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See detailIsolation of a glycoprotein E-deleted bovine herpesvirus type 1 strain in the field
Dispas, M.; Schynts, F.; Lemaire, Mylène et al

in Veterinary Record : Journal of the British Veterinary Association (2003), 153(7), 209-212

During a field trial to evaluate the efficacy of repeated vaccinations with bovine herpesvirus type 1 (BHV-1) marker vaccines, a glycoprotein E (gE)-negative BHV-1 strain was isolated from the nasal ... [more ▼]

During a field trial to evaluate the efficacy of repeated vaccinations with bovine herpesvirus type 1 (BHV-1) marker vaccines, a glycoprotein E (gE)-negative BHV-1 strain was isolated from the nasal secretions of two cows, eight months after vaccination with a gE-negative live-attenuated vaccine, initially given intranasally, then intramuscularly. The strain isolated was characterised using immunofluorescence, restriction analysis and PCR. All the techniques used identified the isolated virus as a gE-negative BHV-1 phenotypically and genotypically identical to the Za strain used as a control. [less ▲]

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See detailIsolation of a Microsporum canis gene family encoding three subtilisin-like proteases expressed in vivo
Descamps, Frédéric; Brouta, Frédéric; Monod, Michel et al

in Journal of Investigative Dermatology (2002), 119(4), 830-835

Microsporum canis is the main agent of dermatophytosis in dogs and cats and is responsible for frequent zoonosis. The pathogenesis of the disease remains largely unknown, however. Among potential fungal ... [more ▼]

Microsporum canis is the main agent of dermatophytosis in dogs and cats and is responsible for frequent zoonosis. The pathogenesis of the disease remains largely unknown, however. Among potential fungal virulence factors are secreted keratinolytic proteases, whose molecular characterization would be an important step towards the understanding of dermatophytic infection pathogenesis. M. canis secretes a 31.5 kDa keratinolytic subtilisin-like protease as the major component in a culture medium containing cat keratin as the sole nitrogen source. Using a probe corresponding to a gene's internal fragment, which was obtained by polymerase chain reaction, the entire gene encoding this protease named SUB3 was cloned from a M. canis lambdaEMBL3 genomic library. Two closely related genes, termed SUB1 and SUB2, were also cloned from the library using as a probe the gene coding for Aspergillus fumigatus 33 kDa alkaline protease (ALP). Deduced amino acid sequence analysis revealed that SUB1, SUB2, and SUB3 are secreted proteases and show large regions of identity between themselves and with subtilisin-like proteases of other filamentous fungi. Interestingly, mRNA of SUB1, SUB2, and SUBS were detected by reverse transcriptase nested-polymerase chain reaction from hair of experimentally infected guinea pigs. These results show that SUB1, SUB2, and SUB3 encode a family of subtilisin-like proteases and strongly suggest that these proteases are produced by M. canis during the invasion of keratinized structures. This is the first report describing the isolation of a gene family encoding potential virulence-related factors in dermatophytes. [less ▲]

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See detailIsolation of an amylolytic chrysophyte, Poterioochromonas sp. from the digestive tract of the termite R. santonensis
Tarayre, Cédric ULg; Bauwens, Julien ULg; Brasseur, Catherine ULg et al

in Biotechnologie, Agronomie, Société et Environnement = Biotechnology, Agronomy, Society and Environment [=BASE] (2014), 18(1),

The aim of this work was the isolation and cultivation of amylolytic protists living in the digestive tract of the termite Reticulitermes santonensis (Feytaud). A chrysophyte identified as ... [more ▼]

