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See detailIn vitro antitrypanosomal and antiplasmodial activities of crude extracts and essential oils of Ocimum gratissimum Linn from Benin and influence of vegetative stage.
Kpadonou Kpoviessi, Benedicta G. H.; Kpoviessi, Salome D. S.; Yayi Ladekan, Eleonore et al

in Journal of ethnopharmacology (2014), 155(3), 1417-23

ETHNOPHARMACOLOGICAL RELEVANCE: Different parts of Ocimum gratissimum Linn are largely used in folk medicine for the treatment of many diseases, some of which related to parasitical infections as fevers ... [more ▼]

ETHNOPHARMACOLOGICAL RELEVANCE: Different parts of Ocimum gratissimum Linn are largely used in folk medicine for the treatment of many diseases, some of which related to parasitical infections as fevers and headaches. In order to validate their use and to clarify the plant part which possesses the best antiparasitic properties, we decided to evaluate the in vitro antiplasmodial and antitrypanosomal activities of essential oils and crude extracts from leaves, stems and seeds of Ocimum gratissimum as well as their cytotoxicity. MATERIALS AND METHODS: The essential oils and ethanol crude extracts of leaves and stems of Ocimum gratissimum from Benin, were obtained in pre and full flowering stages. Seeds obtained only in full flowering stage, were also extracted. The oils were isolated by hydrodistillation and analyzed by GC/MS and GC/FID. Extracts and essential oils were tested in vitro against Trypanosoma brucei brucei and Plasmodium falciparum. Cytotoxicity was evaluated in vitro against Chinese Hamster Ovary (CHO) cells and the human non cancer fibroblast cell line (WI38) through MTT assay to evaluate the selectivity and toxicity was assessed against Artemia salina Leach. RESULTS: The essential oils and non-volatile crude extracts of Ocimum gratissimum were more active on Trypanosoma brucei brucei than on Plasmodium falciparum (3D7). This activity varies according to the vegetative stage (pre and full flowering) and the plant part (seeds, stems and leaves) extracted. The best growth inhibition of Trypanosoma brucei brucei was observed with ethanol crude extracts of leaves (IC50=1.66 +/- 0.48 mug/mL) and seeds (IC50=1.29 +/- 0.42 mug/mL) in full flowering stage with good selectivity (SI>10). The chemical composition of the essential oil from aerial parts (47 compounds), characterized by the presence as main constituents of p-cymene, thymol, gamma-terpinene, beta-myrcene and alpha-thujene, depends on the vegetative stage. The oil contained some minor compounds such as myrcene (IC50=2.24 +/- 0.27mug/mL), citronellal (IC50=2.76 +/- 1.55mug/mL), limonene (IC50=4.24 +/- 2.27mug/mL), with good antitrypanosomal activities. These oils and crude extracts were not toxic against Artemia salina Leach and had a low cytotoxicity except leaves and seeds ethanol extracts obtained in full flowering which showed toxicity against CHO and WI38 cells. CONCLUSIONS: Our study shows that ethanol crude extracts of leaves and seeds of Ocimum gratissimum in full flowering stage can be a good source of antitrypanosomal agents. This is the first report about the relation between the plant part extracted, the vegetative stage of the plant, the antitrypanosomal and antiplasmodial activities and the cytotoxicity of essential oils and non-volatile extracts of Ocimum gratissimum from Benin. [less ▲]

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See detailIn vitro approach to study the synergistic effects of tobramycin and clarithromycin against Pseudomonas aeruginosa biofilms using prokaryotic or eukaryotic culture media
Thellin, Olivier ULg; Zorzi, Willy ULg; Jolois, Olivier et al

in International Journal of Antimicrobial Agents Corresponding (in press)

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See detailIn vitro assessment of novel carbohydrates prebiotic potential in a co-inoculation model of the pig intestines
Tran, Thi Hanh Tham ULg; Boudry, Christelle; Everaert, Nadia ULg et al

in Book of Abstracts (2015)

Introduction: Innovative plant biomass fractionation methods produce new feed additives that could modulate the pig intestinal microbial population. Material and methods: Such novel indigestible ... [more ▼]

