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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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See detailIsolation and evaluation of bacteria and fungi as biological control agents against Rhizoctonia solani.
Lahlali, R.; Bajii, M.; Jijakli, Mohamed ULg

in Communications in agricultural and applied biological sciences (2007), 72(4), 973-982

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See detailIsolation and identification of a new Bacillus strain for amylase production
Bakri, Y.; Ammouneh, H.; El-Khouri, S. et al

in Research in Biotechnology (2012), 3(6), 51-58

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See detailIsolation and identification of a new fungal strain for amylase biosynthesis
Bakri, Y.; Masson, M.; Thonart, Philippe ULg

in Polish Journal of Microbiology (2009), 58(3), 269-273

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See detailIsolation and identification of anthropogenic particles in fish stomachs by Raman spectroscopy: a new method
Collard, France ULg; Gilbert, Bernard ULg; Eppe, Gauthier ULg et al

in Organohalogen Compounds (2015, August)

Microplastic particles (MP) contaminate oceans and affect marine organisms in several ways. Ingestion combined with food intake is generally reported. However, data interpretation is often circumvented by ... [more ▼]

Microplastic particles (MP) contaminate oceans and affect marine organisms in several ways. Ingestion combined with food intake is generally reported. However, data interpretation is often circumvented by the difficulty to separate MP from bulk samples. Visual examination is often used as one or the only step to sort these particles. However, color, size and shape are insufficient and often unreliable criteria. Here we present an isolation method of MP specially adapted to a subsequent analysis by Raman spectroscopy. This method avoids fluorescence problems allowing the identification anthropogenic particles (AP) from stomach contents of fish by Raman spectroscopy. It was validated with commercial samples of microplastics and cotton along with stomach contents from three different Clupeiformes fishes: Clupea harengus, Sardina pilchardus and Engraulis encrasicolus. The optimized digestion and isolation protocol showed no visible impact on microplastics and cotton particles while the spectroscopic analysis allowed precise identification of microplastics and textile fibers. This approach allowed us to isolate 35 particles. These were analyzed by Raman spectroscopy: eleven were microplastics and thirteen were made of cellulose or lignin, or both (mostly fibers). Some particles were not identified but contained artificial colorants. This isolation protocol will help to assess the presence, quantity and composition of AP in planktivorous fish stomachs. [less ▲]

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See detailIsolation and identification of antioxidant phytochemicals from Cuban species of the genera Erythroxylum P. Browne and Pluchea Cass
Perera Cordova, Wilmer ULg

Doctoral thesis (2012)

Phytochemicals showing antioxidant properties are largely recognized as beneficial to human health and disease prevention. Cuba is well known for having a rich flora with a high percentage of endemic ... [more ▼]

