Pyridine nucleotide metabolism in Escherichia coli. II. Niacin starvation

Journal ArticleThe effect of niacin starvation has been studied in a niacin-requiring auxotroph of Escherichia coli. If a culture is totally deprived of niacin, cells continue to divide until the total pyridine nucleotide content has fallen from 1.9 X 10^6 to 1.2 X 10^5 molecules per cell. During starvation, the relative proportion of the pyridine nucleotides changes greatly: the TPN: DPN ratio increases from 0.30 to over 2.0 and nicotinic acid mononucleotide accumulates until it is present at concentrations comparable to DPN

Pyridine nucleotide metabolism in Escherichia coli. I. Exponential growth

Journal ArticleThe pyridine nucleotide pool of Escherichia coli is made up almost exclusively of DPN and TPN. In an exponentially growing glucose culture an average cell contains 1,460,000 molecules of DPN (30% reduced) and 440,000 molecules of TPN (57% reduced). If the total volume of the cell were accessible to pyridine nucleotide, this would mean a total DPN concentration of 1.3 X 10-3 M and a TPN concentration of 3.9 X 10-4 M

Molecular signatures of cell migration in C. elegans Q neuroblasts.

Metazoan cell movement has been studied extensively in vitro, but cell migration in living animals is much less well understood. In this report, we have studied the Caenorhabditis elegans Q neuroblast lineage during larval development, developing live animal imaging methods for following neuroblast migration with single cell resolution. We find that each of the Q descendants migrates at different speeds and for distinct distances. By quantitative green fluorescent protein imaging, we find that Q descendants that migrate faster and longer than their sisters up-regulate protein levels of MIG-2, a Rho family guanosine triphosphatase, and/or down-regulate INA-1, an integrin alpha subunit, during migration. We also show that Q neuroblasts bearing mutations in either MIG-2 or INA-1 migrate at reduced speeds. The migration defect of the mig-2 mutants, but not ina-1, appears to result from a lack of persistent polarization in the direction of cell migration. Thus, MIG-2 and INA-1 function distinctly to control Q neuroblast migration in living C. elegans

Identification of a Putative Crf Splice Variant and Generation of Recombinant Antibodies for the Specific Detection of Aspergillus fumigatus

BACKGROUND: Aspergillus fumigatus is a common airborne fungal pathogen for humans. It frequently causes an invasive aspergillosis (IA) in immunocompromised patients with poor prognosis. Potent antifungal drugs are very expensive and cause serious adverse effects. Their correct application requires an early and specific diagnosis of IA, which is still not properly achievable. This work aims to a specific detection of A. fumigatus by immunofluorescence and the generation of recombinant antibodies for the detection of A. fumigatus by ELISA. RESULTS: The A. fumigatus antigen Crf2 was isolated from a human patient with proven IA. It is a novel variant of a group of surface proteins (Crf1, Asp f9, Asp f16) which belong to the glycosylhydrolase family. Single chain fragment variables (scFvs) were obtained by phage display from a human naive antibody gene library and an immune antibody gene library generated from a macaque immunized with recombinant Crf2. Two different selection strategies were performed and shown to influence the selection of scFvs recognizing the Crf2 antigen in its native conformation. Using these antibodies, Crf2 was localized in growing hyphae of A. fumigatus but not in spores. In addition, the antibodies allowed differentiation between A. fumigatus and related Aspergillus species or Candida albicans by immunofluorescence microscopy. The scFv antibody clones were further characterized for their affinity, the nature of their epitope, their serum stability and their detection limit of Crf2 in human serum. CONCLUSION: Crf2 and the corresponding recombinant antibodies offer a novel approach for the early diagnostics of IA caused by A. fumigatus