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See detailIsolation and characterisation of new spore-forming lactic acid bacteria with prospects of use in food fermentations and probiotic preparations
Bayane, Ali; Diawara, B.; Dubois Dauphin, Robin ULg et al

in African Journal of Microbiology Research [=AJMR] (2010), 4(11), 1016-1025

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See detailIsolation and characterisation of seven alien monosomic addition lines of Gossypium australe F. Muell. on G. hirsutum L.
Ahoton, L.; Lacape, J. M.; D'Hont, A. et al

in World Cotton Research Conference - 3. Cotton production for the new Millenium (2004)

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See detailIsolation and characterization of bovine parainfluenza virus type 3 from water buffaloes (Bubalus bulalis) in Argentina
Maidana, S.; Lomonaco, M.; Odeon, A. et al

in BMC Veterinary Research (2012), 8

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See detailIsolation and characterization of eight pregnancy-associated glycoproteins present at high levels in the ovine placenta between day 60 and day 100 of gestation
El Amiri, B.; Remy, Benoit; Melo de Sousa, Noelita ULg et al

in Reproduction Nutrition Development (2004), 44(3, May-Jun), 169-181

Pregnancy-associated glycoproteins (PAG), structurally related to aspartic proteinases, are expressed in the outer epithelial cell layer (chorion/trophectoderm) of the ungulate placenta. The aim of the ... [more ▼]

Pregnancy-associated glycoproteins (PAG), structurally related to aspartic proteinases, are expressed in the outer epithelial cell layer (chorion/trophectoderm) of the ungulate placenta. The aim of the present study was to isolate as many PAG molecules as possible from placentae collected between day 60 and day 100 of gestation and to characterize their amino-terminal amino-acid sequences. Three heterologous radioimmunoassays were used to monitor PAG immunoreactivity throughout the isolation procedures. Sequential use of DEAE-cellulose, gel filtration, and CM ceramic chromatographies led to the isolation of several fractions rich in PAG immunoreactivity. The fractions with a large amount of proteins were also purified by chromatofocusing. The analysis of immunoreactive fractions by SDS-PAGE, Western blotting and two-dimensional electrophoresis followed by amino-terminal microsequencing on PVDF membranes allowed to identify eight different ovPAG with apparent molecular masses ranging from 55 to 66 kDa and isoelectric points from 4.0 to 6.8. The N-terminal sequences were determined and their comparison to those previously identified revealed that four of them are identical to those encoded by previously known cDNA, while the additional four sequences appear to be novel since they have not yet been described. [less ▲]

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See detailThe isolation and characterization of equine Osteocalcin
Carstanjen, B; Wattiez, R; Amory, Hélène ULg et al

in Proceedings of the 1st meeting of the International Bone and Mineral Society (IBMS) (2001)

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See detailIsolation and characterization of mutants deficient in four steps of the phylloquinone biosynthesis pathway in Chlamydomonas reinhardtii.
Emonds-Alt, Barbara ULg; Remacle, Claire ULg; Cardol, Pierre ULg

Poster (2016, April 26)

In photosystem I (PSI), phylloquinone participates to electron transfer as secondary electron acceptor (A1). The phylloquinone biosynthesis pathway, previously characterized by reverse genetic in ... [more ▼]

In photosystem I (PSI), phylloquinone participates to electron transfer as secondary electron acceptor (A1). The phylloquinone biosynthesis pathway, previously characterized by reverse genetic in Synechocystis sp. PCC 6803, involves 8 enzymatic steps from chorismate [1]. In the green alga Chlamydomonas reinhardtii, characterization of phylloquinone biosynthesis was still partial and only one mutant deficient for MEND was characterized [2]. In the present work, we found MENA-H homologs in C. reinhardtii genomic database. In particular, MENF, MEND, MENC, and MENH catalytic domains are present in a single ORF (named PHYLLO by similarity to gene organisation in Arabidopsis). We then took advantage of the fact that a double reduction of plastoquinone (PQ) in PQH2 occurs in anoxia into the A1 site in the mend mutant, interrupting photosynthetic electron transfer [3], to isolate new phylloquinone-deficient strains. UPLC-MS analysis confirmed the absence of phylloquinone in four news mutants impaired in MENA, MENB, MENC (PHYLLO) and MENE. Despite this loss, men mutants are still able to grow in low light but are high light-sensitive. In low light, the level of active PSII in men mutants is identical to that of the wild-type, but the level of active PSI is reduced by 30-40% as assayed by spectroscopic measurements. This decrease is more pronounced when cells are exposed to high light intensities during 4 hours. The level of active PSI is ~ 10% of wild-type cells and the electron photosynthetic transfer is reduced accordingly. Reorganization of the photosynthetic apparatus following lack of phylloquinone in men mutants is discussed. [less ▲]