Introduction: Innovative plant biomass fractionation methods produce new feed additives that could modulate the pig intestinal microbial population. Material and methods: Such novel indigestible carbohydrates (CHO) were investigated for their prebiotic potential and their influence on Salmonella thyphimurium in a co-inocu¬lation in vitro fermentation model of the pig intestines. Inulin, cellobiose, pecto- (POS), iso-malto- (IMOS), xylo-oligosaccharides (XOS), and gluconic acid (GLU) were fermented by fecal microbes for 72h. Salmonella (7.3 log CFU/ml) were co-inoculated after 6h . Fer¬mentation kinetics was modeled and after 6, 12, 24 h, broth was analysed for short-chain fatty acid using HPLC and bacterial population using q-PCR. Results and discussion: Cellobiose was the fastest fermenting CHO followed by inulin and IMOS (P<0.01). After 6h, cellobiose yielded the highest SCFA production (684 mg/g) and lactate molar ratio (0.484). POS fermented slower. XOS and GLU were little fermented (150 and 175 mg SCFA/g after 24h). Nonetheless, GLU yielded the highest butyrate molar ratio (0.290 at 12h) (P<0.01). Although Salmonella counts did not differ, some CHO dis¬played obvious prebiotic properties, namely inulin and IMOS since they supported the highest growth of Lactobacilli and Bifidobacteria populations after 6 and 12h of fermen¬tation (8.18 to 8.56 log CFU/ml) (P<0.01). Cellobiose and GLU scored well for Lactobacilli too, but poorly for Bifidobacteria (6.41 to 6.92 log CFU/ml) (P<0.01). It is concluded that IMOS seem the most promising prebiotic but owing to their fermentation patterns yield¬ing high levels of lactate or butyrate, also cellobiose and GLU deserve further investigation in in vivo models. [less ▲]

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See detailIn vitro Biocompatibility and Genotoxicity Assessment of a Gentamicin-Loaded Monoolein Gel Intended to Treat of Chronic Osteomyelitis
Ouédraogo, M; Nacoulma, E.C; Semdé, R et al

in Journal of Pharmacology & Toxicology (2008), 3(5),

The aim of the study was to assess in vitro the biocompatibility and the genotoxicity of a gentamicin-loaded monoolein gel intended to treat of chronic osteomyelitis. Indeed, we are developing ... [more ▼]

The aim of the study was to assess in vitro the biocompatibility and the genotoxicity of a gentamicin-loaded monoolein gel intended to treat of chronic osteomyelitis. Indeed, we are developing biodegradable implants based on monoolein and gentamicin. The results of formulations, physico-chemical characterization of the formulated implants and in vitro release kinetic of gentamicin from implants were encouraging. As biocompatibility and absence of genotoxicity are the prerequisites for safe use of implants, we performed in vitro hemolysis, cytotoxicity and, genotoxicity tests. Hemolysis was evaluated by incubating human erythrocytes in direct contact with the implant whereas cytotoxicity was evaluated by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay using fibroblasts and macrophages. Alkaline comet Assay was used to evaluate genotoxic potential of the implants. From these in vitro assays, the implant based on monoolein and gentamicin showed no genotoxic potential and has satisfactory biocompatibility. [less ▲]

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See detailIn vitro characterisation of dietary fibre fermentation in the pig intestines and its influence on nitrogen excretion
Bindelle, Jérôme ULg

Doctoral thesis (2008)

Increasing attention has been paid to dietary fibre (DF) fermentation in the large intestine of pigs during the past years in pig nutrition. The bacterial growth supported by DF intestinal fermentation ... [more ▼]

