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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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Peer Reviewed
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 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 ▲]

Detailed reference viewed: 35 (11 ULg)