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See detailIsolation and characterization of mutants deficient in four steps of the phylloquinone biosynthesis pathway in Chlamydomonas reinhardtii.
Emonds-Alt, Barbara ULg; Remacle, Claire ULg; Cardol, Pierre ULg

Poster (2016, April 26)

In photosystem I (PSI), phylloquinone participates to electron transfer as secondary electron acceptor (A1). The phylloquinone biosynthesis pathway, previously characterized by reverse genetic in ... [more ▼]

In photosystem I (PSI), phylloquinone participates to electron transfer as secondary electron acceptor (A1). The phylloquinone biosynthesis pathway, previously characterized by reverse genetic in Synechocystis sp. PCC 6803, involves 8 enzymatic steps from chorismate [1]. In the green alga Chlamydomonas reinhardtii, characterization of phylloquinone biosynthesis was still partial and only one mutant deficient for MEND was characterized [2]. In the present work, we found MENA-H homologs in C. reinhardtii genomic database. In particular, MENF, MEND, MENC, and MENH catalytic domains are present in a single ORF (named PHYLLO by similarity to gene organisation in Arabidopsis). We then took advantage of the fact that a double reduction of plastoquinone (PQ) in PQH2 occurs in anoxia into the A1 site in the mend mutant, interrupting photosynthetic electron transfer [3], to isolate new phylloquinone-deficient strains. UPLC-MS analysis confirmed the absence of phylloquinone in four news mutants impaired in MENA, MENB, MENC (PHYLLO) and MENE. Despite this loss, men mutants are still able to grow in low light but are high light-sensitive. In low light, the level of active PSII in men mutants is identical to that of the wild-type, but the level of active PSI is reduced by 30-40% as assayed by spectroscopic measurements. This decrease is more pronounced when cells are exposed to high light intensities during 4 hours. The level of active PSI is ~ 10% of wild-type cells and the electron photosynthetic transfer is reduced accordingly. Reorganization of the photosynthetic apparatus following lack of phylloquinone in men mutants is discussed. [less ▲]

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See detailIsolation and characterization of photoactive complexes of NADPH : protochlorophyllide oxidoreductase from wheat
Chahdi, M. A. O.; Schoefs, B.; Franck, Fabrice ULg

in Planta (1998), 206(4), 673-680

A photoactive substrate-enzyme complex of the NADPH:protochlorophyllide oxidoreductase (POR; EC 1.3.1.33) was purified from etiolated Triticum aestivum L. by gel chromatography after solubilization of ... [more ▼]

A photoactive substrate-enzyme complex of the NADPH:protochlorophyllide oxidoreductase (POR; EC 1.3.1.33) was purified from etiolated Triticum aestivum L. by gel chromatography after solubilization of prolamellar bodies by dodecyl-maltoside. Irradiation by a 1-ms flash induced the phototransformation of protocholorophyllide a (Pchlide) with -196 degrees C absorbance and emission maxima at 640 and 643 nm, respectively. The apparent molecular weight of this complex was 112 +/- 24 kDa, which indicates aggregation of enzyme subunits. By lowering the detergent concentration in the elution buffer, a 1080 +/- 250-kDa particle was obtained which displayed the spectral properties of the predominant form of photoactive Pchlide in vivo (-196 degrees C absorbance and fluorescence maxima at 650 and 653 nm). In this complex, FOR was the dominant polypeptide. Gel chromatography in the same conditions of an irradiated sample of solubilized prolamellar bodies indicated rapid disaggregation of the complex after Pchlide phototransformation. High performance liquid chromatographic analysis of the FOR complexes obtained using two detergent concentrations indicates a possible association of zeaxanthin and violaxanthin with the photoactive complex. [less ▲]

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See detailIsolation and characterization of preantral follicles from fetal bovine ovaries
Hulshof, S. C. J.; Figueiredo, José Ricardo; Beckers, Jean-François ULg et al

in Veterinary Quarterly (The) (1994), 16(2), 78-80

A simple, mechanical method is described for the isolation of preantral follicles bovine foetuses of 220-280 days of gestation. On average, 2918+621 (s.d.) preantral follicles were isolated per ovary. The ... [more ▼]

A simple, mechanical method is described for the isolation of preantral follicles bovine foetuses of 220-280 days of gestation. On average, 2918+621 (s.d.) preantral follicles were isolated per ovary. The isolated preantral follicles were characterized on the basis of the morphological appearance of the surrounding granulosa cells, the number of granulosa cell layers, and their diameter. The results show that primordial, primary, and secondary follicles differ morphologically and that they can be classified by their diameter. [less ▲]

