18.3 vs 8.4 nmol/1; P<0.001, of androstenedione 5.7 vs 5.1 nmol/1; P<0.02, and of cortisol 426 vs 358 nmol/1; P<0.001. The mean SHBG levels decreased from 46.4 to 41.7 nmol/1; P=0.03 with gemfibrozil treatment. No difference was found in testosterone levels 17.7 vs 18.8 nmol/1; P=0.11, or the ratio of testosterone/SHBG 0.45 vs 0.43; P=0.23. Positive correlations were found between high density lipoprotein-cholesterol and DHEAS (r=0.267; P<0.01) and DHEA (r=0.282; P<0.01) levels and negative correlations between low density lipoprotein-cholesterol and 3-AdiolG (r=−0.400; P<0.001) and cortisol (r=−0.281; P<0.01) levels in the gemfibozil group. Our results indicate that gemfibrozil treatment increases the production and turnover of adrenal androgens and cortisol, and suggest that activation of the adrenocorticol function and increased metabolism of androgens are related to the improved lipoprotein pattern during gemfibrozil treatment.