Phyllactinia fraxinicola,another Asian fungal pathogen on Fraxinus excelsior (common ash) introduced to Europe?

Since the mid-nineties Phyllactinia fraxini has become frequent in Germany. This species has hitherto been characterized by having straight conidiophore foot cells. However, we found that recent collections from Germany have conidiophores with sinuated and twisted foot cells. So far sinuated foot cells were only known from the related P. fraxinicola, another species with Eastern Asian origin. We thus hypothesized that recent collections from Germany belong to P. fraxinicola which might have been introduced to Europe. Using morphological and molecular rDNA data we found that no introduction took place and that there is only P. fraxini in Germany.     

Model identification of the biochemical oxidation process

A fairly general model of the biochemical oxidation, which takes into account the activity of microorganisms, is presented. Parameters of the model have been determined by fitting the model to available experimental data through the use of a straightforward gradient technique.     

Erysiphe azerbaijanica and E. linderae: Two new powdery mildew species (Erysiphales) belonging to the Microsphaera lineage of Erysiphe

Two new species, Erysiphe azerbaijanica on Castanea sativa and E. linderae on Lindera praecox, both belonging to the Microsphaera lineage of the genus Erysiphe are described based on morphological and molecular data. Erysiphe azerbaijanica is distinguished from other Erysiphe species occurring on Castanea spp. by its cylindrical conidia with a length/width ratio of 2–3.6, longer conidiophore, and foot-cells. Molecular analyses indicated that this species forms a clade of its own, supporting the morphological observations. Molecular phylogenetic analyses showed that E. blasti s. lat. is divided into two genetically differentiated groups associated with different host species. Based on the sequence differences in the 28S rRNA gene and ITS region, connected with differences in the number and length of appendages, the fungus on L. praecox is described as a new species, E. linderae.     

Fibrinogen Nagoya, a replacement of glutamine-329 by arginine in the gamma-chain that impairs the polymerization of fibrin monomer

Structural studies on a hereditary abnormal fibrinogen, fibrinogen Nagoya (Takamatsu, J., Ogata, K., Kamiya, T., Koie, K., Takagi, T., & Iwanaga, S. (1979) Thromb. Haemost. 42, 78), were performed to identify the abnormality responsible for the impaired polymerization of fibrin monomer. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, fibrinogen Nagoya showed the presence of an extra protein band with an apparent molecular weight of 49,500 in addition to the normal three subunit chains. Amino acid sequence analysis of a peptide isolated from a lysyl endopeptidase digest of one of the CNBr fragments derived from fibrinogen Nagoya indicated that Gln-329 in the gamma-chain had been replaced by Arg. This substitution can be explained by a single nucleotide change in the codon for Gln-329 (CAG----CGG). We conclude that Gln-329 in the gamma-chain is indispensable for the normal polymerization of fibrin monomer.     

Leukotriene A synthase activity of purified mouse skin arachidonate 8-lipoxygenase expressed in Escherichia coli

Mouse skin 8-lipoxygenase was expressed in COS-7 cells by transient transfection of its cDNA in pEF-BOS carrying an elongation factor-1α promoter. When crude extract of the transfected COS-7 cells was incubated with arachidonic acid, 8-hydroxy-5,9,11,14-eicosatetraenoic acid was produced as assessed by reverse- and straight-phase high performance liquid chromatographies. The recombinant enzyme also reacted on α-linolenic and docosahexaenoic acids at almost the same rate as that with arachidonic acid. Eicosapentaenoic and γ-linolenic acids were also oxygenated at 43% and 56% reaction rates of arachidonic acid, respectively. In contrast, linoleic acid was a poor substrate for this enzyme. The 8-lipoxygenase reaction with these fatty acids proceeded almost linearly for 40 min. The 8-lipoxygenase was also expressed in an Escherichia coli system using pQE-32 carrying six histidine residues at N-terminal of the enzyme. The expressed enzyme was purified over 380-fold giving a specific activity of approximately 0.2 μmol/45 min per mg protein by nickel–nitrilotriacetate affinity chromatography. The enzymatic properties of the purified 8-lipoxygenase were essentially the same as those of the enzyme expressed in COS-7 cells. When the purified 8-lipoxygenase was incubated with 5-hydroperoxy-6,8,11,14-eicosatetraenoic acid, two epimers of 6-trans-leukotriene B₄, degradation products of unstable leukotriene A₄, were observed upon high performance liquid chromatography. Thus, the 8-lipoxygenase catalyzed synthesis of leukotriene A₄ from 5-hydroperoxy fatty acid. Reaction rate of the leukotriene A synthase was approximately 7% of arachidonate 8-lipoxygenation. In contrast to the linear time course of 8-lipoxygenase reaction with arachidonic acid, leukotriene A synthase activity leveled off within 10 min, indicating suicide inactivation.     