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See detailIsolation and characterization of respiratory NADH:ubiquinone oxidoreductase (complex I) mutants in Chlamydomonas reinhardtii
Remacle, Claire ULg; Baurain, Denis ULg; Colin, Martine et al

in Archives of Physiology & Biochemistry (2000), 108(Supplement 1), 10

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See detailIsolation and characterization of the human prolactin gene
Truong, Anh T; Duez, Colette ULg; Belayew, Alexandra et al

in EMBO Journal (1984), 3(2), 429-37

Prolactin (PRL) and growth hormone (GH) genes derive from a common ancestor and still share some sequence homologies. Their expression in the pituitary gland is regulated in opposite directions by most of ... [more ▼]

Prolactin (PRL) and growth hormone (GH) genes derive from a common ancestor and still share some sequence homologies. Their expression in the pituitary gland is regulated in opposite directions by most of the many hormones acting on them. This provides an interesting system to study sequences involved in gene expression. Using a human PRL cDNA clone as a probe, we screened a human genomic DNA library in lambda phage and isolated a single recombinant comprising the whole hPRL gene. It was characterized by restriction endonuclease mapping and cDNA hybridization, by DNA heteroduplex analysis and by nucleotide sequencing. The hPRL gene is present as a single copy per haploid genome, is approximately 10 kb long and contains four introns, three of which interrupt the coding sequence at the same locations as in the known GH and PRL genes. The origin of transcription was determined by S1 mapping on prolactinoma mRNAs. The search for direct and inverted repeats, as well as dyad symmetries was carried out in the 900-bp sequenced in the 5'-flanking region. Sequence homologies between hPRL, hGH and rPRL were derived from computer drawn matrices for these upstream regions. [less ▲]

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See detailIsolation and characterization of Ts19 Fragment II, a new long-chainpotassium channel toxin from Tityus serrulatus venom
Cerni, Felipe Augusto; Pucca, Manuela Berto; Amorim, Fernanda Gobbi et al

in Peptides (2016), 80

Ts19 Fragment II (Ts19 Frag-II) was first isolated from the venom of the scorpion Tityus serrulatus (Ts). It is aprotein presenting 49 amino acid residues, three disulfide bridges, Mr5534 Da and was ... [more ▼]

Ts19 Fragment II (Ts19 Frag-II) was first isolated from the venom of the scorpion Tityus serrulatus (Ts). It is aprotein presenting 49 amino acid residues, three disulfide bridges, Mr5534 Da and was classified as a newmember of class (subfamily) 2 of the -KTxs, the second one described for Ts scorpion. The -KTx familyis composed by two-domain peptides: N-terminal helical domain (NHD), with cytolytic activity, and aC-terminal CS domain (CCD), with Kv blocking activity. The extensive electrophysiological screening(16 Kv channels and 5 Nav channels) showed that Ts19 Frag-II presents a specific and significant blockingeffect on Kv1.2 (IC50value of 544 ± 32 nM). However, no cytolytic activity was observed with this toxin.We conclude that the absence of 9 amino acid residues from the N-terminal sequence (compared to Ts19Frag-I) is responsible for the absence of cytolytic activity. In order to prove this hypothesis, we synthesizedthe peptide with these 9 amino acid residues, called Ts19 Frag-III. As expected, Ts19 Frag-III showed tobe cytolytic and did not block the Kv1.2 channel. The post-translational modifications of Ts19 and itsfragments (I–III) are also discussed here. A mechanism of post-translational processing (post-splitting) issuggested to explain Ts19 fragments production. In addition to the discovery of this new toxin, this reportprovides further evidence for the existence of several compounds in the scorpion venom contributing tothe diversity of the venom arsenal. [less ▲]

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See detailIsolation And Chemical Evaluation Of Carob (Ceratonia Siliqua L.) Seed Germ
Dakia, Patrick; Wathelet, Bernard ULg; Paquot, Michel ULg

in Food Chemistry (2007), 102(4),

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See detailIsolation and Cultivation of a Xylanolytic Bacillus subtilis Extracted from the Gut of the Termite Reticulitermes santonensis
Tarayre, Cédric ULg; Brognaux, Alison ULg; Brasseur, Catherine ULg et al

in Applied Biochemistry and Biotechnology (2013)

The aim of this work was the isolation of xylanolytic microorganisms from the digestive tract of the termite Reticulitermes santonensis. The reducing sugars released after the hydrolysis of xylans can be ... [more ▼]