The aim of this work was the isolation and cultivation of amylolytic protists living in the digestive tract of the termite Reticulitermes santonensis (Feytaud). A chrysophyte identified as Poterioochromonas sp. was isolated in a special medium containing rice grains as a source of carbon and nitrogen. Then, the protist was grown in a medium containing starch as a carbon source, tryptone, and a phosphate buffer at different pH values (5, 6 and 7). Yeast extract was added or not. Ciprofloxacin was used to avoid the bacterial development. Other antibiotics were also tested but showed an inhibitive effect on the growth of Poterioochromonas sp. Yeast extract allowed reaching 1.9 (pH 5), 2.3 (pH 6) and 2.2 (pH 7) times higher final cell concentrations, and 2.8 (pH 5), 2.8 (pH 6) and 2.2 (pH 7) times higher biomass yields. The starch concentration did not decrease in the medium until 3 and 4 days of culture, with and without yeast extract, respectively. Eight days of culture were necessary for hydrolyzing the starch completely, with and without yeast extract. Maltose and maltotriose were detected in the culture media and were hydrolyzed progressively. Maximal maltose concentrations were 0.68, 0.66 and 0.51 g.l-1 in the medium containing yeast extract. Maltotriose concentrations were only 0.17, 0.14 and 0.12 g.l-1. Other glucose oligomers were also detected but in lower quantities. It was determined that the protist developed a weak amylase activity, particularly at a weakly acidic pH (5-6). Such a pH also allowed a better growth of the protist. A maximal amylase activity of 112 nkat.l-1 was measured with yeast extract at pH 5. No other enzymatic activity (protease, cellulase or xylanase) was detected except amylase. The degradation products of starch which were obtained by enzymatic hydrolysis allow the identification of α-amylase, amyloglucosidase and possibly β-amylase activities. [less ▲]

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See detailIsolation of an n-alkylated benzylamine derivative from Pseudomonas putida BTP1 as elicitor of induced systemic resistance in bean
Ongena, MARC ULg; Jourdan, Emmanuel ULg; Schafer, M. et al

in Molecular Plant-Microbe Interactions (2005), 18(6), 562-569

Root treatment of Phaseolus vulgaris with the nonpathogenic Pseudomonas putida BTP1 led to significant reduction of the disease caused by the pathogen Botrytis cinerea on leaves. The molecular determinant ... [more ▼]

Root treatment of Phaseolus vulgaris with the nonpathogenic Pseudomonas putida BTP1 led to significant reduction of the disease caused by the pathogen Botrytis cinerea on leaves. The molecular determinant of P putida BTP1 mainly responsible for the induced systemic resistance (ISR) was isolated from cell-free culture fluid after growth of the strain in the iron-poor casamino acid medium. Mass spectrometry analyses performed on both the bacterial product and synthetic analogues revealed a polyalkylated benzylamine structure, with the quaternary ammonium substituted by methyl, ethyl, and C-13 aliphatic groups responsible for the relative hydrophobicity of the molecule. The specific involvement of the N-alkylated benzylamine derivative (NABD) in ISR elicitation was first evidenced by testing the purified compound that mimicked the protective effect afforded by crude supernatant samples. The evidence was supported by the loss of elicitor activity of mutants impaired in NABD biosynthesis. Our experiments also showed that other iron-regulated metabolites secreted by the strain are not involved in ISR stimulation. Thus, these results indicate a wider variety of Pseudomonas determinants for ISR than reported to date. [less ▲]

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See detailIsolation of bacteriocin-producing lactic acid bacteria from fermented fish product in Senegal.
Dortu, C.; Roblain, D.; Totte, A. et al

Poster (2003, May)

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See detailIsolation of bacteriocin-producing lactic acid bacteria from fermented fish product in Senegal.
Dortu, C.; Roblain, D.; Totte, A. et al

Poster (2003, August)

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See detailIsolation of bioactive peptides from tryptone that modulate lipase production in Yarrowia lipolytica
Turki, Souassen; Ben Kraeim, Ines; Weekers, F. et al

in Bioresource Technology (2009), 100

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See detailIsolation of Bovine Follicular Dendritic Cells Allows the Demonstration of a Particular Cellular Prion Protein
Thielen, Caroline ULg; Mélot, France ULg; Jolois, Olivier ULg et al

in Cell & Tissue Research (2001), 306(1), 49-55

As interaction of cellular prion protein (PrPc) and the infectious agent (PrPres) appears to be a crucial pathogenic step promoted by homology, variation in PrPc isoforms on bovine immune cells may ... [more ▼]