Phytochemicals showing antioxidant properties are largely recognized as beneficial to human health and disease prevention. Cuba is well known for having a rich flora with a high percentage of endemic species; it is considered as an interesting source of plant species for searching bioactive metabolites. Polar and non-polar extracts were prepared by fractionation from macerations in ethanol: H2O (7:3 v/v) from several Cuban plant species. The antioxidant capacity and the concentration in phenolic compounds were assayed in these extracts. The native species Pluchea carolinensis (Jacq.) G. Don., Pluchea odorata (L.) Cass. and Pluchea rosea Godfrey were identified as the most promising sources of antioxidants. The n-butanol extracts obtained by reflux from endemic Erythroxylum alaternifolium A. Rich. var. alaternifolium, var. parvifolium and suborbiculare also displayed high antioxidant capacity and high levels in phenolic compounds. The antioxidant capacity and the phenolic content of hydroalcoholic macerations from leaves, inflorescences, stems and roots of species of the genus Pluchea were also compared. Leaf extracts followed by inflorescence extracts showed the highest values of antioxidant capacity. The species P. carolinensis was used as a model to evaluate the antioxidant capacity in two locations and two phenological stages and also for monitoring the antioxidant capacity over some months. Natural adult specimens presented a higher phenolic content, as well as higher antioxidant capacity than young and cultivated specimens. Maximal antioxidant capacity and concentrations in phenolics were recorded in January, September and December; over the blooming stage (March), the antioxidant capacity was minimal. Two different solvents of extraction and two methods were also screened for extracting the maximal concentrations in antioxidants. Hydroalcoholic extractions showed higher antioxidant capacity than ethanolic ones. However, no difference in antioxidant capacity were measured between the two methods of extraction. The antioxidant capacity of P. carolinensis was also measured in leaves of plantlets micro-propagated on three different in vitro culture media and it was compared with leaves of young specimens grown ex-vitro. The inclusion of a cytokinin in culture media increased the antioxidant capacity of the leaf extracts of plantlets grown in vitro. However, the antioxidant capacity of plantlets grown in vitro was lower than that of young specimens grown ex-vitro. Additionally, several phenolic compounds were isolated from the species studied. Two flavonol glycosides were isolated from n-butanol leaf extract of Erythroxylum alaternifolium var. alaternifolium. Moreover, 22 phenolic compounds (including 9 phenolic acids and 13 flavonoids) were also identified from leaf, inflorescence and stem extracts of the three Pluchea species. All of them are reported for the first time in Cuban Pluchea species. In addition, rosmarinic acid, ferulic acid, quercetagetin, herbacetin, quercitrin, eupalitin and 3-methyl-quercetagetin were described for the first time in the genus Pluchea. The antioxidant capacity of each of these compounds was evaluated. The results suggested that most of these phytochemicals have an important contribution to the antioxidant capacity found in plant extracts. [less ▲]

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See detailIsolation and identification of dominant osmophilic Leuconostoc strains from traditional date product “Btana”
Abekhti, Abdelkader; Daube, Georges ULg; Kihal, M.

in International Food Research Journal [=IFRJ] (2014), 21(4), 1261-1268

The current study aimed to isolate and identify dominant osmophilic bacteria associated with a traditional date product named “Btana”, produced in south region of Maghreb countries. Samples were randomly ... [more ▼]

The current study aimed to isolate and identify dominant osmophilic bacteria associated with a traditional date product named “Btana”, produced in south region of Maghreb countries. Samples were randomly collected after two month of storage from tow villages (Mtarfat and Abani) in the Algerian southern department “Adrar”. A high osmotic pressure medium (MSE) was used for isolation of osmophilic bacteria, which were purified and examined for macroscopic and microscopic shape, Gram stain, catatalse, oxydase, acetoine and ADH production, reduction of nitrate, and motility. Isolates were then subculture on MRS medium for production of dextran, gas from glucose, growth in the presence of NaCl (3, 6.5 %) and sucrose (10, 20, 30, 40, and 50 %), pH tolerance (4.8, 6.5), growth temperature (10, 37, and 45°C) and thermo resistance (55°C for 15 min), enzymatic activity (proteolytic, lipolytic, hemolysis). Isolates were identified to specie’s level by sugar fermentation. Their growth and acidification kinetic were also studied. Results identified two species of Leuconostoc; Leuconostoc mesenteroides subsp. mesenteroides and Leuconostoc mesenteroides subsp. dextranicum. They show a high antibacterial activity against four indicator bacteria; Pseudomonas aeruginosa ATCC 27853, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. [less ▲]

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See detailIsolation and identification of inulooligosaccharides resulting from inulin hydrolysis
Ronkart, Sebastien N; Blecker, Christophe ULg; Fourmanoir, Hélène et al

in Analytica Chimica Acta (2007), 604(1), 81-87

In this study, inulooligosaccharides (F-n-type inulin) resulting from the endo-inulinase hydrolysis of globe artichoke inulin were purified and characterized. The aim was to produce F-n oligomer standards ... [more ▼]