Increasing attention has been paid to dietary fibre (DF) fermentation in the large intestine of pigs during the past years in pig nutrition. The bacterial growth supported by DF intestinal fermentation induces a shift of N excretion from urea in urine to bacterial protein in faeces that reduces NH3 emission from the manure. The objective of this thesis was to investigate the relationship between DF fermentability, intestinal bacteria growth and the N excretion. In the first part, an in vitro gas-test method using a living bacterial inoculum developed for ruminants was adapted to the pig. The use of pig colonic content was compared to faeces for the preparation of the inoculum and it was concluded that faeces could replace intestinal content, avoiding the use of cannulated animals. Secondly, the influence of a pespin-pancreatin hydrolysis prior to the fermentation in order to simulate digestion in the stomach and the small intestine was demonstrated. Finally, the influence of the faeces donnor bodyweight and the dietary fibre content of its diet on the gas production kinetics was shown. When studying a topic related to a specific category of pig, it is recommended to use animals from the same category as faeces donors to prepare the inoculum. In the second part of the thesis, the amount of protein synthesis (PS) by faecal microbes fermenting different sources of purified carbohydrates, or ingredients differing in DF content, was measured using 15N-labelled NH4Cl in the inoculum. PS ranged between 9.8 and 22.9 mg N g-1 fermented carbohydrate according to the rate of fermentation of the carbohydrate and its soluble fibre content. These in vitro observations were confirmed through in vivo experiments with diets containing increasing levels of soluble DF: in vitro PS passed from 1.51 to 2.35 mg N g-1 diet while in vivo urinary- N:fecal–N excretion ratio decreased from 2.171 to 1.177. [less ▲]

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See detailIn vitro characterization of the Bacillus subtilis protein tyrosine phosphatase YwqE
Mijakovic, Ivan; Musumeci, Lucia ULg; Tautz, Lutz et al

in Journal of Bacteriology (2005), 187(10), 3384-90

Both gram-negative and gram-positive bacteria possess protein tyrosine phosphatases (PTPs) with a catalytic Cys residue. In addition, many gram-positive bacteria have acquired a new family of PTPs, whose ... [more ▼]

Both gram-negative and gram-positive bacteria possess protein tyrosine phosphatases (PTPs) with a catalytic Cys residue. In addition, many gram-positive bacteria have acquired a new family of PTPs, whose first characterized member was CpsB from Streptococcus pneumoniae. Bacillus subtilis contains one such CpsB-like PTP, YwqE, in addition to two class II Cys-based PTPs, YwlE and YfkJ. The substrates for both YwlE and YfkJ are presently unknown, while YwqE was shown to dephosphorylate two phosphotyrosine-containing proteins implicated in UDP-glucuronate biosynthesis, YwqD and YwqF. In this study, we characterize YwqE, compare the activities of the three B. subtilis PTPs (YwqE, YwlE, and YfkJ), and demonstrate that the two B. subtilis class II PTPs do not dephosphorylate the physiological substrates of YwqE. [less ▲]

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See detailIn Vitro Characterization Of The Homogalacturonan-Binding Domain Of The Wall-Associated Kinase Wak1 Using Site-Directed Mutagenesis
Decreux, Annabelle ULg; Thomas, Annick ULg; Spies, B. et al

in Phytochemistry (2006), 67(11), 1068-79

Wall-associated kinase 1--WAK1 is a transmembrane protein containing a cytoplasmic Ser/Thr kinase domain and an extracellular domain in contact with the pectin fraction of the plant cell wall in ... [more ▼]

Wall-associated kinase 1--WAK1 is a transmembrane protein containing a cytoplasmic Ser/Thr kinase domain and an extracellular domain in contact with the pectin fraction of the plant cell wall in Arabidopsis thaliana (L.) HEYNH. In a previous paper [Decreux, A., Messiaen, J., 2005. Wall-associated kinase WAK1 interacts with cell wall pectins in a calcium-induced conformation. Plant Cell Physiol. 46, 268-278], we showed that a recombinant peptide expressed in yeast corresponding to amino acids 67-254 of the extracellular domain of WAK1 specifically interacts with commercial non-methylesterified homogalacturonic acid, purified homogalacturonans from Arabidopsis and oligogalacturonides in a calcium-induced conformation. In this report, we used a receptor binding domain sequence-based prediction method to identify four putative binding sites in the extracellular domain of WAK1, in which cationic amino acids were selected for substitution by site-directed mutagenesis. Interaction studies between mutated forms of WAK1 and homogalacturonans allowed us to identify and confirm at least five specific amino acids involved in the interaction with homogalacturonan dimers and multimers. The presence of this homogalacturonan-binding domain within the extracellular domain of WAK1 is discussed in terms of cell wall architecture and signal transduction. [less ▲]