As interaction of cellular prion protein (PrPc) and the infectious agent (PrPres) appears to be a crucial pathogenic step promoted by homology, variation in PrPc isoforms on bovine immune cells may explain the absence of infectivity in most bovine lymph organs. In this study, we examined PrPc expression in bovine lymph organs (tonsils and lymph nodes) and on isolated follicular dendritic cells (FDCs). We used a panel of different monoclonal antibodies (MoAbs) raised against different epitopes of prion protein. Two MoAbs recognise amino acids 79-92 (SAF 34 and SAF 32 Mo-Abs); the 6H4 antibody reacts with a specific peptide comprising the 144-152 amino acids, and the 12F10 MoAb recognises the sequence 142-160. After immunolabelling of frozen sections of lymph organs with 6H4 or 12F10 MoAbs, we detected cellular prion protein in germinal centres. However, using the SAF 34 or SAF 32 antibodies, PrPc was revealed outside the lymphoid tissues. No PrPc was observed in the germinal centres. Therefore, we adapted the method of FDC isolation, making it suitable for the study of PrPc expression on their surface. Using electron microscopy, the presence of PrPc on the surface of FDCs was demonstrated only with 6H4 MoAb. These results suggest that bovine follicular dendritic cells express a particular form of prion protein. Either the N-terminal part of PrPc is cleaved or the accessibility of the specific epitope (79-92) of SAF 34 MoAb is abolished by interaction with other molecules. This particular isoform of PrPc on bovine FDCs might be related to the apparent absence of infectivity in lymph organs in cattle affected by bovine spongiform encephalopathy. [less ▲]

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See detailIsolation of caprine herpesvirus type 1 in Spain
Keuser, V.; Espejo-Serrano, J.; Schynts, F. et al

in Veterinary Record : Journal of the British Veterinary Association (2004), 154(13), 395-399

Two strains of caprine herpesvirus type 1 (CpHV-1) were isolated after the experimental reactivation of two seropositive goats in Spain. Viral DNA from these isolates was compared with DNA from bovine ... [more ▼]

Two strains of caprine herpesvirus type 1 (CpHV-1) were isolated after the experimental reactivation of two seropositive goats in Spain. Viral DNA from these isolates was compared with DNA from bovine herpesvirus type 1 and CpHV-1 reference strains by restriction endonuclease analysis. The two Spanish isolates were closely related but could easily be distinguished from each other and from the reference strains. [less ▲]

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See detailIsolation of Clostridium perfringens from three neonatal calves with haemorrhagic abomasitis.
Manteca, C.; Jauniaux, Thierry ULg; Daube, Georges ULg et al

in Revue de Médecine Vétérinaire (2001), 152

Braxy-like disease with sudden death and acute haemorhagic abomasitis was diagnosed in three Belgian Blue calves : one two-day-old and one one month-old calves, in good condition with no clinical signs ... [more ▼]

Braxy-like disease with sudden death and acute haemorhagic abomasitis was diagnosed in three Belgian Blue calves : one two-day-old and one one month-old calves, in good condition with no clinical signs noted a few hours prior to death, and another two day-old calf, which had shown problems of abomasal dilatation and regurgitation prior to death. Histologically, the abomasal wall were oedematous and emphysematous. A pure and abundant growth of Clostridium perfringens was obtained in anaerobic conditions from the abomasal wall of the three Belgian Blue calves. No bacterial growth was obtained in aerobic conditions. The calf with digestive disorders was also positive for BVD virus by immunofluorescence in the abomasal wall and in the spleen. [less ▲]

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See detailIsolation of DD carboxypeptidase from Streptomyces albus G culture filtrates
Ghuysen, Jean-Marie ULg; Leyh-Bouille, Mélina; Bonaly, Roger et al

in Biochemistry (1970), 9(15), 2955-2961

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See detailIsolation of five new monosomic alien addition lines of Gossypium australe F. Muell in G. hirsutum L. by SSR and GISH analyses
Sarr, D.; Lacape, J.-M.; Rodier-Goud, M. et al

in Plant Breeding (2010), 130

Gossypium australe F. Muell (2n + 2x = 26) is a wild perennial species possessing agronomic useful traits that would be interesting to introgress into G. hirsutum L. (2n = 4x = 52), the main cultivated ... [more ▼]