In this study, inulooligosaccharides (F-n-type inulin) resulting from the endo-inulinase hydrolysis of globe artichoke inulin were purified and characterized. The aim was to produce F-n oligomer standards with the intention of identifying them in the complex inulin chromatogram. Inulin was extracted from globe artichoke and presented a high average degree of polymerization (DP) of about 80 as determined by high-performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD). This inulin was hydrolyzed by a commercial endo-inulinase yielding a product with a very high F-n/GF(n), molecule ratio, thus limiting the interference of GF(n) during the purification process. High performance size exclusion chromatography was used to individually isolate and collect each retention peak corresponding to a specific oligomer. The purity of these fractions was checked by HPAEC-PAD and showed that relatively pure molecules were produced. Electrospray ionization mass spectrometry allowed the molecular weight determination of these purified oligomers and ascertained their DP as F-2, F-3 and F-4. These F2-4 standards were used with glucose, fructose, sucrose and GF(2-4) (commercially available) to spike commercial oligofructose products in order to determine the elution profile in the HPAEC-PAD chromatogram. [less ▲]

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See detailIsolation and identification of potential antimalarial compounds from endemic plants of Reunion Island
Bordignon, Annélise ULg; Cieckiewicz, Ewa ULg; Jansen, Olivia ULg et al

Poster (2015, July 16)

Malaria is known as the most important parasitic disease around the world with 584 000 malaria deaths worldwide in 2013 [1]. Due to the problem of increased parasite resistance, natural products from ... [more ▼]

Malaria is known as the most important parasitic disease around the world with 584 000 malaria deaths worldwide in 2013 [1]. Due to the problem of increased parasite resistance, natural products from endemic plants of Reunion Island, hot spot of promising biodiversity, could represent an important source of new antimalarial drugs. The aim of this thesis research focuses on the evaluation of potential antiplasmodial activity of medicinal plants from Reunion Island. A global screening of plants extracts from Reunion Island was performed on Plasmodium falciparum 3D7 chloroquine-sensitive strain revealed by colorimetric method as described in previous reports [2]. Monimia rotundifolia was then selected due to its promising in vitro activity against Plasmodium. Bioguided fractionation was realized using Prep HPLC techniques and led to the isolation of aporphine-type alkaloids from Monimia rotundifolia leaves dichloromethane extract. Further investigations are in process to confirm the antiplasmodial activities of these alkaloids and to determine their structures. References: [1] WHO, World Malaria report 2014. [2] Jansen O. et al., Evaluation of 13 selected medicinal plants from Burkina Faso for their antiplasmodial properties. J Ethnopharmacol 2010, 130:143-150. [less ▲]

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See detailIsolation and Identification of Steroid and Flavonoid Glycosides from the Flowers of Allium gramineum
Mskhiladze, Lasha; Chincharadze, David; St-Gelais, Alexis et al

in International Journal of Pharmaceutical Sciences and Drug Research (2016), 8(4), 240-243

The isolation and identification of β-sitosterol 3-O-β-glucopyranoside, quercetin 3-O-β-glucopyranoside, isorhamnetin 3,4'-di-O-β-glucopyranosideand isorhamnetin 3,7-di-O-β-glucopyranosidefrom the flowers ... [more ▼]

The isolation and identification of β-sitosterol 3-O-β-glucopyranoside, quercetin 3-O-β-glucopyranoside, isorhamnetin 3,4'-di-O-β-glucopyranosideand isorhamnetin 3,7-di-O-β-glucopyranosidefrom the flowers of Allium gramineumthat growsin Georgia.The structures of isolated compounds havebeen determined by 1H and 13C nuclear magnetic resonance (NMR) Isolation and Identification of Steroid and Flavonoid Glycosides from the Flowers of Allium gramineum. Available from: https://www.researchgate.net/publication/305851744_Isolation_and_Identification_of_Steroid_and_Flavonoid_Glycosides_from_the_Flowers_of_Allium_gramineum [accessed Aug 5, 2016]. [less ▲]

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See detailIsolation and identification of two new fungal strains for xylanase production
Bakri, Y.; Masson, M.; Thonart, Philippe ULg

in Applied Biochemistry and Biotechnology (2010), 162(6), 1626-1634

Fungi are well known for their ability to excrete enzymes into the environment. The aim of this work was to evaluate xylanase production by fungi isolated from soil. One hundred and thirty-six fungal ... [more ▼]