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See detailIN VITRO CLONAL PROPAGATION OF A PROMISING AGROFUEL PRODUCING-PLANT : JATROPHA CURCAS L.
Medza Mve, Samson Daudet ULg; Mergeai, Guy ULg; Baudoin, Jean-Pierre ULg et al

Poster (2009, June 13)

In the present investigation, in vitro clonal propagation of two-month-old Jatropha curcas L. was achieved employing nodal explants. Axillary shoot bud proliferation was best initiated on Murashige and ... [more ▼]

In the present investigation, in vitro clonal propagation of two-month-old Jatropha curcas L. was achieved employing nodal explants. Axillary shoot bud proliferation was best initiated on Murashige and Skoog’s (MS) basal medium supplemented with N6-benzyladenine (BA) and adenine sulphate. This medium allowed the production of 3.1 ± 0.5 shoots per nodal explant with 3.5 ± 0.8 cm average length after 3-4 weeks. [less ▲]

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See detailIn Vitro Comparison of the Antimycotic Activity of a Miconazole-Hp-Beta-Cyclodextrin Solution with a Miconazole Surfactant Solution
Piel, Géraldine ULg; Hayette, Marie-Pierre ULg; Pavoni, Ermanno et al

in Journal of Antimicrobial Chemotherapy (2001), 48(1), 83-7

The antimycotic activity of a new parenteral solution containing miconazole was compared with that of a marketed solution (Daktarin IV solution). This solution has been withdrawn from the Belgian market ... [more ▼]

The antimycotic activity of a new parenteral solution containing miconazole was compared with that of a marketed solution (Daktarin IV solution). This solution has been withdrawn from the Belgian market, probably because of toxic effects related to the presence of polyoxyl 35 castor oil. We propose a new formulation containing miconazole (10 mg/mL) (like the marketed solution), in combination with hydroxypropyl-beta-cyclodextrin and lactic acid. The MICs of these two solutions were determined by a broth microdilution method (based on NCCLS guidelines) for 67 yeasts and 50 filamentous fungi isolates. This study shows that the MICs obtained with these two solutions are not significantly different. [less ▲]

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See detailIn vitro culture of immature embryos of Phaseolus polyanthus Greenm. and Phaseolus vulgaris L.
Toussaint, André ULg; Clément, F.; Mergeai, Guy ULg et al

in Annual Report of the Bean Improvment Cooperative (2002), 45

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See detailIn vitro culture of seal muscle-derived satellite cells
Freichels, Astrid ULg; Baise, Etienne ULg; Jauniaux, Thierry ULg et al

Poster (2014, April)

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See detailIn vitro culture of the parasitic wasp Aphidius ervi: Sweet dream or Reality
Sabri, Ahmed ULg; Leroy, Pascal ULg; Haubruge, Eric ULg et al

Poster (2011, March 03)

Aphidius ervi is an entomophagous parasitoid and represents an effective parasitoid of several aphid species that cause great damages in agriculture. Several investigations, for its in vitro mass ... [more ▼]

Aphidius ervi is an entomophagous parasitoid and represents an effective parasitoid of several aphid species that cause great damages in agriculture. Several investigations, for its in vitro mass production, have achieved a limited success and suggest that in vitro culture of this valuable biological control agent is rather closer to a dream than reality. Our work provides a chronological study of A. ervi development, from the oviposition until hatching of the first instar larva, in the body of the pea aphid Acyrthosiphon pisum. We show that this parasitoid has some characteristics in its embryological development that are rather complex and different from most other reported insects, which can be phylogenetically very close. Some of these characteristics concern extraembryonic membranes and could be among the causes of the limited success achieved in the in vitro culture. [less ▲]