Expression and purification of biologically active human OSF-1 in Escherichia coli.

OSF-1 (also known as pleiotrophin, HB-GAM, HBGF-8 or HBNF) is a heparin-binding, neurotrophic protein. Its tissue-specific expression in rats is developmentally regulated and the protein is highly conserved between species. The protein is believed to be involved in neuronal development. Previous experiments in our laboratory showed that OSF-1 is primarily expressed in brain and bone. The biological function of OSF-1 in bone is unknown. In order to overcome the limited availability of the native protein, we now report on the high-level expression of human OSF-1 in Escherichia coli. The protein is present in the form of inclusion bodies, which were isolated and solubilized. The partially purified protein was refolded and further purified employing heparin sepharose chromatography. N-terminal sequence determination revealed the same amino acid sequence as the natural mature protein. The isolated backfolded recombinant human OSF-1 did promote neurites outgrowth in primary cultures of cortical neurons.     

Transcriptional regulator SpxA1a controls the resistance of the honey bee pathogen Melissococcus plutonius to the antimicrobial activity of royal jelly

Royal jelly (RJ), a brood food of honey bees, has strong antimicrobial activity. Melissococcus plutonius, the causative agent of European foulbrood of honey bees, exhibits resistance to this antimicrobial activity and infects larvae orally. Among three genetically distinct groups (CC3, CC12 and CC13) of M. plutonius, CC3 strains exhibit the strongest RJ resistance. In this study, to identify genes involved in RJ resistance, we generated an RJ-susceptible derivative from a highly RJ-resistant CC3 strain by UV mutagenesis. Genome sequence analysis of the derivative revealed the presence of a frameshift mutation in the putative regulator gene spxA1a. The deletion of spxA1a from a CC3 strain resulted in increased susceptibility to RJ and its antimicrobial component 10-hydroxy-2-decenoic acid. Moreover, the mutant became susceptible to low-pH and oxidative stress, which may be encountered in brood foods. Differentially expressed gene analysis using wild-type and spxA1a mutants revealed that 45 protein-coding genes were commonly upregulated in spxA1a-positive strains. Many upregulated genes were located in a prophage region, and some highly upregulated genes were annotated as universal/general stress proteins, oxidoreductase/reductase, chaperons and superoxide dismutase. These results suggest that SpxA1a is a key regulator to control the tolerance status of M. plutonius against stress in honey bee colonies.     

Structural characteristics of the ceramides of neutral glycosphingolipids in the human female genital tract--their menstrual cycle-associated change in the cervical epithelium and uterine endometrium, and their dissociation in the mucosa of the fallopian tube with the menstrual cycle.

In human cervical epithelium, uterine endometrium, and mucosa of the fallopian tubes, neutral glycosphingolipids were exclusively represented by the globo-series glycosphingolipids, such as CMH, LacCer, Gb3Cer and Gb4Cer, but the molecular species of their ceramide moieties were characteristically altered in the cervical epithelium and uterine endometrium during the menstrual cycle. Individual neutral glycosphingolipids in the cervical epithelium and the uterine endometrium at the follicular phase gave two bands on TLC, whereas those at the luteal phase displayed three bands, the third being the lower migrating one. Neutral glycosphingolipids migrating to the same positions as these lower-migrating bands were constantly detected in the mucosa of the fallopian tubes, independent of the menstrual cycle. The lower-migrating bands for the cervical epithelium and the uterine endometrium at the luteal phase were due to molecules mainly constructed of phytosphingosine with alpha-hydroxy fatty acids having chain lengths of 18-24 and 4-sphingenine with alpha-hydroxy fatty acids having chain lengths of 16-22, whereas those in the mucosa of the fallopian tubes were exclusively N-alpha-hydroxypalmitoyl 4-sphingenine.