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Evaluation of quadrupole time-of-flight tandem mass spectrometry and ion-trap multiple-stage mass spectrometry for the differentiation of C-glycosidic flavonoid isomers.

LC-MS-MS is becoming a very important tool for the on-line identification of natural products in crude plant extracts. For an efficient use of this technique in the dereplication of natural products, a careful study of the parameters used to generate informative MS-MS spectra is needed. In this paper, the collision-induced dissociation (CID) MS-MS spectra of ubiquitous C-glycosidic flavonoids have been systematically studied using hybrid quadrupole time-of-flight and ion-trap (IT) mass analysers under various CID energy conditions. Efficient differentiation of flavonoid C-glycoside isomers was possible, based on the comparison of CID-MS-MS spectra of particular C-glycoside unit fragments. Striking differences between 6-C and 8-C flavonoid glycosides were especially observed in the product ion spectra of their 0.2X+ fragments ([M+H-120]+). Some guidelines for the on-line characterisation of C-glycosidic flavonoids by LC-MS-MS or LC-multiple-stage MS are given.

Flavonoids↗

The relative importance of intracellular proteolysis and transport on the yield of the periplasmic enzyme penicillin amidase in Escherichia coli*

Intracellular proteolysis is an important mechanism for regulating the level of the periplasmic enzyme penicillin amidase in Escherichia coli. Evidence is presented that the active enzyme is localized in the periplasmic space and maturation of pro-enzyme occurs during transport through the cytoplasmic membrane or rapidly after its entrance in the periplasm. The rate constants of the transport through cytoplasmic membrane and of the intracellular proteolysis were estimated to be 0.01 h and 0.5 h, respectively. This indicates that more than 90% of the synthesized pre-pro-enzyme is lost by intracellular proteolysis occurring in the cytoplasm.

Journal Article↗

Mineralogy and mineral collections in 18th-century France.

During the 18th century, mineralogy constituted an integral part of natural history, sharing the concerns of botany and zoology over collection and classification. In Paris, many people owned private mineral collections, but these have been largely neglected by historians. Here, I examine the place of private collections in the history of mineralogy, arguing that they contributed socially, economically and intellectually to the field in a period before the dominance of the large national collection. I also show how the interests of private collectors diverged from those of the curators of public collections, particularly following the French Revolution.

Catalogs as Topic↗

Peptide mimics of a conformationally constrained protective epitopes of respiratory syncytial virus fusion protein.

AIMS: To identify peptides that mimic (mimotopesi conformational and protective epitopes of RSV fusion protein and to assess their efficacy as immunogens and potential vaccines. MATERIAL AND METHODS: An 8-mer solid-phase (TG resin) library was screened with a neutralising and protective RSV fusion protein specific monoclonal antibodies (Mab-19). After selection of positive beads, reactive sequences were identified by microsequencing and 8-mer peptides were synthesised. Improvement of binding was analysed by amino acid replacement using the SPOTs method. RESULTS: Mabs were not able to bind to the free and soluble peptides, nor did these peptides induce anti-RSV specific antibodies. However, several peptides re-synthesised on a TG resin (to produce de-protected 8-mer peptides linked to the resin) or as SPOTs reacted specifically. Therefore it was critical to be able to reproduce this conformation in order to use these mimotopes as immunogens and potential vaccines. Using C-terminal constrained versions of the mimotopes, strong binding of one of the Mabs to the peptides was demonstrated by surface-plasmon resonance. Immunisation of Balb/c mice with these peptide-mimics produced anti-sera that: (1) reacted specifically with RSV; (2) inhibited the binding of the Mab to the virus; (3) neutralised RSV in vitro with high titres (range: 80-640); and (4) reduce significantly the viral load in the lungs of mice challenged with RSV (P < 0.01). CONCLUSIONS: This report demonstrates for the first time that: (1) a protective epitope of the conserved RSV fusion protein can be mimicked by synthetic peptides; and (2) immunisations with these mimotopes induced specific anti-RSV neutralising antibodies and reduced viral load in vivo. These results represent a novel concept for the development of a vaccine against RSV.

Animals↗

Allergy to asticot maggots. Identification of allergens.

UNLABELLED: There are some written reports of allergic reactions after contact with Asticot larvae in anglers and occupationally exposed workers. Clinical and immunological studies were performed to confirm the allergens involved in the case of a 12-year-old male who developed respiratory symptoms shortly after contact with Asticot maggots used as fish bait. METHODS AND RESULTS: Aqueous extracts of Asticot maggots were taken for in vivo and in vitro tests. The protein concentration as determined by turbidimetric assay was 1.4 mg/ml. Skin prick test with Asticot extract was positive with an immediate response. Ten atopic and ten nonatopic control subjects did not react to the extract. Specific serum IgE antibodies against Asticot were found in the patient's serum (ELISA). Coomasie staining after SDS-PAGE separation of Asticot extract showed three protein bands of 30, 40 and 60 kDa. IgE immunoblot showed one antigenic band of 60 kDa specifically recognized by the patient's IgE. Specific nasal challenge test with Asticot extract using a Rhinospir 164 rhinomanometer showed immediate response (1/10 w/v), with a 200% increase in nasal resistances from baseline. CONCLUSIONS: One main allergen of MW 60 kDa was specifically recognized by our patient's IgE. There are no previous reports in the literature that characterize the allergens involved in Asticot hypersensitivity. Asticot maggots should be taken into account as a possible causative agent of respiratory symptoms due to a type I hypersensitivity mechanism in anglers who are exposed to emanations of these live fish baits.

Allergens↗

Antidepressant effects on kinase gene expression patterns in rat brain.

Multiple kinase pathways determine serotonin transporter (SERT) regulation. We hypothesized a decrease in kinase expression with chronic selective serotonin reuptake inhibitor (SSRI) administration necessary to regulate extracellular serotonin. We studied whole brain kinase mRNA expression on Affymetrix gene chips in rats treated with placebo 3 and 21 days, fluoxetine 3 and 21 days, and citalopram 21 days. Protein kinase C (PKC)-delta, PKC-gamma, stress-activated protein kinase, cAMP-dependent protein kinase beta isoform, Janus protein kinase, and phosphofructokinase M were all down regulated chronically with citalopram and fluoxetine, but not with acute fluoxetine. The results are consistent with homeostasis of SERT function through a decrease in PK expression.

Animals↗