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See detailIn vitro cultures for producing pathogen-free plants and selecting disease resistant genotypes.
Semal, J.; Kummert, J.; Lepoivre, Philippe ULg et al

in Bulletin des Recherches Agronomiques de Gembloux (1988), 23(3),

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See detailIn Vitro Cytotoxic Activity of Two Potential Anticancer Drugs Isolated from Strychnos: Strychnopentamine and Usambarensine
Bonjean, K. A.; De Pauw-Gillet, Marie-Claire ULg; Quetin-Leclercq, J. et al

in Anticancer Research (1996), 16(3A, May-Jun), 1129-37

The cytotoxicity and the selective antiprotozoal activity of some Strychnos alkaloids, namely strychnopentamine (SP) and usambarensine (US) (7) led us to analyze and compare their effects with emetine (EM ... [more ▼]

The cytotoxicity and the selective antiprotozoal activity of some Strychnos alkaloids, namely strychnopentamine (SP) and usambarensine (US) (7) led us to analyze and compare their effects with emetine (EM) by using mouse B16 melanoma cells cultivated in vitro. We observed by cytological analysis and proliferation rate studies that these substances induce analogous cytotoxic effects in B16 cells, but at different concentrations i.e. formation of lamellar bodies in the cytoplasm, the which contain pre-melanosomes in the case of SP and US, vacuoles and blebs. At concentrations near their respective IC50, SP and US, but not EM, decreased colony formation. We showed by incorporation of labelled precursors that SP and US first inhibit RNA synthesis while EM initially acts on protein synthesis. These alkaloids increased melanin synthesis. Furthermore, only EM and SP caused hemolysis of sheep red blood corpuscles. This could explain why the rate of antiplasmodial activity is higher for SP and EM. [less ▲]

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See detailIn vitro cytotoxicity of some medicinal plants from Georgian amaryllidaceae
Jokhadze, Malkhaz; Eristavi, Lina; Kutchukhidze, Jumber et al

in Phytotherapy Research (2007), 21(7), 622-624

Using an ethnomedical data approach, some Georgian plants, which are used in Georgian traditional medicine for cancer or non-cancer diseases, were collected and evaluated for cytotoxic activity. The ... [more ▼]

Using an ethnomedical data approach, some Georgian plants, which are used in Georgian traditional medicine for cancer or non-cancer diseases, were collected and evaluated for cytotoxic activity. The cytotoxic effect of the methanol extracts of species from the genera Galanthus and Leucojum was evaluated in vitro on three human cell lines (Hela, ephitheloid cervix carcinoma; HCT-116, colon carcinoma; HL-60, acute myeloid leukaemia). Cell type cytotoxic specificity was observed for some extracts. Overall, the HCT-116 cells were much more sensitive to most of the extracts than were the other cell lines. Plants that showed pronounced cytotoxic activity will be further evaluated for the possible isolation of active antitumour compounds. Copyright (C) 2007 John Wiley [less ▲]

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See detailIn vitro degradation of the four isomers of Soman in human serum
De Bisschop, H. C.; Mainil, Jacques ULg; Willems, J. L.

in Biochemical Pharmacology (1985), 34

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See detailIn Vitro Detection And Characterization Of Bacteriocin-Like Inhibitory Activity Of Lactic Acid Bacteria (Lab) Isolated From Senegalese Local Food Products
Diop, Mb.; Dubois Dauphin, Robin ULg; Dortu, C. et al

in African Journal of Microbiology Research [=AJMR] (2008), 2(8),

The prevalence of lactic acid bacteria (LAB) in Senegalese local food products was determined to be 109 CFU/g in millet flour and milk products, and 103 CFU/g in seafood products. These food products are ... [more ▼]