Fungi are well known for their ability to excrete enzymes into the environment. The aim of this work was to evaluate xylanase production by fungi isolated from soil. One hundred and thirty-six fungal isolates were screened for xylanase production. Two xylanase producing isolates, FSS117 and FSS129, were identified on the basis of analyses of 5,8S gene sequencing. The closest phylogenetic neighbors according to 5,8S gene sequence data for the two isolates were Aspergillus tubingensis and Aspergillus terreus, respectively. When birchwood xylan or corn cob hulls was used as a substrate for 5 days under submerged culture cultivation, xylanase production from A. terreus FSS129 was 113 and 174 IU ml -1, respectively. The pH and temperature for optimum xylanase activity were 8 and 65∈°C. © 2010 Springer Science+Business Media, LLC. [less ▲]

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See detailIsolation and long-term cultivation of human tonsil follicular dendritic cells.
Tsunoda, R.; Nakayama, M.; Onozaki, K. et al

in Virchows Archiv. B : Cell pathology (1990), 59(2), 95-105

Highly purified follicular dendritic cells (FDC) were isolated from human tonsils and cultivated for up to 150 days. The cell separation method employed produced pure aggregates (FDC-clusters) composed of ... [more ▼]

Highly purified follicular dendritic cells (FDC) were isolated from human tonsils and cultivated for up to 150 days. The cell separation method employed produced pure aggregates (FDC-clusters) composed of FDC and germinal center lymphoid cells, useful for the analysis of the relationship between these two cell types and of the behavior of FDC in culture. During the first few days of culture, lymphoid cells located between FDC extensions survived better than those which were free or partly covered by FDC. After 6 days, the lymphoid population degenerated and only the FDC survived. The unique antigenic pattern of FDC (positive for HLA-DR. DRC-1, CD14b, CD21, CD23, CD35) disappeared within a few days of culture. Recombinant interferon-gamma exerted a positive effect either on retaining HLA-DR expression or on the reexpression of these antigens by FDC. HLA-ABC antigens were traced until the 10th day and desmosomal junctions until the 14th day. Subsequently, FDC presented peculiar features, including oval and rhomboid shapes, one to ten nuclei, fine amoeboid extensions, stress fibers and a radical dense zone in their cytoplasm. FDC possessed actin, tubulin and vimentin, but neither desmin nor cytokeratin. After 40 days of culture, FDC enlarged and were covered with abundant membrane extensions. Even when kept as long as 150 days in vitro. FDC did not proliferate in any of the culture conditions employed. [less ▲]

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See detailIsolation and Partial Characterization of a Pregnancy-Associated Glycoprotein Family from the Goat Placenta
Garbayo, Juana Maria; Remy, Benoit; Alabart, José Luis et al

in Biology of Reproduction (1998), 58(1), 109-115

Antigen(s) immunologically related to pregnancy-associated glycoproteins (PAGs) have previously been detected in the serum of pregnant goats. In this work, we describe a partial characterization of a ... [more ▼]

Antigen(s) immunologically related to pregnancy-associated glycoproteins (PAGs) have previously been detected in the serum of pregnant goats. In this work, we describe a partial characterization of a family of PAGs isolated from the placenta of the goat. The procedure, monitored by RIA, included extraction of proteins at neutral pH, acidic, and ammonium sulfate precipitations; and gel filtration and ion exchange chromatographies. Immunoreactivity, initially located in the acidic supernatant and in the 40-80% ammonium sulfate fractions, was equally apportioned between the 0.04 and 0.08 M NaCl DEAE fractions. After further purification of both DEAE fractions, the preparations were subjected to one- and two-dimensional electrophoresis, and individual polypeptides were analyzed by amino acid sequencing. Three PAGs, which differed in amino acid sequence and apparent molecular masses (62, 59, and 55 kDa), were detected, each containing several isoforms with different pls: caprine (c) PAG62 (pl: 5.1, 4.8), cPAG59 (pl: 6.2, 5.9, 5.6), and cPAG55 (pl: 5.3, 5.1, 4.9). These proteins had high sequence identities to each other and to PAGs purified from other species. Each had two putative N-glycosylation sites within the 27 amino terminal residues sequenced. This work demonstrates that PAGs are present in goat placenta and that multiple forms are expressed. [less ▲]

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