Gossypium australe F. Muell (2n + 2x = 26) is a wild perennial species possessing agronomic useful traits that would be interesting to introgress into G. hirsutum L. (2n = 4x = 52), the main cultivated cotton species. To isolate monosomic alien addition lines (MAALs) of G. australe in G. hirsutum, the [2(G. hirsutum X G. australe) X G. hirsutum] pentaploid (2n = 5x = 65) was backcrossed as male parent to G. hirsutum. Analysis of 42 BC1 plants and seven alien addition lines, already available, with 150 SSR markers developed from G. hirsutum revealed a cross-species amplification rate of 100% and a polymorphism rate of 56%. Eighty polymorphic SSR markers generated 87 G. australe-specific loci that have been assigned by a hierarchical cluster analysis to 13 linkage groups corresponding to the 13 chromosomes of G. australe. Analysis by SSR markers and genomic in situ hybridization of the self-progeny of disomic alien addition lines, backcross progeny of the pentaploid, allowed the isolation of five new MAALs. [less ▲]

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See detailIsolation of follicular dendritic cells from human tonsils and adenoids. I. Procedure and morphological characterization.
Lilet, C.; Radoux, D.; Heinen, Ernst ULg et al

in Journal of Immunological Methods (1984), 66(2), 235-44

Follicular dendritic cells have been isolated from human tonsils and adenoids and characterized at the ultrastructural level. Follicles were dissected and digested with different hydrolytic enzymes. The ... [more ▼]

Follicular dendritic cells have been isolated from human tonsils and adenoids and characterized at the ultrastructural level. Follicles were dissected and digested with different hydrolytic enzymes. The cells were separated by sedimentation at unit gravity. By this procedure we obtained follicular dendritic cells enveloping lymphocytes with their cytoplasmic extensions in a way analogous to that described for isolated thymic nurse cells. The ultrastructural features of isolated follicular dendritic cells are similar to those observed in situ. Prolonged enzymatic action caused loss of the enveloped lymphocytes. [less ▲]

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See detailIsolation of follicular dendritic cells from human tonsils and adenoids. II. Immunocytochemical characterization.
Heinen, Ernst ULg; Lilet, Chantal ULg; Mason, D. Y. et al

in European Journal of Immunology (1984), 14(3), 267-73

Follicular dendritic cells (FDC) are specialized cells found only within lymphoid follicles. They bind immune complexes and play a role in the presentation of antigen to follicular B cells and in the ... [more ▼]

Follicular dendritic cells (FDC) are specialized cells found only within lymphoid follicles. They bind immune complexes and play a role in the presentation of antigen to follicular B cells and in the generation of B cell memory. In the present report the isolation of FDC from human tonsils and adenoids is described. These isolated cells have an unusual spherical arrangement and enclose lymphocytes within extensions of their membranes. Their ultrastructural features are similar to those observed in situ. The reactivity of isolated FDC with a number of monoclonal antibodies was analyzed by immunofluorescence and by immunostaining (at the electron microscopic level) with colloidal gold. In keeping with the results of previous investigations on tissue sections IgM, IgG and IgA (but not IgD) can be detected on the surface of isolated FDC, as can C3b receptors and the FDC-associated antigen detected by monoclonal antibody R4/23. The immunoglobulins associated with FDC are mostly embedded in an electron-dense material. The majority of the lymphoid cells enclosed within the membrane extensions of FDC are of B cell type. These results suggest that isolated FDC may be suitable for further in vitro investigation of their role in the humoral immune response. [less ▲]

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See detailIsolation of follicular dendritic cells from human tonsils and adenoids. III. Analysis of their Fc receptors.
Heinen, Ernst ULg; Radoux, D.; Kinet-Denoel, C. et al

in Immunology (1985), 54(4), 777-84

Follicular dendritic cells (FDC), isolated from human tonsils or adenoids, were tested for their capacity to retain monomeric, aggregated or antigen-bound human antibodies in the absence of serum. FDC ... [more ▼]