The aim of this work was the isolation of xylanolytic microorganisms from the digestive tract of the termite Reticulitermes santonensis. The reducing sugars released after the hydrolysis of xylans can be further fermented to provide bioethanol. A xylanolytic strain of Bacillus subtilis was isolated from the hindgut of the termite and displayed amylase and xylanase activities. The bacterium was grown on media containing agricultural residues: wheat bran, wheat distiller’s grains, and rapeseed oil cake. Wheat bran led to the highest induction of xylanase activity, although the development of the strain was less fast than in the other media. It was possible to reach maximal xylanase activities of 44.3, 33.5, and 29.1 I.U./ml in the media containing wheat bran, wheat distiller’s grains, and rapeseed oil cake, respectively. Mass spectrometry identified a wide range of xylose oligomers, highlighting an endoxylanase activity. The enzyme was stable up to 45 °C and displayed an optimal pH close to 8. [less ▲]

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See detailIsolation and cultivation of cellulolytic and xylanolytic bacteria and molds extracted from the gut of the termite Reticulitermes santonensis (3DV.1.14)
Tarayre, Cédric ULg; Bauwens, Julien ULg; Mattéotti, Christel et al

Poster (2013, June)

Biofuel production can be based on the use of agro-residues, consisting in a complex lignocellulosic structure which is not easily hydrolysable. The digestive tract of the termite Reticulitermes ... [more ▼]

Biofuel production can be based on the use of agro-residues, consisting in a complex lignocellulosic structure which is not easily hydrolysable. The digestive tract of the termite Reticulitermes santonensis contains a diversified microflora able to hydrolyze the wood components. Bacteria, molds and protists form efficient consortia, able to break the lignocellulosic complex by producing enzymes, such as xylanases and cellulases. Our purpose is the isolation of microbial strains from termite guts in order to evaluate their potential for hydrolysis of lignocellulosic materials. Termites were fed using different diets chosen to improve the xylanolytic and cellulolytic microflora: wood, microcristalline cellulose (added with lignin or not), α-cellulose (added with lignin or not) and birchwood xylan. Then, dissections were realized to isolate the potential xylanolytic and cellulolytic strains. This approach led us to isolate and to study several strains of bacteria (Bacillus sp. strain CTGx and Chryseobacterium sp. strain CTGx) and molds (Trichoderma virens strain CTGx and Sarocladium kiliense strain CTGx). These microorganisms were able to hydrolyze starch, xylan, cellulose, carboxymethylcellulose, esculin, β-glucan and Whatman® filter paper. They can produce glucose and xylose monomers and oligomers which can be further fermented to produce bioethanol. [less ▲]

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See detailIsolation and cultivation of xylanolytic and cellulolytic Sarocladium kiliense and Trichoderma virens from the gut of the termite Reticulitermes santonensis
Tarayre, Cédric ULg; Bauwens, Julien ULg; Brasseur, Catherine ULg et al

in Environmental Science and Pollution Research (2014)

The purpose of this work was the isolation and cultivation of cellulolytic and xylanolytic microorganisms extracted from the gut of the lower termite Reticulitermes santonensis. Microcrystalline cellulose ... [more ▼]

The purpose of this work was the isolation and cultivation of cellulolytic and xylanolytic microorganisms extracted from the gut of the lower termite Reticulitermes santonensis. Microcrystalline cellulose (with and without lignin) and beech wood xylan were used as diets instead of poplar wood in order to select cellulose and hemicellulose-degrading fungi. The strain Sarocladium kiliense (Acremonium kiliense) CTGxxyl was isolated from the termites fed on xylan, while the strain Trichoderma virens CTGxAviL was isolated from the termites fed on cellulose (with and without lignin). Both molds were cultivated in liquid media containing different substrates: agro-residues or purified polymers. S. kiliense produced maximal β-glucosidase, endo-1,4-β-D-glucanase, exo-1,4-β-D-glucanase and endo-1,4-β-D-xylanase activities of 0.103, 3.99, 0.53, and 40.8 IU/ml, respectively. T. virens produced maximal β-xylosidase, endo-1,4-β-D-glucanase, exo-1,4-β-D-glucanase, and endo-1,4-β-D-xylanase activities of 0.38, 1.48, 0.69, and 426 IU/ml. The cellulase and the xylanase of S. kiliense, less common than T. virens, were further investigated. The optimal activity of the xylanase was observed at pH 9–10 at 60 °C. The cellulase showed its maximal activity at pH 10, 70 °C. Zymography identified different xylanases produced by both molds, and some fragment sizes were highlighted: 35, 100, and 170 kDa for S. kiliense and 20, 40, 80, and 170 kDa for T. virens. In both cases, endo-1,4-β-D-xylanase activitieswere confirmed through mass spectrometry. [less ▲]

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