The prevalence of lactic acid bacteria (LAB) in Senegalese local food products was determined to be 109 CFU/g in millet flour and milk products, and 103 CFU/g in seafood products. These food products are generally preserved by spontaneous fermentation (without addition of starters). Of 220 lactic acid bacteria strains randomly selected from such products, 12 isolates capable of producing bacteriocin-like substances (bac+) were detected. Based on the use of API 50 CH test kits and 16S rDNA sequencing, 11 isolates were characterized as Lactococcus lactis subsp. lactis strains and one as an Enterococcus faecium strain. Nisin- and enterocin B-encoding genes were respectively identified in the bac+ lactococcal strains and the E. faecium strain. Since the bac+ Lc. lactis strains were isolated from different products, it suggests a high potential of growth by these strains in variable ecological environments. Expression of the nisin gene was indicated for one of the lactococcal strains, designated Lc. lactis subsp. lactis CWBI-B1410, which showed the highest in vitro antibacterial activity. An antibacterial preparation prepared from the CWBI-B1410 strain showed many similarities with nisin with regards to its inhibitory effects, heat resistance, protease sensitivity profile, as well as retention time of the antibacterial substances on a C18 column. These results suggest that a nisin-like substance is produced by the CWBI-B1410 strain. This strain has been selected for application as an additional barrier to supplementation with sodium chloride as a means to improve the bacterial quality of fish commodities in Senegal. [less ▲]

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See detailIn vitro determination of prebiotic potential of sugar beet pulp extracted candidates and influence of production method
François, Emmanuelle ULg; Goffin, Dorothée ULg; combo, Agnan Marie-Michelle et al

Poster (2012, June 12)

Various prebiotic hydrolysates were produced starting from sugar beet pulp (SBP) using alternative solution to traditional acidic hydolysis. SBP pectin was first extracted by acid extraction and ethanol ... [more ▼]

Various prebiotic hydrolysates were produced starting from sugar beet pulp (SBP) using alternative solution to traditional acidic hydolysis. SBP pectin was first extracted by acid extraction and ethanol precipitation. Then two technics are used to produced pectic oligosaccharides (POS) : enzymatic hydrolysis (EnzPOS : Rapidase Smart (DSM) ; 50°C ; pH 5) and microwaves-assisted hydrolysis (MW1POS : 104°C/10min/pH 7; MW2POS : 110°C/15min/pH 7). Structural characterization of fractions gave rise to different structural profiles between the four products. Moreover, fermentation parameters obtained in-vitro (A, B, Rmax and Tmax) were also characteristic of the fractions and in favor of a tight relationship between POS structure and POS function. Finally, to avoid solvent use, POS production could be envisaged directly on SBP. Avoiding the step of acidic extraction will allow to meet the green chemistry principles. [less ▲]

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See detailIn vitro drug sensitivity and clinical aspects of Plasmodium falciparum malaria in African children
Carme, Bernard; Gay, Frederic; Ndounga, Mathieu et al

in Tropical Medicine and Parasitology : Official Organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft für Technische Zusammenarbeit (1995), 46(4), 270-274

In vitro Plasmodium falciparum drug sensitivity was investigated in 115 brazzavillians children, between 1 year and 10 years of age. On the basis of clinical aspects, four groups were constituted: Group 1 ... [more ▼]

In vitro Plasmodium falciparum drug sensitivity was investigated in 115 brazzavillians children, between 1 year and 10 years of age. On the basis of clinical aspects, four groups were constituted: Group 1: 39 asymptomatic school children, Group 2: 16 children with uncomplicated malaria, Group 3: 40 with severe but not pernicious malaria and Group 4: 20 with pernicious malaria. The drugs tested were chloroquine (CQ), quinine (QN) and mefloquine (MQ). The sensitivity level was assessed by a 48-hour in vitro maturation test involving the uptake of tritiated hypoxanthine, the initial blood level of parasite being > or = 0.1% in all cases. For QN and MQ, the median IC50 values showed no significant difference related to clinical status, age or parasitaemia levels. For CQ, the proportion of resistant strains and the 50 inhibitory concentration (IC50) values were greater in the cases of children hospitalised for malaria but there were no differences related to clinical severity of these hospitalised children nor, within each group, to the age or parasitaemia levels. The percentage of subjects with an IC50 value greater than the 90 percentile of the IC50 of the asymptomatic group, which we propose as the severity index related to chemoresistance, was 15% for uncomplicated malaria, 38% for severe but non-pernicious forms and 35% for pernicious malaria. The IC50 for QN was significantly higher in CQ-resistant strains and there was a positive correlation for CQ vs QN and for QN vs MQ. [less ▲]

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