Follicular dendritic cells (FDC), isolated from human tonsils or adenoids, were tested for their capacity to retain monomeric, aggregated or antigen-bound human antibodies in the absence of serum. FDC retain fluorescein-labelled heat-aggregated human immunoglobulins, but not monomeric ones nor fluorescein-labelled F(ab')2 in monomeric or aggregated form. Ultrastructural observations showed that colloidal gold-labelled monomeric, or antigen-bound, antibodies directed against tetanus toxoid are retained by dendrites and membrane infoldings of FDC but are never located in cytoplasmic vesicles. This retention was inhibited by incubating FDC with unlabelled aggregated or antigen-bound antibodies. When gold-labelled anti-tetanus toxoid antibodies were incubated in the presence of protein-A before the contact with FDC, a strong reduction of their retention occurred. This further suggested the presence of Fc receptors on isolated tonsillar FDC. Endocytosis was not observed in isolated FDC, even after prolonged incubation in presence of labelled immune complexes: their Fc receptors are, thus, not related to a phagocytic activity as they are in macrophages. Simultaneous ultrastructural labelling of Fc and C3b receptors with colloidal gold particles of different sizes did not reveal any clear relations between these two receptors on the surface of FDC. [less ▲]

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See detailIsolation of follicular dendritic cells from human tonsils and adenoids. In vitro culture.
Cormann, N.; Heinen, Ernst ULg; Kinet-Denoel, C. et al

in Advances in Experimental Medicine and Biology (1988), 237

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See detailIsolation of follicular dendritic cells from human tonsils and adenoids. V. Effect on lymphocyte proliferation and differentiation.
Cormann, N.; Lesage, F.; Heinen, Ernst ULg et al

in Immunology Letters (1986), 14(1), 29-35

Follicular dendritic cells (FDC) are located only in lymph follicles and are characterized by their capacity to retain high amounts of immune complexes on their plasma membranes. As their functions in ... [more ▼]

Follicular dendritic cells (FDC) are located only in lymph follicles and are characterized by their capacity to retain high amounts of immune complexes on their plasma membranes. As their functions in germinal centres are unknown, we isolated them from human tonsils and cultured them with autologous lymphoid cells. Cultures of lymphoid cells alone or with added macrophages were used as controls. Lymphoid cells incorporated tritiated thymidine only when FDC and lectins were added; this could be shown after several periods of time. However, the Ig secretion by lymphoid cell populations was inhibited by FDC after several days in vitro. In contrast, the supernatants of lymphocytes cultured alone or with macrophages only for the same periods of time contained increasing amounts of immunoglobulins. This inhibitory effect of FDC on immunoglobulin production was observed for all considered isotypes. Our data suggest that FDC stimulate lymphoid cell proliferation but reduce B-cell differentiation. This is the first accessory cell activity definitely shown for FDC in cultures. [less ▲]

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See detailIsolation of follicular dendritic cells from human tonsils and adenoids. VI. Analysis of prostaglandin secretion.
Heinen, Ernst ULg; Cormann, N.; Braun, M. et al

in Annales de l'Institut Pasteur. Immunologie (1986), 137D(3), 369-82

Follicular dendritic cells (FDC) are able to fix high amounts of immune complexes by C3b or Fc receptors without endocytosis and for long periods of time. In order to determine the function of this ... [more ▼]

Follicular dendritic cells (FDC) are able to fix high amounts of immune complexes by C3b or Fc receptors without endocytosis and for long periods of time. In order to determine the function of this retention, we analysed the secretion of prostaglandin E2 (PGE2) by FDC in vitro; indeed, it is well-known that immune complex fixation on cells may induce PGE2 production. FDC were isolated by enzymic digestion of lymph follicles dissected under the biomicroscope from human tonsils or adenoids. Isolated FDC appeared as spherical clusters where they enveloped lymphoid cells with their cytoplasmic extensions. Tests were performed in synthetic culture media or in media supplemented with foetal calf serum. PGE2 production in FDC suspensions was compared to that of lymphocyte or macrophage-enriched populations prepared from the same human tonsils. In all experimental conditions, FDC and macrophage-enriched cell populations produced high levels of PGE2, inversely to lymphoid cell populations. This secretion was inhibited by indomethacin. At the ultrastructural level, we also showed that 3H-arachidonic acid was metabolized in cell membranes of all three cell types. The PGE2 produced in the culture media, according to our experimental conditions, do not influence cell proliferation, as assessed by 3H-thymidine incorporation tests on phytohaemagglutinin-stimulated lymphocytes. [less ▲]

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